SUMMARY OF THE INVENTION <EOH>The present invention is based in part on the identification of amino acid sequences of human Ras-like protein polypeptides and proteins that are related to the Ras-like GTPase protein subfamily, as well as allelic variants and other mammalian orthologs thereof.","These unique peptide sequences, and nucleic acid sequences that encode these peptides, can be used as models for the development of human therapeutic targets, aid in the identification of therapeutic proteins, and serve as targets for the development of human therapeutic agents that modulate Ras-like protein activity in cells and tissues that express the Ras-like protein.","Experimental data as provided in FIG.","1 indicates expression in bone marrow, stem cells, and leukocytes."],["FIELD OF THE INVENTION The present invention is in the field of Ras-like proteins that are related to the Ras-like GTPase subfamily, recombinant DNA molecules and protein production.","The present invention specifically provides novel Ras-like protein polypeptides and proteins and nucleic acid molecules encoding such peptide and protein molecules, all of which are useful in the development of human therapeutics and diagnostic compositions and methods.","BACKGROUND OF THE INVENTION Ras-like proteins, particularly members of the Ras-like GTPase subfamilies, are a major target for drug action and development.","Accordingly, it is valuable to the field of pharmaceutical development to identify and characterize previously unknown members of this subfamily of Ras-like proteins.","The present invention advances the state of the art by providing a previously unidentified human Ras-like proteins that have homology to members of the Ras-like GTPase subfamilies.","Ras Protein Ras proteins are small regulatory GTP-binding proteins, or small G proteins, which belong to the Ras protein superfamily.","They are monomeric GTPases, but their GTPase activity is very slow (less than one GTP molecule per minute).","Ras proteins are key relays in the signal-transducing cascade induced by the binding of a ligand to specific receptors such as receptor tyrosine kinases (RTKs), since they trigger the MAP kinase cascade.","The ligand can be a growth factor (epidermal growth factor (EGF), platelet-derived growth factor (PDGF), insulin, an interleukin (IL), granulocyte colony-stimulating factor (G-CSF), granulocyte/macrophage colony-stimulating factor (GM-CSF).","Ras proteins contain sequences highly conserved during evolution.","Their tertiary structure includes ten loops connecting six strands of beta-sheet and five alpha helices.","In mammalians, there are four Ras proteins, which are encoded by Ha-ras, N-ras, Ki-rasA and Ki-rasB genes.","They are composed of about 170 residues and have a relative molecular mass of 21 kD.","Ras proteins contain covalently attached modified lipids allowing these proteins to bind to the plasma membrane.","Ha-Ras has a C-terminal farnesyl group, a C-terminal palmitoyl group and a N-terminal myristoyl group.","In Ki-Ras(B), a C-terminal polylysine domain replaces the palmitoyl group.","Ras proteins alternate between an inactive form bound to GDP and an active form bound to GTP.","Their activation results from reactions induced by a guanine nucleotide-exchange factor (GEF).","Their inactivation results from reactions catalyzed by a GTPase-activating protein (GAP).","When a Ras protein is activated by a GEF such as a Sos protein, the N-terminal region of a serine/threonine kinase, called “Raf protein”, can bind to Ras protein.","The C-terminal region of the activated Raf thus formed binds to another protein, MEK, and phosphorylates it on both specific tyrosine and serine residues.","Active MEK phosphorylates and activates, in turn, a MAP kinase (ERK1 or ERK2), which is also a serine/threonine kinase.","This phosphorylation occurs on both specific tyrosine and threonine residues of MAP kinase.","MAP kinase phosphorylates many different proteins, especially nuclear transcription factors (TFs) that regulate expression of many genes during cell proliferation and differentiation.","Recent researches suggest that, in mammalians, phosphatidyl inositol 3′-kinase (PI3-kinase) might be a target of Ras protein, instead of Raf protein.","In certain mutations, the translation of ras genes may produce oncogenic Ras proteins.","Ras-Like Protein Guanine nucleotide-binding proteins (GTP-binding proteins, or G proteins) participate in a wide range of regulatory functions including metabolism, growth, differentiation, signal transduction, cytoskeletal organization, and intracellular vesicle transport and secretion.","These proteins control diverse sets of regulatory pathways in response to hormones, growth factors, neuromodulators, or other signaling molecules.","When these molecules bind to transmembrane receptors, signals are propagated to effector molecules by intracellular signal transducing proteins.","Many of these signal-transducing proteins are members of the Ras superfamily.","The Ras superfamily is a class of low molecular weight (LMW) GTP-binding proteins that consist of 21-30 kDa polypeptides.","These proteins regulate cell growth, cell cycle control, protein secretion, and intracellular vesicle interaction.","In particular, the LMW GTP-binding proteins activate cellular proteins by transducing mitogenic signals involved in various cell functions in response to extracellular signals from receptors (Tavitian, A.","(1995) C. R. Seances Soc.","Biol.","Fil.","189:7-12).","During this process, the hydrolysis of GTP acts as an energy source as well as an on-off switch for the GTPase activity of the LMW GTP-binding proteins.","The Ras superfamily is comprised of five subfamilies: Ras, Rho, Ran, Rab, and ADP-ribosylation factor (ARF).","Specifically, Ras genes are essential in the control of cell proliferation.","Mutations in Ras genes have been associated with cancer.","Rho proteins control signal transduction in the process of linking receptors of growth factors to actin polymerization that is necessary for cell division.","Rab proteins control the translocation of vesicles to and from membranes for protein localization, protein processing, and secretion.","Ran proteins are localized to the cell nucleus and play a key role in nuclear protein import, control of DNA synthesis, and cell-cycle progression.","ARF and ARF-like proteins participate in a wide variety of cellular functions including vesicle trafficking, exocrine secretion, regulation of phospholipase activity, and endocytosis.","Despite their sequence variations, all five subfamilies of the Ras superfamily share conserved structural features.","Four conserved sequence regions (motifs I-IV) have been studied in the LMW GTP-binding proteins.","Motif I is the most variable but has the conserved sequence, GXXXXGK.","The lysine residue is essential in interacting with the beta.- and .gamma.-phosphates of GTP.","Motif II, III, and IV contain highly conserved sequences of DTAGQ, NKXD, and EXSAX, respectively.","Specifically, Motif II regulates the binding of gamma-phosphate of GTP; Motif III regulates the binding of GTP; and Motif IV regulates the guanine base of GTP.","Most of the membrane-bound LMW GTP-binding proteins generally require a carboxy terminal isoprenyl group for membrane association and biological activity.","The isoprenyl group is added posttranslationally through recognition of a terminal cysteine residue alone or a terminal cysteine-aliphatic amino acid-aliphatic amino acid-any amino acid (CAAX) motif.","Additional membrane-binding energy is often provided by either internal palmitoylation or a carboxy terminal cluster of basic amino acids.","The LMW GTP-binding proteins also have a variable effector region, located between motifs I and II, which is characterized as the interaction site for guanine nucleotide exchange factors (GEFs) or GTPase-activating proteins (GAPs).","GEFs induce the release of GDP from the active form of the G protein, whereas GAPs interact with the inactive form by stimulating the GTPase activity of the G protein.","The ARF subfamily has at least 15 distinct members encompassing both ARF and ARF-like proteins.","ARF proteins identified to date exhibit high structural similarity and ADP-ribosylation enhancing activity.","In contrast, several ARF-like proteins lack ADP-ribosylation enhancing activity and bind GTP differently.","An example of ARF-like proteins is a rat protein, ARL184.ARL184 has been shown to have a molecular weight of 22 kDa and four functional GTP-binding sites (Icard-Liepkalns, C. et al., (1997) Eur.","J. Biochem.","246: 388-393).","ARL184 is active in both the cytosol and the Golgi apparatus and is closely associated with acetylcholine release, suggesting that ARL184 is a potential regulatory protein associated with Ca.sup.2+-dependent release of acetylcholine.","A number of Rho GTP-binding proteins have been identified in plasma membrane and cytoplasm.","These include RhoA, B and C, and D, rhoG, rac 1 and 2, G25K-A and B, and TC10 (Hall, A. et al.","(1993) Philos.","Trans.","R. Soc.","Lond.","(Biol.)","340:267-271).","All Rho proteins have a CAAX motif that binds a prenyl group and either a palmitoylation site or a basic amino acid-rich region, suggesting their role in membrane-associated functions.","In particular, RhoD is a protein that functions in early endosome motility and distribution by inducing rearrangement of actin cytoskeleton and cell surface (Murphy, C. et al.","(1996) Nature 384:427-432).","During cell adhesion, the Rho proteins are essential for triggering focal complex assembly and integrin-dependent signal transduction (Hotchin, N. A. and Hall, A.","(1995) J.","Cell Biol.","131:1857-1865).","The Ras subfamily proteins already indicated supra are essential in transducing signals from receptor tyrosine kinases (RTKs) to a series of serine/threonine kinases which control cell growth and differentiation.","Mutant Ras proteins, which bind but cannot hydrolyze GTP, are permanently activated and cause continuous cell proliferation or cancer.","TC21, a Ras-like protein, is found to be highly expressed in a human teratocarcinoma cell line (Drivas, G. T. et al.","(1990) Mol.","Cell.","Biol.","10: 1793-1798).","Rin and Rit are characterized as membrane-binding, Ras-like proteins without the lipid-binding CAAX motif and carboxy terminal cysteine (Lee, C.-H. J. et al.","(1996) J. Neurosci.","16: 6784-6794).","Further, Rin is shown to localize in neurons and have calcium-dependant calmodulin-binding activity.","Ras-Like GTPase Proteins The novel human protein, and encoding gene, provided by the present invention is related to the family of Ras-like GTPase proteins (also referred to as Ras-like GTP-binding proteins), which includes Rab proteins.","The protein of the present invention is similar to Rab8 and low molecular weight (LMW) GTP-binding proteins isolated from an electrode lobe library of the marine ray Discopyge ommata (Ngsee et al., J. Biol.","Chem.","266 (4), 2675-2680 (1991)), some of which were determined to be homologs of the rab1, ra1, Krev, and rho LMW GTP-binding proteins.","These proteins were localized to cholinergic synaptic vesicles and at least two of these proteins, oral and o-rho, were localized to the pre-synaptic terminals (Ngsee et al., J. Biol.","Chem.","266 (4), 2675-2680 (1991)).","Rab proteins are important for regulating the targeting and fusion of membranous vesicles during organelle assembly and transport.","Rab proteins undergo controlled exchange of GTP for GDP, and they hydrolyze GTP in a reaction that may regulate the timing and unidirectional nature of these assemblies.","Generally, known Rab proteins terminate in sequences such as cys-X-cys (e.g., RAB3A), cys-cys (e.g., RAB1A), or a similar sequence, and generally all are geranylgeranylated.","Rab GTP-binding proteins are similar to YPT1/SEC4 in Saccharomyces cerevisiae, which are critical for transport along the exocytic route (Chavrier et al., Mol Cell Biol 1990 December;10(12):6578-85).","Different Rab proteins are presumed to control different steps in membrane traffic, leading to a high level of diversity and complexity within the Rab family (Chavrier et al., Mol Cell Biol 1990 December;10(12):6578-85).","The Rab1 gene maps in close viscinity to the ‘wobbler’ spinal muscular atrophy gene.","Rab1 and Rab2 from the snail Lymnaea stagnalis share a very high degree (95-97%) of sequence identity with mammalian Rab1 and Rab2 over the first 178-191 N-terminal amino acids; however, the C-terminal region is almost completely divergent, except for the final 24 amino acids at the extreme ends.","Rab1 was found to be highly expressed in the albumin gland of Lymnaea stagnalis, suggesting an important role in that gland (Agterberg et al., Eur.","J. Biochem.","217 (1), 241-246 (1993)).","The tethering factor p115 is a RAB1 effector that binds directly to activated RAB1.It is thought that RAB1-regulated assembly of functional effector-SNARE complexes serves as a conserved molecular mechanism for regulating recognition between different subcellular compartments such as endoplasmic reticulum and Golgi apparatus (Allan et al., Science 289: 444-448, 2000).","GTPases play important roles in a wide variety of cell functions such as signal transduction, cytoskeletal organization, and membrane trafficking.","Rab GTPases are particularly important for regulating cellular membrane dynamics by modulating the activity of effector proteins that then regulate vesicle trafficking.","The Rab8 GTPase plays important roles in Golgi to plasma membrane vesicle trafficking.","Studies have suggested that Rab37 plays an important role in mast cell degranulation.","Thus, novel human Rab GTPases may be valuable as potential therapeutic targets for the development of allergy treatments (Masuda et al., FEBS Lett 2000 Mar.","17;470).","Rab5 may act, together with Rab3A, to regulate synaptic vesicle membrane flow within nerve terminals, thereby regulating neurotransmitter release.","Rab15 and Rab3A are low molecular weight GTP-binding proteins.","Rab proteins are generally comprised of four conserved structural domains necessary for GTP binding, as well as additional domains for membrane localization and effector protein interactions.","Rab15 is expressed primarily in neural tissues such as the brain and is localized to synaptic vesicles (Elferink et al., J. Biol.","Chem.","267 (9), 5768-5775 (1992)).","For a further review of Rab proteins, see Wedemeyer et al., Genomics 32: 447-454, 1996 and Zahraoui et al., J. Biol.","Chem.","264: 12394-12401, 1989.Due to their importance in human physiology, particularly in regulating membrane trafficking, novel human Ras-like GTPase proteins/genes, such as provided by the present invention, are valuable as potential targets for the development of therapeutics to treat a wide variety of diseases/disorders caused or influenced by defects in membrane trafficking.","Furthermore, SNPs in Ras-like GTPase genes are valuable markers for the diagnosis, prognosis, prevention, and/or treatment of such diseases/disorders.","Using the information provided by the present invention, reagents such as probes/primers for detecting the SNPs or the expression of the protein/gene provided herein may be readily developed and, if desired, incorporated into kit formats such as nucleic acid arrays, primer extension reactions coupled with mass spec detection (for SNP detection), or TaqMan PCR assays (Applied Biosystems, Foster City, Calif.).","The discovery of new human Ras-like proteins and the polynucleotides that encode them satisfies a need in the art by providing new compositions that are useful in the diagnosis, prevention, and treatment of inflammation and disorders associated with cell proliferation and apoptosis.","SUMMARY OF THE INVENTION The present invention is based in part on the identification of amino acid sequences of human Ras-like protein polypeptides and proteins that are related to the Ras-like GTPase protein subfamily, as well as allelic variants and other mammalian orthologs thereof.","These unique peptide sequences, and nucleic acid sequences that encode these peptides, can be used as models for the development of human therapeutic targets, aid in the identification of therapeutic proteins, and serve as targets for the development of human therapeutic agents that modulate Ras-like protein activity in cells and tissues that express the Ras-like protein.","Experimental data as provided in FIG.","1 indicates expression in bone marrow, stem cells, and leukocytes.","DESCRIPTION OF THE FIGURE SHEETS FIG.","1 provides the nucleotide sequence of a cDNA molecule that encodes the Ras-like protein of the present invention.","(SEQ ID NO:1) In addition, structure and functional information is provided, such as ATG start, stop and tissue distribution, where available, that allows one to readily determine specific uses of inventions based on this molecular sequence.","Experimental data as provided in FIG.","1 indicates expression in bone marrow, stem cells, and leukocytes.","FIG.","2 provides the predicted amino acid sequence of the Ras-like protein of the present invention.","(SEQ ID NO:2) In addition structure and functional information such as protein family, function, and modification sites is provided where available, allowing one to readily determine specific uses of inventions based on this molecular sequence.","FIG.","3 provides genomic sequences that span the gene encoding the Ras-like protein of the present invention.","(SEQ ID NO:3) In addition structure and functional information, such as intron/exon structure, promoter location, etc., is provided where available, allowing one to readily determine specific uses of inventions based on this molecular sequence.","As illustrated in FIG.","3, SNPs were identified at 37 different nucleotide positions.","DETAILED DESCRIPTION OF THE INVENTION General Description The present invention is based on the sequencing of the human genome.","During the sequencing and assembly of the human genome, analysis of the sequence information revealed previously unidentified fragments of the human genome that encode peptides that share structural and/or sequence homology to protein/peptide/domains identified and characterized within the art as being a Ras-like protein or part of a Ras-like protein and are related to the Ras-like GTPase subfamily.","Utilizing these sequences, additional genomic sequences were assembled and transcript and/or cDNA sequences were isolated and characterized.","Based on this analysis, the present invention provides amino acid sequences of human Ras-like protein polypeptides that are related to the Ras-like GTPase subfamily, nucleic acid sequences in the form of transcript sequences, cDNA sequences and/or genomic sequences that encode these Ras-like protein polypeptide, nucleic acid variation (allelic information), tissue distribution of expression, and information about the closest art known protein/peptide/domain that has structural or sequence homology to the Ras-like protein of the present invention.","In addition to being previously unknown, the peptides that are provided in the present invention are selected based on their ability to be used for the development of commercially important products and services.","Specifically, the present peptides are selected based on homology and/or structural relatedness to known Ras-like proteins of the Ras-like GTPase subfamily and the expression pattern observed.","Experimental data as provided in FIG.","1 indicates expression in bone marrow, stem cells, and leukocytes.","The art has clearly established the commercial importance of members of this family of proteins and proteins that have expression patterns similar to that of the present gene.","Some of the more specific features of the peptides of the present invention, and the uses thereof, are described herein, particularly in the Background of the Invention and in the annotation provided in the Figures, and/or are known within the art for each of the known Ras-like GTPase family or subfamily of Ras-like proteins.","Specific Embodiments Peptide Molecules The present invention provides nucleic acid sequences that encode protein molecules that have been identified as being members of the Ras-like protein family and are related to the Ras-like GTPase subfamily (protein sequences are provided in FIG.","2, transcript/cDNA sequences are provided in FIG.","1 and genomic sequences are provided in FIG.","3).","The peptide sequences provided in FIG.","2, as well as the obvious variants described herein, particularly allelic variants as identified herein and using the information in FIG.","3, will be referred herein as the Ras-like proteins or peptides of the present invention, Ras-like proteins or peptides, or peptides/proteins of the present invention.","The present invention provides isolated peptide and protein molecules that consist of, consist essentially of, or comprise the amino acid sequences of the Ras-like protein polypeptide disclosed in the FIG.","2, (encoded by the nucleic acid molecule shown in FIG.","1, transcript/cDNA or FIG.","3, genomic sequence), as well as all obvious variants of these peptides that are within the art to make and use.","Some of these variants are described in detail below.","As used herein, a peptide is said to be “isolated” or “purified” when it is substantially free of cellular material or free of chemical precursors or other chemicals.","The peptides of the present invention can be purified to homogeneity or other degrees of purity.","The level of purification will be based on the intended use.","The critical feature is that the preparation allows for the desired function of the peptide, even if in the presence of considerable amounts of other components.","In some uses, “substantially free of cellular material” includes preparations of the peptide having less than about 30% (by dry weight) other proteins (i.e., contaminating protein), less than about 20% other proteins, less than about 10% other proteins, or less than about 5% other proteins.","When the peptide is recombinantly produced, it can also be substantially free of culture medium, i.e., culture medium represents less than about 20% of the volume of the protein preparation.","The language “substantially free of chemical precursors or other chemicals” includes preparations of the peptide in which it is separated from chemical precursors or other chemicals that are involved in its synthesis.","In one embodiment, the language “substantially free of chemical precursors or other chemicals” includes preparations of the Ras-like protein polypeptide having less than about 30% (by dry weight) chemical precursors or other chemicals, less than about 20% chemical precursors or other chemicals, less than about 10% chemical precursors or other chemicals, or less than about 5% chemical precursors or other chemicals.","The isolated Ras-like protein polypeptide can be purified from cells that naturally express it, purified from cells that have been altered to express it (recombinant), or synthesized using known protein synthesis methods.","Experimental data as provided in FIG.","1 indicates expression in bone marrow, stem cells, and leukocytes.","For example, a nucleic acid molecule encoding the Ras-like protein polypeptide is cloned into an expression vector, the expression vector introduced into a host cell and the protein expressed in the host cell.","The protein can then be isolated from the cells by an appropriate purification scheme using standard protein purification techniques.","Many of these techniques are described in detail below.","Accordingly, the present invention provides proteins that consist of the amino acid sequences provided in FIG.","2 (SEQ ID NO:2), for example, proteins encoded by the transcript/cDNA nucleic acid sequences shown in FIG.","1 (SEQ ID NO:1) and the genomic sequences provided in FIG.","3 (SEQ ID NO:3).","The amino acid sequence of such a protein is provided in FIG.","2.A protein consists of an amino acid sequence when the amino acid sequence is the final amino acid sequence of the protein.","The present invention further provides proteins that consist essentially of the amino acid sequences provided in FIG.","2 (SEQ ID NO:2), for example, proteins encoded by the transcript/cDNA nucleic acid sequences shown in FIG.","1 (SEQ ID NO:1) and the genomic sequences provided in FIG.","3 (SEQ ID NO:3).","A protein consists essentially of an amino acid sequence when such an amino acid sequence is present with only a few additional amino acid residues, for example from about 1 to about 100 or so additional residues, typically from 1 to about 20 additional residues in the final protein.","The present invention further provides proteins that comprise the amino acid sequences provided in FIG.","2 (SEQ ID NO:2), for example, proteins encoded by the transcript/cDNA nucleic acid sequences shown in FIG.","1 (SEQ ID NO:1) and the genomic sequences provided in FIG.","3 (SEQ ID NO:3).","A protein comprises an amino acid sequence when the amino acid sequence is at least part of the final amino acid sequence of the protein.","In such a fashion, the protein can be only the peptide or have additional amino acid molecules, such as amino acid residues (contiguous encoded sequence) that are naturally associated with it or heterologous amino acid residues/peptide sequences.","Such a protein can have a few additional amino acid residues or can comprise several hundred or more additional amino acids.","The preferred classes of proteins that are comprised of the Ras-like protein polypeptide of the present invention are the naturally occurring mature proteins.","A brief description of how various types of these proteins can be made/isolated is provided below.","The Ras-like protein polypeptides of the present invention can be attached to heterologous sequences to form chimeric or fusion proteins.","Such chimeric and fusion proteins comprise a Ras-like protein polypeptide operatively linked to a heterologous protein having an amino acid sequence not substantially homologous to the Ras-like protein polypeptide.","“Operatively linked” indicates that the Ras-like protein polypeptide and the heterologous protein are fused in-frame.","The heterologous protein can be fused to the N-terminus or C-terminus of the Ras-like protein polypeptide.","In some uses, the fusion protein does not affect the activity of the Ras-like protein polypeptide per se.","For example, the fusion protein can include, but is not limited to, enzymatic fusion proteins, for example beta-galactosidase fusions, yeast two-hybrid GAL fusions, poly-His fusions, MYC-tagged, HI-tagged and Ig fusions.","Such fusion proteins, particularly poly-His fusions, can facilitate the purification of recombinant Ras-like protein polypeptide.","In certain host cells (e.g., mammalian host cells), expression and/or secretion of a protein can be increased by using a heterologous signal sequence.","A chimeric or fusion protein can be produced by standard recombinant DNA techniques.","For example, DNA fragments coding for the different protein sequences are ligated together in-frame in accordance with conventional techniques.","In another embodiment, the fusion gene can be synthesized by conventional techniques including automated DNA synthesizers.","Alternatively, PCR amplification of gene fragments can be carried out using anchor primers which give rise to complementary overhangs between two consecutive gene fragments which can subsequently be annealed and re-amplified to generate a chirneric gene sequence (see Ausubel et al., Current Protocols in Molecular Biology, 1992).","Moreover, many expression vectors are commercially available that already encode a fusion moiety (e.g., a GST protein).","A Ras-like protein polypeptide-encoding nucleic acid can be cloned into such an expression vector such that the fusion moiety is linked in-frame to the Ras-like protein polypeptide.","As mentioned above, the present invention also provides and enables obvious variants of the amino acid sequence of the peptides of the present invention, such as naturally occurring mature forms of the peptide, allelic/sequence variants of the peptides, non-naturally occurring recombinantly derived variants of the peptides, and orthologs and paralogs of the peptides.","Such variants can readily be generated using art know techniques in the fields of recombinant nucleic acid technology and protein biochemistry.","It is understood, however, that variants exclude any amino acid sequences disclosed prior to the invention.","Such variants can readily be identified/made using molecular techniques and the sequence information disclosed herein.","Further, such variants can readily be distinguished from other peptides based on sequence and/or structural homology to the Ras-like protein polypeptides of the present invention.","The degree of homology/identity present will be based primarily on whether the peptide is a functional variant or non-functional variant, the amount of divergence present in the paralog family, and the evolutionary distance between the orthologs.","To determine the percent identity of two amino acid sequences or two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second amino acid or nucleic acid sequence for optimal alignment and non-homologous sequences can be disregarded for comparison purposes).","In a preferred embodiment, the length of a reference sequence aligned for comparison purposes is at least 30%, 40%, 50%, 60%, 70%, 80%, or 90% or more of the length of the reference sequence.","The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared.","When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position (as used herein amino acid or nucleic acid “identity” is equivalent to amino acid or nucleic acid “homology”).","The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which need to be introduced for optimal alignment of the two sequences.","The comparison of sequences and determination of percent identity and similarity between two sequences can be accomplished using a mathematical algorithm.","(Computational Molecular Biology, Lesk, A. M., ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, D. W., ed., Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part 1, Griffin, A. M., and Griffin, H. G., eds., Humana Press, New Jersey, 1994; Sequence Analysis in Molecular Biology, von Heinj e, G., Academic Press, 1987; and Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991).","In a preferred embodiment, the percent identity between two amino acid sequences is determined using the Needleman and Wunsch (J. Mol.","Biol.","(48):444-453 (1970)) algorithm which has been incorporated into the GAP program in the GCG software package (available at http://www.gcg.com), using either a Blossom 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6.In yet another preferred embodiment, the percent identity between two nucleotide sequences is determined using the GAP program in the GCG software package (Devereux, J., et al, Nucleic Acids Res.","12(1):387 (1984)) (available at http://www.gcg.com), using a NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 80 and a length weight of 1, 2, 3, 4, 5, or 6.In another embodiment, the percent identity between two amino acid or nucleotide sequences is determined using the algorithm of E. Meyers and W. Miller (CABIOS, 4:11-17 (1989)) which has been incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4.The nucleic acid and protein sequences of the present invention can further be used as a “query sequence” to perform a search against sequence databases to, for example, identify other family members or related sequences.","Such searches can be performed using the NBLAST and XBLAST programs (version 2.0) of Altschul, et al.","(J. Mol.","Biol.","215:403-10 (1990)).","BLAST nucleotide searches can be performed with the NBLAST program, score=100, word length=12 to obtain nucleotide sequences homologous to the nucleic acid molecules of the invention.","BLAST protein searches can be performed with the XBLAST program, score=50, word length=3, to obtain amino acid sequences homologous to the proteins of the invention.","To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al.","(Nucleic Acids Res.","25(17):3389-3402 (1997)).","When utilizing BLAST and gapped BLAST programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used.","See http://www.ncbi.nlm.nih.gov.","Full-length pre-processed forms, as well as mature processed forms, of proteins that comprise one of the peptides of the present invention can readily be identified as having complete sequence identity to one of the Ras-like protein polypeptides of the present invention as well as being encoded by the same genetic locus as the Ras-like protein polypeptide provided herein.","The gene encoding the novel Ras-like protein of the present invention is located on a genome component that has been mapped to human chromosome 6 (as indicated in FIG.","3), which is supported by multiple lines of evidence, such as STS and BAC map data.","Allelic variants of a Ras-like protein polypeptide can readily be identified as being a human protein having a high degree (significant) of sequence homology/identity to at least a portion of the Ras-like protein polypeptide as well as being encoded by the same genetic locus as the Ras-like protein polypeptide provided herein.","Genetic locus can readily be determined based on the genomic information provided in FIG.","3, such as the genomic sequence mapped to the reference human.","The gene encoding the novel Ras-like protein of the present invention is located on a genome component that has been mapped to human chromosome 6 (as indicated in FIG.","3), which is supported by multiple lines of evidence, such as STS and BAC map data.","As used herein, two proteins (or a region of the proteins) have significant homology when the amino acid sequences are typically at least about 70-80%, 80-90%, and more typically at least about 90-95% or more homologous.","A significantly homologous amino acid sequence, according to the present invention, will be encoded by a nucleic acid sequence that will hybridize to a Ras-like protein polypeptide encoding nucleic acid molecule under stringent conditions as more fully described below.","FIG.","3 provides information on SNPs that have been found in the gene encoding the Ras-like protein of the present invention.","SNPs were identified at 37 different nucleotide positions.","Some of these SNPs that are located outside the ORF and in introns may affect gene transcription.","Paralogs of a Ras-like protein polypeptide can readily be identified as having some degree of significant sequence homology/identity to at least a portion of the Ras-like protein polypeptide, as being encoded by a gene from humans, and as having similar activity or function.","Two proteins will typically be considered paralogs when the amino acid sequences are typically at least about 40-50%, 50-60%, and more typically at least about 60-70% or more homologous through a given region or domain.","Such paralogs will be encoded by a nucleic acid sequence that will hybridize to a Ras-like protein polypeptide encoding nucleic acid molecule under moderate to stringent conditions as more fully described below.","Orthologs of a Ras-like protein polypeptide can readily be identified as having some degree of significant sequence homology/identity to at least a portion of the Ras-like protein polypeptide as well as being encoded by a gene from another organism.","Preferred orthologs will be isolated from mammals, preferably primates, for the development of human therapeutic targets and agents.","Such orthologs will be encoded by a nucleic acid sequence that will hybridize to a Ras-like protein polypeptide encoding nucleic acid molecule under moderate to stringent conditions, as more fully described below, depending on the degree of relatedness of the two organisms yielding the proteins.","Non-naturally occurring variants of the Ras-like protein polypeptides of the present invention can readily be generated using recombinant techniques.","Such variants include, but are not limited to deletions, additions and substitutions in the amino acid sequence of the Ras-like protein polypeptide.","For example, one class of substitutions is conserved amino acid substitutions.","Such substitutions are those that substitute a given amino acid in a Ras-like protein polypeptide by another amino acid of like characteristics.","Typically seen as conservative substitutions are the replacements, one for another, among the aliphatic amino acids Ala, Val, Leu, and Ile; interchange of the hydroxyl residues Ser and Thr, exchange of the acidic residues Asp and Glu, substitution between the amide residues Asn and Gln, exchange of the basic residues Lys and Arg, replacements among the aromatic residues Phe, Tyr, and the like.","Guidance concerning which amino acid changes are likely to be phenotypically silent are found in Bowie et al., Science 247:1306-1310 (1990).","Variant Ras-like protein polypeptides can be fully functional or can lack function in one or more activities.","Fully functional variants typically contain only conservative variations or variations in non-critical residues or in non-critical regions.","Functional variants can also contain substitution of similar amino acids that result in no change or an insignificant change in function.","Alternatively, such substitutions may positively or negatively affect function to some degree.","Non-functional variants typically contain one or more non-conservative amino acid substitutions, deletions, insertions, inversions, or truncation or a substitution, insertion, inversion, or deletion in a critical residue or critical region.","Amino acids that are essential for function can be identified by methods known in the art, such as site-directed mutagenesis or alanine-scanning mutagenesis (Cunningham et al., Science 244:1081-1085 (1989)).","The latter procedure introduces single alanine mutations at every residue in the molecule.","The resulting mutant molecules are then tested for biological activity such as receptor binding or in vitro proliferative activity.","Sites that are critical for ligand-receptor binding can also be determined by structural analysis such as crystallography, nuclear magnetic resonance, or photoaffinity labeling (Smith et al., J. Mol.","Biol.","224:899-904 (1992); de Vos et al.","Science 255:306-312 (1992)).","The present invention further provides fragments of the Ras-like protein polypeptides, in addition to proteins and peptides that comprise and consist of such fragments.","Particularly those comprising the residues identified in FIG.","2.The fragments to which the invention pertains, however, are not to be construed as encompassing fragments that have been disclosed publicly prior to the present invention.","As used herein, a fragment comprises at least 8, 10, 12, 14, 16 or more contiguous amino acid residues from a Ras-like protein polypeptide.","Such fragments can be chosen based on the ability to retain one or more of the biological activities of the Ras-like protein polypeptide, or can be chosen for the ability to perform a function, e.g., act as an immunogen.","Particularly important fragments are biologically active fragments, peptides that are, for example about 8 or more amino acids in length.","Such fragments will typically comprise a domain or motif of the Ras-like protein polypeptide, e.g., active site.","Further, possible fragments include, but are not limited to, domain or motif containing fragments, soluble peptide fragments, and fragments containing immunogenic structures.","Predicted domains and functional sites are readily identifiable by computer programs well known and readily available to those of skill in the art (e.g., PROSITE, HMMer, eMOTIF, etc.).","The results of one such analysis are provided in FIG.","2.Polypeptides often contain amino acids other than the 20 amino acids commonly referred to as the 20 naturally occurring amino acids.","Further, many amino acids, including the terminal amino acids, may be modified by natural processes, such as processing and other post-translational modifications, or by chemical modification techniques well known in the art.","Common modifications that occur naturally in Ras-like protein polypeptides are described in basic texts, detailed monographs, and the research literature, and they are well known to those of skill in the art (some of these features are identified in FIG.","2).","Known modifications include, but are not limited to, acetylation, acylation, ADP-ribosylation, amidation, covalent attachment of flavin, covalent attachment of a heme moiety, covalent attachment of a nucleotide or nucleotide derivative, covalent attachment of a lipid or lipid derivative, covalent attachment of phosphotidylinositol, cross-linking, cyclization, disulfide bond formation, demethylation, formation of covalent crosslinks, formation of cystine, formation of pyroglutamate, formylation, gamma carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, myristoylation, oxidation, proteolytic processing, phosphorylation, prenylation, racemization, selenoylation, sulfation, transfer-RNA mediated addition of amino acids to proteins such as arginylation, and ubiquitination.","Such modifications are well known to those of skill in the art and have been described in great detail in the scientific literature.","Several particularly common modifications, glycosylation, lipid attachment, sulfation, gamma-carboxylation of glutamic acid residues, hydroxylation and ADP-ribosylation, for instance, are described in most basic texts, such as Proteins—Structure and Molecular Properties, 2nd Ed., T. E. Creighton, W. H. Freeman and Company, New York (1993).","Many detailed reviews are available on this subject, such as by Wold, F., Posttranslational Covalent Modification of proteins, B. C. Johnson, Ed., Academic Press, New York 1-12 (1983); Seifter et al.","(Meth.","Enzymol.","182: 626-646 (1990)) and Rattan et al.","(Ann.","N.Y. Acad.","Sci.","663:48-62 (1992)).","Accordingly, the Ras-like protein polypeptides of the present invention also encompass derivatives or analogs in which a substituted amino acid residue is not one encoded by the genetic code, in which a substituent group is included, in which the mature Ras-like protein polypeptide is fused with another compound, such as a compound to increase the half-life of the Ras-like protein polypeptide (for example, polyethylene glycol), or in which the additional amino acids are fused to the mature Ras-like protein polypeptide, such as a leader or secretory sequence or a sequence for purification of the mature Ras-like protein polypeptide, or a pro-protein sequence.","Protein/Peptide Uses The proteins of the present invention can be used in assays to determine the biological activity of the protein, including in a panel of multiple proteins for high-throughput screening; to raise antibodies or to elicit another immune response; as a reagent (including the labeled reagent) in assays designed to quantitatively determine levels of the protein (or its ligand or receptor) in biological fluids; and as markers for tissues in which the corresponding protein is preferentially expressed (either constitutively or at a particular stage of tissue differentiation or development or in a disease state).","Where the protein binds or potentially binds to another protein (such as, for example, in a receptor-ligand interaction), the protein can be used to identify the binding partner so as to develop a system to identify inhibitors of the binding interaction.","Any or all of these research utilities are capable of being developed into reagent grade or kit format for commercialization as research products.","Methods for performing the uses listed above are well known to those skilled in the art.","References disclosing such methods include “Molecular Cloning: A Laboratory Manual”, 2d ed., Cold Spring Harbor Laboratory Press, Sambrook, J., E. F. Fritsch and T. Maniatis eds., 1989, and “Methods in Enzymology: Guide to Molecular Cloning Techniques”, Academic Press, Berger, S. L. and A. R. Kimmel eds., 1987.The potential uses of the peptides of the present invention are based primarily on the source of the protein as well as the class/action of the protein.","For example, Ras-like proteins isolated from humans and their human/mammalian orthologs serve as targets for identifying agents for use in mammalian therapeutic applications, e.g.","a human drug, particularly in modulating a biological or pathological response in a cell or tissue that expresses the Ras-like protein.","Experimental data as provided in FIG.","1 indicates that Ras-like proteins of the present invention are expressed in bone marrow and stem cells (as indicated by virtual northern blot analysis), and leukocytes (as indicated by PCR-based tissue screening panels).","A large percentage of pharmaceutical agents are being developed that modulate the activity of Ras-like proteins, particularly members of the Ras-like GTPase subfamily (see Background of the Invention).","The structural and functional information provided in the Background and Figures provide specific and substantial uses for the molecules of the present invention, particularly in combination with the expression information provided in FIG.","1.Experimental data as provided in FIG.","1 indicates expression in bone marrow, stem cells, and leukocytes.","Such uses can readily be determined using the information provided herein, that which is known in the art, and routine experimentation.","The proteins of the present invention (including variants and fragments that may have been disclosed prior to the present invention) are useful for biological assays related to Ras-like proteins that are related to members of the Ras-like GTPase subfamily.","Such assays involve any of the known Ras-like protein functions or activities or properties useful for diagnosis and treatment of Ras-like protein-related conditions that are specific for the subfamily of Ras-like proteins that the one of the present invention belongs to, particularly in cells and tissues that express the Ras-like protein.","Experimental data as provided in FIG.","1 indicates that Ras-like proteins of the present invention are expressed in bone marrow and stem cells (as indicated by virtual northern blot analysis), and leukocytes (as indicated by PCR-based tissue screening panels).","The proteins of the present invention are also useful in drug screening assays, in cell-based or cell-free systems.","Cell-based systems can be native, i.e., cells that normally express the Ras-like protein, as a biopsy or expanded in cell culture.","Experimental data as provided in FIG.","1 indicates expression in bone marrow, stem cells, and leukocytes.","In an alternate embodiment, cell-based assays involve recombinant host cells expressing the Ras-like protein.","The polypeptides can be used to identify compounds that modulate Ras-like protein activity.","Both the Ras-like protein of the present invention and appropriate variants and fragments can be used in high-throughput screens to assay candidate compounds for the ability to bind to the Ras-like protein.","These compounds can be further screened against a functional Ras-like protein to determine the effect of the compound on the Ras-like protein activity.","Further, these compounds can be tested in animal or invertebrate systems to determine activity/effectiveness.","Compounds can be identified that activate (agonist) or inactivate (antagonist) the Ras-like protein to a desired degree.","Therefore, in one embodiment, Ras-like GTPase or a fragment or derivative thereof may be administered to a subject to prevent or treat a disorder associated with an increase in apoptosis.","Such disorders include, but are not limited to, AIDS and other infectious or genetic immunodeficiencies, neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, retinitis pigmentosa, and cerebellar degeneration, myelodysplastic syndromes such as aplastic anemia, ischemic injuries such as myocardial infarction, stroke, and reperfusion injury, toxin-induced diseases such as alcohol-induced liver damage, cirrhosis, and lathyrism, wasting diseases such as cachexia, viral infections such as those caused by hepatitis B and C, and osteoporosis.","In another embodiment, a pharmaceutical composition comprising Ras-like GTPase may be administered to a subject to prevent or treat a disorder associated with increased apoptosis including, but not limited to, those listed above.","In still another embodiment, an agonist which is specific for Ras-like GTPase may be administered to prevent or treat a disorder associated with increased apoptosis including, but not limited to, those listed above.","In a further embodiment, a vector capable of expressing Ras-like GTPase, or a fragment or a derivative thereof, may be used to prevent or treat a disorder associated with increased apoptosis including, but not limited to, those listed above.","In cancer, where Ras-like GTPase promotes cell proliferation, it is desirable to decrease its activity.","Therefore, in one embodiment, an antagonist of Ras-like GTPase may be administered to a subject to prevent or treat cancer including, but not limited to, adenocarcinoma, leukemia, lymphoma, melanoma, myeloma, sarcoma, and teratocarcinoma, and, in particular, cancers of the adrenal gland, bladder, bone, bone marrow, brain, breast, cervix, gall bladder, ganglia, gastrointestinal tract, heart, kidney, liver, lung, muscle, ovary, pancreas, parathyroid, penis, prostate, salivary glands, skin, spleen, testis, thymus, thyroid, and uterus.","In one aspect, an antibody specific for Ras-like GTPase may be used directly as an antagonist, or indirectly as a targeting or delivery mechanism for bringing a pharmaceutical agent to cells or tissue which express Ras-like GTPase.","In another embodiment, a vector expressing the complement of the polynucleotide encoding Ras-like GTPase may be administered to a subject to prevent or treat a cancer including, but not limited to, the types of cancer listed above.","In inflammation, where Ras-like GTPase promotes cell proliferation, it is desirable to decrease its activity.","Therefore, in one embodiment, an antagonist of Ras-like GTPase may be administered to a subject to prevent or treat an inflammation.","Disorders associated with inflammation include, but are not limited to, Addison's disease, adult respiratory distress syndrome, allergies, anemia, asthma, atherosclerosis, bronchitis, cholecystitis, Crohn's disease, ulcerative colitis, atopic dermatitis, dermatomyositis, diabetes mellitus, emphysema, atrophic gastritis, glomerulonephritis, gout, Graves' disease, hypereosinophilia, irritable bowel syndrome, lupus erythematosus, multiple sclerosis, myasthenia gravis, myocardial or pericardial inflammation, osteoarthritis, osteoporosis, pancreatitis, polymyositis, rheumatoid arthritis, scleroderma, Sjogren's syndrome, and autoimmune thyroiditis; complications of cancer, hemodialysis, extracorporeal circulation; viral, bacterial, fungal, parasitic, protozoal, and helminthic infections and trauma.","In one aspect, an antibody specific for Ras-like GTPase may be used directly as an antagonist, or indirectly as a targeting or delivery mechanism for bringing a pharmaceutical agent to cells or tissue which express Ras-like GTPase.","Further, the Ras-like protein polypeptides can be used to screen a compound for the ability to stimulate or inhibit interaction between the Ras-like protein and a molecule that normally interacts with the Ras-like protein, e.g.","a ligand or a component of the signal pathway that the Ras-like protein normally interacts.","Such assays typically include the steps of combining the Ras-like protein with a candidate compound under conditions that allow the Ras-like protein, or fragment, to interact with the target molecule, and to detect the formation of a complex between the protein and the target or to detect the biochemical consequence of the interaction with the Ras-like protein and the target, such as any of the associated effects of signal transduction.","Candidate compounds include, for example, 1) peptides such as soluble peptides, including Ig-tailed fusion peptides and members of random peptide libraries (see, e.g., Lam et al., Nature 354:82-84 (1991); Houghten et al., Nature 354:84-86 (1991)) and combinatorial chemistry-derived molecular libraries made of D- and/or L-configuration amino acids; 2) phosphopeptides (e.g., members of random and partially degenerate, directed phosphopeptide libraries, see, e.g., Songyang et al., Cell 72:767-778 (1993)); 3) antibodies (e.g., polyclonal, monoclonal, humanized, anti-idiotypic, chimeric, and single chain antibodies as well as Fab, F(ab′)2, Fab expression library fragments, and epitope-binding fragments of antibodies); and 4) small organic and inorganic molecules (e.g., molecules obtained from combinatorial and natural product libraries).","(Hodgson, Bio/technology, 1992, Sep. 10(9);973-80).","One candidate compound is a soluble fragment of the Ras-like protein that competes for ligand binding.","Other candidate compounds include mutant Ras-like proteins or appropriate fragments containing mutations that affect Ras-like protein function and thus compete for ligand.","Accordingly, a fragment that competes for ligand, for example with a higher affinity, or a fragment that binds ligand but does not allow release, is within the scope of the invention.","The invention further includes other end point assays to identify compounds that modulate (stimulate or inhibit) Ras-like protein activity.","The assays typically involve an assay of events in the Ras-like protein mediated signal transduction pathway that indicate Ras-like protein activity.","Thus, the phosphorylation of a protein/ligand target, the expression of genes that are up- or down-regulated in response to the Ras-like protein dependent signal cascade can be assayed.","In one embodiment, the regulatory region of such genes can be operably linked to a marker that is easily detectable, such as luciferase.","Alternatively, phosphorylation of the Ras-like protein, or a Ras-like protein target, could also be measured.","Any of the biological or biochemical functions mediated by the Ras-like protein can be used as an endpoint assay.","These include all of the biochemical or biochemical/biological events described herein, in the references cited herein, incorporated by reference for these endpoint assay targets, and other functions known to those of ordinary skill in the art.","Binding and/or activating compounds can also be screened by using chimeric Ras-like proteins in which any of the protein's domains, or parts thereof, can be replaced by heterologous domains or subregions.","Accordingly, a different set of signal transduction components is available as an end-point assay for activation.","This allows for assays to be performed in other than the specific host cell from which the Ras-like protein is derived.","The Ras-like protein polypeptide of the present invention is also useful in competition binding assays in methods designed to discover compounds that interact with the Ras-like protein.","Thus, a compound is exposed to a Ras-like protein polypeptide under conditions that allow the compound to bind or to otherwise interact with the polypeptide.","Soluble Ras-like protein polypeptide is also added to the mixture.","If the test compound interacts with the soluble Ras-like protein polypeptide, it decreases the amount of complex formed or activity from the Ras-like protein target.","This type of assay is particularly useful in cases in which compounds are sought that interact with specific regions of the Ras-like protein.","Thus, the soluble polypeptide that competes with the target Ras-like protein region is designed to contain peptide sequences corresponding to the region of interest.","To perform cell free drug screening assays, it is sometimes desirable to immobilize either the Ras-like protein, or fragment, or its target molecule to facilitate separation of complexes from uncomplexed forms of one or both of the proteins, as well as to accommodate automation of the assay.","Techniques for immobilizing proteins on matrices can be used in the drug screening assays.","In one embodiment, a fusion protein can be provided which adds a domain that allows the protein to be bound to a matrix.","For example, glutathione-S-transferase/15625 fusion proteins can be adsorbed onto glutathione sepharose beads (Sigma Chemical, St. Louis, Mo.)","or glutathione derivatized microtitre plates, which are then combined with the cell lysates (e.g., 35S-labeled) and the candidate compound, and the mixture incubated under conditions conducive to complex formation (e.g., at physiological conditions for salt and pH).","Following incubation, the beads are washed to remove any unbound label, and the matrix immobilized and radiolabel determined directly, or in the supernatant after the complexes are dissociated.","Alternatively, the complexes can be dissociated from the matrix, separated by SDS-PAGE, and the level of Ras-like protein-binding protein found in the bead fraction quantitated from the gel using standard electrophoretic techniques.","For example, either the polypeptide or its target molecule can be immobilized utilizing conjugation of biotin and streptavidin with techniques well known in the art.","Alternatively, antibodies reactive with the protein but which do not interfere with binding of the protein to its target molecule can be derivatized to the wells of the plate, and the protein trapped in the wells by antibody conjugation.","Preparations of a Ras-like protein-binding protein and a candidate compound are incubated in the Ras-like protein-presenting wells and the amount of complex trapped in the well can be quantitated.","Methods for detecting such complexes, in addition to those described above for the GST-immobilized complexes, include immunodetection of complexes using antibodies reactive with the Ras-like protein target molecule, or which are reactive with Ras-like protein and compete with the target molecule, as well as enzyme-linked assays which rely on detecting an enzymatic activity associated with the target molecule.","Agents that modulate one of the Ras-like proteins of the present invention can be identified using one or more of the above assays, alone or in combination.","It is generally preferable to use a cell-based or cell free system first and then confirm activity in an animal/insect model system.","Such model systems are well known in the art and can readily be employed in this context.","Modulators of Ras-like protein activity identified according to these drug screening assays can be used to treat a subject with a disorder mediated by the Ras-like protein associated pathway, by treating cells that express the Ras-like protein.","Experimental data as provided in FIG.","1 indicates expression in bone marrow, stem cells, and leukocytes.","These methods of treatment include the steps of administering the modulators of protein activity in a pharmaceutical composition as described herein, to a subject in need of such treatment.","In yet another aspect of the invention, the Ras-like proteins can be used as “bait proteins” in a two-hybrid assay or three-hybrid assay (see, e.g., U.S. Pat.","No.","5,283,317; Zervos et al., Cell 72:223-232 (1993); Madura et al., J. Biol.","Chem.","268:12046-12054 (1993); Bartel et al., Biotechniques 14:920-924 (1993); Iwabuchi et al., Oncogene 8:1693-1696 (1993); and Brent WO94/10300), to identify other proteins that bind to or interact with the Ras-like protein and are involved in Ras-like protein activity.","Such Ras-like protein-binding proteins are also likely to be involved in the propagation of signals by the Ras-like proteins or Ras-like protein targets as, for example, downstream elements of a Ras-like protein-mediated signaling pathway, e.g., a pain signaling pathway.","Alternatively, such Ras-like protein-binding proteins are likely to be Ras-like protein inhibitors.","The two-hybrid system is based on the modular nature of most transcription factors, which consist of separable DNA-binding and activation domains.","Briefly, the assay utilizes two different DNA constructs.","In one construct, the gene that codes for a Ras-like protein is fused to a gene encoding the DNA binding domain of a known transcription factor (e.g., GAL-4).","In the other construct, a DNA sequence, from a library of DNA sequences, that encodes an unidentified protein (“prey” or “sample”) is fused to a gene that codes for the activation domain of the known transcription factor.","If the “bait” and the “prey” proteins are able to interact, in vivo, forming a Ras-like protein-dependent complex, the DNA-binding and activation domains of the transcription factor are brought into close proximity.","This proximity allows transcription of a reporter gene (e.g., LacZ) which is operably linked to a transcriptional regulatory site responsive to the transcription factor.","Expression of the reporter gene can be detected and cell colonies containing the functional transcription factor can be isolated and used to obtain the cloned gene which encodes the protein which interacts with the Ras-like protein.","This invention further pertains to novel agents identified by the above-described screening assays.","Accordingly, it is within the scope of this invention to further use an agent identified as described herein in an appropriate animal model.","For example, an agent identified as described herein (e.g., a Ras-like protein modulating agent, an antisense Ras-like protein nucleic acid molecule, a Ras-like protein-specific antibody, or a Ras-like protein-binding partner) can be used in an animal or insect model to determine the efficacy, toxicity, or side effects of treatment with such an agent.","Alternatively, an agent identified as described herein can be used in an animal or insect model to determine the mechanism of action of such an agent.","Furthermore, this invention pertains to uses of novel agents identified by the above-described screening assays for treatments as described herein.","The Ras-like proteins of the present invention are also useful to provide a target for diagnosing a disease or predisposition to a disease mediated by the peptide, Accordingly, the invention provides methods for detecting the presence, or levels of, the protein (or encoding mRNA) in a cell, tissue, or organism.","Experimental data as provided in FIG.","1 indicates expression in bone marrow, stem cells, and leukocytes.","The method involves contacting a biological sample with a compound capable of interacting with the receptor protein such that the interaction can be detected.","Such an assay can be provided in a single detection format or a multi-detection format such as an antibody chip array.","One agent for detecting a protein in a sample is an antibody capable of selectively binding to protein.","A biological sample includes tissues, cells and biological fluids isolated from a subject, as well as tissues, cells, and fluids present within a subject.","The peptides also are useful to provide a target for diagnosing a disease or predisposition to a disease mediated by the peptide, Accordingly, the invention provides methods for detecting the presence, or levels of, the protein in a cell, tissue, or organism.","The method involves contacting a biological sample with a compound capable of interacting with the receptor protein such that the interaction can be detected.","The peptides of the present invention also provide targets for diagnosing active disease, or predisposition to a disease, in a patient having a variant peptide.","Thus, the peptide can be isolated from a biological sample and assayed for the presence of a genetic mutation that results in translation of an aberrant peptide.","This includes amino acid substitution, deletion, insertion, rearrangement, (as the result of aberrant splicing events), and inappropriate post-translational modification.","Analytic methods include altered electrophoretic mobility, altered tryptic peptide digest, altered receptor activity in cell-based or cell-free assay, alteration in ligand or antibody-binding pattern, altered isoelectric point, direct amino acid sequencing, and any other of the known assay techniques useful for detecting mutations in a protein.","Such an assay can be provided in a single detection format or a multi-detection format such as an antibody chip array.","In vitro techniques for detection of peptide include enzyme linked immunosorbent assays (ELISAs), Western blots, immunoprecipitations, and immunofluorescence using a detection reagents, such as an antibody or protein binding agent.","Alternatively, the peptide can be detected in vivo in a subject by introducing into the subject a labeled anti-peptide antibody.","For example, the antibody can be labeled with a radioactive marker whose presence and location in a subject can be detected by standard imaging techniques.","Particularly useful are methods that detect the allelic variant of a peptide expressed in a subject and methods which detect fragments of a peptide in a sample.","The peptides are also useful in pharmacogenomic analysis.","Pharmacogenomics deal with clinically significant hereditary variations in the response to drugs due to altered drug disposition and abnormal action in affected persons.","See, e.g., Eichelbaum, M. (Clin.","Exp.","Pharmacol.","Physiol.","23(10-11):983-985 (1996)), and Linder, M. W. (Clin.","Chem.","43(2):254-266 (1997)).","The clinical outcomes of these variations result in severe toxicity of therapeutic drugs in certain individuals or therapeutic failure of drugs in certain individuals as a result of individual variation in metabolism.","Thus, the genotype of the individual can determine the way a therapeutic compound acts on the body or the way the body metabolizes the compound.","Further, the activity of drug metabolizing enzymes effects both the intensity and duration of drug action.","Thus, the pharmacogenomics of the individual permit the selection of effective compounds and effective dosages of such compounds for prophylactic or therapeutic treatment based on the individual's genotype.","The discovery of genetic polymorphisms in some drug metabolizing enzymes has explained why some patients do not obtain the expected drug effects, show an exaggerated drug effect, or experience serious toxicity from standard drug dosages.","Polymorphisms can be expressed in the phenotype of the extensive metabolizer and the phenotype of the poor metabolizer.","Accordingly, genetic polymorphism may lead to allelic protein variants of the receptor protein in which one or more of the receptor functions in one population is different from those in another population.","The peptides thus allow a target to ascertain a genetic predisposition that can affect treatment modality.","Thus, in a ligand-based treatment, polymorphism may give rise to amino terminal extracellular domains and/or other ligand-binding regions that are more or less active in ligand binding, and receptor activation.","Accordingly, ligand dosage would necessarily be modified to maximize the therapeutic effect within a given population containing a polymorphism.","As an alternative to genotyping, specific polymorphic peptides could be identified.","The peptides are also useful for treating a disorder characterized by an absence of, inappropriate, or unwanted expression of the protein.","Experimental data as provided in FIG.","1 indicates expression in bone marrow, stem cells, and leukocytes.","Accordingly, methods for treatment include the use of the Ras-like protein or fragments.","Antibodies The invention also provides antibodies that selectively bind to one of the peptides of the present invention, a protein comprising such a peptide, as well as variants and fragments thereof.","As used herein, an antibody selectively binds a target peptide when it binds the target peptide and does not significantly bind to unrelated proteins.","An antibody is still considered to selectively bind a peptide even if it also binds to other proteins that are not substantially homologous with the target peptide so long as such proteins share homology with a fragment or domain of the peptide target of the antibody.","In this case, it would be understood that antibody binding to the peptide is still selective despite some degree of cross-reactivity.","As used herein, an antibody is defined in terrns consistent with that recognized within the art: they are multi-subunit proteins produced by a mammalian organism in response to an antigen challenge.","The antibodies of the present invention include polyclonal antibodies and monoclonal antibodies, as well as fragments of such antibodies, including, but not limited to, Fab or F(ab′)2, and Fv fragments.","Many methods are known for generating and/or identifying antibodies to a given target peptide.","Several such methods are described by Harlow, Antibodies, Cold Spring Harbor Press, (1989).","In general, to generate antibodies, an isolated peptide is used as an immunogen and is administered to a mammalian organism, such as a rat, rabbit or mouse.","The full-length protein, an antigenic peptide fragment or a fusion protein can be used.","Particularly important fragments are those covering functional domains, such as the domains identified in FIG.","2, and domain of sequence homology or divergence amongst the family, such as those that can readily be identified using protein alignment methods and as presented in the Figures.","Antibodies are preferably prepared from regions or discrete fragments of the Ras-like proteins.","Antibodies can be prepared from any region of the peptide as described herein.","However, preferred regions will include those involved in function/activity and/or receptor/binding partner interaction.","FIG.","2 can be used to identify particularly important regions while sequence alignment can be used to identify conserved and unique sequence fragments.","An antigenic fragment will typically comprise at least 8 contiguous amino acid residues.","The antigenic peptide can comprise, however, at least 10, 12, 14, 16 or more amino acid residues.","Such fragments can be selected on a physical property, such as fragments correspond to regions that are located on the surface of the protein, e.g., hydrophilic regions or can be selected based on sequence uniqueness (see FIG.","2).","Detection of an antibody of the present invention can be facilitated by coupling (i.e., physically linking) the antibody to a detectable substance.","Examples of detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, and radioactive materials.","Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, β-galactosidase, or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidin/biotin and avidin/biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; an example of a luminescent material includes luminol; examples of bioluminescent materials include luciferase, luciferin, and aequorin, and examples of suitable radioactive material include 125I, 131I, 35S, or 3H.","Antibody Uses The antibodies can be used to isolate one of the proteins of the present invention by standard techniques, such as affinity chromatography or immunoprecipitation.","The antibodies can facilitate the purification of the natural protein from cells and recombinantly produced protein expressed in host cells.","In addition, such antibodies are useful to detect the presence of one of the proteins of the present invention in cells or tissues to determine the pattern of expression of the protein among various tissues in an organism and over the course of normal development.","Experimental data as provided in FIG.","1 indicates that Ras-like proteins of the present invention are expressed in bone marrow and stem cells (as indicated by virtual northern blot analysis), and leukocytes (as indicated by PCR-based tissue screening panels).","Further, such antibodies can be used to detect protein in situ, in vitro, or in a cell lysate or supernatant in order to evaluate the abundance and pattern of expression.","Also, such antibodies can be used to assess abnormal tissue distribution or abnormal expression during development.","Antibody detection of circulating fragments of the full-length protein can be used to identify turnover.","Further, the antibodies can be used to assess expression in disease states such as in active stages of the disease or in an individual with a predisposition toward disease related to the protein's function.","When a disorder is caused by an inappropriate tissue distribution, developmental expression, level of expression of the protein, or expressed/processed form, the antibody can be prepared against the normal protein.","Experimental data as provided in FIG.","1 indicates expression in bone marrow, stem cells, and leukocytes.","If a disorder is characterized by a specific mutation in the protein, antibodies specific for this mutant protein can be used to assay for the presence of the specific mutant protein.","The antibodies can also be used to assess normal and aberrant subcellular localization of cells in the various tissues in an organism.","Experimental data as provided in FIG.","1 indicates expression in bone marrow, stem cells, and leukocytes.","The diagnostic uses can be applied, not only in genetic testing, but also in monitoring a treatment modality.","Accordingly, where treatment is ultimately aimed at correcting expression level or the presence of aberrant sequence and aberrant tissue distribution or developmental expression, antibodies directed against the or relevant fragments can be used to monitor therapeutic efficacy.","Additionally, antibodies are useful in pharmacogenomic analysis.","Thus, antibodies prepared against polymorphic proteins can be used to identify individuals that require modified treatment modalities.","The antibodies are also useful as diagnostic tools as an immunological marker for aberrant protein analyzed by electrophoretic mobility, isoelectric point, tryptic peptide digest, and other physical assays known to those in the art.","The antibodies are also useful for tissue typing.","Experimental data as provided in FIG.","1 indicates expression in bone marrow, stem cells, and leukocytes.","Thus, where a specific protein has been correlated with expression in a specific tissue, antibodies that are specific for this protein can be used to identify a tissue type.","The antibodies are also useful for inhibiting protein function, for example, blocking the binding of the Ras-like protein to a binding partner such as a substrate.","These uses can also be applied in a therapeutic context in which treatment involves inhibiting the protein's function.","An antibody can be used, for example, to block binding, thus modulating (agonizing or antagonizing) the peptides activity.","Antibodies can be prepared against specific fragments containing sites required for function or against intact protein that is associated with a cell or cell membrane.","See FIG.","2 for structural information relating to the proteins of the present invention.","The invention also encompasses kits for using antibodies to detect the presence of a protein in a biological sample.","The kit can comprise antibodies such as a labeled or labelable antibody and a compound or agent for detecting protein in a biological sample; means for determining the amount of protein in the sample; means for comparing the amount of protein in the sample with a standard; and instructions for use.","Nucleic Acid Molecules The present invention further provides isolated nucleic acid molecules that encode a Ras-like protein polypeptide of the present invention.","Such nucleic acid molecules will consist of, consist essentially of, or comprise a nucleotide sequence that encodes one of the Ras-like protein polypeptides of the present invention, an allelic variant thereof, or an ortholog or paralog thereof.","As used herein, an “isolated” nucleic acid molecule is one that is separated from other nucleic acid present in the natural source of the nucleic acid.","Preferably, an “isolated” nucleic acid is free of sequences which naturally flank the nucleic acid (i.e., sequences located at the 5′ and 3′ ends of the nucleic acid) in the genomic DNA of the organism from which the nucleic acid is derived.","However, there can be some flanking nucleotide sequences, for example up to about 5 KB, particularly contiguous peptide encoding sequences and peptide encoding sequences within the same gene but separated by introns in the genomic sequence.","The important point is that the nucleic acid is isolated from remote and unimportant flanking sequences such that it can be subjected to the specific manipulations described herein such as recombinant expression, preparation of probes and primers, and other uses specific to the nucleic acid sequences.","Moreover, an “isolated” nucleic acid molecule, such as a cDNA molecule, can be substantially free of other cellular material, or culture medium when produced by recombinant techniques, or chemical precursors or other chemicals when chemically synthesized.","However, the nucleic acid molecule can be fused to other coding or regulatory sequences and still be considered isolated.","For example, recombinant DNA molecules contained in a vector are considered isolated.","Further examples of isolated DNA molecules include recombinant DNA molecules maintained in heterologous host cells or purified (partially or substantially) DNA molecules in solution.","Isolated RNA molecules include in vivo or in vitro RNA transcripts of the isolated DNA molecules of the present invention.","Isolated nucleic acid molecules according to the present invention further include such molecules produced synthetically.","Accordingly, the present invention provides nucleic acid molecules that consist of the nucleotide sequence shown in FIG.","1 or 3 (SEQ ID NO:1, transcript sequence and SEQ ID NO:3, genomic sequence), or any nucleic acid molecule that encodes the protein provided in FIG.","2, SEQ ID NO:2.A nucleic acid molecule consists of a nucleotide sequence when the nucleotide sequence is the complete nucleotide sequence of the nucleic acid molecule.","The present invention further provides nucleic acid molecules that consist essentially of the nucleotide sequence shown in FIG.","1 or 3 (SEQ ID NO:1, transcript sequence and SEQ ID NO:3, genomic sequence), or any nucleic acid molecule that encodes the protein provided in FIG.","2, SEQ ID NO:2.A nucleic acid molecule consists essentially of a nucleotide sequence when such a nucleotide sequence is present with only a few additional nucleic acid residues in the final nucleic acid molecule.","The present invention further provides nucleic acid molecules that comprise the nucleotide sequences shown in FIG.","1 or 3 (SEQ ID NO:1, transcript sequence and SEQ ID NO:3, genomic sequence), or any nucleic acid molecule that encodes the protein provided in FIG.","2, SEQ ID NO:2.A nucleic acid molecule comprises a nucleotide sequence when the nucleotide sequence is at least part of the final nucleotide sequence of the nucleic acid molecule.","In such a fashion, the nucleic acid molecule can be only the nucleotide sequence or have additional nucleic acid residues, such as nucleic acid residues that are naturally associated with it or heterologous nucleotide sequences.","Such a nucleic acid molecule can have a few additional nucleotides or can comprises several hundred or more additional nucleotides.","A brief description of how various types of these nucleic acid molecules can be readily made/isolated is provided below.","In FIGS.","1 and 3, both coding and non-coding sequences are provided.","Because of the source of the present invention, humans genomic sequence (FIG.","3) and cDNA/transcript sequences (FIG.","1), the nucleic acid molecules in the Figures will contain genomic intronic sequences, 5′ and 3′ non-coding sequences, gene regulatory regions and non-coding intergenic sequences.","In general such sequence features are either noted in FIGS.","1 and 3 or can readily be identified using computational tools known in the art.","As discussed below, some of the non-coding regions, particularly gene regulatory elements such as promoters, are useful for a variety of purposes, e.g.","control of heterologous gene expression, target for identifying gene activity modulating compounds, and are particularly claimed as fragments of the genomic sequence provided herein.","Full-length genes may be cloned from known sequence using any one of a number of methods known in the art.","For example, a method which employs XL-PCR (Perkin-Elmer, Foster City, Calif.) to amplify long pieces of DNA may be used.","Other methods for obtaining full-length sequences are well known in the art.","The isolated nucleic acid molecules can encode the mature protein plus additional amino or carboxyl-terminal amino acids, or amino acids interior to the mature peptide (when the mature form has more than one peptide chain, for instance).","Such sequences may play a role in processing of a protein from precursor to a mature form, facilitate protein trafficking, prolong or shorten protein half-life, or facilitate manipulation of a protein for assay or production, among other things.","As generally is the case in situ, the additional amino acids may be processed away from the mature protein by cellular enzymes.","As mentioned above, the isolated nucleic acid molecules include, but are not limited to, the sequence encoding the Ras-like protein polypeptide alone, the sequence encoding the mature peptide and additional coding sequences, such as a leader or secretory sequence (e.g., a pre-pro or pro-protein sequence), the sequence encoding the mature peptide, with or without the additional coding sequences, plus additional non-coding sequences, for example introns and non-coding 5′ and 3′ sequences such as transcribed but non-translated sequences that play a role in transcription, mRNA processing (including splicing and polyadenylation signals), ribosome binding, and stability of mRNA.","In addition, the nucleic acid molecule may be fused to a marker sequence encoding, for example, a peptide that facilitates purification.","Isolated nucleic acid molecules can be in the form of RNA, such as mRNA, or in the form of DNA, including cDNA and genomic DNA obtained by cloning or produced by chemical synthetic techniques or by a combination thereof.","The nucleic acid, especially DNA, can be double-stranded or single-stranded.","Single-stranded nucleic acid can be the coding strand (sense strand) or the non-coding strand (anti-sense strand).","The invention further provides nucleic acid molecules that encode fragments of the peptides of the present invention and that encode obvious variants of the Ras-like proteins of the present invention that are described above.","Such nucleic acid molecules may be naturally occurring, such as allelic variants (same locus), paralogs (different locus), and orthologs (different organism), or may be constructed by recombinant DNA methods or by chemical synthesis.","Such non-naturally occurring variants may be made by mutagenesis techniques, including those applied to nucleic acid molecules, cells, or whole organisms.","Accordingly, as discussed above, the variants can contain nucleotide substitutions, deletions inversions, and/or insertions.","Variation can occur in either or both the coding and non-coding regions.","The variations can produce both conservative and non-conservative amino acid substitutions.","The present invention further provides non-coding fragments of the nucleic acid molecules provided in the FIGS.","1 and 3.Preferred non-coding fragments include, but are not limited to, promoter sequences, enhancer sequences, gene modulating sequences, and gene termination sequences.","Such fragments are useful in controlling heterologous gene expression and in developing screens to identify gene-modulating agents.","A fragment comprises a contiguous nucleotide sequence greater than 12 or more nucleotides.","Further, a fragment could be at least 30, 40, 50, 100 250, or 500 nucleotides in length.","The length of the fragment will be based on its intended use.","For example, the fragment can encode epitope-bearing regions of the peptide, or can be useful as DNA probes and primers.","Such fragments can be isolated using the known nucleotide sequence to synthesize an oligonucleotide probe.","A labeled probe can then be used to screen a cDNA library, genomic DNA library, or mRNA to isolate nucleic acid corresponding to the coding region.","Further, primers can be used in PCR reactions to clone specific regions of gene.","A probe/primer typically comprises substantially a purified oligonucleotide or oligonucleotide pair.","The oligonucleotide typically comprises a region of nucleotide sequence that hybridizes under stringent conditions to at least about 12, 20, 25, 40, 50, or more consecutive nucleotides.","Orthologs, homologs, and allelic variants can be identified using methods well known in the art.","As described in the Peptide Section, these variants comprise a nucleotide sequence encoding a peptide that is typically 60-70%, 70-80%, 80-90%, and more typically at least about 90-95% or more homologous to the nucleotide sequence shown in the Figure sheets or a fragment of this sequence.","Such nucleic acid molecules can readily be identified as being able to hybridize under moderate to stringent conditions, to the nucleotide sequence shown in the Figure sheets or a fragment of the sequence.","The gene encoding the novel Ras-like protein of the present invention is located on a genome component that has been mapped to human chromosome 6 (as indicated in FIG.","3), which is supported by multiple lines of evidence, such as STS and BAC map data.","FIG.","3 provides information on SNPs that have been found in the gene encoding the Ras-like protein of the present invention.","SNPs were identified at 37 different nucleotide positions.","Some of these SNPs that are located outside the ORF and in introns may affect gene transcription.","As used herein, the term “hybridizes under stringent conditions” is intended to describe conditions for hybridization and washing under which nucleotide sequences encoding a peptide at least 60-70% homologous to each other typically remain hybridized to each other.","The conditions can be such that sequences at least about 60%, at least about 70%, or at least about 80% or more homologous to each other typically remain hybridized to each other.","Such stringent conditions are known to those skilled in the art and can be found in Current Protocols in Molecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6.One example of stringent hybridization conditions are hybridization in 6× sodium chloridelsodium citrate (SSC) at about 45 C, followed by one or more washes in 0.2×SSC, 0.1% SDS at 50-65° C. Examples of moderate to low stringency hybridization conditions are well known in the art.","Nucleic Acid Molecule Uses The nucleic acid molecules of the present invention are useful for probes, primers, chemical intermediates, and in biological assays.","The nucleic acid molecules are useful as a hybridization probe for messenger RNA, transcript/cDNA and genomic DNA to isolate full-length cDNA and genomic clones encoding the peptide described in FIG.","2 and to isolate cDNA and genomic clones that correspond to variants (alleles, orthologs, etc.)","producing the same or related peptides shown in FIG.","2.As illustrated in FIG.","3, SNPs were identified at 37 different nucleotide positions.","The probe can correspond to any sequence along the entire length of the nucleic acid molecules provided in the Figures.","Accordingly, it could be derived from 5′ noncoding regions, the coding region, and 3′ noncoding regions.","However, as discussed, fragments are not to be construed as those, which may encompass fragments disclosed prior to the present invention.","The nucleic acid molecules are also useful as primers for PCR to amplify any given region of a nucleic acid molecule and are useful to synthesize antisense molecules of desired length and sequence.","The nucleic acid molecules are also useful for constructing recombinant vectors.","Such vectors include expression vectors that express a portion of, or all of, the peptide sequences.","Vectors also include insertion vectors, used to integrate into another nucleic acid molecule sequence, such as into the cellular genome, to alter in situ expression of a gene and/or gene product.","For example, an endogenous coding sequence can be replaced via homologous recombination with all or part of the coding region containing one or more specifically introduced mutations.","The nucleic acid molecules are also useful for expressing antigenic portions of the proteins.","The nucleic acid molecules are also useful as probes for determining the chromosomal positions of the nucleic acid molecules by means of in situ hybridization methods.","The gene encoding the novel Ras-like protein of the present invention is located on a genome component that has been mapped to human chromosome 6 (as indicated in FIG.","3), which is supported by multiple lines of evidence, such as STS and BAC map data.","The nucleic acid molecules are also useful in making vectors containing the gene regulatory regions of the nucleic acid molecules of the present invention.","The nucleic acid molecules are also useful for designing ribozymes corresponding to all, or a part, of the mRNA produced from the nucleic acid molecules described herein.","The nucleic acid molecules are also useful for constructing host cells expressing a part, or all, of the nucleic acid molecules and peptides.","Moreover, the nucleic acid molecules are useful for constructing transgenic animals wherein a homolog of the nucleic acid molecule has been “knocked-out” of the animal's genome.","The nucleic acid molecules are also useful for constructing transgenic animals expressing all, or a part, of the nucleic acid molecules and peptides.","The nucleic acid molecules are also useful for making vectors that express part, or all, of the peptides.","The nucleic acid molecules are also useful as hybridization probes for determining the presence, level, form, and distribution of nucleic acid expression.","Experimental data as provided in FIG.","1 indicates that Ras-like proteins of the present invention are expressed in bone marrow and stem cells (as indicated by virtual northern blot analysis), and leukocytes (as indicated by PCR-based tissue screening panels).","Accordingly, the probes can be used to detect the presence of, or to determine levels of, a specific nucleic acid molecule in cells, tissues, and in organisms.","The nucleic acid whose level is determined can be DNA or RNA.","Accordingly, probes corresponding to the peptides described herein can be used to assess expression and/or gene copy number in a given cell, tissue, or organism.","These uses are relevant for diagnosis of disorders involving an increase or decrease in Ras-like protein expression relative to normal results.","In vitro techniques for detection of mRNA include Northern hybridizations and in situ hybridizations.","In vitro techniques for detecting DNA include Southern hybridizations and in situ hybridization.","Probes can be used as a part of a diagnostic test kit for identifying cells or tissues that express a Ras-like protein, such as by measuring a level of a receptor-encoding nucleic acid in a sample of cells from a subject e.g., mRNA or genomic DNA, or determining if a receptor gene has been mutated.","Experimental data as provided in FIG.","1 indicates that Ras-like proteins of the present invention are expressed in bone marrow and stem cells (as indicated by virtual northern blot analysis), and leukocytes (as indicated by PCR-based tissue screening panels).","Nucleic acid expression assays are useful for drug screening to identify compounds that modulate Ras-like protein nucleic acid expression.","The invention thus provides a method for identifying a compound that can be used to treat a disorder associated with nucleic acid expression of the Ras-like protein gene, particularly biological and pathological processes that are mediated by the Ras-like protein in cells and tissues that express it.","Experimental data as provided in FIG.","1 indicates expression in bone marrow, stem cells, and leukocytes.","The method typically includes assaying the ability of the compound to modulate the expression of the Ras-like protein nucleic acid and thus identifying a compound that can be used to treat a disorder characterized by undesired Ras-like protein nucleic acid expression.","The assays can be performed in cell-based and cell-free systems.","Cell-based assays include cells naturally expressing the Ras-like protein nucleic acid or recombinant cells genetically engineered to express specific nucleic acid sequences.","The assay for Ras-like protein nucleic acid expression can involve direct assay of nucleic acid levels, such as mRNA levels, or on collateral compounds involved in the signal pathway.","Further, the expression of genes that are up- or down-regulated in response to the Ras-like protein signal pathway can also be assayed.","In this embodiment the regulatory regions of these genes can be operably linked to a reporter gene such as luciferase.","Thus, modulators of Ras-like protein gene expression can be identified in a method wherein a cell is contacted with a candidate compound and the expression of mRNA determined.","The level of expression of Ras-like protein mRNA in the presence of the candidate compound is compared to the level of expression of Ras-like protein mRNA in the absence of the candidate compound.","The candidate compound can then be identified as a modulator of nucleic acid expression based on this comparison and be used, for example to treat a disorder characterized by aberrant nucleic acid expression.","When expression of mRNA is statistically significantly greater in the presence of the candidate compound than in its absence, the candidate compound is identified as a stimulator of nucleic acid expression.","When nucleic acid expression is statistically significantly less in the presence of the candidate compound than in its absence, the candidate compound is identified as an inhibitor of nucleic acid expression.","The invention further provides methods of treatment, with the nucleic acid as a target, using a compound identified through drug screening as a gene modulator to modulate Ras-like protein nucleic acid expression in cells and tissues that express the Ras-like protein.","Experimental data as provided in FIG.","1 indicates that Ras-like proteins of the present invention are expressed in bone marrow and stem cells (as indicated by virtual northern blot analysis), and leukocytes (as indicated by PCR-based tissue screening panels).","Modulation includes both up-regulation (i.e.","activation or agonization) or down-regulation (suppression or antagonization) of nucleic acid expression.","Alternatively, a modulator for Ras-like protein nucleic acid expression can be a small molecule or drug identified using the screening assays described herein as long as the drug or small molecule inhibits the Ras-like protein nucleic acid expression in the cells and tissues that express the protein.","Experimental data as provided in FIG.","1 indicates expression in bone marrow, stem cells, and leukocytes.","The nucleic acid molecules are also useful for monitoring the effectiveness of modulating compounds on the expression or activity of the Ras-like protein gene in clinical trials or in a treatment regimen.","Thus, the gene expression pattern can serve as a barometer for the continuing effectiveness of treatment with the compound, particularly with compounds to which a patient can develop resistance.","The gene expression pattern can also serve as a marker indicative of a physiological response of the affected cells to the compound.","Accordingly, such monitoring would allow either increased administration of the compound or the administration of alternative compounds to which the patient has not become resistant.","Similarly, if the level of nucleic acid expression falls below a desirable level, administration of the compound could be commensurately decreased.","The nucleic acid molecules are also useful in diagnostic assays for qualitative changes in Ras-like protein nucleic acid, and particularly in qualitative changes that lead to pathology.","The nucleic acid molecules can be used to detect mutations in Ras-like protein genes and gene expression products such as mRNA.","The nucleic acid molecules can be used as hybridization probes to detect naturally occurring genetic mutations in the Ras-like protein gene and thereby to determine whether a subject with the mutation is at risk for a disorder caused by the mutation.","Mutations include deletion, addition, or substitution of one or more nucleotides in the gene, chromosomal rearrangement, such as inversion or transposition, modification of genomic DNA, such as aberrant methylation patterns, or changes in gene copy number, such as amplification.","Detection of a mutated form of the Ras-like protein gene associated with a dysfunction provides a diagnostic tool for an active disease or susceptibility to disease when the disease results from overexpression, underexpression, or altered expression of a Ras-like protein.","Individuals carrying mutations in the Ras-like protein gene can be detected at the nucleic acid level by a variety of techniques.","FIG.","3 provides information on SNPs that have been found in the gene encoding the Ras-like protein of the present invention.","SNPs were identified at 37 different nucleotide positions.","Some of these SNPs that are located outside the ORF and in introns may affect gene transcription.","The gene encoding the novel Ras-like protein of the present invention is located on a genome component that has been mapped to human chromosome 6 (as indicated in FIG.","3), which is supported by multiple lines of evidence, such as STS and BAC map data.","Genomic DNA can be analyzed directly or can be amplified by using PCR prior to analysis.","RNA or cDNA can be used in the same way.","In some uses, detection of the mutation involves the use of a probe/primer in a polymerase chain reaction (PCR) (see, e.g.","U.S. Pat.","Nos.","4,683,195 and 4,683,202), such as anchor PCR or RACE PCR, or, alternatively, in a ligation chain reaction (LCR) (see, e.g., Landegran et al., Science 241:1077-1080 (1988); and Nakazawa et al., PNAS 91:360-364 (1994)), the latter of which can be particularly useful for detecting point mutations in the gene (see Abravaya et al., Nucleic Acids Res.","23:675-682 (1995)).","This method can include the steps of collecting a sample of cells from a patient, isolating nucleic acid (e.g., genomic, mRNA or both) from the cells of the sample, contacting the nucleic acid sample with one or more primers which specifically hybridize to a gene under conditions such that hybridization and amplification of the gene (if present) occurs, and detecting the presence or absence of an amplification product, or detecting the size of the amplification product and comparing the length to a control sample.","Deletions and insertions can be detected by a change in size of the amplified product compared to the normal genotype.","Point mutations can be identified by hybridizing amplified DNA to normal RNA or antisense DNA sequences.","Alternatively, mutations in a Ras-like protein gene can be directly identified, for example, by alterations in restriction enzyme digestion patterns determined by gel electrophoresis.","Further, sequence-specific ribozymes (U.S. Pat.","No.","5,498,531) can be used to score for the presence of specific mutations by development or loss of a ribozyme cleavage site.","Perfectly matched sequences can be distinguished from mismatched sequences by nuclease cleavage digestion assays or by differences in melting temperature.","Sequence changes at specific locations can also be assessed by nuclease protection assays such as RNase and S1 protection or the chemical cleavage method.","Furthermore, sequence differences between a mutant Ras-like protein gene and a wild-type gene can be determined by direct DNA sequencing.","A variety of automated sequencing procedures can be utilized when performing the diagnostic assays (Naeve, C. W., Biotechniques 19:448 (1995)), including sequencing by mass spectrometry (see, e.g., PCT International Publication No.","WO 94/16101; Cohen et al., Adv.","Chromatogr.","36:127-162 (1996); and Griffin et al., Appl.","Biochem.","Biotechnol.","38:147-159 (1993)).","Other methods for detecting mutations in the gene include methods in which protection from cleavage agents is used to detect mismatched bases in RNA/RNA or RNA/DNA duplexes (Myers et al., Science 230:1242 (1985)); Cotton et al., PNAS 85:4397 (1988); Saleeba et al., Meth.","Enzymol.","217:286-295 (1992)), electrophoretic mobility of mutant and wild type nucleic acid is compared (Orita et al., PNAS86:2766 (1989); Cotton et al., Mutat.","Res.","285:125-144 (1993); and Hayashi et al., Genet.","Anal.","Tech Appl.","9:73-79 (1992)), and movement of mutant or wild-type fragments in polyacrylamide gels containing a gradient of denaturant is assayed using denaturing gradient gel electrophoresis (Myers et al., Nature 313:495 (1985)).","Examples of other techniques for detecting point mutations include, selective oligonucleotide hybridization, selective amplification, and selective primer extension.","The nucleic acid molecules are also useful for testing an individual for a genotype that while not necessarily causing the disease, nevertheless affects the treatment modality.","Thus, the nucleic acid molecules can be used to study the relationship between an individual's genotype and the individual's response to a compound used for treatment (pharmacogenomic relationship).","Accordingly, the nucleic acid molecules described herein can be used to assess the mutation content of the Ras-like protein gene in an individual in order to select an appropriate compound or dosage regimen for treatment.","FIG.","3 provides information on SNPs that have been found in the gene encoding the Ras-like protein of the present invention.","SNPs were identified at 37 different nucleotide positions.","Some of these SNPs that are located outside the ORF and in introns may affect gene transcription.","Thus nucleic acid molecules displaying genetic variations that affect treatment provide a diagnostic target that can be used to tailor treatment in an individual.","Accordingly, the production of recombinant cells and animals containing these polymorphisms allow effective clinical design of treatment compounds and dosage regimens.","The nucleic acid molecules are thus useful as antisense constructs to control Ras-like protein gene expression in cells, tissues, and organisms.","A DNA antisense nucleic acid molecule is designed to be complementary to a region of the gene involved in transcription, preventing transcription and hence production of Ras-like protein.","An antisense RNA or DNA nucleic acid molecule would hybridize to the mRNA and thus block translation of mRNA into Ras-like protein.","Alternatively, a class of antisense molecules can be used to inactivate mRNA in order to decrease expression of Ras-like protein nucleic acid.","Accordingly, these molecules can treat a disorder characterized by abnormal or undesired Ras-like protein nucleic acid expression.","This technique involves cleavage by means of ribozymes containing nucleotide sequences complementary to one or more regions in the mRNA that attenuate the ability of the mRNA to be translated.","Possible regions include coding regions and particularly coding regions corresponding to the catalytic and other functional activities of the Ras-like protein, such as ligand binding.","The nucleic acid molecules also provide vectors for gene therapy in patients containing cells that are aberrant in Ras-like protein gene expression.","Thus, recombinant cells, which include the patient's cells that have been engineered ex vivo and returned to the patient, are introduced into an individual where the cells produce the desired Ras-like protein to treat the individual.","The invention also encompasses kits for detecting the presence of a Ras-like protein nucleic acid in a biological sample.","Experimental data as provided in FIG.","1 indicates that Ras-like proteins of the present invention are expressed in bone marrow and stem cells (as indicated by virtual northern blot analysis), and leukocytes (as indicated by PCR-based tissue screening panels).","For example, the kit can comprise reagents such as a labeled or labelable nucleic acid or agent capable of detecting Ras-like protein nucleic acid in a biological sample; means for determining the amount of Ras-like protein nucleic acid in the sample; and means for comparing the amount of Ras-like protein nucleic acid in the sample with a standard.","The compound or agent can be packaged in a suitable container.","The kit can further comprise instructions for using the kit to detect Ras-like protein mRNA or DNA.","Nucleic Acid Arrays The present invention further provides arrays or microarrays of nucleic acid molecules that are based on the sequence information provided in FIGS.","1 and 3 (SEQ ID NOS:1 and 3).","As used herein “Arrays” or “Microarrays” refers to an array of distinct polynucleotides or oligonucleotides synthesized on a substrate, such as paper, nylon or other type of membrane, filter, chip, glass slide, or any other suitable solid support.","In one embodiment, the microarray is prepared and used according to the methods described in U.S. Pat.","No.","5,837,832, Chee et al., PCT application WO95/11995 (Chee et al.","), Lockhart, D. J. et al.","(1996; Nat.","Biotech.","14: 1675-1680) and Schena, M. et al.","(1996; Proc.","Natl.","Acad.","Sci.","93: 10614-10619), all of which are incorporated herein in their entirety by reference.","In other embodiments, such arrays are produced by the methods described by Brown et.","al., U.S. Pat.","No.","5,807,522.The microarray is preferably composed of a large number of unique, single-stranded nucleic acid sequences, usually either synthetic antisense oligonucleotides or fragments of cDNAs, fixed to a solid support.","The oligonucleotides are preferably about 6-60 nucleotides in length, more preferably 15-30 nucleotides in length, and most preferably about 20-25 nucleotides in length.","For a certain type of microarray, it may be preferable to use oligonucleotides that are only 7-20 nucleotides in length.","The microarray may contain oligonucleotides that cover the known 5′, or 3′, sequence, sequential oligonucleotides that cover the full-length sequence; or unique oligonucleotides selected from particular areas along the length of the sequence.","Polynucleotides used in the microarray may be oligonucleotides that are specific to a gene or genes of interest.","In order to produce oligonucleotides to a known sequence for a microarray, the gene(s) of interest (or an ORF identified from the contigs of the present invention) is typically examined using a computer algorithm that starts at the 5′ or at the 3′ end of the nucleotide sequence.","Typical algorithms will then identify oligomers of defined length that are unique to the gene, have a GC content within a range suitable for hybridization, and lack predicted secondary structure that may interfere with hybridization.","In certain situations it may be appropriate to use pairs of oligonucleotides on a microarray.","The “pairs” will be identical, except for one nucleotide that preferably is located in the center of the sequence.","The second oligonucleotide in the pair (mismatched by one) serves as a control.","The number of oligonucleotide pairs may range from two to one million.","The oligomers are synthesized at designated areas on a substrate using a light-directed chemical process.","The substrate may be paper, nylon or other type of membrane, filter, chip, glass slide or any other suitable solid support.","In another aspect, an oligonucleotide may be synthesized on the surface of the substrate by using a chemical coupling procedure and an ink jet application apparatus, as described in PCT application WO95/251116 (Baldeschweiler et al.)","which is incorporated herein in its entirety by reference.","In another aspect, a “gridded” array analogous to a dot (or slot) blot may be used to arrange and link cDNA fragments or oligonucleotides to the surface of a substrate using a vacuum system, thermal, UV, mechanical or chemical bonding procedures.","An array, such as those described above, may be produced by hand or by using available devices (slot blot or dot blot apparatus), materials (any suitable solid support), and machines (including robotic instruments), and may contain 8, 24, 96, 384, 1536, 6144 or more oligonucleotides, or any other number between two and one million which lends itself to the efficient use of commercially available instrumentation.","In order to conduct sample analysis using a microarray, the RNA or DNA from a biological sample is made into hybridization probes.","The mRNA is isolated, and cDNA is produced and used as a template to make antisense RNA (aRNA).","The aRNA is amplified in the presence of fluorescent nucleotides, and labeled probes are incubated with the microarray so that the probe sequences hybridize to complementary oligonucleotides of the microarray.","Incubation conditions are adjusted so that hybridization occurs with precise complementary matches or with various degrees of less complementarity.","After removal of nonhybridized probes, a scanner is used to determine the levels and patterns of fluorescence.","The scanned images are examined to determine degree of complementarity and the relative abundance of each oligonucleotide sequence on the microarray.","The biological samples may be obtained from any bodily fluids (such as blood, urine, saliva, phlegm, gastric juices, etc.","), cultured cells, biopsies, or other tissue preparations.","A detection system may be used to measure the absence, presence, and amount of hybridization for all of the distinct sequences simultaneously.","This data may be used for large-scale correlation studies on the sequences, expression patterns, mutations, variants, or polymorphisms among samples.","Using such arrays, the present invention provides methods to identify the expression of one or more of the proteins/peptides of the present invention.","In detail, such methods comprise incubating a test sample with one or more nucleic acid molecules and assaying for binding of the nucleic acid molecule with components within the test sample.","Such assays will typically involve arrays comprising many genes, at least one of which is a gene of the present invention.","FIG.","3 provides information on SNPs that have been found in the gene encoding the Ras-like protein of the present invention.","SNPs were identified at 37 different nucleotide positions.","Some of these SNPs that are located outside the ORF and in introns may affect gene transcription.","Conditions for incubating a nucleic acid molecule with a test sample vary.","Incubation conditions depend on the format employed in the assay, the detection methods employed, and the type and nature of the nucleic acid molecule used in the assay.","One skilled in the art will recognize that any one of the commonly available hybridization, amplification or array assay formats can readily be adapted to employ the novel fragments of the human genome disclosed herein.","Examples of such assays can be found in Chard, T, An Introduction to Radioimmunoassay and Related Techniques, Elsevier Science Publishers, Amsterdam, The Netherlands (1986); Bullock, G. R. et al., Techniques in Immunocytochemistry, Academic Press, Orlando, Fla. Vol.","1 (1982), Vol.","2 (1983), Vol.","3 (1985); Tijssen, P., Practice and Theory of Enzyme Immunoassays: Laboratory Techniques in Biochemistry and Molecular Biology, Elsevier Science Publishers, Amsterdam, The Netherlands (1985).","The test samples of the present invention include cells, protein or membrane extracts of cells.","The test sample used in the above-described method will vary based on the assay format, nature of the detection method and the tissues, cells or extracts used as the sample to be assayed.","Methods for preparing nucleic acid extracts or of cells are well known in the art and can be readily be adapted in order to obtain a sample that is compatible with the system utilized.","In another embodiment of the present invention, kits are provided which contain the necessary reagents to carry out the assays of the present invention.","Specifically, the invention provides a compartmentalized kit to receive, in close confinement, one or more containers which comprises: (a) a first container comprising one of the nucleic acid molecules that can bind to a fragment of the human genome disclosed herein; and (b) one or more other containers comprising one or more of the following: wash reagents, reagents capable of detecting presence of a bound nucleic acid.","Preferred kits will include chips that are capable of detecting the expression of 10 or more, 100 or more, or 500 or more, 1000 or more, or all of the genes expressed in Human.","In detail, a compartmentalized kit includes any kit in which reagents are contained in separate containers.","Such containers include small glass containers, plastic containers, strips of plastic, glass or paper, or arraying material such as silica.","Such containers allows one to efficiently transfer reagents from one compartment to another compartment such that the samples and reagents are not cross-contaminated, and the agents or solutions of each container can be added in a quantitative fashion from one compartment to another.","Such containers will include a container which will accept the test sample, a container which contains the nucleic acid probe, containers which contain wash reagents (such as phosphate buffered saline, Tris-buffers, etc.","), and containers which contain the reagents used to detect the bound probe.","One skilled in the art will readily recognize that the previously unidentified Ras-like protein genes of the present invention can be routinely identified using the sequence information disclosed herein can be readily incorporated into one of the established kit formats which are well known in the art, particularly expression arrays.","Vectors/Host Cells The invention also provides vectors containing the nucleic acid molecules described herein.","The term “vector” refers to a vehicle, preferably a nucleic acid molecule, which can transport the nucleic acid molecules.","When the vector is a nucleic acid molecule, the nucleic acid molecules are covalently linked to the vector nucleic acid.","With this aspect of the invention, the vector includes a plasmid, single or double stranded phage, a single or double stranded RNA or DNA viral vector, or artificial chromosome, such as a BAC, PAC, YAC, OR MAC.","A vector can be maintained in the host cell as an extrachromosomal element where it replicates and produces additional copies of the nucleic acid molecules.","Alternatively, the vector may integrate into the host cell genome and produce additional copies of the nucleic acid molecules when the host cell replicates.","The invention provides vectors for the maintenance (cloning vectors) or vectors for expression (expression vectors) of the nucleic acid molecules.","The vectors can function in procaryotic or eukaryotic cells or in both (shuttle vectors).","Expression vectors contain cis-acting regulatory regions that are operably linked in the vector to the nucleic acid molecules such that transcription of the nucleic acid molecules is allowed in a host cell.","The nucleic acid molecules can be introduced into the host cell with a separate nucleic acid molecule capable of affecting transcription.","Thus, the second nucleic acid molecule may provide a trans-acting factor interacting with the cis-regulatory control region to allow transcription of the nucleic acid molecules from the vector.","Alternatively, a trans-acting factor may be supplied by the host cell.","Finally, a trans-acting factor can be produced from the vector itself.","It is understood, however, that in some embodiments, transcription and/or translation of the nucleic acid molecules can occur in a cell-free system.","The regulatory sequence to which the nucleic acid molecules described herein can be operably linked include promoters for directing mRNA transcription.","These include, but are not limited to, the left promoter from bacteriophage λ, the lac, TRP, and TAC promoters from E. coli, the early and late promoters from SV40, the CMV immediate early promoter, the adenovirus early and late promoters, and retrovirus long-terminal repeats.","In addition to control regions that promote transcription, expression vectors may also include regions that modulate transcription, such as repressor binding sites and enhancers.","Examples include the SV40 enhancer, the cytomegalovirus immediate early enhancer, polyoma enhancer, adenovirus enhancers, and retrovirus LTR enhancers.","In addition to containing sites for transcription initiation and control, expression vectors can also contain sequences necessary for transcription termination and, in the transcribed region a ribosome binding site for translation.","Other regulatory control elements for expression include initiation and termination codons as well as polyadenylation signals.","The person of ordinary skill in the art would be aware of the numerous regulatory sequences that are useful in expression vectors.","Such regulatory sequences are described, for example, in Sambrook et al., Molecular Cloning: A Laboratory Manual.","2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., (1989).","A variety of expression vectors can be used to express a nucleic acid molecule.","Such vectors include chromosomal, episomal, and virus-derived vectors, for example vectors derived from bacterial plasmids, from bacteriophage, from yeast episomes, from yeast chromosomal elements, including yeast artificial chromosomes, from viruses such as baculoviruses, papovaviruses such as SV40, Vaccinia viruses, adenoviruses, poxviruses, pseudorabies viruses, and retroviruses.","Vectors may also be derived from combinations of these sources such as those derived from plasmid and bacteriophage genetic elements, e.g.","cosmids and phagemids.","Appropriate cloning and expression vectors for prokaryotic and eukaryotic hosts are described in Sambrook et al., Molecular Cloning: A Laboratory Manual.","2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., (1989).","The regulatory sequence may provide constitutive expression in one or more host cells (i.e.","tissue specific) or may provide for inducible expression in one or more cell types such as by temperature, nutrient additive, or exogenous factor such as a hormone or other ligand.","A variety of vectors providing for constitutive and inducible expression in prokaryotic and eukaryotic hosts are well known to those of ordinary skill in the art.","The nucleic acid molecules can be inserted into the vector nucleic acid by well-known methodology.","Generally, the DNA sequence that will ultimately be expressed is joined to an expression vector by cleaving the DNA sequence and the expression vector with one or more restriction enzymes and then ligating the fragments together.","Procedures for restriction enzyme digestion and ligation are well known to those of ordinary skill in the art.","The vector containing the appropriate nucleic acid molecule can be introduced into an appropriate host cell for propagation or expression using well-known techniques.","Bacterial cells include, but are not limited to, E. coli, Streptomyces, and Salmonella typhimurium.","Eukaryotic cells include, but are not limited to, yeast, insect cells such as Drosophila, animal cells such as COS and CHO cells, and plant cells.","As described herein, it may be desirable to express the peptide as a fusion protein.","Accordingly, the invention provides fusion vectors that allow for the production of the peptides.","Fusion vectors can increase the expression of a recombinant protein, increase the solubility of the recombinant protein, and aid in the purification of the protein by acting for example as a ligand for affinity purification.","A proteolytic cleavage site may be introduced at the junction of the fusion moiety so that the desired peptide can ultimately be separated from the fusion moiety.","Proteolytic enzymes include, but are not limited to, factor Xa, thrombin, and enteroRas-like protein.","Typical fusion expression vectors include pGEX (Smith et al., Gene 67:3140 (1988)), pMAL (New England Biolabs, Beverly, Mass.)","and pRIT5 (Pharmacia, Piscataway, N.J.) which fuse glutathione S-transferase (GST), maltose E binding protein, or protein A, respectively, to the target recombinant protein.","Examples of suitable inducible non-fusion E. coli expression vectors include pTrc (Amann et al., Gene 69:301-315 (1988)>and pET 11d (Studier et al., Gene Expression Technology: Methods in Enzymology 185:60-89 (1990)).","Recombinant protein expression can be maximized in a host bacteria by providing a genetic background wherein the host cell has an impaired capacity to proteolytically cleave the recombinant protein.","(Gottesman, S., Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, Calif. (1990)119-128).","Alternatively, the sequence of the nucleic acid molecule of interest can be altered to provide preferential codon usage for a specific host cell, for example E. coli.","(Wada et al., Nucleic Acids Res.","20:2111-2118 (1992)).","The nucleic acid molecules can also be expressed by expression vectors that are operative in yeast.","Examples of vectors for expression in yeast e.g., S. cerevisiae include pYepSec1 (Baldari, et al., EMBO J.","6:229-234 (1987)), pMFa (Kurjan et al., Cell 30:933-943(1982)), pJRY88 (Schultz et al., Gene 54:113-123 (1987)), and pYES2 (Invitrogen Corporation, San Diego, Calif.).","The nucleic acid molecules can also be expressed in insect cells using, for example, baculovirus expression vectors.","Baculovirus vectors available for expression of proteins in cultured insect cells (e.g., Sf 9 cells) include the pAc series (Smith et al., Mol.","Cell Biol.","3:2156-2165 (1983)) and the pVL series (Lucklow et al., Virology 170:31-39 (1989)).","In certain embodiments of the invention, the nucleic acid molecules described herein are expressed in mammalian cells using mammalian expression vectors.","Examples of mammalian expression vectors include pCDM8 (Seed, B.","Nature 329:840(1987)) and pMT2PC (Kaufman et al., EMBO J.","6:187-195 (1987)).","The expression vectors listed herein are provided by way of example only of the well-known vectors available to those of ordinary skill in the art that would be useful to express the nucleic acid molecules.","The person of ordinary skill in the art would be aware of other vectors suitable for maintenance, propagation, or expression of the nucleic acid molecules described herein.","These are found for example in Sambrook, J., Fritsh, E. F., and Maniatis, T. Molecular Cloning: A Laboratory Manual.","2nd, ed, Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989.The invention also encompasses vectors in which the nucleic acid sequences described herein are cloned into the vector in reverse orientation, but operably linked to a regulatory sequence that permits transcription of antisense RNA.","Thus, an antisense transcript can be produced to all, or to a portion, of the nucleic acid molecule sequences described herein, including both coding and non-coding regions.","Expression of this antisense RNA is subject to each of the parameters described above in relation to expression of the sense RNA (regulatory sequences, constitutive or inducible expression, tissue-specific expression).","The invention also relates to recombinant host cells containing the vectors described herein.","Host cells therefore include prokaryotic cells, lower eukaryotic cells such as yeast, other eukaryotic cells such as insect cells, and higher eukaryotic cells such as mammalian cells.","The recombinant host cells are prepared by introducing the vector constructs described herein into the cells by techniques readily available to the person of ordinary skill in the art.","These include, but are not limited to, calcium phosphate transfection, DEAE-dextran-mediated transfection, cationic lipid-mediated transfection, electroporation, transduction, infection, lipofection, and other techniques such as those found in Sambrook, et al.","(Molecular Cloning: A Laboratory Manual.","2nd, ed, Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989).","Host cells can contain more than one vector.","Thus, different nucleotide sequences can be introduced on different vectors of the same cell.","Similarly, the nucleic acid molecules can be introduced either alone or with other nucleic acid molecules that are not related to the nucleic acid molecules such as those providing trans-acting factors for expression vectors.","When more than one vector is introduced into a cell, the vectors can be introduced independently, co-introduced, or joined to the nucleic acid molecule vector.","In the case of bacteriophage and viral vectors, these can be introduced into cells as packaged or encapsulated virus by standard procedures for infection and transduction.","Viral vectors can be replication-competent or replication-defective.","In the case in which viral replication is defective, replication will occur in host cells providing functions that complement the defects.","Vectors generally include selectable markers that enable the selection of the subpopulation of cells that contain the recombinant vector constructs.","The marker can be contained in the same vector that contains the nucleic acid molecules described herein or may be on a separate vector.","Markers include tetracycline or ampicillin-resistance genes for prokaryotic host cells and dihydrofolate reductase or neomycin resistance for eukaryotic host cells.","However, any marker that provides selection for a phenotypic trait will be effective.","While the mature proteins can be produced in bacteria, yeast, mammalian cells, and other cells under the control of the appropriate regulatory sequences, cell-free transcription and translation systems can also be used to produce these proteins using RNA derived from the DNA constructs described herein.","Where secretion of the peptide is desired, which is difficult to achieve with multi-transmembrane domain containing proteins such as kinases, appropriate secretion signals are incorporated into the vector.","The signal sequence can be endogenous to the peptides or heterologous to these peptides.","Where the peptide is not secreted into the medium, which is typically the case with kinases, the protein can be isolated from the host cell by standard disruption procedures, including freeze thaw, sonication, mechanical disruption, use of lysing agents and the like.","The peptide can then be recovered and purified by well-known purification methods including ammonium sulfate precipitation, acid extraction, anion or cationic exchange chromatography, phosphocellulose chromatography, hydrophobic-interaction chromatography, affinity chromatography, hydroxylapatite chromatography, lectin chromatography, or high performance liquid chromatography.","It is also understood that depending upon the host cell in recombinant production of the peptides described herein, the peptides can have various glycosylation patterns, depending upon the cell, or maybe non-glycosylated as when produced in bacteria.","In addition, the peptides may include an initial modified methionine in some cases as a result of a host-mediated process.","Uses of Vectors and Host Cells The recombinant host cells expressing the peptides described herein have a variety of uses.","First, the cells are useful for producing a Ras-like protein polypeptide that can be further purified to produce desired amounts of Ras-like protein or fragments.","Thus, host cells containing expression vectors are useful for peptide production.","Host cells are also useful for conducting cell-based assays involving the Ras-like protein or Ras-like protein fragments.","Thus, a recombinant host cell expressing a native Ras-like protein is useful for assaying compounds that stimulate or inhibit Ras-like protein function.","Host cells are also useful for identifying Ras-like protein mutants in which these functions are affected.","If the mutants naturally occur and give rise to a pathology, host cells containing the mutations are useful to assay compounds that have a desired effect on the mutant Ras-like protein (for example, stimulating or inhibiting function) which may not be indicated by their effect on the native Ras-like protein.","Genetically engineered host cells can be further used to produce non-human transgenic animals.","A transgenic animal is preferably a mammal, for example a rodent, such as a rat or mouse, in which one or more of the cells of the animal include a transgene.","A transgene is exogenous DNA which is integrated into the genome of a cell from which a transgenic animal develops and which remains in the genome of the mature animal in one or more cell types or tissues of the transgenic animal.","These animals are useful for studying the function of a Ras-like protein and identifying and evaluating modulators of Ras-like protein activity.","Other examples of transgenic animals include non-human primates, sheep, dogs, cows, goats, chickens, and amphibians.","A transgenic animal can be produced by introducing nucleic acid into the male pronuclei of a fertilized oocyte, e.g., by microinjection, retroviral infection, and allowing the oocyte to develop in a pseudopregnant female foster animal.","Any of the Ras-like protein nucleotide sequences can be introduced as a transgene into the genome of a non-human animal, such as a mouse.","Any of the regulatory or other sequences useful in expression vectors can form part of the transgenic sequence.","This includes intronic sequences and polyadenylation signals, if not already included.","A tissue-specific regulatory sequence(s) can be operably linked to the transgene to direct expression of the Ras-like protein to particular cells.","Methods for generating transgenic animals via embryo manipulation and microinjection, particularly animals such as mice, have become conventional in the art and are described, for example, in U.S. Pat.","Nos.","4,736,866 and 4,870,009, both by Leder et al., U.S. Pat.","No.","4,873,191 by Wagner et al.","and in Hogan, B., Manipulating the Mouse Embryo, (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1986).","Similar methods are used for production of other transgenic animals.","A transgenic founder animal can be identified based upon the presence of the transgene in its genome and/or expression of transgenic mRNA in tissues or cells of the animals.","A transgenic founder animal can then be used to breed additional animals carrying the transgene.","Moreover, transgenic animals carrying a transgene can further be bred to other transgenic animals carrying other transgenes.","A transgenic animal also includes animals in which the entire animal or tissues in the animal have been produced using the homologously recombinant host cells described herein.","In another embodiment, transgenic non-human animals can be produced which contain selected systems which allow for regulated expression of the transgene.","One example of such a system is the cre/loxP recombinase system of bacteriophage P1.For a description of the cre/loxP recombinase system, see, e.g., Lakso et al.","PNAS 89:6232-6236 (1992).","Another example of a recombinase system is the FLP recombinase system of S. cerevisiae (O'Gorman et al.","Science 251:1351-1355 (1991).","If a cre/loxP recombinase system is used to regulate expression of the transgene, animals containing transgenes encoding both the Cre recombinase and a selected protein is required.","Such animals can be provided through the construction of “double” transgenic animals, e.g., by mating two transgenic animals, one containing a transgene encoding a selected protein and the other containing a transgene encoding a recombinase.","Clones of the non-human transgenic animals described herein can also be produced according to the methods described in Wilmut, I. et al.","Nature 385:810-813 (1997) and PCT International Publication Nos.","WO 97/07668 and WO 97/07669.In brief, a cell, e.g., a somatic cell, from the transgenic animal can be isolated and induced to exit the growth cycle and enter Go phase.","The quiescent cell can then be fused, e.g., through the use of electrical pulses, to an enucleated oocyte from an animal of the same species from which the quiescent cell is isolated.","The reconstructed oocyte is then cultured such that it develops to morula or blastocyst and then transferred to pseudopregnant female foster animal.","The offspring born of this female foster animal will be a clone of the animal from which the cell, e.g., the somatic cell, is isolated.","Transgenic animals containing recombinant cells that express the peptides described herein are useful to conduct the assays described herein in an in vivo context.","Accordingly, the various physiological factors that are present in vivo and that could effect ligand binding, Ras-like protein activation, and signal transduction, may not be evident from in vitro cell-free or cell-based assays.","Accordingly, it is useful to provide non-human transgenic animals to assay in vivo Ras-like protein function, including ligand interaction, the effect of specific mutant Ras-like proteins on Ras-like protein function and ligand interaction, and the effect of chimeric Ras-like proteins.","It is also possible to assess the effect of null mutations, which is mutations that substantially or completely eliminate one or more Ras-like protein functions.","All publications and patents mentioned in the above specification are herein incorporated by reference.","Various modifications and variations of the described method and system of the invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention.","Although the invention has been described in connection with specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments.","Indeed, various modifications of the above-described modes for carrying out the invention, which are obvious to those skilled in the field of molecular biology or related fields, are intended to be within the scope of the following claims."]],"string":"[\n [\n \"Isolated human ras-like proteins acid molecules encoding these human ras-like proteins, and uses thereof\",\n \"The present invention provides amino acid sequences of polypeptides that are encoded by genes within the human genome, the Ras-like protein polypeptides of the present invention.\",\n \"The present invention specifically provides isolated polypeptide and nucleic acid molecules, methods of identifying orthologs and paralogs of the Ras-like protein polypeptides, and methods of identifying modulators of the Ras-like protein polypeptides.\"\n ],\n [\n \"1.An isolated polypeptide consisting of an amino acid sequence selected from the group consisting of: (a) an amino acid sequence shown in SEQ ID NO:2; (b) an amino acid sequence of an allelic variant of an amino acid sequence shown in SEQ ID NO:2, wherein said allelic variant is encoded by a nucleic acid molecule that hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; (c) an amino acid sequence of an ortholog of an amino acid sequence shown in SEQ ID NO:2, wherein said ortholog is encoded by a nucleic acid molecule that hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; and (d) a fragment of an amino acid sequence shown in SEQ ID NO:2, wherein said fragment comprises at least 10 contiguous amino acids.\",\n \"2.An isolated polypeptide comprising an amino acid sequence selected from the group consisting of: (a) an amino acid sequence shown in SEQ ID NO:2; (b) an amino acid sequence of an allelic variant of an amino acid sequence shown in SEQ ID NO:2, wherein said allelic variant is encoded by a nucleic acid molecule that hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; (c) an amino acid sequence of an ortholog of an amino acid sequence shown in SEQ ID NO:2, wherein said ortholog is encoded by a nucleic acid molecule that hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; and (d) a fragment of an amino acid sequence shown in SEQ ID NO:2, wherein said fragment comprises at least 10 contiguous amino acids.\",\n \"3.An isolated antibody that selectively binds to a polypeptide of claim 2.4.An isolated nucleic acid molecule consisting of a nucleotide sequence selected from the group consisting of: (a) a nucleotide sequence that encodes an amino acid sequence shown in SEQ ID NO:2; (b) a nucleotide sequence that encodes of an allelic variant of an amino acid sequence shown in SEQ ID NO:2, wherein said nucleotide sequence hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; (c) a nucleotide sequence that encodes an ortholog of an amino acid sequence shown in SEQ ID NO:2, wherein said nucleotide sequence hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; (d) a nucleotide sequence that encodes a fragment of an amino acid sequence shown in SEQ ID NO:2, wherein said fragment comprises at least 10 contiguous amino acids; and (e) a nucleotide sequence that is the complement of a nucleotide sequence of (a)-(d).\",\n \"5.An isolated nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of: (a) a nucleotide sequence that encodes an amino acid sequence shown in SEQ ID NO:2; (b) a nucleotide sequence that encodes of an allelic variant of an amino acid sequence shown in SEQ ID NO:2, wherein said nucleotide sequence hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; (c) a nucleotide sequence that encodes an ortholog of an amino acid sequence shown in SEQ ID NO:2, wherein said nucleotide sequence hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; (d) a nucleotide sequence that encodes a fragment of an amino acid sequence shown in SEQ ID NO:2, wherein said fragment comprises at least 10 contiguous amino acids; and (e) a nucleotide sequence that is the complement of a nucleotide sequence of (a)-(d).\",\n \"6.A gene chip comprising a nucleic acid molecule of claim 5.7.A transgenic non-human animal comprising a nucleic acid molecule of claim 5.8.A nucleic acid vector comprising a nucleic acid molecule of claim 5.9.A host cell containing the vector of claim 8.10.A method for producing any of the polypeptides of claim 1 comprising introducing a nucleotide sequence encoding any of the amino acid sequences in (a)-(d) into a host cell, and culturing the host cell under conditions in which the polypeptides are expressed from the nucleotide sequence.\",\n \"11.A method for producing any of the polypeptides of claim 2 comprising introducing a nucleotide sequence encoding any of the amino acid sequences in (a)-(d) into a host cell, and culturing the host cell under conditions in which the polypeptides are expressed from the nucleotide sequence.\",\n \"12.A method for detecting the presence of any of the polypeptides of claim 2 in a sample, said method comprising contacting said sample with a detection agent that specifically allows detection of the presence of the polypeptide in the sample and then detecting the presence of the polypeptide.\",\n \"13.A method for detecting the presence of a nucleic acid molecule of claim 5 in a sample, said method comprising contacting the sample with an oligonucleotide that hybridizes to said nucleic acid molecule under stringent conditions and determining whether the oligonucleotide binds to said nucleic acid molecule in the sample.\",\n \"14.A method for identifying a modulator of a polypeptide of claim 2, said method comprising contacting said polypeptide with an agent and determining if said agent has modulated the function or activity of said polypeptide.\",\n \"15.The method of claim 14, wherein said agent is administered to a host cell comprising an expression vector that expresses said polypeptide.\",\n \"16.A method for identifying an agent that binds to any of the polypeptides of claim 2, said method comprising contacting the polypeptide with an agent and assaying the contacted mixture to determine whether a complex is formed with the agent bound to the polypeptide.\",\n \"17.A pharmaceutical composition comprising an agent identified by the method of claim 16 and a pharmaceutically acceptable carrier therefor.\",\n \"18.A method for treating a disease or condition mediated by a human Ras-like protein, said method comprising administering to a patient a pharmaceutically effective amount of an agent identified by the method of claim 16.19.A method for identifying a modulator of the expression of a polypeptide of claim 2, said method comprising contacting a cell expressing said polypeptide with an agent, and determining if said agent has modulated the expression of said polypeptide.\",\n \"20.An isolated human Ras-like protein polypeptide having an amino acid sequence that shares at least 70% homology with an amino acid sequence shown in SEQ ID NO:2.21.A polypeptide according to claim 20 that shares at least 90 percent homology with an amino acid sequence shown in SEQ ID NO:2.22.An isolated nucleic acid molecule encoding a human Ras-like protein polypeptide, said nucleic acid molecule sharing at least 80 percent homology with a nucleic acid molecule shown in SEQ ID NOS:1 or 3.23.A nucleic acid molecule according to claim 22 that shares at least 90 percent homology with a nucleic acid molecule shown in SEQ ID NOS:1 or 3.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>Ras-like proteins, particularly members of the Ras-like GTPase subfamilies, are a major target for drug action and development.\",\n \"Accordingly, it is valuable to the field of pharmaceutical development to identify and characterize previously unknown members of this subfamily of Ras-like proteins.\",\n \"The present invention advances the state of the art by providing a previously unidentified human Ras-like proteins that have homology to members of the Ras-like GTPase subfamilies.\",\n \"Ras Protein Ras proteins are small regulatory GTP-binding proteins, or small G proteins, which belong to the Ras protein superfamily.\",\n \"They are monomeric GTPases, but their GTPase activity is very slow (less than one GTP molecule per minute).\",\n \"Ras proteins are key relays in the signal-transducing cascade induced by the binding of a ligand to specific receptors such as receptor tyrosine kinases (RTKs), since they trigger the MAP kinase cascade.\",\n \"The ligand can be a growth factor (epidermal growth factor (EGF), platelet-derived growth factor (PDGF), insulin, an interleukin (IL), granulocyte colony-stimulating factor (G-CSF), granulocyte/macrophage colony-stimulating factor (GM-CSF).\",\n \"Ras proteins contain sequences highly conserved during evolution.\",\n \"Their tertiary structure includes ten loops connecting six strands of beta-sheet and five alpha helices.\",\n \"In mammalians, there are four Ras proteins, which are encoded by Ha-ras, N-ras, Ki-rasA and Ki-rasB genes.\",\n \"They are composed of about 170 residues and have a relative molecular mass of 21 kD.\",\n \"Ras proteins contain covalently attached modified lipids allowing these proteins to bind to the plasma membrane.\",\n \"Ha-Ras has a C-terminal farnesyl group, a C-terminal palmitoyl group and a N-terminal myristoyl group.\",\n \"In Ki-Ras(B), a C-terminal polylysine domain replaces the palmitoyl group.\",\n \"Ras proteins alternate between an inactive form bound to GDP and an active form bound to GTP.\",\n \"Their activation results from reactions induced by a guanine nucleotide-exchange factor (GEF).\",\n \"Their inactivation results from reactions catalyzed by a GTPase-activating protein (GAP).\",\n \"When a Ras protein is activated by a GEF such as a Sos protein, the N-terminal region of a serine/threonine kinase, called “Raf protein”, can bind to Ras protein.\",\n \"The C-terminal region of the activated Raf thus formed binds to another protein, MEK, and phosphorylates it on both specific tyrosine and serine residues.\",\n \"Active MEK phosphorylates and activates, in turn, a MAP kinase (ERK1 or ERK2), which is also a serine/threonine kinase.\",\n \"This phosphorylation occurs on both specific tyrosine and threonine residues of MAP kinase.\",\n \"MAP kinase phosphorylates many different proteins, especially nuclear transcription factors (TFs) that regulate expression of many genes during cell proliferation and differentiation.\",\n \"Recent researches suggest that, in mammalians, phosphatidyl inositol 3′-kinase (PI3-kinase) might be a target of Ras protein, instead of Raf protein.\",\n \"In certain mutations, the translation of ras genes may produce oncogenic Ras proteins.\",\n \"Ras-Like Protein Guanine nucleotide-binding proteins (GTP-binding proteins, or G proteins) participate in a wide range of regulatory functions including metabolism, growth, differentiation, signal transduction, cytoskeletal organization, and intracellular vesicle transport and secretion.\",\n \"These proteins control diverse sets of regulatory pathways in response to hormones, growth factors, neuromodulators, or other signaling molecules.\",\n \"When these molecules bind to transmembrane receptors, signals are propagated to effector molecules by intracellular signal transducing proteins.\",\n \"Many of these signal-transducing proteins are members of the Ras superfamily.\",\n \"The Ras superfamily is a class of low molecular weight (LMW) GTP-binding proteins that consist of 21-30 kDa polypeptides.\",\n \"These proteins regulate cell growth, cell cycle control, protein secretion, and intracellular vesicle interaction.\",\n \"In particular, the LMW GTP-binding proteins activate cellular proteins by transducing mitogenic signals involved in various cell functions in response to extracellular signals from receptors (Tavitian, A.\",\n \"(1995) C. R. Seances Soc.\",\n \"Biol.\",\n \"Fil.\",\n \"189:7-12).\",\n \"During this process, the hydrolysis of GTP acts as an energy source as well as an on-off switch for the GTPase activity of the LMW GTP-binding proteins.\",\n \"The Ras superfamily is comprised of five subfamilies: Ras, Rho, Ran, Rab, and ADP-ribosylation factor (ARF).\",\n \"Specifically, Ras genes are essential in the control of cell proliferation.\",\n \"Mutations in Ras genes have been associated with cancer.\",\n \"Rho proteins control signal transduction in the process of linking receptors of growth factors to actin polymerization that is necessary for cell division.\",\n \"Rab proteins control the translocation of vesicles to and from membranes for protein localization, protein processing, and secretion.\",\n \"Ran proteins are localized to the cell nucleus and play a key role in nuclear protein import, control of DNA synthesis, and cell-cycle progression.\",\n \"ARF and ARF-like proteins participate in a wide variety of cellular functions including vesicle trafficking, exocrine secretion, regulation of phospholipase activity, and endocytosis.\",\n \"Despite their sequence variations, all five subfamilies of the Ras superfamily share conserved structural features.\",\n \"Four conserved sequence regions (motifs I-IV) have been studied in the LMW GTP-binding proteins.\",\n \"Motif I is the most variable but has the conserved sequence, GXXXXGK.\",\n \"The lysine residue is essential in interacting with the beta.- and .gamma.-phosphates of GTP.\",\n \"Motif II, III, and IV contain highly conserved sequences of DTAGQ, NKXD, and EXSAX, respectively.\",\n \"Specifically, Motif II regulates the binding of gamma-phosphate of GTP; Motif III regulates the binding of GTP; and Motif IV regulates the guanine base of GTP.\",\n \"Most of the membrane-bound LMW GTP-binding proteins generally require a carboxy terminal isoprenyl group for membrane association and biological activity.\",\n \"The isoprenyl group is added posttranslationally through recognition of a terminal cysteine residue alone or a terminal cysteine-aliphatic amino acid-aliphatic amino acid-any amino acid (CAAX) motif.\",\n \"Additional membrane-binding energy is often provided by either internal palmitoylation or a carboxy terminal cluster of basic amino acids.\",\n \"The LMW GTP-binding proteins also have a variable effector region, located between motifs I and II, which is characterized as the interaction site for guanine nucleotide exchange factors (GEFs) or GTPase-activating proteins (GAPs).\",\n \"GEFs induce the release of GDP from the active form of the G protein, whereas GAPs interact with the inactive form by stimulating the GTPase activity of the G protein.\",\n \"The ARF subfamily has at least 15 distinct members encompassing both ARF and ARF-like proteins.\",\n \"ARF proteins identified to date exhibit high structural similarity and ADP-ribosylation enhancing activity.\",\n \"In contrast, several ARF-like proteins lack ADP-ribosylation enhancing activity and bind GTP differently.\",\n \"An example of ARF-like proteins is a rat protein, ARL184.ARL184 has been shown to have a molecular weight of 22 kDa and four functional GTP-binding sites (Icard-Liepkalns, C. et al., (1997) Eur.\",\n \"J. Biochem.\",\n \"246: 388-393).\",\n \"ARL184 is active in both the cytosol and the Golgi apparatus and is closely associated with acetylcholine release, suggesting that ARL184 is a potential regulatory protein associated with Ca.sup.2+-dependent release of acetylcholine.\",\n \"A number of Rho GTP-binding proteins have been identified in plasma membrane and cytoplasm.\",\n \"These include RhoA, B and C, and D, rhoG, rac 1 and 2, G25K-A and B, and TC10 (Hall, A. et al.\",\n \"(1993) Philos.\",\n \"Trans.\",\n \"R. Soc.\",\n \"Lond.\",\n \"(Biol.)\",\n \"340:267-271).\",\n \"All Rho proteins have a CAAX motif that binds a prenyl group and either a palmitoylation site or a basic amino acid-rich region, suggesting their role in membrane-associated functions.\",\n \"In particular, RhoD is a protein that functions in early endosome motility and distribution by inducing rearrangement of actin cytoskeleton and cell surface (Murphy, C. et al.\",\n \"(1996) Nature 384:427-432).\",\n \"During cell adhesion, the Rho proteins are essential for triggering focal complex assembly and integrin-dependent signal transduction (Hotchin, N. A. and Hall, A.\",\n \"(1995) J.\",\n \"Cell Biol.\",\n \"131:1857-1865).\",\n \"The Ras subfamily proteins already indicated supra are essential in transducing signals from receptor tyrosine kinases (RTKs) to a series of serine/threonine kinases which control cell growth and differentiation.\",\n \"Mutant Ras proteins, which bind but cannot hydrolyze GTP, are permanently activated and cause continuous cell proliferation or cancer.\",\n \"TC21, a Ras-like protein, is found to be highly expressed in a human teratocarcinoma cell line (Drivas, G. T. et al.\",\n \"(1990) Mol.\",\n \"Cell.\",\n \"Biol.\",\n \"10: 1793-1798).\",\n \"Rin and Rit are characterized as membrane-binding, Ras-like proteins without the lipid-binding CAAX motif and carboxy terminal cysteine (Lee, C.-H. J. et al.\",\n \"(1996) J. Neurosci.\",\n \"16: 6784-6794).\",\n \"Further, Rin is shown to localize in neurons and have calcium-dependant calmodulin-binding activity.\",\n \"Ras-Like GTPase Proteins The novel human protein, and encoding gene, provided by the present invention is related to the family of Ras-like GTPase proteins (also referred to as Ras-like GTP-binding proteins), which includes Rab proteins.\",\n \"The protein of the present invention is similar to Rab8 and low molecular weight (LMW) GTP-binding proteins isolated from an electrode lobe library of the marine ray Discopyge ommata (Ngsee et al., J. Biol.\",\n \"Chem.\",\n \"266 (4), 2675-2680 (1991)), some of which were determined to be homologs of the rab1, ra1, Krev, and rho LMW GTP-binding proteins.\",\n \"These proteins were localized to cholinergic synaptic vesicles and at least two of these proteins, oral and o-rho, were localized to the pre-synaptic terminals (Ngsee et al., J. Biol.\",\n \"Chem.\",\n \"266 (4), 2675-2680 (1991)).\",\n \"Rab proteins are important for regulating the targeting and fusion of membranous vesicles during organelle assembly and transport.\",\n \"Rab proteins undergo controlled exchange of GTP for GDP, and they hydrolyze GTP in a reaction that may regulate the timing and unidirectional nature of these assemblies.\",\n \"Generally, known Rab proteins terminate in sequences such as cys-X-cys (e.g., RAB3A), cys-cys (e.g., RAB1A), or a similar sequence, and generally all are geranylgeranylated.\",\n \"Rab GTP-binding proteins are similar to YPT1/SEC4 in Saccharomyces cerevisiae , which are critical for transport along the exocytic route (Chavrier et al., Mol Cell Biol 1990 December;10(12):6578-85).\",\n \"Different Rab proteins are presumed to control different steps in membrane traffic, leading to a high level of diversity and complexity within the Rab family (Chavrier et al., Mol Cell Biol 1990 December;10(12):6578-85).\",\n \"The Rab1 gene maps in close viscinity to the ‘wobbler’ spinal muscular atrophy gene.\",\n \"Rab1 and Rab2 from the snail Lymnaea stagnalis share a very high degree (95-97%) of sequence identity with mammalian Rab1 and Rab2 over the first 178-191 N-terminal amino acids; however, the C-terminal region is almost completely divergent, except for the final 24 amino acids at the extreme ends.\",\n \"Rab1 was found to be highly expressed in the albumin gland of Lymnaea stagnalis , suggesting an important role in that gland (Agterberg et al., Eur.\",\n \"J. Biochem.\",\n \"217 (1), 241-246 (1993)).\",\n \"The tethering factor p115 is a RAB1 effector that binds directly to activated RAB1.It is thought that RAB1-regulated assembly of functional effector-SNARE complexes serves as a conserved molecular mechanism for regulating recognition between different subcellular compartments such as endoplasmic reticulum and Golgi apparatus (Allan et al., Science 289: 444-448, 2000).\",\n \"GTPases play important roles in a wide variety of cell functions such as signal transduction, cytoskeletal organization, and membrane trafficking.\",\n \"Rab GTPases are particularly important for regulating cellular membrane dynamics by modulating the activity of effector proteins that then regulate vesicle trafficking.\",\n \"The Rab8 GTPase plays important roles in Golgi to plasma membrane vesicle trafficking.\",\n \"Studies have suggested that Rab37 plays an important role in mast cell degranulation.\",\n \"Thus, novel human Rab GTPases may be valuable as potential therapeutic targets for the development of allergy treatments (Masuda et al., FEBS Lett 2000 Mar.\",\n \"17;470).\",\n \"Rab5 may act, together with Rab3A, to regulate synaptic vesicle membrane flow within nerve terminals, thereby regulating neurotransmitter release.\",\n \"Rab15 and Rab3A are low molecular weight GTP-binding proteins.\",\n \"Rab proteins are generally comprised of four conserved structural domains necessary for GTP binding, as well as additional domains for membrane localization and effector protein interactions.\",\n \"Rab15 is expressed primarily in neural tissues such as the brain and is localized to synaptic vesicles (Elferink et al., J. Biol.\",\n \"Chem.\",\n \"267 (9), 5768-5775 (1992)).\",\n \"For a further review of Rab proteins, see Wedemeyer et al., Genomics 32: 447-454, 1996 and Zahraoui et al., J. Biol.\",\n \"Chem.\",\n \"264: 12394-12401, 1989.Due to their importance in human physiology, particularly in regulating membrane trafficking, novel human Ras-like GTPase proteins/genes, such as provided by the present invention, are valuable as potential targets for the development of therapeutics to treat a wide variety of diseases/disorders caused or influenced by defects in membrane trafficking.\",\n \"Furthermore, SNPs in Ras-like GTPase genes are valuable markers for the diagnosis, prognosis, prevention, and/or treatment of such diseases/disorders.\",\n \"Using the information provided by the present invention, reagents such as probes/primers for detecting the SNPs or the expression of the protein/gene provided herein may be readily developed and, if desired, incorporated into kit formats such as nucleic acid arrays, primer extension reactions coupled with mass spec detection (for SNP detection), or TaqMan PCR assays (Applied Biosystems, Foster City, Calif.).\",\n \"The discovery of new human Ras-like proteins and the polynucleotides that encode them satisfies a need in the art by providing new compositions that are useful in the diagnosis, prevention, and treatment of inflammation and disorders associated with cell proliferation and apoptosis.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>The present invention is based in part on the identification of amino acid sequences of human Ras-like protein polypeptides and proteins that are related to the Ras-like GTPase protein subfamily, as well as allelic variants and other mammalian orthologs thereof.\",\n \"These unique peptide sequences, and nucleic acid sequences that encode these peptides, can be used as models for the development of human therapeutic targets, aid in the identification of therapeutic proteins, and serve as targets for the development of human therapeutic agents that modulate Ras-like protein activity in cells and tissues that express the Ras-like protein.\",\n \"Experimental data as provided in FIG.\",\n \"1 indicates expression in bone marrow, stem cells, and leukocytes.\"\n ],\n [\n \"FIELD OF THE INVENTION The present invention is in the field of Ras-like proteins that are related to the Ras-like GTPase subfamily, recombinant DNA molecules and protein production.\",\n \"The present invention specifically provides novel Ras-like protein polypeptides and proteins and nucleic acid molecules encoding such peptide and protein molecules, all of which are useful in the development of human therapeutics and diagnostic compositions and methods.\",\n \"BACKGROUND OF THE INVENTION Ras-like proteins, particularly members of the Ras-like GTPase subfamilies, are a major target for drug action and development.\",\n \"Accordingly, it is valuable to the field of pharmaceutical development to identify and characterize previously unknown members of this subfamily of Ras-like proteins.\",\n \"The present invention advances the state of the art by providing a previously unidentified human Ras-like proteins that have homology to members of the Ras-like GTPase subfamilies.\",\n \"Ras Protein Ras proteins are small regulatory GTP-binding proteins, or small G proteins, which belong to the Ras protein superfamily.\",\n \"They are monomeric GTPases, but their GTPase activity is very slow (less than one GTP molecule per minute).\",\n \"Ras proteins are key relays in the signal-transducing cascade induced by the binding of a ligand to specific receptors such as receptor tyrosine kinases (RTKs), since they trigger the MAP kinase cascade.\",\n \"The ligand can be a growth factor (epidermal growth factor (EGF), platelet-derived growth factor (PDGF), insulin, an interleukin (IL), granulocyte colony-stimulating factor (G-CSF), granulocyte/macrophage colony-stimulating factor (GM-CSF).\",\n \"Ras proteins contain sequences highly conserved during evolution.\",\n \"Their tertiary structure includes ten loops connecting six strands of beta-sheet and five alpha helices.\",\n \"In mammalians, there are four Ras proteins, which are encoded by Ha-ras, N-ras, Ki-rasA and Ki-rasB genes.\",\n \"They are composed of about 170 residues and have a relative molecular mass of 21 kD.\",\n \"Ras proteins contain covalently attached modified lipids allowing these proteins to bind to the plasma membrane.\",\n \"Ha-Ras has a C-terminal farnesyl group, a C-terminal palmitoyl group and a N-terminal myristoyl group.\",\n \"In Ki-Ras(B), a C-terminal polylysine domain replaces the palmitoyl group.\",\n \"Ras proteins alternate between an inactive form bound to GDP and an active form bound to GTP.\",\n \"Their activation results from reactions induced by a guanine nucleotide-exchange factor (GEF).\",\n \"Their inactivation results from reactions catalyzed by a GTPase-activating protein (GAP).\",\n \"When a Ras protein is activated by a GEF such as a Sos protein, the N-terminal region of a serine/threonine kinase, called “Raf protein”, can bind to Ras protein.\",\n \"The C-terminal region of the activated Raf thus formed binds to another protein, MEK, and phosphorylates it on both specific tyrosine and serine residues.\",\n \"Active MEK phosphorylates and activates, in turn, a MAP kinase (ERK1 or ERK2), which is also a serine/threonine kinase.\",\n \"This phosphorylation occurs on both specific tyrosine and threonine residues of MAP kinase.\",\n \"MAP kinase phosphorylates many different proteins, especially nuclear transcription factors (TFs) that regulate expression of many genes during cell proliferation and differentiation.\",\n \"Recent researches suggest that, in mammalians, phosphatidyl inositol 3′-kinase (PI3-kinase) might be a target of Ras protein, instead of Raf protein.\",\n \"In certain mutations, the translation of ras genes may produce oncogenic Ras proteins.\",\n \"Ras-Like Protein Guanine nucleotide-binding proteins (GTP-binding proteins, or G proteins) participate in a wide range of regulatory functions including metabolism, growth, differentiation, signal transduction, cytoskeletal organization, and intracellular vesicle transport and secretion.\",\n \"These proteins control diverse sets of regulatory pathways in response to hormones, growth factors, neuromodulators, or other signaling molecules.\",\n \"When these molecules bind to transmembrane receptors, signals are propagated to effector molecules by intracellular signal transducing proteins.\",\n \"Many of these signal-transducing proteins are members of the Ras superfamily.\",\n \"The Ras superfamily is a class of low molecular weight (LMW) GTP-binding proteins that consist of 21-30 kDa polypeptides.\",\n \"These proteins regulate cell growth, cell cycle control, protein secretion, and intracellular vesicle interaction.\",\n \"In particular, the LMW GTP-binding proteins activate cellular proteins by transducing mitogenic signals involved in various cell functions in response to extracellular signals from receptors (Tavitian, A.\",\n \"(1995) C. R. Seances Soc.\",\n \"Biol.\",\n \"Fil.\",\n \"189:7-12).\",\n \"During this process, the hydrolysis of GTP acts as an energy source as well as an on-off switch for the GTPase activity of the LMW GTP-binding proteins.\",\n \"The Ras superfamily is comprised of five subfamilies: Ras, Rho, Ran, Rab, and ADP-ribosylation factor (ARF).\",\n \"Specifically, Ras genes are essential in the control of cell proliferation.\",\n \"Mutations in Ras genes have been associated with cancer.\",\n \"Rho proteins control signal transduction in the process of linking receptors of growth factors to actin polymerization that is necessary for cell division.\",\n \"Rab proteins control the translocation of vesicles to and from membranes for protein localization, protein processing, and secretion.\",\n \"Ran proteins are localized to the cell nucleus and play a key role in nuclear protein import, control of DNA synthesis, and cell-cycle progression.\",\n \"ARF and ARF-like proteins participate in a wide variety of cellular functions including vesicle trafficking, exocrine secretion, regulation of phospholipase activity, and endocytosis.\",\n \"Despite their sequence variations, all five subfamilies of the Ras superfamily share conserved structural features.\",\n \"Four conserved sequence regions (motifs I-IV) have been studied in the LMW GTP-binding proteins.\",\n \"Motif I is the most variable but has the conserved sequence, GXXXXGK.\",\n \"The lysine residue is essential in interacting with the beta.- and .gamma.-phosphates of GTP.\",\n \"Motif II, III, and IV contain highly conserved sequences of DTAGQ, NKXD, and EXSAX, respectively.\",\n \"Specifically, Motif II regulates the binding of gamma-phosphate of GTP; Motif III regulates the binding of GTP; and Motif IV regulates the guanine base of GTP.\",\n \"Most of the membrane-bound LMW GTP-binding proteins generally require a carboxy terminal isoprenyl group for membrane association and biological activity.\",\n \"The isoprenyl group is added posttranslationally through recognition of a terminal cysteine residue alone or a terminal cysteine-aliphatic amino acid-aliphatic amino acid-any amino acid (CAAX) motif.\",\n \"Additional membrane-binding energy is often provided by either internal palmitoylation or a carboxy terminal cluster of basic amino acids.\",\n \"The LMW GTP-binding proteins also have a variable effector region, located between motifs I and II, which is characterized as the interaction site for guanine nucleotide exchange factors (GEFs) or GTPase-activating proteins (GAPs).\",\n \"GEFs induce the release of GDP from the active form of the G protein, whereas GAPs interact with the inactive form by stimulating the GTPase activity of the G protein.\",\n \"The ARF subfamily has at least 15 distinct members encompassing both ARF and ARF-like proteins.\",\n \"ARF proteins identified to date exhibit high structural similarity and ADP-ribosylation enhancing activity.\",\n \"In contrast, several ARF-like proteins lack ADP-ribosylation enhancing activity and bind GTP differently.\",\n \"An example of ARF-like proteins is a rat protein, ARL184.ARL184 has been shown to have a molecular weight of 22 kDa and four functional GTP-binding sites (Icard-Liepkalns, C. et al., (1997) Eur.\",\n \"J. Biochem.\",\n \"246: 388-393).\",\n \"ARL184 is active in both the cytosol and the Golgi apparatus and is closely associated with acetylcholine release, suggesting that ARL184 is a potential regulatory protein associated with Ca.sup.2+-dependent release of acetylcholine.\",\n \"A number of Rho GTP-binding proteins have been identified in plasma membrane and cytoplasm.\",\n \"These include RhoA, B and C, and D, rhoG, rac 1 and 2, G25K-A and B, and TC10 (Hall, A. et al.\",\n \"(1993) Philos.\",\n \"Trans.\",\n \"R. Soc.\",\n \"Lond.\",\n \"(Biol.)\",\n \"340:267-271).\",\n \"All Rho proteins have a CAAX motif that binds a prenyl group and either a palmitoylation site or a basic amino acid-rich region, suggesting their role in membrane-associated functions.\",\n \"In particular, RhoD is a protein that functions in early endosome motility and distribution by inducing rearrangement of actin cytoskeleton and cell surface (Murphy, C. et al.\",\n \"(1996) Nature 384:427-432).\",\n \"During cell adhesion, the Rho proteins are essential for triggering focal complex assembly and integrin-dependent signal transduction (Hotchin, N. A. and Hall, A.\",\n \"(1995) J.\",\n \"Cell Biol.\",\n \"131:1857-1865).\",\n \"The Ras subfamily proteins already indicated supra are essential in transducing signals from receptor tyrosine kinases (RTKs) to a series of serine/threonine kinases which control cell growth and differentiation.\",\n \"Mutant Ras proteins, which bind but cannot hydrolyze GTP, are permanently activated and cause continuous cell proliferation or cancer.\",\n \"TC21, a Ras-like protein, is found to be highly expressed in a human teratocarcinoma cell line (Drivas, G. T. et al.\",\n \"(1990) Mol.\",\n \"Cell.\",\n \"Biol.\",\n \"10: 1793-1798).\",\n \"Rin and Rit are characterized as membrane-binding, Ras-like proteins without the lipid-binding CAAX motif and carboxy terminal cysteine (Lee, C.-H. J. et al.\",\n \"(1996) J. Neurosci.\",\n \"16: 6784-6794).\",\n \"Further, Rin is shown to localize in neurons and have calcium-dependant calmodulin-binding activity.\",\n \"Ras-Like GTPase Proteins The novel human protein, and encoding gene, provided by the present invention is related to the family of Ras-like GTPase proteins (also referred to as Ras-like GTP-binding proteins), which includes Rab proteins.\",\n \"The protein of the present invention is similar to Rab8 and low molecular weight (LMW) GTP-binding proteins isolated from an electrode lobe library of the marine ray Discopyge ommata (Ngsee et al., J. Biol.\",\n \"Chem.\",\n \"266 (4), 2675-2680 (1991)), some of which were determined to be homologs of the rab1, ra1, Krev, and rho LMW GTP-binding proteins.\",\n \"These proteins were localized to cholinergic synaptic vesicles and at least two of these proteins, oral and o-rho, were localized to the pre-synaptic terminals (Ngsee et al., J. Biol.\",\n \"Chem.\",\n \"266 (4), 2675-2680 (1991)).\",\n \"Rab proteins are important for regulating the targeting and fusion of membranous vesicles during organelle assembly and transport.\",\n \"Rab proteins undergo controlled exchange of GTP for GDP, and they hydrolyze GTP in a reaction that may regulate the timing and unidirectional nature of these assemblies.\",\n \"Generally, known Rab proteins terminate in sequences such as cys-X-cys (e.g., RAB3A), cys-cys (e.g., RAB1A), or a similar sequence, and generally all are geranylgeranylated.\",\n \"Rab GTP-binding proteins are similar to YPT1/SEC4 in Saccharomyces cerevisiae, which are critical for transport along the exocytic route (Chavrier et al., Mol Cell Biol 1990 December;10(12):6578-85).\",\n \"Different Rab proteins are presumed to control different steps in membrane traffic, leading to a high level of diversity and complexity within the Rab family (Chavrier et al., Mol Cell Biol 1990 December;10(12):6578-85).\",\n \"The Rab1 gene maps in close viscinity to the ‘wobbler’ spinal muscular atrophy gene.\",\n \"Rab1 and Rab2 from the snail Lymnaea stagnalis share a very high degree (95-97%) of sequence identity with mammalian Rab1 and Rab2 over the first 178-191 N-terminal amino acids; however, the C-terminal region is almost completely divergent, except for the final 24 amino acids at the extreme ends.\",\n \"Rab1 was found to be highly expressed in the albumin gland of Lymnaea stagnalis, suggesting an important role in that gland (Agterberg et al., Eur.\",\n \"J. Biochem.\",\n \"217 (1), 241-246 (1993)).\",\n \"The tethering factor p115 is a RAB1 effector that binds directly to activated RAB1.It is thought that RAB1-regulated assembly of functional effector-SNARE complexes serves as a conserved molecular mechanism for regulating recognition between different subcellular compartments such as endoplasmic reticulum and Golgi apparatus (Allan et al., Science 289: 444-448, 2000).\",\n \"GTPases play important roles in a wide variety of cell functions such as signal transduction, cytoskeletal organization, and membrane trafficking.\",\n \"Rab GTPases are particularly important for regulating cellular membrane dynamics by modulating the activity of effector proteins that then regulate vesicle trafficking.\",\n \"The Rab8 GTPase plays important roles in Golgi to plasma membrane vesicle trafficking.\",\n \"Studies have suggested that Rab37 plays an important role in mast cell degranulation.\",\n \"Thus, novel human Rab GTPases may be valuable as potential therapeutic targets for the development of allergy treatments (Masuda et al., FEBS Lett 2000 Mar.\",\n \"17;470).\",\n \"Rab5 may act, together with Rab3A, to regulate synaptic vesicle membrane flow within nerve terminals, thereby regulating neurotransmitter release.\",\n \"Rab15 and Rab3A are low molecular weight GTP-binding proteins.\",\n \"Rab proteins are generally comprised of four conserved structural domains necessary for GTP binding, as well as additional domains for membrane localization and effector protein interactions.\",\n \"Rab15 is expressed primarily in neural tissues such as the brain and is localized to synaptic vesicles (Elferink et al., J. Biol.\",\n \"Chem.\",\n \"267 (9), 5768-5775 (1992)).\",\n \"For a further review of Rab proteins, see Wedemeyer et al., Genomics 32: 447-454, 1996 and Zahraoui et al., J. Biol.\",\n \"Chem.\",\n \"264: 12394-12401, 1989.Due to their importance in human physiology, particularly in regulating membrane trafficking, novel human Ras-like GTPase proteins/genes, such as provided by the present invention, are valuable as potential targets for the development of therapeutics to treat a wide variety of diseases/disorders caused or influenced by defects in membrane trafficking.\",\n \"Furthermore, SNPs in Ras-like GTPase genes are valuable markers for the diagnosis, prognosis, prevention, and/or treatment of such diseases/disorders.\",\n \"Using the information provided by the present invention, reagents such as probes/primers for detecting the SNPs or the expression of the protein/gene provided herein may be readily developed and, if desired, incorporated into kit formats such as nucleic acid arrays, primer extension reactions coupled with mass spec detection (for SNP detection), or TaqMan PCR assays (Applied Biosystems, Foster City, Calif.).\",\n \"The discovery of new human Ras-like proteins and the polynucleotides that encode them satisfies a need in the art by providing new compositions that are useful in the diagnosis, prevention, and treatment of inflammation and disorders associated with cell proliferation and apoptosis.\",\n \"SUMMARY OF THE INVENTION The present invention is based in part on the identification of amino acid sequences of human Ras-like protein polypeptides and proteins that are related to the Ras-like GTPase protein subfamily, as well as allelic variants and other mammalian orthologs thereof.\",\n \"These unique peptide sequences, and nucleic acid sequences that encode these peptides, can be used as models for the development of human therapeutic targets, aid in the identification of therapeutic proteins, and serve as targets for the development of human therapeutic agents that modulate Ras-like protein activity in cells and tissues that express the Ras-like protein.\",\n \"Experimental data as provided in FIG.\",\n \"1 indicates expression in bone marrow, stem cells, and leukocytes.\",\n \"DESCRIPTION OF THE FIGURE SHEETS FIG.\",\n \"1 provides the nucleotide sequence of a cDNA molecule that encodes the Ras-like protein of the present invention.\",\n \"(SEQ ID NO:1) In addition, structure and functional information is provided, such as ATG start, stop and tissue distribution, where available, that allows one to readily determine specific uses of inventions based on this molecular sequence.\",\n \"Experimental data as provided in FIG.\",\n \"1 indicates expression in bone marrow, stem cells, and leukocytes.\",\n \"FIG.\",\n \"2 provides the predicted amino acid sequence of the Ras-like protein of the present invention.\",\n \"(SEQ ID NO:2) In addition structure and functional information such as protein family, function, and modification sites is provided where available, allowing one to readily determine specific uses of inventions based on this molecular sequence.\",\n \"FIG.\",\n \"3 provides genomic sequences that span the gene encoding the Ras-like protein of the present invention.\",\n \"(SEQ ID NO:3) In addition structure and functional information, such as intron/exon structure, promoter location, etc., is provided where available, allowing one to readily determine specific uses of inventions based on this molecular sequence.\",\n \"As illustrated in FIG.\",\n \"3, SNPs were identified at 37 different nucleotide positions.\",\n \"DETAILED DESCRIPTION OF THE INVENTION General Description The present invention is based on the sequencing of the human genome.\",\n \"During the sequencing and assembly of the human genome, analysis of the sequence information revealed previously unidentified fragments of the human genome that encode peptides that share structural and/or sequence homology to protein/peptide/domains identified and characterized within the art as being a Ras-like protein or part of a Ras-like protein and are related to the Ras-like GTPase subfamily.\",\n \"Utilizing these sequences, additional genomic sequences were assembled and transcript and/or cDNA sequences were isolated and characterized.\",\n \"Based on this analysis, the present invention provides amino acid sequences of human Ras-like protein polypeptides that are related to the Ras-like GTPase subfamily, nucleic acid sequences in the form of transcript sequences, cDNA sequences and/or genomic sequences that encode these Ras-like protein polypeptide, nucleic acid variation (allelic information), tissue distribution of expression, and information about the closest art known protein/peptide/domain that has structural or sequence homology to the Ras-like protein of the present invention.\",\n \"In addition to being previously unknown, the peptides that are provided in the present invention are selected based on their ability to be used for the development of commercially important products and services.\",\n \"Specifically, the present peptides are selected based on homology and/or structural relatedness to known Ras-like proteins of the Ras-like GTPase subfamily and the expression pattern observed.\",\n \"Experimental data as provided in FIG.\",\n \"1 indicates expression in bone marrow, stem cells, and leukocytes.\",\n \"The art has clearly established the commercial importance of members of this family of proteins and proteins that have expression patterns similar to that of the present gene.\",\n \"Some of the more specific features of the peptides of the present invention, and the uses thereof, are described herein, particularly in the Background of the Invention and in the annotation provided in the Figures, and/or are known within the art for each of the known Ras-like GTPase family or subfamily of Ras-like proteins.\",\n \"Specific Embodiments Peptide Molecules The present invention provides nucleic acid sequences that encode protein molecules that have been identified as being members of the Ras-like protein family and are related to the Ras-like GTPase subfamily (protein sequences are provided in FIG.\",\n \"2, transcript/cDNA sequences are provided in FIG.\",\n \"1 and genomic sequences are provided in FIG.\",\n \"3).\",\n \"The peptide sequences provided in FIG.\",\n \"2, as well as the obvious variants described herein, particularly allelic variants as identified herein and using the information in FIG.\",\n \"3, will be referred herein as the Ras-like proteins or peptides of the present invention, Ras-like proteins or peptides, or peptides/proteins of the present invention.\",\n \"The present invention provides isolated peptide and protein molecules that consist of, consist essentially of, or comprise the amino acid sequences of the Ras-like protein polypeptide disclosed in the FIG.\",\n \"2, (encoded by the nucleic acid molecule shown in FIG.\",\n \"1, transcript/cDNA or FIG.\",\n \"3, genomic sequence), as well as all obvious variants of these peptides that are within the art to make and use.\",\n \"Some of these variants are described in detail below.\",\n \"As used herein, a peptide is said to be “isolated” or “purified” when it is substantially free of cellular material or free of chemical precursors or other chemicals.\",\n \"The peptides of the present invention can be purified to homogeneity or other degrees of purity.\",\n \"The level of purification will be based on the intended use.\",\n \"The critical feature is that the preparation allows for the desired function of the peptide, even if in the presence of considerable amounts of other components.\",\n \"In some uses, “substantially free of cellular material” includes preparations of the peptide having less than about 30% (by dry weight) other proteins (i.e., contaminating protein), less than about 20% other proteins, less than about 10% other proteins, or less than about 5% other proteins.\",\n \"When the peptide is recombinantly produced, it can also be substantially free of culture medium, i.e., culture medium represents less than about 20% of the volume of the protein preparation.\",\n \"The language “substantially free of chemical precursors or other chemicals” includes preparations of the peptide in which it is separated from chemical precursors or other chemicals that are involved in its synthesis.\",\n \"In one embodiment, the language “substantially free of chemical precursors or other chemicals” includes preparations of the Ras-like protein polypeptide having less than about 30% (by dry weight) chemical precursors or other chemicals, less than about 20% chemical precursors or other chemicals, less than about 10% chemical precursors or other chemicals, or less than about 5% chemical precursors or other chemicals.\",\n \"The isolated Ras-like protein polypeptide can be purified from cells that naturally express it, purified from cells that have been altered to express it (recombinant), or synthesized using known protein synthesis methods.\",\n \"Experimental data as provided in FIG.\",\n \"1 indicates expression in bone marrow, stem cells, and leukocytes.\",\n \"For example, a nucleic acid molecule encoding the Ras-like protein polypeptide is cloned into an expression vector, the expression vector introduced into a host cell and the protein expressed in the host cell.\",\n \"The protein can then be isolated from the cells by an appropriate purification scheme using standard protein purification techniques.\",\n \"Many of these techniques are described in detail below.\",\n \"Accordingly, the present invention provides proteins that consist of the amino acid sequences provided in FIG.\",\n \"2 (SEQ ID NO:2), for example, proteins encoded by the transcript/cDNA nucleic acid sequences shown in FIG.\",\n \"1 (SEQ ID NO:1) and the genomic sequences provided in FIG.\",\n \"3 (SEQ ID NO:3).\",\n \"The amino acid sequence of such a protein is provided in FIG.\",\n \"2.A protein consists of an amino acid sequence when the amino acid sequence is the final amino acid sequence of the protein.\",\n \"The present invention further provides proteins that consist essentially of the amino acid sequences provided in FIG.\",\n \"2 (SEQ ID NO:2), for example, proteins encoded by the transcript/cDNA nucleic acid sequences shown in FIG.\",\n \"1 (SEQ ID NO:1) and the genomic sequences provided in FIG.\",\n \"3 (SEQ ID NO:3).\",\n \"A protein consists essentially of an amino acid sequence when such an amino acid sequence is present with only a few additional amino acid residues, for example from about 1 to about 100 or so additional residues, typically from 1 to about 20 additional residues in the final protein.\",\n \"The present invention further provides proteins that comprise the amino acid sequences provided in FIG.\",\n \"2 (SEQ ID NO:2), for example, proteins encoded by the transcript/cDNA nucleic acid sequences shown in FIG.\",\n \"1 (SEQ ID NO:1) and the genomic sequences provided in FIG.\",\n \"3 (SEQ ID NO:3).\",\n \"A protein comprises an amino acid sequence when the amino acid sequence is at least part of the final amino acid sequence of the protein.\",\n \"In such a fashion, the protein can be only the peptide or have additional amino acid molecules, such as amino acid residues (contiguous encoded sequence) that are naturally associated with it or heterologous amino acid residues/peptide sequences.\",\n \"Such a protein can have a few additional amino acid residues or can comprise several hundred or more additional amino acids.\",\n \"The preferred classes of proteins that are comprised of the Ras-like protein polypeptide of the present invention are the naturally occurring mature proteins.\",\n \"A brief description of how various types of these proteins can be made/isolated is provided below.\",\n \"The Ras-like protein polypeptides of the present invention can be attached to heterologous sequences to form chimeric or fusion proteins.\",\n \"Such chimeric and fusion proteins comprise a Ras-like protein polypeptide operatively linked to a heterologous protein having an amino acid sequence not substantially homologous to the Ras-like protein polypeptide.\",\n \"“Operatively linked” indicates that the Ras-like protein polypeptide and the heterologous protein are fused in-frame.\",\n \"The heterologous protein can be fused to the N-terminus or C-terminus of the Ras-like protein polypeptide.\",\n \"In some uses, the fusion protein does not affect the activity of the Ras-like protein polypeptide per se.\",\n \"For example, the fusion protein can include, but is not limited to, enzymatic fusion proteins, for example beta-galactosidase fusions, yeast two-hybrid GAL fusions, poly-His fusions, MYC-tagged, HI-tagged and Ig fusions.\",\n \"Such fusion proteins, particularly poly-His fusions, can facilitate the purification of recombinant Ras-like protein polypeptide.\",\n \"In certain host cells (e.g., mammalian host cells), expression and/or secretion of a protein can be increased by using a heterologous signal sequence.\",\n \"A chimeric or fusion protein can be produced by standard recombinant DNA techniques.\",\n \"For example, DNA fragments coding for the different protein sequences are ligated together in-frame in accordance with conventional techniques.\",\n \"In another embodiment, the fusion gene can be synthesized by conventional techniques including automated DNA synthesizers.\",\n \"Alternatively, PCR amplification of gene fragments can be carried out using anchor primers which give rise to complementary overhangs between two consecutive gene fragments which can subsequently be annealed and re-amplified to generate a chirneric gene sequence (see Ausubel et al., Current Protocols in Molecular Biology, 1992).\",\n \"Moreover, many expression vectors are commercially available that already encode a fusion moiety (e.g., a GST protein).\",\n \"A Ras-like protein polypeptide-encoding nucleic acid can be cloned into such an expression vector such that the fusion moiety is linked in-frame to the Ras-like protein polypeptide.\",\n \"As mentioned above, the present invention also provides and enables obvious variants of the amino acid sequence of the peptides of the present invention, such as naturally occurring mature forms of the peptide, allelic/sequence variants of the peptides, non-naturally occurring recombinantly derived variants of the peptides, and orthologs and paralogs of the peptides.\",\n \"Such variants can readily be generated using art know techniques in the fields of recombinant nucleic acid technology and protein biochemistry.\",\n \"It is understood, however, that variants exclude any amino acid sequences disclosed prior to the invention.\",\n \"Such variants can readily be identified/made using molecular techniques and the sequence information disclosed herein.\",\n \"Further, such variants can readily be distinguished from other peptides based on sequence and/or structural homology to the Ras-like protein polypeptides of the present invention.\",\n \"The degree of homology/identity present will be based primarily on whether the peptide is a functional variant or non-functional variant, the amount of divergence present in the paralog family, and the evolutionary distance between the orthologs.\",\n \"To determine the percent identity of two amino acid sequences or two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second amino acid or nucleic acid sequence for optimal alignment and non-homologous sequences can be disregarded for comparison purposes).\",\n \"In a preferred embodiment, the length of a reference sequence aligned for comparison purposes is at least 30%, 40%, 50%, 60%, 70%, 80%, or 90% or more of the length of the reference sequence.\",\n \"The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared.\",\n \"When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position (as used herein amino acid or nucleic acid “identity” is equivalent to amino acid or nucleic acid “homology”).\",\n \"The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which need to be introduced for optimal alignment of the two sequences.\",\n \"The comparison of sequences and determination of percent identity and similarity between two sequences can be accomplished using a mathematical algorithm.\",\n \"(Computational Molecular Biology, Lesk, A. M., ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, D. W., ed., Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part 1, Griffin, A. M., and Griffin, H. G., eds., Humana Press, New Jersey, 1994; Sequence Analysis in Molecular Biology, von Heinj e, G., Academic Press, 1987; and Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991).\",\n \"In a preferred embodiment, the percent identity between two amino acid sequences is determined using the Needleman and Wunsch (J. Mol.\",\n \"Biol.\",\n \"(48):444-453 (1970)) algorithm which has been incorporated into the GAP program in the GCG software package (available at http://www.gcg.com), using either a Blossom 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6.In yet another preferred embodiment, the percent identity between two nucleotide sequences is determined using the GAP program in the GCG software package (Devereux, J., et al, Nucleic Acids Res.\",\n \"12(1):387 (1984)) (available at http://www.gcg.com), using a NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 80 and a length weight of 1, 2, 3, 4, 5, or 6.In another embodiment, the percent identity between two amino acid or nucleotide sequences is determined using the algorithm of E. Meyers and W. Miller (CABIOS, 4:11-17 (1989)) which has been incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4.The nucleic acid and protein sequences of the present invention can further be used as a “query sequence” to perform a search against sequence databases to, for example, identify other family members or related sequences.\",\n \"Such searches can be performed using the NBLAST and XBLAST programs (version 2.0) of Altschul, et al.\",\n \"(J. Mol.\",\n \"Biol.\",\n \"215:403-10 (1990)).\",\n \"BLAST nucleotide searches can be performed with the NBLAST program, score=100, word length=12 to obtain nucleotide sequences homologous to the nucleic acid molecules of the invention.\",\n \"BLAST protein searches can be performed with the XBLAST program, score=50, word length=3, to obtain amino acid sequences homologous to the proteins of the invention.\",\n \"To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al.\",\n \"(Nucleic Acids Res.\",\n \"25(17):3389-3402 (1997)).\",\n \"When utilizing BLAST and gapped BLAST programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used.\",\n \"See http://www.ncbi.nlm.nih.gov.\",\n \"Full-length pre-processed forms, as well as mature processed forms, of proteins that comprise one of the peptides of the present invention can readily be identified as having complete sequence identity to one of the Ras-like protein polypeptides of the present invention as well as being encoded by the same genetic locus as the Ras-like protein polypeptide provided herein.\",\n \"The gene encoding the novel Ras-like protein of the present invention is located on a genome component that has been mapped to human chromosome 6 (as indicated in FIG.\",\n \"3), which is supported by multiple lines of evidence, such as STS and BAC map data.\",\n \"Allelic variants of a Ras-like protein polypeptide can readily be identified as being a human protein having a high degree (significant) of sequence homology/identity to at least a portion of the Ras-like protein polypeptide as well as being encoded by the same genetic locus as the Ras-like protein polypeptide provided herein.\",\n \"Genetic locus can readily be determined based on the genomic information provided in FIG.\",\n \"3, such as the genomic sequence mapped to the reference human.\",\n \"The gene encoding the novel Ras-like protein of the present invention is located on a genome component that has been mapped to human chromosome 6 (as indicated in FIG.\",\n \"3), which is supported by multiple lines of evidence, such as STS and BAC map data.\",\n \"As used herein, two proteins (or a region of the proteins) have significant homology when the amino acid sequences are typically at least about 70-80%, 80-90%, and more typically at least about 90-95% or more homologous.\",\n \"A significantly homologous amino acid sequence, according to the present invention, will be encoded by a nucleic acid sequence that will hybridize to a Ras-like protein polypeptide encoding nucleic acid molecule under stringent conditions as more fully described below.\",\n \"FIG.\",\n \"3 provides information on SNPs that have been found in the gene encoding the Ras-like protein of the present invention.\",\n \"SNPs were identified at 37 different nucleotide positions.\",\n \"Some of these SNPs that are located outside the ORF and in introns may affect gene transcription.\",\n \"Paralogs of a Ras-like protein polypeptide can readily be identified as having some degree of significant sequence homology/identity to at least a portion of the Ras-like protein polypeptide, as being encoded by a gene from humans, and as having similar activity or function.\",\n \"Two proteins will typically be considered paralogs when the amino acid sequences are typically at least about 40-50%, 50-60%, and more typically at least about 60-70% or more homologous through a given region or domain.\",\n \"Such paralogs will be encoded by a nucleic acid sequence that will hybridize to a Ras-like protein polypeptide encoding nucleic acid molecule under moderate to stringent conditions as more fully described below.\",\n \"Orthologs of a Ras-like protein polypeptide can readily be identified as having some degree of significant sequence homology/identity to at least a portion of the Ras-like protein polypeptide as well as being encoded by a gene from another organism.\",\n \"Preferred orthologs will be isolated from mammals, preferably primates, for the development of human therapeutic targets and agents.\",\n \"Such orthologs will be encoded by a nucleic acid sequence that will hybridize to a Ras-like protein polypeptide encoding nucleic acid molecule under moderate to stringent conditions, as more fully described below, depending on the degree of relatedness of the two organisms yielding the proteins.\",\n \"Non-naturally occurring variants of the Ras-like protein polypeptides of the present invention can readily be generated using recombinant techniques.\",\n \"Such variants include, but are not limited to deletions, additions and substitutions in the amino acid sequence of the Ras-like protein polypeptide.\",\n \"For example, one class of substitutions is conserved amino acid substitutions.\",\n \"Such substitutions are those that substitute a given amino acid in a Ras-like protein polypeptide by another amino acid of like characteristics.\",\n \"Typically seen as conservative substitutions are the replacements, one for another, among the aliphatic amino acids Ala, Val, Leu, and Ile; interchange of the hydroxyl residues Ser and Thr, exchange of the acidic residues Asp and Glu, substitution between the amide residues Asn and Gln, exchange of the basic residues Lys and Arg, replacements among the aromatic residues Phe, Tyr, and the like.\",\n \"Guidance concerning which amino acid changes are likely to be phenotypically silent are found in Bowie et al., Science 247:1306-1310 (1990).\",\n \"Variant Ras-like protein polypeptides can be fully functional or can lack function in one or more activities.\",\n \"Fully functional variants typically contain only conservative variations or variations in non-critical residues or in non-critical regions.\",\n \"Functional variants can also contain substitution of similar amino acids that result in no change or an insignificant change in function.\",\n \"Alternatively, such substitutions may positively or negatively affect function to some degree.\",\n \"Non-functional variants typically contain one or more non-conservative amino acid substitutions, deletions, insertions, inversions, or truncation or a substitution, insertion, inversion, or deletion in a critical residue or critical region.\",\n \"Amino acids that are essential for function can be identified by methods known in the art, such as site-directed mutagenesis or alanine-scanning mutagenesis (Cunningham et al., Science 244:1081-1085 (1989)).\",\n \"The latter procedure introduces single alanine mutations at every residue in the molecule.\",\n \"The resulting mutant molecules are then tested for biological activity such as receptor binding or in vitro proliferative activity.\",\n \"Sites that are critical for ligand-receptor binding can also be determined by structural analysis such as crystallography, nuclear magnetic resonance, or photoaffinity labeling (Smith et al., J. Mol.\",\n \"Biol.\",\n \"224:899-904 (1992); de Vos et al.\",\n \"Science 255:306-312 (1992)).\",\n \"The present invention further provides fragments of the Ras-like protein polypeptides, in addition to proteins and peptides that comprise and consist of such fragments.\",\n \"Particularly those comprising the residues identified in FIG.\",\n \"2.The fragments to which the invention pertains, however, are not to be construed as encompassing fragments that have been disclosed publicly prior to the present invention.\",\n \"As used herein, a fragment comprises at least 8, 10, 12, 14, 16 or more contiguous amino acid residues from a Ras-like protein polypeptide.\",\n \"Such fragments can be chosen based on the ability to retain one or more of the biological activities of the Ras-like protein polypeptide, or can be chosen for the ability to perform a function, e.g., act as an immunogen.\",\n \"Particularly important fragments are biologically active fragments, peptides that are, for example about 8 or more amino acids in length.\",\n \"Such fragments will typically comprise a domain or motif of the Ras-like protein polypeptide, e.g., active site.\",\n \"Further, possible fragments include, but are not limited to, domain or motif containing fragments, soluble peptide fragments, and fragments containing immunogenic structures.\",\n \"Predicted domains and functional sites are readily identifiable by computer programs well known and readily available to those of skill in the art (e.g., PROSITE, HMMer, eMOTIF, etc.).\",\n \"The results of one such analysis are provided in FIG.\",\n \"2.Polypeptides often contain amino acids other than the 20 amino acids commonly referred to as the 20 naturally occurring amino acids.\",\n \"Further, many amino acids, including the terminal amino acids, may be modified by natural processes, such as processing and other post-translational modifications, or by chemical modification techniques well known in the art.\",\n \"Common modifications that occur naturally in Ras-like protein polypeptides are described in basic texts, detailed monographs, and the research literature, and they are well known to those of skill in the art (some of these features are identified in FIG.\",\n \"2).\",\n \"Known modifications include, but are not limited to, acetylation, acylation, ADP-ribosylation, amidation, covalent attachment of flavin, covalent attachment of a heme moiety, covalent attachment of a nucleotide or nucleotide derivative, covalent attachment of a lipid or lipid derivative, covalent attachment of phosphotidylinositol, cross-linking, cyclization, disulfide bond formation, demethylation, formation of covalent crosslinks, formation of cystine, formation of pyroglutamate, formylation, gamma carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, myristoylation, oxidation, proteolytic processing, phosphorylation, prenylation, racemization, selenoylation, sulfation, transfer-RNA mediated addition of amino acids to proteins such as arginylation, and ubiquitination.\",\n \"Such modifications are well known to those of skill in the art and have been described in great detail in the scientific literature.\",\n \"Several particularly common modifications, glycosylation, lipid attachment, sulfation, gamma-carboxylation of glutamic acid residues, hydroxylation and ADP-ribosylation, for instance, are described in most basic texts, such as Proteins—Structure and Molecular Properties, 2nd Ed., T. E. Creighton, W. H. Freeman and Company, New York (1993).\",\n \"Many detailed reviews are available on this subject, such as by Wold, F., Posttranslational Covalent Modification of proteins, B. C. Johnson, Ed., Academic Press, New York 1-12 (1983); Seifter et al.\",\n \"(Meth.\",\n \"Enzymol.\",\n \"182: 626-646 (1990)) and Rattan et al.\",\n \"(Ann.\",\n \"N.Y. Acad.\",\n \"Sci.\",\n \"663:48-62 (1992)).\",\n \"Accordingly, the Ras-like protein polypeptides of the present invention also encompass derivatives or analogs in which a substituted amino acid residue is not one encoded by the genetic code, in which a substituent group is included, in which the mature Ras-like protein polypeptide is fused with another compound, such as a compound to increase the half-life of the Ras-like protein polypeptide (for example, polyethylene glycol), or in which the additional amino acids are fused to the mature Ras-like protein polypeptide, such as a leader or secretory sequence or a sequence for purification of the mature Ras-like protein polypeptide, or a pro-protein sequence.\",\n \"Protein/Peptide Uses The proteins of the present invention can be used in assays to determine the biological activity of the protein, including in a panel of multiple proteins for high-throughput screening; to raise antibodies or to elicit another immune response; as a reagent (including the labeled reagent) in assays designed to quantitatively determine levels of the protein (or its ligand or receptor) in biological fluids; and as markers for tissues in which the corresponding protein is preferentially expressed (either constitutively or at a particular stage of tissue differentiation or development or in a disease state).\",\n \"Where the protein binds or potentially binds to another protein (such as, for example, in a receptor-ligand interaction), the protein can be used to identify the binding partner so as to develop a system to identify inhibitors of the binding interaction.\",\n \"Any or all of these research utilities are capable of being developed into reagent grade or kit format for commercialization as research products.\",\n \"Methods for performing the uses listed above are well known to those skilled in the art.\",\n \"References disclosing such methods include “Molecular Cloning: A Laboratory Manual”, 2d ed., Cold Spring Harbor Laboratory Press, Sambrook, J., E. F. Fritsch and T. Maniatis eds., 1989, and “Methods in Enzymology: Guide to Molecular Cloning Techniques”, Academic Press, Berger, S. L. and A. R. Kimmel eds., 1987.The potential uses of the peptides of the present invention are based primarily on the source of the protein as well as the class/action of the protein.\",\n \"For example, Ras-like proteins isolated from humans and their human/mammalian orthologs serve as targets for identifying agents for use in mammalian therapeutic applications, e.g.\",\n \"a human drug, particularly in modulating a biological or pathological response in a cell or tissue that expresses the Ras-like protein.\",\n \"Experimental data as provided in FIG.\",\n \"1 indicates that Ras-like proteins of the present invention are expressed in bone marrow and stem cells (as indicated by virtual northern blot analysis), and leukocytes (as indicated by PCR-based tissue screening panels).\",\n \"A large percentage of pharmaceutical agents are being developed that modulate the activity of Ras-like proteins, particularly members of the Ras-like GTPase subfamily (see Background of the Invention).\",\n \"The structural and functional information provided in the Background and Figures provide specific and substantial uses for the molecules of the present invention, particularly in combination with the expression information provided in FIG.\",\n \"1.Experimental data as provided in FIG.\",\n \"1 indicates expression in bone marrow, stem cells, and leukocytes.\",\n \"Such uses can readily be determined using the information provided herein, that which is known in the art, and routine experimentation.\",\n \"The proteins of the present invention (including variants and fragments that may have been disclosed prior to the present invention) are useful for biological assays related to Ras-like proteins that are related to members of the Ras-like GTPase subfamily.\",\n \"Such assays involve any of the known Ras-like protein functions or activities or properties useful for diagnosis and treatment of Ras-like protein-related conditions that are specific for the subfamily of Ras-like proteins that the one of the present invention belongs to, particularly in cells and tissues that express the Ras-like protein.\",\n \"Experimental data as provided in FIG.\",\n \"1 indicates that Ras-like proteins of the present invention are expressed in bone marrow and stem cells (as indicated by virtual northern blot analysis), and leukocytes (as indicated by PCR-based tissue screening panels).\",\n \"The proteins of the present invention are also useful in drug screening assays, in cell-based or cell-free systems.\",\n \"Cell-based systems can be native, i.e., cells that normally express the Ras-like protein, as a biopsy or expanded in cell culture.\",\n \"Experimental data as provided in FIG.\",\n \"1 indicates expression in bone marrow, stem cells, and leukocytes.\",\n \"In an alternate embodiment, cell-based assays involve recombinant host cells expressing the Ras-like protein.\",\n \"The polypeptides can be used to identify compounds that modulate Ras-like protein activity.\",\n \"Both the Ras-like protein of the present invention and appropriate variants and fragments can be used in high-throughput screens to assay candidate compounds for the ability to bind to the Ras-like protein.\",\n \"These compounds can be further screened against a functional Ras-like protein to determine the effect of the compound on the Ras-like protein activity.\",\n \"Further, these compounds can be tested in animal or invertebrate systems to determine activity/effectiveness.\",\n \"Compounds can be identified that activate (agonist) or inactivate (antagonist) the Ras-like protein to a desired degree.\",\n \"Therefore, in one embodiment, Ras-like GTPase or a fragment or derivative thereof may be administered to a subject to prevent or treat a disorder associated with an increase in apoptosis.\",\n \"Such disorders include, but are not limited to, AIDS and other infectious or genetic immunodeficiencies, neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, retinitis pigmentosa, and cerebellar degeneration, myelodysplastic syndromes such as aplastic anemia, ischemic injuries such as myocardial infarction, stroke, and reperfusion injury, toxin-induced diseases such as alcohol-induced liver damage, cirrhosis, and lathyrism, wasting diseases such as cachexia, viral infections such as those caused by hepatitis B and C, and osteoporosis.\",\n \"In another embodiment, a pharmaceutical composition comprising Ras-like GTPase may be administered to a subject to prevent or treat a disorder associated with increased apoptosis including, but not limited to, those listed above.\",\n \"In still another embodiment, an agonist which is specific for Ras-like GTPase may be administered to prevent or treat a disorder associated with increased apoptosis including, but not limited to, those listed above.\",\n \"In a further embodiment, a vector capable of expressing Ras-like GTPase, or a fragment or a derivative thereof, may be used to prevent or treat a disorder associated with increased apoptosis including, but not limited to, those listed above.\",\n \"In cancer, where Ras-like GTPase promotes cell proliferation, it is desirable to decrease its activity.\",\n \"Therefore, in one embodiment, an antagonist of Ras-like GTPase may be administered to a subject to prevent or treat cancer including, but not limited to, adenocarcinoma, leukemia, lymphoma, melanoma, myeloma, sarcoma, and teratocarcinoma, and, in particular, cancers of the adrenal gland, bladder, bone, bone marrow, brain, breast, cervix, gall bladder, ganglia, gastrointestinal tract, heart, kidney, liver, lung, muscle, ovary, pancreas, parathyroid, penis, prostate, salivary glands, skin, spleen, testis, thymus, thyroid, and uterus.\",\n \"In one aspect, an antibody specific for Ras-like GTPase may be used directly as an antagonist, or indirectly as a targeting or delivery mechanism for bringing a pharmaceutical agent to cells or tissue which express Ras-like GTPase.\",\n \"In another embodiment, a vector expressing the complement of the polynucleotide encoding Ras-like GTPase may be administered to a subject to prevent or treat a cancer including, but not limited to, the types of cancer listed above.\",\n \"In inflammation, where Ras-like GTPase promotes cell proliferation, it is desirable to decrease its activity.\",\n \"Therefore, in one embodiment, an antagonist of Ras-like GTPase may be administered to a subject to prevent or treat an inflammation.\",\n \"Disorders associated with inflammation include, but are not limited to, Addison's disease, adult respiratory distress syndrome, allergies, anemia, asthma, atherosclerosis, bronchitis, cholecystitis, Crohn's disease, ulcerative colitis, atopic dermatitis, dermatomyositis, diabetes mellitus, emphysema, atrophic gastritis, glomerulonephritis, gout, Graves' disease, hypereosinophilia, irritable bowel syndrome, lupus erythematosus, multiple sclerosis, myasthenia gravis, myocardial or pericardial inflammation, osteoarthritis, osteoporosis, pancreatitis, polymyositis, rheumatoid arthritis, scleroderma, Sjogren's syndrome, and autoimmune thyroiditis; complications of cancer, hemodialysis, extracorporeal circulation; viral, bacterial, fungal, parasitic, protozoal, and helminthic infections and trauma.\",\n \"In one aspect, an antibody specific for Ras-like GTPase may be used directly as an antagonist, or indirectly as a targeting or delivery mechanism for bringing a pharmaceutical agent to cells or tissue which express Ras-like GTPase.\",\n \"Further, the Ras-like protein polypeptides can be used to screen a compound for the ability to stimulate or inhibit interaction between the Ras-like protein and a molecule that normally interacts with the Ras-like protein, e.g.\",\n \"a ligand or a component of the signal pathway that the Ras-like protein normally interacts.\",\n \"Such assays typically include the steps of combining the Ras-like protein with a candidate compound under conditions that allow the Ras-like protein, or fragment, to interact with the target molecule, and to detect the formation of a complex between the protein and the target or to detect the biochemical consequence of the interaction with the Ras-like protein and the target, such as any of the associated effects of signal transduction.\",\n \"Candidate compounds include, for example, 1) peptides such as soluble peptides, including Ig-tailed fusion peptides and members of random peptide libraries (see, e.g., Lam et al., Nature 354:82-84 (1991); Houghten et al., Nature 354:84-86 (1991)) and combinatorial chemistry-derived molecular libraries made of D- and/or L-configuration amino acids; 2) phosphopeptides (e.g., members of random and partially degenerate, directed phosphopeptide libraries, see, e.g., Songyang et al., Cell 72:767-778 (1993)); 3) antibodies (e.g., polyclonal, monoclonal, humanized, anti-idiotypic, chimeric, and single chain antibodies as well as Fab, F(ab′)2, Fab expression library fragments, and epitope-binding fragments of antibodies); and 4) small organic and inorganic molecules (e.g., molecules obtained from combinatorial and natural product libraries).\",\n \"(Hodgson, Bio/technology, 1992, Sep. 10(9);973-80).\",\n \"One candidate compound is a soluble fragment of the Ras-like protein that competes for ligand binding.\",\n \"Other candidate compounds include mutant Ras-like proteins or appropriate fragments containing mutations that affect Ras-like protein function and thus compete for ligand.\",\n \"Accordingly, a fragment that competes for ligand, for example with a higher affinity, or a fragment that binds ligand but does not allow release, is within the scope of the invention.\",\n \"The invention further includes other end point assays to identify compounds that modulate (stimulate or inhibit) Ras-like protein activity.\",\n \"The assays typically involve an assay of events in the Ras-like protein mediated signal transduction pathway that indicate Ras-like protein activity.\",\n \"Thus, the phosphorylation of a protein/ligand target, the expression of genes that are up- or down-regulated in response to the Ras-like protein dependent signal cascade can be assayed.\",\n \"In one embodiment, the regulatory region of such genes can be operably linked to a marker that is easily detectable, such as luciferase.\",\n \"Alternatively, phosphorylation of the Ras-like protein, or a Ras-like protein target, could also be measured.\",\n \"Any of the biological or biochemical functions mediated by the Ras-like protein can be used as an endpoint assay.\",\n \"These include all of the biochemical or biochemical/biological events described herein, in the references cited herein, incorporated by reference for these endpoint assay targets, and other functions known to those of ordinary skill in the art.\",\n \"Binding and/or activating compounds can also be screened by using chimeric Ras-like proteins in which any of the protein's domains, or parts thereof, can be replaced by heterologous domains or subregions.\",\n \"Accordingly, a different set of signal transduction components is available as an end-point assay for activation.\",\n \"This allows for assays to be performed in other than the specific host cell from which the Ras-like protein is derived.\",\n \"The Ras-like protein polypeptide of the present invention is also useful in competition binding assays in methods designed to discover compounds that interact with the Ras-like protein.\",\n \"Thus, a compound is exposed to a Ras-like protein polypeptide under conditions that allow the compound to bind or to otherwise interact with the polypeptide.\",\n \"Soluble Ras-like protein polypeptide is also added to the mixture.\",\n \"If the test compound interacts with the soluble Ras-like protein polypeptide, it decreases the amount of complex formed or activity from the Ras-like protein target.\",\n \"This type of assay is particularly useful in cases in which compounds are sought that interact with specific regions of the Ras-like protein.\",\n \"Thus, the soluble polypeptide that competes with the target Ras-like protein region is designed to contain peptide sequences corresponding to the region of interest.\",\n \"To perform cell free drug screening assays, it is sometimes desirable to immobilize either the Ras-like protein, or fragment, or its target molecule to facilitate separation of complexes from uncomplexed forms of one or both of the proteins, as well as to accommodate automation of the assay.\",\n \"Techniques for immobilizing proteins on matrices can be used in the drug screening assays.\",\n \"In one embodiment, a fusion protein can be provided which adds a domain that allows the protein to be bound to a matrix.\",\n \"For example, glutathione-S-transferase/15625 fusion proteins can be adsorbed onto glutathione sepharose beads (Sigma Chemical, St. Louis, Mo.)\",\n \"or glutathione derivatized microtitre plates, which are then combined with the cell lysates (e.g., 35S-labeled) and the candidate compound, and the mixture incubated under conditions conducive to complex formation (e.g., at physiological conditions for salt and pH).\",\n \"Following incubation, the beads are washed to remove any unbound label, and the matrix immobilized and radiolabel determined directly, or in the supernatant after the complexes are dissociated.\",\n \"Alternatively, the complexes can be dissociated from the matrix, separated by SDS-PAGE, and the level of Ras-like protein-binding protein found in the bead fraction quantitated from the gel using standard electrophoretic techniques.\",\n \"For example, either the polypeptide or its target molecule can be immobilized utilizing conjugation of biotin and streptavidin with techniques well known in the art.\",\n \"Alternatively, antibodies reactive with the protein but which do not interfere with binding of the protein to its target molecule can be derivatized to the wells of the plate, and the protein trapped in the wells by antibody conjugation.\",\n \"Preparations of a Ras-like protein-binding protein and a candidate compound are incubated in the Ras-like protein-presenting wells and the amount of complex trapped in the well can be quantitated.\",\n \"Methods for detecting such complexes, in addition to those described above for the GST-immobilized complexes, include immunodetection of complexes using antibodies reactive with the Ras-like protein target molecule, or which are reactive with Ras-like protein and compete with the target molecule, as well as enzyme-linked assays which rely on detecting an enzymatic activity associated with the target molecule.\",\n \"Agents that modulate one of the Ras-like proteins of the present invention can be identified using one or more of the above assays, alone or in combination.\",\n \"It is generally preferable to use a cell-based or cell free system first and then confirm activity in an animal/insect model system.\",\n \"Such model systems are well known in the art and can readily be employed in this context.\",\n \"Modulators of Ras-like protein activity identified according to these drug screening assays can be used to treat a subject with a disorder mediated by the Ras-like protein associated pathway, by treating cells that express the Ras-like protein.\",\n \"Experimental data as provided in FIG.\",\n \"1 indicates expression in bone marrow, stem cells, and leukocytes.\",\n \"These methods of treatment include the steps of administering the modulators of protein activity in a pharmaceutical composition as described herein, to a subject in need of such treatment.\",\n \"In yet another aspect of the invention, the Ras-like proteins can be used as “bait proteins” in a two-hybrid assay or three-hybrid assay (see, e.g., U.S. Pat.\",\n \"No.\",\n \"5,283,317; Zervos et al., Cell 72:223-232 (1993); Madura et al., J. Biol.\",\n \"Chem.\",\n \"268:12046-12054 (1993); Bartel et al., Biotechniques 14:920-924 (1993); Iwabuchi et al., Oncogene 8:1693-1696 (1993); and Brent WO94/10300), to identify other proteins that bind to or interact with the Ras-like protein and are involved in Ras-like protein activity.\",\n \"Such Ras-like protein-binding proteins are also likely to be involved in the propagation of signals by the Ras-like proteins or Ras-like protein targets as, for example, downstream elements of a Ras-like protein-mediated signaling pathway, e.g., a pain signaling pathway.\",\n \"Alternatively, such Ras-like protein-binding proteins are likely to be Ras-like protein inhibitors.\",\n \"The two-hybrid system is based on the modular nature of most transcription factors, which consist of separable DNA-binding and activation domains.\",\n \"Briefly, the assay utilizes two different DNA constructs.\",\n \"In one construct, the gene that codes for a Ras-like protein is fused to a gene encoding the DNA binding domain of a known transcription factor (e.g., GAL-4).\",\n \"In the other construct, a DNA sequence, from a library of DNA sequences, that encodes an unidentified protein (“prey” or “sample”) is fused to a gene that codes for the activation domain of the known transcription factor.\",\n \"If the “bait” and the “prey” proteins are able to interact, in vivo, forming a Ras-like protein-dependent complex, the DNA-binding and activation domains of the transcription factor are brought into close proximity.\",\n \"This proximity allows transcription of a reporter gene (e.g., LacZ) which is operably linked to a transcriptional regulatory site responsive to the transcription factor.\",\n \"Expression of the reporter gene can be detected and cell colonies containing the functional transcription factor can be isolated and used to obtain the cloned gene which encodes the protein which interacts with the Ras-like protein.\",\n \"This invention further pertains to novel agents identified by the above-described screening assays.\",\n \"Accordingly, it is within the scope of this invention to further use an agent identified as described herein in an appropriate animal model.\",\n \"For example, an agent identified as described herein (e.g., a Ras-like protein modulating agent, an antisense Ras-like protein nucleic acid molecule, a Ras-like protein-specific antibody, or a Ras-like protein-binding partner) can be used in an animal or insect model to determine the efficacy, toxicity, or side effects of treatment with such an agent.\",\n \"Alternatively, an agent identified as described herein can be used in an animal or insect model to determine the mechanism of action of such an agent.\",\n \"Furthermore, this invention pertains to uses of novel agents identified by the above-described screening assays for treatments as described herein.\",\n \"The Ras-like proteins of the present invention are also useful to provide a target for diagnosing a disease or predisposition to a disease mediated by the peptide, Accordingly, the invention provides methods for detecting the presence, or levels of, the protein (or encoding mRNA) in a cell, tissue, or organism.\",\n \"Experimental data as provided in FIG.\",\n \"1 indicates expression in bone marrow, stem cells, and leukocytes.\",\n \"The method involves contacting a biological sample with a compound capable of interacting with the receptor protein such that the interaction can be detected.\",\n \"Such an assay can be provided in a single detection format or a multi-detection format such as an antibody chip array.\",\n \"One agent for detecting a protein in a sample is an antibody capable of selectively binding to protein.\",\n \"A biological sample includes tissues, cells and biological fluids isolated from a subject, as well as tissues, cells, and fluids present within a subject.\",\n \"The peptides also are useful to provide a target for diagnosing a disease or predisposition to a disease mediated by the peptide, Accordingly, the invention provides methods for detecting the presence, or levels of, the protein in a cell, tissue, or organism.\",\n \"The method involves contacting a biological sample with a compound capable of interacting with the receptor protein such that the interaction can be detected.\",\n \"The peptides of the present invention also provide targets for diagnosing active disease, or predisposition to a disease, in a patient having a variant peptide.\",\n \"Thus, the peptide can be isolated from a biological sample and assayed for the presence of a genetic mutation that results in translation of an aberrant peptide.\",\n \"This includes amino acid substitution, deletion, insertion, rearrangement, (as the result of aberrant splicing events), and inappropriate post-translational modification.\",\n \"Analytic methods include altered electrophoretic mobility, altered tryptic peptide digest, altered receptor activity in cell-based or cell-free assay, alteration in ligand or antibody-binding pattern, altered isoelectric point, direct amino acid sequencing, and any other of the known assay techniques useful for detecting mutations in a protein.\",\n \"Such an assay can be provided in a single detection format or a multi-detection format such as an antibody chip array.\",\n \"In vitro techniques for detection of peptide include enzyme linked immunosorbent assays (ELISAs), Western blots, immunoprecipitations, and immunofluorescence using a detection reagents, such as an antibody or protein binding agent.\",\n \"Alternatively, the peptide can be detected in vivo in a subject by introducing into the subject a labeled anti-peptide antibody.\",\n \"For example, the antibody can be labeled with a radioactive marker whose presence and location in a subject can be detected by standard imaging techniques.\",\n \"Particularly useful are methods that detect the allelic variant of a peptide expressed in a subject and methods which detect fragments of a peptide in a sample.\",\n \"The peptides are also useful in pharmacogenomic analysis.\",\n \"Pharmacogenomics deal with clinically significant hereditary variations in the response to drugs due to altered drug disposition and abnormal action in affected persons.\",\n \"See, e.g., Eichelbaum, M. (Clin.\",\n \"Exp.\",\n \"Pharmacol.\",\n \"Physiol.\",\n \"23(10-11):983-985 (1996)), and Linder, M. W. (Clin.\",\n \"Chem.\",\n \"43(2):254-266 (1997)).\",\n \"The clinical outcomes of these variations result in severe toxicity of therapeutic drugs in certain individuals or therapeutic failure of drugs in certain individuals as a result of individual variation in metabolism.\",\n \"Thus, the genotype of the individual can determine the way a therapeutic compound acts on the body or the way the body metabolizes the compound.\",\n \"Further, the activity of drug metabolizing enzymes effects both the intensity and duration of drug action.\",\n \"Thus, the pharmacogenomics of the individual permit the selection of effective compounds and effective dosages of such compounds for prophylactic or therapeutic treatment based on the individual's genotype.\",\n \"The discovery of genetic polymorphisms in some drug metabolizing enzymes has explained why some patients do not obtain the expected drug effects, show an exaggerated drug effect, or experience serious toxicity from standard drug dosages.\",\n \"Polymorphisms can be expressed in the phenotype of the extensive metabolizer and the phenotype of the poor metabolizer.\",\n \"Accordingly, genetic polymorphism may lead to allelic protein variants of the receptor protein in which one or more of the receptor functions in one population is different from those in another population.\",\n \"The peptides thus allow a target to ascertain a genetic predisposition that can affect treatment modality.\",\n \"Thus, in a ligand-based treatment, polymorphism may give rise to amino terminal extracellular domains and/or other ligand-binding regions that are more or less active in ligand binding, and receptor activation.\",\n \"Accordingly, ligand dosage would necessarily be modified to maximize the therapeutic effect within a given population containing a polymorphism.\",\n \"As an alternative to genotyping, specific polymorphic peptides could be identified.\",\n \"The peptides are also useful for treating a disorder characterized by an absence of, inappropriate, or unwanted expression of the protein.\",\n \"Experimental data as provided in FIG.\",\n \"1 indicates expression in bone marrow, stem cells, and leukocytes.\",\n \"Accordingly, methods for treatment include the use of the Ras-like protein or fragments.\",\n \"Antibodies The invention also provides antibodies that selectively bind to one of the peptides of the present invention, a protein comprising such a peptide, as well as variants and fragments thereof.\",\n \"As used herein, an antibody selectively binds a target peptide when it binds the target peptide and does not significantly bind to unrelated proteins.\",\n \"An antibody is still considered to selectively bind a peptide even if it also binds to other proteins that are not substantially homologous with the target peptide so long as such proteins share homology with a fragment or domain of the peptide target of the antibody.\",\n \"In this case, it would be understood that antibody binding to the peptide is still selective despite some degree of cross-reactivity.\",\n \"As used herein, an antibody is defined in terrns consistent with that recognized within the art: they are multi-subunit proteins produced by a mammalian organism in response to an antigen challenge.\",\n \"The antibodies of the present invention include polyclonal antibodies and monoclonal antibodies, as well as fragments of such antibodies, including, but not limited to, Fab or F(ab′)2, and Fv fragments.\",\n \"Many methods are known for generating and/or identifying antibodies to a given target peptide.\",\n \"Several such methods are described by Harlow, Antibodies, Cold Spring Harbor Press, (1989).\",\n \"In general, to generate antibodies, an isolated peptide is used as an immunogen and is administered to a mammalian organism, such as a rat, rabbit or mouse.\",\n \"The full-length protein, an antigenic peptide fragment or a fusion protein can be used.\",\n \"Particularly important fragments are those covering functional domains, such as the domains identified in FIG.\",\n \"2, and domain of sequence homology or divergence amongst the family, such as those that can readily be identified using protein alignment methods and as presented in the Figures.\",\n \"Antibodies are preferably prepared from regions or discrete fragments of the Ras-like proteins.\",\n \"Antibodies can be prepared from any region of the peptide as described herein.\",\n \"However, preferred regions will include those involved in function/activity and/or receptor/binding partner interaction.\",\n \"FIG.\",\n \"2 can be used to identify particularly important regions while sequence alignment can be used to identify conserved and unique sequence fragments.\",\n \"An antigenic fragment will typically comprise at least 8 contiguous amino acid residues.\",\n \"The antigenic peptide can comprise, however, at least 10, 12, 14, 16 or more amino acid residues.\",\n \"Such fragments can be selected on a physical property, such as fragments correspond to regions that are located on the surface of the protein, e.g., hydrophilic regions or can be selected based on sequence uniqueness (see FIG.\",\n \"2).\",\n \"Detection of an antibody of the present invention can be facilitated by coupling (i.e., physically linking) the antibody to a detectable substance.\",\n \"Examples of detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, and radioactive materials.\",\n \"Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, β-galactosidase, or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidin/biotin and avidin/biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; an example of a luminescent material includes luminol; examples of bioluminescent materials include luciferase, luciferin, and aequorin, and examples of suitable radioactive material include 125I, 131I, 35S, or 3H.\",\n \"Antibody Uses The antibodies can be used to isolate one of the proteins of the present invention by standard techniques, such as affinity chromatography or immunoprecipitation.\",\n \"The antibodies can facilitate the purification of the natural protein from cells and recombinantly produced protein expressed in host cells.\",\n \"In addition, such antibodies are useful to detect the presence of one of the proteins of the present invention in cells or tissues to determine the pattern of expression of the protein among various tissues in an organism and over the course of normal development.\",\n \"Experimental data as provided in FIG.\",\n \"1 indicates that Ras-like proteins of the present invention are expressed in bone marrow and stem cells (as indicated by virtual northern blot analysis), and leukocytes (as indicated by PCR-based tissue screening panels).\",\n \"Further, such antibodies can be used to detect protein in situ, in vitro, or in a cell lysate or supernatant in order to evaluate the abundance and pattern of expression.\",\n \"Also, such antibodies can be used to assess abnormal tissue distribution or abnormal expression during development.\",\n \"Antibody detection of circulating fragments of the full-length protein can be used to identify turnover.\",\n \"Further, the antibodies can be used to assess expression in disease states such as in active stages of the disease or in an individual with a predisposition toward disease related to the protein's function.\",\n \"When a disorder is caused by an inappropriate tissue distribution, developmental expression, level of expression of the protein, or expressed/processed form, the antibody can be prepared against the normal protein.\",\n \"Experimental data as provided in FIG.\",\n \"1 indicates expression in bone marrow, stem cells, and leukocytes.\",\n \"If a disorder is characterized by a specific mutation in the protein, antibodies specific for this mutant protein can be used to assay for the presence of the specific mutant protein.\",\n \"The antibodies can also be used to assess normal and aberrant subcellular localization of cells in the various tissues in an organism.\",\n \"Experimental data as provided in FIG.\",\n \"1 indicates expression in bone marrow, stem cells, and leukocytes.\",\n \"The diagnostic uses can be applied, not only in genetic testing, but also in monitoring a treatment modality.\",\n \"Accordingly, where treatment is ultimately aimed at correcting expression level or the presence of aberrant sequence and aberrant tissue distribution or developmental expression, antibodies directed against the or relevant fragments can be used to monitor therapeutic efficacy.\",\n \"Additionally, antibodies are useful in pharmacogenomic analysis.\",\n \"Thus, antibodies prepared against polymorphic proteins can be used to identify individuals that require modified treatment modalities.\",\n \"The antibodies are also useful as diagnostic tools as an immunological marker for aberrant protein analyzed by electrophoretic mobility, isoelectric point, tryptic peptide digest, and other physical assays known to those in the art.\",\n \"The antibodies are also useful for tissue typing.\",\n \"Experimental data as provided in FIG.\",\n \"1 indicates expression in bone marrow, stem cells, and leukocytes.\",\n \"Thus, where a specific protein has been correlated with expression in a specific tissue, antibodies that are specific for this protein can be used to identify a tissue type.\",\n \"The antibodies are also useful for inhibiting protein function, for example, blocking the binding of the Ras-like protein to a binding partner such as a substrate.\",\n \"These uses can also be applied in a therapeutic context in which treatment involves inhibiting the protein's function.\",\n \"An antibody can be used, for example, to block binding, thus modulating (agonizing or antagonizing) the peptides activity.\",\n \"Antibodies can be prepared against specific fragments containing sites required for function or against intact protein that is associated with a cell or cell membrane.\",\n \"See FIG.\",\n \"2 for structural information relating to the proteins of the present invention.\",\n \"The invention also encompasses kits for using antibodies to detect the presence of a protein in a biological sample.\",\n \"The kit can comprise antibodies such as a labeled or labelable antibody and a compound or agent for detecting protein in a biological sample; means for determining the amount of protein in the sample; means for comparing the amount of protein in the sample with a standard; and instructions for use.\",\n \"Nucleic Acid Molecules The present invention further provides isolated nucleic acid molecules that encode a Ras-like protein polypeptide of the present invention.\",\n \"Such nucleic acid molecules will consist of, consist essentially of, or comprise a nucleotide sequence that encodes one of the Ras-like protein polypeptides of the present invention, an allelic variant thereof, or an ortholog or paralog thereof.\",\n \"As used herein, an “isolated” nucleic acid molecule is one that is separated from other nucleic acid present in the natural source of the nucleic acid.\",\n \"Preferably, an “isolated” nucleic acid is free of sequences which naturally flank the nucleic acid (i.e., sequences located at the 5′ and 3′ ends of the nucleic acid) in the genomic DNA of the organism from which the nucleic acid is derived.\",\n \"However, there can be some flanking nucleotide sequences, for example up to about 5 KB, particularly contiguous peptide encoding sequences and peptide encoding sequences within the same gene but separated by introns in the genomic sequence.\",\n \"The important point is that the nucleic acid is isolated from remote and unimportant flanking sequences such that it can be subjected to the specific manipulations described herein such as recombinant expression, preparation of probes and primers, and other uses specific to the nucleic acid sequences.\",\n \"Moreover, an “isolated” nucleic acid molecule, such as a cDNA molecule, can be substantially free of other cellular material, or culture medium when produced by recombinant techniques, or chemical precursors or other chemicals when chemically synthesized.\",\n \"However, the nucleic acid molecule can be fused to other coding or regulatory sequences and still be considered isolated.\",\n \"For example, recombinant DNA molecules contained in a vector are considered isolated.\",\n \"Further examples of isolated DNA molecules include recombinant DNA molecules maintained in heterologous host cells or purified (partially or substantially) DNA molecules in solution.\",\n \"Isolated RNA molecules include in vivo or in vitro RNA transcripts of the isolated DNA molecules of the present invention.\",\n \"Isolated nucleic acid molecules according to the present invention further include such molecules produced synthetically.\",\n \"Accordingly, the present invention provides nucleic acid molecules that consist of the nucleotide sequence shown in FIG.\",\n \"1 or 3 (SEQ ID NO:1, transcript sequence and SEQ ID NO:3, genomic sequence), or any nucleic acid molecule that encodes the protein provided in FIG.\",\n \"2, SEQ ID NO:2.A nucleic acid molecule consists of a nucleotide sequence when the nucleotide sequence is the complete nucleotide sequence of the nucleic acid molecule.\",\n \"The present invention further provides nucleic acid molecules that consist essentially of the nucleotide sequence shown in FIG.\",\n \"1 or 3 (SEQ ID NO:1, transcript sequence and SEQ ID NO:3, genomic sequence), or any nucleic acid molecule that encodes the protein provided in FIG.\",\n \"2, SEQ ID NO:2.A nucleic acid molecule consists essentially of a nucleotide sequence when such a nucleotide sequence is present with only a few additional nucleic acid residues in the final nucleic acid molecule.\",\n \"The present invention further provides nucleic acid molecules that comprise the nucleotide sequences shown in FIG.\",\n \"1 or 3 (SEQ ID NO:1, transcript sequence and SEQ ID NO:3, genomic sequence), or any nucleic acid molecule that encodes the protein provided in FIG.\",\n \"2, SEQ ID NO:2.A nucleic acid molecule comprises a nucleotide sequence when the nucleotide sequence is at least part of the final nucleotide sequence of the nucleic acid molecule.\",\n \"In such a fashion, the nucleic acid molecule can be only the nucleotide sequence or have additional nucleic acid residues, such as nucleic acid residues that are naturally associated with it or heterologous nucleotide sequences.\",\n \"Such a nucleic acid molecule can have a few additional nucleotides or can comprises several hundred or more additional nucleotides.\",\n \"A brief description of how various types of these nucleic acid molecules can be readily made/isolated is provided below.\",\n \"In FIGS.\",\n \"1 and 3, both coding and non-coding sequences are provided.\",\n \"Because of the source of the present invention, humans genomic sequence (FIG.\",\n \"3) and cDNA/transcript sequences (FIG.\",\n \"1), the nucleic acid molecules in the Figures will contain genomic intronic sequences, 5′ and 3′ non-coding sequences, gene regulatory regions and non-coding intergenic sequences.\",\n \"In general such sequence features are either noted in FIGS.\",\n \"1 and 3 or can readily be identified using computational tools known in the art.\",\n \"As discussed below, some of the non-coding regions, particularly gene regulatory elements such as promoters, are useful for a variety of purposes, e.g.\",\n \"control of heterologous gene expression, target for identifying gene activity modulating compounds, and are particularly claimed as fragments of the genomic sequence provided herein.\",\n \"Full-length genes may be cloned from known sequence using any one of a number of methods known in the art.\",\n \"For example, a method which employs XL-PCR (Perkin-Elmer, Foster City, Calif.) to amplify long pieces of DNA may be used.\",\n \"Other methods for obtaining full-length sequences are well known in the art.\",\n \"The isolated nucleic acid molecules can encode the mature protein plus additional amino or carboxyl-terminal amino acids, or amino acids interior to the mature peptide (when the mature form has more than one peptide chain, for instance).\",\n \"Such sequences may play a role in processing of a protein from precursor to a mature form, facilitate protein trafficking, prolong or shorten protein half-life, or facilitate manipulation of a protein for assay or production, among other things.\",\n \"As generally is the case in situ, the additional amino acids may be processed away from the mature protein by cellular enzymes.\",\n \"As mentioned above, the isolated nucleic acid molecules include, but are not limited to, the sequence encoding the Ras-like protein polypeptide alone, the sequence encoding the mature peptide and additional coding sequences, such as a leader or secretory sequence (e.g., a pre-pro or pro-protein sequence), the sequence encoding the mature peptide, with or without the additional coding sequences, plus additional non-coding sequences, for example introns and non-coding 5′ and 3′ sequences such as transcribed but non-translated sequences that play a role in transcription, mRNA processing (including splicing and polyadenylation signals), ribosome binding, and stability of mRNA.\",\n \"In addition, the nucleic acid molecule may be fused to a marker sequence encoding, for example, a peptide that facilitates purification.\",\n \"Isolated nucleic acid molecules can be in the form of RNA, such as mRNA, or in the form of DNA, including cDNA and genomic DNA obtained by cloning or produced by chemical synthetic techniques or by a combination thereof.\",\n \"The nucleic acid, especially DNA, can be double-stranded or single-stranded.\",\n \"Single-stranded nucleic acid can be the coding strand (sense strand) or the non-coding strand (anti-sense strand).\",\n \"The invention further provides nucleic acid molecules that encode fragments of the peptides of the present invention and that encode obvious variants of the Ras-like proteins of the present invention that are described above.\",\n \"Such nucleic acid molecules may be naturally occurring, such as allelic variants (same locus), paralogs (different locus), and orthologs (different organism), or may be constructed by recombinant DNA methods or by chemical synthesis.\",\n \"Such non-naturally occurring variants may be made by mutagenesis techniques, including those applied to nucleic acid molecules, cells, or whole organisms.\",\n \"Accordingly, as discussed above, the variants can contain nucleotide substitutions, deletions inversions, and/or insertions.\",\n \"Variation can occur in either or both the coding and non-coding regions.\",\n \"The variations can produce both conservative and non-conservative amino acid substitutions.\",\n \"The present invention further provides non-coding fragments of the nucleic acid molecules provided in the FIGS.\",\n \"1 and 3.Preferred non-coding fragments include, but are not limited to, promoter sequences, enhancer sequences, gene modulating sequences, and gene termination sequences.\",\n \"Such fragments are useful in controlling heterologous gene expression and in developing screens to identify gene-modulating agents.\",\n \"A fragment comprises a contiguous nucleotide sequence greater than 12 or more nucleotides.\",\n \"Further, a fragment could be at least 30, 40, 50, 100 250, or 500 nucleotides in length.\",\n \"The length of the fragment will be based on its intended use.\",\n \"For example, the fragment can encode epitope-bearing regions of the peptide, or can be useful as DNA probes and primers.\",\n \"Such fragments can be isolated using the known nucleotide sequence to synthesize an oligonucleotide probe.\",\n \"A labeled probe can then be used to screen a cDNA library, genomic DNA library, or mRNA to isolate nucleic acid corresponding to the coding region.\",\n \"Further, primers can be used in PCR reactions to clone specific regions of gene.\",\n \"A probe/primer typically comprises substantially a purified oligonucleotide or oligonucleotide pair.\",\n \"The oligonucleotide typically comprises a region of nucleotide sequence that hybridizes under stringent conditions to at least about 12, 20, 25, 40, 50, or more consecutive nucleotides.\",\n \"Orthologs, homologs, and allelic variants can be identified using methods well known in the art.\",\n \"As described in the Peptide Section, these variants comprise a nucleotide sequence encoding a peptide that is typically 60-70%, 70-80%, 80-90%, and more typically at least about 90-95% or more homologous to the nucleotide sequence shown in the Figure sheets or a fragment of this sequence.\",\n \"Such nucleic acid molecules can readily be identified as being able to hybridize under moderate to stringent conditions, to the nucleotide sequence shown in the Figure sheets or a fragment of the sequence.\",\n \"The gene encoding the novel Ras-like protein of the present invention is located on a genome component that has been mapped to human chromosome 6 (as indicated in FIG.\",\n \"3), which is supported by multiple lines of evidence, such as STS and BAC map data.\",\n \"FIG.\",\n \"3 provides information on SNPs that have been found in the gene encoding the Ras-like protein of the present invention.\",\n \"SNPs were identified at 37 different nucleotide positions.\",\n \"Some of these SNPs that are located outside the ORF and in introns may affect gene transcription.\",\n \"As used herein, the term “hybridizes under stringent conditions” is intended to describe conditions for hybridization and washing under which nucleotide sequences encoding a peptide at least 60-70% homologous to each other typically remain hybridized to each other.\",\n \"The conditions can be such that sequences at least about 60%, at least about 70%, or at least about 80% or more homologous to each other typically remain hybridized to each other.\",\n \"Such stringent conditions are known to those skilled in the art and can be found in Current Protocols in Molecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6.One example of stringent hybridization conditions are hybridization in 6× sodium chloridelsodium citrate (SSC) at about 45 C, followed by one or more washes in 0.2×SSC, 0.1% SDS at 50-65° C. Examples of moderate to low stringency hybridization conditions are well known in the art.\",\n \"Nucleic Acid Molecule Uses The nucleic acid molecules of the present invention are useful for probes, primers, chemical intermediates, and in biological assays.\",\n \"The nucleic acid molecules are useful as a hybridization probe for messenger RNA, transcript/cDNA and genomic DNA to isolate full-length cDNA and genomic clones encoding the peptide described in FIG.\",\n \"2 and to isolate cDNA and genomic clones that correspond to variants (alleles, orthologs, etc.)\",\n \"producing the same or related peptides shown in FIG.\",\n \"2.As illustrated in FIG.\",\n \"3, SNPs were identified at 37 different nucleotide positions.\",\n \"The probe can correspond to any sequence along the entire length of the nucleic acid molecules provided in the Figures.\",\n \"Accordingly, it could be derived from 5′ noncoding regions, the coding region, and 3′ noncoding regions.\",\n \"However, as discussed, fragments are not to be construed as those, which may encompass fragments disclosed prior to the present invention.\",\n \"The nucleic acid molecules are also useful as primers for PCR to amplify any given region of a nucleic acid molecule and are useful to synthesize antisense molecules of desired length and sequence.\",\n \"The nucleic acid molecules are also useful for constructing recombinant vectors.\",\n \"Such vectors include expression vectors that express a portion of, or all of, the peptide sequences.\",\n \"Vectors also include insertion vectors, used to integrate into another nucleic acid molecule sequence, such as into the cellular genome, to alter in situ expression of a gene and/or gene product.\",\n \"For example, an endogenous coding sequence can be replaced via homologous recombination with all or part of the coding region containing one or more specifically introduced mutations.\",\n \"The nucleic acid molecules are also useful for expressing antigenic portions of the proteins.\",\n \"The nucleic acid molecules are also useful as probes for determining the chromosomal positions of the nucleic acid molecules by means of in situ hybridization methods.\",\n \"The gene encoding the novel Ras-like protein of the present invention is located on a genome component that has been mapped to human chromosome 6 (as indicated in FIG.\",\n \"3), which is supported by multiple lines of evidence, such as STS and BAC map data.\",\n \"The nucleic acid molecules are also useful in making vectors containing the gene regulatory regions of the nucleic acid molecules of the present invention.\",\n \"The nucleic acid molecules are also useful for designing ribozymes corresponding to all, or a part, of the mRNA produced from the nucleic acid molecules described herein.\",\n \"The nucleic acid molecules are also useful for constructing host cells expressing a part, or all, of the nucleic acid molecules and peptides.\",\n \"Moreover, the nucleic acid molecules are useful for constructing transgenic animals wherein a homolog of the nucleic acid molecule has been “knocked-out” of the animal's genome.\",\n \"The nucleic acid molecules are also useful for constructing transgenic animals expressing all, or a part, of the nucleic acid molecules and peptides.\",\n \"The nucleic acid molecules are also useful for making vectors that express part, or all, of the peptides.\",\n \"The nucleic acid molecules are also useful as hybridization probes for determining the presence, level, form, and distribution of nucleic acid expression.\",\n \"Experimental data as provided in FIG.\",\n \"1 indicates that Ras-like proteins of the present invention are expressed in bone marrow and stem cells (as indicated by virtual northern blot analysis), and leukocytes (as indicated by PCR-based tissue screening panels).\",\n \"Accordingly, the probes can be used to detect the presence of, or to determine levels of, a specific nucleic acid molecule in cells, tissues, and in organisms.\",\n \"The nucleic acid whose level is determined can be DNA or RNA.\",\n \"Accordingly, probes corresponding to the peptides described herein can be used to assess expression and/or gene copy number in a given cell, tissue, or organism.\",\n \"These uses are relevant for diagnosis of disorders involving an increase or decrease in Ras-like protein expression relative to normal results.\",\n \"In vitro techniques for detection of mRNA include Northern hybridizations and in situ hybridizations.\",\n \"In vitro techniques for detecting DNA include Southern hybridizations and in situ hybridization.\",\n \"Probes can be used as a part of a diagnostic test kit for identifying cells or tissues that express a Ras-like protein, such as by measuring a level of a receptor-encoding nucleic acid in a sample of cells from a subject e.g., mRNA or genomic DNA, or determining if a receptor gene has been mutated.\",\n \"Experimental data as provided in FIG.\",\n \"1 indicates that Ras-like proteins of the present invention are expressed in bone marrow and stem cells (as indicated by virtual northern blot analysis), and leukocytes (as indicated by PCR-based tissue screening panels).\",\n \"Nucleic acid expression assays are useful for drug screening to identify compounds that modulate Ras-like protein nucleic acid expression.\",\n \"The invention thus provides a method for identifying a compound that can be used to treat a disorder associated with nucleic acid expression of the Ras-like protein gene, particularly biological and pathological processes that are mediated by the Ras-like protein in cells and tissues that express it.\",\n \"Experimental data as provided in FIG.\",\n \"1 indicates expression in bone marrow, stem cells, and leukocytes.\",\n \"The method typically includes assaying the ability of the compound to modulate the expression of the Ras-like protein nucleic acid and thus identifying a compound that can be used to treat a disorder characterized by undesired Ras-like protein nucleic acid expression.\",\n \"The assays can be performed in cell-based and cell-free systems.\",\n \"Cell-based assays include cells naturally expressing the Ras-like protein nucleic acid or recombinant cells genetically engineered to express specific nucleic acid sequences.\",\n \"The assay for Ras-like protein nucleic acid expression can involve direct assay of nucleic acid levels, such as mRNA levels, or on collateral compounds involved in the signal pathway.\",\n \"Further, the expression of genes that are up- or down-regulated in response to the Ras-like protein signal pathway can also be assayed.\",\n \"In this embodiment the regulatory regions of these genes can be operably linked to a reporter gene such as luciferase.\",\n \"Thus, modulators of Ras-like protein gene expression can be identified in a method wherein a cell is contacted with a candidate compound and the expression of mRNA determined.\",\n \"The level of expression of Ras-like protein mRNA in the presence of the candidate compound is compared to the level of expression of Ras-like protein mRNA in the absence of the candidate compound.\",\n \"The candidate compound can then be identified as a modulator of nucleic acid expression based on this comparison and be used, for example to treat a disorder characterized by aberrant nucleic acid expression.\",\n \"When expression of mRNA is statistically significantly greater in the presence of the candidate compound than in its absence, the candidate compound is identified as a stimulator of nucleic acid expression.\",\n \"When nucleic acid expression is statistically significantly less in the presence of the candidate compound than in its absence, the candidate compound is identified as an inhibitor of nucleic acid expression.\",\n \"The invention further provides methods of treatment, with the nucleic acid as a target, using a compound identified through drug screening as a gene modulator to modulate Ras-like protein nucleic acid expression in cells and tissues that express the Ras-like protein.\",\n \"Experimental data as provided in FIG.\",\n \"1 indicates that Ras-like proteins of the present invention are expressed in bone marrow and stem cells (as indicated by virtual northern blot analysis), and leukocytes (as indicated by PCR-based tissue screening panels).\",\n \"Modulation includes both up-regulation (i.e.\",\n \"activation or agonization) or down-regulation (suppression or antagonization) of nucleic acid expression.\",\n \"Alternatively, a modulator for Ras-like protein nucleic acid expression can be a small molecule or drug identified using the screening assays described herein as long as the drug or small molecule inhibits the Ras-like protein nucleic acid expression in the cells and tissues that express the protein.\",\n \"Experimental data as provided in FIG.\",\n \"1 indicates expression in bone marrow, stem cells, and leukocytes.\",\n \"The nucleic acid molecules are also useful for monitoring the effectiveness of modulating compounds on the expression or activity of the Ras-like protein gene in clinical trials or in a treatment regimen.\",\n \"Thus, the gene expression pattern can serve as a barometer for the continuing effectiveness of treatment with the compound, particularly with compounds to which a patient can develop resistance.\",\n \"The gene expression pattern can also serve as a marker indicative of a physiological response of the affected cells to the compound.\",\n \"Accordingly, such monitoring would allow either increased administration of the compound or the administration of alternative compounds to which the patient has not become resistant.\",\n \"Similarly, if the level of nucleic acid expression falls below a desirable level, administration of the compound could be commensurately decreased.\",\n \"The nucleic acid molecules are also useful in diagnostic assays for qualitative changes in Ras-like protein nucleic acid, and particularly in qualitative changes that lead to pathology.\",\n \"The nucleic acid molecules can be used to detect mutations in Ras-like protein genes and gene expression products such as mRNA.\",\n \"The nucleic acid molecules can be used as hybridization probes to detect naturally occurring genetic mutations in the Ras-like protein gene and thereby to determine whether a subject with the mutation is at risk for a disorder caused by the mutation.\",\n \"Mutations include deletion, addition, or substitution of one or more nucleotides in the gene, chromosomal rearrangement, such as inversion or transposition, modification of genomic DNA, such as aberrant methylation patterns, or changes in gene copy number, such as amplification.\",\n \"Detection of a mutated form of the Ras-like protein gene associated with a dysfunction provides a diagnostic tool for an active disease or susceptibility to disease when the disease results from overexpression, underexpression, or altered expression of a Ras-like protein.\",\n \"Individuals carrying mutations in the Ras-like protein gene can be detected at the nucleic acid level by a variety of techniques.\",\n \"FIG.\",\n \"3 provides information on SNPs that have been found in the gene encoding the Ras-like protein of the present invention.\",\n \"SNPs were identified at 37 different nucleotide positions.\",\n \"Some of these SNPs that are located outside the ORF and in introns may affect gene transcription.\",\n \"The gene encoding the novel Ras-like protein of the present invention is located on a genome component that has been mapped to human chromosome 6 (as indicated in FIG.\",\n \"3), which is supported by multiple lines of evidence, such as STS and BAC map data.\",\n \"Genomic DNA can be analyzed directly or can be amplified by using PCR prior to analysis.\",\n \"RNA or cDNA can be used in the same way.\",\n \"In some uses, detection of the mutation involves the use of a probe/primer in a polymerase chain reaction (PCR) (see, e.g.\",\n \"U.S. Pat.\",\n \"Nos.\",\n \"4,683,195 and 4,683,202), such as anchor PCR or RACE PCR, or, alternatively, in a ligation chain reaction (LCR) (see, e.g., Landegran et al., Science 241:1077-1080 (1988); and Nakazawa et al., PNAS 91:360-364 (1994)), the latter of which can be particularly useful for detecting point mutations in the gene (see Abravaya et al., Nucleic Acids Res.\",\n \"23:675-682 (1995)).\",\n \"This method can include the steps of collecting a sample of cells from a patient, isolating nucleic acid (e.g., genomic, mRNA or both) from the cells of the sample, contacting the nucleic acid sample with one or more primers which specifically hybridize to a gene under conditions such that hybridization and amplification of the gene (if present) occurs, and detecting the presence or absence of an amplification product, or detecting the size of the amplification product and comparing the length to a control sample.\",\n \"Deletions and insertions can be detected by a change in size of the amplified product compared to the normal genotype.\",\n \"Point mutations can be identified by hybridizing amplified DNA to normal RNA or antisense DNA sequences.\",\n \"Alternatively, mutations in a Ras-like protein gene can be directly identified, for example, by alterations in restriction enzyme digestion patterns determined by gel electrophoresis.\",\n \"Further, sequence-specific ribozymes (U.S. Pat.\",\n \"No.\",\n \"5,498,531) can be used to score for the presence of specific mutations by development or loss of a ribozyme cleavage site.\",\n \"Perfectly matched sequences can be distinguished from mismatched sequences by nuclease cleavage digestion assays or by differences in melting temperature.\",\n \"Sequence changes at specific locations can also be assessed by nuclease protection assays such as RNase and S1 protection or the chemical cleavage method.\",\n \"Furthermore, sequence differences between a mutant Ras-like protein gene and a wild-type gene can be determined by direct DNA sequencing.\",\n \"A variety of automated sequencing procedures can be utilized when performing the diagnostic assays (Naeve, C. W., Biotechniques 19:448 (1995)), including sequencing by mass spectrometry (see, e.g., PCT International Publication No.\",\n \"WO 94/16101; Cohen et al., Adv.\",\n \"Chromatogr.\",\n \"36:127-162 (1996); and Griffin et al., Appl.\",\n \"Biochem.\",\n \"Biotechnol.\",\n \"38:147-159 (1993)).\",\n \"Other methods for detecting mutations in the gene include methods in which protection from cleavage agents is used to detect mismatched bases in RNA/RNA or RNA/DNA duplexes (Myers et al., Science 230:1242 (1985)); Cotton et al., PNAS 85:4397 (1988); Saleeba et al., Meth.\",\n \"Enzymol.\",\n \"217:286-295 (1992)), electrophoretic mobility of mutant and wild type nucleic acid is compared (Orita et al., PNAS86:2766 (1989); Cotton et al., Mutat.\",\n \"Res.\",\n \"285:125-144 (1993); and Hayashi et al., Genet.\",\n \"Anal.\",\n \"Tech Appl.\",\n \"9:73-79 (1992)), and movement of mutant or wild-type fragments in polyacrylamide gels containing a gradient of denaturant is assayed using denaturing gradient gel electrophoresis (Myers et al., Nature 313:495 (1985)).\",\n \"Examples of other techniques for detecting point mutations include, selective oligonucleotide hybridization, selective amplification, and selective primer extension.\",\n \"The nucleic acid molecules are also useful for testing an individual for a genotype that while not necessarily causing the disease, nevertheless affects the treatment modality.\",\n \"Thus, the nucleic acid molecules can be used to study the relationship between an individual's genotype and the individual's response to a compound used for treatment (pharmacogenomic relationship).\",\n \"Accordingly, the nucleic acid molecules described herein can be used to assess the mutation content of the Ras-like protein gene in an individual in order to select an appropriate compound or dosage regimen for treatment.\",\n \"FIG.\",\n \"3 provides information on SNPs that have been found in the gene encoding the Ras-like protein of the present invention.\",\n \"SNPs were identified at 37 different nucleotide positions.\",\n \"Some of these SNPs that are located outside the ORF and in introns may affect gene transcription.\",\n \"Thus nucleic acid molecules displaying genetic variations that affect treatment provide a diagnostic target that can be used to tailor treatment in an individual.\",\n \"Accordingly, the production of recombinant cells and animals containing these polymorphisms allow effective clinical design of treatment compounds and dosage regimens.\",\n \"The nucleic acid molecules are thus useful as antisense constructs to control Ras-like protein gene expression in cells, tissues, and organisms.\",\n \"A DNA antisense nucleic acid molecule is designed to be complementary to a region of the gene involved in transcription, preventing transcription and hence production of Ras-like protein.\",\n \"An antisense RNA or DNA nucleic acid molecule would hybridize to the mRNA and thus block translation of mRNA into Ras-like protein.\",\n \"Alternatively, a class of antisense molecules can be used to inactivate mRNA in order to decrease expression of Ras-like protein nucleic acid.\",\n \"Accordingly, these molecules can treat a disorder characterized by abnormal or undesired Ras-like protein nucleic acid expression.\",\n \"This technique involves cleavage by means of ribozymes containing nucleotide sequences complementary to one or more regions in the mRNA that attenuate the ability of the mRNA to be translated.\",\n \"Possible regions include coding regions and particularly coding regions corresponding to the catalytic and other functional activities of the Ras-like protein, such as ligand binding.\",\n \"The nucleic acid molecules also provide vectors for gene therapy in patients containing cells that are aberrant in Ras-like protein gene expression.\",\n \"Thus, recombinant cells, which include the patient's cells that have been engineered ex vivo and returned to the patient, are introduced into an individual where the cells produce the desired Ras-like protein to treat the individual.\",\n \"The invention also encompasses kits for detecting the presence of a Ras-like protein nucleic acid in a biological sample.\",\n \"Experimental data as provided in FIG.\",\n \"1 indicates that Ras-like proteins of the present invention are expressed in bone marrow and stem cells (as indicated by virtual northern blot analysis), and leukocytes (as indicated by PCR-based tissue screening panels).\",\n \"For example, the kit can comprise reagents such as a labeled or labelable nucleic acid or agent capable of detecting Ras-like protein nucleic acid in a biological sample; means for determining the amount of Ras-like protein nucleic acid in the sample; and means for comparing the amount of Ras-like protein nucleic acid in the sample with a standard.\",\n \"The compound or agent can be packaged in a suitable container.\",\n \"The kit can further comprise instructions for using the kit to detect Ras-like protein mRNA or DNA.\",\n \"Nucleic Acid Arrays The present invention further provides arrays or microarrays of nucleic acid molecules that are based on the sequence information provided in FIGS.\",\n \"1 and 3 (SEQ ID NOS:1 and 3).\",\n \"As used herein “Arrays” or “Microarrays” refers to an array of distinct polynucleotides or oligonucleotides synthesized on a substrate, such as paper, nylon or other type of membrane, filter, chip, glass slide, or any other suitable solid support.\",\n \"In one embodiment, the microarray is prepared and used according to the methods described in U.S. Pat.\",\n \"No.\",\n \"5,837,832, Chee et al., PCT application WO95/11995 (Chee et al.\",\n \"), Lockhart, D. J. et al.\",\n \"(1996; Nat.\",\n \"Biotech.\",\n \"14: 1675-1680) and Schena, M. et al.\",\n \"(1996; Proc.\",\n \"Natl.\",\n \"Acad.\",\n \"Sci.\",\n \"93: 10614-10619), all of which are incorporated herein in their entirety by reference.\",\n \"In other embodiments, such arrays are produced by the methods described by Brown et.\",\n \"al., U.S. Pat.\",\n \"No.\",\n \"5,807,522.The microarray is preferably composed of a large number of unique, single-stranded nucleic acid sequences, usually either synthetic antisense oligonucleotides or fragments of cDNAs, fixed to a solid support.\",\n \"The oligonucleotides are preferably about 6-60 nucleotides in length, more preferably 15-30 nucleotides in length, and most preferably about 20-25 nucleotides in length.\",\n \"For a certain type of microarray, it may be preferable to use oligonucleotides that are only 7-20 nucleotides in length.\",\n \"The microarray may contain oligonucleotides that cover the known 5′, or 3′, sequence, sequential oligonucleotides that cover the full-length sequence; or unique oligonucleotides selected from particular areas along the length of the sequence.\",\n \"Polynucleotides used in the microarray may be oligonucleotides that are specific to a gene or genes of interest.\",\n \"In order to produce oligonucleotides to a known sequence for a microarray, the gene(s) of interest (or an ORF identified from the contigs of the present invention) is typically examined using a computer algorithm that starts at the 5′ or at the 3′ end of the nucleotide sequence.\",\n \"Typical algorithms will then identify oligomers of defined length that are unique to the gene, have a GC content within a range suitable for hybridization, and lack predicted secondary structure that may interfere with hybridization.\",\n \"In certain situations it may be appropriate to use pairs of oligonucleotides on a microarray.\",\n \"The “pairs” will be identical, except for one nucleotide that preferably is located in the center of the sequence.\",\n \"The second oligonucleotide in the pair (mismatched by one) serves as a control.\",\n \"The number of oligonucleotide pairs may range from two to one million.\",\n \"The oligomers are synthesized at designated areas on a substrate using a light-directed chemical process.\",\n \"The substrate may be paper, nylon or other type of membrane, filter, chip, glass slide or any other suitable solid support.\",\n \"In another aspect, an oligonucleotide may be synthesized on the surface of the substrate by using a chemical coupling procedure and an ink jet application apparatus, as described in PCT application WO95/251116 (Baldeschweiler et al.)\",\n \"which is incorporated herein in its entirety by reference.\",\n \"In another aspect, a “gridded” array analogous to a dot (or slot) blot may be used to arrange and link cDNA fragments or oligonucleotides to the surface of a substrate using a vacuum system, thermal, UV, mechanical or chemical bonding procedures.\",\n \"An array, such as those described above, may be produced by hand or by using available devices (slot blot or dot blot apparatus), materials (any suitable solid support), and machines (including robotic instruments), and may contain 8, 24, 96, 384, 1536, 6144 or more oligonucleotides, or any other number between two and one million which lends itself to the efficient use of commercially available instrumentation.\",\n \"In order to conduct sample analysis using a microarray, the RNA or DNA from a biological sample is made into hybridization probes.\",\n \"The mRNA is isolated, and cDNA is produced and used as a template to make antisense RNA (aRNA).\",\n \"The aRNA is amplified in the presence of fluorescent nucleotides, and labeled probes are incubated with the microarray so that the probe sequences hybridize to complementary oligonucleotides of the microarray.\",\n \"Incubation conditions are adjusted so that hybridization occurs with precise complementary matches or with various degrees of less complementarity.\",\n \"After removal of nonhybridized probes, a scanner is used to determine the levels and patterns of fluorescence.\",\n \"The scanned images are examined to determine degree of complementarity and the relative abundance of each oligonucleotide sequence on the microarray.\",\n \"The biological samples may be obtained from any bodily fluids (such as blood, urine, saliva, phlegm, gastric juices, etc.\",\n \"), cultured cells, biopsies, or other tissue preparations.\",\n \"A detection system may be used to measure the absence, presence, and amount of hybridization for all of the distinct sequences simultaneously.\",\n \"This data may be used for large-scale correlation studies on the sequences, expression patterns, mutations, variants, or polymorphisms among samples.\",\n \"Using such arrays, the present invention provides methods to identify the expression of one or more of the proteins/peptides of the present invention.\",\n \"In detail, such methods comprise incubating a test sample with one or more nucleic acid molecules and assaying for binding of the nucleic acid molecule with components within the test sample.\",\n \"Such assays will typically involve arrays comprising many genes, at least one of which is a gene of the present invention.\",\n \"FIG.\",\n \"3 provides information on SNPs that have been found in the gene encoding the Ras-like protein of the present invention.\",\n \"SNPs were identified at 37 different nucleotide positions.\",\n \"Some of these SNPs that are located outside the ORF and in introns may affect gene transcription.\",\n \"Conditions for incubating a nucleic acid molecule with a test sample vary.\",\n \"Incubation conditions depend on the format employed in the assay, the detection methods employed, and the type and nature of the nucleic acid molecule used in the assay.\",\n \"One skilled in the art will recognize that any one of the commonly available hybridization, amplification or array assay formats can readily be adapted to employ the novel fragments of the human genome disclosed herein.\",\n \"Examples of such assays can be found in Chard, T, An Introduction to Radioimmunoassay and Related Techniques, Elsevier Science Publishers, Amsterdam, The Netherlands (1986); Bullock, G. R. et al., Techniques in Immunocytochemistry, Academic Press, Orlando, Fla. Vol.\",\n \"1 (1982), Vol.\",\n \"2 (1983), Vol.\",\n \"3 (1985); Tijssen, P., Practice and Theory of Enzyme Immunoassays: Laboratory Techniques in Biochemistry and Molecular Biology, Elsevier Science Publishers, Amsterdam, The Netherlands (1985).\",\n \"The test samples of the present invention include cells, protein or membrane extracts of cells.\",\n \"The test sample used in the above-described method will vary based on the assay format, nature of the detection method and the tissues, cells or extracts used as the sample to be assayed.\",\n \"Methods for preparing nucleic acid extracts or of cells are well known in the art and can be readily be adapted in order to obtain a sample that is compatible with the system utilized.\",\n \"In another embodiment of the present invention, kits are provided which contain the necessary reagents to carry out the assays of the present invention.\",\n \"Specifically, the invention provides a compartmentalized kit to receive, in close confinement, one or more containers which comprises: (a) a first container comprising one of the nucleic acid molecules that can bind to a fragment of the human genome disclosed herein; and (b) one or more other containers comprising one or more of the following: wash reagents, reagents capable of detecting presence of a bound nucleic acid.\",\n \"Preferred kits will include chips that are capable of detecting the expression of 10 or more, 100 or more, or 500 or more, 1000 or more, or all of the genes expressed in Human.\",\n \"In detail, a compartmentalized kit includes any kit in which reagents are contained in separate containers.\",\n \"Such containers include small glass containers, plastic containers, strips of plastic, glass or paper, or arraying material such as silica.\",\n \"Such containers allows one to efficiently transfer reagents from one compartment to another compartment such that the samples and reagents are not cross-contaminated, and the agents or solutions of each container can be added in a quantitative fashion from one compartment to another.\",\n \"Such containers will include a container which will accept the test sample, a container which contains the nucleic acid probe, containers which contain wash reagents (such as phosphate buffered saline, Tris-buffers, etc.\",\n \"), and containers which contain the reagents used to detect the bound probe.\",\n \"One skilled in the art will readily recognize that the previously unidentified Ras-like protein genes of the present invention can be routinely identified using the sequence information disclosed herein can be readily incorporated into one of the established kit formats which are well known in the art, particularly expression arrays.\",\n \"Vectors/Host Cells The invention also provides vectors containing the nucleic acid molecules described herein.\",\n \"The term “vector” refers to a vehicle, preferably a nucleic acid molecule, which can transport the nucleic acid molecules.\",\n \"When the vector is a nucleic acid molecule, the nucleic acid molecules are covalently linked to the vector nucleic acid.\",\n \"With this aspect of the invention, the vector includes a plasmid, single or double stranded phage, a single or double stranded RNA or DNA viral vector, or artificial chromosome, such as a BAC, PAC, YAC, OR MAC.\",\n \"A vector can be maintained in the host cell as an extrachromosomal element where it replicates and produces additional copies of the nucleic acid molecules.\",\n \"Alternatively, the vector may integrate into the host cell genome and produce additional copies of the nucleic acid molecules when the host cell replicates.\",\n \"The invention provides vectors for the maintenance (cloning vectors) or vectors for expression (expression vectors) of the nucleic acid molecules.\",\n \"The vectors can function in procaryotic or eukaryotic cells or in both (shuttle vectors).\",\n \"Expression vectors contain cis-acting regulatory regions that are operably linked in the vector to the nucleic acid molecules such that transcription of the nucleic acid molecules is allowed in a host cell.\",\n \"The nucleic acid molecules can be introduced into the host cell with a separate nucleic acid molecule capable of affecting transcription.\",\n \"Thus, the second nucleic acid molecule may provide a trans-acting factor interacting with the cis-regulatory control region to allow transcription of the nucleic acid molecules from the vector.\",\n \"Alternatively, a trans-acting factor may be supplied by the host cell.\",\n \"Finally, a trans-acting factor can be produced from the vector itself.\",\n \"It is understood, however, that in some embodiments, transcription and/or translation of the nucleic acid molecules can occur in a cell-free system.\",\n \"The regulatory sequence to which the nucleic acid molecules described herein can be operably linked include promoters for directing mRNA transcription.\",\n \"These include, but are not limited to, the left promoter from bacteriophage λ, the lac, TRP, and TAC promoters from E. coli, the early and late promoters from SV40, the CMV immediate early promoter, the adenovirus early and late promoters, and retrovirus long-terminal repeats.\",\n \"In addition to control regions that promote transcription, expression vectors may also include regions that modulate transcription, such as repressor binding sites and enhancers.\",\n \"Examples include the SV40 enhancer, the cytomegalovirus immediate early enhancer, polyoma enhancer, adenovirus enhancers, and retrovirus LTR enhancers.\",\n \"In addition to containing sites for transcription initiation and control, expression vectors can also contain sequences necessary for transcription termination and, in the transcribed region a ribosome binding site for translation.\",\n \"Other regulatory control elements for expression include initiation and termination codons as well as polyadenylation signals.\",\n \"The person of ordinary skill in the art would be aware of the numerous regulatory sequences that are useful in expression vectors.\",\n \"Such regulatory sequences are described, for example, in Sambrook et al., Molecular Cloning: A Laboratory Manual.\",\n \"2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., (1989).\",\n \"A variety of expression vectors can be used to express a nucleic acid molecule.\",\n \"Such vectors include chromosomal, episomal, and virus-derived vectors, for example vectors derived from bacterial plasmids, from bacteriophage, from yeast episomes, from yeast chromosomal elements, including yeast artificial chromosomes, from viruses such as baculoviruses, papovaviruses such as SV40, Vaccinia viruses, adenoviruses, poxviruses, pseudorabies viruses, and retroviruses.\",\n \"Vectors may also be derived from combinations of these sources such as those derived from plasmid and bacteriophage genetic elements, e.g.\",\n \"cosmids and phagemids.\",\n \"Appropriate cloning and expression vectors for prokaryotic and eukaryotic hosts are described in Sambrook et al., Molecular Cloning: A Laboratory Manual.\",\n \"2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., (1989).\",\n \"The regulatory sequence may provide constitutive expression in one or more host cells (i.e.\",\n \"tissue specific) or may provide for inducible expression in one or more cell types such as by temperature, nutrient additive, or exogenous factor such as a hormone or other ligand.\",\n \"A variety of vectors providing for constitutive and inducible expression in prokaryotic and eukaryotic hosts are well known to those of ordinary skill in the art.\",\n \"The nucleic acid molecules can be inserted into the vector nucleic acid by well-known methodology.\",\n \"Generally, the DNA sequence that will ultimately be expressed is joined to an expression vector by cleaving the DNA sequence and the expression vector with one or more restriction enzymes and then ligating the fragments together.\",\n \"Procedures for restriction enzyme digestion and ligation are well known to those of ordinary skill in the art.\",\n \"The vector containing the appropriate nucleic acid molecule can be introduced into an appropriate host cell for propagation or expression using well-known techniques.\",\n \"Bacterial cells include, but are not limited to, E. coli, Streptomyces, and Salmonella typhimurium.\",\n \"Eukaryotic cells include, but are not limited to, yeast, insect cells such as Drosophila, animal cells such as COS and CHO cells, and plant cells.\",\n \"As described herein, it may be desirable to express the peptide as a fusion protein.\",\n \"Accordingly, the invention provides fusion vectors that allow for the production of the peptides.\",\n \"Fusion vectors can increase the expression of a recombinant protein, increase the solubility of the recombinant protein, and aid in the purification of the protein by acting for example as a ligand for affinity purification.\",\n \"A proteolytic cleavage site may be introduced at the junction of the fusion moiety so that the desired peptide can ultimately be separated from the fusion moiety.\",\n \"Proteolytic enzymes include, but are not limited to, factor Xa, thrombin, and enteroRas-like protein.\",\n \"Typical fusion expression vectors include pGEX (Smith et al., Gene 67:3140 (1988)), pMAL (New England Biolabs, Beverly, Mass.)\",\n \"and pRIT5 (Pharmacia, Piscataway, N.J.) which fuse glutathione S-transferase (GST), maltose E binding protein, or protein A, respectively, to the target recombinant protein.\",\n \"Examples of suitable inducible non-fusion E. coli expression vectors include pTrc (Amann et al., Gene 69:301-315 (1988)>and pET 11d (Studier et al., Gene Expression Technology: Methods in Enzymology 185:60-89 (1990)).\",\n \"Recombinant protein expression can be maximized in a host bacteria by providing a genetic background wherein the host cell has an impaired capacity to proteolytically cleave the recombinant protein.\",\n \"(Gottesman, S., Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, Calif. (1990)119-128).\",\n \"Alternatively, the sequence of the nucleic acid molecule of interest can be altered to provide preferential codon usage for a specific host cell, for example E. coli.\",\n \"(Wada et al., Nucleic Acids Res.\",\n \"20:2111-2118 (1992)).\",\n \"The nucleic acid molecules can also be expressed by expression vectors that are operative in yeast.\",\n \"Examples of vectors for expression in yeast e.g., S. cerevisiae include pYepSec1 (Baldari, et al., EMBO J.\",\n \"6:229-234 (1987)), pMFa (Kurjan et al., Cell 30:933-943(1982)), pJRY88 (Schultz et al., Gene 54:113-123 (1987)), and pYES2 (Invitrogen Corporation, San Diego, Calif.).\",\n \"The nucleic acid molecules can also be expressed in insect cells using, for example, baculovirus expression vectors.\",\n \"Baculovirus vectors available for expression of proteins in cultured insect cells (e.g., Sf 9 cells) include the pAc series (Smith et al., Mol.\",\n \"Cell Biol.\",\n \"3:2156-2165 (1983)) and the pVL series (Lucklow et al., Virology 170:31-39 (1989)).\",\n \"In certain embodiments of the invention, the nucleic acid molecules described herein are expressed in mammalian cells using mammalian expression vectors.\",\n \"Examples of mammalian expression vectors include pCDM8 (Seed, B.\",\n \"Nature 329:840(1987)) and pMT2PC (Kaufman et al., EMBO J.\",\n \"6:187-195 (1987)).\",\n \"The expression vectors listed herein are provided by way of example only of the well-known vectors available to those of ordinary skill in the art that would be useful to express the nucleic acid molecules.\",\n \"The person of ordinary skill in the art would be aware of other vectors suitable for maintenance, propagation, or expression of the nucleic acid molecules described herein.\",\n \"These are found for example in Sambrook, J., Fritsh, E. F., and Maniatis, T. Molecular Cloning: A Laboratory Manual.\",\n \"2nd, ed, Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989.The invention also encompasses vectors in which the nucleic acid sequences described herein are cloned into the vector in reverse orientation, but operably linked to a regulatory sequence that permits transcription of antisense RNA.\",\n \"Thus, an antisense transcript can be produced to all, or to a portion, of the nucleic acid molecule sequences described herein, including both coding and non-coding regions.\",\n \"Expression of this antisense RNA is subject to each of the parameters described above in relation to expression of the sense RNA (regulatory sequences, constitutive or inducible expression, tissue-specific expression).\",\n \"The invention also relates to recombinant host cells containing the vectors described herein.\",\n \"Host cells therefore include prokaryotic cells, lower eukaryotic cells such as yeast, other eukaryotic cells such as insect cells, and higher eukaryotic cells such as mammalian cells.\",\n \"The recombinant host cells are prepared by introducing the vector constructs described herein into the cells by techniques readily available to the person of ordinary skill in the art.\",\n \"These include, but are not limited to, calcium phosphate transfection, DEAE-dextran-mediated transfection, cationic lipid-mediated transfection, electroporation, transduction, infection, lipofection, and other techniques such as those found in Sambrook, et al.\",\n \"(Molecular Cloning: A Laboratory Manual.\",\n \"2nd, ed, Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989).\",\n \"Host cells can contain more than one vector.\",\n \"Thus, different nucleotide sequences can be introduced on different vectors of the same cell.\",\n \"Similarly, the nucleic acid molecules can be introduced either alone or with other nucleic acid molecules that are not related to the nucleic acid molecules such as those providing trans-acting factors for expression vectors.\",\n \"When more than one vector is introduced into a cell, the vectors can be introduced independently, co-introduced, or joined to the nucleic acid molecule vector.\",\n \"In the case of bacteriophage and viral vectors, these can be introduced into cells as packaged or encapsulated virus by standard procedures for infection and transduction.\",\n \"Viral vectors can be replication-competent or replication-defective.\",\n \"In the case in which viral replication is defective, replication will occur in host cells providing functions that complement the defects.\",\n \"Vectors generally include selectable markers that enable the selection of the subpopulation of cells that contain the recombinant vector constructs.\",\n \"The marker can be contained in the same vector that contains the nucleic acid molecules described herein or may be on a separate vector.\",\n \"Markers include tetracycline or ampicillin-resistance genes for prokaryotic host cells and dihydrofolate reductase or neomycin resistance for eukaryotic host cells.\",\n \"However, any marker that provides selection for a phenotypic trait will be effective.\",\n \"While the mature proteins can be produced in bacteria, yeast, mammalian cells, and other cells under the control of the appropriate regulatory sequences, cell-free transcription and translation systems can also be used to produce these proteins using RNA derived from the DNA constructs described herein.\",\n \"Where secretion of the peptide is desired, which is difficult to achieve with multi-transmembrane domain containing proteins such as kinases, appropriate secretion signals are incorporated into the vector.\",\n \"The signal sequence can be endogenous to the peptides or heterologous to these peptides.\",\n \"Where the peptide is not secreted into the medium, which is typically the case with kinases, the protein can be isolated from the host cell by standard disruption procedures, including freeze thaw, sonication, mechanical disruption, use of lysing agents and the like.\",\n \"The peptide can then be recovered and purified by well-known purification methods including ammonium sulfate precipitation, acid extraction, anion or cationic exchange chromatography, phosphocellulose chromatography, hydrophobic-interaction chromatography, affinity chromatography, hydroxylapatite chromatography, lectin chromatography, or high performance liquid chromatography.\",\n \"It is also understood that depending upon the host cell in recombinant production of the peptides described herein, the peptides can have various glycosylation patterns, depending upon the cell, or maybe non-glycosylated as when produced in bacteria.\",\n \"In addition, the peptides may include an initial modified methionine in some cases as a result of a host-mediated process.\",\n \"Uses of Vectors and Host Cells The recombinant host cells expressing the peptides described herein have a variety of uses.\",\n \"First, the cells are useful for producing a Ras-like protein polypeptide that can be further purified to produce desired amounts of Ras-like protein or fragments.\",\n \"Thus, host cells containing expression vectors are useful for peptide production.\",\n \"Host cells are also useful for conducting cell-based assays involving the Ras-like protein or Ras-like protein fragments.\",\n \"Thus, a recombinant host cell expressing a native Ras-like protein is useful for assaying compounds that stimulate or inhibit Ras-like protein function.\",\n \"Host cells are also useful for identifying Ras-like protein mutants in which these functions are affected.\",\n \"If the mutants naturally occur and give rise to a pathology, host cells containing the mutations are useful to assay compounds that have a desired effect on the mutant Ras-like protein (for example, stimulating or inhibiting function) which may not be indicated by their effect on the native Ras-like protein.\",\n \"Genetically engineered host cells can be further used to produce non-human transgenic animals.\",\n \"A transgenic animal is preferably a mammal, for example a rodent, such as a rat or mouse, in which one or more of the cells of the animal include a transgene.\",\n \"A transgene is exogenous DNA which is integrated into the genome of a cell from which a transgenic animal develops and which remains in the genome of the mature animal in one or more cell types or tissues of the transgenic animal.\",\n \"These animals are useful for studying the function of a Ras-like protein and identifying and evaluating modulators of Ras-like protein activity.\",\n \"Other examples of transgenic animals include non-human primates, sheep, dogs, cows, goats, chickens, and amphibians.\",\n \"A transgenic animal can be produced by introducing nucleic acid into the male pronuclei of a fertilized oocyte, e.g., by microinjection, retroviral infection, and allowing the oocyte to develop in a pseudopregnant female foster animal.\",\n \"Any of the Ras-like protein nucleotide sequences can be introduced as a transgene into the genome of a non-human animal, such as a mouse.\",\n \"Any of the regulatory or other sequences useful in expression vectors can form part of the transgenic sequence.\",\n \"This includes intronic sequences and polyadenylation signals, if not already included.\",\n \"A tissue-specific regulatory sequence(s) can be operably linked to the transgene to direct expression of the Ras-like protein to particular cells.\",\n \"Methods for generating transgenic animals via embryo manipulation and microinjection, particularly animals such as mice, have become conventional in the art and are described, for example, in U.S. Pat.\",\n \"Nos.\",\n \"4,736,866 and 4,870,009, both by Leder et al., U.S. Pat.\",\n \"No.\",\n \"4,873,191 by Wagner et al.\",\n \"and in Hogan, B., Manipulating the Mouse Embryo, (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1986).\",\n \"Similar methods are used for production of other transgenic animals.\",\n \"A transgenic founder animal can be identified based upon the presence of the transgene in its genome and/or expression of transgenic mRNA in tissues or cells of the animals.\",\n \"A transgenic founder animal can then be used to breed additional animals carrying the transgene.\",\n \"Moreover, transgenic animals carrying a transgene can further be bred to other transgenic animals carrying other transgenes.\",\n \"A transgenic animal also includes animals in which the entire animal or tissues in the animal have been produced using the homologously recombinant host cells described herein.\",\n \"In another embodiment, transgenic non-human animals can be produced which contain selected systems which allow for regulated expression of the transgene.\",\n \"One example of such a system is the cre/loxP recombinase system of bacteriophage P1.For a description of the cre/loxP recombinase system, see, e.g., Lakso et al.\",\n \"PNAS 89:6232-6236 (1992).\",\n \"Another example of a recombinase system is the FLP recombinase system of S. cerevisiae (O'Gorman et al.\",\n \"Science 251:1351-1355 (1991).\",\n \"If a cre/loxP recombinase system is used to regulate expression of the transgene, animals containing transgenes encoding both the Cre recombinase and a selected protein is required.\",\n \"Such animals can be provided through the construction of “double” transgenic animals, e.g., by mating two transgenic animals, one containing a transgene encoding a selected protein and the other containing a transgene encoding a recombinase.\",\n \"Clones of the non-human transgenic animals described herein can also be produced according to the methods described in Wilmut, I. et al.\",\n \"Nature 385:810-813 (1997) and PCT International Publication Nos.\",\n \"WO 97/07668 and WO 97/07669.In brief, a cell, e.g., a somatic cell, from the transgenic animal can be isolated and induced to exit the growth cycle and enter Go phase.\",\n \"The quiescent cell can then be fused, e.g., through the use of electrical pulses, to an enucleated oocyte from an animal of the same species from which the quiescent cell is isolated.\",\n \"The reconstructed oocyte is then cultured such that it develops to morula or blastocyst and then transferred to pseudopregnant female foster animal.\",\n \"The offspring born of this female foster animal will be a clone of the animal from which the cell, e.g., the somatic cell, is isolated.\",\n \"Transgenic animals containing recombinant cells that express the peptides described herein are useful to conduct the assays described herein in an in vivo context.\",\n \"Accordingly, the various physiological factors that are present in vivo and that could effect ligand binding, Ras-like protein activation, and signal transduction, may not be evident from in vitro cell-free or cell-based assays.\",\n \"Accordingly, it is useful to provide non-human transgenic animals to assay in vivo Ras-like protein function, including ligand interaction, the effect of specific mutant Ras-like proteins on Ras-like protein function and ligand interaction, and the effect of chimeric Ras-like proteins.\",\n \"It is also possible to assess the effect of null mutations, which is mutations that substantially or completely eliminate one or more Ras-like protein functions.\",\n \"All publications and patents mentioned in the above specification are herein incorporated by reference.\",\n \"Various modifications and variations of the described method and system of the invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention.\",\n \"Although the invention has been described in connection with specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments.\",\n \"Indeed, various modifications of the above-described modes for carrying out the invention, which are obvious to those skilled in the field of molecular biology or related fields, are intended to be within the scope of the following claims.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473737"}}},{"rowIdx":747,"cells":{"text":{"kind":"list like","value":[["Novel modulator of non-genomic activity of nuclear receptors (mnar) and uses thereof","A novel protein modulator of non-genomic activity of nuclear receptors (MNAR), which interacts with estrogen receptors alpha (ERα), estrogen receptor beta (ERβ), and other nuclear receptors, was discovered and characterized.","An MNAR-ER complex, in vitro and in vivo, relates to nucleic acids, antisense, recombinant expression of nucleic acids, as well as host cells, compositions and assays that utilize the nucleic acids and protein of this invention."],["1.An isolated nucleic acid sequence comprising a polynucleotide selected from the group consisting of: (a) a polynucleotide encoding a novel protein modulator of non-genomic activity of nuclear receptors (MNAR) comprising the amino acid sequence of SEQ ID NO:2 or of SEQ ID NO:13; (b) a polynucleotide that hybridizes under highly stringent conditions with (i) a region of the nucleotide sequence of SEQ ID NO: 1 or of SEQ ID NO:12, (ii) a subsequence of at least 100 nucleotides of the nucleotide sequence of SEQ ID NO: 1 or of SEQ ID NO:12, (iii) or a complementary strand of (i) or (ii); (c) a polynucleotide comprising a sequence with at least 85% identity to a polynucleotide coding sequence of SEQ ID NO: 1 or of SEQ ID NO: 12; (d) a variant of the polynucleotide comprising a polynucleotide coding sequence of SEQ ID NO:1 or of SEQ ID NO:12; and (e) a polynucleotide encoding a polypeptide fragment comprising the amino acids of SEQ ID NOS: 20 or 23 (f) a complement or antisense form of the polynucleotide of (a), (b), (c), (d) or (e); wherein the polynucleotide does not encode a p160 protein: 2.The isolated nucleic acid sequence of claim 1, wherein the polynucleotide sequence is an allelic variant.","3.A polypeptide comprising an amino acid sequence selected from the group consisting of: (a) an amino acid sequence for a protein modulator of non-genomic activity of nuclear receptors (MNAR) comprising the amino acid sequence of SEQ ID NO: 2 or of SEQ ID NO:13; (b) an amino acid sequence for an MNAR fragment comprising the amino acid sequence of SEQ ID NOS:14, 15 or 16; (c) a variant of (a) or (b), and (d) a fragment of (a), (b), or (c).","4.The polypeptide of claim 3 comprising one or more amino acid sequences which are more than 90% identical to the amino acid sequence of SEQ ID NO:14.5.The polypeptide of claim 3 comprising one or more amino acid sequences which are more than 95% identical to the mutant sequence of SEQ ID NO:14.6.The polypeptide of claim 3, wherein said polypeptide comprises the amino acid sequence of SEQ ID NOS 2, 13, 14, 15 16 20 or 22.7.A fusion polypeptide comprising a polypeptide, or polypeptide fragment of MNAR.","8.The polypeptide according to claim 3, wherein said polypeptide stimulates the activity of a tyrosine kinase in the presence of one or more nuclear receptors.","9.The polypeptide of claim 8, wherein said nuclear receptor is a steroid receptor.","10.The polypeptide of claim 8, wherein said nuclear receptor is an estrogen receptor.","11.The polypeptide of claim 8, wherein said estrogen receptor is estrogen receptor alpha.","12.An expression vector comprising the isolated nucleic acid sequence of claim 1.13.The expression vector of claim 12 comprising the polynucleotide sequence of SED ID NO: 1 or of SEQ ID NO:12.14.A host cell transformed with the vector of claim 12.15.The host cell of claim 14, wherein the host cell is a mammalian host cell.","16.An isolated antibody to the polypeptide of claim 3.17.The antibody of claim 16, wherein said antibody is a monoclonal antibody.","18.The antibody of claim 16, wherein said antibody is a polyclonal antibody.","19.A method for modulating the transcriptional activity of a nuclear receptor, comprising providing to the loci of a nuclear receptor a polypeptide according to claim 3.20.A method for modulating the transcriptional activity of a nuclear receptor, comprising: (a) transforming a host cell with the recombinant vector having the nucleic acid sequence for MNAR according to claim 1; and (b) culturing said transformed host cell to express the nucleic acid in the presence of a nuclear receptor.","21.A method of identifying compounds exhibiting non-genomic activity versus genomic activity comprising administering a test compound to a cell comprising an MNAR-nuclear receptor complex and measuring the non-genomic or genomic activity resulting from the test compound.","22.The method of claim 21, wherein at least one ligand of ER is present in the cell, or a kinase is present, or both are present.","23.A method of screening for a compound that modulates the activity of MNAR on a nuclear receptor, comprising the steps of (a) contacting a test compound with the polypeptide of claim 3; and (b) determining whether said test compound specifically binds said polypeptide.","24.A method of screening for a compound that modulates the activity of MNAR on a nuclear receptor, said method comprising the steps of (a) adding a test compound to a cell comprising the polypeptide of claim 3 and a nuclear receptor; and (b) comparing the MNAR activity before and after said adding step.","25.The method of claim 24, said method comprising adding a test compound to a control comprising a mutant cell lacking MNAR activity or with significantly reduced MNAR activity.","26.A method for detecting the non-genomic activity of a compound comprising administering a compound to a cell in the presence of MNAR-nuclear receptor and in the absence of MNAR and comparing the level of non-genomic activity.","27.A method for detecting the genomic activity of a compound comprising administering a compound to a cell comprising a MNAR-nuclear receptor complex and comparing the level of genomic activity in the absence of MNAR.","28.A method for detecting the genomic activity of a compound according to claim 51 wherein the MNAR-ER cell overexpresses MNAR.","29.A method for detecting the genomic activity of a compound according to claim 51 wherein the MNAR-ER cell overexpresses ER.","30.A method for detecting the genomic activity of a compound according to claim 27 wherein the ER is operatively associated with a reporter.","31.A method of identifying compounds having selective genomic versus non-genomic activity said method comprising the steps of (a) adding a test compound to a cell comprising a MNAR-nuclear receptor complex; and (b) comparing the genomic versus non-genomic activity before and after said adding the test compound.","32.The method of claim 59, wherein selective genomic activity is measured as a two-fold increase after addition of said test compound to a cell in the presence of MNAR-nuclear receptor when compared to genomic activity with test compound in the absence of MNAR and wherein no change is observed in non-genomic activity after addition of said test compound.","33.The method of claim 31, wherein non-genomic activity is increased two-fold after addition of said test compound to a cell in the presence of MNAR-nuclear receptor complex when compared to non-genomic activity with test compound in the absence of MNAR and wherein no change is observed in genomic activity after addition of said test compound.","34.A method of identifying compounds that modulate the non-genomic activity of a nuclear receptor, comprising determining an increase or decrease in the transcriptional activity of a nuclear receptor in the presence of MNAR.","35.A method of determining whether a desired phenotype of cell is effected by non-genomic activity or genomic activity of a nuclear receptor comprising determining the effect of a selected phenotype of cell is increased or decreased when the cell is exposed to additional amounts of MNAR; and then detecting the presence or absence of a desired phenotype, wherein the absence of a desired activity or phenotype, in the presence of additional MNAR indicates increased non-genomic activity of a nuclear receptor negatively affects the desired phenoype (i.e.","the non-genomic activity is not beneficial to said cell) and wherein the increase of a desired phenotype indicates that increased non-genomic activity of a nuclear receptor positively affects said cell.","36.A method according to claims 2, wherein the cell employed is cell that is positively affected by non-genomic activity of the nuclear receptor.","37.A method according to claims 21, wherein the cell employed is cell that is negatively affected by non-genomic activity of the nuclear receptor.","38.A method of identifying compounds that modulate the non-genomic activity of a nuclear receptor, comprising determining an increase or decrease in the transcriptional activity of said nuclear receptor and determining whether a kinase, affected by the presence of MNAR and said nuclear receptor, has an increase or decrease in enzymatic activity.","39.The method of claim 19, wherein the nuclear receptor is a steroid receptor.","40.The method of claim 19, wherein the nuclear receptor is a non-steroid receptor.","41.The method of claim 19, wherein the nuclear receptor is an orphan receptor.","42.The method of claim 19, wherein the steroid receptor is an estrogen receptor.","43.A non-human transgenic mammal, comprising one or more cells which comprise at least one non-functional endogenous MNAR polynucleotide sequence.","44.The non-human transgenic mammal of claim 43, wherein said mammal is selected from the group consisting of a mouse, a rat, a hamster, a guinea pig, a rabbit, a cow, and a sheep.","45.The non-human transgenic mammal of claim 43, comprising at least one non-functional endogenous nuclear receptor.","46.The non-human transgenic mammal of claim 43, wherein the non-functional endogenous receptor is a steroid receptor.","47.The non-human transgenic mammal of claim 43, wherein the non-functional endogenous receptor is a non-steroid receptor.","48.The non-human transgenic mammal of claim 43, wherein the non-functional endogenous receptor is an orphan receptor.","49.The non-human transgenic mammal of claim 43, wherein said the non-functional endogenous receptor is an estrogen receptor.","50.The non-human transgenic mammal of claim 49, wherein endogenous forms of ERα and ERβ are non-functional.","51.The non-human transgenic mammal of claim 43, comprising an interruption of the MNAR coding sequence.","52.The non-human transgenic mammal of claim 49, comprising an interruption of the MNAR and ER coding sequences.","53.The non-human transgenic mammal of claim 43, comprising a nonsense MNAR mutation.","54.The non-human transgenic mammal of claim 49, comprising a nonsense mutations of MNAR and ER.","55.The non-human transgenic mammal of claim 43, comprising a deletion of MNAR coding sequences.","56.The non-human transgenic mammal of claim 43, comprising an alteration in the regulatory region of the MNAR gene making MNAR non-functional.","57.The non-human transgenic mammal of claim 56, comprising substitution of an inducible/repressable promoter for the endogenous MNAR promoter.","58.The non-human transgenic mammal of claim 57, comprising at least one inducible/repressable promoter replacing the endogenous MNAR promoter and the estrogen receptor.","59.The non-human transgenic mammal of claim 43, wherein cells of said mammal further comprise an exogenous selectable marker gene under the control of a promoter active in at least one cell type of said mammal.","60.A method of screening for a compound that disrupts the function of MNAR, comprising using a protein-protein interaction assay to identify compounds that interact with MNAR.","61.A method of screening for a compound that disrupts the function of MNAR, comprising using a two-hybrid assay to identify proteins that interact with MNAR.","62.The method of claim 61 wherein the two-hybrid is conducted in yeast or mammalian cells.","63.A process for producing a polypeptide of claim 3 comprising: (a) culturing a host cell transformed with a nucleic acid sequence for MNAR according to claim 1 under suitable culture medium under conditions for growth; and (b) isolating the polypeptide from the culture medium.","(c) 64.A polypeptide produced from the process of claim 64."],["<SOH> BACKGROUND OF THE INVENTION <EOH>Nuclear hormone receptors are a superfamily of ligand-inducible transcription factors, which, as a class, are involved in ligand-dependent transcriptional control of gene expression.","Binding of a specific ligand, inducing conformational changes in the receptor molecule, affects receptor interaction with other transcription factors, and, ultimately, formation of the preinitiation complex.","This process regulates the rate of gene transcription.","(D. J. Mangelsdorf, et al., Cell 83: 835-9, 1995).","The nuclear hormone receptor superfamily includes steroid hormone receptors, non-steroid hormone receptors, and orphan receptors.","The receptors for glucocorticoids (GR), mineralcorticoids (MR), progestins (PR), androgens (AR), and estrogens (ER) are examples of classical steroid receptors.","In addition to steroid hormone receptors, this nuclear hormone receptor superfamily consists of receptors for non-steroid hormones, such as vitamin D, thyroid hormones, and retinoids.","Moreover, a range of nuclear receptor-like sequences have been identified which encode so called “orphan” receptors.","These orphan receptors are structurally related to and, therefore, classified as nuclear hormone receptors, although no putative ligands have been identified yet.","(Cell, 83: 851-857, 1995).","The superfamily of nuclear hormone receptors is characterized structurally and functionally by a modular structure in comprising six distinct structural and functional domains, A to F. More specifically these receptors have a variable N-terminal region (domain A/B); followed by a centrally located, highly conserved DNA-binding domain (hereinafter referred to as DBD; domain C); a variable hinge region (domain D); a conserved ligand-binding domain (hereinafter referred to as LBD; domain E); and a variable C-terminal region (domain F).","Cell, supra.","The N-terminal region, which is highly variable in size and sequence, is poorly conserved among the different members of the superfamily.","This part of the receptor is involved in the modulation of transcription activation.","The DBD consists of approximately 66 to 70 amino acids and is responsible for DNA-binding activity: This domain targets the receptor to specific DNA sequences, called hormone responsive elements (hereinafter referred to as HRE), within the transcription control unit of specific target genes on the chromatin.","Steroid receptors such as GR, MR, PR, and AR recognize similar HRE DNA sequences, while the ER recognizes a different DNA HRE sequence.","After binding to DNA, the steroid receptor is thought to interact with components of the basal transcriptional machinery and with sequence-specific transcription factors, thereby modulating the expression of specific target genes.","The LBD is located in the C-terminal part of the receptor and is primarily responsible for ligand binding activity.","In this way, the LBD is essential for recognition and binding of the hormone ligand and, in addition, possesses a transcription activation function, thereby determining the specificity and selectivity of the hormone response of the receptor.","Although moderately conserved in structure, the LBD's are known to vary considerably in homology among the individual members of the nuclear hormone receptor superfamily.","When a hormone ligand for a nuclear receptor enters the cell and is recognized by the LBD, it will bind to the specific receptor protein, thereby initiating an allosteric alteration of the receptor protein.","(Cell, supra).","As a result of this alteration the ligand/receptor complex switches to a transcriptionally active state and, as such, is able to bind through the presence of the DBD with high affinity to the corresponding HRE on the chromatin DNA.","In this way, the ligand/receptor complex modulates expression of the specific target genes.","The diversity achieved by this family of receptors results from their ability to respond to different ligands.","In addition to modulation of genomic activity, hormone receptor complexes can have important and varied non-genomic effects.","This non-genomic activity is characterized by fast and transient increase in intracellular second messengers.","In turn, these second messengers are involved in a number of different pathways and cascades that affect cellular functions, such as proliferation and differentiation.","More generally, steroid hormone receptors are connected with embryonic development, adult homeostasis, and organ physiology.","Various diseases and abnormalities are ascribed to a disturbance in steroid hormone action.","Since steroid receptors exercise their influence as hormone-activated transcription modulators and as hormone-activated stimulators of non-genomic activity, further investigation into various approaches to modifying, interacting with, or modulating these receptors is an area of immense significance.","For example, mutations and defects in these receptors, as well as overstimulation or blocking of these receptors, may provide better insight into the underlying mechanism of the hormone signal transduction pathway, thereby leading to increased efficacy in treatment of a wide variety of steroid receptor-linked diseases and abnormalities.","Estrogen Receptors Estrogen (E2) exerts numerous biological effects in different tissues through an interaction with the ER.","Amino acid sequence analyses, transient transfection studies, and mutational dissections of ER indicate that ER has the classical modular structure described above.","The N-terminal A/B domain of the ER contains a transactivation function, referred to as transcriptional activation function 1 (TAF-1).","The DBD contains two zinc fingers and is responsible for DNA recognition.","The LBD and a second transactivation function, referred to as TAF-2, is located at the C-terminal of ER.","Upon binding to hormone, the ER undergoes an activation and transformation step.","The activated ER interacts with specific estrogen response elements (EREs) that are located in the promoter region of estrogen-regulated genes and that influence its target gene transcription.","Over the past decade, numerous studies have provided a basic understanding of both the effects of ligand (agonist/antagonist) on the ER and the relationship between the structure and function of the ER.","Nevertheless, little is known regarding the mechanisms for the non-genomic activity of ER.","ERβ appears to be distinct from the more commonly known estrogen receptor, referred to as ERα.","Collectively, ERα and ERβ are referred to herein as ER.","The DBD of ERβ is 90% identical to that of ERα.","However, the overall homology between the ligand binding domain (LBD) of ERα and ERβ is less than 55%.","Like ERα, ERβ can stimulate transcription from an ERE in a ligand-dependent manner.","It has been established that estrogens induce fast and transient increases in the levels of intracellular second messengers, including calcium and cAMP, and that estrogens induce activation of mitogen-activated protein kinase (MAPK) and phospholipase C (Collins and Webb, 1999).","In fact, numerous studies have demonstrated that estrogens induce rapid and transient activation of the Src/Ras/MAP kinase phosphorylation pathway.","Activation of this pathway triggers vital cellular functions including cell proliferation and differentiation.","The time course of these acute events parallels that elicited by peptide hormones, thus supporting the hypothesis that these events do not involve the ‘classical’ genomic action of estrogens.","Recent data also suggests a direct link between the estrogen receptor and the mitogen-activated protein (MAP) kinase-signaling cascade.","MAP kinases are a family of serine-threonine kinases that are phosphorylated and activated in response to a variety of signals.","These enzymes transduce extracellular signals from multiple membrane receptors to intracellular targets, including transcription factors, cytoskeletal proteins and enzymes.","The MAP kinase family includes the extracellular-signal related kinases (ERKs), p38 and cJun N-terminal kinases which signal through a pathway involving sequential activation of Ras, Raf and mitogen-activated protein kinase (MEK).","(S. M. Thomas, J. S. Brugge, Annual Review of Cell & Developmental Biology 13, 513-609, 1997).","In pulmonary endothelial cells, neuronal cells, osteoblasts and osteoclasts, 17β-estradiol (E2) has been reported to rapidly activate the MAPK pathway.","More specifically, in pulmonary endothelial cells, 17β-estradiol (E2) has been reported to rapidly stimulate nitric oxide (NO) production, thus explaining the ability of E2 to induce acute dilation of blood vessels.","Chen et al.","(Chen et al., 1999) have recently reported that E2 induces rapid activation of endothelial nitric oxide synthase (eNOS) in isolated pulmonary endothelial cells.","Estrogen activation of eNOS was shown to occur through the rapid activation of the MAP kinase pathway.","Complimentary studies have also shown that E2 induces calcium-dependent translocation of eNOS from the plasma membrane to intracellular sites close to the nucleus—an action that is rapid (within 5 minutes), receptor-mediated but nongenomic (Goetz et al., 1999).","In addition, ligand-dependent ERα interaction with the p85α regulatory subunit of phosphatidylinositol-3-OH kinase (PI(3)K) was recently implicated in mediation of cardiovascular protective effects of estrogen (Simoncini T. and J. K., 2000).","Stimulation with estrogen increases ERα associated PI(3)K activity, thereby leading to the activation of protein kinase B/Akt and endothelial nitric oxide synthase (eNOS).","In neuronal cells, rapid activation of the MAP kinase signaling pathway by E2 results in neuroprotection in primary cortical neurons after glutamate excitotoxicity (Singer et al., 1999).","These neuroprotective effects of estrogens, which were reported to occur within 5 minutes after exposure, were mediated through the transient activation of c-Src-tyrosine kinases and tyrosine phosphorylation of p21 (ras)-guanine nucleotide activating protein in an ER-dependent manner.","Similarly, in osteoblasts E2 induces MAP kinase phosphorylation and activation within 5 minutes (Endoh et al., 1997).","MAP kinase activation by estrogens may, therefore, regulate cell proliferation and differentiation in these cells leading to increased bone formation.","Several studies also indicate that estrogens activate the MAP kinase pathway in osteoclasts through activation of Src kinase (Oursler, 1998).","It has been also shown that MAP kinase activation, could be involved in the regulation of enzymes needed for bone resorption (Kristen D. Brubaker, 1999).","In the human mammary cancer derived cell lines, MCF-7 and T47D, as well as in the human colon cancer derived cell line, Caco-2, E2 activates the signal transducing Src/Ras/Erk pathway (Migliaccio et al., 1993), (Migliaccio et al., 1996), (Migliaccio et al., 1998), (Migliaccio et al., 2000).","This activation is mediated by direct ER-Src interaction.","Interestingly, progesterone also activates the same pathway in T47D cells (Migliaccio et al., 1998).","The Src/Ras/ERk signaling pathway is a well-known target of growth factors.","Importantly, activation of this pathway requires direct interaction of ER with Src.","Activation of this pathway triggers different cellular responses such as proliferation or differentiation [Cantley, 1991]; [Marshall, 1996]; [Downward, 1997].","Its activation by estrogen receptor ligands, such as steroid hormones, explicates their involvement in the cell cycle control.","These data support the view that non-transcriptional/nongenomic activity of ER may be responsible for stimulation of cell growth.","However, the details of the molecular mechanism of this process are largely unknown.","Src activation has been previously observed in a large number of human breast and colon carcinomas ([Rosen, 1986]; [Ottenhoff-Kalff, 1992]).","Activation of Src induces mammary tumors in transgenic mice ([Guy, 1994]).","Constitutively active Ras mutants have been found in 25-30% of all cancers, including breast cancer ([Kasid, 1987]).","Recent studies have suggested the existence of a plasma membrane estrogen receptor unrelated to the classical ER.","However, cloning or isolation of this membrane ER has not been accomplished, while others have suggested that a subpopulation of the classical ER is associated with the cell membrane and is responsible for the rapid effects of estrogens.","Transcriptional activity of nuclear receptors itself is a target for tight regulation by multiple signaling pathways, including those stimulated by the neurotransmitter dopamine ([Power, 1991]; [Smith, 1993]), growth factors such as epidermal growth factor (EGF), transforming growth factor-α (TGF-a) and insulin-like growth factor I (IGF-I) ([Ignar-Trowbridge, 1996], [Aronica, 1993]; [Bunone, 1996]), and by activators of protein kinase C ([Aronica, 1994]).","The molecular mechanisms for such cross coupling are thought to be mediated, at least in part, by receptor phosphorylation.","The ER has been demonstrated to be phosphorylated by MAP kinase in response to EGF, thereby resulting in stimulation of AF1 ([Kato, 1996]; [Bunone, 1996]).","Other studies have demonstrated that the ER ([Auricchio, 1987]; [Arnold, 1995]; [Pietras, 1995]), Thyroid hormone receptor (TRβ) ([Lin, 1992]), RARγ ([Rochette-Egly, 1992]), glucocorticoid receptor ([Rao, 1987]) and the orphan receptor HNF-4 ([Ktistaki, 1995]) are also targets for tyrosine phosphorylation.","A specific tyrosine phosphorylation site has been identified within the AF2 domain of the human ER (hER) at amino acid 537 (Y537) ([Castoria, 1993]; [Arnold, 1995]).","This tyrosine is located immediately N-terminal of the AF2 sequence and is conserved in all known ER sequences from diverse species including the ERβ ([Kuiper, 1996]; [Mosselman, 1996]).","Although the molecular mechanisms of non-genomic and genomic activities of E2 are not well understood, the present invention should allow us to create a new generation of therapeutics with tissue and gene selective activity.","To better understand the tissue selective action of some ER ligands, we evaluated the expression and activity of ER-interacting proteins in different cell lines.","Using affinity purification and mass spectrometer based microsequencing, we discovered a novel ER interacting protein, designated as MNAR (modulator of non-genomic activity of nuclear receptors)."],["<SOH> SUMMARY OF THE INVENTION <EOH>The present invention provides for a novel protein modulator of non-genomic activity of nuclear receptors (MNAR) and nucleic acids that encode such proteins.","The MNAR protein can interact with nuclear hormone receptors, such as estrogen receptors.","In general, the protein or polypeptide of MNAR refers to the amino acid sequence of an MNAR.","In particularly preferred embodiments, MNAR protein interacts with ERα and ERβ.","This interaction is ligand dependent.","MNAR will also form a complex with ER and kinases, such as members of the Src family.","Src and MAP kinase activation leads to enhancement of ER transcriptional activity.","The present invention provides (a) a novel nucleic acid comprising a polynucleotide encoding an MNAR having a putative amino acid sequence of SEQ ID No: 2 (see FIG.","12 ).","A nucleotide sequence of human MNAR is shown in SEQ ID No: 1 (See FIG.","15 ).","An amino acid sequence of human MNAR is shown in SEQ ID NO: 13 (See FIG.","16 ).","A nucleotide sequence of murine MNAR is shown in SEQ ID NO: 12 (See FIG.","17 ).","An amino acid sequence of murine MNAR is shown in SEQ ID NO: 13 (See FIG.","18 ).","This invention provides novel isolated nucleic acid comprising a polynucleotide selected from the group consisting of: (a) a polynucleotide encoding a novel protein modulator of non-genomic activity of nuclear receptors (MNAR) comprising the amino acid sequence of SEQ ID NO:2 or of SEQ ID NO:13; (b) a polynucleotide that hybridizes under highly stringent conditions with (i) a region of the nucleotide sequence of SEQ ID NO: 1 or of SEQ ID NO:12, (ii) a subsequence of at least 100 nucleotides of the nucleotide sequence of SEQ ID NO: 1 or of SEQ ID NO:12, (iii) or a complementary strand of (i) or (ii); (c) a polynucleotide comprising a sequence with at least 85% identity to a polynucleotide coding sequence of SEQ ID NO: 1 or of SEQ ID NO: 12; (d) a variant of the polynucleotide comprising a polynucleotide coding sequence of SEQ ID NO:1 or of SEQ ID NO:12; and (e) a polynucleotide encoding a polypeptide fragment comprising the amino acids of SEQ ID NOS 20 or 23 (f) a complement or antisense form of the polynucleotide of (a), (b), (c), (d) or (e); wherein the polynucleotide does not encode a p160 protein ( FIG.","1 ).","In preferred embodiments the isolated nucleic acid comprises an allelic variant of the polynucleotide.","In certain embodiments, the nucleic acid is operably linked to an expression control sequence (e.g.","heterologous or homologous control sequences).","This invention provides for a host cell, including bacteria, yeast, insect, and mammalian cells transformed with such isolated nucleic acids.","This invention also relates to a polypeptides comprising an amino acid sequence selected from the group consisting of: (a) an amino acid sequence for a protein modulator of non-genomic activity of nuclear receptors (MNAR) comprising the amino acid sequence of SEQ ID NO: 2 or of SEQ ID NO:13; (b) an amino acid sequence for an MNAR fragment comprising the amino acid sequence of SEQ ID NOS:14, 15 or 16; (c) a variant of (a) or (b), and (d) a fragment of (a), (b), or (c).","In alternative embodiments, these polypeptides comprise an amino acid sequences which is more than 90%, or 95% identical to the amino acid sequence of SEQ ID NO:13 or 14.In particular embodiments, the polypeptides of the invention comprise the amino acid sequence of SEQ ID NOS 2, 13, 14, 15 16 20 or 22.In additional embodiments the polypeptides of this invention can stimulates the activity of a tyrosine kinase in the presence of one or more nuclear receptors.","The nuclear receptors can be selected from steroid and non-steroid nuclear receptors as well as orphan nuclear receptors.","In certain embodiments, the nuclear receptor is an estrogen receptor.","Alternative embodiments involve monoclonal and polyclonal antibodies which correspond to MNAR polypeptides as well as fragments thereof.","In view of the modulation by MNAR of nuclear receptors, many embodiments of this invention relate to modulating the transcriptional activity of a nuclear receptor, comprising providing to the loci of a nuclear receptor an MNAR polypeptide.","For example, one can modulate the transcriptional activity of a nuclear receptor, by (a) transforming a host cell with the recombinant vector having the nucleic acid sequence for MNAR according to claim 1 ; and (b) culturing said transformed host cell to express the nucleic acid in the presence of a nuclear receptor.","This invention also presents numerous approaches for identifying compounds that affect non-genomic activity or genomic activity of nuclear receptors.","In one embodiment, a method of identifying compounds exhibiting non-genomic activity versus genomic activity compries administering a test compound to a cell comprising an MNAR-nuclear receptor complex and measuring the non-genomic or genomic activity resulting from the test compound.","In these methods, preferably, at least one ligand of a nuclear receptor is present in the cell, or a kinase is present, or both are present.","Other methods are directed to the screening for a compound that modulates the activity of MNAR on a nuclear receptor, comprising the steps of (a) contacting a test compound with an MNAR polypeptide; and (b) determining whether said test compound specifically binds said polypeptide.","In addition, one can conduct a method of screening for a compound that modulates the activity of MNAR on a nuclear receptor, said method comprising the steps of (a) adding a test compound to a cell comprising the polypeptide of and a nuclear receptor; and (b) comparing the MNAR activity before and after said adding step.","Further approaches to this method involve adding a test compound to a control comprising a mutant cell lacking MNAR activity or with significantly reduced MNAR activity.","In each of these methods one can use a control to assess the non-genomic activity (or genomic activity) of a compound by administering a compound to a cell in the presence of MNAR and a nuclear receptor, and then repeating the experiment in the absence of MNAR and comparing the level of non-genomic activity.","Preferred methods employ a cell overexpresses MNAR a nuclear receptor or both.","The genomic activity of a compound is detected by having the nuclear receptor operatively associated with a reporter.","Certain embodiments of the present invention also relate to a method of identifying compounds having selective genomic versus non-genomic activity said method comprising the steps of (a) adding a test compound to a cell comprising a MNAR-nuclear receptor complex; and (b) comparing the genomic versus non-genomic activity before and after said adding the test compound.","A selective genomic activity can be measured by conventional means.","Preferably, the increase or positive effect of non-genomic activity, as measured, is a two-fold increase after addition of said test compound to a cell in the presence of MNAR-nuclear receptor when compared to genomic activity with test compound in the absence of MNAR and wherein no change is observed in non-genomic activity after addition of said test compound.","On the other hand, when selecting for test compounds for non-genomic activity, there should be at least about a two-fold increase after addition of said test compound to a cell in the presence of MNAR-nuclear receptor complex when compared to non-genomic activity with test compound in the absence of MNAR and wherein no change is observed in genomic activity after addition of said test compound.","One can measure the effect by determining an increase or decrease in the transcriptional activity of a nuclear receptor in the presence of MNAR.","In a variety of cells, healthy or diseased, it can be valuable to understand whether a beneficial or healthy state for a cell results from non-genomic or genomic activity.","The present invention permits such determinations by comparing a desired phenotype of a cell, in the presence and absence of MNAR.","The present invention also relates to non-human transgenic mammals, wherein one or more cells comprise at least one non-functional endogenous MNAR polynucleotide sequence, at least one non-functional endogenous nuclear receptor or both.","In preferred embodiments, the non-human transgenic mammal do not have functional endogenous forms of ERa and ERA.","Alternatively, transgenic mammal comprises substitution of an inducible/repressable promoter for the endogenous MNAR promoter.","The non-human transgenic mammal of claim may further comprise an exogenous selectable marker gene under the control of a promoter active in at least one cell type of said mammal.","This invention also relates to a method of screening for a compound that disrupts the function of MNAR, comprising using a protein-protein interaction assay to identify compounds that interact with MNAR.","Once can employ any of the well-known two-hybrid assays or other conventional assays to study protein-protein interactions and the disruption thereof by test compounds, which can be a natural product isolate, small molecule chemical, peptide, etc.","MNAR protein and MNAR encoding nucleic acids can be obtained from mammalian cells and tissues (e.g.","human and murine cells and tissues).","Recombinant MNAR and anti-MNAR antibodies find use in drug screening, diagnostics, and therapeutics.","In particular, the MNAR provides valuable reagents in developing specific biochemical assays for screening compounds for ER ligands that agonize or antagonize the enzymatic activity of the Src family of tyrosine kinases, which are involved in regulating gene expression.","Additionally, the MNAR provides valuable reagents in developing specific biochemical assays for screening compounds for a variety of nuclear receptor ligands.","Accordingly, one can analyze the effects on any pathway involving a nuclear receptor, such as ER, and/or a kinase.","For example, one can use MNAR to set up assays to determine the effect of ER ligands or test compounds on (I) cell cycle stimulation in cancer cells; (II) cell proliferation, cell differentiation and cell conservation for bone development or modulation; (III) neuroprotective activity of ER ligands/test compounds in nerve cells; and (IV) pulmonary characteristics of endothelial cells."],["This application claims the benefit of U.S. application Ser.","No.","60/281,155, filed Apr.","3, 2001.This application is a continuation in part of the aforementioned patent application, the content of which is incorporated by reference.","FIELD OF THE INVENTION The present invention relates to the fields of biochemistry and internal medicine and to a novel modulator of non-genomic activity of nuclear receptors (MNAR), which interacts with both estrogen alpha (ERα) and estrogen beta (ERβ).","More particularly, this invention concerns novel compositions, methods, and assays for modulating the transcriptional activity of estrogen receptors and/or the activation of the src family of tyrosine kinases.","This invention also relates to novel compositions, methods, and assays for differentiating between genomic and non-genomic activity of nuclear receptors (e.g.","estrogen receptors).","Additionally, this invention relates to nucleic acids encoding MNAR, antisense, recombinant expression of nucleic acids, as well as host cells, compositions, and assays that utilize the nucleic acids and protein of this invention.","BACKGROUND OF THE INVENTION Nuclear hormone receptors are a superfamily of ligand-inducible transcription factors, which, as a class, are involved in ligand-dependent transcriptional control of gene expression.","Binding of a specific ligand, inducing conformational changes in the receptor molecule, affects receptor interaction with other transcription factors, and, ultimately, formation of the preinitiation complex.","This process regulates the rate of gene transcription.","(D. J. Mangelsdorf, et al., Cell 83: 835-9, 1995).","The nuclear hormone receptor superfamily includes steroid hormone receptors, non-steroid hormone receptors, and orphan receptors.","The receptors for glucocorticoids (GR), mineralcorticoids (MR), progestins (PR), androgens (AR), and estrogens (ER) are examples of classical steroid receptors.","In addition to steroid hormone receptors, this nuclear hormone receptor superfamily consists of receptors for non-steroid hormones, such as vitamin D, thyroid hormones, and retinoids.","Moreover, a range of nuclear receptor-like sequences have been identified which encode so called “orphan” receptors.","These orphan receptors are structurally related to and, therefore, classified as nuclear hormone receptors, although no putative ligands have been identified yet.","(Cell, 83: 851-857, 1995).","The superfamily of nuclear hormone receptors is characterized structurally and functionally by a modular structure in comprising six distinct structural and functional domains, A to F. More specifically these receptors have a variable N-terminal region (domain A/B); followed by a centrally located, highly conserved DNA-binding domain (hereinafter referred to as DBD; domain C); a variable hinge region (domain D); a conserved ligand-binding domain (hereinafter referred to as LBD; domain E); and a variable C-terminal region (domain F).","Cell, supra.","The N-terminal region, which is highly variable in size and sequence, is poorly conserved among the different members of the superfamily.","This part of the receptor is involved in the modulation of transcription activation.","The DBD consists of approximately 66 to 70 amino acids and is responsible for DNA-binding activity: This domain targets the receptor to specific DNA sequences, called hormone responsive elements (hereinafter referred to as HRE), within the transcription control unit of specific target genes on the chromatin.","Steroid receptors such as GR, MR, PR, and AR recognize similar HRE DNA sequences, while the ER recognizes a different DNA HRE sequence.","After binding to DNA, the steroid receptor is thought to interact with components of the basal transcriptional machinery and with sequence-specific transcription factors, thereby modulating the expression of specific target genes.","The LBD is located in the C-terminal part of the receptor and is primarily responsible for ligand binding activity.","In this way, the LBD is essential for recognition and binding of the hormone ligand and, in addition, possesses a transcription activation function, thereby determining the specificity and selectivity of the hormone response of the receptor.","Although moderately conserved in structure, the LBD's are known to vary considerably in homology among the individual members of the nuclear hormone receptor superfamily.","When a hormone ligand for a nuclear receptor enters the cell and is recognized by the LBD, it will bind to the specific receptor protein, thereby initiating an allosteric alteration of the receptor protein.","(Cell, supra).","As a result of this alteration the ligand/receptor complex switches to a transcriptionally active state and, as such, is able to bind through the presence of the DBD with high affinity to the corresponding HRE on the chromatin DNA.","In this way, the ligand/receptor complex modulates expression of the specific target genes.","The diversity achieved by this family of receptors results from their ability to respond to different ligands.","In addition to modulation of genomic activity, hormone receptor complexes can have important and varied non-genomic effects.","This non-genomic activity is characterized by fast and transient increase in intracellular second messengers.","In turn, these second messengers are involved in a number of different pathways and cascades that affect cellular functions, such as proliferation and differentiation.","More generally, steroid hormone receptors are connected with embryonic development, adult homeostasis, and organ physiology.","Various diseases and abnormalities are ascribed to a disturbance in steroid hormone action.","Since steroid receptors exercise their influence as hormone-activated transcription modulators and as hormone-activated stimulators of non-genomic activity, further investigation into various approaches to modifying, interacting with, or modulating these receptors is an area of immense significance.","For example, mutations and defects in these receptors, as well as overstimulation or blocking of these receptors, may provide better insight into the underlying mechanism of the hormone signal transduction pathway, thereby leading to increased efficacy in treatment of a wide variety of steroid receptor-linked diseases and abnormalities.","Estrogen Receptors Estrogen (E2) exerts numerous biological effects in different tissues through an interaction with the ER.","Amino acid sequence analyses, transient transfection studies, and mutational dissections of ER indicate that ER has the classical modular structure described above.","The N-terminal A/B domain of the ER contains a transactivation function, referred to as transcriptional activation function 1 (TAF-1).","The DBD contains two zinc fingers and is responsible for DNA recognition.","The LBD and a second transactivation function, referred to as TAF-2, is located at the C-terminal of ER.","Upon binding to hormone, the ER undergoes an activation and transformation step.","The activated ER interacts with specific estrogen response elements (EREs) that are located in the promoter region of estrogen-regulated genes and that influence its target gene transcription.","Over the past decade, numerous studies have provided a basic understanding of both the effects of ligand (agonist/antagonist) on the ER and the relationship between the structure and function of the ER.","Nevertheless, little is known regarding the mechanisms for the non-genomic activity of ER.","ERβ appears to be distinct from the more commonly known estrogen receptor, referred to as ERα.","Collectively, ERα and ERβ are referred to herein as ER.","The DBD of ERβ is 90% identical to that of ERα.","However, the overall homology between the ligand binding domain (LBD) of ERα and ERβ is less than 55%.","Like ERα, ERβ can stimulate transcription from an ERE in a ligand-dependent manner.","It has been established that estrogens induce fast and transient increases in the levels of intracellular second messengers, including calcium and cAMP, and that estrogens induce activation of mitogen-activated protein kinase (MAPK) and phospholipase C (Collins and Webb, 1999).","In fact, numerous studies have demonstrated that estrogens induce rapid and transient activation of the Src/Ras/MAP kinase phosphorylation pathway.","Activation of this pathway triggers vital cellular functions including cell proliferation and differentiation.","The time course of these acute events parallels that elicited by peptide hormones, thus supporting the hypothesis that these events do not involve the ‘classical’ genomic action of estrogens.","Recent data also suggests a direct link between the estrogen receptor and the mitogen-activated protein (MAP) kinase-signaling cascade.","MAP kinases are a family of serine-threonine kinases that are phosphorylated and activated in response to a variety of signals.","These enzymes transduce extracellular signals from multiple membrane receptors to intracellular targets, including transcription factors, cytoskeletal proteins and enzymes.","The MAP kinase family includes the extracellular-signal related kinases (ERKs), p38 and cJun N-terminal kinases which signal through a pathway involving sequential activation of Ras, Raf and mitogen-activated protein kinase (MEK).","(S. M. Thomas, J. S. Brugge, Annual Review of Cell & Developmental Biology 13, 513-609, 1997).","In pulmonary endothelial cells, neuronal cells, osteoblasts and osteoclasts, 17β-estradiol (E2) has been reported to rapidly activate the MAPK pathway.","More specifically, in pulmonary endothelial cells, 17β-estradiol (E2) has been reported to rapidly stimulate nitric oxide (NO) production, thus explaining the ability of E2 to induce acute dilation of blood vessels.","Chen et al.","(Chen et al., 1999) have recently reported that E2 induces rapid activation of endothelial nitric oxide synthase (eNOS) in isolated pulmonary endothelial cells.","Estrogen activation of eNOS was shown to occur through the rapid activation of the MAP kinase pathway.","Complimentary studies have also shown that E2 induces calcium-dependent translocation of eNOS from the plasma membrane to intracellular sites close to the nucleus—an action that is rapid (within 5 minutes), receptor-mediated but nongenomic (Goetz et al., 1999).","In addition, ligand-dependent ERα interaction with the p85α regulatory subunit of phosphatidylinositol-3-OH kinase (PI(3)K) was recently implicated in mediation of cardiovascular protective effects of estrogen (Simoncini T. and J. K., 2000).","Stimulation with estrogen increases ERα associated PI(3)K activity, thereby leading to the activation of protein kinase B/Akt and endothelial nitric oxide synthase (eNOS).","In neuronal cells, rapid activation of the MAP kinase signaling pathway by E2 results in neuroprotection in primary cortical neurons after glutamate excitotoxicity (Singer et al., 1999).","These neuroprotective effects of estrogens, which were reported to occur within 5 minutes after exposure, were mediated through the transient activation of c-Src-tyrosine kinases and tyrosine phosphorylation of p21 (ras)-guanine nucleotide activating protein in an ER-dependent manner.","Similarly, in osteoblasts E2 induces MAP kinase phosphorylation and activation within 5 minutes (Endoh et al., 1997).","MAP kinase activation by estrogens may, therefore, regulate cell proliferation and differentiation in these cells leading to increased bone formation.","Several studies also indicate that estrogens activate the MAP kinase pathway in osteoclasts through activation of Src kinase (Oursler, 1998).","It has been also shown that MAP kinase activation, could be involved in the regulation of enzymes needed for bone resorption (Kristen D. Brubaker, 1999).","In the human mammary cancer derived cell lines, MCF-7 and T47D, as well as in the human colon cancer derived cell line, Caco-2, E2 activates the signal transducing Src/Ras/Erk pathway (Migliaccio et al., 1993), (Migliaccio et al., 1996), (Migliaccio et al., 1998), (Migliaccio et al., 2000).","This activation is mediated by direct ER-Src interaction.","Interestingly, progesterone also activates the same pathway in T47D cells (Migliaccio et al., 1998).","The Src/Ras/ERk signaling pathway is a well-known target of growth factors.","Importantly, activation of this pathway requires direct interaction of ER with Src.","Activation of this pathway triggers different cellular responses such as proliferation or differentiation [Cantley, 1991]; [Marshall, 1996]; [Downward, 1997].","Its activation by estrogen receptor ligands, such as steroid hormones, explicates their involvement in the cell cycle control.","These data support the view that non-transcriptional/nongenomic activity of ER may be responsible for stimulation of cell growth.","However, the details of the molecular mechanism of this process are largely unknown.","Src activation has been previously observed in a large number of human breast and colon carcinomas ([Rosen, 1986]; [Ottenhoff-Kalff, 1992]).","Activation of Src induces mammary tumors in transgenic mice ([Guy, 1994]).","Constitutively active Ras mutants have been found in 25-30% of all cancers, including breast cancer ([Kasid, 1987]).","Recent studies have suggested the existence of a plasma membrane estrogen receptor unrelated to the classical ER.","However, cloning or isolation of this membrane ER has not been accomplished, while others have suggested that a subpopulation of the classical ER is associated with the cell membrane and is responsible for the rapid effects of estrogens.","Transcriptional activity of nuclear receptors itself is a target for tight regulation by multiple signaling pathways, including those stimulated by the neurotransmitter dopamine ([Power, 1991]; [Smith, 1993]), growth factors such as epidermal growth factor (EGF), transforming growth factor-α (TGF-a) and insulin-like growth factor I (IGF-I) ([Ignar-Trowbridge, 1996], [Aronica, 1993]; [Bunone, 1996]), and by activators of protein kinase C ([Aronica, 1994]).","The molecular mechanisms for such cross coupling are thought to be mediated, at least in part, by receptor phosphorylation.","The ER has been demonstrated to be phosphorylated by MAP kinase in response to EGF, thereby resulting in stimulation of AF1 ([Kato, 1996]; [Bunone, 1996]).","Other studies have demonstrated that the ER ([Auricchio, 1987]; [Arnold, 1995]; [Pietras, 1995]), Thyroid hormone receptor (TRβ) ([Lin, 1992]), RARγ ([Rochette-Egly, 1992]), glucocorticoid receptor ([Rao, 1987]) and the orphan receptor HNF-4 ([Ktistaki, 1995]) are also targets for tyrosine phosphorylation.","A specific tyrosine phosphorylation site has been identified within the AF2 domain of the human ER (hER) at amino acid 537 (Y537) ([Castoria, 1993]; [Arnold, 1995]).","This tyrosine is located immediately N-terminal of the AF2 sequence and is conserved in all known ER sequences from diverse species including the ERβ ([Kuiper, 1996]; [Mosselman, 1996]).","Although the molecular mechanisms of non-genomic and genomic activities of E2 are not well understood, the present invention should allow us to create a new generation of therapeutics with tissue and gene selective activity.","To better understand the tissue selective action of some ER ligands, we evaluated the expression and activity of ER-interacting proteins in different cell lines.","Using affinity purification and mass spectrometer based microsequencing, we discovered a novel ER interacting protein, designated as MNAR (modulator of non-genomic activity of nuclear receptors).","SUMMARY OF THE INVENTION The present invention provides for a novel protein modulator of non-genomic activity of nuclear receptors (MNAR) and nucleic acids that encode such proteins.","The MNAR protein can interact with nuclear hormone receptors, such as estrogen receptors.","In general, the protein or polypeptide of MNAR refers to the amino acid sequence of an MNAR.","In particularly preferred embodiments, MNAR protein interacts with ERα and ERβ.","This interaction is ligand dependent.","MNAR will also form a complex with ER and kinases, such as members of the Src family.","Src and MAP kinase activation leads to enhancement of ER transcriptional activity.","The present invention provides (a) a novel nucleic acid comprising a polynucleotide encoding an MNAR having a putative amino acid sequence of SEQ ID No: 2 (see FIG.","12).","A nucleotide sequence of human MNAR is shown in SEQ ID No: 1 (See FIG.","15).","An amino acid sequence of human MNAR is shown in SEQ ID NO: 13 (See FIG.","16).","A nucleotide sequence of murine MNAR is shown in SEQ ID NO: 12 (See FIG.","17).","An amino acid sequence of murine MNAR is shown in SEQ ID NO: 13 (See FIG.","18).","This invention provides novel isolated nucleic acid comprising a polynucleotide selected from the group consisting of: (a) a polynucleotide encoding a novel protein modulator of non-genomic activity of nuclear receptors (MNAR) comprising the amino acid sequence of SEQ ID NO:2 or of SEQ ID NO:13; (b) a polynucleotide that hybridizes under highly stringent conditions with (i) a region of the nucleotide sequence of SEQ ID NO: 1 or of SEQ ID NO:12, (ii) a subsequence of at least 100 nucleotides of the nucleotide sequence of SEQ ID NO: 1 or of SEQ ID NO:12, (iii) or a complementary strand of (i) or (ii); (c) a polynucleotide comprising a sequence with at least 85% identity to a polynucleotide coding sequence of SEQ ID NO: 1 or of SEQ ID NO: 12; (d) a variant of the polynucleotide comprising a polynucleotide coding sequence of SEQ ID NO:1 or of SEQ ID NO:12; and (e) a polynucleotide encoding a polypeptide fragment comprising the amino acids of SEQ ID NOS 20 or 23 (f) a complement or antisense form of the polynucleotide of (a), (b), (c), (d) or (e); wherein the polynucleotide does not encode a p160 protein (FIG.","1).","In preferred embodiments the isolated nucleic acid comprises an allelic variant of the polynucleotide.","In certain embodiments, the nucleic acid is operably linked to an expression control sequence (e.g.","heterologous or homologous control sequences).","This invention provides for a host cell, including bacteria, yeast, insect, and mammalian cells transformed with such isolated nucleic acids.","This invention also relates to a polypeptides comprising an amino acid sequence selected from the group consisting of: (a) an amino acid sequence for a protein modulator of non-genomic activity of nuclear receptors (MNAR) comprising the amino acid sequence of SEQ ID NO: 2 or of SEQ ID NO:13; (b) an amino acid sequence for an MNAR fragment comprising the amino acid sequence of SEQ ID NOS:14, 15 or 16; (c) a variant of (a) or (b), and (d) a fragment of (a), (b), or (c).","In alternative embodiments, these polypeptides comprise an amino acid sequences which is more than 90%, or 95% identical to the amino acid sequence of SEQ ID NO:13 or 14.In particular embodiments, the polypeptides of the invention comprise the amino acid sequence of SEQ ID NOS 2, 13, 14, 15 16 20 or 22.In additional embodiments the polypeptides of this invention can stimulates the activity of a tyrosine kinase in the presence of one or more nuclear receptors.","The nuclear receptors can be selected from steroid and non-steroid nuclear receptors as well as orphan nuclear receptors.","In certain embodiments, the nuclear receptor is an estrogen receptor.","Alternative embodiments involve monoclonal and polyclonal antibodies which correspond to MNAR polypeptides as well as fragments thereof.","In view of the modulation by MNAR of nuclear receptors, many embodiments of this invention relate to modulating the transcriptional activity of a nuclear receptor, comprising providing to the loci of a nuclear receptor an MNAR polypeptide.","For example, one can modulate the transcriptional activity of a nuclear receptor, by (a) transforming a host cell with the recombinant vector having the nucleic acid sequence for MNAR according to claim 1; and (b) culturing said transformed host cell to express the nucleic acid in the presence of a nuclear receptor.","This invention also presents numerous approaches for identifying compounds that affect non-genomic activity or genomic activity of nuclear receptors.","In one embodiment, a method of identifying compounds exhibiting non-genomic activity versus genomic activity compries administering a test compound to a cell comprising an MNAR-nuclear receptor complex and measuring the non-genomic or genomic activity resulting from the test compound.","In these methods, preferably, at least one ligand of a nuclear receptor is present in the cell, or a kinase is present, or both are present.","Other methods are directed to the screening for a compound that modulates the activity of MNAR on a nuclear receptor, comprising the steps of (a) contacting a test compound with an MNAR polypeptide; and (b) determining whether said test compound specifically binds said polypeptide.","In addition, one can conduct a method of screening for a compound that modulates the activity of MNAR on a nuclear receptor, said method comprising the steps of (a) adding a test compound to a cell comprising the polypeptide of and a nuclear receptor; and (b) comparing the MNAR activity before and after said adding step.","Further approaches to this method involve adding a test compound to a control comprising a mutant cell lacking MNAR activity or with significantly reduced MNAR activity.","In each of these methods one can use a control to assess the non-genomic activity (or genomic activity) of a compound by administering a compound to a cell in the presence of MNAR and a nuclear receptor, and then repeating the experiment in the absence of MNAR and comparing the level of non-genomic activity.","Preferred methods employ a cell overexpresses MNAR a nuclear receptor or both.","The genomic activity of a compound is detected by having the nuclear receptor operatively associated with a reporter.","Certain embodiments of the present invention also relate to a method of identifying compounds having selective genomic versus non-genomic activity said method comprising the steps of (a) adding a test compound to a cell comprising a MNAR-nuclear receptor complex; and (b) comparing the genomic versus non-genomic activity before and after said adding the test compound.","A selective genomic activity can be measured by conventional means.","Preferably, the increase or positive effect of non-genomic activity, as measured, is a two-fold increase after addition of said test compound to a cell in the presence of MNAR-nuclear receptor when compared to genomic activity with test compound in the absence of MNAR and wherein no change is observed in non-genomic activity after addition of said test compound.","On the other hand, when selecting for test compounds for non-genomic activity, there should be at least about a two-fold increase after addition of said test compound to a cell in the presence of MNAR-nuclear receptor complex when compared to non-genomic activity with test compound in the absence of MNAR and wherein no change is observed in genomic activity after addition of said test compound.","One can measure the effect by determining an increase or decrease in the transcriptional activity of a nuclear receptor in the presence of MNAR.","In a variety of cells, healthy or diseased, it can be valuable to understand whether a beneficial or healthy state for a cell results from non-genomic or genomic activity.","The present invention permits such determinations by comparing a desired phenotype of a cell, in the presence and absence of MNAR.","The present invention also relates to non-human transgenic mammals, wherein one or more cells comprise at least one non-functional endogenous MNAR polynucleotide sequence, at least one non-functional endogenous nuclear receptor or both.","In preferred embodiments, the non-human transgenic mammal do not have functional endogenous forms of ERa and ERA.","Alternatively, transgenic mammal comprises substitution of an inducible/repressable promoter for the endogenous MNAR promoter.","The non-human transgenic mammal of claim may further comprise an exogenous selectable marker gene under the control of a promoter active in at least one cell type of said mammal.","This invention also relates to a method of screening for a compound that disrupts the function of MNAR, comprising using a protein-protein interaction assay to identify compounds that interact with MNAR.","Once can employ any of the well-known two-hybrid assays or other conventional assays to study protein-protein interactions and the disruption thereof by test compounds, which can be a natural product isolate, small molecule chemical, peptide, etc.","MNAR protein and MNAR encoding nucleic acids can be obtained from mammalian cells and tissues (e.g.","human and murine cells and tissues).","Recombinant MNAR and anti-MNAR antibodies find use in drug screening, diagnostics, and therapeutics.","In particular, the MNAR provides valuable reagents in developing specific biochemical assays for screening compounds for ER ligands that agonize or antagonize the enzymatic activity of the Src family of tyrosine kinases, which are involved in regulating gene expression.","Additionally, the MNAR provides valuable reagents in developing specific biochemical assays for screening compounds for a variety of nuclear receptor ligands.","Accordingly, one can analyze the effects on any pathway involving a nuclear receptor, such as ER, and/or a kinase.","For example, one can use MNAR to set up assays to determine the effect of ER ligands or test compounds on (I) cell cycle stimulation in cancer cells; (II) cell proliferation, cell differentiation and cell conservation for bone development or modulation; (III) neuroprotective activity of ER ligands/test compounds in nerve cells; and (IV) pulmonary characteristics of endothelial cells.","BRIEF DESCRIPTION OF THE DRAWINGS FIG.","1 is an alignment comparing the amino acid sequence of MNAR and p160, a protein homologous to MNAR.","FIG.","2 shows estrogen receptor β interaction with MNAR.","Panel A presents a silver stained gel obtained by GST-ERβ-LBD pull-down from MCF-7 cells extract.","The resulting peptides were purified using reverse phase HPLC column and identified by mass spectrometry based microsequencing.","The sequences of these peptides are provided in panel B.","FIG.","3.depicts the results of the expression analysis of MNAR in various cell lines, as evaluated by Northern blotting from experiments in Example 2.FIG.","4 (A and B) shows the results of MNAR expression analysis in various tissues, as evaluated by Northern blotting from experiments in Example 2.FIGS.","5A and 5B show analysis of flag-tagged MNAR interaction with ERα and ERβ, or receptors liganded with E2, or 4-OH tamoxifen.","Also shown are the results of full-length flag-MNAR interaction with transcribed/translated androgen (AR) and glucocorticoid receptors (GR) with and without their corresponding ligands (FIGS.","5C and 5D).","FIG.","6A depicts the results of the experiment of example 4, wherein the tests are conducted to evaluate the in vitro interaction of Src or Lck with MNAR and/or ERa in the presence and in the absence of 17β-estradiol.","FIG.","6B depicts the results of the experiment of example 4, wherein the tests are conducted to evaluate the in vitro interaction of Src or Lck with MNAR and/or ERβ in the presence and in the absence of 17β-estradiol.","FIG.","7 is a schematic depiction of the proposed model of ER-MNAR-Src interaction.","FIG.","8 is an SDS-PAGE gel analysis of the ER-MNARc complex on cSrc enzymatic activity.","cSrc enzymatic activity was evaluated in the absence (lanes 1 and 2) or presence of ER (lanes 3-6), E2 (lanes 3 and 5), and MNAR (lanes 2, 5, and 6) using acid denatured enolase as a substrate.","Data presented in FIG.","8 indicate that MNAR itself strongly stimulates cSrc enzymatic activity.","FIG.","9 is a series of graphs depicting the results from the experiments of example 7, wherein the tests are conducted to evaluate whether MNAR affects ER-mediated transcription.","FIG.","9A shows the result for ERα, MNAR, and/or SRC3.FIG.","9B depicts the results for ERα and MNAR when cells are treated with E2, E2 plus PP2, or E2 plus PD98059.FIG.","9C shows results of MCF-7 cells treated with antisense (AS), E2, E2 plus control, or E2 plus antisense.","A primer/probe set is designed to target MNAR, PS2, and cathepsin D, respectively.","FIG.","9D shows results of a Western blot analysis of MCF-7 cell extracts transfected with antisense or reverse control oligomers stimulated with E2 with rabbit polyclonal MNAR antiserum.","FIG.","10 depicts an SDS-gel analysis of MCF-7 cells untransfected and/or transfected with a Flag-MNAR expression vector untreated and treated with ±2.Flag-MNAR (panel A), ERα (panel C and D), and cSrc (panel B) were immunoprecipitated from the cell extracts using their corresponding antibodies.","Material precipitated using ERα antiserum was probed with rabbit polyclonal anti-MNAR antibody (panel E).","E2-enhancement of ERα-MNAR interaction was clearly detected.","FIG.","11 depicts the results of the analysis of MNAR on E2 induced Erk activation, as evaluated by Western blotting from experiments in Example 9.FIG.","12 is a schematic depiction of the structural-functional organization of MNAR and 13 MNAR mutants (mutants 2-14), which were generated and tested for their ability to enhance ER-stimulated transcription when transfected into HepG2 cells under the experiments of Example 10.FIG.","13 is a bar graph depicting the results of an analysis of MNAR deletion mutants (mutants 2-14), which were tested for their ability to enhance ER-stimulated transcription when transfected into hepG2 cells in the experiments of Example 10.FIG.","14 depicts is a bar graph depicting the results of an analysis of PXXP motif MNAR mutants, which were tested for their ability to activate ER transcriptional activity When transfected into HepG2 cells in the experiments of Example 10.FIG.","15 depicts a polynucleotide sequence of human MNAR (SEQ ID NO:1).","FIG.","16 depicts an amino acid sequence of human MNAR (SEQ ID NO: 2).","FIG.","17 depicts a polynucleotide sequence of murine MNAR (SEQ ID NO:12)).","FIG.","18 depicts an amino acid sequence of murine MNAR (SEQ ID NO:13).","DESCRIPTION OF THE SPECIFIC EMBODIMENTS Definitions Functional analogs of a given MNAR protein or protein with “substantial functional identity” to a given MNAR are defined as proteins that exhibit one or more biochemical properties specific to such MNAR.","“Nucleic acid molecule” means a single-stranded or double stranded molecule and includes RNA molecules, DNA molecules, and analogs of RNA or DNA molecules.","“Modulating transcription” means altering transcription, and includes changing the rate of transcription initiation, the level of transcription, or the responsiveness of transcription/transcription initiation to regulatory controls.","“Neoplastic state of cells” means any new growth of cells which may be benign or malignant.","The terms “substantially pure” or “isolated” mean that the MNAR protein, MNAR protein fragment, or nucleic acid encoding a MNAR or MNAR fragment are unaccompanied by at least some of the material with which it is normally associated in its natural state.","A composition of a substantially pure MNAR or portion thereof may contain excipients and additives useful in diagnostic, therapeutic and investigative reagents.","“Altered expression state” means that the expression of MNAR in a naturally-occurring state is altered to increase or decrease MNAR expression.","In comparison to the naturally-occurring state.","“Substantial sequence identity” means that a portion of the protein or nucleic acid presents at least about 70%, more preferably at least about 80%, and most preferably at least about 90%, 95%, or 99% sequence identity with a MNAR sequence portion.","Where the sequence diverges from native MNAR sequences disclosed herein, the differences are preferably conservative, i.e., an acidic for an acidic amino acid substitution or a nucleotide change providing a redundant codon.","Dissimilar sequences are typically aggregated within regions rather than being distributed evenly over the polymer.","A substantially identical sequence hybridizes to a complementary MNAR-encoding sequence under relatively high stringency.","For the purposes of defining high stringency southern hybridization conditions, reference can conveniently be made to Sambrook et al.","(1989) at pp.","387-389 which is herein incorporated by reference where the washing step at paragraph 11 is considered high stringency.","“Fusion protein or polypeptide” means a protein or polypeptide that comprises at least a first amino acid sequence fused to a second amino acid sequence, where such fusion does not occur in a naturally occurring protein or polypeptide.","Preferably, at least one amino acid sequence in the fusion protein or polypeptide is that of MNAR.","“Mutant or mutation” means any detectable change in genetic material, e.g.","DNA, or any process, mechanism or result of such a change.","This includes gene mutations, in which the structure (e.g.","DNA sequence) of a gene is altered, any gene or DNA arising from any mutation process, and any expression product (e.g.","protein, polypeptide, or enzyme) expressed by a modified gene or DNA sequence.","“Variant” is used to indicate a modified or altered gene, DNA sequence, protein, polypeptide, enzyme, cell, etc., i.e.","any kind of mutant.","A variant of a naturally occurring protein or polypeptide, for example, comprises a substitution, deletion, modification, and/or insertion of one or more amino acids in the naturally occurring protein or polypeptide.","“Recombinant” means any genetic material (e.g.","DNA sequence) resulting from the insertion into the sequence or chain, by biological or chemical means, of a sequence (e.g.","a full-length or partial DNA sequence) not originally biologically or naturally present in that genetic material (e.g.","DNA sequence).","“Protein or polypeptide fragment” means any smaller portion of a larger protein or polypeptide, where preferably the fragment has at least 8 amino acids and more preferably, 25, 50, 100 or 200 amino acids.","Non-genomic activity of a receptor is “beneficial to a cell” wherein the cell exhibits presence of a desired activity or phenotype upon introduction of increased amount of MNAR to the cell.","Non-genomic activity of a receptor is “not beneficial to a cell” wherein the cell exhibits absence of a desired activity or phenotype upon introduction of increased amount of MNAR to the cell.","MNAR Characterization Sequence analysis MNAR revealed multiple LXXLL motifs localized in N-terminal portion of MNAR′ molecule.","Similar motifs in other transcription factors were previously shown to form contacts with a hydrophobic groove on the surface of the ligand binding domains of nuclear receptors ((Heery et al., 1997), (Torchia et al., 1997)).","Another important feature of the MNAR molecule is a proline-rich domain.","This domain can be utilized for interaction with SH3 domains of the Src family of protein kinases.","Initially we hypothesized that MNAR may potentiate nuclear receptors interaction with Src kinases.","Our data indicate that estrogen receptors α and β interact with MNAR, and this interaction is enhanced by 17β-estradiol but not by 4(OH) tamoxifen.","Interestingly, the MNAR-ER complex, both in-vitro and in-vivo strongly interacts and stimulates enzymatic activity of Src kinases.","Importantly, overexpression of MNAR leads to enhancement of ER transcriptional activity.","We expect that this activation of ER is due to its phosphorylation by one of the kinases downstream of Src.","Accordingly, we submit that MNAR modulates ER interaction with Src kinases and by doing so mediates the nongenomic activity of ER.","Our data supports the development of experimental models that will allow us to assay for ER ligands that are able to differentiate between genomic and non-genomic activity of ER ligands.","Ligands that do not affect transcriptional activity of ER, but stimulate the Src enzymatic activity may possess important bone-sparing, CNS protective, and cardioprotective activity.","On the other hand, compounds that selectively control transcriptional activity of ER would have a much more selective action, thereby avoiding multiple side affects.","Overall, our ability to separate genomic and nongenomic activity of nuclear receptor ligands, particularly ER ligands, may allow us to create a new generation of therapeutics with profoundly focused, selective action.","Our data also support the development of experimental models that will allow us to assay for nuclear receptor ligands that are able to differentiate between genomic and non-genomic activity of nuclear receptor ligands.","Ligands that do not affect transcriptional activity of various nuclear receptors, but stimulate the Src enzymatic activity or other non-genomic enzymatic activity may possess important therapeutic properties or activities.","On the other hand, compounds that selectively control transcriptional activity of nuclear receptors would have a much more selective action, thereby avoiding multiple side affects.","Overall, our ability to separate genomic and non-genomic activity of nuclear receptor ligands may allow us to create a new generation of therapeutics with profoundly focused, selective action.","Nucleic Acid Sequences Encoding MNAR and Corresponding MNAR Proteins A biologically active MNAR, or MNAR fragment thereof, retains one or more of the MNAR's functions, such as the ability to specifically form a complex with ER, or to modulate or enhance the transcriptional activity of ER.","Exemplary assays for biological activity are described below and in the working exemplification.","Specific binding is empirically determined by contacting, for example a MNAR, with a mixture of components and identifying those components that preferentially bind the MNAR.","Specific binding may be conveniently shown by a number of methods, including but not limited to yeast and mammalian hybrid systems and competitive binding studies.","For example, a human cDNA library was screened using the yeast two-hybrid assay to identify potential coactivators, receptors and/or ligands for ER or MNAR.","In addition to the analysis of the interactions between kinases and the MNAR-ER complex, MNAR can be used in similar strategies to assess the interaction between ER ligands and ER with other known transcription factors, including other nuclear receptors, and to identify novel interacting proteins.","For example, since the MNAR-ER interaction can be enhanced by 17β-estradiol, one can assess the effect of test agents, which may be compounds or proteins or peptides.","Furthermore, the yeast two-hybrid system, the mammalian two-hybrid system, surface plasmon resonance assays, immunoprecipitation assays, or other assays can be used to demonstrate a direct interaction between MNAR and ER in the presence or absence of Src.","Similarly, these assay systems could be used to broadly screen for known and novel proteins that interact with MNAR-ER complex or MNAR-ER-Src complex.","The invention provides recombinantly produced MNAR proteins, MNAR analogs and fragments thereof.","These recombinant products are readily modified through physical, chemical, and molecular techniques disclosed or cited herein or otherwise known to those skilled in the relevant art.","According to a particular embodiment of the invention, fragments of the MNAR-encoding sequences are spliced with heterologous sequences to produce fusion proteins.","Such fusion proteins find particular use in modulating ER activity in vitro and in vivo.","MNAR can be further modified by methods known in the art.","For example, MNAR may be phosphorylated or dephosphorylated, glycosylated or deglycosylated, with or without radioactive labeling, etc.","Serine, threonine, and tyrosine residues can particularly provide useful phosphorylation sites.","Especially useful are modifications that alter MNAR solubility, membrane transportability, stability, and binding specificity and affinity.","Some examples include fatty acid-acylation, proteolysis, and mutations in nuclear receptor transcription factor interaction domains that stabilize binding.","Particularly, one may modify the proline-rich region of one of the LXXLL motifs.","A substantially pure or isolated MNAR protein, or MNAR portion encoded by nucleic acid, is generally at least about 1% homologous to said MNAR encoding nucleic acid; preferably at least about 10%; more preferably at least about 50%; and most preferably at least 90%.","Nucleic acid weight percentages are determined by dividing the weight of the MNAR or MNAR portion encoding nucleic acid, including alternative forms and analogs such as alternatively spliced or partially transcribed forms, by the total nucleic acid weight present.","The invention also provides for MNAR sequences modified by transitions, transversions, deletions, insertions, or other modifications such as alternative splicing and such alternative forms, genomic MNAR sequences, MNAR gene flanking sequences, including MNAR regulatory sequences and other non-transcribed MNAR sequences, MNAR mRNA sequences, and RNA and DNA antisense sequences complementary to MNAR encoding sequences, sequences encoding xenogeneic MNARs and MNAR sequences comprising synthetic nucleotides, e.g., the oxygen of phosphate group may be replaced with sulfur, methyl, or the like.","The modified MNAR encoding sequences or related sequences encode proteins with MNAR-like functions.","There will generally be substantial sequence identity between at least a portion thereof and a portion of a MNAR, preferably at least about 40%, more preferably at least 80%, most preferably at least 90%, 95%, or 99%, particularly conservative substitutions, particularly within the proline-rich regions and LXXLL motifs and regions encoding protein domains involved in protein-protein interactions, particularly MNAR-ER or MNAR-ER-kinase interactions, particularly MNAR-nuclear receptor or MNAR-nuclear receptor-kinase interactions.","MNAR can be subject to alternative purification, biosynthesis, modification or use by methods disclosed herein or otherwise known in the art.","For example, the amino acids can be modified in a number of ways, including but not limited to, altering stability, solubility, binding affinity, specificity, and methylation.","The amino acid sequences of the present invention may also be modified with a label capable of providing a detectable signal, either directly or indirectly.","Exemplary labels include, but are not limited to, radioisotopes, fluorescers, and biotinylation.","Nucleic acids encoding at least a portion of an MNAR are used to identify nuclear factors and agents which interact with that MNAR using expression screening in yeast or mammalian cells as described in Current Protocols in Molecular Biology (See “Protein Expression” in Chapter 16, 2002).","One method of identifying these nuclear factors and agents, which interact with MNAR, is to utilize a mammalian, or yeast, two-hybrid or three-hybrid assay.","(See, e.g., Fields S. et al., (1994) Trends Genet.","10:286-292; Young P. et al.","(1992) Current Biology 3:408-420; Zervos et al.","(1993) Cell 72:223-232; Madura et al.","(1993) J. Biol.","Chem.","268:12046-12054; Bartel et al (1993) Biotechniques 14:920-924; and Iwabuchi et al.","(1993) Oncogene 8:1693-1696).","As exemplified herein, a yeast cDNA library containing fusion genes of cDNA joined with DNA encoding the activation domain of a transcription factor (e.g.","Gal4) are transfected with fusion genes encoding a portion of a MNAR and the DNA binding domain of a transcription factor.","Clones encoding MNAR binding proteins provide for the complementation of the transcription factor and are identified through transcription of a reporter gene.","See, e.g.","Fields and Song (1989) Nature 340, 245-246 and Chien et al.","(1991) Proc.","Natl.","Acad.","Sci USA 88, 9578-9582.The invention also provides vectors comprising nucleic acids encoding MNAR or portion or analog thereof and optionally, encoding ERs and or kinases.","A large number of vectors, including plasmid and viral vectors, have been described for expression in a variety of eukaryotic and prokaryotic hosts.","Vectors will often include one or more replication systems for cloning or expression, one or more markers for selection in the host, e.g., antibiotic resistance, and one or more expression cassettes.","The inserted MNAR coding sequences may be synthesized, isolated form natural sources, prepared as hybrids, etc.","Ligation of the coding sequences to the transcriptional regulatory sequences may be achieved by known methods.","Advantageously, vectors may also include a promoter operably linked to the MNAR encoding portion.","Suitable host cells may be transformed/transfected/infected by any suitable method including electroporation, CaCl2 mediated DNA uptake, viral infection, microinjection, microprojectile, or other established methods.","Alternatively, nucleic acids encoding one or more MNARs may be introduced into cells by recombination events.","For example, a sequence can be microinjected into a cell, and thereby effect homologous recombination at the site of an endogenous gene encoding a MNAR, an analog or pseudogene thereof, or a sequence with substantial identity to a MNAR-encoding gene.","Other recombination based methods such as non-homologous recombinations, deletion of endogenous gene by homologous recombination, especially in pluripotent cells, etc., provide additional applications.","Appropriate host cells include bacteria, archebacteria, fungi, especially yeast, and plant and animal cells, especially mammalian cells.","Of particular interest are E. coli, B. subtilis, Saccharomyces cerevisiae, A549 cells, CHO, COS, HeLa, Hep2, and bone cells and immortalized mammalian myeloid and lymphoid cell lines.","Preferred host cell lines include but are not limited to human cancer cell lines such as MCF-7 and T47D cells.","Ideally, such expression systems utilize inducible expression strategies like the TET ON/OFF system that is commercially available.","Such cell lines are useful to define the role of MNAR in non-genomic ER activity and to assess the effect of potential compounds that modulate MNAR-ER interactions.","A large number of transcription initiation and termination regulatory elements/regions have been isolated and shown to be effective in the transcription and translation of heterologous proteins in the various hosts.","Examples of these regions, methods of isolation, manner of manipulation, etc.","are known in the art.","Suitable expression control sequences and host cell/cloning vehicle combinations are well known, and are described in Sambrook et al.","(1989).","MNAR encoding oligonucleotides can also be used to identify other MNARs or transcription factor coactivators.","For example, 32P-labeled MNAR encoding nucleic acids are used to screen cDNA libraries at low stringency to identify similar cDNAs that encode proteins with MNAR related domains.","Additionally, MNAR related proteins are isolated by PCR amplification with degenerate oligonucleotide probes using the sequences disclosed herein.","Other experimental methods for cloning MNAR are also set out in the working exemplification below.","Other useful cloning, expression, and genetic manipulation techniques for practicing the inventions disclosed herein are known to those skilled in the art.","The compositions and methods disclosed herein may be used to effect gene therapy.","See, e.g., Gutierrez et al.","(1992) Lancet 339, 715-721.For example, cells are transfected with MNAR sequences operably linked to gene regulatory sequences capable of effecting altered MNAR expression or regulation.","To modulate MNAR translation, cells may be transfected with MNAR complementary antisense polynucleotide.","Antisense modulation may employ MNAR antisense sequences operably linked to gene regulatory sequences.","An antisense oligonucleotide can be, for example, 8, 10, 15, 20, 25, 30, 35, 40, 45 or 50 nucleotide sequences in length.","An antisense nucleic acid can be constructed using chemical synthesis and enzymatic ligation reactions using known procedures in the art.","For example, an antisense nucleic acid (e.g.","antisense oligonucleotide) can be chemically synthesized using naturally occurring nucleotides or various modified nucleotides designed to increase the biological activity of the molecules or to increase the physical stability of the duplex formed between antisense and sense nucleic acids, e.g.","phosphorothiolate derivatives and acridine substituted nucleotides can be used.","Alternatively, the antisense nucleic acid can be produced biologically using an expression vector into which a nucleic acid has been subcloned in an antisense orientation (i.e.","RNA transcribed from the inserted nucleic acid will be of an antisense orientation to a target nucleic acid of interest.","In the present invention, cells are transfected with a vector comprising a MNAR sequence with a promoter sequence oriented such that transcription of the gene yields an antisense transcript capable of binding to MNAR encoding mRNA.","Transcription may be constitutive or inducible and the vector may provide for stable extrachromosomal maintenance or integration.","Alternatively, single-stranded antisense nucleic acid sequences that bind to genomic DNA or mRNA encoding at least a portion of MNAR may be administered to the target cell at a concentration that results in a substantial reduction in MNAR expression.","Assays for Identifying Modulators of Non-Genomic Activity of Nuclear Receptors and Therapeutic Agents The invention provides methods and compositions for identifying agents useful in modulating the genomic or non-genomic activity of nuclear receptors (e.g.","ER).","Such agents find use in the diagnosis or treatment of broad range of disease including, but not limited to, cancer, cardiovascular diseases, microbial and fungal infections and particularly immune disease, bone protection, etc.","The ability to develop rapid and convenient high-throughput biochemical assays for screening compounds that work through nuclear receptor (e.g.","ER) non-genomic activity versus nuclear receptor (e.g.","ER) genomic activity in human cells opens a new avenue for drug development.","In addition, this invention provides a tool for identifying whether disease states within a cell or beneficial cell states of a cell are effected by nuclear receptor (e.g.","ER) genomic/non-genomic activities/pathways, i.e.","one can determine whether cancer cells proliferate when nuclear receptor (e.g.","ER) non-genomic activity or nuclear receptor (e.g.","ER) genomic activity is decreased/enhanced.","In general, this invention provides for categorizing many cell types as requiring nuclear receptor (e.g.","ER) genomic/non-genomic activity.","Common assays include yeast two-hybrid assays as disclosed by Young and Ozenberger in U.S. Pat.","No.","5,989,808, which is herein incorporated by reference.","The two hybrid assays can be used to screen for small molecule compounds or peptides or proteins of interest.","Typically, prospective agents are screened from large libraries of synthetic or natural compounds.","For example, numerous means are available for random and directed synthesis of saccharide, peptide, and nucleic acid based compounds, see, e.g.","Lam et al.","(1991) Nature 354, 82-86.Alternatively, libraries and compounds are readily modified through conventional chemical, physical, and biochemical means.","Examples of such modifications are disclosed herein.","Useful agents are identified with a range of assays employing MNARs or MNAR encoding nucleic acids.","The assays may also employ one or more selected from nuclear receptors, steroid receptors, orphan receptors, ER, ERE (estrogen response elements), ER ligands, kinases and inhibitors of such kinases.","As examples, protein binding assays, nucleic acid binding assays and gel shift assays are useful approaches.","Exemplary assays include assaying labeled nuclear receptor (e.g.","ER) binding to immobilized MNAR, labeled MNAR or MNAR peptide binding to immobilized nuclear receptor (e.g.","ER), etc.","Many appropriate assays are amenable to scaled-up, high throughput usage suitable for volume drug screening.","The particular assay used will be determined by the particular nature of the MNAR interactions.","Assays may employ a single MNAR, MNAR fragments, MNAR fusion products, partial MNAR complexes, or the complete basal transcription complex comprising an MNAR nucleic acid.","Useful agents are typically those that bind to or modify the association of MNAR, the MNAR-nuclear receptor complex, or the MNAR-ER complex.","Preferred agents include those capable of modulating the expression of MNAR genes, particularly genes transcribed by members of the nuclear receptor superfamily.","Useful agents are found within numerous chemical classes, though typically they are organic compounds; preferably small organic compounds.","Small organic compounds have a molecular weight of more than 50 yet less than about 2,500, preferably less than about 750, more preferably, less than about 250.Exemplary classes include peptides, saccharides, steroids, and the like.","Selected agents may be modified to enhance efficacy, stability, pharmaceutical compatibility, and the like.","Structural identification of an agent may be used to identify, generate, or screen additional agents.","For example, where peptide agents are identified, they may be modified in a variety of ways to enhance their stability, such a using an unnatural amino acid, such as a D-amino acid, particularly D-alanine, by functionalizing the amino or carboxyl terminus, e.g., for the amino group, acylation or alkylation, and for the carboxyl group, esterification or amidification, or the like.","Other methods of stabilization may include encapsulation, for example, in liposomes, etc.","Agents may be prepared in a variety of ways known to those skilled in the art.","For example, peptides under about 60 amino acids can be readily synthesized today using conventional commercially available automatic synthesizers.","Alternatively, peptide (and protein and nucleic acid agents) are readily produced by known recombinant technologies.","These assay systems could also be used to screen for potential therapeutics, including peptide and chemical ligands.","Such potential therapeutics, activators or inhibitors of MNAR function, could act by either modulating an interaction between MNAR and either a nuclear receptor (e.g.","ER) or a kinase (“class I”) or by modulating an activity of MNAR or a nuclear receptor (e.g.","ER) or kinase (“class II”).","Interaction assays, including but not limited to two-hybrid assays, immunoprecipitation assays, and surface plasmon resonance (SPR) assays, could be used to identify “class I” therapeutics.","For example, chemical compounds or peptides can be screened for their ability to modulate the interaction between MNAR and a nuclear receptor (e.g.","ER) by SPR.","Activity assays, including but not limited to mammalian transfection assays in which nuclear receptor (e.g.","ER) transcriptional activity is observed, could be used to identify “class II” compounds.","For example, compounds could be screened for their ability to modulate the enhancement of nuclear receptor (e.g.","ER) activity by MNAR.","Such compounds may modulate the interactions of MNAR with a nuclear receptor (e.g.","ER) or they may modulate a known or unknown activity of MNAR or a nuclear receptor (e.g.","ER), including a transcriptional activity or assays that can be used to evaluate enzymatic activity of kinases.","For therapeutic uses, the compositions and selected agents disclosed herein may be administered by any convenient way that will depend upon the nature of the compound.","For small molecular weight agents, oral administration is preferred and enteric coatings may be indicated where the compound is not expected to retain after exposure to the stomach environment.","Generally the amount administered will be empirically determined by routine dose-response experiments.","Typically amounts range from about 1 to 1000 μg/kg of recipient.","Large proteins are preferably administered parenterally, conveniently in a physiologically acceptable carrier, e.g., phosphate buffered saline, saline, deionized water, or the like.","Typically, such compositions are added to a retained physiological fluid such as blood or synovial fluid.","Other additives may be included, such as stabilizers, bactericides, etc.","These additives will be present in conventional amounts.","MNAR may also be modified with a label capable of providing a detectable signal either directly or indirectly.","Exemplary labels include radioisotopes, fluorescers, etc.","Alternatively, a MNAR may be expressed in the presence of a labeled amino acid such as 35S-methionine.","Such labeled MNAR and analogs thereof find use, for example, as probes in expression screening assays for proteins that interact with MNAR, or, for example, MNAR binding to ER or other ER interacting proteins or transcription factors in drug screening assays.","Specific polyclonal or monoclonal antibodies that can distinguish MNAR from other nuclear proteins are readily made using the methods and compositions disclosed in Harlow and Lane, Antibodies, A Laboratory Manual, Cold Spring Harbor Laboratory, 1988, other references cited herein, as well as immunological and hybridoma technologies known to those in the art.","In particular, MNAR and analogs and fragments thereof also find use in raising anti-MNAR antibodies in laboratory animals such as mice and rabbits as well as the production of monoclonal antibodies by cell fusion or transformation.","Anti-MNAR antibodies and fragments (Fab, etc.)","thereof find use in modulating MNAR role in transcription complexes, screening MNAR expression libraries, etc.","In addition, these antibodies can be used to identify, isolate, and purify structural analogs of MNAR.","Anti-MNAR antibodies also find use for subcellular localization of MNAR under various conditions such as infection, during various cell cycle phases, induction with cytokines, protein kinases such as C and A, etc.","Other exemplary applications include using MNAR-specific antibodies (including monoclonal or MNAR-derived peptide specific antibodies) to immuno-deplete in vitro transcription extracts and using immuno-affinity chromatography to purify MNAR, including analogs, or other nuclear factors which interact with MNAR.","Compositions are also provided for therapeutic intervention in disease, for example, by modifying MNARs or MNAR encoding nucleic acids.","Oligopeptides can be synthesized in pure form and can find many uses in diagnosis and therapy.","These polypeptides can be used, for example, to modulate native MNAR interaction with MNAR interacting proteins.","The oligopeptides will generally be more than six and fewer than about 60 amino acids, more usually fewer than about 30 amino acids, although large oligopeptides may be employed.","A MNAR or a portion thereof may be used in purified form, generally greater than about 50%, usually greater than about 90% pure.","Methods for purifying such peptides to such purities include various forms of chromatographic, chemical, and electrophoretic separations disclosed herein or otherwise known to those skilled in the art.","Transgenic Animals and Cells Animal models serve as useful vehicles for screening of compounds, for example, for effects on cholesterol levels, cholesterol absorption, and bile acid synthesis.","Transgenic animals, defined as animals with engineered genomes, may prove particularly useful in certain applications.","In the present application, the focus is on the modulation of ER activity, or other nuclear receptors and the role of MNAR in effecting the non-genomic activity of ER.","Thus, embodiments of this invention include transgenic mice that lack MNAR activity as well as transgenic animals that may lack nuclear receptor activity, ER activity, or a kinase activity.","Animals lacking one of the above are useful to determine the specificity of a compound or ligand for MNAR, ER, a kinase.","These animals are useful not only in elucidating the biology of this signaling pathway, but as tools in examining possible therapeutic compositions and methods of treatment.","For these reasons, transgenic non-human animals of the present invention will have at least one non-functional endogenous MNAR (and/or ER and/or kinase).","It further is contemplated that the transgenic non-human animal of the present invention may have both MNAR and ER altered, thus providing cells which lack functional MNAR-ER complex.","The non-functional attribute(s) can contain an interruption of the gene coding sequence, a nonsense mutation that truncates the protein product, a deletion of the gene coding sequence or alterations in the MNAR regulatory region.","In designing a heterologous gene for expression in animals, sequences which interfere with the efficacy of gene expression, such as coding sequences, promoters, introns, polyadenylation signals, polymerase II termination sequences, hairpins, consensus splice sites and the like, may be eliminated.","Current advances in transgenic approaches and techniques have permitted the manipulation of a variety of animal genomes via gene addition, gene deletion, or gene modifications (Franz et al., 1997).","The transgenic synthesis of human hemoglobin (U.S. Pat.","No.","5,602,306, specifically incorporated herein by reference) and fibrinogen (U.S. Pat.","No.","5,639,940, specifically incorporated herein by reference) in non-human animals have also been disclosed, each specifically incorporated herein by reference in it entirety.","The construction of a transgenic mouse model has recently been used to assay potential treatments for Alzheimer's disease (U.S. Pat.","No.","5,720,936, specifically incorporated herein by reference in its entirety).","It is contemplated in the present invention that transgenic animals contribute valuable information as models for studying the effects of ligands specific for the MNAR and/or ER in transgenic MNAR and ER knockout animal models.","Conclusions This invention is based on the molecular cloning, structural analysis and characterization of a protein modulator of non-genomic activity of nuclear receptors (MNAR).","The present invention demonstrates that human MNAR interacted with nuclear hormone receptors to form a complex and affect the enzymatic activity of Src protein kinases.","Crosstalk between cell surface form of ER and intracellular signaling pathways was first demonstrated more than 20 years ago (Pietras, 1975), (Pietras, 1977).","Since then multiple reports confirmed rapid estrogen effects occurring in the vasculature, breast, bone, uterus and neuronal tissues that cannot be explained by the “genomic” action of estrogen receptor.","It became clear that biological activity of estrogen in responsive cells is mediated not only through a specific high-affinity receptor, located exclusively in their nuclei, but also through the interaction of estrogen with cell-surface receptor.","The nature of this receptor is debated.","Some evidence supports the existence of a unique membrane receptor, other favors the identity or the strong similarity between the cell-surface and nuclear estrogen receptors (Pietras and Szego, 1979), (Bression et al., 1986), (Pappas et al., 1995).","Important evidence, based on experiments with COS cells transfected with ERa cDNA, clearly demonstrated that classical estrogen receptor can activate cSrc in the process leading to activation of tyrosine phosphorylation/p21ras/MAP kinase pathway (Migliaccio et al., 1996).","Evaluating the spectrum of ER interacting proteins in MCF7 cells, we discovered a novel ER-interacting protein—MNAR.","We showed that both ERα and ERβ interacted with MNAR and affinity of this interaction is enhanced by 17β-estradiol and reduced by 4(OH) tamoxifen (FIG.","3).","Protein homologous to MNAR—p160 was recently isolated using pull-down approach with SH2 domain of Lck.","Our data indicate that MNAR is the major form of this protein expressed in MCF7 cells (FIG.","2).","Next we evaluated whether MNAR affects estrogen receptors interaction with Src family kinases.","Our data indicate that both ERα and ERβ interact with partially purified cSrc and this interaction is not enhanced by 17β-estradiol (FIGS.","4A and B).","This result does not contradict previously reported data (Migliaccio et al., 2000), (Migliaccio et al., 1998) since we used in vitro transcribed/translated ER and MNAR and also purified cSrc.","At the same time Migliaccio et al.","(Migliaccio et al., 2000), (Migliaccio et al., 1998) performed their experiment by pulling down Src- or ER-interacting proteins from cell extracts that may contain MNAR.","Importantly, in presence of MNAR, estrogen receptors interact with cSrc and Lck in ligand-dependent way (FIGS.","4A and B).","We also observed that MNAR itself binds to cSrc or Lck with low affinity.","Our results therefore suggest that ER and MNAR synergistically bind to Src family kinases.","Under basal conditions, the catalytic domain of Src family kinases is constrained in an inactive state through intermolecular interactions.","Binding of the SH2 domain to the C-terminal phosphorylated tyrosine locks the molecule in an inhibited conformation ([Matsuda, 1990]).","Binding of the SH3 domain to the proline-reach domain of the linker region is also important for kinase inactivation ([Superti-Furga, 1993]).","Full catalytic activation requires release of these restrains.","The kinase activity of Src can be enhanced by binding of the SH2 domain to phosphotyrosine-containing sequences and by binding of the SH3 domain to the proline-rich sequences ([Hubbard, 1998]).","We next investigated if ER-MNAR binding leads to cSrc activation.","Our results suggest that MNAR strongly enhances cSrc catalytic activity that can be further stimulated by addition of ER-E2 (FIG.","5).","Phosphotyrosine 537 of ERα has been identified as the residue essential for ER interaction with SH2 domain of Src ((Migliaccio et al., 1998)).","We submit that ER interacts with LXXLL motifs of the MNAR molecule and ER's phosphotyrosine 537 interacts with SH2 domain of Src.","At the same time proline-rich domain of MNAR probably interacts with SH3 domain of Src.","Formation of this complex leads to cSrc activation.","Activation of cSrc in response to EGF has been previously shown to promote the ER phosphorylation by the MAP kinase and stimulation of ER-mediated transcription ([Kato, 1996]; [Bunone, 1996]).","Our results indicate that overexpression of MNAR, presumably through activation of cSrc-mediated phosphorylation cascade, augments ER transcriptional activity (FIG.","6A).","Attenuation of MNAR-induced ER activation in the presence of Src inhibitors PP2 and genistein (FIG.","6B) supports the view that MNAR interacts with ER via the cSrc-mediated phosphorylation cascade.","Our studies describe a new ER-interacting protein that modulates the cross talk between ER and Src family tyrosine kinases, thus regulating the nongenomic activity of estrogens and potentially other steroids.","The nongenomic activity represent an important mechanism by which estrogens regulates cell grows and proliferation.","MNAR therefore, may represent an important target for development of new estrogens with more selective action.","The above conclusions are supported by the following examples, which are offered by way of illustration and not by way of limitation.","EXAMPLES Equipment and Reagents.","17β-estradiol (E2) and 4(OH) tamoxifen were obtained from Sigma.","ICI-182,780 was provided by AstraZeneca Pharmaceuticals (Wlimington.","DE).","5′RACE primers were from marathon cDNA kit (BD Biosciences Clontech, Palo Alto, Calif.).","Purified cSrc or Lck were from Upstate (Charlottesville, Va.).","Biotinylated peptide derived from p34cdc2 was synthesized by the Peptide Chemistry group at Wyeth Research.","Glutathione agarose beads were obtained from Sigma-Aldrich (St. Louis, Mo.).","Anti-phosphotyrosine antibody and SuperSignal Elisa Pico peroxidase substrate were from Perbio Science AB (Pierce, Rockford, Ill.).","Src inhibitor PP2 was from Calbiochem-Novobiochem Corporation (San Diego, Calif.).","Method 1: Isolation and identification of MNAR.","GST-ERβ-LBD and GST-ERα-LBD were expressed in BL21 (DE3) E. coli (GST is Glutathione S-Transferase).","Cell culture in LB at OD0.6 was induced with 0.1 mM IPTG, incubated for 3 hr at 25° C. and cells were pelleted by centrifugation.","Bacterial cells were sonicated 5 ml of 20 mM Tris pH 7.2, 500 mM NaCl, 1 mM DTT, 0.2 mM EDTA, 0.1 mM PMSF, centrifuged at 40,000 g for 30 min.","To immobilize GST-ERβ-LBD and GST-ERα-LBD cell extract was incubated with 50% glutathione agarose slurry in 20 mM Tris pH 7.2, 180 mM NaCl, 1 mM DTT, 0.005% NP40 (Binding Buffer) at 4° C. MCF7 cell extract was prepared using NE-PER Nuclear and Cytoplasmic Extraction Reagents (Perbio Science AB, Pierce, Rockford, Ill.) according to manufacture instructions.","0.5 mg of MCF7 cell extract was incubated at 4° C. with either GST or GST-ER-LBD fusion protein with or without 17-β-estradiol (0.1 μM) at 4° C. Bound material was boiled off the agarose beads in 2× SDS buffer and purified on 10% PAA Gel.","The gel was silver stained.","Method 2: Mass Spectrometric Analysis of Purified Proteins.","Protein samples purified by electrophoresis were excised manually, then reduced, alkylated and digested with trypsin (Promega, Madison, Wis.) in situ using an automated in-gel tryptic digestion robot ((Houthaeve T, 1997)).","After digestion, peptide extracts were concentrated to a final volume of 10-20 μl using a Savant Speed Vac Concentrator (ThermoQuest, Holdbrook, N.Y.).","Peptide extracts were analyzed on an automated microelectrospray reversed phase HPLC.","In brief, the microelectrospray interface consisted of a Picofrit fused silica spray needle, 50 cm length by 75 um ID, 8 um orifice diameter (New Objective, Cambridge Mass.)","packed with 10 um C18 reversed-phase beads (YMC, Wilmington, N.C.) to a length of 10 cm.","The Picofrit needle was mounted in a fiber optic holder (Melles Griot, Irvine, Calif.) held on a home-built base positioned at the front of the mass spectrometer detector.","The rear of the column was plumbed through a titanium union to supply an electrical connection for the electrospray interface.","The union was connected with a length of fused silica capillary (FSC) tubing to a FAMOS autosampler (LC-Packings, San Francisco, Calif.) that was connected to an HPLC solvent pump (ABI 140C, Perkin-Elmer, Norwalk, Conn.).","The HPLC solvent pump delivered a flow of 50 uL/min which was reduced to 250 nL/min using a PEEK microtight splitting tee (Upchurch Scientific, Oak Harbor, Wash.), and then delivered to the autosampler using an FSC transfer line.","The LC pump and auto sampler were each controlled using their internal user programs.","Samples were inserted into plastic autosampler vials, sealed, and injected using a 5 ul sample loop.","Extracted peptides from the surface digests were concentrated 10-fold using a Savant Speed Vac Concentrator (ThermoQuest, Holdbrook, N.Y.), and then were separated by the microelectrospray HPLC system using a 50 minute gradient of 0-50% solvent B (A: 0.1M HoAc, B: 90% MeCN/0.1M HoAc).","Peptide analyses were done on a Finnigan LCQ-DECA ion trap mass spectrometer (ThermoQuest, San Jose, Calif.) operating at a spray voltage of 1.5 kV, and using a heated capillary temperature of 125° C. Data were acquired in automated MS/MS mode using the data acquisition software provided with the instrument.","The acquisition method included 1 MS scan (375-600 m/z) followed by MS scans of the top 2 most abundant ions in the MS scan.","The instrument then did a second MS scan (600-1000 m/z) followed by MS/MS scans of the top 2 most abundant ions in that scan.","The dynamic exclusion and isotope exclusion functions were employed to increase the number of peptide ions that were analyzed (settings: 3 amu=exclusion width, 3 min=exclusion duration, 30 secs=pre-exclusion duration, 3 amu=isotope exclusion width).","Automated analysis of MS/MS data was performed using the SEQUEST computer algorithm ((Houthaeve T, 1997)) incorporated into the Finnigan Bioworks data analysis package (ThermoQuest, San Jose, Calif.) using the non-redundant (NR) database of proteins obtained from the NCBI genome center.","Method 3: MNAR Cloning: Three pairs of oligonucleotides were designed and used to clone the N-terminal, middle, and C-terminal portions of MNAR Oligo #1: TAGGATCCAGATGGCGGCAGCCGTTCTGAG (SEQ ID NO 3) and oligo #2: CGATCAGGATCCCAAAGC (SEQ ID NO 4) were used to clone the N-terminal portion.","Oligo #3: GCTTTGGGATCCTGATCG (SEQ ID NO 5) and oligo #4: CAAGGAGATCTCCACATC (SEQ ID NO 6) were used to clone the middle portion.","Oligo #5: GATGTGGAGATCTCCTTG (SEQ ID NO 7) and oligo #6: GCTAGGAGTCAGGCTCTG (SEQ ID NO 8) were used to clone the C-terminal portion.","1×107 MCF7 cells were lyzed in Trizol (Life Technologies, Rockville, Md.)","for total RNA isolation according to protocol.","40 ng of total RNA and 400 nM of oligos was used in a RT-PCR reaction using One Step RT-PCR kit (Qiagen, Valencia, Calif.) to amplify MNAR starting with 30 min at 50° C., followed by 15 min at 95° C., and then 30 cycles of 30 sec at 95° C., 30 sec at 55° C., 60 sec at 72° C., and extension at 72° C. for 7 min.","The respective products were cloning into pT-Adv vector (Invitrogen, Carlsbad, Calif.) and sequenced.","Full length MNAR was assembled by restriction enzyme digestion and ligation.","MNAR was also independently cloned by using oligo #1 and #6 from a human lymphoma marathon cDNA library according to protocol (BD Biosciences Clontech, Palo Alto, Calif.).","The MNAR cloned from MCF-7 was identical to the MNAR cloned from marathon cDNA library.","A 5′RACE was performed using oligo #7 CCGMGCCAAGACACACAGTGCTGCTGGMTAG (SEQ ID NO 9) and adaptor primer 1 oligo form marathon cDNA kit (BD Biosciences Clontech, Palo Alto, Calif.) to obtain additional sequence information.","Method 4: MNAR Expression Analysis.","Total RNA (10 ug) from HepG2, SaOS2, HOBs 02/02, COS-7, MCF-7.T47D, MDA231, Ishikawa, 1-10, LNCaPLN3, LNCaPFGC, JCA, TSUPRL cells were prepared by Trizol, separated on gel, transferred to membrane, and probed with radiolabeled oligo #7 corresponding to nucleotide 367 to 400 of cloned MNAR.","Another oligo probe #8 CTGGAGAAAAAAGGGGCAGAGATAAAGAGT (SEQ ID NO 10), corresponding to nucleotide 1480 to 1509 of deposited Genbank sequence, was used to probe MCF-7 RNA in another Northern.","A second MNAR expression analysis was performed.","Radiolabeled oligonucleotide probe corresponding to nucleotide 367 to 393 of MNAR was hybridized to a human multi-tissue northern blot II (BD Biosciences Clontech, Palo Alto, Calif.) in Perfect Hybridization buffer (Sigma-Aldrich, St. Louis, Mo.)","at 42° C. The blot was washed three times in 0.2×SSC, 0.1% SDS at 42° C. and exposed to film.","β-Actin probe was used as a control.","Method 5: Transient Transfection Analysis: HepG2 and COS-7 cells were routinely maintained in DMEM supplemented with 10% FBS.","Either 50,000 cells per well of HepG2 or 30,000 per well of COS-7 were seeded in 96 well plates in DMEM without phenol red supplemented with 10% charcoal stripped serum.","100 ng of 2×ERE-tk-Luc reporter, 10 ng of pCMV-O-gal internal control, either 1 ng of pcDNA3.1 ERα or 5 ng of pcDNA3.1 ERβ expression vector with or without increasing amount of pcDNA3.1 MNAR or pcDNA3.1 SRC3 expression plasmids were transfected using 0.5 ul of lipofectamine 2000 (Life Technologies, Rockville, Md.)","according to protocol.","Cells were stimulated either with 10−8M 17-b-estradiol, tamoxifen, or ICI for 24 hr before being processed for luciferase and β-Galactosidase activity.","Method 6: MNAR-ER Interaction Analysis: MNAR-Nuclear receptors interaction analysis was performed.","SF9 cells extract, overexpressing full-length flag-MNAR, were incubated with in vitro transcribed/translated [35S]-labeled full-length ERα (FIG.","5A), ERβ (FIG.","5B), androgen (AR) (FIG.","5C) and glucocorticoid receptors (GR) (FIG.","5D) with and without their corresponding ligands, all at 1 μM.","Formed complexes were isolated using anti-FLAG sepharose beads, boiled in 2×SDS buffer and loaded on SDS gel.","The gel was dried and subjected to autoradiography.","Method 7: ER-MNAR-cSrc Interaction Analysis.","In vitro transcribed/translated ERα with (FIG.","6A, lanes 4, 5 and 8, 9) or without MNAR (FIG.","6A, lanes 2, 3 and 6, 7) in presence (FIG.","6A, lanes 3, 5, 7 and 9) or absence of 1 μM E2 (FIG.","6A, lanes 2, 4, 6 and 8), were incubated with purified cSrc (FIG.","6A, lanes 2-5) or Lck (FIG.","6A, lanes 6-9).","Formed complexes were isolate by pulled-down with anti-cSrc or anti-Lck antibodies and protein A agarose, boiled in 2×SDS buffer and resolved on an SDS-PAGE gel.","Gel was dried and subjected to autoradiography.","Method 8: Analysis of cSrc Enzymatic Activity.","The cSrc tyrosine kinase activity was evaluated in the absence (FIG.","8, lanes 1 and 2) or presence of ER (FIG.","8, lanes 3-6), 17β-estradiol (FIG.","8, lanes 3 and 5), and MNAR (FIG.","8, lanes 2, 5 and 6) using acidified enolase as a substrate.","Before reaction proteins were incubated for 1 hr at 4° C. Phosporylation reaction was started by addition of [32P]-γATP and continued at 30° C. for 10 min.","Reaction was stopped by addition of 2×SDS buffer.","Phosphorylated proteins were separated by SDS-PAGE and the dried gel was subjected to autoradiography.","Method 9: MNAR Effect on ER-Mediated Transcription.","A—HepG2 cells were transfected with expression plasmids for ERα, MNAR and/or SRC3.Luciferase gene expression, driven by a 2×ERE-tk-reporter, was evaluated in cells treated with E2, 4(OH)-tamoxifen, or ICI 182,780 all at 10 nM.","B—HepG2 cells were transfected with expression vectors for ERαand MNAR.","Luciferase gene expressions, driven by a 2xERE-tk-reporter, were evaluated in cells treated with 10 nM E2, 10 nM E2 plus 1 μM PP2, or 1 μM PD98059.C—MCF-7 cells were transfected with 100 nM antisense (AS) CATGGAGATGTCCCGGAACAGTGCA (SEQ ID NO: 11) from Sequitur, Inc. (Natick, Mass.)","or reverse control (C) oligomers complexed with 1 μg/ml of Lipofectamine 2000 in white media.","After 24 hours cells were stimulated with 10 nM E2, 24 hours later cells were lysed in guanidium buffer and total RNA was isolated from lysates.","TaqMan analysis was performed using a primer/probe set designed to target MNAR, pS2 and Cathepsin D. D—Extracts of MCF7 cells transfected with 100 nM antisense (AS)—1, or reverse control (C)—2 oligomers stimulated with 10 nM E2 were used for Western Blotting Analysis with rabbit polyclonal MNAR antiserum.","MNAR antiserum was generated against 11-mer peptide encoding amino acids 509-520 (SHRKGDSNANSD) (SEQ ID NO: 31) of the MNAR.","Method 10: MNAR and ER Interaction and Activation of Src Kinase MNAR and ERα interact with Src and promote phosphorylation of endogenous proteins.","MCF-7 cells transfected or not transfected with FLAG-MNAR expression vector were not treated (FIG.","10, lanes 1 and 3) or treated with E2 at 10 nM for 5 minutes (lanes 2 and 4).","FLAG-MNAR (panel A), cSrc (panel B), or ERα (panels C, D and E) were immunoprecipitated from the cell extracts using corresponding antibodies.","Immunoprecipitates were incubated with [32P]γATP for 30 minutes at 30° C. (panels A-C), or with [32P]-γATP and enolase (panel D).","Kinase reaction was stopped by boiling in 2×SDS buffer.","Phosphorylated material was loaded on SDS gel, which was then dried and subjected to autoradiography.","Material obtained by ERα pull down from cells not transfected with FLAG-MNAR, untreated (1) and treated with E2 (2) was probed with MNAR-antiserum (panel E).","Method 11: MNAR affect on E2 Induced Erk Kinase 1 and 2 Activation It has been previously reported that activation of Src by estradiol triggers Ras/Erks kinase pathway.","We expected that MNAR induced Src activation may lead to activation of Erk kinases 1 and 2.We evaluated the level of Erk activation by E2 in MCF7 cells, cells overexpressing MNAR, and cells in which MNAR expression was substantially attenuated by treatment with MNAR antisense oligonucleotide (SEQ ID NO:11).","Antibodies that recognize phosphorylated Erk 1 and 2 were used to detect E2 induced stimulation of Erk 1 and 2 phosphorylation.","Extracts of MCF7 cells (FIG.","11, lanes 3 and 4) transfected with MNAR expression plasmid (FIG.","11, lanes 1 and 2), or MNAR antisense oligonucleotide at 100 nM (FIG.","11, lanes 5 and 6); unstimulated (FIG.","11, lanes 1, 3 and 5), or stimulated for 5 minutes (FIG.","11, lanes 2, 4 and 6) with 10 nM E2 were used for Western blotting analysis with antibodies against phosphorylated Erk 1 and 2—panel A; Erk1/2 antibodies—panel B; MNAR antibodies—panel C. MNAR antisense oligonucleotide (SEQ ID NO:11) was developed at Sequitur Inc. (Natick, Mass.).","Method 12: MNAR Structural-Functional Analysis MNAR sequence analysis revealed that the N-terminal portion of its molecule contains multiple LXXLL motifs previously found to be important for interaction with nuclear hormone receptors, while at the C-terminus, MNAR contains a region rich in both proline and glutamic acid.","To delineate the regions of MNAR that are responsible for interaction with estrogen receptor and c-Src tyrosine kinase, we performed functional evaluation of MNAR deletion mutants (FIG.","12).","A series of C-terminal truncations of MNAR was generated using specially designed oligonucleotides, by amplifying appropriate fragments and cloning them into appropriate expression vectors.","These mutants were tested for their abilities to enhance ER-stimulated transcription when transfected into HepG2 cells.","Method 13: Screening for Compounds that Affect Kinase Activity.","Compounds are screened as follows.","The reaction buffer used is 50 mM Tris-HCl, pH 7.0, 50 mM MgCl2, 50 uM Na3VO4.The substrate peptide is obtained and 6 mg dissolve in 1.7 mL of RB and store at −20° C. ATP to be used is at 400 uM final concentration.","An inhibitor is to be used, for example, Piceatannol—3 mM (stored at 4° C. and protected from light).","First, ER incubation with E2.For example, 10 μl of commercially available ER (Panvera) is incubated with 40 μl of RB and 0.5 μl of E2 at 10−6M.","The mix is incubated at RT for 1 hour.","Next, the mixture is placed in the presence or absence of MNAR, E2, and ER.","The solutions are incubated at RT for 30 minutes.","To each tube, 10 μl ATP (final concentration 400 μM) is added along with 10 μl substrate peptide.","This is incubated for 60 minutes at 30° C. The reaction is stopped by adding 10 μl of 3 mM piceatannol.","Now the solution is incubated at RT for 5 minutes.","Dilution and peptide immobilization is performed.","10 μl Reaction is diluted with 20 ml H2O.","200 μl of dilution is added to wells of NeutrAvidin Plate Washing is performed by adding 200 μl of PBS 5× for 3 minutes each.","This is followed by addition of Anti-P-Tyr antibody.","The antibody is diluted in PBS and BSA and the solution is covered and allowed to incubate at 37° C. for one hour.","The mixture is washed the same as above.","Then a substrate reaction is generated by adding 100 μl of Pierce substrate and reading in Victor using the HRP protocol.","Experimental Analysis and Results Example 1 MNAR Isolation and Identification, Cloning of cDNA Encoding MNAR.","To better understand the tissue-selective action of ER ligands, we established a proteomics approach to evaluate the expression and activity of ER-interacting proteins in different cell lines.","We identified MNAR using GST-pull down approach.","A GST-ERβ ligand-binding domain (LBD) was incubated with a MCF-7 cell extract.","FIG.","2A presents a silver-stained gel of the fractions obtained by the pull-down experiment.","A number of proteins were detected that specifically interacted with GST-ERβ-E2, but not with GST-ERβ, or GST alone.","Corresponding bands were excised, digested with trypsin, concentrated, and separated by the microelectrospray HPLC system.","Peptide analysis was done on a Finnigan LCQ-DECA ion trap mass spectrometer (ThermoQuest, San Jose, Calif.).","Automated analysis of mass spectrometry data was performed using the SEQUEST computer algorithm ([Houthaeve T, 1997]) incorporated into the Finnigan Bioworks data analysis package (ThermoQuest, San Jose, Calif.) using the non-redundant database of proteins obtained from the NCBI genome center.","FIG.","2A shows a silver-stained gel resulting from the experimental steps as outlined above in Method 1.Cell extract was incubated with Glutathione beads—1, bacterially expressed GST-ERβ-LBD fusion protein in the absence of ligand—2, or presence of E2—3, or 4(OH) tamoxifen—4, both at 10−7M.","ER-interacting proteins were isolated using anti-GST agarose and separated on SDS-PAGE.","The gel area containing the corresponding band was excised and subjected to trypsin digestion.","The bands corresponding to the proteins revealing ligand-dependent interaction were excised, digested with trypsin and identified using mass spectrometry based peptide microsequencing.","Together with known and well-characterized coactivators, such as SRC3 and DRIP205, we identified a protein that had not been previously shown to interact with ERs.","This protein interacted with the GST-ERβ-LBD in the presence of E2 and had an apparent molecular weight of approximately 120 kDa (FIG.","2A).","Peptide sequence analysis revealed that this protein could be related to a proline and glutamic acid rich protein—-p160 (Genbank U88153), previously isolated by pull-down with the Src homology domain 2 (SH2) of p56lck (Lck) (21).","FIG.","2B presents sequences of the peptides identified by the mass spectrometry and their position in the MNAR molecule.","Searching the NCI database we also identified several ESTs that matched this protein.","One of them, AL03939, matched at the N-terminus and extended the 5′ sequence an additional 100 bp.","The aligned sequence was used to design oligonucleotide primers to clone this protein from MCF-7 cells (22).","However, the sequence of the cloned protein (see FIG.","1) differed substantially from the p160 (Genbank U88153).","To differentiate between the two sequences, and to avoid confusion with the family of nuclear receptor coactivators that also are referred as p160s ((Voegel et al., 1998), (Hong et al., 1997), (Anzick et al., 1997), [Suen, 1998]), we named the protein that we cloned from MCF7 cells MNAR.","Sequence alignment of MNAR and p160 (FIG.","1A) indicated that MNAR does not contain nucleotides from 1075 to 1510 and 3125 to 3151 that are present in the p160 (Genbank U88153).","In addition, there were 10 single base pair gaps and one double base pair gap compared to the p160 sequence.","MNAR was also independently cloned from a human lymphoma Marathon cDNA library (Invitrogen).","Importantly, the MNAR cloned from MCF-7 cells was identical to the MNAR cloned using marathon cDNA library.","To obtain additional sequence information, we performed the 5′RACE with primers from marathon cDNA kit (Clonetech).","Using 5′RACE, stop codons were found in all three reading frames 5′ of the putative start codon of MNAR.","Conceptual translation of the MNAR clone resulted in an 1130 amino acid protein with a calculated molecular weight of 119.6 kDa.","This clone contained all peptide sequences initially identified by mass spectrometric analysis (FIG.","2B).","Sequence analysis has revealed multiple LXXLL motifs localized in the N-terminal portion of MNAR molecule.","We putatively call it nuclear receptor interaction domain (NRID).","Similar motifs in other transcription factors have been shown to interact with a hydrophobic groove on the surface of the ligand-binding domain of nuclear hormone receptors (Heery et al., 1997), (Torchia et al., 1997) (23).","Interesting features of the MNAR molecule are extended proline (PRD) and glutamic acid (ERD) rich domains localized in the C-terminal part of the MNAR molecule.","No homology to these domains has been found in the NCl database.","Example 2 MNAR Expression Analysis.","MNAR expression in HepG2, SaOS2, HOBs 02/02, COS-7, MCF-7.T47D, MDA231, Ishikawa, 1-10, LNCaPLN3, LNCaPFGC, JCA, TSUPRL cell lines was evaluated using Northern blotting analysis.","For this purpose we used an oligo-probe derived from the N-terminal portion of MNAR, which is common between MNAR and the deposited p160 sequence (Genbank U88153).","Expression of MNAR mRNA was detected in almost all cell lines (FIG.","3), as estimated by the presence of a 4 kb band (FIG.","3).","The level of the MNAR expression varied significantly among different cell lines.","It was high in liver (HepG2) and prostate (LNCaPLN3, LNCaPFGC, JCA, TSUPRL) cells, and low to undetectable in breast tumor cells that do not express ER (T47D, MDA231) (FIG.","3).","To evaluate whether p160 is expressed in MCF-7 cells we used Northern blotting analysis with oligo-probe to the region of p160 that is missing in MNAR sequence and is encoded by nucleotides 1480 to 1509 of deposited to Genbank p160 sequence.","Using this probe we failed to detect any message (data not shown).","This result indicates that the major form of this protein, expressed in MCF-7 cells, matches well with the MNAR, while the mRNA encoding the p160 is not expressed or expressed at very low levels.","MNAR distribution in different tissues was analyzed by Northern blotting.","Again, we used an oligo-probe derived from the N-terminal portion of MNAR, which is common between MNAR and the deposited p160 sequence (Genbank U88153).","Once again, expression of MNAR mRNA was detected in almost all tissues (FIG.","4), as estimated by the presence of a 4 kb band (FIG.","4).","The level of MNAR expression appears to vary among different tissues.","Example 3 MNAR Specifically Interacts with Estrogen Receptors With full-length flag-MNAR, we further evaluated whether MNAR interaction with estrogen receptors is affected by ER ligands.","Full-length flag-MNAR, overexpressed in SF9 cells using baculoviral expression system, was used in a pull-down experiment with in vitro transcribed/translated full-length unliganded ER α and β, or receptors liganded with E2, or 4-OH tamoxifen.","Formed complexes were isolated using anti-FLAG sepharose beads.","FIGS.","5A and 5B reveals that ER α and β interact with MNAR, and that this interaction is enhanced by E2 and inhibited by 4(OH) tamoxifen.","We also evaluated MNAR interaction with some other nuclear hormone receptors.","Full-length flag-MNAR was incubated with transcribed/translated androgen (AR) and glucocorticoid receptors (GR) with and without their corresponding hormone.","The data indicated that AR and GR ligand dependently interacted with MNAR (FIGS.","5B and 5C).","This interaction may explicate the nongenomic activity of these receptor ligands.","Detailed investigation of these and other nuclear hormone receptor's crosstalk with MNAR and the Src phosphorylation cascade is an important subject for future investigation.","Example 4 MNAR interaction with c-Src and Lck is enhanced by E2-liganded estrogen receptors.","Considering that a protein homologous to MNAR, p160, was initially identified via pull-down with the SH2 domain of p56lck (Lck), we next examined whether MNAR interacts with members of Src family tyrosine kinases and affects their interaction with ER.","To address this question, we have used in vitro transcribed/translated estrogen receptors ERα, ERβ and MNAR.","ER with and without 17β-estradiol (E2), alone or together with MNAR were incubated with commercially available, partially purified cSrc or Lck (Upstate Biotechnology).","Formed complexes were pulled-down with anti-cSrc or anti-Lck antibodies and protein A sepharose and then resolved on an SDS gel.","The data indicate that ER a interacts with cSrc and Lck (FIG.","6A, lanes 2 and 3, 6 and 7).","Surprisingly, this interaction was inhibited by E2.When ERa was incubated with both MNAR and cSrc (FIG.","6A, lanes 4 and 5) or Lck (FIG.","6A, lanes 8 and 9), significant stimulation of Src-ER-MNAR complex formation was detected in the presence of E2 (FIG.","6A, lanes 5 and 9) compared to unliganded ERa (FIG.","6A, lanes 4 and 8).","Since MNAR itself does not interact well with cSrc or Lck (FIG.","6A, lane 11), and interaction of cSrc and/or Lck with ERs is inhibited by E2 (FIG.","6A, lane 3,), we conclude that all three proteins strongly interact in the presence of E2 and that MNAR is the key to ER-Src interaction.","Identical data were obtained for ERβ (FIG.","6B).","Previous studies from Auricchio and colleagues suggested that ER directly binds to the SH2 domain of Src and that this binding is enhanced by E2.A potential explanation for the apparent inconsistency between our data and theirs may rely on the fact that to evaluate ER-Src interaction they used Src pull-downs from cell extracts.","MNAR was most likely present in these extracts and would have mediated ER-Src complex formation.","Under basal conditions, the catalytic domain of Src is constrained in an inactive state through intramolecular interactions.","Binding of the SH2 domain to the C-terminal phosphorylated tyrosine and SH3 domain to proline-rich region in Src linker domain locks the molecule in an inhibited conformation.","(FIG.","7).","Full catalytic activation requires release of these restraints.","The kinase activity of Src can be enhanced by binding of phosphotyrosine-containing sequences to the SH2 domain and binding of proline-rich sequences to the SH3 domain.","Considering that MNAR contains an extended proline-rich domain (PRD) and the ligand-binding domain of ER contains a tyrosine residue that can be phosphorylated, we hypothesized that these interactions may lead to Src activation (see FIG.","7).","Example 5 MNAR Stimulates ER-Driven Transactivation.","Activation of cSrc in response to EGF has been previously shown to promote ER phosphorylation and to promote phosphorylation of other transcription factors by MAP kinase, thereby stimulating ER-mediated transcription ([Kato, 1996]; [Bunone, 1996]).","To evaluate whether MNAR affects transcriptional activity of ER, COS7 cells were cotransfected with ERα (FIG.","6A) or ERβ (FIG.","6B) and MNAR.","Luciferase gene expression driven by 2×ERE-tk-reporter was evaluated in cells treated and untreated with 17β-estradiol.","Our data indicate that increased MNAR expression correlates with stimulation of ERα and ERβ transcriptional activity (FIGS.","6A and 6B).","Next we evaluated whether cSrc inhibitors affect MNAR-promoted ER activation.","HepG2 cells transfected with ERα and MNAR (FIG.","8) were treated for 24 hr with 17β-estradiol alone, with 17β-estradiol and selective inhibitor of Src—PP2 (Calbiochem) or with genistein.","As in the previous experiment (FIG.","8) MNAR stimulated E2 induced ERα activity.","At the same time Src inhibitors PP2 and genesteine inhibited MNAR stimulation (FIG.","8).","These data indicate that MNAR induced stimulation of ER activity is linked to the activation of a phosphorylation cascade.","Example 6 MNAR Overexpression affects Transcriptional Activity of ER To evaluate if MNAR overexpression affects transcriptional activity of ER, HepG2 cells were transiently cotransfected with plasmids for expression of ERα and MNAR.","Luciferase gene expression controlled by a 2×ERE-tk-reporter was evaluated in cells treated with E2, 4(OH)-tamoxifen, or ICI 182,780 all at 10 nM.","The results (FIG.","9A) indicate that increasing MNAR expression correlates well with stimulation of ERα transcriptional activity.","Similar stimulation of ER transcriptional activity was observed with overexpression of the ER-coactivator, SRC3.Our data also indicate that 4(OH)-tamoxifen and ICI 182,780 do not support MNAR stimulation of ER activity.","Similar results were observed with ERβ (data not shown).","To investigate if cSrc activation is responsible for the MNAR stimulation of ER activity, HepG2 cells transfected with ERα, MNAR and 2×ERE-tk-reporter plasmid (FIG.","9B) were treated with E2 alone, or E2 and either the cSrc inhibitor, PP2, or the MEK kinase inhibitor, PD98059.We found that both PP2 and PD98059 abrogate MNAR stimulation of ER activity.","These results indicate that MNAR promoted stimulation of ER transcriptional activity is linked to the activation of the MAP kinase phosphorylation cascade.","We hypothesize that ER phosphorylation by Erk, downstream from Src and MEK kinases, may be responsible for enhancement of ER transcriptional activity, as previously described.","In addition, it is possible the that activation of the Src/MAP kinase pathway leads to phosphorylation of some other transcription factors, that are important for ER transcriptional activity.","Importantly, these data also suggest that the so-called “nongenomic” action of nuclear hormone receptors through activation of the phosporylation cascade may regulate activity of transcription factors, and, by doing so, ultimately influence gene expression.","We speculate that this mechanism may institute the crosstalk between nuclear receptors that are able to interact with MNAR and other transcription factors whose activity is regulated by phosphorylation.","Example 7 MNAR Affects ER-Mediated Gene Expression We next used an antisense approach to assess MNAR's role in estrogen regulation of gene transcription.","Antisense oligonucleotides (SEQ ID NO:11) were developed (Sequitur Inc., Natick, Mass.)","that significantly inhibited MNAR expression in MCF-7 cells, as estimated by TaqMan (FIG.","9C) and Western Blotting (FIG.","9D) analyses.","MCF-7 cells were transfected with 100 nM antisense or reverse control oligomers.","After 24 hours, cells were stimulated with 10 nM E2, and 24 hours later cells were lysed and total RNA was isolated.","TaqMan analysis was performed using a primer/probe set designed to target MNAR, pS2 and Cathepsin D—two genes known to be regulated by estrogens in MCF-7 cells.","FIG.","9C presents the levels of MNAR, pS2 and Cathepsin D mRNA expression, normalized to GAPDH mRNA.","Interestingly expression of MNAR mRNA was substantially stimulated by E2 treatment.","Our data indicate that reduction in MNAR expression lead to dramatic reduction of E2-stimulated pS2 and Cathepsin D expression.","Importantly, the MNAR antisense oligonucleotides did not significantly affect the basal level of these genes expression.","We expect that ER phosphorylation is probably important for E2-mediated expression of pS2 and Cathepsin D. These data support our position that MNAR is essential for ER regulation of gene expression.","Example 8 MNAR and ER Interaction and Activation of Src Kinase in MCF-7 Cells We evaluated whether MNAR and ER interacted and activated Src kinase in MCF-7 cells.","MCF-7 cells transfected with a Flag-MNAR expression vector were treated with E2 at 10 nM for 5 minutes.","Flag-MNAR (FIG.","10, panel A), ERα (FIG.","10, panel C and D), and csrc (FIG.","10, panel B) were immunoprecipitated from the cell extracts using their corresponding antibodies.","Material precipitated using ERα antiserum was probed with rabbit polyclonal anti-MNAR antibody (FIG.","10, panel E).","E2-enhancement of ERα-MNAR interaction was clearly detected.","Considering that MCF7 cells not transfected with FLAG-MNAR were used for this experiment, this data confirm that endogenous MNAR and ERα interact in MCF7 cells.","Immunoprecipitates were incubated with [32P]γATP at 30° C. for 30 minutes.","Reaction was stopped by boiling in 2×SDS buffer and resolved on a SDS PMG.","Strong MNAR- and E2-dependent phosphorylation of several endogenous proteins was detected.","Especially evident was phosphorylation of an endogenous protein with an apparent molecular weight of approximately 34 kDa that coprecipitated with cSrc, ER, and MNAR (see FIG.","10).","Importantly phosphorylation of this protein was detected in material coprecipitated with anti-Src and anti-ER antibodies from cells untransfected with MNAR (FIG.","10, lanes 1 and 2).","The phosphorylation was stimulated in presence of E2 (FIG.","10, lane 2) and then dramatically augmented in cells transfected with MNAR (FIG.","10, lanes 3 and 4).","These data strongly support our position that MNAR, ER, and cSrc interact in MCF-7 cells, and that the MNAR-ER complex strongly stimulates cSrc kinase activity, promoting phosphorylation of some endogenous protein in MCF-7 cells.","An identical phosphorylation pattern was detected when enolase was used as Src substrate (FIG.","10, panel D).","Example 9 MNAR Affects E2 Induced Erk Kinase 1 and 2 Activation We next evaluated the level of Erk activation by E2 in MCF7 cells (see FIG.","11, lanes 3 and 4) cells overexpressing MNAR (FIG.","11, lanes 1 and 2) and cells in which MNAR expression was substantially attenuated by treatment with MNAR antisense oligonucleotide (FIG.","11, lanes 5 and 6).","Strong E2 induced stimulation of Erk 1 and 2 phosphorylation was clearly detected using antibodies that recognize phosphorylated Erk 1 and 2 (FIG.","11, panel A).","This activation was significantly enhanced in cells overexpressing MNAR (FIG.","11, panel A, lanes 1 and 2) and attenuated in cells treated with MNAR antisense oligonucleotides (FIG.","11, panel A, lanes 5 and 6).","Neither treatment with MNAR antisense oligonucleotides nor short treatment with E2 affected the level of total Erk 1 and 2 (FIG.","11, panel B).","Western blotting analysis indicated that MNAR level was strongly augmented in cells transfected with MNAR expression plasmid (FIG.","11, panel C, lanes 1 and 2) and attenuated in cells treated with MNAR antisense oligonucleotides (FIG.","11, panel C, lanes 5 and 6).","These data indicate that MNAR controls E2 induced stimulation of cSrc and Erk 1 and 2 kinases.","Example 10 MNAR Structural-Functional Organization MNAR sequence analysis revealed that the N-terminal portion of its molecule contains multiple LXXLL motifs found to be important for interaction with nuclear hormone receptors.","At the C-terminus, MNAR contains a region rich in both proline and glutamic acid.","To delineate the regions on MNAR responsible for interaction with estrogen receptors and c-src tyrosine kinase, a functional evaluation of MNAR deletion mutants was performed (FIG.","12).","A series of C-terminal truncations of MNAR was generated using specially designed oligonucleotides, by amplifying appropriate fragments and cloning them into appropriate expression vectors.","These mutants were tested for their abilities to enhance ER-stimulated transcription when transfected into HepG2 cells.","While MNAR mutants containing aa 1-469 (mutant 2, SEQ ID NO: 14), 1-278 (mutant 3, SEQ ID NO:15), and 1-189 (mutant 4, SEQ ID NO:16) are still able to augment ER transcriptional activity.","Mutants containing the C-terminal portion of MNAR molecule aa 595-1131 (mutant 9, SEQ ID NO: 17), 595-962 (mutant 10, SEQ ID NO: 18), and 595-887 (mutant 11, SEQ ID NO: 19)) were inactive.","These data suggest that the domain that is necessary and sufficient to stimulate ER transcriptional activity is localized to the N-terminal part of NRID between aa 1 and 189 (FIG.","13).","This region contains the LXXLL motifs—4th (aa154-159) (SEQ ID NO: 20), 5th (aa 176-181) (SEQ ID NO: 21), and 6th (aa 181-185) (SEQ ID NO: 22) (FIG.","12).","To evaluate which of these motifs is responsible for interacting with estrogen receptor, we generated mutants of MNAR with these three LXXLL motifs mutated individually or in combination.","While MNAR utilizes the LXXLL motifs to interact with estrogen receptors, MNAR uses PXXP motifs to interact with Src.","MNAR contains three of these PXXP motifs within the N-terminal portion.","From the deletion analysis, constructs of MNAR missing the first two PXXP motifs are not able to stimulate estrogen receptor activity in transfection assays (FIG.","13).","This suggests these first two PXXP motifs are sufficient for interaction with Src.","Mutation of these motifs revealed that the first PXXP motif (aa 55-58) (PXXP mutant 1, SEQ ID NO: 23) is necessary and sufficient to activate ER transcriptional activity, while the second PXXP motif (aa 64-67) (PXXP mutant 2 SEQ ID NO: 32) is dispensable (FIG.","14).","Additional mutants generated in this study were as follows: mutant 5 with aa 1-120 (SEQ ID NO: 24); mutant 6 with aa 1-79 (SEQ ID NO: 25); mutant 7 with aa 1-40 (SEQ ID NO: 26); mutant 8 with aa 190-469 (SEQ ID NO: 27); mutant 12 with aa 888-1131 (SEQ ID NO: 28); mutant 13 with aa 80-594 (SEQ ID NO: 29); and mutant 14 with aa 80-574 (SEQ ID NO: 30).","Each of these mutants, along with the other mutants listed above, were schematically diagrammed in FIG.","12 and were analyzed for their ability to enhance ER-stimulated transcription when transfected into HepG2 cells.","(FIG.","13) In conclusion, we demonstrate that interaction of estrogen receptors α and β with MNAR is enhanced by E2 but not by 4(OH) tamoxifen.","We also show that the MNAR-ER complex interacts with members of Src family of tyrosine kinases—p60src (Src) and p56lck (Lck) dramatically stimulating Src enzymatic activity.","We also show that MNAR, through activation of the Src phosphorylation cascade, affects ER transcriptional activity and ultimately ER-mediated gene expression.","Specifically, we demonstrate that short term treatment of MCF-7 cells with E2 leads to MNAR-Src-ER complex formation, activation of Src and phosphorylation of some endogenous proteins.","Enhanced MNAR expression in transiently transfected cells leads to stimulation of ER transcriptional activity.","Since specific inhibitors of Src and MEK kinases block the ability of MNAR to augment ER transcriptional activity, we conclude that this activation of ER is due to stimulation of the MAP kinase phosphorylation cascade promoted by activation of Src kinases.","Alternatively, depletion of MNAR from MCF-7 cells using antisense oligonucleotides inhibits ER-mediated gene expression, supporting its involvement in ER-regulated gene expression.","These data reveal that MNAR modulates ER crosstalk with Src family tyrosine kinases and by doing so mediates the non-genomic activity of estrogen receptors.","These studies demonstrate the importance of MNAR in interacting with nuclear receptors.","MNAR provides a means of developing compounds that selectively modulate genomic versus non-genomic activity of nuclear receptors.","All publications and patent applications cited in this specification are 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(1994).","Signal transduction by lymphocyte antigen receptors.","Cell 76, 263-74."]],"string":"[\n [\n \"Novel modulator of non-genomic activity of nuclear receptors (mnar) and uses thereof\",\n \"A novel protein modulator of non-genomic activity of nuclear receptors (MNAR), which interacts with estrogen receptors alpha (ERα), estrogen receptor beta (ERβ), and other nuclear receptors, was discovered and characterized.\",\n \"An MNAR-ER complex, in vitro and in vivo, relates to nucleic acids, antisense, recombinant expression of nucleic acids, as well as host cells, compositions and assays that utilize the nucleic acids and protein of this invention.\"\n ],\n [\n \"1.An isolated nucleic acid sequence comprising a polynucleotide selected from the group consisting of: (a) a polynucleotide encoding a novel protein modulator of non-genomic activity of nuclear receptors (MNAR) comprising the amino acid sequence of SEQ ID NO:2 or of SEQ ID NO:13; (b) a polynucleotide that hybridizes under highly stringent conditions with (i) a region of the nucleotide sequence of SEQ ID NO: 1 or of SEQ ID NO:12, (ii) a subsequence of at least 100 nucleotides of the nucleotide sequence of SEQ ID NO: 1 or of SEQ ID NO:12, (iii) or a complementary strand of (i) or (ii); (c) a polynucleotide comprising a sequence with at least 85% identity to a polynucleotide coding sequence of SEQ ID NO: 1 or of SEQ ID NO: 12; (d) a variant of the polynucleotide comprising a polynucleotide coding sequence of SEQ ID NO:1 or of SEQ ID NO:12; and (e) a polynucleotide encoding a polypeptide fragment comprising the amino acids of SEQ ID NOS: 20 or 23 (f) a complement or antisense form of the polynucleotide of (a), (b), (c), (d) or (e); wherein the polynucleotide does not encode a p160 protein: 2.The isolated nucleic acid sequence of claim 1, wherein the polynucleotide sequence is an allelic variant.\",\n \"3.A polypeptide comprising an amino acid sequence selected from the group consisting of: (a) an amino acid sequence for a protein modulator of non-genomic activity of nuclear receptors (MNAR) comprising the amino acid sequence of SEQ ID NO: 2 or of SEQ ID NO:13; (b) an amino acid sequence for an MNAR fragment comprising the amino acid sequence of SEQ ID NOS:14, 15 or 16; (c) a variant of (a) or (b), and (d) a fragment of (a), (b), or (c).\",\n \"4.The polypeptide of claim 3 comprising one or more amino acid sequences which are more than 90% identical to the amino acid sequence of SEQ ID NO:14.5.The polypeptide of claim 3 comprising one or more amino acid sequences which are more than 95% identical to the mutant sequence of SEQ ID NO:14.6.The polypeptide of claim 3, wherein said polypeptide comprises the amino acid sequence of SEQ ID NOS 2, 13, 14, 15 16 20 or 22.7.A fusion polypeptide comprising a polypeptide, or polypeptide fragment of MNAR.\",\n \"8.The polypeptide according to claim 3, wherein said polypeptide stimulates the activity of a tyrosine kinase in the presence of one or more nuclear receptors.\",\n \"9.The polypeptide of claim 8, wherein said nuclear receptor is a steroid receptor.\",\n \"10.The polypeptide of claim 8, wherein said nuclear receptor is an estrogen receptor.\",\n \"11.The polypeptide of claim 8, wherein said estrogen receptor is estrogen receptor alpha.\",\n \"12.An expression vector comprising the isolated nucleic acid sequence of claim 1.13.The expression vector of claim 12 comprising the polynucleotide sequence of SED ID NO: 1 or of SEQ ID NO:12.14.A host cell transformed with the vector of claim 12.15.The host cell of claim 14, wherein the host cell is a mammalian host cell.\",\n \"16.An isolated antibody to the polypeptide of claim 3.17.The antibody of claim 16, wherein said antibody is a monoclonal antibody.\",\n \"18.The antibody of claim 16, wherein said antibody is a polyclonal antibody.\",\n \"19.A method for modulating the transcriptional activity of a nuclear receptor, comprising providing to the loci of a nuclear receptor a polypeptide according to claim 3.20.A method for modulating the transcriptional activity of a nuclear receptor, comprising: (a) transforming a host cell with the recombinant vector having the nucleic acid sequence for MNAR according to claim 1; and (b) culturing said transformed host cell to express the nucleic acid in the presence of a nuclear receptor.\",\n \"21.A method of identifying compounds exhibiting non-genomic activity versus genomic activity comprising administering a test compound to a cell comprising an MNAR-nuclear receptor complex and measuring the non-genomic or genomic activity resulting from the test compound.\",\n \"22.The method of claim 21, wherein at least one ligand of ER is present in the cell, or a kinase is present, or both are present.\",\n \"23.A method of screening for a compound that modulates the activity of MNAR on a nuclear receptor, comprising the steps of (a) contacting a test compound with the polypeptide of claim 3; and (b) determining whether said test compound specifically binds said polypeptide.\",\n \"24.A method of screening for a compound that modulates the activity of MNAR on a nuclear receptor, said method comprising the steps of (a) adding a test compound to a cell comprising the polypeptide of claim 3 and a nuclear receptor; and (b) comparing the MNAR activity before and after said adding step.\",\n \"25.The method of claim 24, said method comprising adding a test compound to a control comprising a mutant cell lacking MNAR activity or with significantly reduced MNAR activity.\",\n \"26.A method for detecting the non-genomic activity of a compound comprising administering a compound to a cell in the presence of MNAR-nuclear receptor and in the absence of MNAR and comparing the level of non-genomic activity.\",\n \"27.A method for detecting the genomic activity of a compound comprising administering a compound to a cell comprising a MNAR-nuclear receptor complex and comparing the level of genomic activity in the absence of MNAR.\",\n \"28.A method for detecting the genomic activity of a compound according to claim 51 wherein the MNAR-ER cell overexpresses MNAR.\",\n \"29.A method for detecting the genomic activity of a compound according to claim 51 wherein the MNAR-ER cell overexpresses ER.\",\n \"30.A method for detecting the genomic activity of a compound according to claim 27 wherein the ER is operatively associated with a reporter.\",\n \"31.A method of identifying compounds having selective genomic versus non-genomic activity said method comprising the steps of (a) adding a test compound to a cell comprising a MNAR-nuclear receptor complex; and (b) comparing the genomic versus non-genomic activity before and after said adding the test compound.\",\n \"32.The method of claim 59, wherein selective genomic activity is measured as a two-fold increase after addition of said test compound to a cell in the presence of MNAR-nuclear receptor when compared to genomic activity with test compound in the absence of MNAR and wherein no change is observed in non-genomic activity after addition of said test compound.\",\n \"33.The method of claim 31, wherein non-genomic activity is increased two-fold after addition of said test compound to a cell in the presence of MNAR-nuclear receptor complex when compared to non-genomic activity with test compound in the absence of MNAR and wherein no change is observed in genomic activity after addition of said test compound.\",\n \"34.A method of identifying compounds that modulate the non-genomic activity of a nuclear receptor, comprising determining an increase or decrease in the transcriptional activity of a nuclear receptor in the presence of MNAR.\",\n \"35.A method of determining whether a desired phenotype of cell is effected by non-genomic activity or genomic activity of a nuclear receptor comprising determining the effect of a selected phenotype of cell is increased or decreased when the cell is exposed to additional amounts of MNAR; and then detecting the presence or absence of a desired phenotype, wherein the absence of a desired activity or phenotype, in the presence of additional MNAR indicates increased non-genomic activity of a nuclear receptor negatively affects the desired phenoype (i.e.\",\n \"the non-genomic activity is not beneficial to said cell) and wherein the increase of a desired phenotype indicates that increased non-genomic activity of a nuclear receptor positively affects said cell.\",\n \"36.A method according to claims 2, wherein the cell employed is cell that is positively affected by non-genomic activity of the nuclear receptor.\",\n \"37.A method according to claims 21, wherein the cell employed is cell that is negatively affected by non-genomic activity of the nuclear receptor.\",\n \"38.A method of identifying compounds that modulate the non-genomic activity of a nuclear receptor, comprising determining an increase or decrease in the transcriptional activity of said nuclear receptor and determining whether a kinase, affected by the presence of MNAR and said nuclear receptor, has an increase or decrease in enzymatic activity.\",\n \"39.The method of claim 19, wherein the nuclear receptor is a steroid receptor.\",\n \"40.The method of claim 19, wherein the nuclear receptor is a non-steroid receptor.\",\n \"41.The method of claim 19, wherein the nuclear receptor is an orphan receptor.\",\n \"42.The method of claim 19, wherein the steroid receptor is an estrogen receptor.\",\n \"43.A non-human transgenic mammal, comprising one or more cells which comprise at least one non-functional endogenous MNAR polynucleotide sequence.\",\n \"44.The non-human transgenic mammal of claim 43, wherein said mammal is selected from the group consisting of a mouse, a rat, a hamster, a guinea pig, a rabbit, a cow, and a sheep.\",\n \"45.The non-human transgenic mammal of claim 43, comprising at least one non-functional endogenous nuclear receptor.\",\n \"46.The non-human transgenic mammal of claim 43, wherein the non-functional endogenous receptor is a steroid receptor.\",\n \"47.The non-human transgenic mammal of claim 43, wherein the non-functional endogenous receptor is a non-steroid receptor.\",\n \"48.The non-human transgenic mammal of claim 43, wherein the non-functional endogenous receptor is an orphan receptor.\",\n \"49.The non-human transgenic mammal of claim 43, wherein said the non-functional endogenous receptor is an estrogen receptor.\",\n \"50.The non-human transgenic mammal of claim 49, wherein endogenous forms of ERα and ERβ are non-functional.\",\n \"51.The non-human transgenic mammal of claim 43, comprising an interruption of the MNAR coding sequence.\",\n \"52.The non-human transgenic mammal of claim 49, comprising an interruption of the MNAR and ER coding sequences.\",\n \"53.The non-human transgenic mammal of claim 43, comprising a nonsense MNAR mutation.\",\n \"54.The non-human transgenic mammal of claim 49, comprising a nonsense mutations of MNAR and ER.\",\n \"55.The non-human transgenic mammal of claim 43, comprising a deletion of MNAR coding sequences.\",\n \"56.The non-human transgenic mammal of claim 43, comprising an alteration in the regulatory region of the MNAR gene making MNAR non-functional.\",\n \"57.The non-human transgenic mammal of claim 56, comprising substitution of an inducible/repressable promoter for the endogenous MNAR promoter.\",\n \"58.The non-human transgenic mammal of claim 57, comprising at least one inducible/repressable promoter replacing the endogenous MNAR promoter and the estrogen receptor.\",\n \"59.The non-human transgenic mammal of claim 43, wherein cells of said mammal further comprise an exogenous selectable marker gene under the control of a promoter active in at least one cell type of said mammal.\",\n \"60.A method of screening for a compound that disrupts the function of MNAR, comprising using a protein-protein interaction assay to identify compounds that interact with MNAR.\",\n \"61.A method of screening for a compound that disrupts the function of MNAR, comprising using a two-hybrid assay to identify proteins that interact with MNAR.\",\n \"62.The method of claim 61 wherein the two-hybrid is conducted in yeast or mammalian cells.\",\n \"63.A process for producing a polypeptide of claim 3 comprising: (a) culturing a host cell transformed with a nucleic acid sequence for MNAR according to claim 1 under suitable culture medium under conditions for growth; and (b) isolating the polypeptide from the culture medium.\",\n \"(c) 64.A polypeptide produced from the process of claim 64.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>Nuclear hormone receptors are a superfamily of ligand-inducible transcription factors, which, as a class, are involved in ligand-dependent transcriptional control of gene expression.\",\n \"Binding of a specific ligand, inducing conformational changes in the receptor molecule, affects receptor interaction with other transcription factors, and, ultimately, formation of the preinitiation complex.\",\n \"This process regulates the rate of gene transcription.\",\n \"(D. J. Mangelsdorf, et al., Cell 83: 835-9, 1995).\",\n \"The nuclear hormone receptor superfamily includes steroid hormone receptors, non-steroid hormone receptors, and orphan receptors.\",\n \"The receptors for glucocorticoids (GR), mineralcorticoids (MR), progestins (PR), androgens (AR), and estrogens (ER) are examples of classical steroid receptors.\",\n \"In addition to steroid hormone receptors, this nuclear hormone receptor superfamily consists of receptors for non-steroid hormones, such as vitamin D, thyroid hormones, and retinoids.\",\n \"Moreover, a range of nuclear receptor-like sequences have been identified which encode so called “orphan” receptors.\",\n \"These orphan receptors are structurally related to and, therefore, classified as nuclear hormone receptors, although no putative ligands have been identified yet.\",\n \"(Cell, 83: 851-857, 1995).\",\n \"The superfamily of nuclear hormone receptors is characterized structurally and functionally by a modular structure in comprising six distinct structural and functional domains, A to F. More specifically these receptors have a variable N-terminal region (domain A/B); followed by a centrally located, highly conserved DNA-binding domain (hereinafter referred to as DBD; domain C); a variable hinge region (domain D); a conserved ligand-binding domain (hereinafter referred to as LBD; domain E); and a variable C-terminal region (domain F).\",\n \"Cell, supra.\",\n \"The N-terminal region, which is highly variable in size and sequence, is poorly conserved among the different members of the superfamily.\",\n \"This part of the receptor is involved in the modulation of transcription activation.\",\n \"The DBD consists of approximately 66 to 70 amino acids and is responsible for DNA-binding activity: This domain targets the receptor to specific DNA sequences, called hormone responsive elements (hereinafter referred to as HRE), within the transcription control unit of specific target genes on the chromatin.\",\n \"Steroid receptors such as GR, MR, PR, and AR recognize similar HRE DNA sequences, while the ER recognizes a different DNA HRE sequence.\",\n \"After binding to DNA, the steroid receptor is thought to interact with components of the basal transcriptional machinery and with sequence-specific transcription factors, thereby modulating the expression of specific target genes.\",\n \"The LBD is located in the C-terminal part of the receptor and is primarily responsible for ligand binding activity.\",\n \"In this way, the LBD is essential for recognition and binding of the hormone ligand and, in addition, possesses a transcription activation function, thereby determining the specificity and selectivity of the hormone response of the receptor.\",\n \"Although moderately conserved in structure, the LBD's are known to vary considerably in homology among the individual members of the nuclear hormone receptor superfamily.\",\n \"When a hormone ligand for a nuclear receptor enters the cell and is recognized by the LBD, it will bind to the specific receptor protein, thereby initiating an allosteric alteration of the receptor protein.\",\n \"(Cell, supra).\",\n \"As a result of this alteration the ligand/receptor complex switches to a transcriptionally active state and, as such, is able to bind through the presence of the DBD with high affinity to the corresponding HRE on the chromatin DNA.\",\n \"In this way, the ligand/receptor complex modulates expression of the specific target genes.\",\n \"The diversity achieved by this family of receptors results from their ability to respond to different ligands.\",\n \"In addition to modulation of genomic activity, hormone receptor complexes can have important and varied non-genomic effects.\",\n \"This non-genomic activity is characterized by fast and transient increase in intracellular second messengers.\",\n \"In turn, these second messengers are involved in a number of different pathways and cascades that affect cellular functions, such as proliferation and differentiation.\",\n \"More generally, steroid hormone receptors are connected with embryonic development, adult homeostasis, and organ physiology.\",\n \"Various diseases and abnormalities are ascribed to a disturbance in steroid hormone action.\",\n \"Since steroid receptors exercise their influence as hormone-activated transcription modulators and as hormone-activated stimulators of non-genomic activity, further investigation into various approaches to modifying, interacting with, or modulating these receptors is an area of immense significance.\",\n \"For example, mutations and defects in these receptors, as well as overstimulation or blocking of these receptors, may provide better insight into the underlying mechanism of the hormone signal transduction pathway, thereby leading to increased efficacy in treatment of a wide variety of steroid receptor-linked diseases and abnormalities.\",\n \"Estrogen Receptors Estrogen (E2) exerts numerous biological effects in different tissues through an interaction with the ER.\",\n \"Amino acid sequence analyses, transient transfection studies, and mutational dissections of ER indicate that ER has the classical modular structure described above.\",\n \"The N-terminal A/B domain of the ER contains a transactivation function, referred to as transcriptional activation function 1 (TAF-1).\",\n \"The DBD contains two zinc fingers and is responsible for DNA recognition.\",\n \"The LBD and a second transactivation function, referred to as TAF-2, is located at the C-terminal of ER.\",\n \"Upon binding to hormone, the ER undergoes an activation and transformation step.\",\n \"The activated ER interacts with specific estrogen response elements (EREs) that are located in the promoter region of estrogen-regulated genes and that influence its target gene transcription.\",\n \"Over the past decade, numerous studies have provided a basic understanding of both the effects of ligand (agonist/antagonist) on the ER and the relationship between the structure and function of the ER.\",\n \"Nevertheless, little is known regarding the mechanisms for the non-genomic activity of ER.\",\n \"ERβ appears to be distinct from the more commonly known estrogen receptor, referred to as ERα.\",\n \"Collectively, ERα and ERβ are referred to herein as ER.\",\n \"The DBD of ERβ is 90% identical to that of ERα.\",\n \"However, the overall homology between the ligand binding domain (LBD) of ERα and ERβ is less than 55%.\",\n \"Like ERα, ERβ can stimulate transcription from an ERE in a ligand-dependent manner.\",\n \"It has been established that estrogens induce fast and transient increases in the levels of intracellular second messengers, including calcium and cAMP, and that estrogens induce activation of mitogen-activated protein kinase (MAPK) and phospholipase C (Collins and Webb, 1999).\",\n \"In fact, numerous studies have demonstrated that estrogens induce rapid and transient activation of the Src/Ras/MAP kinase phosphorylation pathway.\",\n \"Activation of this pathway triggers vital cellular functions including cell proliferation and differentiation.\",\n \"The time course of these acute events parallels that elicited by peptide hormones, thus supporting the hypothesis that these events do not involve the ‘classical’ genomic action of estrogens.\",\n \"Recent data also suggests a direct link between the estrogen receptor and the mitogen-activated protein (MAP) kinase-signaling cascade.\",\n \"MAP kinases are a family of serine-threonine kinases that are phosphorylated and activated in response to a variety of signals.\",\n \"These enzymes transduce extracellular signals from multiple membrane receptors to intracellular targets, including transcription factors, cytoskeletal proteins and enzymes.\",\n \"The MAP kinase family includes the extracellular-signal related kinases (ERKs), p38 and cJun N-terminal kinases which signal through a pathway involving sequential activation of Ras, Raf and mitogen-activated protein kinase (MEK).\",\n \"(S. M. Thomas, J. S. Brugge, Annual Review of Cell & Developmental Biology 13, 513-609, 1997).\",\n \"In pulmonary endothelial cells, neuronal cells, osteoblasts and osteoclasts, 17β-estradiol (E2) has been reported to rapidly activate the MAPK pathway.\",\n \"More specifically, in pulmonary endothelial cells, 17β-estradiol (E2) has been reported to rapidly stimulate nitric oxide (NO) production, thus explaining the ability of E2 to induce acute dilation of blood vessels.\",\n \"Chen et al.\",\n \"(Chen et al., 1999) have recently reported that E2 induces rapid activation of endothelial nitric oxide synthase (eNOS) in isolated pulmonary endothelial cells.\",\n \"Estrogen activation of eNOS was shown to occur through the rapid activation of the MAP kinase pathway.\",\n \"Complimentary studies have also shown that E2 induces calcium-dependent translocation of eNOS from the plasma membrane to intracellular sites close to the nucleus—an action that is rapid (within 5 minutes), receptor-mediated but nongenomic (Goetz et al., 1999).\",\n \"In addition, ligand-dependent ERα interaction with the p85α regulatory subunit of phosphatidylinositol-3-OH kinase (PI(3)K) was recently implicated in mediation of cardiovascular protective effects of estrogen (Simoncini T. and J. K., 2000).\",\n \"Stimulation with estrogen increases ERα associated PI(3)K activity, thereby leading to the activation of protein kinase B/Akt and endothelial nitric oxide synthase (eNOS).\",\n \"In neuronal cells, rapid activation of the MAP kinase signaling pathway by E2 results in neuroprotection in primary cortical neurons after glutamate excitotoxicity (Singer et al., 1999).\",\n \"These neuroprotective effects of estrogens, which were reported to occur within 5 minutes after exposure, were mediated through the transient activation of c-Src-tyrosine kinases and tyrosine phosphorylation of p21 (ras)-guanine nucleotide activating protein in an ER-dependent manner.\",\n \"Similarly, in osteoblasts E2 induces MAP kinase phosphorylation and activation within 5 minutes (Endoh et al., 1997).\",\n \"MAP kinase activation by estrogens may, therefore, regulate cell proliferation and differentiation in these cells leading to increased bone formation.\",\n \"Several studies also indicate that estrogens activate the MAP kinase pathway in osteoclasts through activation of Src kinase (Oursler, 1998).\",\n \"It has been also shown that MAP kinase activation, could be involved in the regulation of enzymes needed for bone resorption (Kristen D. Brubaker, 1999).\",\n \"In the human mammary cancer derived cell lines, MCF-7 and T47D, as well as in the human colon cancer derived cell line, Caco-2, E2 activates the signal transducing Src/Ras/Erk pathway (Migliaccio et al., 1993), (Migliaccio et al., 1996), (Migliaccio et al., 1998), (Migliaccio et al., 2000).\",\n \"This activation is mediated by direct ER-Src interaction.\",\n \"Interestingly, progesterone also activates the same pathway in T47D cells (Migliaccio et al., 1998).\",\n \"The Src/Ras/ERk signaling pathway is a well-known target of growth factors.\",\n \"Importantly, activation of this pathway requires direct interaction of ER with Src.\",\n \"Activation of this pathway triggers different cellular responses such as proliferation or differentiation [Cantley, 1991]; [Marshall, 1996]; [Downward, 1997].\",\n \"Its activation by estrogen receptor ligands, such as steroid hormones, explicates their involvement in the cell cycle control.\",\n \"These data support the view that non-transcriptional/nongenomic activity of ER may be responsible for stimulation of cell growth.\",\n \"However, the details of the molecular mechanism of this process are largely unknown.\",\n \"Src activation has been previously observed in a large number of human breast and colon carcinomas ([Rosen, 1986]; [Ottenhoff-Kalff, 1992]).\",\n \"Activation of Src induces mammary tumors in transgenic mice ([Guy, 1994]).\",\n \"Constitutively active Ras mutants have been found in 25-30% of all cancers, including breast cancer ([Kasid, 1987]).\",\n \"Recent studies have suggested the existence of a plasma membrane estrogen receptor unrelated to the classical ER.\",\n \"However, cloning or isolation of this membrane ER has not been accomplished, while others have suggested that a subpopulation of the classical ER is associated with the cell membrane and is responsible for the rapid effects of estrogens.\",\n \"Transcriptional activity of nuclear receptors itself is a target for tight regulation by multiple signaling pathways, including those stimulated by the neurotransmitter dopamine ([Power, 1991]; [Smith, 1993]), growth factors such as epidermal growth factor (EGF), transforming growth factor-α (TGF-a) and insulin-like growth factor I (IGF-I) ([Ignar-Trowbridge, 1996], [Aronica, 1993]; [Bunone, 1996]), and by activators of protein kinase C ([Aronica, 1994]).\",\n \"The molecular mechanisms for such cross coupling are thought to be mediated, at least in part, by receptor phosphorylation.\",\n \"The ER has been demonstrated to be phosphorylated by MAP kinase in response to EGF, thereby resulting in stimulation of AF1 ([Kato, 1996]; [Bunone, 1996]).\",\n \"Other studies have demonstrated that the ER ([Auricchio, 1987]; [Arnold, 1995]; [Pietras, 1995]), Thyroid hormone receptor (TRβ) ([Lin, 1992]), RARγ ([Rochette-Egly, 1992]), glucocorticoid receptor ([Rao, 1987]) and the orphan receptor HNF-4 ([Ktistaki, 1995]) are also targets for tyrosine phosphorylation.\",\n \"A specific tyrosine phosphorylation site has been identified within the AF2 domain of the human ER (hER) at amino acid 537 (Y537) ([Castoria, 1993]; [Arnold, 1995]).\",\n \"This tyrosine is located immediately N-terminal of the AF2 sequence and is conserved in all known ER sequences from diverse species including the ERβ ([Kuiper, 1996]; [Mosselman, 1996]).\",\n \"Although the molecular mechanisms of non-genomic and genomic activities of E2 are not well understood, the present invention should allow us to create a new generation of therapeutics with tissue and gene selective activity.\",\n \"To better understand the tissue selective action of some ER ligands, we evaluated the expression and activity of ER-interacting proteins in different cell lines.\",\n \"Using affinity purification and mass spectrometer based microsequencing, we discovered a novel ER interacting protein, designated as MNAR (modulator of non-genomic activity of nuclear receptors).\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>The present invention provides for a novel protein modulator of non-genomic activity of nuclear receptors (MNAR) and nucleic acids that encode such proteins.\",\n \"The MNAR protein can interact with nuclear hormone receptors, such as estrogen receptors.\",\n \"In general, the protein or polypeptide of MNAR refers to the amino acid sequence of an MNAR.\",\n \"In particularly preferred embodiments, MNAR protein interacts with ERα and ERβ.\",\n \"This interaction is ligand dependent.\",\n \"MNAR will also form a complex with ER and kinases, such as members of the Src family.\",\n \"Src and MAP kinase activation leads to enhancement of ER transcriptional activity.\",\n \"The present invention provides (a) a novel nucleic acid comprising a polynucleotide encoding an MNAR having a putative amino acid sequence of SEQ ID No: 2 (see FIG.\",\n \"12 ).\",\n \"A nucleotide sequence of human MNAR is shown in SEQ ID No: 1 (See FIG.\",\n \"15 ).\",\n \"An amino acid sequence of human MNAR is shown in SEQ ID NO: 13 (See FIG.\",\n \"16 ).\",\n \"A nucleotide sequence of murine MNAR is shown in SEQ ID NO: 12 (See FIG.\",\n \"17 ).\",\n \"An amino acid sequence of murine MNAR is shown in SEQ ID NO: 13 (See FIG.\",\n \"18 ).\",\n \"This invention provides novel isolated nucleic acid comprising a polynucleotide selected from the group consisting of: (a) a polynucleotide encoding a novel protein modulator of non-genomic activity of nuclear receptors (MNAR) comprising the amino acid sequence of SEQ ID NO:2 or of SEQ ID NO:13; (b) a polynucleotide that hybridizes under highly stringent conditions with (i) a region of the nucleotide sequence of SEQ ID NO: 1 or of SEQ ID NO:12, (ii) a subsequence of at least 100 nucleotides of the nucleotide sequence of SEQ ID NO: 1 or of SEQ ID NO:12, (iii) or a complementary strand of (i) or (ii); (c) a polynucleotide comprising a sequence with at least 85% identity to a polynucleotide coding sequence of SEQ ID NO: 1 or of SEQ ID NO: 12; (d) a variant of the polynucleotide comprising a polynucleotide coding sequence of SEQ ID NO:1 or of SEQ ID NO:12; and (e) a polynucleotide encoding a polypeptide fragment comprising the amino acids of SEQ ID NOS 20 or 23 (f) a complement or antisense form of the polynucleotide of (a), (b), (c), (d) or (e); wherein the polynucleotide does not encode a p160 protein ( FIG.\",\n \"1 ).\",\n \"In preferred embodiments the isolated nucleic acid comprises an allelic variant of the polynucleotide.\",\n \"In certain embodiments, the nucleic acid is operably linked to an expression control sequence (e.g.\",\n \"heterologous or homologous control sequences).\",\n \"This invention provides for a host cell, including bacteria, yeast, insect, and mammalian cells transformed with such isolated nucleic acids.\",\n \"This invention also relates to a polypeptides comprising an amino acid sequence selected from the group consisting of: (a) an amino acid sequence for a protein modulator of non-genomic activity of nuclear receptors (MNAR) comprising the amino acid sequence of SEQ ID NO: 2 or of SEQ ID NO:13; (b) an amino acid sequence for an MNAR fragment comprising the amino acid sequence of SEQ ID NOS:14, 15 or 16; (c) a variant of (a) or (b), and (d) a fragment of (a), (b), or (c).\",\n \"In alternative embodiments, these polypeptides comprise an amino acid sequences which is more than 90%, or 95% identical to the amino acid sequence of SEQ ID NO:13 or 14.In particular embodiments, the polypeptides of the invention comprise the amino acid sequence of SEQ ID NOS 2, 13, 14, 15 16 20 or 22.In additional embodiments the polypeptides of this invention can stimulates the activity of a tyrosine kinase in the presence of one or more nuclear receptors.\",\n \"The nuclear receptors can be selected from steroid and non-steroid nuclear receptors as well as orphan nuclear receptors.\",\n \"In certain embodiments, the nuclear receptor is an estrogen receptor.\",\n \"Alternative embodiments involve monoclonal and polyclonal antibodies which correspond to MNAR polypeptides as well as fragments thereof.\",\n \"In view of the modulation by MNAR of nuclear receptors, many embodiments of this invention relate to modulating the transcriptional activity of a nuclear receptor, comprising providing to the loci of a nuclear receptor an MNAR polypeptide.\",\n \"For example, one can modulate the transcriptional activity of a nuclear receptor, by (a) transforming a host cell with the recombinant vector having the nucleic acid sequence for MNAR according to claim 1 ; and (b) culturing said transformed host cell to express the nucleic acid in the presence of a nuclear receptor.\",\n \"This invention also presents numerous approaches for identifying compounds that affect non-genomic activity or genomic activity of nuclear receptors.\",\n \"In one embodiment, a method of identifying compounds exhibiting non-genomic activity versus genomic activity compries administering a test compound to a cell comprising an MNAR-nuclear receptor complex and measuring the non-genomic or genomic activity resulting from the test compound.\",\n \"In these methods, preferably, at least one ligand of a nuclear receptor is present in the cell, or a kinase is present, or both are present.\",\n \"Other methods are directed to the screening for a compound that modulates the activity of MNAR on a nuclear receptor, comprising the steps of (a) contacting a test compound with an MNAR polypeptide; and (b) determining whether said test compound specifically binds said polypeptide.\",\n \"In addition, one can conduct a method of screening for a compound that modulates the activity of MNAR on a nuclear receptor, said method comprising the steps of (a) adding a test compound to a cell comprising the polypeptide of and a nuclear receptor; and (b) comparing the MNAR activity before and after said adding step.\",\n \"Further approaches to this method involve adding a test compound to a control comprising a mutant cell lacking MNAR activity or with significantly reduced MNAR activity.\",\n \"In each of these methods one can use a control to assess the non-genomic activity (or genomic activity) of a compound by administering a compound to a cell in the presence of MNAR and a nuclear receptor, and then repeating the experiment in the absence of MNAR and comparing the level of non-genomic activity.\",\n \"Preferred methods employ a cell overexpresses MNAR a nuclear receptor or both.\",\n \"The genomic activity of a compound is detected by having the nuclear receptor operatively associated with a reporter.\",\n \"Certain embodiments of the present invention also relate to a method of identifying compounds having selective genomic versus non-genomic activity said method comprising the steps of (a) adding a test compound to a cell comprising a MNAR-nuclear receptor complex; and (b) comparing the genomic versus non-genomic activity before and after said adding the test compound.\",\n \"A selective genomic activity can be measured by conventional means.\",\n \"Preferably, the increase or positive effect of non-genomic activity, as measured, is a two-fold increase after addition of said test compound to a cell in the presence of MNAR-nuclear receptor when compared to genomic activity with test compound in the absence of MNAR and wherein no change is observed in non-genomic activity after addition of said test compound.\",\n \"On the other hand, when selecting for test compounds for non-genomic activity, there should be at least about a two-fold increase after addition of said test compound to a cell in the presence of MNAR-nuclear receptor complex when compared to non-genomic activity with test compound in the absence of MNAR and wherein no change is observed in genomic activity after addition of said test compound.\",\n \"One can measure the effect by determining an increase or decrease in the transcriptional activity of a nuclear receptor in the presence of MNAR.\",\n \"In a variety of cells, healthy or diseased, it can be valuable to understand whether a beneficial or healthy state for a cell results from non-genomic or genomic activity.\",\n \"The present invention permits such determinations by comparing a desired phenotype of a cell, in the presence and absence of MNAR.\",\n \"The present invention also relates to non-human transgenic mammals, wherein one or more cells comprise at least one non-functional endogenous MNAR polynucleotide sequence, at least one non-functional endogenous nuclear receptor or both.\",\n \"In preferred embodiments, the non-human transgenic mammal do not have functional endogenous forms of ERa and ERA.\",\n \"Alternatively, transgenic mammal comprises substitution of an inducible/repressable promoter for the endogenous MNAR promoter.\",\n \"The non-human transgenic mammal of claim may further comprise an exogenous selectable marker gene under the control of a promoter active in at least one cell type of said mammal.\",\n \"This invention also relates to a method of screening for a compound that disrupts the function of MNAR, comprising using a protein-protein interaction assay to identify compounds that interact with MNAR.\",\n \"Once can employ any of the well-known two-hybrid assays or other conventional assays to study protein-protein interactions and the disruption thereof by test compounds, which can be a natural product isolate, small molecule chemical, peptide, etc.\",\n \"MNAR protein and MNAR encoding nucleic acids can be obtained from mammalian cells and tissues (e.g.\",\n \"human and murine cells and tissues).\",\n \"Recombinant MNAR and anti-MNAR antibodies find use in drug screening, diagnostics, and therapeutics.\",\n \"In particular, the MNAR provides valuable reagents in developing specific biochemical assays for screening compounds for ER ligands that agonize or antagonize the enzymatic activity of the Src family of tyrosine kinases, which are involved in regulating gene expression.\",\n \"Additionally, the MNAR provides valuable reagents in developing specific biochemical assays for screening compounds for a variety of nuclear receptor ligands.\",\n \"Accordingly, one can analyze the effects on any pathway involving a nuclear receptor, such as ER, and/or a kinase.\",\n \"For example, one can use MNAR to set up assays to determine the effect of ER ligands or test compounds on (I) cell cycle stimulation in cancer cells; (II) cell proliferation, cell differentiation and cell conservation for bone development or modulation; (III) neuroprotective activity of ER ligands/test compounds in nerve cells; and (IV) pulmonary characteristics of endothelial cells.\"\n ],\n [\n \"This application claims the benefit of U.S. application Ser.\",\n \"No.\",\n \"60/281,155, filed Apr.\",\n \"3, 2001.This application is a continuation in part of the aforementioned patent application, the content of which is incorporated by reference.\",\n \"FIELD OF THE INVENTION The present invention relates to the fields of biochemistry and internal medicine and to a novel modulator of non-genomic activity of nuclear receptors (MNAR), which interacts with both estrogen alpha (ERα) and estrogen beta (ERβ).\",\n \"More particularly, this invention concerns novel compositions, methods, and assays for modulating the transcriptional activity of estrogen receptors and/or the activation of the src family of tyrosine kinases.\",\n \"This invention also relates to novel compositions, methods, and assays for differentiating between genomic and non-genomic activity of nuclear receptors (e.g.\",\n \"estrogen receptors).\",\n \"Additionally, this invention relates to nucleic acids encoding MNAR, antisense, recombinant expression of nucleic acids, as well as host cells, compositions, and assays that utilize the nucleic acids and protein of this invention.\",\n \"BACKGROUND OF THE INVENTION Nuclear hormone receptors are a superfamily of ligand-inducible transcription factors, which, as a class, are involved in ligand-dependent transcriptional control of gene expression.\",\n \"Binding of a specific ligand, inducing conformational changes in the receptor molecule, affects receptor interaction with other transcription factors, and, ultimately, formation of the preinitiation complex.\",\n \"This process regulates the rate of gene transcription.\",\n \"(D. J. Mangelsdorf, et al., Cell 83: 835-9, 1995).\",\n \"The nuclear hormone receptor superfamily includes steroid hormone receptors, non-steroid hormone receptors, and orphan receptors.\",\n \"The receptors for glucocorticoids (GR), mineralcorticoids (MR), progestins (PR), androgens (AR), and estrogens (ER) are examples of classical steroid receptors.\",\n \"In addition to steroid hormone receptors, this nuclear hormone receptor superfamily consists of receptors for non-steroid hormones, such as vitamin D, thyroid hormones, and retinoids.\",\n \"Moreover, a range of nuclear receptor-like sequences have been identified which encode so called “orphan” receptors.\",\n \"These orphan receptors are structurally related to and, therefore, classified as nuclear hormone receptors, although no putative ligands have been identified yet.\",\n \"(Cell, 83: 851-857, 1995).\",\n \"The superfamily of nuclear hormone receptors is characterized structurally and functionally by a modular structure in comprising six distinct structural and functional domains, A to F. More specifically these receptors have a variable N-terminal region (domain A/B); followed by a centrally located, highly conserved DNA-binding domain (hereinafter referred to as DBD; domain C); a variable hinge region (domain D); a conserved ligand-binding domain (hereinafter referred to as LBD; domain E); and a variable C-terminal region (domain F).\",\n \"Cell, supra.\",\n \"The N-terminal region, which is highly variable in size and sequence, is poorly conserved among the different members of the superfamily.\",\n \"This part of the receptor is involved in the modulation of transcription activation.\",\n \"The DBD consists of approximately 66 to 70 amino acids and is responsible for DNA-binding activity: This domain targets the receptor to specific DNA sequences, called hormone responsive elements (hereinafter referred to as HRE), within the transcription control unit of specific target genes on the chromatin.\",\n \"Steroid receptors such as GR, MR, PR, and AR recognize similar HRE DNA sequences, while the ER recognizes a different DNA HRE sequence.\",\n \"After binding to DNA, the steroid receptor is thought to interact with components of the basal transcriptional machinery and with sequence-specific transcription factors, thereby modulating the expression of specific target genes.\",\n \"The LBD is located in the C-terminal part of the receptor and is primarily responsible for ligand binding activity.\",\n \"In this way, the LBD is essential for recognition and binding of the hormone ligand and, in addition, possesses a transcription activation function, thereby determining the specificity and selectivity of the hormone response of the receptor.\",\n \"Although moderately conserved in structure, the LBD's are known to vary considerably in homology among the individual members of the nuclear hormone receptor superfamily.\",\n \"When a hormone ligand for a nuclear receptor enters the cell and is recognized by the LBD, it will bind to the specific receptor protein, thereby initiating an allosteric alteration of the receptor protein.\",\n \"(Cell, supra).\",\n \"As a result of this alteration the ligand/receptor complex switches to a transcriptionally active state and, as such, is able to bind through the presence of the DBD with high affinity to the corresponding HRE on the chromatin DNA.\",\n \"In this way, the ligand/receptor complex modulates expression of the specific target genes.\",\n \"The diversity achieved by this family of receptors results from their ability to respond to different ligands.\",\n \"In addition to modulation of genomic activity, hormone receptor complexes can have important and varied non-genomic effects.\",\n \"This non-genomic activity is characterized by fast and transient increase in intracellular second messengers.\",\n \"In turn, these second messengers are involved in a number of different pathways and cascades that affect cellular functions, such as proliferation and differentiation.\",\n \"More generally, steroid hormone receptors are connected with embryonic development, adult homeostasis, and organ physiology.\",\n \"Various diseases and abnormalities are ascribed to a disturbance in steroid hormone action.\",\n \"Since steroid receptors exercise their influence as hormone-activated transcription modulators and as hormone-activated stimulators of non-genomic activity, further investigation into various approaches to modifying, interacting with, or modulating these receptors is an area of immense significance.\",\n \"For example, mutations and defects in these receptors, as well as overstimulation or blocking of these receptors, may provide better insight into the underlying mechanism of the hormone signal transduction pathway, thereby leading to increased efficacy in treatment of a wide variety of steroid receptor-linked diseases and abnormalities.\",\n \"Estrogen Receptors Estrogen (E2) exerts numerous biological effects in different tissues through an interaction with the ER.\",\n \"Amino acid sequence analyses, transient transfection studies, and mutational dissections of ER indicate that ER has the classical modular structure described above.\",\n \"The N-terminal A/B domain of the ER contains a transactivation function, referred to as transcriptional activation function 1 (TAF-1).\",\n \"The DBD contains two zinc fingers and is responsible for DNA recognition.\",\n \"The LBD and a second transactivation function, referred to as TAF-2, is located at the C-terminal of ER.\",\n \"Upon binding to hormone, the ER undergoes an activation and transformation step.\",\n \"The activated ER interacts with specific estrogen response elements (EREs) that are located in the promoter region of estrogen-regulated genes and that influence its target gene transcription.\",\n \"Over the past decade, numerous studies have provided a basic understanding of both the effects of ligand (agonist/antagonist) on the ER and the relationship between the structure and function of the ER.\",\n \"Nevertheless, little is known regarding the mechanisms for the non-genomic activity of ER.\",\n \"ERβ appears to be distinct from the more commonly known estrogen receptor, referred to as ERα.\",\n \"Collectively, ERα and ERβ are referred to herein as ER.\",\n \"The DBD of ERβ is 90% identical to that of ERα.\",\n \"However, the overall homology between the ligand binding domain (LBD) of ERα and ERβ is less than 55%.\",\n \"Like ERα, ERβ can stimulate transcription from an ERE in a ligand-dependent manner.\",\n \"It has been established that estrogens induce fast and transient increases in the levels of intracellular second messengers, including calcium and cAMP, and that estrogens induce activation of mitogen-activated protein kinase (MAPK) and phospholipase C (Collins and Webb, 1999).\",\n \"In fact, numerous studies have demonstrated that estrogens induce rapid and transient activation of the Src/Ras/MAP kinase phosphorylation pathway.\",\n \"Activation of this pathway triggers vital cellular functions including cell proliferation and differentiation.\",\n \"The time course of these acute events parallels that elicited by peptide hormones, thus supporting the hypothesis that these events do not involve the ‘classical’ genomic action of estrogens.\",\n \"Recent data also suggests a direct link between the estrogen receptor and the mitogen-activated protein (MAP) kinase-signaling cascade.\",\n \"MAP kinases are a family of serine-threonine kinases that are phosphorylated and activated in response to a variety of signals.\",\n \"These enzymes transduce extracellular signals from multiple membrane receptors to intracellular targets, including transcription factors, cytoskeletal proteins and enzymes.\",\n \"The MAP kinase family includes the extracellular-signal related kinases (ERKs), p38 and cJun N-terminal kinases which signal through a pathway involving sequential activation of Ras, Raf and mitogen-activated protein kinase (MEK).\",\n \"(S. M. Thomas, J. S. Brugge, Annual Review of Cell & Developmental Biology 13, 513-609, 1997).\",\n \"In pulmonary endothelial cells, neuronal cells, osteoblasts and osteoclasts, 17β-estradiol (E2) has been reported to rapidly activate the MAPK pathway.\",\n \"More specifically, in pulmonary endothelial cells, 17β-estradiol (E2) has been reported to rapidly stimulate nitric oxide (NO) production, thus explaining the ability of E2 to induce acute dilation of blood vessels.\",\n \"Chen et al.\",\n \"(Chen et al., 1999) have recently reported that E2 induces rapid activation of endothelial nitric oxide synthase (eNOS) in isolated pulmonary endothelial cells.\",\n \"Estrogen activation of eNOS was shown to occur through the rapid activation of the MAP kinase pathway.\",\n \"Complimentary studies have also shown that E2 induces calcium-dependent translocation of eNOS from the plasma membrane to intracellular sites close to the nucleus—an action that is rapid (within 5 minutes), receptor-mediated but nongenomic (Goetz et al., 1999).\",\n \"In addition, ligand-dependent ERα interaction with the p85α regulatory subunit of phosphatidylinositol-3-OH kinase (PI(3)K) was recently implicated in mediation of cardiovascular protective effects of estrogen (Simoncini T. and J. K., 2000).\",\n \"Stimulation with estrogen increases ERα associated PI(3)K activity, thereby leading to the activation of protein kinase B/Akt and endothelial nitric oxide synthase (eNOS).\",\n \"In neuronal cells, rapid activation of the MAP kinase signaling pathway by E2 results in neuroprotection in primary cortical neurons after glutamate excitotoxicity (Singer et al., 1999).\",\n \"These neuroprotective effects of estrogens, which were reported to occur within 5 minutes after exposure, were mediated through the transient activation of c-Src-tyrosine kinases and tyrosine phosphorylation of p21 (ras)-guanine nucleotide activating protein in an ER-dependent manner.\",\n \"Similarly, in osteoblasts E2 induces MAP kinase phosphorylation and activation within 5 minutes (Endoh et al., 1997).\",\n \"MAP kinase activation by estrogens may, therefore, regulate cell proliferation and differentiation in these cells leading to increased bone formation.\",\n \"Several studies also indicate that estrogens activate the MAP kinase pathway in osteoclasts through activation of Src kinase (Oursler, 1998).\",\n \"It has been also shown that MAP kinase activation, could be involved in the regulation of enzymes needed for bone resorption (Kristen D. Brubaker, 1999).\",\n \"In the human mammary cancer derived cell lines, MCF-7 and T47D, as well as in the human colon cancer derived cell line, Caco-2, E2 activates the signal transducing Src/Ras/Erk pathway (Migliaccio et al., 1993), (Migliaccio et al., 1996), (Migliaccio et al., 1998), (Migliaccio et al., 2000).\",\n \"This activation is mediated by direct ER-Src interaction.\",\n \"Interestingly, progesterone also activates the same pathway in T47D cells (Migliaccio et al., 1998).\",\n \"The Src/Ras/ERk signaling pathway is a well-known target of growth factors.\",\n \"Importantly, activation of this pathway requires direct interaction of ER with Src.\",\n \"Activation of this pathway triggers different cellular responses such as proliferation or differentiation [Cantley, 1991]; [Marshall, 1996]; [Downward, 1997].\",\n \"Its activation by estrogen receptor ligands, such as steroid hormones, explicates their involvement in the cell cycle control.\",\n \"These data support the view that non-transcriptional/nongenomic activity of ER may be responsible for stimulation of cell growth.\",\n \"However, the details of the molecular mechanism of this process are largely unknown.\",\n \"Src activation has been previously observed in a large number of human breast and colon carcinomas ([Rosen, 1986]; [Ottenhoff-Kalff, 1992]).\",\n \"Activation of Src induces mammary tumors in transgenic mice ([Guy, 1994]).\",\n \"Constitutively active Ras mutants have been found in 25-30% of all cancers, including breast cancer ([Kasid, 1987]).\",\n \"Recent studies have suggested the existence of a plasma membrane estrogen receptor unrelated to the classical ER.\",\n \"However, cloning or isolation of this membrane ER has not been accomplished, while others have suggested that a subpopulation of the classical ER is associated with the cell membrane and is responsible for the rapid effects of estrogens.\",\n \"Transcriptional activity of nuclear receptors itself is a target for tight regulation by multiple signaling pathways, including those stimulated by the neurotransmitter dopamine ([Power, 1991]; [Smith, 1993]), growth factors such as epidermal growth factor (EGF), transforming growth factor-α (TGF-a) and insulin-like growth factor I (IGF-I) ([Ignar-Trowbridge, 1996], [Aronica, 1993]; [Bunone, 1996]), and by activators of protein kinase C ([Aronica, 1994]).\",\n \"The molecular mechanisms for such cross coupling are thought to be mediated, at least in part, by receptor phosphorylation.\",\n \"The ER has been demonstrated to be phosphorylated by MAP kinase in response to EGF, thereby resulting in stimulation of AF1 ([Kato, 1996]; [Bunone, 1996]).\",\n \"Other studies have demonstrated that the ER ([Auricchio, 1987]; [Arnold, 1995]; [Pietras, 1995]), Thyroid hormone receptor (TRβ) ([Lin, 1992]), RARγ ([Rochette-Egly, 1992]), glucocorticoid receptor ([Rao, 1987]) and the orphan receptor HNF-4 ([Ktistaki, 1995]) are also targets for tyrosine phosphorylation.\",\n \"A specific tyrosine phosphorylation site has been identified within the AF2 domain of the human ER (hER) at amino acid 537 (Y537) ([Castoria, 1993]; [Arnold, 1995]).\",\n \"This tyrosine is located immediately N-terminal of the AF2 sequence and is conserved in all known ER sequences from diverse species including the ERβ ([Kuiper, 1996]; [Mosselman, 1996]).\",\n \"Although the molecular mechanisms of non-genomic and genomic activities of E2 are not well understood, the present invention should allow us to create a new generation of therapeutics with tissue and gene selective activity.\",\n \"To better understand the tissue selective action of some ER ligands, we evaluated the expression and activity of ER-interacting proteins in different cell lines.\",\n \"Using affinity purification and mass spectrometer based microsequencing, we discovered a novel ER interacting protein, designated as MNAR (modulator of non-genomic activity of nuclear receptors).\",\n \"SUMMARY OF THE INVENTION The present invention provides for a novel protein modulator of non-genomic activity of nuclear receptors (MNAR) and nucleic acids that encode such proteins.\",\n \"The MNAR protein can interact with nuclear hormone receptors, such as estrogen receptors.\",\n \"In general, the protein or polypeptide of MNAR refers to the amino acid sequence of an MNAR.\",\n \"In particularly preferred embodiments, MNAR protein interacts with ERα and ERβ.\",\n \"This interaction is ligand dependent.\",\n \"MNAR will also form a complex with ER and kinases, such as members of the Src family.\",\n \"Src and MAP kinase activation leads to enhancement of ER transcriptional activity.\",\n \"The present invention provides (a) a novel nucleic acid comprising a polynucleotide encoding an MNAR having a putative amino acid sequence of SEQ ID No: 2 (see FIG.\",\n \"12).\",\n \"A nucleotide sequence of human MNAR is shown in SEQ ID No: 1 (See FIG.\",\n \"15).\",\n \"An amino acid sequence of human MNAR is shown in SEQ ID NO: 13 (See FIG.\",\n \"16).\",\n \"A nucleotide sequence of murine MNAR is shown in SEQ ID NO: 12 (See FIG.\",\n \"17).\",\n \"An amino acid sequence of murine MNAR is shown in SEQ ID NO: 13 (See FIG.\",\n \"18).\",\n \"This invention provides novel isolated nucleic acid comprising a polynucleotide selected from the group consisting of: (a) a polynucleotide encoding a novel protein modulator of non-genomic activity of nuclear receptors (MNAR) comprising the amino acid sequence of SEQ ID NO:2 or of SEQ ID NO:13; (b) a polynucleotide that hybridizes under highly stringent conditions with (i) a region of the nucleotide sequence of SEQ ID NO: 1 or of SEQ ID NO:12, (ii) a subsequence of at least 100 nucleotides of the nucleotide sequence of SEQ ID NO: 1 or of SEQ ID NO:12, (iii) or a complementary strand of (i) or (ii); (c) a polynucleotide comprising a sequence with at least 85% identity to a polynucleotide coding sequence of SEQ ID NO: 1 or of SEQ ID NO: 12; (d) a variant of the polynucleotide comprising a polynucleotide coding sequence of SEQ ID NO:1 or of SEQ ID NO:12; and (e) a polynucleotide encoding a polypeptide fragment comprising the amino acids of SEQ ID NOS 20 or 23 (f) a complement or antisense form of the polynucleotide of (a), (b), (c), (d) or (e); wherein the polynucleotide does not encode a p160 protein (FIG.\",\n \"1).\",\n \"In preferred embodiments the isolated nucleic acid comprises an allelic variant of the polynucleotide.\",\n \"In certain embodiments, the nucleic acid is operably linked to an expression control sequence (e.g.\",\n \"heterologous or homologous control sequences).\",\n \"This invention provides for a host cell, including bacteria, yeast, insect, and mammalian cells transformed with such isolated nucleic acids.\",\n \"This invention also relates to a polypeptides comprising an amino acid sequence selected from the group consisting of: (a) an amino acid sequence for a protein modulator of non-genomic activity of nuclear receptors (MNAR) comprising the amino acid sequence of SEQ ID NO: 2 or of SEQ ID NO:13; (b) an amino acid sequence for an MNAR fragment comprising the amino acid sequence of SEQ ID NOS:14, 15 or 16; (c) a variant of (a) or (b), and (d) a fragment of (a), (b), or (c).\",\n \"In alternative embodiments, these polypeptides comprise an amino acid sequences which is more than 90%, or 95% identical to the amino acid sequence of SEQ ID NO:13 or 14.In particular embodiments, the polypeptides of the invention comprise the amino acid sequence of SEQ ID NOS 2, 13, 14, 15 16 20 or 22.In additional embodiments the polypeptides of this invention can stimulates the activity of a tyrosine kinase in the presence of one or more nuclear receptors.\",\n \"The nuclear receptors can be selected from steroid and non-steroid nuclear receptors as well as orphan nuclear receptors.\",\n \"In certain embodiments, the nuclear receptor is an estrogen receptor.\",\n \"Alternative embodiments involve monoclonal and polyclonal antibodies which correspond to MNAR polypeptides as well as fragments thereof.\",\n \"In view of the modulation by MNAR of nuclear receptors, many embodiments of this invention relate to modulating the transcriptional activity of a nuclear receptor, comprising providing to the loci of a nuclear receptor an MNAR polypeptide.\",\n \"For example, one can modulate the transcriptional activity of a nuclear receptor, by (a) transforming a host cell with the recombinant vector having the nucleic acid sequence for MNAR according to claim 1; and (b) culturing said transformed host cell to express the nucleic acid in the presence of a nuclear receptor.\",\n \"This invention also presents numerous approaches for identifying compounds that affect non-genomic activity or genomic activity of nuclear receptors.\",\n \"In one embodiment, a method of identifying compounds exhibiting non-genomic activity versus genomic activity compries administering a test compound to a cell comprising an MNAR-nuclear receptor complex and measuring the non-genomic or genomic activity resulting from the test compound.\",\n \"In these methods, preferably, at least one ligand of a nuclear receptor is present in the cell, or a kinase is present, or both are present.\",\n \"Other methods are directed to the screening for a compound that modulates the activity of MNAR on a nuclear receptor, comprising the steps of (a) contacting a test compound with an MNAR polypeptide; and (b) determining whether said test compound specifically binds said polypeptide.\",\n \"In addition, one can conduct a method of screening for a compound that modulates the activity of MNAR on a nuclear receptor, said method comprising the steps of (a) adding a test compound to a cell comprising the polypeptide of and a nuclear receptor; and (b) comparing the MNAR activity before and after said adding step.\",\n \"Further approaches to this method involve adding a test compound to a control comprising a mutant cell lacking MNAR activity or with significantly reduced MNAR activity.\",\n \"In each of these methods one can use a control to assess the non-genomic activity (or genomic activity) of a compound by administering a compound to a cell in the presence of MNAR and a nuclear receptor, and then repeating the experiment in the absence of MNAR and comparing the level of non-genomic activity.\",\n \"Preferred methods employ a cell overexpresses MNAR a nuclear receptor or both.\",\n \"The genomic activity of a compound is detected by having the nuclear receptor operatively associated with a reporter.\",\n \"Certain embodiments of the present invention also relate to a method of identifying compounds having selective genomic versus non-genomic activity said method comprising the steps of (a) adding a test compound to a cell comprising a MNAR-nuclear receptor complex; and (b) comparing the genomic versus non-genomic activity before and after said adding the test compound.\",\n \"A selective genomic activity can be measured by conventional means.\",\n \"Preferably, the increase or positive effect of non-genomic activity, as measured, is a two-fold increase after addition of said test compound to a cell in the presence of MNAR-nuclear receptor when compared to genomic activity with test compound in the absence of MNAR and wherein no change is observed in non-genomic activity after addition of said test compound.\",\n \"On the other hand, when selecting for test compounds for non-genomic activity, there should be at least about a two-fold increase after addition of said test compound to a cell in the presence of MNAR-nuclear receptor complex when compared to non-genomic activity with test compound in the absence of MNAR and wherein no change is observed in genomic activity after addition of said test compound.\",\n \"One can measure the effect by determining an increase or decrease in the transcriptional activity of a nuclear receptor in the presence of MNAR.\",\n \"In a variety of cells, healthy or diseased, it can be valuable to understand whether a beneficial or healthy state for a cell results from non-genomic or genomic activity.\",\n \"The present invention permits such determinations by comparing a desired phenotype of a cell, in the presence and absence of MNAR.\",\n \"The present invention also relates to non-human transgenic mammals, wherein one or more cells comprise at least one non-functional endogenous MNAR polynucleotide sequence, at least one non-functional endogenous nuclear receptor or both.\",\n \"In preferred embodiments, the non-human transgenic mammal do not have functional endogenous forms of ERa and ERA.\",\n \"Alternatively, transgenic mammal comprises substitution of an inducible/repressable promoter for the endogenous MNAR promoter.\",\n \"The non-human transgenic mammal of claim may further comprise an exogenous selectable marker gene under the control of a promoter active in at least one cell type of said mammal.\",\n \"This invention also relates to a method of screening for a compound that disrupts the function of MNAR, comprising using a protein-protein interaction assay to identify compounds that interact with MNAR.\",\n \"Once can employ any of the well-known two-hybrid assays or other conventional assays to study protein-protein interactions and the disruption thereof by test compounds, which can be a natural product isolate, small molecule chemical, peptide, etc.\",\n \"MNAR protein and MNAR encoding nucleic acids can be obtained from mammalian cells and tissues (e.g.\",\n \"human and murine cells and tissues).\",\n \"Recombinant MNAR and anti-MNAR antibodies find use in drug screening, diagnostics, and therapeutics.\",\n \"In particular, the MNAR provides valuable reagents in developing specific biochemical assays for screening compounds for ER ligands that agonize or antagonize the enzymatic activity of the Src family of tyrosine kinases, which are involved in regulating gene expression.\",\n \"Additionally, the MNAR provides valuable reagents in developing specific biochemical assays for screening compounds for a variety of nuclear receptor ligands.\",\n \"Accordingly, one can analyze the effects on any pathway involving a nuclear receptor, such as ER, and/or a kinase.\",\n \"For example, one can use MNAR to set up assays to determine the effect of ER ligands or test compounds on (I) cell cycle stimulation in cancer cells; (II) cell proliferation, cell differentiation and cell conservation for bone development or modulation; (III) neuroprotective activity of ER ligands/test compounds in nerve cells; and (IV) pulmonary characteristics of endothelial cells.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS FIG.\",\n \"1 is an alignment comparing the amino acid sequence of MNAR and p160, a protein homologous to MNAR.\",\n \"FIG.\",\n \"2 shows estrogen receptor β interaction with MNAR.\",\n \"Panel A presents a silver stained gel obtained by GST-ERβ-LBD pull-down from MCF-7 cells extract.\",\n \"The resulting peptides were purified using reverse phase HPLC column and identified by mass spectrometry based microsequencing.\",\n \"The sequences of these peptides are provided in panel B.\",\n \"FIG.\",\n \"3.depicts the results of the expression analysis of MNAR in various cell lines, as evaluated by Northern blotting from experiments in Example 2.FIG.\",\n \"4 (A and B) shows the results of MNAR expression analysis in various tissues, as evaluated by Northern blotting from experiments in Example 2.FIGS.\",\n \"5A and 5B show analysis of flag-tagged MNAR interaction with ERα and ERβ, or receptors liganded with E2, or 4-OH tamoxifen.\",\n \"Also shown are the results of full-length flag-MNAR interaction with transcribed/translated androgen (AR) and glucocorticoid receptors (GR) with and without their corresponding ligands (FIGS.\",\n \"5C and 5D).\",\n \"FIG.\",\n \"6A depicts the results of the experiment of example 4, wherein the tests are conducted to evaluate the in vitro interaction of Src or Lck with MNAR and/or ERa in the presence and in the absence of 17β-estradiol.\",\n \"FIG.\",\n \"6B depicts the results of the experiment of example 4, wherein the tests are conducted to evaluate the in vitro interaction of Src or Lck with MNAR and/or ERβ in the presence and in the absence of 17β-estradiol.\",\n \"FIG.\",\n \"7 is a schematic depiction of the proposed model of ER-MNAR-Src interaction.\",\n \"FIG.\",\n \"8 is an SDS-PAGE gel analysis of the ER-MNARc complex on cSrc enzymatic activity.\",\n \"cSrc enzymatic activity was evaluated in the absence (lanes 1 and 2) or presence of ER (lanes 3-6), E2 (lanes 3 and 5), and MNAR (lanes 2, 5, and 6) using acid denatured enolase as a substrate.\",\n \"Data presented in FIG.\",\n \"8 indicate that MNAR itself strongly stimulates cSrc enzymatic activity.\",\n \"FIG.\",\n \"9 is a series of graphs depicting the results from the experiments of example 7, wherein the tests are conducted to evaluate whether MNAR affects ER-mediated transcription.\",\n \"FIG.\",\n \"9A shows the result for ERα, MNAR, and/or SRC3.FIG.\",\n \"9B depicts the results for ERα and MNAR when cells are treated with E2, E2 plus PP2, or E2 plus PD98059.FIG.\",\n \"9C shows results of MCF-7 cells treated with antisense (AS), E2, E2 plus control, or E2 plus antisense.\",\n \"A primer/probe set is designed to target MNAR, PS2, and cathepsin D, respectively.\",\n \"FIG.\",\n \"9D shows results of a Western blot analysis of MCF-7 cell extracts transfected with antisense or reverse control oligomers stimulated with E2 with rabbit polyclonal MNAR antiserum.\",\n \"FIG.\",\n \"10 depicts an SDS-gel analysis of MCF-7 cells untransfected and/or transfected with a Flag-MNAR expression vector untreated and treated with ±2.Flag-MNAR (panel A), ERα (panel C and D), and cSrc (panel B) were immunoprecipitated from the cell extracts using their corresponding antibodies.\",\n \"Material precipitated using ERα antiserum was probed with rabbit polyclonal anti-MNAR antibody (panel E).\",\n \"E2-enhancement of ERα-MNAR interaction was clearly detected.\",\n \"FIG.\",\n \"11 depicts the results of the analysis of MNAR on E2 induced Erk activation, as evaluated by Western blotting from experiments in Example 9.FIG.\",\n \"12 is a schematic depiction of the structural-functional organization of MNAR and 13 MNAR mutants (mutants 2-14), which were generated and tested for their ability to enhance ER-stimulated transcription when transfected into HepG2 cells under the experiments of Example 10.FIG.\",\n \"13 is a bar graph depicting the results of an analysis of MNAR deletion mutants (mutants 2-14), which were tested for their ability to enhance ER-stimulated transcription when transfected into hepG2 cells in the experiments of Example 10.FIG.\",\n \"14 depicts is a bar graph depicting the results of an analysis of PXXP motif MNAR mutants, which were tested for their ability to activate ER transcriptional activity When transfected into HepG2 cells in the experiments of Example 10.FIG.\",\n \"15 depicts a polynucleotide sequence of human MNAR (SEQ ID NO:1).\",\n \"FIG.\",\n \"16 depicts an amino acid sequence of human MNAR (SEQ ID NO: 2).\",\n \"FIG.\",\n \"17 depicts a polynucleotide sequence of murine MNAR (SEQ ID NO:12)).\",\n \"FIG.\",\n \"18 depicts an amino acid sequence of murine MNAR (SEQ ID NO:13).\",\n \"DESCRIPTION OF THE SPECIFIC EMBODIMENTS Definitions Functional analogs of a given MNAR protein or protein with “substantial functional identity” to a given MNAR are defined as proteins that exhibit one or more biochemical properties specific to such MNAR.\",\n \"“Nucleic acid molecule” means a single-stranded or double stranded molecule and includes RNA molecules, DNA molecules, and analogs of RNA or DNA molecules.\",\n \"“Modulating transcription” means altering transcription, and includes changing the rate of transcription initiation, the level of transcription, or the responsiveness of transcription/transcription initiation to regulatory controls.\",\n \"“Neoplastic state of cells” means any new growth of cells which may be benign or malignant.\",\n \"The terms “substantially pure” or “isolated” mean that the MNAR protein, MNAR protein fragment, or nucleic acid encoding a MNAR or MNAR fragment are unaccompanied by at least some of the material with which it is normally associated in its natural state.\",\n \"A composition of a substantially pure MNAR or portion thereof may contain excipients and additives useful in diagnostic, therapeutic and investigative reagents.\",\n \"“Altered expression state” means that the expression of MNAR in a naturally-occurring state is altered to increase or decrease MNAR expression.\",\n \"In comparison to the naturally-occurring state.\",\n \"“Substantial sequence identity” means that a portion of the protein or nucleic acid presents at least about 70%, more preferably at least about 80%, and most preferably at least about 90%, 95%, or 99% sequence identity with a MNAR sequence portion.\",\n \"Where the sequence diverges from native MNAR sequences disclosed herein, the differences are preferably conservative, i.e., an acidic for an acidic amino acid substitution or a nucleotide change providing a redundant codon.\",\n \"Dissimilar sequences are typically aggregated within regions rather than being distributed evenly over the polymer.\",\n \"A substantially identical sequence hybridizes to a complementary MNAR-encoding sequence under relatively high stringency.\",\n \"For the purposes of defining high stringency southern hybridization conditions, reference can conveniently be made to Sambrook et al.\",\n \"(1989) at pp.\",\n \"387-389 which is herein incorporated by reference where the washing step at paragraph 11 is considered high stringency.\",\n \"“Fusion protein or polypeptide” means a protein or polypeptide that comprises at least a first amino acid sequence fused to a second amino acid sequence, where such fusion does not occur in a naturally occurring protein or polypeptide.\",\n \"Preferably, at least one amino acid sequence in the fusion protein or polypeptide is that of MNAR.\",\n \"“Mutant or mutation” means any detectable change in genetic material, e.g.\",\n \"DNA, or any process, mechanism or result of such a change.\",\n \"This includes gene mutations, in which the structure (e.g.\",\n \"DNA sequence) of a gene is altered, any gene or DNA arising from any mutation process, and any expression product (e.g.\",\n \"protein, polypeptide, or enzyme) expressed by a modified gene or DNA sequence.\",\n \"“Variant” is used to indicate a modified or altered gene, DNA sequence, protein, polypeptide, enzyme, cell, etc., i.e.\",\n \"any kind of mutant.\",\n \"A variant of a naturally occurring protein or polypeptide, for example, comprises a substitution, deletion, modification, and/or insertion of one or more amino acids in the naturally occurring protein or polypeptide.\",\n \"“Recombinant” means any genetic material (e.g.\",\n \"DNA sequence) resulting from the insertion into the sequence or chain, by biological or chemical means, of a sequence (e.g.\",\n \"a full-length or partial DNA sequence) not originally biologically or naturally present in that genetic material (e.g.\",\n \"DNA sequence).\",\n \"“Protein or polypeptide fragment” means any smaller portion of a larger protein or polypeptide, where preferably the fragment has at least 8 amino acids and more preferably, 25, 50, 100 or 200 amino acids.\",\n \"Non-genomic activity of a receptor is “beneficial to a cell” wherein the cell exhibits presence of a desired activity or phenotype upon introduction of increased amount of MNAR to the cell.\",\n \"Non-genomic activity of a receptor is “not beneficial to a cell” wherein the cell exhibits absence of a desired activity or phenotype upon introduction of increased amount of MNAR to the cell.\",\n \"MNAR Characterization Sequence analysis MNAR revealed multiple LXXLL motifs localized in N-terminal portion of MNAR′ molecule.\",\n \"Similar motifs in other transcription factors were previously shown to form contacts with a hydrophobic groove on the surface of the ligand binding domains of nuclear receptors ((Heery et al., 1997), (Torchia et al., 1997)).\",\n \"Another important feature of the MNAR molecule is a proline-rich domain.\",\n \"This domain can be utilized for interaction with SH3 domains of the Src family of protein kinases.\",\n \"Initially we hypothesized that MNAR may potentiate nuclear receptors interaction with Src kinases.\",\n \"Our data indicate that estrogen receptors α and β interact with MNAR, and this interaction is enhanced by 17β-estradiol but not by 4(OH) tamoxifen.\",\n \"Interestingly, the MNAR-ER complex, both in-vitro and in-vivo strongly interacts and stimulates enzymatic activity of Src kinases.\",\n \"Importantly, overexpression of MNAR leads to enhancement of ER transcriptional activity.\",\n \"We expect that this activation of ER is due to its phosphorylation by one of the kinases downstream of Src.\",\n \"Accordingly, we submit that MNAR modulates ER interaction with Src kinases and by doing so mediates the nongenomic activity of ER.\",\n \"Our data supports the development of experimental models that will allow us to assay for ER ligands that are able to differentiate between genomic and non-genomic activity of ER ligands.\",\n \"Ligands that do not affect transcriptional activity of ER, but stimulate the Src enzymatic activity may possess important bone-sparing, CNS protective, and cardioprotective activity.\",\n \"On the other hand, compounds that selectively control transcriptional activity of ER would have a much more selective action, thereby avoiding multiple side affects.\",\n \"Overall, our ability to separate genomic and nongenomic activity of nuclear receptor ligands, particularly ER ligands, may allow us to create a new generation of therapeutics with profoundly focused, selective action.\",\n \"Our data also support the development of experimental models that will allow us to assay for nuclear receptor ligands that are able to differentiate between genomic and non-genomic activity of nuclear receptor ligands.\",\n \"Ligands that do not affect transcriptional activity of various nuclear receptors, but stimulate the Src enzymatic activity or other non-genomic enzymatic activity may possess important therapeutic properties or activities.\",\n \"On the other hand, compounds that selectively control transcriptional activity of nuclear receptors would have a much more selective action, thereby avoiding multiple side affects.\",\n \"Overall, our ability to separate genomic and non-genomic activity of nuclear receptor ligands may allow us to create a new generation of therapeutics with profoundly focused, selective action.\",\n \"Nucleic Acid Sequences Encoding MNAR and Corresponding MNAR Proteins A biologically active MNAR, or MNAR fragment thereof, retains one or more of the MNAR's functions, such as the ability to specifically form a complex with ER, or to modulate or enhance the transcriptional activity of ER.\",\n \"Exemplary assays for biological activity are described below and in the working exemplification.\",\n \"Specific binding is empirically determined by contacting, for example a MNAR, with a mixture of components and identifying those components that preferentially bind the MNAR.\",\n \"Specific binding may be conveniently shown by a number of methods, including but not limited to yeast and mammalian hybrid systems and competitive binding studies.\",\n \"For example, a human cDNA library was screened using the yeast two-hybrid assay to identify potential coactivators, receptors and/or ligands for ER or MNAR.\",\n \"In addition to the analysis of the interactions between kinases and the MNAR-ER complex, MNAR can be used in similar strategies to assess the interaction between ER ligands and ER with other known transcription factors, including other nuclear receptors, and to identify novel interacting proteins.\",\n \"For example, since the MNAR-ER interaction can be enhanced by 17β-estradiol, one can assess the effect of test agents, which may be compounds or proteins or peptides.\",\n \"Furthermore, the yeast two-hybrid system, the mammalian two-hybrid system, surface plasmon resonance assays, immunoprecipitation assays, or other assays can be used to demonstrate a direct interaction between MNAR and ER in the presence or absence of Src.\",\n \"Similarly, these assay systems could be used to broadly screen for known and novel proteins that interact with MNAR-ER complex or MNAR-ER-Src complex.\",\n \"The invention provides recombinantly produced MNAR proteins, MNAR analogs and fragments thereof.\",\n \"These recombinant products are readily modified through physical, chemical, and molecular techniques disclosed or cited herein or otherwise known to those skilled in the relevant art.\",\n \"According to a particular embodiment of the invention, fragments of the MNAR-encoding sequences are spliced with heterologous sequences to produce fusion proteins.\",\n \"Such fusion proteins find particular use in modulating ER activity in vitro and in vivo.\",\n \"MNAR can be further modified by methods known in the art.\",\n \"For example, MNAR may be phosphorylated or dephosphorylated, glycosylated or deglycosylated, with or without radioactive labeling, etc.\",\n \"Serine, threonine, and tyrosine residues can particularly provide useful phosphorylation sites.\",\n \"Especially useful are modifications that alter MNAR solubility, membrane transportability, stability, and binding specificity and affinity.\",\n \"Some examples include fatty acid-acylation, proteolysis, and mutations in nuclear receptor transcription factor interaction domains that stabilize binding.\",\n \"Particularly, one may modify the proline-rich region of one of the LXXLL motifs.\",\n \"A substantially pure or isolated MNAR protein, or MNAR portion encoded by nucleic acid, is generally at least about 1% homologous to said MNAR encoding nucleic acid; preferably at least about 10%; more preferably at least about 50%; and most preferably at least 90%.\",\n \"Nucleic acid weight percentages are determined by dividing the weight of the MNAR or MNAR portion encoding nucleic acid, including alternative forms and analogs such as alternatively spliced or partially transcribed forms, by the total nucleic acid weight present.\",\n \"The invention also provides for MNAR sequences modified by transitions, transversions, deletions, insertions, or other modifications such as alternative splicing and such alternative forms, genomic MNAR sequences, MNAR gene flanking sequences, including MNAR regulatory sequences and other non-transcribed MNAR sequences, MNAR mRNA sequences, and RNA and DNA antisense sequences complementary to MNAR encoding sequences, sequences encoding xenogeneic MNARs and MNAR sequences comprising synthetic nucleotides, e.g., the oxygen of phosphate group may be replaced with sulfur, methyl, or the like.\",\n \"The modified MNAR encoding sequences or related sequences encode proteins with MNAR-like functions.\",\n \"There will generally be substantial sequence identity between at least a portion thereof and a portion of a MNAR, preferably at least about 40%, more preferably at least 80%, most preferably at least 90%, 95%, or 99%, particularly conservative substitutions, particularly within the proline-rich regions and LXXLL motifs and regions encoding protein domains involved in protein-protein interactions, particularly MNAR-ER or MNAR-ER-kinase interactions, particularly MNAR-nuclear receptor or MNAR-nuclear receptor-kinase interactions.\",\n \"MNAR can be subject to alternative purification, biosynthesis, modification or use by methods disclosed herein or otherwise known in the art.\",\n \"For example, the amino acids can be modified in a number of ways, including but not limited to, altering stability, solubility, binding affinity, specificity, and methylation.\",\n \"The amino acid sequences of the present invention may also be modified with a label capable of providing a detectable signal, either directly or indirectly.\",\n \"Exemplary labels include, but are not limited to, radioisotopes, fluorescers, and biotinylation.\",\n \"Nucleic acids encoding at least a portion of an MNAR are used to identify nuclear factors and agents which interact with that MNAR using expression screening in yeast or mammalian cells as described in Current Protocols in Molecular Biology (See “Protein Expression” in Chapter 16, 2002).\",\n \"One method of identifying these nuclear factors and agents, which interact with MNAR, is to utilize a mammalian, or yeast, two-hybrid or three-hybrid assay.\",\n \"(See, e.g., Fields S. et al., (1994) Trends Genet.\",\n \"10:286-292; Young P. et al.\",\n \"(1992) Current Biology 3:408-420; Zervos et al.\",\n \"(1993) Cell 72:223-232; Madura et al.\",\n \"(1993) J. Biol.\",\n \"Chem.\",\n \"268:12046-12054; Bartel et al (1993) Biotechniques 14:920-924; and Iwabuchi et al.\",\n \"(1993) Oncogene 8:1693-1696).\",\n \"As exemplified herein, a yeast cDNA library containing fusion genes of cDNA joined with DNA encoding the activation domain of a transcription factor (e.g.\",\n \"Gal4) are transfected with fusion genes encoding a portion of a MNAR and the DNA binding domain of a transcription factor.\",\n \"Clones encoding MNAR binding proteins provide for the complementation of the transcription factor and are identified through transcription of a reporter gene.\",\n \"See, e.g.\",\n \"Fields and Song (1989) Nature 340, 245-246 and Chien et al.\",\n \"(1991) Proc.\",\n \"Natl.\",\n \"Acad.\",\n \"Sci USA 88, 9578-9582.The invention also provides vectors comprising nucleic acids encoding MNAR or portion or analog thereof and optionally, encoding ERs and or kinases.\",\n \"A large number of vectors, including plasmid and viral vectors, have been described for expression in a variety of eukaryotic and prokaryotic hosts.\",\n \"Vectors will often include one or more replication systems for cloning or expression, one or more markers for selection in the host, e.g., antibiotic resistance, and one or more expression cassettes.\",\n \"The inserted MNAR coding sequences may be synthesized, isolated form natural sources, prepared as hybrids, etc.\",\n \"Ligation of the coding sequences to the transcriptional regulatory sequences may be achieved by known methods.\",\n \"Advantageously, vectors may also include a promoter operably linked to the MNAR encoding portion.\",\n \"Suitable host cells may be transformed/transfected/infected by any suitable method including electroporation, CaCl2 mediated DNA uptake, viral infection, microinjection, microprojectile, or other established methods.\",\n \"Alternatively, nucleic acids encoding one or more MNARs may be introduced into cells by recombination events.\",\n \"For example, a sequence can be microinjected into a cell, and thereby effect homologous recombination at the site of an endogenous gene encoding a MNAR, an analog or pseudogene thereof, or a sequence with substantial identity to a MNAR-encoding gene.\",\n \"Other recombination based methods such as non-homologous recombinations, deletion of endogenous gene by homologous recombination, especially in pluripotent cells, etc., provide additional applications.\",\n \"Appropriate host cells include bacteria, archebacteria, fungi, especially yeast, and plant and animal cells, especially mammalian cells.\",\n \"Of particular interest are E. coli, B. subtilis, Saccharomyces cerevisiae, A549 cells, CHO, COS, HeLa, Hep2, and bone cells and immortalized mammalian myeloid and lymphoid cell lines.\",\n \"Preferred host cell lines include but are not limited to human cancer cell lines such as MCF-7 and T47D cells.\",\n \"Ideally, such expression systems utilize inducible expression strategies like the TET ON/OFF system that is commercially available.\",\n \"Such cell lines are useful to define the role of MNAR in non-genomic ER activity and to assess the effect of potential compounds that modulate MNAR-ER interactions.\",\n \"A large number of transcription initiation and termination regulatory elements/regions have been isolated and shown to be effective in the transcription and translation of heterologous proteins in the various hosts.\",\n \"Examples of these regions, methods of isolation, manner of manipulation, etc.\",\n \"are known in the art.\",\n \"Suitable expression control sequences and host cell/cloning vehicle combinations are well known, and are described in Sambrook et al.\",\n \"(1989).\",\n \"MNAR encoding oligonucleotides can also be used to identify other MNARs or transcription factor coactivators.\",\n \"For example, 32P-labeled MNAR encoding nucleic acids are used to screen cDNA libraries at low stringency to identify similar cDNAs that encode proteins with MNAR related domains.\",\n \"Additionally, MNAR related proteins are isolated by PCR amplification with degenerate oligonucleotide probes using the sequences disclosed herein.\",\n \"Other experimental methods for cloning MNAR are also set out in the working exemplification below.\",\n \"Other useful cloning, expression, and genetic manipulation techniques for practicing the inventions disclosed herein are known to those skilled in the art.\",\n \"The compositions and methods disclosed herein may be used to effect gene therapy.\",\n \"See, e.g., Gutierrez et al.\",\n \"(1992) Lancet 339, 715-721.For example, cells are transfected with MNAR sequences operably linked to gene regulatory sequences capable of effecting altered MNAR expression or regulation.\",\n \"To modulate MNAR translation, cells may be transfected with MNAR complementary antisense polynucleotide.\",\n \"Antisense modulation may employ MNAR antisense sequences operably linked to gene regulatory sequences.\",\n \"An antisense oligonucleotide can be, for example, 8, 10, 15, 20, 25, 30, 35, 40, 45 or 50 nucleotide sequences in length.\",\n \"An antisense nucleic acid can be constructed using chemical synthesis and enzymatic ligation reactions using known procedures in the art.\",\n \"For example, an antisense nucleic acid (e.g.\",\n \"antisense oligonucleotide) can be chemically synthesized using naturally occurring nucleotides or various modified nucleotides designed to increase the biological activity of the molecules or to increase the physical stability of the duplex formed between antisense and sense nucleic acids, e.g.\",\n \"phosphorothiolate derivatives and acridine substituted nucleotides can be used.\",\n \"Alternatively, the antisense nucleic acid can be produced biologically using an expression vector into which a nucleic acid has been subcloned in an antisense orientation (i.e.\",\n \"RNA transcribed from the inserted nucleic acid will be of an antisense orientation to a target nucleic acid of interest.\",\n \"In the present invention, cells are transfected with a vector comprising a MNAR sequence with a promoter sequence oriented such that transcription of the gene yields an antisense transcript capable of binding to MNAR encoding mRNA.\",\n \"Transcription may be constitutive or inducible and the vector may provide for stable extrachromosomal maintenance or integration.\",\n \"Alternatively, single-stranded antisense nucleic acid sequences that bind to genomic DNA or mRNA encoding at least a portion of MNAR may be administered to the target cell at a concentration that results in a substantial reduction in MNAR expression.\",\n \"Assays for Identifying Modulators of Non-Genomic Activity of Nuclear Receptors and Therapeutic Agents The invention provides methods and compositions for identifying agents useful in modulating the genomic or non-genomic activity of nuclear receptors (e.g.\",\n \"ER).\",\n \"Such agents find use in the diagnosis or treatment of broad range of disease including, but not limited to, cancer, cardiovascular diseases, microbial and fungal infections and particularly immune disease, bone protection, etc.\",\n \"The ability to develop rapid and convenient high-throughput biochemical assays for screening compounds that work through nuclear receptor (e.g.\",\n \"ER) non-genomic activity versus nuclear receptor (e.g.\",\n \"ER) genomic activity in human cells opens a new avenue for drug development.\",\n \"In addition, this invention provides a tool for identifying whether disease states within a cell or beneficial cell states of a cell are effected by nuclear receptor (e.g.\",\n \"ER) genomic/non-genomic activities/pathways, i.e.\",\n \"one can determine whether cancer cells proliferate when nuclear receptor (e.g.\",\n \"ER) non-genomic activity or nuclear receptor (e.g.\",\n \"ER) genomic activity is decreased/enhanced.\",\n \"In general, this invention provides for categorizing many cell types as requiring nuclear receptor (e.g.\",\n \"ER) genomic/non-genomic activity.\",\n \"Common assays include yeast two-hybrid assays as disclosed by Young and Ozenberger in U.S. Pat.\",\n \"No.\",\n \"5,989,808, which is herein incorporated by reference.\",\n \"The two hybrid assays can be used to screen for small molecule compounds or peptides or proteins of interest.\",\n \"Typically, prospective agents are screened from large libraries of synthetic or natural compounds.\",\n \"For example, numerous means are available for random and directed synthesis of saccharide, peptide, and nucleic acid based compounds, see, e.g.\",\n \"Lam et al.\",\n \"(1991) Nature 354, 82-86.Alternatively, libraries and compounds are readily modified through conventional chemical, physical, and biochemical means.\",\n \"Examples of such modifications are disclosed herein.\",\n \"Useful agents are identified with a range of assays employing MNARs or MNAR encoding nucleic acids.\",\n \"The assays may also employ one or more selected from nuclear receptors, steroid receptors, orphan receptors, ER, ERE (estrogen response elements), ER ligands, kinases and inhibitors of such kinases.\",\n \"As examples, protein binding assays, nucleic acid binding assays and gel shift assays are useful approaches.\",\n \"Exemplary assays include assaying labeled nuclear receptor (e.g.\",\n \"ER) binding to immobilized MNAR, labeled MNAR or MNAR peptide binding to immobilized nuclear receptor (e.g.\",\n \"ER), etc.\",\n \"Many appropriate assays are amenable to scaled-up, high throughput usage suitable for volume drug screening.\",\n \"The particular assay used will be determined by the particular nature of the MNAR interactions.\",\n \"Assays may employ a single MNAR, MNAR fragments, MNAR fusion products, partial MNAR complexes, or the complete basal transcription complex comprising an MNAR nucleic acid.\",\n \"Useful agents are typically those that bind to or modify the association of MNAR, the MNAR-nuclear receptor complex, or the MNAR-ER complex.\",\n \"Preferred agents include those capable of modulating the expression of MNAR genes, particularly genes transcribed by members of the nuclear receptor superfamily.\",\n \"Useful agents are found within numerous chemical classes, though typically they are organic compounds; preferably small organic compounds.\",\n \"Small organic compounds have a molecular weight of more than 50 yet less than about 2,500, preferably less than about 750, more preferably, less than about 250.Exemplary classes include peptides, saccharides, steroids, and the like.\",\n \"Selected agents may be modified to enhance efficacy, stability, pharmaceutical compatibility, and the like.\",\n \"Structural identification of an agent may be used to identify, generate, or screen additional agents.\",\n \"For example, where peptide agents are identified, they may be modified in a variety of ways to enhance their stability, such a using an unnatural amino acid, such as a D-amino acid, particularly D-alanine, by functionalizing the amino or carboxyl terminus, e.g., for the amino group, acylation or alkylation, and for the carboxyl group, esterification or amidification, or the like.\",\n \"Other methods of stabilization may include encapsulation, for example, in liposomes, etc.\",\n \"Agents may be prepared in a variety of ways known to those skilled in the art.\",\n \"For example, peptides under about 60 amino acids can be readily synthesized today using conventional commercially available automatic synthesizers.\",\n \"Alternatively, peptide (and protein and nucleic acid agents) are readily produced by known recombinant technologies.\",\n \"These assay systems could also be used to screen for potential therapeutics, including peptide and chemical ligands.\",\n \"Such potential therapeutics, activators or inhibitors of MNAR function, could act by either modulating an interaction between MNAR and either a nuclear receptor (e.g.\",\n \"ER) or a kinase (“class I”) or by modulating an activity of MNAR or a nuclear receptor (e.g.\",\n \"ER) or kinase (“class II”).\",\n \"Interaction assays, including but not limited to two-hybrid assays, immunoprecipitation assays, and surface plasmon resonance (SPR) assays, could be used to identify “class I” therapeutics.\",\n \"For example, chemical compounds or peptides can be screened for their ability to modulate the interaction between MNAR and a nuclear receptor (e.g.\",\n \"ER) by SPR.\",\n \"Activity assays, including but not limited to mammalian transfection assays in which nuclear receptor (e.g.\",\n \"ER) transcriptional activity is observed, could be used to identify “class II” compounds.\",\n \"For example, compounds could be screened for their ability to modulate the enhancement of nuclear receptor (e.g.\",\n \"ER) activity by MNAR.\",\n \"Such compounds may modulate the interactions of MNAR with a nuclear receptor (e.g.\",\n \"ER) or they may modulate a known or unknown activity of MNAR or a nuclear receptor (e.g.\",\n \"ER), including a transcriptional activity or assays that can be used to evaluate enzymatic activity of kinases.\",\n \"For therapeutic uses, the compositions and selected agents disclosed herein may be administered by any convenient way that will depend upon the nature of the compound.\",\n \"For small molecular weight agents, oral administration is preferred and enteric coatings may be indicated where the compound is not expected to retain after exposure to the stomach environment.\",\n \"Generally the amount administered will be empirically determined by routine dose-response experiments.\",\n \"Typically amounts range from about 1 to 1000 μg/kg of recipient.\",\n \"Large proteins are preferably administered parenterally, conveniently in a physiologically acceptable carrier, e.g., phosphate buffered saline, saline, deionized water, or the like.\",\n \"Typically, such compositions are added to a retained physiological fluid such as blood or synovial fluid.\",\n \"Other additives may be included, such as stabilizers, bactericides, etc.\",\n \"These additives will be present in conventional amounts.\",\n \"MNAR may also be modified with a label capable of providing a detectable signal either directly or indirectly.\",\n \"Exemplary labels include radioisotopes, fluorescers, etc.\",\n \"Alternatively, a MNAR may be expressed in the presence of a labeled amino acid such as 35S-methionine.\",\n \"Such labeled MNAR and analogs thereof find use, for example, as probes in expression screening assays for proteins that interact with MNAR, or, for example, MNAR binding to ER or other ER interacting proteins or transcription factors in drug screening assays.\",\n \"Specific polyclonal or monoclonal antibodies that can distinguish MNAR from other nuclear proteins are readily made using the methods and compositions disclosed in Harlow and Lane, Antibodies, A Laboratory Manual, Cold Spring Harbor Laboratory, 1988, other references cited herein, as well as immunological and hybridoma technologies known to those in the art.\",\n \"In particular, MNAR and analogs and fragments thereof also find use in raising anti-MNAR antibodies in laboratory animals such as mice and rabbits as well as the production of monoclonal antibodies by cell fusion or transformation.\",\n \"Anti-MNAR antibodies and fragments (Fab, etc.)\",\n \"thereof find use in modulating MNAR role in transcription complexes, screening MNAR expression libraries, etc.\",\n \"In addition, these antibodies can be used to identify, isolate, and purify structural analogs of MNAR.\",\n \"Anti-MNAR antibodies also find use for subcellular localization of MNAR under various conditions such as infection, during various cell cycle phases, induction with cytokines, protein kinases such as C and A, etc.\",\n \"Other exemplary applications include using MNAR-specific antibodies (including monoclonal or MNAR-derived peptide specific antibodies) to immuno-deplete in vitro transcription extracts and using immuno-affinity chromatography to purify MNAR, including analogs, or other nuclear factors which interact with MNAR.\",\n \"Compositions are also provided for therapeutic intervention in disease, for example, by modifying MNARs or MNAR encoding nucleic acids.\",\n \"Oligopeptides can be synthesized in pure form and can find many uses in diagnosis and therapy.\",\n \"These polypeptides can be used, for example, to modulate native MNAR interaction with MNAR interacting proteins.\",\n \"The oligopeptides will generally be more than six and fewer than about 60 amino acids, more usually fewer than about 30 amino acids, although large oligopeptides may be employed.\",\n \"A MNAR or a portion thereof may be used in purified form, generally greater than about 50%, usually greater than about 90% pure.\",\n \"Methods for purifying such peptides to such purities include various forms of chromatographic, chemical, and electrophoretic separations disclosed herein or otherwise known to those skilled in the art.\",\n \"Transgenic Animals and Cells Animal models serve as useful vehicles for screening of compounds, for example, for effects on cholesterol levels, cholesterol absorption, and bile acid synthesis.\",\n \"Transgenic animals, defined as animals with engineered genomes, may prove particularly useful in certain applications.\",\n \"In the present application, the focus is on the modulation of ER activity, or other nuclear receptors and the role of MNAR in effecting the non-genomic activity of ER.\",\n \"Thus, embodiments of this invention include transgenic mice that lack MNAR activity as well as transgenic animals that may lack nuclear receptor activity, ER activity, or a kinase activity.\",\n \"Animals lacking one of the above are useful to determine the specificity of a compound or ligand for MNAR, ER, a kinase.\",\n \"These animals are useful not only in elucidating the biology of this signaling pathway, but as tools in examining possible therapeutic compositions and methods of treatment.\",\n \"For these reasons, transgenic non-human animals of the present invention will have at least one non-functional endogenous MNAR (and/or ER and/or kinase).\",\n \"It further is contemplated that the transgenic non-human animal of the present invention may have both MNAR and ER altered, thus providing cells which lack functional MNAR-ER complex.\",\n \"The non-functional attribute(s) can contain an interruption of the gene coding sequence, a nonsense mutation that truncates the protein product, a deletion of the gene coding sequence or alterations in the MNAR regulatory region.\",\n \"In designing a heterologous gene for expression in animals, sequences which interfere with the efficacy of gene expression, such as coding sequences, promoters, introns, polyadenylation signals, polymerase II termination sequences, hairpins, consensus splice sites and the like, may be eliminated.\",\n \"Current advances in transgenic approaches and techniques have permitted the manipulation of a variety of animal genomes via gene addition, gene deletion, or gene modifications (Franz et al., 1997).\",\n \"The transgenic synthesis of human hemoglobin (U.S. Pat.\",\n \"No.\",\n \"5,602,306, specifically incorporated herein by reference) and fibrinogen (U.S. Pat.\",\n \"No.\",\n \"5,639,940, specifically incorporated herein by reference) in non-human animals have also been disclosed, each specifically incorporated herein by reference in it entirety.\",\n \"The construction of a transgenic mouse model has recently been used to assay potential treatments for Alzheimer's disease (U.S. Pat.\",\n \"No.\",\n \"5,720,936, specifically incorporated herein by reference in its entirety).\",\n \"It is contemplated in the present invention that transgenic animals contribute valuable information as models for studying the effects of ligands specific for the MNAR and/or ER in transgenic MNAR and ER knockout animal models.\",\n \"Conclusions This invention is based on the molecular cloning, structural analysis and characterization of a protein modulator of non-genomic activity of nuclear receptors (MNAR).\",\n \"The present invention demonstrates that human MNAR interacted with nuclear hormone receptors to form a complex and affect the enzymatic activity of Src protein kinases.\",\n \"Crosstalk between cell surface form of ER and intracellular signaling pathways was first demonstrated more than 20 years ago (Pietras, 1975), (Pietras, 1977).\",\n \"Since then multiple reports confirmed rapid estrogen effects occurring in the vasculature, breast, bone, uterus and neuronal tissues that cannot be explained by the “genomic” action of estrogen receptor.\",\n \"It became clear that biological activity of estrogen in responsive cells is mediated not only through a specific high-affinity receptor, located exclusively in their nuclei, but also through the interaction of estrogen with cell-surface receptor.\",\n \"The nature of this receptor is debated.\",\n \"Some evidence supports the existence of a unique membrane receptor, other favors the identity or the strong similarity between the cell-surface and nuclear estrogen receptors (Pietras and Szego, 1979), (Bression et al., 1986), (Pappas et al., 1995).\",\n \"Important evidence, based on experiments with COS cells transfected with ERa cDNA, clearly demonstrated that classical estrogen receptor can activate cSrc in the process leading to activation of tyrosine phosphorylation/p21ras/MAP kinase pathway (Migliaccio et al., 1996).\",\n \"Evaluating the spectrum of ER interacting proteins in MCF7 cells, we discovered a novel ER-interacting protein—MNAR.\",\n \"We showed that both ERα and ERβ interacted with MNAR and affinity of this interaction is enhanced by 17β-estradiol and reduced by 4(OH) tamoxifen (FIG.\",\n \"3).\",\n \"Protein homologous to MNAR—p160 was recently isolated using pull-down approach with SH2 domain of Lck.\",\n \"Our data indicate that MNAR is the major form of this protein expressed in MCF7 cells (FIG.\",\n \"2).\",\n \"Next we evaluated whether MNAR affects estrogen receptors interaction with Src family kinases.\",\n \"Our data indicate that both ERα and ERβ interact with partially purified cSrc and this interaction is not enhanced by 17β-estradiol (FIGS.\",\n \"4A and B).\",\n \"This result does not contradict previously reported data (Migliaccio et al., 2000), (Migliaccio et al., 1998) since we used in vitro transcribed/translated ER and MNAR and also purified cSrc.\",\n \"At the same time Migliaccio et al.\",\n \"(Migliaccio et al., 2000), (Migliaccio et al., 1998) performed their experiment by pulling down Src- or ER-interacting proteins from cell extracts that may contain MNAR.\",\n \"Importantly, in presence of MNAR, estrogen receptors interact with cSrc and Lck in ligand-dependent way (FIGS.\",\n \"4A and B).\",\n \"We also observed that MNAR itself binds to cSrc or Lck with low affinity.\",\n \"Our results therefore suggest that ER and MNAR synergistically bind to Src family kinases.\",\n \"Under basal conditions, the catalytic domain of Src family kinases is constrained in an inactive state through intermolecular interactions.\",\n \"Binding of the SH2 domain to the C-terminal phosphorylated tyrosine locks the molecule in an inhibited conformation ([Matsuda, 1990]).\",\n \"Binding of the SH3 domain to the proline-reach domain of the linker region is also important for kinase inactivation ([Superti-Furga, 1993]).\",\n \"Full catalytic activation requires release of these restrains.\",\n \"The kinase activity of Src can be enhanced by binding of the SH2 domain to phosphotyrosine-containing sequences and by binding of the SH3 domain to the proline-rich sequences ([Hubbard, 1998]).\",\n \"We next investigated if ER-MNAR binding leads to cSrc activation.\",\n \"Our results suggest that MNAR strongly enhances cSrc catalytic activity that can be further stimulated by addition of ER-E2 (FIG.\",\n \"5).\",\n \"Phosphotyrosine 537 of ERα has been identified as the residue essential for ER interaction with SH2 domain of Src ((Migliaccio et al., 1998)).\",\n \"We submit that ER interacts with LXXLL motifs of the MNAR molecule and ER's phosphotyrosine 537 interacts with SH2 domain of Src.\",\n \"At the same time proline-rich domain of MNAR probably interacts with SH3 domain of Src.\",\n \"Formation of this complex leads to cSrc activation.\",\n \"Activation of cSrc in response to EGF has been previously shown to promote the ER phosphorylation by the MAP kinase and stimulation of ER-mediated transcription ([Kato, 1996]; [Bunone, 1996]).\",\n \"Our results indicate that overexpression of MNAR, presumably through activation of cSrc-mediated phosphorylation cascade, augments ER transcriptional activity (FIG.\",\n \"6A).\",\n \"Attenuation of MNAR-induced ER activation in the presence of Src inhibitors PP2 and genistein (FIG.\",\n \"6B) supports the view that MNAR interacts with ER via the cSrc-mediated phosphorylation cascade.\",\n \"Our studies describe a new ER-interacting protein that modulates the cross talk between ER and Src family tyrosine kinases, thus regulating the nongenomic activity of estrogens and potentially other steroids.\",\n \"The nongenomic activity represent an important mechanism by which estrogens regulates cell grows and proliferation.\",\n \"MNAR therefore, may represent an important target for development of new estrogens with more selective action.\",\n \"The above conclusions are supported by the following examples, which are offered by way of illustration and not by way of limitation.\",\n \"EXAMPLES Equipment and Reagents.\",\n \"17β-estradiol (E2) and 4(OH) tamoxifen were obtained from Sigma.\",\n \"ICI-182,780 was provided by AstraZeneca Pharmaceuticals (Wlimington.\",\n \"DE).\",\n \"5′RACE primers were from marathon cDNA kit (BD Biosciences Clontech, Palo Alto, Calif.).\",\n \"Purified cSrc or Lck were from Upstate (Charlottesville, Va.).\",\n \"Biotinylated peptide derived from p34cdc2 was synthesized by the Peptide Chemistry group at Wyeth Research.\",\n \"Glutathione agarose beads were obtained from Sigma-Aldrich (St. Louis, Mo.).\",\n \"Anti-phosphotyrosine antibody and SuperSignal Elisa Pico peroxidase substrate were from Perbio Science AB (Pierce, Rockford, Ill.).\",\n \"Src inhibitor PP2 was from Calbiochem-Novobiochem Corporation (San Diego, Calif.).\",\n \"Method 1: Isolation and identification of MNAR.\",\n \"GST-ERβ-LBD and GST-ERα-LBD were expressed in BL21 (DE3) E. coli (GST is Glutathione S-Transferase).\",\n \"Cell culture in LB at OD0.6 was induced with 0.1 mM IPTG, incubated for 3 hr at 25° C. and cells were pelleted by centrifugation.\",\n \"Bacterial cells were sonicated 5 ml of 20 mM Tris pH 7.2, 500 mM NaCl, 1 mM DTT, 0.2 mM EDTA, 0.1 mM PMSF, centrifuged at 40,000 g for 30 min.\",\n \"To immobilize GST-ERβ-LBD and GST-ERα-LBD cell extract was incubated with 50% glutathione agarose slurry in 20 mM Tris pH 7.2, 180 mM NaCl, 1 mM DTT, 0.005% NP40 (Binding Buffer) at 4° C. MCF7 cell extract was prepared using NE-PER Nuclear and Cytoplasmic Extraction Reagents (Perbio Science AB, Pierce, Rockford, Ill.) according to manufacture instructions.\",\n \"0.5 mg of MCF7 cell extract was incubated at 4° C. with either GST or GST-ER-LBD fusion protein with or without 17-β-estradiol (0.1 μM) at 4° C. Bound material was boiled off the agarose beads in 2× SDS buffer and purified on 10% PAA Gel.\",\n \"The gel was silver stained.\",\n \"Method 2: Mass Spectrometric Analysis of Purified Proteins.\",\n \"Protein samples purified by electrophoresis were excised manually, then reduced, alkylated and digested with trypsin (Promega, Madison, Wis.) in situ using an automated in-gel tryptic digestion robot ((Houthaeve T, 1997)).\",\n \"After digestion, peptide extracts were concentrated to a final volume of 10-20 μl using a Savant Speed Vac Concentrator (ThermoQuest, Holdbrook, N.Y.).\",\n \"Peptide extracts were analyzed on an automated microelectrospray reversed phase HPLC.\",\n \"In brief, the microelectrospray interface consisted of a Picofrit fused silica spray needle, 50 cm length by 75 um ID, 8 um orifice diameter (New Objective, Cambridge Mass.)\",\n \"packed with 10 um C18 reversed-phase beads (YMC, Wilmington, N.C.) to a length of 10 cm.\",\n \"The Picofrit needle was mounted in a fiber optic holder (Melles Griot, Irvine, Calif.) held on a home-built base positioned at the front of the mass spectrometer detector.\",\n \"The rear of the column was plumbed through a titanium union to supply an electrical connection for the electrospray interface.\",\n \"The union was connected with a length of fused silica capillary (FSC) tubing to a FAMOS autosampler (LC-Packings, San Francisco, Calif.) that was connected to an HPLC solvent pump (ABI 140C, Perkin-Elmer, Norwalk, Conn.).\",\n \"The HPLC solvent pump delivered a flow of 50 uL/min which was reduced to 250 nL/min using a PEEK microtight splitting tee (Upchurch Scientific, Oak Harbor, Wash.), and then delivered to the autosampler using an FSC transfer line.\",\n \"The LC pump and auto sampler were each controlled using their internal user programs.\",\n \"Samples were inserted into plastic autosampler vials, sealed, and injected using a 5 ul sample loop.\",\n \"Extracted peptides from the surface digests were concentrated 10-fold using a Savant Speed Vac Concentrator (ThermoQuest, Holdbrook, N.Y.), and then were separated by the microelectrospray HPLC system using a 50 minute gradient of 0-50% solvent B (A: 0.1M HoAc, B: 90% MeCN/0.1M HoAc).\",\n \"Peptide analyses were done on a Finnigan LCQ-DECA ion trap mass spectrometer (ThermoQuest, San Jose, Calif.) operating at a spray voltage of 1.5 kV, and using a heated capillary temperature of 125° C. Data were acquired in automated MS/MS mode using the data acquisition software provided with the instrument.\",\n \"The acquisition method included 1 MS scan (375-600 m/z) followed by MS scans of the top 2 most abundant ions in the MS scan.\",\n \"The instrument then did a second MS scan (600-1000 m/z) followed by MS/MS scans of the top 2 most abundant ions in that scan.\",\n \"The dynamic exclusion and isotope exclusion functions were employed to increase the number of peptide ions that were analyzed (settings: 3 amu=exclusion width, 3 min=exclusion duration, 30 secs=pre-exclusion duration, 3 amu=isotope exclusion width).\",\n \"Automated analysis of MS/MS data was performed using the SEQUEST computer algorithm ((Houthaeve T, 1997)) incorporated into the Finnigan Bioworks data analysis package (ThermoQuest, San Jose, Calif.) using the non-redundant (NR) database of proteins obtained from the NCBI genome center.\",\n \"Method 3: MNAR Cloning: Three pairs of oligonucleotides were designed and used to clone the N-terminal, middle, and C-terminal portions of MNAR Oligo #1: TAGGATCCAGATGGCGGCAGCCGTTCTGAG (SEQ ID NO 3) and oligo #2: CGATCAGGATCCCAAAGC (SEQ ID NO 4) were used to clone the N-terminal portion.\",\n \"Oligo #3: GCTTTGGGATCCTGATCG (SEQ ID NO 5) and oligo #4: CAAGGAGATCTCCACATC (SEQ ID NO 6) were used to clone the middle portion.\",\n \"Oligo #5: GATGTGGAGATCTCCTTG (SEQ ID NO 7) and oligo #6: GCTAGGAGTCAGGCTCTG (SEQ ID NO 8) were used to clone the C-terminal portion.\",\n \"1×107 MCF7 cells were lyzed in Trizol (Life Technologies, Rockville, Md.)\",\n \"for total RNA isolation according to protocol.\",\n \"40 ng of total RNA and 400 nM of oligos was used in a RT-PCR reaction using One Step RT-PCR kit (Qiagen, Valencia, Calif.) to amplify MNAR starting with 30 min at 50° C., followed by 15 min at 95° C., and then 30 cycles of 30 sec at 95° C., 30 sec at 55° C., 60 sec at 72° C., and extension at 72° C. for 7 min.\",\n \"The respective products were cloning into pT-Adv vector (Invitrogen, Carlsbad, Calif.) and sequenced.\",\n \"Full length MNAR was assembled by restriction enzyme digestion and ligation.\",\n \"MNAR was also independently cloned by using oligo #1 and #6 from a human lymphoma marathon cDNA library according to protocol (BD Biosciences Clontech, Palo Alto, Calif.).\",\n \"The MNAR cloned from MCF-7 was identical to the MNAR cloned from marathon cDNA library.\",\n \"A 5′RACE was performed using oligo #7 CCGMGCCAAGACACACAGTGCTGCTGGMTAG (SEQ ID NO 9) and adaptor primer 1 oligo form marathon cDNA kit (BD Biosciences Clontech, Palo Alto, Calif.) to obtain additional sequence information.\",\n \"Method 4: MNAR Expression Analysis.\",\n \"Total RNA (10 ug) from HepG2, SaOS2, HOBs 02/02, COS-7, MCF-7.T47D, MDA231, Ishikawa, 1-10, LNCaPLN3, LNCaPFGC, JCA, TSUPRL cells were prepared by Trizol, separated on gel, transferred to membrane, and probed with radiolabeled oligo #7 corresponding to nucleotide 367 to 400 of cloned MNAR.\",\n \"Another oligo probe #8 CTGGAGAAAAAAGGGGCAGAGATAAAGAGT (SEQ ID NO 10), corresponding to nucleotide 1480 to 1509 of deposited Genbank sequence, was used to probe MCF-7 RNA in another Northern.\",\n \"A second MNAR expression analysis was performed.\",\n \"Radiolabeled oligonucleotide probe corresponding to nucleotide 367 to 393 of MNAR was hybridized to a human multi-tissue northern blot II (BD Biosciences Clontech, Palo Alto, Calif.) in Perfect Hybridization buffer (Sigma-Aldrich, St. Louis, Mo.)\",\n \"at 42° C. The blot was washed three times in 0.2×SSC, 0.1% SDS at 42° C. and exposed to film.\",\n \"β-Actin probe was used as a control.\",\n \"Method 5: Transient Transfection Analysis: HepG2 and COS-7 cells were routinely maintained in DMEM supplemented with 10% FBS.\",\n \"Either 50,000 cells per well of HepG2 or 30,000 per well of COS-7 were seeded in 96 well plates in DMEM without phenol red supplemented with 10% charcoal stripped serum.\",\n \"100 ng of 2×ERE-tk-Luc reporter, 10 ng of pCMV-O-gal internal control, either 1 ng of pcDNA3.1 ERα or 5 ng of pcDNA3.1 ERβ expression vector with or without increasing amount of pcDNA3.1 MNAR or pcDNA3.1 SRC3 expression plasmids were transfected using 0.5 ul of lipofectamine 2000 (Life Technologies, Rockville, Md.)\",\n \"according to protocol.\",\n \"Cells were stimulated either with 10−8M 17-b-estradiol, tamoxifen, or ICI for 24 hr before being processed for luciferase and β-Galactosidase activity.\",\n \"Method 6: MNAR-ER Interaction Analysis: MNAR-Nuclear receptors interaction analysis was performed.\",\n \"SF9 cells extract, overexpressing full-length flag-MNAR, were incubated with in vitro transcribed/translated [35S]-labeled full-length ERα (FIG.\",\n \"5A), ERβ (FIG.\",\n \"5B), androgen (AR) (FIG.\",\n \"5C) and glucocorticoid receptors (GR) (FIG.\",\n \"5D) with and without their corresponding ligands, all at 1 μM.\",\n \"Formed complexes were isolated using anti-FLAG sepharose beads, boiled in 2×SDS buffer and loaded on SDS gel.\",\n \"The gel was dried and subjected to autoradiography.\",\n \"Method 7: ER-MNAR-cSrc Interaction Analysis.\",\n \"In vitro transcribed/translated ERα with (FIG.\",\n \"6A, lanes 4, 5 and 8, 9) or without MNAR (FIG.\",\n \"6A, lanes 2, 3 and 6, 7) in presence (FIG.\",\n \"6A, lanes 3, 5, 7 and 9) or absence of 1 μM E2 (FIG.\",\n \"6A, lanes 2, 4, 6 and 8), were incubated with purified cSrc (FIG.\",\n \"6A, lanes 2-5) or Lck (FIG.\",\n \"6A, lanes 6-9).\",\n \"Formed complexes were isolate by pulled-down with anti-cSrc or anti-Lck antibodies and protein A agarose, boiled in 2×SDS buffer and resolved on an SDS-PAGE gel.\",\n \"Gel was dried and subjected to autoradiography.\",\n \"Method 8: Analysis of cSrc Enzymatic Activity.\",\n \"The cSrc tyrosine kinase activity was evaluated in the absence (FIG.\",\n \"8, lanes 1 and 2) or presence of ER (FIG.\",\n \"8, lanes 3-6), 17β-estradiol (FIG.\",\n \"8, lanes 3 and 5), and MNAR (FIG.\",\n \"8, lanes 2, 5 and 6) using acidified enolase as a substrate.\",\n \"Before reaction proteins were incubated for 1 hr at 4° C. Phosporylation reaction was started by addition of [32P]-γATP and continued at 30° C. for 10 min.\",\n \"Reaction was stopped by addition of 2×SDS buffer.\",\n \"Phosphorylated proteins were separated by SDS-PAGE and the dried gel was subjected to autoradiography.\",\n \"Method 9: MNAR Effect on ER-Mediated Transcription.\",\n \"A—HepG2 cells were transfected with expression plasmids for ERα, MNAR and/or SRC3.Luciferase gene expression, driven by a 2×ERE-tk-reporter, was evaluated in cells treated with E2, 4(OH)-tamoxifen, or ICI 182,780 all at 10 nM.\",\n \"B—HepG2 cells were transfected with expression vectors for ERαand MNAR.\",\n \"Luciferase gene expressions, driven by a 2xERE-tk-reporter, were evaluated in cells treated with 10 nM E2, 10 nM E2 plus 1 μM PP2, or 1 μM PD98059.C—MCF-7 cells were transfected with 100 nM antisense (AS) CATGGAGATGTCCCGGAACAGTGCA (SEQ ID NO: 11) from Sequitur, Inc. (Natick, Mass.)\",\n \"or reverse control (C) oligomers complexed with 1 μg/ml of Lipofectamine 2000 in white media.\",\n \"After 24 hours cells were stimulated with 10 nM E2, 24 hours later cells were lysed in guanidium buffer and total RNA was isolated from lysates.\",\n \"TaqMan analysis was performed using a primer/probe set designed to target MNAR, pS2 and Cathepsin D. D—Extracts of MCF7 cells transfected with 100 nM antisense (AS)—1, or reverse control (C)—2 oligomers stimulated with 10 nM E2 were used for Western Blotting Analysis with rabbit polyclonal MNAR antiserum.\",\n \"MNAR antiserum was generated against 11-mer peptide encoding amino acids 509-520 (SHRKGDSNANSD) (SEQ ID NO: 31) of the MNAR.\",\n \"Method 10: MNAR and ER Interaction and Activation of Src Kinase MNAR and ERα interact with Src and promote phosphorylation of endogenous proteins.\",\n \"MCF-7 cells transfected or not transfected with FLAG-MNAR expression vector were not treated (FIG.\",\n \"10, lanes 1 and 3) or treated with E2 at 10 nM for 5 minutes (lanes 2 and 4).\",\n \"FLAG-MNAR (panel A), cSrc (panel B), or ERα (panels C, D and E) were immunoprecipitated from the cell extracts using corresponding antibodies.\",\n \"Immunoprecipitates were incubated with [32P]γATP for 30 minutes at 30° C. (panels A-C), or with [32P]-γATP and enolase (panel D).\",\n \"Kinase reaction was stopped by boiling in 2×SDS buffer.\",\n \"Phosphorylated material was loaded on SDS gel, which was then dried and subjected to autoradiography.\",\n \"Material obtained by ERα pull down from cells not transfected with FLAG-MNAR, untreated (1) and treated with E2 (2) was probed with MNAR-antiserum (panel E).\",\n \"Method 11: MNAR affect on E2 Induced Erk Kinase 1 and 2 Activation It has been previously reported that activation of Src by estradiol triggers Ras/Erks kinase pathway.\",\n \"We expected that MNAR induced Src activation may lead to activation of Erk kinases 1 and 2.We evaluated the level of Erk activation by E2 in MCF7 cells, cells overexpressing MNAR, and cells in which MNAR expression was substantially attenuated by treatment with MNAR antisense oligonucleotide (SEQ ID NO:11).\",\n \"Antibodies that recognize phosphorylated Erk 1 and 2 were used to detect E2 induced stimulation of Erk 1 and 2 phosphorylation.\",\n \"Extracts of MCF7 cells (FIG.\",\n \"11, lanes 3 and 4) transfected with MNAR expression plasmid (FIG.\",\n \"11, lanes 1 and 2), or MNAR antisense oligonucleotide at 100 nM (FIG.\",\n \"11, lanes 5 and 6); unstimulated (FIG.\",\n \"11, lanes 1, 3 and 5), or stimulated for 5 minutes (FIG.\",\n \"11, lanes 2, 4 and 6) with 10 nM E2 were used for Western blotting analysis with antibodies against phosphorylated Erk 1 and 2—panel A; Erk1/2 antibodies—panel B; MNAR antibodies—panel C. MNAR antisense oligonucleotide (SEQ ID NO:11) was developed at Sequitur Inc. (Natick, Mass.).\",\n \"Method 12: MNAR Structural-Functional Analysis MNAR sequence analysis revealed that the N-terminal portion of its molecule contains multiple LXXLL motifs previously found to be important for interaction with nuclear hormone receptors, while at the C-terminus, MNAR contains a region rich in both proline and glutamic acid.\",\n \"To delineate the regions of MNAR that are responsible for interaction with estrogen receptor and c-Src tyrosine kinase, we performed functional evaluation of MNAR deletion mutants (FIG.\",\n \"12).\",\n \"A series of C-terminal truncations of MNAR was generated using specially designed oligonucleotides, by amplifying appropriate fragments and cloning them into appropriate expression vectors.\",\n \"These mutants were tested for their abilities to enhance ER-stimulated transcription when transfected into HepG2 cells.\",\n \"Method 13: Screening for Compounds that Affect Kinase Activity.\",\n \"Compounds are screened as follows.\",\n \"The reaction buffer used is 50 mM Tris-HCl, pH 7.0, 50 mM MgCl2, 50 uM Na3VO4.The substrate peptide is obtained and 6 mg dissolve in 1.7 mL of RB and store at −20° C. ATP to be used is at 400 uM final concentration.\",\n \"An inhibitor is to be used, for example, Piceatannol—3 mM (stored at 4° C. and protected from light).\",\n \"First, ER incubation with E2.For example, 10 μl of commercially available ER (Panvera) is incubated with 40 μl of RB and 0.5 μl of E2 at 10−6M.\",\n \"The mix is incubated at RT for 1 hour.\",\n \"Next, the mixture is placed in the presence or absence of MNAR, E2, and ER.\",\n \"The solutions are incubated at RT for 30 minutes.\",\n \"To each tube, 10 μl ATP (final concentration 400 μM) is added along with 10 μl substrate peptide.\",\n \"This is incubated for 60 minutes at 30° C. The reaction is stopped by adding 10 μl of 3 mM piceatannol.\",\n \"Now the solution is incubated at RT for 5 minutes.\",\n \"Dilution and peptide immobilization is performed.\",\n \"10 μl Reaction is diluted with 20 ml H2O.\",\n \"200 μl of dilution is added to wells of NeutrAvidin Plate Washing is performed by adding 200 μl of PBS 5× for 3 minutes each.\",\n \"This is followed by addition of Anti-P-Tyr antibody.\",\n \"The antibody is diluted in PBS and BSA and the solution is covered and allowed to incubate at 37° C. for one hour.\",\n \"The mixture is washed the same as above.\",\n \"Then a substrate reaction is generated by adding 100 μl of Pierce substrate and reading in Victor using the HRP protocol.\",\n \"Experimental Analysis and Results Example 1 MNAR Isolation and Identification, Cloning of cDNA Encoding MNAR.\",\n \"To better understand the tissue-selective action of ER ligands, we established a proteomics approach to evaluate the expression and activity of ER-interacting proteins in different cell lines.\",\n \"We identified MNAR using GST-pull down approach.\",\n \"A GST-ERβ ligand-binding domain (LBD) was incubated with a MCF-7 cell extract.\",\n \"FIG.\",\n \"2A presents a silver-stained gel of the fractions obtained by the pull-down experiment.\",\n \"A number of proteins were detected that specifically interacted with GST-ERβ-E2, but not with GST-ERβ, or GST alone.\",\n \"Corresponding bands were excised, digested with trypsin, concentrated, and separated by the microelectrospray HPLC system.\",\n \"Peptide analysis was done on a Finnigan LCQ-DECA ion trap mass spectrometer (ThermoQuest, San Jose, Calif.).\",\n \"Automated analysis of mass spectrometry data was performed using the SEQUEST computer algorithm ([Houthaeve T, 1997]) incorporated into the Finnigan Bioworks data analysis package (ThermoQuest, San Jose, Calif.) using the non-redundant database of proteins obtained from the NCBI genome center.\",\n \"FIG.\",\n \"2A shows a silver-stained gel resulting from the experimental steps as outlined above in Method 1.Cell extract was incubated with Glutathione beads—1, bacterially expressed GST-ERβ-LBD fusion protein in the absence of ligand—2, or presence of E2—3, or 4(OH) tamoxifen—4, both at 10−7M.\",\n \"ER-interacting proteins were isolated using anti-GST agarose and separated on SDS-PAGE.\",\n \"The gel area containing the corresponding band was excised and subjected to trypsin digestion.\",\n \"The bands corresponding to the proteins revealing ligand-dependent interaction were excised, digested with trypsin and identified using mass spectrometry based peptide microsequencing.\",\n \"Together with known and well-characterized coactivators, such as SRC3 and DRIP205, we identified a protein that had not been previously shown to interact with ERs.\",\n \"This protein interacted with the GST-ERβ-LBD in the presence of E2 and had an apparent molecular weight of approximately 120 kDa (FIG.\",\n \"2A).\",\n \"Peptide sequence analysis revealed that this protein could be related to a proline and glutamic acid rich protein—-p160 (Genbank U88153), previously isolated by pull-down with the Src homology domain 2 (SH2) of p56lck (Lck) (21).\",\n \"FIG.\",\n \"2B presents sequences of the peptides identified by the mass spectrometry and their position in the MNAR molecule.\",\n \"Searching the NCI database we also identified several ESTs that matched this protein.\",\n \"One of them, AL03939, matched at the N-terminus and extended the 5′ sequence an additional 100 bp.\",\n \"The aligned sequence was used to design oligonucleotide primers to clone this protein from MCF-7 cells (22).\",\n \"However, the sequence of the cloned protein (see FIG.\",\n \"1) differed substantially from the p160 (Genbank U88153).\",\n \"To differentiate between the two sequences, and to avoid confusion with the family of nuclear receptor coactivators that also are referred as p160s ((Voegel et al., 1998), (Hong et al., 1997), (Anzick et al., 1997), [Suen, 1998]), we named the protein that we cloned from MCF7 cells MNAR.\",\n \"Sequence alignment of MNAR and p160 (FIG.\",\n \"1A) indicated that MNAR does not contain nucleotides from 1075 to 1510 and 3125 to 3151 that are present in the p160 (Genbank U88153).\",\n \"In addition, there were 10 single base pair gaps and one double base pair gap compared to the p160 sequence.\",\n \"MNAR was also independently cloned from a human lymphoma Marathon cDNA library (Invitrogen).\",\n \"Importantly, the MNAR cloned from MCF-7 cells was identical to the MNAR cloned using marathon cDNA library.\",\n \"To obtain additional sequence information, we performed the 5′RACE with primers from marathon cDNA kit (Clonetech).\",\n \"Using 5′RACE, stop codons were found in all three reading frames 5′ of the putative start codon of MNAR.\",\n \"Conceptual translation of the MNAR clone resulted in an 1130 amino acid protein with a calculated molecular weight of 119.6 kDa.\",\n \"This clone contained all peptide sequences initially identified by mass spectrometric analysis (FIG.\",\n \"2B).\",\n \"Sequence analysis has revealed multiple LXXLL motifs localized in the N-terminal portion of MNAR molecule.\",\n \"We putatively call it nuclear receptor interaction domain (NRID).\",\n \"Similar motifs in other transcription factors have been shown to interact with a hydrophobic groove on the surface of the ligand-binding domain of nuclear hormone receptors (Heery et al., 1997), (Torchia et al., 1997) (23).\",\n \"Interesting features of the MNAR molecule are extended proline (PRD) and glutamic acid (ERD) rich domains localized in the C-terminal part of the MNAR molecule.\",\n \"No homology to these domains has been found in the NCl database.\",\n \"Example 2 MNAR Expression Analysis.\",\n \"MNAR expression in HepG2, SaOS2, HOBs 02/02, COS-7, MCF-7.T47D, MDA231, Ishikawa, 1-10, LNCaPLN3, LNCaPFGC, JCA, TSUPRL cell lines was evaluated using Northern blotting analysis.\",\n \"For this purpose we used an oligo-probe derived from the N-terminal portion of MNAR, which is common between MNAR and the deposited p160 sequence (Genbank U88153).\",\n \"Expression of MNAR mRNA was detected in almost all cell lines (FIG.\",\n \"3), as estimated by the presence of a 4 kb band (FIG.\",\n \"3).\",\n \"The level of the MNAR expression varied significantly among different cell lines.\",\n \"It was high in liver (HepG2) and prostate (LNCaPLN3, LNCaPFGC, JCA, TSUPRL) cells, and low to undetectable in breast tumor cells that do not express ER (T47D, MDA231) (FIG.\",\n \"3).\",\n \"To evaluate whether p160 is expressed in MCF-7 cells we used Northern blotting analysis with oligo-probe to the region of p160 that is missing in MNAR sequence and is encoded by nucleotides 1480 to 1509 of deposited to Genbank p160 sequence.\",\n \"Using this probe we failed to detect any message (data not shown).\",\n \"This result indicates that the major form of this protein, expressed in MCF-7 cells, matches well with the MNAR, while the mRNA encoding the p160 is not expressed or expressed at very low levels.\",\n \"MNAR distribution in different tissues was analyzed by Northern blotting.\",\n \"Again, we used an oligo-probe derived from the N-terminal portion of MNAR, which is common between MNAR and the deposited p160 sequence (Genbank U88153).\",\n \"Once again, expression of MNAR mRNA was detected in almost all tissues (FIG.\",\n \"4), as estimated by the presence of a 4 kb band (FIG.\",\n \"4).\",\n \"The level of MNAR expression appears to vary among different tissues.\",\n \"Example 3 MNAR Specifically Interacts with Estrogen Receptors With full-length flag-MNAR, we further evaluated whether MNAR interaction with estrogen receptors is affected by ER ligands.\",\n \"Full-length flag-MNAR, overexpressed in SF9 cells using baculoviral expression system, was used in a pull-down experiment with in vitro transcribed/translated full-length unliganded ER α and β, or receptors liganded with E2, or 4-OH tamoxifen.\",\n \"Formed complexes were isolated using anti-FLAG sepharose beads.\",\n \"FIGS.\",\n \"5A and 5B reveals that ER α and β interact with MNAR, and that this interaction is enhanced by E2 and inhibited by 4(OH) tamoxifen.\",\n \"We also evaluated MNAR interaction with some other nuclear hormone receptors.\",\n \"Full-length flag-MNAR was incubated with transcribed/translated androgen (AR) and glucocorticoid receptors (GR) with and without their corresponding hormone.\",\n \"The data indicated that AR and GR ligand dependently interacted with MNAR (FIGS.\",\n \"5B and 5C).\",\n \"This interaction may explicate the nongenomic activity of these receptor ligands.\",\n \"Detailed investigation of these and other nuclear hormone receptor's crosstalk with MNAR and the Src phosphorylation cascade is an important subject for future investigation.\",\n \"Example 4 MNAR interaction with c-Src and Lck is enhanced by E2-liganded estrogen receptors.\",\n \"Considering that a protein homologous to MNAR, p160, was initially identified via pull-down with the SH2 domain of p56lck (Lck), we next examined whether MNAR interacts with members of Src family tyrosine kinases and affects their interaction with ER.\",\n \"To address this question, we have used in vitro transcribed/translated estrogen receptors ERα, ERβ and MNAR.\",\n \"ER with and without 17β-estradiol (E2), alone or together with MNAR were incubated with commercially available, partially purified cSrc or Lck (Upstate Biotechnology).\",\n \"Formed complexes were pulled-down with anti-cSrc or anti-Lck antibodies and protein A sepharose and then resolved on an SDS gel.\",\n \"The data indicate that ER a interacts with cSrc and Lck (FIG.\",\n \"6A, lanes 2 and 3, 6 and 7).\",\n \"Surprisingly, this interaction was inhibited by E2.When ERa was incubated with both MNAR and cSrc (FIG.\",\n \"6A, lanes 4 and 5) or Lck (FIG.\",\n \"6A, lanes 8 and 9), significant stimulation of Src-ER-MNAR complex formation was detected in the presence of E2 (FIG.\",\n \"6A, lanes 5 and 9) compared to unliganded ERa (FIG.\",\n \"6A, lanes 4 and 8).\",\n \"Since MNAR itself does not interact well with cSrc or Lck (FIG.\",\n \"6A, lane 11), and interaction of cSrc and/or Lck with ERs is inhibited by E2 (FIG.\",\n \"6A, lane 3,), we conclude that all three proteins strongly interact in the presence of E2 and that MNAR is the key to ER-Src interaction.\",\n \"Identical data were obtained for ERβ (FIG.\",\n \"6B).\",\n \"Previous studies from Auricchio and colleagues suggested that ER directly binds to the SH2 domain of Src and that this binding is enhanced by E2.A potential explanation for the apparent inconsistency between our data and theirs may rely on the fact that to evaluate ER-Src interaction they used Src pull-downs from cell extracts.\",\n \"MNAR was most likely present in these extracts and would have mediated ER-Src complex formation.\",\n \"Under basal conditions, the catalytic domain of Src is constrained in an inactive state through intramolecular interactions.\",\n \"Binding of the SH2 domain to the C-terminal phosphorylated tyrosine and SH3 domain to proline-rich region in Src linker domain locks the molecule in an inhibited conformation.\",\n \"(FIG.\",\n \"7).\",\n \"Full catalytic activation requires release of these restraints.\",\n \"The kinase activity of Src can be enhanced by binding of phosphotyrosine-containing sequences to the SH2 domain and binding of proline-rich sequences to the SH3 domain.\",\n \"Considering that MNAR contains an extended proline-rich domain (PRD) and the ligand-binding domain of ER contains a tyrosine residue that can be phosphorylated, we hypothesized that these interactions may lead to Src activation (see FIG.\",\n \"7).\",\n \"Example 5 MNAR Stimulates ER-Driven Transactivation.\",\n \"Activation of cSrc in response to EGF has been previously shown to promote ER phosphorylation and to promote phosphorylation of other transcription factors by MAP kinase, thereby stimulating ER-mediated transcription ([Kato, 1996]; [Bunone, 1996]).\",\n \"To evaluate whether MNAR affects transcriptional activity of ER, COS7 cells were cotransfected with ERα (FIG.\",\n \"6A) or ERβ (FIG.\",\n \"6B) and MNAR.\",\n \"Luciferase gene expression driven by 2×ERE-tk-reporter was evaluated in cells treated and untreated with 17β-estradiol.\",\n \"Our data indicate that increased MNAR expression correlates with stimulation of ERα and ERβ transcriptional activity (FIGS.\",\n \"6A and 6B).\",\n \"Next we evaluated whether cSrc inhibitors affect MNAR-promoted ER activation.\",\n \"HepG2 cells transfected with ERα and MNAR (FIG.\",\n \"8) were treated for 24 hr with 17β-estradiol alone, with 17β-estradiol and selective inhibitor of Src—PP2 (Calbiochem) or with genistein.\",\n \"As in the previous experiment (FIG.\",\n \"8) MNAR stimulated E2 induced ERα activity.\",\n \"At the same time Src inhibitors PP2 and genesteine inhibited MNAR stimulation (FIG.\",\n \"8).\",\n \"These data indicate that MNAR induced stimulation of ER activity is linked to the activation of a phosphorylation cascade.\",\n \"Example 6 MNAR Overexpression affects Transcriptional Activity of ER To evaluate if MNAR overexpression affects transcriptional activity of ER, HepG2 cells were transiently cotransfected with plasmids for expression of ERα and MNAR.\",\n \"Luciferase gene expression controlled by a 2×ERE-tk-reporter was evaluated in cells treated with E2, 4(OH)-tamoxifen, or ICI 182,780 all at 10 nM.\",\n \"The results (FIG.\",\n \"9A) indicate that increasing MNAR expression correlates well with stimulation of ERα transcriptional activity.\",\n \"Similar stimulation of ER transcriptional activity was observed with overexpression of the ER-coactivator, SRC3.Our data also indicate that 4(OH)-tamoxifen and ICI 182,780 do not support MNAR stimulation of ER activity.\",\n \"Similar results were observed with ERβ (data not shown).\",\n \"To investigate if cSrc activation is responsible for the MNAR stimulation of ER activity, HepG2 cells transfected with ERα, MNAR and 2×ERE-tk-reporter plasmid (FIG.\",\n \"9B) were treated with E2 alone, or E2 and either the cSrc inhibitor, PP2, or the MEK kinase inhibitor, PD98059.We found that both PP2 and PD98059 abrogate MNAR stimulation of ER activity.\",\n \"These results indicate that MNAR promoted stimulation of ER transcriptional activity is linked to the activation of the MAP kinase phosphorylation cascade.\",\n \"We hypothesize that ER phosphorylation by Erk, downstream from Src and MEK kinases, may be responsible for enhancement of ER transcriptional activity, as previously described.\",\n \"In addition, it is possible the that activation of the Src/MAP kinase pathway leads to phosphorylation of some other transcription factors, that are important for ER transcriptional activity.\",\n \"Importantly, these data also suggest that the so-called “nongenomic” action of nuclear hormone receptors through activation of the phosporylation cascade may regulate activity of transcription factors, and, by doing so, ultimately influence gene expression.\",\n \"We speculate that this mechanism may institute the crosstalk between nuclear receptors that are able to interact with MNAR and other transcription factors whose activity is regulated by phosphorylation.\",\n \"Example 7 MNAR Affects ER-Mediated Gene Expression We next used an antisense approach to assess MNAR's role in estrogen regulation of gene transcription.\",\n \"Antisense oligonucleotides (SEQ ID NO:11) were developed (Sequitur Inc., Natick, Mass.)\",\n \"that significantly inhibited MNAR expression in MCF-7 cells, as estimated by TaqMan (FIG.\",\n \"9C) and Western Blotting (FIG.\",\n \"9D) analyses.\",\n \"MCF-7 cells were transfected with 100 nM antisense or reverse control oligomers.\",\n \"After 24 hours, cells were stimulated with 10 nM E2, and 24 hours later cells were lysed and total RNA was isolated.\",\n \"TaqMan analysis was performed using a primer/probe set designed to target MNAR, pS2 and Cathepsin D—two genes known to be regulated by estrogens in MCF-7 cells.\",\n \"FIG.\",\n \"9C presents the levels of MNAR, pS2 and Cathepsin D mRNA expression, normalized to GAPDH mRNA.\",\n \"Interestingly expression of MNAR mRNA was substantially stimulated by E2 treatment.\",\n \"Our data indicate that reduction in MNAR expression lead to dramatic reduction of E2-stimulated pS2 and Cathepsin D expression.\",\n \"Importantly, the MNAR antisense oligonucleotides did not significantly affect the basal level of these genes expression.\",\n \"We expect that ER phosphorylation is probably important for E2-mediated expression of pS2 and Cathepsin D. These data support our position that MNAR is essential for ER regulation of gene expression.\",\n \"Example 8 MNAR and ER Interaction and Activation of Src Kinase in MCF-7 Cells We evaluated whether MNAR and ER interacted and activated Src kinase in MCF-7 cells.\",\n \"MCF-7 cells transfected with a Flag-MNAR expression vector were treated with E2 at 10 nM for 5 minutes.\",\n \"Flag-MNAR (FIG.\",\n \"10, panel A), ERα (FIG.\",\n \"10, panel C and D), and csrc (FIG.\",\n \"10, panel B) were immunoprecipitated from the cell extracts using their corresponding antibodies.\",\n \"Material precipitated using ERα antiserum was probed with rabbit polyclonal anti-MNAR antibody (FIG.\",\n \"10, panel E).\",\n \"E2-enhancement of ERα-MNAR interaction was clearly detected.\",\n \"Considering that MCF7 cells not transfected with FLAG-MNAR were used for this experiment, this data confirm that endogenous MNAR and ERα interact in MCF7 cells.\",\n \"Immunoprecipitates were incubated with [32P]γATP at 30° C. for 30 minutes.\",\n \"Reaction was stopped by boiling in 2×SDS buffer and resolved on a SDS PMG.\",\n \"Strong MNAR- and E2-dependent phosphorylation of several endogenous proteins was detected.\",\n \"Especially evident was phosphorylation of an endogenous protein with an apparent molecular weight of approximately 34 kDa that coprecipitated with cSrc, ER, and MNAR (see FIG.\",\n \"10).\",\n \"Importantly phosphorylation of this protein was detected in material coprecipitated with anti-Src and anti-ER antibodies from cells untransfected with MNAR (FIG.\",\n \"10, lanes 1 and 2).\",\n \"The phosphorylation was stimulated in presence of E2 (FIG.\",\n \"10, lane 2) and then dramatically augmented in cells transfected with MNAR (FIG.\",\n \"10, lanes 3 and 4).\",\n \"These data strongly support our position that MNAR, ER, and cSrc interact in MCF-7 cells, and that the MNAR-ER complex strongly stimulates cSrc kinase activity, promoting phosphorylation of some endogenous protein in MCF-7 cells.\",\n \"An identical phosphorylation pattern was detected when enolase was used as Src substrate (FIG.\",\n \"10, panel D).\",\n \"Example 9 MNAR Affects E2 Induced Erk Kinase 1 and 2 Activation We next evaluated the level of Erk activation by E2 in MCF7 cells (see FIG.\",\n \"11, lanes 3 and 4) cells overexpressing MNAR (FIG.\",\n \"11, lanes 1 and 2) and cells in which MNAR expression was substantially attenuated by treatment with MNAR antisense oligonucleotide (FIG.\",\n \"11, lanes 5 and 6).\",\n \"Strong E2 induced stimulation of Erk 1 and 2 phosphorylation was clearly detected using antibodies that recognize phosphorylated Erk 1 and 2 (FIG.\",\n \"11, panel A).\",\n \"This activation was significantly enhanced in cells overexpressing MNAR (FIG.\",\n \"11, panel A, lanes 1 and 2) and attenuated in cells treated with MNAR antisense oligonucleotides (FIG.\",\n \"11, panel A, lanes 5 and 6).\",\n \"Neither treatment with MNAR antisense oligonucleotides nor short treatment with E2 affected the level of total Erk 1 and 2 (FIG.\",\n \"11, panel B).\",\n \"Western blotting analysis indicated that MNAR level was strongly augmented in cells transfected with MNAR expression plasmid (FIG.\",\n \"11, panel C, lanes 1 and 2) and attenuated in cells treated with MNAR antisense oligonucleotides (FIG.\",\n \"11, panel C, lanes 5 and 6).\",\n \"These data indicate that MNAR controls E2 induced stimulation of cSrc and Erk 1 and 2 kinases.\",\n \"Example 10 MNAR Structural-Functional Organization MNAR sequence analysis revealed that the N-terminal portion of its molecule contains multiple LXXLL motifs found to be important for interaction with nuclear hormone receptors.\",\n \"At the C-terminus, MNAR contains a region rich in both proline and glutamic acid.\",\n \"To delineate the regions on MNAR responsible for interaction with estrogen receptors and c-src tyrosine kinase, a functional evaluation of MNAR deletion mutants was performed (FIG.\",\n \"12).\",\n \"A series of C-terminal truncations of MNAR was generated using specially designed oligonucleotides, by amplifying appropriate fragments and cloning them into appropriate expression vectors.\",\n \"These mutants were tested for their abilities to enhance ER-stimulated transcription when transfected into HepG2 cells.\",\n \"While MNAR mutants containing aa 1-469 (mutant 2, SEQ ID NO: 14), 1-278 (mutant 3, SEQ ID NO:15), and 1-189 (mutant 4, SEQ ID NO:16) are still able to augment ER transcriptional activity.\",\n \"Mutants containing the C-terminal portion of MNAR molecule aa 595-1131 (mutant 9, SEQ ID NO: 17), 595-962 (mutant 10, SEQ ID NO: 18), and 595-887 (mutant 11, SEQ ID NO: 19)) were inactive.\",\n \"These data suggest that the domain that is necessary and sufficient to stimulate ER transcriptional activity is localized to the N-terminal part of NRID between aa 1 and 189 (FIG.\",\n \"13).\",\n \"This region contains the LXXLL motifs—4th (aa154-159) (SEQ ID NO: 20), 5th (aa 176-181) (SEQ ID NO: 21), and 6th (aa 181-185) (SEQ ID NO: 22) (FIG.\",\n \"12).\",\n \"To evaluate which of these motifs is responsible for interacting with estrogen receptor, we generated mutants of MNAR with these three LXXLL motifs mutated individually or in combination.\",\n \"While MNAR utilizes the LXXLL motifs to interact with estrogen receptors, MNAR uses PXXP motifs to interact with Src.\",\n \"MNAR contains three of these PXXP motifs within the N-terminal portion.\",\n \"From the deletion analysis, constructs of MNAR missing the first two PXXP motifs are not able to stimulate estrogen receptor activity in transfection assays (FIG.\",\n \"13).\",\n \"This suggests these first two PXXP motifs are sufficient for interaction with Src.\",\n \"Mutation of these motifs revealed that the first PXXP motif (aa 55-58) (PXXP mutant 1, SEQ ID NO: 23) is necessary and sufficient to activate ER transcriptional activity, while the second PXXP motif (aa 64-67) (PXXP mutant 2 SEQ ID NO: 32) is dispensable (FIG.\",\n \"14).\",\n \"Additional mutants generated in this study were as follows: mutant 5 with aa 1-120 (SEQ ID NO: 24); mutant 6 with aa 1-79 (SEQ ID NO: 25); mutant 7 with aa 1-40 (SEQ ID NO: 26); mutant 8 with aa 190-469 (SEQ ID NO: 27); mutant 12 with aa 888-1131 (SEQ ID NO: 28); mutant 13 with aa 80-594 (SEQ ID NO: 29); and mutant 14 with aa 80-574 (SEQ ID NO: 30).\",\n \"Each of these mutants, along with the other mutants listed above, were schematically diagrammed in FIG.\",\n \"12 and were analyzed for their ability to enhance ER-stimulated transcription when transfected into HepG2 cells.\",\n \"(FIG.\",\n \"13) In conclusion, we demonstrate that interaction of estrogen receptors α and β with MNAR is enhanced by E2 but not by 4(OH) tamoxifen.\",\n \"We also show that the MNAR-ER complex interacts with members of Src family of tyrosine kinases—p60src (Src) and p56lck (Lck) dramatically stimulating Src enzymatic activity.\",\n \"We also show that MNAR, through activation of the Src phosphorylation cascade, affects ER transcriptional activity and ultimately ER-mediated gene expression.\",\n \"Specifically, we demonstrate that short term treatment of MCF-7 cells with E2 leads to MNAR-Src-ER complex formation, activation of Src and phosphorylation of some endogenous proteins.\",\n \"Enhanced MNAR expression in transiently transfected cells leads to stimulation of ER transcriptional activity.\",\n \"Since specific inhibitors of Src and MEK kinases block the ability of MNAR to augment ER transcriptional activity, we conclude that this activation of ER is due to stimulation of the MAP kinase phosphorylation cascade promoted by activation of Src kinases.\",\n \"Alternatively, depletion of MNAR from MCF-7 cells using antisense oligonucleotides inhibits ER-mediated gene expression, supporting its involvement in ER-regulated gene expression.\",\n \"These data reveal that MNAR modulates ER crosstalk with Src family tyrosine kinases and by doing so mediates the 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(1994).\",\n \"Signal transduction by lymphocyte antigen receptors.\",\n \"Cell 76, 263-74.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473753"}}},{"rowIdx":748,"cells":{"text":{"kind":"list like","value":[["Mobile sleeve structure for maintaining spatial relationship between vertebrae","An arrangement and method of use for supporting a plurality of vertebrae in a cervical column in predetermined spatial relation and for facilitating a spinal fusion procedure.","A plurality of templates (12, 14) couple to respectively associated vertebrae at anterior lateral surfaces thereof.","Such coupling is effected with fasterners, such as bone screws (19).","The templates are slidingly coupled to each other by a coupler assembly (22) whereby they are displaceable along a path over a limited distance that is substantially parallel to the longitudinal axis of the cervical column.","The sliding displacement between the templates is limited by a pair of protruding tongues (27) that extend through elongated apertures (29).","The elongated dimension of the apertures determines the extent of sliding travel, which extent is established to accommodate subsidence of the bone fusion and to prevent separation of the sliding elements of the coupler assembly.","The coupler assembly is coupled to each template by engagement between posts (18) and post holes (15).","Additionally, locking plates and fasteners, such as threaded fasteners, are used to ensure the security of the engagement between the templates and the coupler assembly.","Sequential replication of the arrangement enables three or more vertebra to be supported in the determined spatial relation."],["1.An implantable mobile sleeve for stabilizing the anterior side of a vertebral column, comprising: a lowermost template having respective anterior and posterior surfaces, and a through-hole penetrating through the anterior and posterior surfaces, wherein the through-hole is configured to receive a fastener, said lowermost template having a width dimension that is greater than a height dimension; an uppermost template having respective anterior and posterior surfaces, and a through-hole penetrating through the anterior and posterior surfaces, wherein the through-hole is configured to receive a fastener, said uppermost template having a width dimension that is greater than a height dimension; a coupling assembly for coupling said lowermost and uppermost templates adjustably to one another, said coupling assembly extending substantially vertically and orthogonal with respect to the width dimensions of said lowermost and uppermost templates, said coupling assembly having; a bottom sleeve portion having a first end for coupling to said lowermost template and a second end extending upward toward said uppermost template, and a top sleeve portion having a first end with at least one post aperture for coupling to said uppermost template, and a second end that extends toward said lowermost template and configured to terminate at an opening that forms a hollow longitudinal channel that extends between the first and second ends of the top sleeve portion, the second end of the top sleeve portion being configured to engage slidably with the second end of the bottom sleeve portion, and a locking plate for securing a selectable one of the top and bottom sleeve portions to its corresponding template.","2.The implantable mobile sleeve structure of claim 1 wherein the bottom sleeve of said coupling assembly has a greater thickness at the second end thereof than at its first end.","3.The implantable mobile sleeve structure of claim 1 wherein the lowermost template is arranged for coupling to the anterior region of a first cervical vertebra and the uppermost template is arranged for coupling to the anterior region of a second cervical vertebra.","4.The implantable mobile sleeve structure of claim 3 wherein the uppermost and lowermost templates are each formed in response to the curvatures of corresponding surfaces of the first and second vertebrae.","5.The implantable mobile sleeve structure of claim 1, wherein said top sleeve portion is configured to have a channel for accommodating slidingly the second end of said bottom sleeve portion.","6.The implantable mobile sleeve structure of claim 5 further comprising: a tongue extending orthogonally from the second end of said bottom sleeve portion, and an elongated aperture disposed in the second portion of said top sleeve portion, said elongated aperture accommodating therein said tongue, whereby a displacement limit is imposed on the slidable engagement between the bottom and top sleeve portions.","7.The implantable mobile sleeve structure of claim 6, wherein the elongated aperture is disposed in a base portion of the channel.","8.The implantable mobile sleeve structure of claim 1, wherein said uppermost and lowermost templates communicate exclusively on an anterior substantially vertical portion of their corresponding vertebrae.","9.An implantable mobile sleeve for stabilizing the anterior side of a vertebral column, comprising: a lowermost template having a generally rectangular configuration having an elongated width dimension perpendicular to the axis of a vertebral column, said lowermost template having an anterior surface, a posterior surface for communicating with exclusively with a lateral anterior surface of a vertebra, and an outer edge extending between the anterior surface and the posterior surface, there being further provided at least two pre-drilled through-holes penetrating through the lowermost template between the anterior surface to the posterior surface; an uppermost template having a generally rectangular configuration having an elongated width dimension perpendicular to the axis of a vertebral column, the uppermost template having an anterior surface, a posterior surface that will rest against a second vertebra and, an outer edge between extending between the anterior surface and the posterior surface, there being further provided two or more pre-drilled through-holes penetrating through the uppermost template between the anterior surface to the posterior surface; at least two posts attached to the anterior surface of the uppermost template and the lowermost template, and arranged to extend orthogonally outward from the anterior surface; a plurality of fasteners disposed through the at least two pre-drilled holes of the uppermost template and the lowermost template, corresponding ones of said plurality of fasteners attaching the lowermost template to the first vertebra and the uppermost template to the second vertebra, wherein the lengthwise side of the lowermost template and the lengthwise side of the uppermost template are parallel when the lowermost template is attached to the first vertebra and the uppermost template is attached to the second vertebra, and a generally rectangular shaped adjustable sleeve having a longitudinal dimension thereof extending parallel to a longitudinal axis of the vertebral column, the adjustable sleeve having a bottom sleeve portion having a first end with at least two pre-drilled post holes for coupling the bottom sleeve portion to the posts located on the lowermost template and a second end that extends upward toward a top sleeve portion of the adjustable sleeve having a first end with two or more pre-drilled post holes that couple the top sleeve portion to the posts located on the uppermost template, wherein the top sleeve portion extends downward toward the lowermost template and terminates at a second end that forms an opening to a hollow longitudinal channel that extends between the first end and second end of the top sleeve portion, the second end and the hollow sleeve being configured to receive slidably the second end of the bottom sleeve portion; a sliding displacement limiter arrangement located on the anterior surface of the top sleeve portion of the adjustable sleeve and having a tongue portion attached to the anterior surface of the bottom sleeve portion that is positioned in the hollow channel of the top sleeve portion, wherein the tongue extends through and slidably connects to an elongated aperture disposed through the anterior surface of the top sleeve portion, and first and second locking plates each associated with one of said uppermost and lowermost templates, said first and second locking plates each having at least two pre-drilled post holes for engaging with the posts of the lowermost template and the uppermost template and thereby securing the adjustable sleeve to the uppermost template and the lowermost template.","10.The implantable mobile sleeve structure of claim 9 further comprising: a second bottom sleeve portion of a second adjustable sleeve placed onto the posts of the uppermost template over the top sleeve portion and under said first locking plate; a second uppermost template having a generally rectangular shape with the lengthwise side of the rectangle extending perpendicular to the axis of a vertebral column, the second uppermost template having an anterior surface, a posterior surface that communicates with a third vertebra, and an outer edge between extending between the anterior surface and the posterior surface, and at least two pre-drilled through-holes penetrating through the second uppermost template between the anterior surface to the posterior surface, wherein the pre-drilled holes are configured to receive two or more fasteners; two posts attached to the anterior surface of the second uppermost template and arranged to extend orthogonally outward therefrom; two fasteners respectively disposed through the two pre-drilled holes of the second uppermost template, each fastener attaching the second uppermost template to the third vertebra, wherein the lengthwise side of the second lowermost template and the lengthwise side of the second uppermost template are parallel when the lowermost template is attached the uppermost template and the second uppermost template is attached to the third vertebra, and a second top sleeve portion of a second adjustable sleeve installed on the posts.","11.The implantable mobile sleeve structure of claim 10 wherein the vertebra are part of an anterior cervical vertebral column.","12.A method of attaching a mobile sleeve between two adjacent vertebra, the method comprising the steps of: attaching a lowermost template to the anterior vertebral body of a first vertebra of a vertebral column using fasteners disposed through pre-drilled holes in the lowermost template; attaching an uppermost template to the anterior vertebral body of a second vertebra of the vertebral column located above and adjacent to the first vertebra using fasteners disposed through pre-drilled holes in the uppermost template; bending a bottom sleeve portion of an adjustable sleeve to conform to the curvature of the vertebral column; placing a first end of the bottom sleeve portion of the adjustable sleeve using pre-drilled post holes located at the first end of the bottom sleeve portion onto a set of posts that extend outward from the anterior surface of the lowermost template; placing a first end of a top sleeve portion of the adjustable sleeve using pre-drilled post holes located at the first end of the top sleeve portion onto posts that extend out of the anterior surface of the uppermost template, and attaching a locking plate onto the posts of the lowermost template and the uppermost template over the adjustable sleeve and the lowermost template and the uppermost template.","13.The method of claim 12 further comprising the further steps of: removing the locking plate located on top of the uppermost template; attaching a second uppermost template of a second mobile sleeve to the anterior vertebral body of a third vertebra located above and adjacent to the second vertebra using fasteners disposed through pre-drilled holes in the second uppermost template; bending a second bottom sleeve portion of a second adjustable sleeve to conform to the curvature of the vertebral column; placing a first end of the second bottom sleeve portion of the second adjustable sleeve using pre-drilled post holes located at the first end the second bottom sleeve portion onto a set of posts that extend outward from the anterior surface of the top plate of the first mobile sleeve; placing a first end of a second top sleeve portion of the second adjustable sleeve using pre-drilled post holes located at the first end the second top sleeve portion onto posts that extend out of the anterior surface of the second uppermost template; attaching a locking plate onto the posts of the uppermost template over the adjustable sleeve and the second adjustable sleeve, and attaching a locking plate onto the posts of the second uppermost template over the adjustable sleeve and the second uppermost template.","14.The method of claim 13 wherein the vertebral column is the an anterior cervical column."],["<SOH> BACKGROUND OF THE INVENTION <EOH>1.Field of the Invention This invention relates generally to spinal implant systems, and more particularly, to a mobile sleeve structure that maintains a determinable spatial relationship between adjacent vertebrae.","2.Description of the Related Art There is a need in the field of neurosurgery for spinal implant systems that enable surgeons to maintain a determinable spatial relationship between adjacent vertebrae during vertebral fusion procedures.","A typical vertebral fusion procedure involves implanting a bone graft or cage element into the intervertebral region known that normally would be occupied by a disc.","Patients that suffer from degenerative disc disorders may be required to undergo a surgical procedure that involves the fusing together ofthe adjacent vertebrae that are located immediately above and below the disc region.","During a vertebral fusion procedure, bone grafts are inserted into the disc region to facilitate fusion of the vertebrae by promoting growth of bone in the disc.","The result is that the vertebrae will eventually fuse together.","Such patients may additionally require a series of such bone grafts that will result in the fusing together of several vertebrae in the cervical vertebral column.","A problem with vertebral fusion procedures is that once the bone grafts are implanted they are prone to becoming misplace due to movement of the adjacent vertebrae.","In response to this problem several systems have been developed that will immobilize the adjacent vertebrae by attaching a plate between the adjacent vertebrae.","The attached plate will maintain the spatial relationship between the vertebrae and promote fusion by preventing slippage of the bone grafts during the fusion process.","One known system for maintaining spinal elements in a desired spatial relationship incorporates a series of plates having predetermined heights and shapes.","Each such plate has a plurality of apertures disposed at lengthwise distal ends of the plate.","Bone screws are threaded through the apertures to attach the plate to a corresponding one of a pair of anterior cervical vertebrae.","The plates so attached maintain a spatial relationship between the two adjacent vertebrae that are located immediately above and below a bone graft implant.","In the known system, the plates are attached to extend longitudinally with respect to the axis of an anterior cervical vertebral column.","It is a deficiency of the above system that plates of various heights must be prefabricated and made available during the procedure because the distance between respective pairs of vertebrae will vary along the length of the anterior cervical column.","Thus, upon the conclusion of the procedure there will remain several unused plates that will have to be disposed of or sterilized.","Another problem with the known system is that the plates are only used between two vertebrae and cannot be used to maintain a desired spatial relationship between three or more vertebrae.","Three or more vertebrae may need to be secured in predetermined relation to one another when the patient is diagnosed to require more than one bone graft implant.","Some systems have been developed that use a one-piece rod that can be attached across several vertebrae.","A problem with the known rod system is that it does not accommodate the natural lordotic curvature of an individual patient's spinal column as the lordotic curvature varies from patient to patient."],["<SOH> SUMMARY OF THE INVENTION <EOH>The foregoing and other objects are achieved by this invention which provides an implantable mobile sleeve for stabilizing the anterior side of a vertebral column.","The implantable mobile sleeve has a group ofgenerally rectangular components that generally have an “T” shape.","The mobile sleeve has a lowermost template with a generally rectangular shape, in that the lowermost template has two distal length dimensions that are longer than the dimensions of two opposing sides that constitute the width of the plate.","When mounted to a vertebra the lengthwise sides of the rectangular shaped lowermost template will extend in a direction substantially perpendicular to the axis of the vertebral column.","The lowermost template has a posterior surface that rests against the surface of a vertebra, and an opposing anterior surface that faces outward from the surface of the vertebra.","The lowermost template has two or more pre-drilled apertures that penetrate the lowermost template from the posterior surface to the anterior surface.","The pre-drilled apertures are configured to receive a fastener to attach the lowermost template to the vertebra.","Two posts extend substantially orthogonally outward from the anterior surface.","The posts facilitate the fastening of other components of the mobile sleeve to the lowermost template.","In some embodiments, the posts are threaded so that a fastener can be used to fix components that are installed on the posts.","In one embodiment of the invention, the mobile sleeve is provided with an uppermost template having a generally rectangular shape.","The uppermost template is substantially identical to the lowermost template, except that the uppermost template is mounted to a second vertebra located above the first vertebra where the bottom plate is mounted.","During attachment, the mobile sleeve is attached between adjacent vertebrae on the vertebral column.","The lowermost template will be coupled to a first vertebra and the uppermost template will be coupled to a second vertebra disposed above the first vertebra.","In a preferred embodiment, the templates are secured to the anterior region of a cervical vertebral column.","However, the mobile sleeve structure described herein may additionally be attached to other regions of the cervical vertebral column, or to other areas of the spine.","Both the uppermost template and the lowermost templates are each affixed to vertebrae using two or more fasteners disposed through the two or more pre-drilled through holes located on the top and lowermost templates.","The fasteners, in this embodiment, are be inserted into the vertebrae at respective angles in order to prevent the templates from being pulled away from the vertebrae.","The fasteners attach the lowermost template to the first vertebra and the uppermost template to the second vertebra.","Such fasteners can, in certain embodiments of the invention, be threaded bone screws, pins, or any other suitable fastener for attaching the templates to the vertebrae.","While only two bone screws are described herein as being used in the practice of the illustrative embodiment of the invention, persons of skill in the art can, in light of the teaching herein, employ a different number of fasteners depending on conditions encountered during the procedure, such as vertebral size or bone density, spatial positioning of the vertebrae, and the strength of attachment required by the physician.","In other embodiments, one or both of the top and lowermost templates are affixed to the vertebrae using adhesives that are applied over the posterior surface of the templates.","Once the bottom plate and the uppermost template are attached to the respective first and second vertebrae, it is preferred that the lengthwise sides of the bottom and uppermost templates be parallel.","In a further embodiment of the invention, a generally rectangular adjustable sleeve slidably connects the uppermost template and the lowermost template by means of the aforementioned posts and/or apertures through, the templates.","It is an advantage of the invention that perfect placement of the adjustable sleeve is achieved because the posts extend generally perpendicular with respect to the midportion of the anterior vertebral body.","The adjustable sleeve has a bottom sleeve portion that perpendicularly couples at a first end to the middle region of the lowermost template.","A second end of the bottom sleeve portion extends upward toward the uppermost template.","With respect to the distance between the anterior and posterior surfaces, the bottom sleeve portion will have a thickness at the first end that differs from that of its second end.","In some embodiments of the invention, the thickness at the first end is about 1 mm and will increase to about 2 mm in the middle region and at second end of the bottom sleeve portion.","The reason for the varying thickness is so that several adjustable sleeves can be stacked onto the posts of the lowermost template without any large protrusions in the lowermost template area.","The thickness of the lowermost template is not limited to 1 mm and 2 mm since sleeves of various thicknesses will be needed depending on the needs of a particular surgical procedure.","In a further embodiment, the adjustable sleeve has a top sleeve portion with a first end that couples to post holes in the middle region of the uppermost template.","The top sleeve portion also extends downward toward the bottom sleeve portion and terminates at a second end.","With regard to thickness (i.e., the distance between the anterior and posterior surfaces), the top sleeve portion will have a different thickness at the first end than at the second end of the top sleeve portion.","Illustratively, the thickness at the first end can be about 1 mm and will increase to about 2 mm as for the middle and second end of the top sleeve portion.","The reason for the varying thickness is so that several adjustable sleeves can be stacked onto the posts of the uppermost template without any large protrusions in the uppermost template area.","The thickness of the uppermost template is not limited to 1 mm and 2 mm since sleeves of various thicknesses will be needed depending on the needs of a particular surgical procedure.","The second end of the top sleeve portion also has an opening to a hollow channel that extends upward toward the first end of the top sleeve portion.","The second end of the bottom sleeve portion enters the bottom end of the top sleeve portion and is slidably disposed through the hollow channel.","The hollow channel affords mobility to the top third of the adjustable sleeve and allows the height of the mobile sleeve to adjust for post-operative subsidence of the vertebrae.","The movement of the adjustable sleeve during subsidence enables the resulting load to be shared equally by all of the components of the mobile sleeve.","Such distribution of the load increases the integrity of the mobile sleeve by preventing the damage to the templates, adjustable sleeve, or the vertebrae that may occur when the load is not equally distributed.","In order to control sliding movement between the top and lowermost templates, a tongue and groove mechanism can, in certain embodiments, be incorporated along the anterior surface of the adjustable sleeve.","One or more tongues protrude from the anterior surface of the bottom sleeve portion that is disposed through the hollow channel, extending orthogonally outward from the anterior surface.","One or more grooves are located on the anterior surface of the top sleeve portion and are arranged to extend completely through the anterior side of the top sleeve portion, whereby the tongues will extend into and slide freely within the grooves.","In the practice of this embodiment, the grooves are of sufficient length to allow a top sleeve portion and a bottom sleeve portion to slide relative to each other and thereby accommodate subsidence of the vertebrae.","However, the grooves will not be so long as to permit the bottom sleeve portion to become separated from the top sleeve portion.","The lowermost two thirds portion of the adjustable sleeve (i.e., the lower portion of the bottom sleeve portion) is formed of a solid material that can be bent with a plate bender so that the adjustable sleeve will correspond substantially to the curvature of the vertebral column.","Although the adjustable sleeve is described herein as having a top third portion that can be adjusted for length and a bottom two thirds that are solid, different ratios are achievable depending upon the region to which the mobile sleeve is to be attached, or the particular needs of a patient.","In a further embodiment, a locking plate holds the adjustable sleeve and template together to allow the distribution of stress to all the parts of the mobile sleeve.","Controlling the distribution of stress contributes to advantageous distribution of the load by ensuring that no one particular portion of the mobile sleeve is burdened excessively by the stress that is caused by the subsidence of the vertebrae.","The locking plate in this embodiment has two or more post holes that are aligned and placed onto the two or more posts of the bottom or uppermost templates.","In order to hold the locking plate in place, the post holes of the locking plate are configured to be secured onto the posts as the locking plate moves into place.","In other embodiments, the posts of the uppermost template and the lowermost template are threaded so that a fastener, such as a nut, can be used to lock the locking plate in place.","In a yet further embodiment of the invention, additional mobile sleeve structures are linked to form additional levels of structure to the spinal support system.","The post holes can be used as a way of linking together mobile sleeve structures when a patient needs more than one implant.","A second mobile sleeve is linked to a first mobile sleeve by removing the locking plate of either the uppermost template or the lowermost template, depending on whether the site of the next bone graft implant is above or below the uppermost template or the lowermost template of the first mobile sleeve.","A second adjustable sleeve is then placed onto the post holes over the first adjustable sleeve and the locking plate is then placed back on to the holes.","A second uppermost template or lowermost template (i. e., depending on whether the second sleeve is being place above or below the first mobile sleeve) is then attached to a vertebra so that the second end of the second adjustable sleeve is placed onto the posts of the second uppermost template or the second lowermost template.","A locking plate then is placed over either the second uppermost template or the second lowermost template and the second end of the adjustable sleeve.","While the second mobile sleeve has been described as attaching so that the mobile sleeves are attached across a series of three adjacent vertebra, the mobile sleeve structure may be attached to three vertebrae that span across more than three vertebrae of the vertebral column.","In addition, more levels may be attached to the mobile sleeve structure to create a device that has any multiple number (i.e., two, three, four, etc.)","of mobile sleeves in the structure.","In accordance with a further aspect of the present invention, there is provided a method of attaching a mobile sleeve between two adjacent cervical vertebrae.","The method includes the steps of: attaching a lowermost template to the anterior vertebral body of a first vertebra using fasteners disposed through pre-drilled holes in the lowermost template; attaching an uppermost template to the anterior vertebral body of a second vertebra located above and adjacent to the first vertebra; and installing fasteners through pre-drilled holes in the uppermost template.","In a further embodiment of this method aspect of the invention, the first and second vertebrae are separated by one or more vertebrae, and therefore are not adjacent to one another.","For instance, the uppermost template and the lowermost template can be attached to respective vertebra in a manner wherein the mobile sleeve spans across any number of vertebrae.","In a further embodiment, there is provided the step of bending the bottom sleeve portion of an adjustable sleeve in response to the curvature of the vertebral column.","Additionally, there are provided the steps of installing the first end of the bottom sleeve portion of the adjustable sleeve onto posts that extend outward of the anterior surface of the lowermost template, and further installing the first end of a top sleeve portion of the adjustable sleeve is placed onto posts that extend out of the anterior surface of the uppermost template.","As previously described, each adjustable sleeve fits onto the posts using pre-drilled post holes located at the first ends of the bottom and top sleeve portions, respectively.","In a further embodiment, there is provided the step of coupling the adjustable sleeve to the top and lowermost templates at a generally perpendicular angle.","In a further embodiment of this method aspect of the invention, there is provided the step of installing a locking plate onto the posts of the uppermost template and the lowermost template over the adjustable sleeve and the uppermost template and lowermost template.","Additionally, there is provided the step of locking the locking plate with a fastener.","Illustratively, when a second level (i.e., a second mobile sleeve) is added to a vertebra located above the second vertebra, there is provided a step of removing the locking plate located on the uppermost template.","Then, there is performed a step of attaching the second uppermost template of the second mobile sleeve to the anterior vertebral body of a third vertebra located above and adjacent to the second vertebra using fasteners disposed through pre-drilled holes in the uppermost template.","In a highly advantageous embodiment of the method aspect of the invention, there is provided the step of bending the bottom sleeve portion of a second adjustable sleeve in response to the curvature of the vertebral column.","A first end of the bottom sleeve portion of the second adjustable sleeve is placed onto posts that extend out of the anterior surface of the uppermost template of the first mobile sleeve.","The first end of a top sleeve portion of the adjustable sleeve is placed onto posts that extend out of the anterior surface of the second uppermost template attached to the third vertebra.","The second adjustable sleeve is configured to fit onto the posts using, pre-drilled post holes located at the first ends of the bottom sleeve portion and top sleeve portion.","The adjustable sleeve couples the top and lowermost templates at a generally perpendicular angle.","There is additionally provided the step of placing locking plates onto the posts of the uppermost template of the first mobile sleeve and the second uppermost template.","In this embodiment, the locking plates are placed over the ends of the second adjustable sleeve and uppermost template of the first mobile sleeve and the second uppermost template.","Then, in this embodiment, there is provided the step of affixing the locking plate with a fastener.","It is to be understood that once the two mobile sleeve devices have been connected, additional levels can be added to this structure by simply repeating the steps recited for attaching the second mobile sleeve.","The mobile sleeves may be coupled in a manner that permits the adjustable sleeve portion to span across more than one vertebrae, as described above.","This provides the significant advantage that the interconnection of several levels of mobile sleeves will constitute a support structure that will be custom fitted to the lordotic curvature of the anterior cervical column."],["RELATIONSHIP TO OTHER PATENT APPLICATION This application claims the benefit of U.S.","Provisional Patent Application No.","60/381,579, filed Apr.","4, 2001.BACKGROUND OF THE INVENTION 1.Field of the Invention This invention relates generally to spinal implant systems, and more particularly, to a mobile sleeve structure that maintains a determinable spatial relationship between adjacent vertebrae.","2.Description of the Related Art There is a need in the field of neurosurgery for spinal implant systems that enable surgeons to maintain a determinable spatial relationship between adjacent vertebrae during vertebral fusion procedures.","A typical vertebral fusion procedure involves implanting a bone graft or cage element into the intervertebral region known that normally would be occupied by a disc.","Patients that suffer from degenerative disc disorders may be required to undergo a surgical procedure that involves the fusing together ofthe adjacent vertebrae that are located immediately above and below the disc region.","During a vertebral fusion procedure, bone grafts are inserted into the disc region to facilitate fusion of the vertebrae by promoting growth of bone in the disc.","The result is that the vertebrae will eventually fuse together.","Such patients may additionally require a series of such bone grafts that will result in the fusing together of several vertebrae in the cervical vertebral column.","A problem with vertebral fusion procedures is that once the bone grafts are implanted they are prone to becoming misplace due to movement of the adjacent vertebrae.","In response to this problem several systems have been developed that will immobilize the adjacent vertebrae by attaching a plate between the adjacent vertebrae.","The attached plate will maintain the spatial relationship between the vertebrae and promote fusion by preventing slippage of the bone grafts during the fusion process.","One known system for maintaining spinal elements in a desired spatial relationship incorporates a series of plates having predetermined heights and shapes.","Each such plate has a plurality of apertures disposed at lengthwise distal ends of the plate.","Bone screws are threaded through the apertures to attach the plate to a corresponding one of a pair of anterior cervical vertebrae.","The plates so attached maintain a spatial relationship between the two adjacent vertebrae that are located immediately above and below a bone graft implant.","In the known system, the plates are attached to extend longitudinally with respect to the axis of an anterior cervical vertebral column.","It is a deficiency of the above system that plates of various heights must be prefabricated and made available during the procedure because the distance between respective pairs of vertebrae will vary along the length of the anterior cervical column.","Thus, upon the conclusion of the procedure there will remain several unused plates that will have to be disposed of or sterilized.","Another problem with the known system is that the plates are only used between two vertebrae and cannot be used to maintain a desired spatial relationship between three or more vertebrae.","Three or more vertebrae may need to be secured in predetermined relation to one another when the patient is diagnosed to require more than one bone graft implant.","Some systems have been developed that use a one-piece rod that can be attached across several vertebrae.","A problem with the known rod system is that it does not accommodate the natural lordotic curvature of an individual patient's spinal column as the lordotic curvature varies from patient to patient.","SUMMARY OF THE INVENTION The foregoing and other objects are achieved by this invention which provides an implantable mobile sleeve for stabilizing the anterior side of a vertebral column.","The implantable mobile sleeve has a group ofgenerally rectangular components that generally have an “T” shape.","The mobile sleeve has a lowermost template with a generally rectangular shape, in that the lowermost template has two distal length dimensions that are longer than the dimensions of two opposing sides that constitute the width of the plate.","When mounted to a vertebra the lengthwise sides of the rectangular shaped lowermost template will extend in a direction substantially perpendicular to the axis of the vertebral column.","The lowermost template has a posterior surface that rests against the surface of a vertebra, and an opposing anterior surface that faces outward from the surface of the vertebra.","The lowermost template has two or more pre-drilled apertures that penetrate the lowermost template from the posterior surface to the anterior surface.","The pre-drilled apertures are configured to receive a fastener to attach the lowermost template to the vertebra.","Two posts extend substantially orthogonally outward from the anterior surface.","The posts facilitate the fastening of other components of the mobile sleeve to the lowermost template.","In some embodiments, the posts are threaded so that a fastener can be used to fix components that are installed on the posts.","In one embodiment of the invention, the mobile sleeve is provided with an uppermost template having a generally rectangular shape.","The uppermost template is substantially identical to the lowermost template, except that the uppermost template is mounted to a second vertebra located above the first vertebra where the bottom plate is mounted.","During attachment, the mobile sleeve is attached between adjacent vertebrae on the vertebral column.","The lowermost template will be coupled to a first vertebra and the uppermost template will be coupled to a second vertebra disposed above the first vertebra.","In a preferred embodiment, the templates are secured to the anterior region of a cervical vertebral column.","However, the mobile sleeve structure described herein may additionally be attached to other regions of the cervical vertebral column, or to other areas of the spine.","Both the uppermost template and the lowermost templates are each affixed to vertebrae using two or more fasteners disposed through the two or more pre-drilled through holes located on the top and lowermost templates.","The fasteners, in this embodiment, are be inserted into the vertebrae at respective angles in order to prevent the templates from being pulled away from the vertebrae.","The fasteners attach the lowermost template to the first vertebra and the uppermost template to the second vertebra.","Such fasteners can, in certain embodiments of the invention, be threaded bone screws, pins, or any other suitable fastener for attaching the templates to the vertebrae.","While only two bone screws are described herein as being used in the practice of the illustrative embodiment of the invention, persons of skill in the art can, in light of the teaching herein, employ a different number of fasteners depending on conditions encountered during the procedure, such as vertebral size or bone density, spatial positioning of the vertebrae, and the strength of attachment required by the physician.","In other embodiments, one or both of the top and lowermost templates are affixed to the vertebrae using adhesives that are applied over the posterior surface of the templates.","Once the bottom plate and the uppermost template are attached to the respective first and second vertebrae, it is preferred that the lengthwise sides of the bottom and uppermost templates be parallel.","In a further embodiment of the invention, a generally rectangular adjustable sleeve slidably connects the uppermost template and the lowermost template by means of the aforementioned posts and/or apertures through, the templates.","It is an advantage of the invention that perfect placement of the adjustable sleeve is achieved because the posts extend generally perpendicular with respect to the midportion of the anterior vertebral body.","The adjustable sleeve has a bottom sleeve portion that perpendicularly couples at a first end to the middle region of the lowermost template.","A second end of the bottom sleeve portion extends upward toward the uppermost template.","With respect to the distance between the anterior and posterior surfaces, the bottom sleeve portion will have a thickness at the first end that differs from that of its second end.","In some embodiments of the invention, the thickness at the first end is about 1 mm and will increase to about 2 mm in the middle region and at second end of the bottom sleeve portion.","The reason for the varying thickness is so that several adjustable sleeves can be stacked onto the posts of the lowermost template without any large protrusions in the lowermost template area.","The thickness of the lowermost template is not limited to 1 mm and 2 mm since sleeves of various thicknesses will be needed depending on the needs of a particular surgical procedure.","In a further embodiment, the adjustable sleeve has a top sleeve portion with a first end that couples to post holes in the middle region of the uppermost template.","The top sleeve portion also extends downward toward the bottom sleeve portion and terminates at a second end.","With regard to thickness (i.e., the distance between the anterior and posterior surfaces), the top sleeve portion will have a different thickness at the first end than at the second end of the top sleeve portion.","Illustratively, the thickness at the first end can be about 1 mm and will increase to about 2 mm as for the middle and second end of the top sleeve portion.","The reason for the varying thickness is so that several adjustable sleeves can be stacked onto the posts of the uppermost template without any large protrusions in the uppermost template area.","The thickness of the uppermost template is not limited to 1 mm and 2 mm since sleeves of various thicknesses will be needed depending on the needs of a particular surgical procedure.","The second end of the top sleeve portion also has an opening to a hollow channel that extends upward toward the first end of the top sleeve portion.","The second end of the bottom sleeve portion enters the bottom end of the top sleeve portion and is slidably disposed through the hollow channel.","The hollow channel affords mobility to the top third of the adjustable sleeve and allows the height of the mobile sleeve to adjust for post-operative subsidence of the vertebrae.","The movement of the adjustable sleeve during subsidence enables the resulting load to be shared equally by all of the components of the mobile sleeve.","Such distribution of the load increases the integrity of the mobile sleeve by preventing the damage to the templates, adjustable sleeve, or the vertebrae that may occur when the load is not equally distributed.","In order to control sliding movement between the top and lowermost templates, a tongue and groove mechanism can, in certain embodiments, be incorporated along the anterior surface of the adjustable sleeve.","One or more tongues protrude from the anterior surface of the bottom sleeve portion that is disposed through the hollow channel, extending orthogonally outward from the anterior surface.","One or more grooves are located on the anterior surface of the top sleeve portion and are arranged to extend completely through the anterior side of the top sleeve portion, whereby the tongues will extend into and slide freely within the grooves.","In the practice of this embodiment, the grooves are of sufficient length to allow a top sleeve portion and a bottom sleeve portion to slide relative to each other and thereby accommodate subsidence of the vertebrae.","However, the grooves will not be so long as to permit the bottom sleeve portion to become separated from the top sleeve portion.","The lowermost two thirds portion of the adjustable sleeve (i.e., the lower portion of the bottom sleeve portion) is formed of a solid material that can be bent with a plate bender so that the adjustable sleeve will correspond substantially to the curvature of the vertebral column.","Although the adjustable sleeve is described herein as having a top third portion that can be adjusted for length and a bottom two thirds that are solid, different ratios are achievable depending upon the region to which the mobile sleeve is to be attached, or the particular needs of a patient.","In a further embodiment, a locking plate holds the adjustable sleeve and template together to allow the distribution of stress to all the parts of the mobile sleeve.","Controlling the distribution of stress contributes to advantageous distribution of the load by ensuring that no one particular portion of the mobile sleeve is burdened excessively by the stress that is caused by the subsidence of the vertebrae.","The locking plate in this embodiment has two or more post holes that are aligned and placed onto the two or more posts of the bottom or uppermost templates.","In order to hold the locking plate in place, the post holes of the locking plate are configured to be secured onto the posts as the locking plate moves into place.","In other embodiments, the posts of the uppermost template and the lowermost template are threaded so that a fastener, such as a nut, can be used to lock the locking plate in place.","In a yet further embodiment of the invention, additional mobile sleeve structures are linked to form additional levels of structure to the spinal support system.","The post holes can be used as a way of linking together mobile sleeve structures when a patient needs more than one implant.","A second mobile sleeve is linked to a first mobile sleeve by removing the locking plate of either the uppermost template or the lowermost template, depending on whether the site of the next bone graft implant is above or below the uppermost template or the lowermost template of the first mobile sleeve.","A second adjustable sleeve is then placed onto the post holes over the first adjustable sleeve and the locking plate is then placed back on to the holes.","A second uppermost template or lowermost template (i. e., depending on whether the second sleeve is being place above or below the first mobile sleeve) is then attached to a vertebra so that the second end of the second adjustable sleeve is placed onto the posts of the second uppermost template or the second lowermost template.","A locking plate then is placed over either the second uppermost template or the second lowermost template and the second end of the adjustable sleeve.","While the second mobile sleeve has been described as attaching so that the mobile sleeves are attached across a series of three adjacent vertebra, the mobile sleeve structure may be attached to three vertebrae that span across more than three vertebrae of the vertebral column.","In addition, more levels may be attached to the mobile sleeve structure to create a device that has any multiple number (i.e., two, three, four, etc.)","of mobile sleeves in the structure.","In accordance with a further aspect of the present invention, there is provided a method of attaching a mobile sleeve between two adjacent cervical vertebrae.","The method includes the steps of: attaching a lowermost template to the anterior vertebral body of a first vertebra using fasteners disposed through pre-drilled holes in the lowermost template; attaching an uppermost template to the anterior vertebral body of a second vertebra located above and adjacent to the first vertebra; and installing fasteners through pre-drilled holes in the uppermost template.","In a further embodiment of this method aspect of the invention, the first and second vertebrae are separated by one or more vertebrae, and therefore are not adjacent to one another.","For instance, the uppermost template and the lowermost template can be attached to respective vertebra in a manner wherein the mobile sleeve spans across any number of vertebrae.","In a further embodiment, there is provided the step of bending the bottom sleeve portion of an adjustable sleeve in response to the curvature of the vertebral column.","Additionally, there are provided the steps of installing the first end of the bottom sleeve portion of the adjustable sleeve onto posts that extend outward of the anterior surface of the lowermost template, and further installing the first end of a top sleeve portion of the adjustable sleeve is placed onto posts that extend out of the anterior surface of the uppermost template.","As previously described, each adjustable sleeve fits onto the posts using pre-drilled post holes located at the first ends of the bottom and top sleeve portions, respectively.","In a further embodiment, there is provided the step of coupling the adjustable sleeve to the top and lowermost templates at a generally perpendicular angle.","In a further embodiment of this method aspect of the invention, there is provided the step of installing a locking plate onto the posts of the uppermost template and the lowermost template over the adjustable sleeve and the uppermost template and lowermost template.","Additionally, there is provided the step of locking the locking plate with a fastener.","Illustratively, when a second level (i.e., a second mobile sleeve) is added to a vertebra located above the second vertebra, there is provided a step of removing the locking plate located on the uppermost template.","Then, there is performed a step of attaching the second uppermost template of the second mobile sleeve to the anterior vertebral body of a third vertebra located above and adjacent to the second vertebra using fasteners disposed through pre-drilled holes in the uppermost template.","In a highly advantageous embodiment of the method aspect of the invention, there is provided the step of bending the bottom sleeve portion of a second adjustable sleeve in response to the curvature of the vertebral column.","A first end of the bottom sleeve portion of the second adjustable sleeve is placed onto posts that extend out of the anterior surface of the uppermost template of the first mobile sleeve.","The first end of a top sleeve portion of the adjustable sleeve is placed onto posts that extend out of the anterior surface of the second uppermost template attached to the third vertebra.","The second adjustable sleeve is configured to fit onto the posts using, pre-drilled post holes located at the first ends of the bottom sleeve portion and top sleeve portion.","The adjustable sleeve couples the top and lowermost templates at a generally perpendicular angle.","There is additionally provided the step of placing locking plates onto the posts of the uppermost template of the first mobile sleeve and the second uppermost template.","In this embodiment, the locking plates are placed over the ends of the second adjustable sleeve and uppermost template of the first mobile sleeve and the second uppermost template.","Then, in this embodiment, there is provided the step of affixing the locking plate with a fastener.","It is to be understood that once the two mobile sleeve devices have been connected, additional levels can be added to this structure by simply repeating the steps recited for attaching the second mobile sleeve.","The mobile sleeves may be coupled in a manner that permits the adjustable sleeve portion to span across more than one vertebrae, as described above.","This provides the significant advantage that the interconnection of several levels of mobile sleeves will constitute a support structure that will be custom fitted to the lordotic curvature of the anterior cervical column.","BRIEF DESCRIPTION OF THE DRAWING Comprehension of the invention is facilitated by reading the following detailed description, in conjunction with the annexed drawings, in which: FIG.","1a is an isometric representation of a single level embodiment of mobile sleeve structure constructed in accordance with the invention; FIG.","1b is a cross-sectional representation taken along line a-a in FIG.","1a; FIG.","2 is a plan representation of a portion of the cervical column with the mobile sleeve structure mounted between two cervical vertebrae; FIG.","3 is a plan representation of the multi-level embodiment of two mobile sleeve structures attached across three cervical vertebrae, and FIG.","4 is a side view of the multi-level embodiment of two mobile sleeve structures attached across three cervical vertebrae.","DETAILED DESCRIPTION FIG.","1a is an isometric representation of the mobile sleeve 10 that is provided with a series of components that when assembled together cause mobile sleeve 10 to have a shape generally in the form of an uppercase letter “I.” A lowermost template 12 and an uppermost template 14 each have an anterior side 16, a posterior side 17 (not shown in this figure), and an outer edge 20 that extends around the perimeter between posterior side 17 and anterior side 16.A respective pair of posts 18 extend orthogonally outward from each of the anterior sides of the lowermost and uppermost templates.","Two posts 18 are located on each anterior side 16 of lowermost template 12 and uppermost template 14.Posts 18 are used to connect lowermost template 12 and uppermost template together and, as will be described hereinafter, to facilitate incorporation of additional levels to the overall structure whereby more than two vertebra are to be fused.","Lowermost template 12 and uppermost template 14 each have two or more pre-drilled through-holes 13 that extend from the posterior side to anterior side of each template.","The through-holes, in this specific illustrative embodiment of the invention, are threaded to receive a threaded fastener 19, which may be a conventional bone screw.","Lowermost template 12 and uppermost template 14 are each also provided with two or more post holes 15 that are configured to receive posts when two or more mobile sleeve structures are linked together as will be described in greater detail in FIG.","3.Each of lowermost template 12 and uppermost template 14 has a generally rectangular shape with a width that is longer than the height.","When installed, the lowermost and uppermost templates are arranged to be substantially parallel.","In the specific illustrative embodiment of the invention, it is preferred that lowermost template 12 and uppermost template 14 are curved as shown so that the posterior sides of both templates are concave.","The curved posterior side of lowermost template 12 and uppermost template 14 ensures that both templates are substantially flush with the curved surface of the anterior cervical vertebrae (not shown) when affixed thereto as herein described.","Conversely, the anterior sides of the lowermost and uppermost templates are convex, and bulky protrusions at the implant site are thereby minimized.","Lowermost template 12 and uppermost template 14 are coupled to one another by a generally rectangular shaped adjustable sleeve 22 that extends substantially vertically in the figure and is coupled substantially orthogonally to the lowermost and uppermost templates at its distal ends.","As shown, adjustable sleeve 22 is formed of a top sleeve portion 24 and a bottom sleeve portion 26 arranged longitudinally distal, and each such portion has respective pre-drilled post holes.","Bottom sleeve portion 26 is connected to lowermost template 12 by registering the posts located on the anterior surface of lowermost template 12 through the post holes.","Similarly, top sleeve portion 24 is connected to uppermost template 14 by registering the posts located on the anterior surface of the uppermost template 14 through the corresponding post holes.","Top sleeve portion 24 and bottom sleeve portion 26 each have a generally rectangular shape and may be slidably connected to their respectively associated templates.","As shown in FIG.","1b, which is cross-sectional representation taken at section line a-a of FIG.","la shows that top sleeve portion 24 has an internal region 28 that has a rectangular shape and extends longitudinally.","Top sleeve portion 24 slides into the end of, and along, hollow channel 28 which cross-sectionally surrounds same.","In order to control the extent of the sliding extension between uppermost template 14 and lowermost template 12 there is provided in this specific illustrative embodiment of the invention, a pair of tongues 27 and corresponding elongated apertures 29 that accommodate the tongues as shown.","Tongues 27 are attached to the anterior surface ofthe bottom sleeve portion 26 portion that is disposed through hollow channel 28.Tongues 27 will extend substantially orthogonal outward from the anterior surface.","The two elongated apertures 29 are located on the anterior surface of top sleeve portion 24.Elongated apertures 29 extend completely through the anterior side of top sleeve portion 24 so that tongues 27 will extend therethrough and slide freely with respect thereto.","The elongated apertures are of sufficient length along their respective major axes to allow top sleeve portion 24 and bottom sleeve portion 26 to be longitudinally movable relative to each other to accommodate subsidence of the vertebrae.","In this specific illustrative embodiment of the invention, a locking plate 30 is installed on the posts of the uppermost template.","Locking plate 30 serves to retain adjustable sleeve 22 and uppermost template 14 in fixed relation to each other.","The locking plate has two pre-drilled post holes 15 that enable the installation of locking plate 30 on the two posts of uppermost template 14.In some embodiments of the invention, a locking plate 30 similar to that described hereinabove is affixed to lowermost template 12.In order to hold the locking plate on the post holes, a known grasping arrangement, such as a resilient locking element (not shown), or a threaded fastened (not shown) can 25 be employed.","FIG.","2 is a plan representation of a portion of the cervical column with mobile sleeve 10 mountedbetweentwo cervical vertebrae 34 and 36 (represented schematically).","Elements of structure that correspond to those discussed in connection with FIGS.","1a and 1b are similarly designated.","The cervical portion of a human spinal column (not shown) has a bone graft 32 placed between first vertebra 36, located below bone graft 32, and a second vertebra 36, located above bone graft 32.Mobile sleeve 10 is coupled to first vertebra 36 and extends upward longitudinally with respect to cervical column to the region where mobile sleeve 10 attaches to second vertebra 36.Mobile sleeve 10 holds first vertebra 34 and second vertebra 36 in a desired spatial relationship relative to each other.","Maintaining the spatial relationship of first vertebra 34 and second vertebra 36 will increase the likelihood of success of spinal fusion procedures by preventing bone graft 32 from shifting and becoming dislodged.","Lowermost template 12 is shown to be coupled to first vertebra 36 by a pair of bone screws 38 that are disposed though pre-drilled through-holes in lowermost template 12 discussed above in connection with FIG.","la.","Uppermost template 14 is shown to be attached to second vertebra 36 with a similar pair of bone screws 38 that are disposed though corresponding pre-drilled through-holes in uppermost template 14.FIGS.","3 and 4 are a plan representation and a side partially cross-sectional representation, respectively, of a multi-level embodiment of two mobile sleeve structures attached across three cervical vertebrae.","Elements of structure that correspond to those in FIGS.","1a, 1b, and 2 are similarly designated.","The cervical portion of a human spinal column having first vertebra 34, second vertebra 36, and third vertebra 40 is shown to have two bone grafts 32, 32′.","Bone graft 32 is dispose between first vertebra 34 and second vertebra 36, and second bone graft 32′ is disposed between second vertebra 36 and third vertebra 40.Mobile sleeve 10 is attached between first vertebra 34 and second vertebra 36 as described in FIG.","2.In this specific illustrative embodiment of the invention, a second level in the form of a second mobile sleeve 10′ is attached between second vertebra 36 and third vertebra 40.A lower sleeve portion 26′ of second mobile sleeve 10′ is aligned and connected to posts 18 of uppermost template 14 of first mobile sleeve 10.Referring to FIG.","4 a locking plate 30 is attached to post 18 over bottom sleeve 26′ and uppermost template 14 just as described in FIG.","2.Uppermost template 14′ of second mobile sleeve 10 is attached with bone screws 38 to third vertebra 40 located above second bone graft 32′.","A top sleeve portion 24′ 30 of second mobile sleeve 10′ is placed on to posts 18 of uppermost template.","Uppermost template 14′ of second mobile sleeve 10′ is can be used, in other embodiments of the invention, for placement of yet a third mobile sleeve (not shown), if needed in the future.","Although the invention has been described in terms of specific embodiments and applications, persons skilled in the art can, in light of this teaching, generate additional embodiments without exceeding the scope or departing from the spirit of the invention described herein.","Accordingly, it is to be understood that the drawing and description in this disclosure are proffered to facilitate comprehension of the invention, and should not be construed to limit the scope thereof."]],"string":"[\n [\n \"Mobile sleeve structure for maintaining spatial relationship between vertebrae\",\n \"An arrangement and method of use for supporting a plurality of vertebrae in a cervical column in predetermined spatial relation and for facilitating a spinal fusion procedure.\",\n \"A plurality of templates (12, 14) couple to respectively associated vertebrae at anterior lateral surfaces thereof.\",\n \"Such coupling is effected with fasterners, such as bone screws (19).\",\n \"The templates are slidingly coupled to each other by a coupler assembly (22) whereby they are displaceable along a path over a limited distance that is substantially parallel to the longitudinal axis of the cervical column.\",\n \"The sliding displacement between the templates is limited by a pair of protruding tongues (27) that extend through elongated apertures (29).\",\n \"The elongated dimension of the apertures determines the extent of sliding travel, which extent is established to accommodate subsidence of the bone fusion and to prevent separation of the sliding elements of the coupler assembly.\",\n \"The coupler assembly is coupled to each template by engagement between posts (18) and post holes (15).\",\n \"Additionally, locking plates and fasteners, such as threaded fasteners, are used to ensure the security of the engagement between the templates and the coupler assembly.\",\n \"Sequential replication of the arrangement enables three or more vertebra to be supported in the determined spatial relation.\"\n ],\n [\n \"1.An implantable mobile sleeve for stabilizing the anterior side of a vertebral column, comprising: a lowermost template having respective anterior and posterior surfaces, and a through-hole penetrating through the anterior and posterior surfaces, wherein the through-hole is configured to receive a fastener, said lowermost template having a width dimension that is greater than a height dimension; an uppermost template having respective anterior and posterior surfaces, and a through-hole penetrating through the anterior and posterior surfaces, wherein the through-hole is configured to receive a fastener, said uppermost template having a width dimension that is greater than a height dimension; a coupling assembly for coupling said lowermost and uppermost templates adjustably to one another, said coupling assembly extending substantially vertically and orthogonal with respect to the width dimensions of said lowermost and uppermost templates, said coupling assembly having; a bottom sleeve portion having a first end for coupling to said lowermost template and a second end extending upward toward said uppermost template, and a top sleeve portion having a first end with at least one post aperture for coupling to said uppermost template, and a second end that extends toward said lowermost template and configured to terminate at an opening that forms a hollow longitudinal channel that extends between the first and second ends of the top sleeve portion, the second end of the top sleeve portion being configured to engage slidably with the second end of the bottom sleeve portion, and a locking plate for securing a selectable one of the top and bottom sleeve portions to its corresponding template.\",\n \"2.The implantable mobile sleeve structure of claim 1 wherein the bottom sleeve of said coupling assembly has a greater thickness at the second end thereof than at its first end.\",\n \"3.The implantable mobile sleeve structure of claim 1 wherein the lowermost template is arranged for coupling to the anterior region of a first cervical vertebra and the uppermost template is arranged for coupling to the anterior region of a second cervical vertebra.\",\n \"4.The implantable mobile sleeve structure of claim 3 wherein the uppermost and lowermost templates are each formed in response to the curvatures of corresponding surfaces of the first and second vertebrae.\",\n \"5.The implantable mobile sleeve structure of claim 1, wherein said top sleeve portion is configured to have a channel for accommodating slidingly the second end of said bottom sleeve portion.\",\n \"6.The implantable mobile sleeve structure of claim 5 further comprising: a tongue extending orthogonally from the second end of said bottom sleeve portion, and an elongated aperture disposed in the second portion of said top sleeve portion, said elongated aperture accommodating therein said tongue, whereby a displacement limit is imposed on the slidable engagement between the bottom and top sleeve portions.\",\n \"7.The implantable mobile sleeve structure of claim 6, wherein the elongated aperture is disposed in a base portion of the channel.\",\n \"8.The implantable mobile sleeve structure of claim 1, wherein said uppermost and lowermost templates communicate exclusively on an anterior substantially vertical portion of their corresponding vertebrae.\",\n \"9.An implantable mobile sleeve for stabilizing the anterior side of a vertebral column, comprising: a lowermost template having a generally rectangular configuration having an elongated width dimension perpendicular to the axis of a vertebral column, said lowermost template having an anterior surface, a posterior surface for communicating with exclusively with a lateral anterior surface of a vertebra, and an outer edge extending between the anterior surface and the posterior surface, there being further provided at least two pre-drilled through-holes penetrating through the lowermost template between the anterior surface to the posterior surface; an uppermost template having a generally rectangular configuration having an elongated width dimension perpendicular to the axis of a vertebral column, the uppermost template having an anterior surface, a posterior surface that will rest against a second vertebra and, an outer edge between extending between the anterior surface and the posterior surface, there being further provided two or more pre-drilled through-holes penetrating through the uppermost template between the anterior surface to the posterior surface; at least two posts attached to the anterior surface of the uppermost template and the lowermost template, and arranged to extend orthogonally outward from the anterior surface; a plurality of fasteners disposed through the at least two pre-drilled holes of the uppermost template and the lowermost template, corresponding ones of said plurality of fasteners attaching the lowermost template to the first vertebra and the uppermost template to the second vertebra, wherein the lengthwise side of the lowermost template and the lengthwise side of the uppermost template are parallel when the lowermost template is attached to the first vertebra and the uppermost template is attached to the second vertebra, and a generally rectangular shaped adjustable sleeve having a longitudinal dimension thereof extending parallel to a longitudinal axis of the vertebral column, the adjustable sleeve having a bottom sleeve portion having a first end with at least two pre-drilled post holes for coupling the bottom sleeve portion to the posts located on the lowermost template and a second end that extends upward toward a top sleeve portion of the adjustable sleeve having a first end with two or more pre-drilled post holes that couple the top sleeve portion to the posts located on the uppermost template, wherein the top sleeve portion extends downward toward the lowermost template and terminates at a second end that forms an opening to a hollow longitudinal channel that extends between the first end and second end of the top sleeve portion, the second end and the hollow sleeve being configured to receive slidably the second end of the bottom sleeve portion; a sliding displacement limiter arrangement located on the anterior surface of the top sleeve portion of the adjustable sleeve and having a tongue portion attached to the anterior surface of the bottom sleeve portion that is positioned in the hollow channel of the top sleeve portion, wherein the tongue extends through and slidably connects to an elongated aperture disposed through the anterior surface of the top sleeve portion, and first and second locking plates each associated with one of said uppermost and lowermost templates, said first and second locking plates each having at least two pre-drilled post holes for engaging with the posts of the lowermost template and the uppermost template and thereby securing the adjustable sleeve to the uppermost template and the lowermost template.\",\n \"10.The implantable mobile sleeve structure of claim 9 further comprising: a second bottom sleeve portion of a second adjustable sleeve placed onto the posts of the uppermost template over the top sleeve portion and under said first locking plate; a second uppermost template having a generally rectangular shape with the lengthwise side of the rectangle extending perpendicular to the axis of a vertebral column, the second uppermost template having an anterior surface, a posterior surface that communicates with a third vertebra, and an outer edge between extending between the anterior surface and the posterior surface, and at least two pre-drilled through-holes penetrating through the second uppermost template between the anterior surface to the posterior surface, wherein the pre-drilled holes are configured to receive two or more fasteners; two posts attached to the anterior surface of the second uppermost template and arranged to extend orthogonally outward therefrom; two fasteners respectively disposed through the two pre-drilled holes of the second uppermost template, each fastener attaching the second uppermost template to the third vertebra, wherein the lengthwise side of the second lowermost template and the lengthwise side of the second uppermost template are parallel when the lowermost template is attached the uppermost template and the second uppermost template is attached to the third vertebra, and a second top sleeve portion of a second adjustable sleeve installed on the posts.\",\n \"11.The implantable mobile sleeve structure of claim 10 wherein the vertebra are part of an anterior cervical vertebral column.\",\n \"12.A method of attaching a mobile sleeve between two adjacent vertebra, the method comprising the steps of: attaching a lowermost template to the anterior vertebral body of a first vertebra of a vertebral column using fasteners disposed through pre-drilled holes in the lowermost template; attaching an uppermost template to the anterior vertebral body of a second vertebra of the vertebral column located above and adjacent to the first vertebra using fasteners disposed through pre-drilled holes in the uppermost template; bending a bottom sleeve portion of an adjustable sleeve to conform to the curvature of the vertebral column; placing a first end of the bottom sleeve portion of the adjustable sleeve using pre-drilled post holes located at the first end of the bottom sleeve portion onto a set of posts that extend outward from the anterior surface of the lowermost template; placing a first end of a top sleeve portion of the adjustable sleeve using pre-drilled post holes located at the first end of the top sleeve portion onto posts that extend out of the anterior surface of the uppermost template, and attaching a locking plate onto the posts of the lowermost template and the uppermost template over the adjustable sleeve and the lowermost template and the uppermost template.\",\n \"13.The method of claim 12 further comprising the further steps of: removing the locking plate located on top of the uppermost template; attaching a second uppermost template of a second mobile sleeve to the anterior vertebral body of a third vertebra located above and adjacent to the second vertebra using fasteners disposed through pre-drilled holes in the second uppermost template; bending a second bottom sleeve portion of a second adjustable sleeve to conform to the curvature of the vertebral column; placing a first end of the second bottom sleeve portion of the second adjustable sleeve using pre-drilled post holes located at the first end the second bottom sleeve portion onto a set of posts that extend outward from the anterior surface of the top plate of the first mobile sleeve; placing a first end of a second top sleeve portion of the second adjustable sleeve using pre-drilled post holes located at the first end the second top sleeve portion onto posts that extend out of the anterior surface of the second uppermost template; attaching a locking plate onto the posts of the uppermost template over the adjustable sleeve and the second adjustable sleeve, and attaching a locking plate onto the posts of the second uppermost template over the adjustable sleeve and the second uppermost template.\",\n \"14.The method of claim 13 wherein the vertebral column is the an anterior cervical column.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>1.Field of the Invention This invention relates generally to spinal implant systems, and more particularly, to a mobile sleeve structure that maintains a determinable spatial relationship between adjacent vertebrae.\",\n \"2.Description of the Related Art There is a need in the field of neurosurgery for spinal implant systems that enable surgeons to maintain a determinable spatial relationship between adjacent vertebrae during vertebral fusion procedures.\",\n \"A typical vertebral fusion procedure involves implanting a bone graft or cage element into the intervertebral region known that normally would be occupied by a disc.\",\n \"Patients that suffer from degenerative disc disorders may be required to undergo a surgical procedure that involves the fusing together ofthe adjacent vertebrae that are located immediately above and below the disc region.\",\n \"During a vertebral fusion procedure, bone grafts are inserted into the disc region to facilitate fusion of the vertebrae by promoting growth of bone in the disc.\",\n \"The result is that the vertebrae will eventually fuse together.\",\n \"Such patients may additionally require a series of such bone grafts that will result in the fusing together of several vertebrae in the cervical vertebral column.\",\n \"A problem with vertebral fusion procedures is that once the bone grafts are implanted they are prone to becoming misplace due to movement of the adjacent vertebrae.\",\n \"In response to this problem several systems have been developed that will immobilize the adjacent vertebrae by attaching a plate between the adjacent vertebrae.\",\n \"The attached plate will maintain the spatial relationship between the vertebrae and promote fusion by preventing slippage of the bone grafts during the fusion process.\",\n \"One known system for maintaining spinal elements in a desired spatial relationship incorporates a series of plates having predetermined heights and shapes.\",\n \"Each such plate has a plurality of apertures disposed at lengthwise distal ends of the plate.\",\n \"Bone screws are threaded through the apertures to attach the plate to a corresponding one of a pair of anterior cervical vertebrae.\",\n \"The plates so attached maintain a spatial relationship between the two adjacent vertebrae that are located immediately above and below a bone graft implant.\",\n \"In the known system, the plates are attached to extend longitudinally with respect to the axis of an anterior cervical vertebral column.\",\n \"It is a deficiency of the above system that plates of various heights must be prefabricated and made available during the procedure because the distance between respective pairs of vertebrae will vary along the length of the anterior cervical column.\",\n \"Thus, upon the conclusion of the procedure there will remain several unused plates that will have to be disposed of or sterilized.\",\n \"Another problem with the known system is that the plates are only used between two vertebrae and cannot be used to maintain a desired spatial relationship between three or more vertebrae.\",\n \"Three or more vertebrae may need to be secured in predetermined relation to one another when the patient is diagnosed to require more than one bone graft implant.\",\n \"Some systems have been developed that use a one-piece rod that can be attached across several vertebrae.\",\n \"A problem with the known rod system is that it does not accommodate the natural lordotic curvature of an individual patient's spinal column as the lordotic curvature varies from patient to patient.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>The foregoing and other objects are achieved by this invention which provides an implantable mobile sleeve for stabilizing the anterior side of a vertebral column.\",\n \"The implantable mobile sleeve has a group ofgenerally rectangular components that generally have an “T” shape.\",\n \"The mobile sleeve has a lowermost template with a generally rectangular shape, in that the lowermost template has two distal length dimensions that are longer than the dimensions of two opposing sides that constitute the width of the plate.\",\n \"When mounted to a vertebra the lengthwise sides of the rectangular shaped lowermost template will extend in a direction substantially perpendicular to the axis of the vertebral column.\",\n \"The lowermost template has a posterior surface that rests against the surface of a vertebra, and an opposing anterior surface that faces outward from the surface of the vertebra.\",\n \"The lowermost template has two or more pre-drilled apertures that penetrate the lowermost template from the posterior surface to the anterior surface.\",\n \"The pre-drilled apertures are configured to receive a fastener to attach the lowermost template to the vertebra.\",\n \"Two posts extend substantially orthogonally outward from the anterior surface.\",\n \"The posts facilitate the fastening of other components of the mobile sleeve to the lowermost template.\",\n \"In some embodiments, the posts are threaded so that a fastener can be used to fix components that are installed on the posts.\",\n \"In one embodiment of the invention, the mobile sleeve is provided with an uppermost template having a generally rectangular shape.\",\n \"The uppermost template is substantially identical to the lowermost template, except that the uppermost template is mounted to a second vertebra located above the first vertebra where the bottom plate is mounted.\",\n \"During attachment, the mobile sleeve is attached between adjacent vertebrae on the vertebral column.\",\n \"The lowermost template will be coupled to a first vertebra and the uppermost template will be coupled to a second vertebra disposed above the first vertebra.\",\n \"In a preferred embodiment, the templates are secured to the anterior region of a cervical vertebral column.\",\n \"However, the mobile sleeve structure described herein may additionally be attached to other regions of the cervical vertebral column, or to other areas of the spine.\",\n \"Both the uppermost template and the lowermost templates are each affixed to vertebrae using two or more fasteners disposed through the two or more pre-drilled through holes located on the top and lowermost templates.\",\n \"The fasteners, in this embodiment, are be inserted into the vertebrae at respective angles in order to prevent the templates from being pulled away from the vertebrae.\",\n \"The fasteners attach the lowermost template to the first vertebra and the uppermost template to the second vertebra.\",\n \"Such fasteners can, in certain embodiments of the invention, be threaded bone screws, pins, or any other suitable fastener for attaching the templates to the vertebrae.\",\n \"While only two bone screws are described herein as being used in the practice of the illustrative embodiment of the invention, persons of skill in the art can, in light of the teaching herein, employ a different number of fasteners depending on conditions encountered during the procedure, such as vertebral size or bone density, spatial positioning of the vertebrae, and the strength of attachment required by the physician.\",\n \"In other embodiments, one or both of the top and lowermost templates are affixed to the vertebrae using adhesives that are applied over the posterior surface of the templates.\",\n \"Once the bottom plate and the uppermost template are attached to the respective first and second vertebrae, it is preferred that the lengthwise sides of the bottom and uppermost templates be parallel.\",\n \"In a further embodiment of the invention, a generally rectangular adjustable sleeve slidably connects the uppermost template and the lowermost template by means of the aforementioned posts and/or apertures through, the templates.\",\n \"It is an advantage of the invention that perfect placement of the adjustable sleeve is achieved because the posts extend generally perpendicular with respect to the midportion of the anterior vertebral body.\",\n \"The adjustable sleeve has a bottom sleeve portion that perpendicularly couples at a first end to the middle region of the lowermost template.\",\n \"A second end of the bottom sleeve portion extends upward toward the uppermost template.\",\n \"With respect to the distance between the anterior and posterior surfaces, the bottom sleeve portion will have a thickness at the first end that differs from that of its second end.\",\n \"In some embodiments of the invention, the thickness at the first end is about 1 mm and will increase to about 2 mm in the middle region and at second end of the bottom sleeve portion.\",\n \"The reason for the varying thickness is so that several adjustable sleeves can be stacked onto the posts of the lowermost template without any large protrusions in the lowermost template area.\",\n \"The thickness of the lowermost template is not limited to 1 mm and 2 mm since sleeves of various thicknesses will be needed depending on the needs of a particular surgical procedure.\",\n \"In a further embodiment, the adjustable sleeve has a top sleeve portion with a first end that couples to post holes in the middle region of the uppermost template.\",\n \"The top sleeve portion also extends downward toward the bottom sleeve portion and terminates at a second end.\",\n \"With regard to thickness (i.e., the distance between the anterior and posterior surfaces), the top sleeve portion will have a different thickness at the first end than at the second end of the top sleeve portion.\",\n \"Illustratively, the thickness at the first end can be about 1 mm and will increase to about 2 mm as for the middle and second end of the top sleeve portion.\",\n \"The reason for the varying thickness is so that several adjustable sleeves can be stacked onto the posts of the uppermost template without any large protrusions in the uppermost template area.\",\n \"The thickness of the uppermost template is not limited to 1 mm and 2 mm since sleeves of various thicknesses will be needed depending on the needs of a particular surgical procedure.\",\n \"The second end of the top sleeve portion also has an opening to a hollow channel that extends upward toward the first end of the top sleeve portion.\",\n \"The second end of the bottom sleeve portion enters the bottom end of the top sleeve portion and is slidably disposed through the hollow channel.\",\n \"The hollow channel affords mobility to the top third of the adjustable sleeve and allows the height of the mobile sleeve to adjust for post-operative subsidence of the vertebrae.\",\n \"The movement of the adjustable sleeve during subsidence enables the resulting load to be shared equally by all of the components of the mobile sleeve.\",\n \"Such distribution of the load increases the integrity of the mobile sleeve by preventing the damage to the templates, adjustable sleeve, or the vertebrae that may occur when the load is not equally distributed.\",\n \"In order to control sliding movement between the top and lowermost templates, a tongue and groove mechanism can, in certain embodiments, be incorporated along the anterior surface of the adjustable sleeve.\",\n \"One or more tongues protrude from the anterior surface of the bottom sleeve portion that is disposed through the hollow channel, extending orthogonally outward from the anterior surface.\",\n \"One or more grooves are located on the anterior surface of the top sleeve portion and are arranged to extend completely through the anterior side of the top sleeve portion, whereby the tongues will extend into and slide freely within the grooves.\",\n \"In the practice of this embodiment, the grooves are of sufficient length to allow a top sleeve portion and a bottom sleeve portion to slide relative to each other and thereby accommodate subsidence of the vertebrae.\",\n \"However, the grooves will not be so long as to permit the bottom sleeve portion to become separated from the top sleeve portion.\",\n \"The lowermost two thirds portion of the adjustable sleeve (i.e., the lower portion of the bottom sleeve portion) is formed of a solid material that can be bent with a plate bender so that the adjustable sleeve will correspond substantially to the curvature of the vertebral column.\",\n \"Although the adjustable sleeve is described herein as having a top third portion that can be adjusted for length and a bottom two thirds that are solid, different ratios are achievable depending upon the region to which the mobile sleeve is to be attached, or the particular needs of a patient.\",\n \"In a further embodiment, a locking plate holds the adjustable sleeve and template together to allow the distribution of stress to all the parts of the mobile sleeve.\",\n \"Controlling the distribution of stress contributes to advantageous distribution of the load by ensuring that no one particular portion of the mobile sleeve is burdened excessively by the stress that is caused by the subsidence of the vertebrae.\",\n \"The locking plate in this embodiment has two or more post holes that are aligned and placed onto the two or more posts of the bottom or uppermost templates.\",\n \"In order to hold the locking plate in place, the post holes of the locking plate are configured to be secured onto the posts as the locking plate moves into place.\",\n \"In other embodiments, the posts of the uppermost template and the lowermost template are threaded so that a fastener, such as a nut, can be used to lock the locking plate in place.\",\n \"In a yet further embodiment of the invention, additional mobile sleeve structures are linked to form additional levels of structure to the spinal support system.\",\n \"The post holes can be used as a way of linking together mobile sleeve structures when a patient needs more than one implant.\",\n \"A second mobile sleeve is linked to a first mobile sleeve by removing the locking plate of either the uppermost template or the lowermost template, depending on whether the site of the next bone graft implant is above or below the uppermost template or the lowermost template of the first mobile sleeve.\",\n \"A second adjustable sleeve is then placed onto the post holes over the first adjustable sleeve and the locking plate is then placed back on to the holes.\",\n \"A second uppermost template or lowermost template (i. e., depending on whether the second sleeve is being place above or below the first mobile sleeve) is then attached to a vertebra so that the second end of the second adjustable sleeve is placed onto the posts of the second uppermost template or the second lowermost template.\",\n \"A locking plate then is placed over either the second uppermost template or the second lowermost template and the second end of the adjustable sleeve.\",\n \"While the second mobile sleeve has been described as attaching so that the mobile sleeves are attached across a series of three adjacent vertebra, the mobile sleeve structure may be attached to three vertebrae that span across more than three vertebrae of the vertebral column.\",\n \"In addition, more levels may be attached to the mobile sleeve structure to create a device that has any multiple number (i.e., two, three, four, etc.)\",\n \"of mobile sleeves in the structure.\",\n \"In accordance with a further aspect of the present invention, there is provided a method of attaching a mobile sleeve between two adjacent cervical vertebrae.\",\n \"The method includes the steps of: attaching a lowermost template to the anterior vertebral body of a first vertebra using fasteners disposed through pre-drilled holes in the lowermost template; attaching an uppermost template to the anterior vertebral body of a second vertebra located above and adjacent to the first vertebra; and installing fasteners through pre-drilled holes in the uppermost template.\",\n \"In a further embodiment of this method aspect of the invention, the first and second vertebrae are separated by one or more vertebrae, and therefore are not adjacent to one another.\",\n \"For instance, the uppermost template and the lowermost template can be attached to respective vertebra in a manner wherein the mobile sleeve spans across any number of vertebrae.\",\n \"In a further embodiment, there is provided the step of bending the bottom sleeve portion of an adjustable sleeve in response to the curvature of the vertebral column.\",\n \"Additionally, there are provided the steps of installing the first end of the bottom sleeve portion of the adjustable sleeve onto posts that extend outward of the anterior surface of the lowermost template, and further installing the first end of a top sleeve portion of the adjustable sleeve is placed onto posts that extend out of the anterior surface of the uppermost template.\",\n \"As previously described, each adjustable sleeve fits onto the posts using pre-drilled post holes located at the first ends of the bottom and top sleeve portions, respectively.\",\n \"In a further embodiment, there is provided the step of coupling the adjustable sleeve to the top and lowermost templates at a generally perpendicular angle.\",\n \"In a further embodiment of this method aspect of the invention, there is provided the step of installing a locking plate onto the posts of the uppermost template and the lowermost template over the adjustable sleeve and the uppermost template and lowermost template.\",\n \"Additionally, there is provided the step of locking the locking plate with a fastener.\",\n \"Illustratively, when a second level (i.e., a second mobile sleeve) is added to a vertebra located above the second vertebra, there is provided a step of removing the locking plate located on the uppermost template.\",\n \"Then, there is performed a step of attaching the second uppermost template of the second mobile sleeve to the anterior vertebral body of a third vertebra located above and adjacent to the second vertebra using fasteners disposed through pre-drilled holes in the uppermost template.\",\n \"In a highly advantageous embodiment of the method aspect of the invention, there is provided the step of bending the bottom sleeve portion of a second adjustable sleeve in response to the curvature of the vertebral column.\",\n \"A first end of the bottom sleeve portion of the second adjustable sleeve is placed onto posts that extend out of the anterior surface of the uppermost template of the first mobile sleeve.\",\n \"The first end of a top sleeve portion of the adjustable sleeve is placed onto posts that extend out of the anterior surface of the second uppermost template attached to the third vertebra.\",\n \"The second adjustable sleeve is configured to fit onto the posts using, pre-drilled post holes located at the first ends of the bottom sleeve portion and top sleeve portion.\",\n \"The adjustable sleeve couples the top and lowermost templates at a generally perpendicular angle.\",\n \"There is additionally provided the step of placing locking plates onto the posts of the uppermost template of the first mobile sleeve and the second uppermost template.\",\n \"In this embodiment, the locking plates are placed over the ends of the second adjustable sleeve and uppermost template of the first mobile sleeve and the second uppermost template.\",\n \"Then, in this embodiment, there is provided the step of affixing the locking plate with a fastener.\",\n \"It is to be understood that once the two mobile sleeve devices have been connected, additional levels can be added to this structure by simply repeating the steps recited for attaching the second mobile sleeve.\",\n \"The mobile sleeves may be coupled in a manner that permits the adjustable sleeve portion to span across more than one vertebrae, as described above.\",\n \"This provides the significant advantage that the interconnection of several levels of mobile sleeves will constitute a support structure that will be custom fitted to the lordotic curvature of the anterior cervical column.\"\n ],\n [\n \"RELATIONSHIP TO OTHER PATENT APPLICATION This application claims the benefit of U.S.\",\n \"Provisional Patent Application No.\",\n \"60/381,579, filed Apr.\",\n \"4, 2001.BACKGROUND OF THE INVENTION 1.Field of the Invention This invention relates generally to spinal implant systems, and more particularly, to a mobile sleeve structure that maintains a determinable spatial relationship between adjacent vertebrae.\",\n \"2.Description of the Related Art There is a need in the field of neurosurgery for spinal implant systems that enable surgeons to maintain a determinable spatial relationship between adjacent vertebrae during vertebral fusion procedures.\",\n \"A typical vertebral fusion procedure involves implanting a bone graft or cage element into the intervertebral region known that normally would be occupied by a disc.\",\n \"Patients that suffer from degenerative disc disorders may be required to undergo a surgical procedure that involves the fusing together ofthe adjacent vertebrae that are located immediately above and below the disc region.\",\n \"During a vertebral fusion procedure, bone grafts are inserted into the disc region to facilitate fusion of the vertebrae by promoting growth of bone in the disc.\",\n \"The result is that the vertebrae will eventually fuse together.\",\n \"Such patients may additionally require a series of such bone grafts that will result in the fusing together of several vertebrae in the cervical vertebral column.\",\n \"A problem with vertebral fusion procedures is that once the bone grafts are implanted they are prone to becoming misplace due to movement of the adjacent vertebrae.\",\n \"In response to this problem several systems have been developed that will immobilize the adjacent vertebrae by attaching a plate between the adjacent vertebrae.\",\n \"The attached plate will maintain the spatial relationship between the vertebrae and promote fusion by preventing slippage of the bone grafts during the fusion process.\",\n \"One known system for maintaining spinal elements in a desired spatial relationship incorporates a series of plates having predetermined heights and shapes.\",\n \"Each such plate has a plurality of apertures disposed at lengthwise distal ends of the plate.\",\n \"Bone screws are threaded through the apertures to attach the plate to a corresponding one of a pair of anterior cervical vertebrae.\",\n \"The plates so attached maintain a spatial relationship between the two adjacent vertebrae that are located immediately above and below a bone graft implant.\",\n \"In the known system, the plates are attached to extend longitudinally with respect to the axis of an anterior cervical vertebral column.\",\n \"It is a deficiency of the above system that plates of various heights must be prefabricated and made available during the procedure because the distance between respective pairs of vertebrae will vary along the length of the anterior cervical column.\",\n \"Thus, upon the conclusion of the procedure there will remain several unused plates that will have to be disposed of or sterilized.\",\n \"Another problem with the known system is that the plates are only used between two vertebrae and cannot be used to maintain a desired spatial relationship between three or more vertebrae.\",\n \"Three or more vertebrae may need to be secured in predetermined relation to one another when the patient is diagnosed to require more than one bone graft implant.\",\n \"Some systems have been developed that use a one-piece rod that can be attached across several vertebrae.\",\n \"A problem with the known rod system is that it does not accommodate the natural lordotic curvature of an individual patient's spinal column as the lordotic curvature varies from patient to patient.\",\n \"SUMMARY OF THE INVENTION The foregoing and other objects are achieved by this invention which provides an implantable mobile sleeve for stabilizing the anterior side of a vertebral column.\",\n \"The implantable mobile sleeve has a group ofgenerally rectangular components that generally have an “T” shape.\",\n \"The mobile sleeve has a lowermost template with a generally rectangular shape, in that the lowermost template has two distal length dimensions that are longer than the dimensions of two opposing sides that constitute the width of the plate.\",\n \"When mounted to a vertebra the lengthwise sides of the rectangular shaped lowermost template will extend in a direction substantially perpendicular to the axis of the vertebral column.\",\n \"The lowermost template has a posterior surface that rests against the surface of a vertebra, and an opposing anterior surface that faces outward from the surface of the vertebra.\",\n \"The lowermost template has two or more pre-drilled apertures that penetrate the lowermost template from the posterior surface to the anterior surface.\",\n \"The pre-drilled apertures are configured to receive a fastener to attach the lowermost template to the vertebra.\",\n \"Two posts extend substantially orthogonally outward from the anterior surface.\",\n \"The posts facilitate the fastening of other components of the mobile sleeve to the lowermost template.\",\n \"In some embodiments, the posts are threaded so that a fastener can be used to fix components that are installed on the posts.\",\n \"In one embodiment of the invention, the mobile sleeve is provided with an uppermost template having a generally rectangular shape.\",\n \"The uppermost template is substantially identical to the lowermost template, except that the uppermost template is mounted to a second vertebra located above the first vertebra where the bottom plate is mounted.\",\n \"During attachment, the mobile sleeve is attached between adjacent vertebrae on the vertebral column.\",\n \"The lowermost template will be coupled to a first vertebra and the uppermost template will be coupled to a second vertebra disposed above the first vertebra.\",\n \"In a preferred embodiment, the templates are secured to the anterior region of a cervical vertebral column.\",\n \"However, the mobile sleeve structure described herein may additionally be attached to other regions of the cervical vertebral column, or to other areas of the spine.\",\n \"Both the uppermost template and the lowermost templates are each affixed to vertebrae using two or more fasteners disposed through the two or more pre-drilled through holes located on the top and lowermost templates.\",\n \"The fasteners, in this embodiment, are be inserted into the vertebrae at respective angles in order to prevent the templates from being pulled away from the vertebrae.\",\n \"The fasteners attach the lowermost template to the first vertebra and the uppermost template to the second vertebra.\",\n \"Such fasteners can, in certain embodiments of the invention, be threaded bone screws, pins, or any other suitable fastener for attaching the templates to the vertebrae.\",\n \"While only two bone screws are described herein as being used in the practice of the illustrative embodiment of the invention, persons of skill in the art can, in light of the teaching herein, employ a different number of fasteners depending on conditions encountered during the procedure, such as vertebral size or bone density, spatial positioning of the vertebrae, and the strength of attachment required by the physician.\",\n \"In other embodiments, one or both of the top and lowermost templates are affixed to the vertebrae using adhesives that are applied over the posterior surface of the templates.\",\n \"Once the bottom plate and the uppermost template are attached to the respective first and second vertebrae, it is preferred that the lengthwise sides of the bottom and uppermost templates be parallel.\",\n \"In a further embodiment of the invention, a generally rectangular adjustable sleeve slidably connects the uppermost template and the lowermost template by means of the aforementioned posts and/or apertures through, the templates.\",\n \"It is an advantage of the invention that perfect placement of the adjustable sleeve is achieved because the posts extend generally perpendicular with respect to the midportion of the anterior vertebral body.\",\n \"The adjustable sleeve has a bottom sleeve portion that perpendicularly couples at a first end to the middle region of the lowermost template.\",\n \"A second end of the bottom sleeve portion extends upward toward the uppermost template.\",\n \"With respect to the distance between the anterior and posterior surfaces, the bottom sleeve portion will have a thickness at the first end that differs from that of its second end.\",\n \"In some embodiments of the invention, the thickness at the first end is about 1 mm and will increase to about 2 mm in the middle region and at second end of the bottom sleeve portion.\",\n \"The reason for the varying thickness is so that several adjustable sleeves can be stacked onto the posts of the lowermost template without any large protrusions in the lowermost template area.\",\n \"The thickness of the lowermost template is not limited to 1 mm and 2 mm since sleeves of various thicknesses will be needed depending on the needs of a particular surgical procedure.\",\n \"In a further embodiment, the adjustable sleeve has a top sleeve portion with a first end that couples to post holes in the middle region of the uppermost template.\",\n \"The top sleeve portion also extends downward toward the bottom sleeve portion and terminates at a second end.\",\n \"With regard to thickness (i.e., the distance between the anterior and posterior surfaces), the top sleeve portion will have a different thickness at the first end than at the second end of the top sleeve portion.\",\n \"Illustratively, the thickness at the first end can be about 1 mm and will increase to about 2 mm as for the middle and second end of the top sleeve portion.\",\n \"The reason for the varying thickness is so that several adjustable sleeves can be stacked onto the posts of the uppermost template without any large protrusions in the uppermost template area.\",\n \"The thickness of the uppermost template is not limited to 1 mm and 2 mm since sleeves of various thicknesses will be needed depending on the needs of a particular surgical procedure.\",\n \"The second end of the top sleeve portion also has an opening to a hollow channel that extends upward toward the first end of the top sleeve portion.\",\n \"The second end of the bottom sleeve portion enters the bottom end of the top sleeve portion and is slidably disposed through the hollow channel.\",\n \"The hollow channel affords mobility to the top third of the adjustable sleeve and allows the height of the mobile sleeve to adjust for post-operative subsidence of the vertebrae.\",\n \"The movement of the adjustable sleeve during subsidence enables the resulting load to be shared equally by all of the components of the mobile sleeve.\",\n \"Such distribution of the load increases the integrity of the mobile sleeve by preventing the damage to the templates, adjustable sleeve, or the vertebrae that may occur when the load is not equally distributed.\",\n \"In order to control sliding movement between the top and lowermost templates, a tongue and groove mechanism can, in certain embodiments, be incorporated along the anterior surface of the adjustable sleeve.\",\n \"One or more tongues protrude from the anterior surface of the bottom sleeve portion that is disposed through the hollow channel, extending orthogonally outward from the anterior surface.\",\n \"One or more grooves are located on the anterior surface of the top sleeve portion and are arranged to extend completely through the anterior side of the top sleeve portion, whereby the tongues will extend into and slide freely within the grooves.\",\n \"In the practice of this embodiment, the grooves are of sufficient length to allow a top sleeve portion and a bottom sleeve portion to slide relative to each other and thereby accommodate subsidence of the vertebrae.\",\n \"However, the grooves will not be so long as to permit the bottom sleeve portion to become separated from the top sleeve portion.\",\n \"The lowermost two thirds portion of the adjustable sleeve (i.e., the lower portion of the bottom sleeve portion) is formed of a solid material that can be bent with a plate bender so that the adjustable sleeve will correspond substantially to the curvature of the vertebral column.\",\n \"Although the adjustable sleeve is described herein as having a top third portion that can be adjusted for length and a bottom two thirds that are solid, different ratios are achievable depending upon the region to which the mobile sleeve is to be attached, or the particular needs of a patient.\",\n \"In a further embodiment, a locking plate holds the adjustable sleeve and template together to allow the distribution of stress to all the parts of the mobile sleeve.\",\n \"Controlling the distribution of stress contributes to advantageous distribution of the load by ensuring that no one particular portion of the mobile sleeve is burdened excessively by the stress that is caused by the subsidence of the vertebrae.\",\n \"The locking plate in this embodiment has two or more post holes that are aligned and placed onto the two or more posts of the bottom or uppermost templates.\",\n \"In order to hold the locking plate in place, the post holes of the locking plate are configured to be secured onto the posts as the locking plate moves into place.\",\n \"In other embodiments, the posts of the uppermost template and the lowermost template are threaded so that a fastener, such as a nut, can be used to lock the locking plate in place.\",\n \"In a yet further embodiment of the invention, additional mobile sleeve structures are linked to form additional levels of structure to the spinal support system.\",\n \"The post holes can be used as a way of linking together mobile sleeve structures when a patient needs more than one implant.\",\n \"A second mobile sleeve is linked to a first mobile sleeve by removing the locking plate of either the uppermost template or the lowermost template, depending on whether the site of the next bone graft implant is above or below the uppermost template or the lowermost template of the first mobile sleeve.\",\n \"A second adjustable sleeve is then placed onto the post holes over the first adjustable sleeve and the locking plate is then placed back on to the holes.\",\n \"A second uppermost template or lowermost template (i. e., depending on whether the second sleeve is being place above or below the first mobile sleeve) is then attached to a vertebra so that the second end of the second adjustable sleeve is placed onto the posts of the second uppermost template or the second lowermost template.\",\n \"A locking plate then is placed over either the second uppermost template or the second lowermost template and the second end of the adjustable sleeve.\",\n \"While the second mobile sleeve has been described as attaching so that the mobile sleeves are attached across a series of three adjacent vertebra, the mobile sleeve structure may be attached to three vertebrae that span across more than three vertebrae of the vertebral column.\",\n \"In addition, more levels may be attached to the mobile sleeve structure to create a device that has any multiple number (i.e., two, three, four, etc.)\",\n \"of mobile sleeves in the structure.\",\n \"In accordance with a further aspect of the present invention, there is provided a method of attaching a mobile sleeve between two adjacent cervical vertebrae.\",\n \"The method includes the steps of: attaching a lowermost template to the anterior vertebral body of a first vertebra using fasteners disposed through pre-drilled holes in the lowermost template; attaching an uppermost template to the anterior vertebral body of a second vertebra located above and adjacent to the first vertebra; and installing fasteners through pre-drilled holes in the uppermost template.\",\n \"In a further embodiment of this method aspect of the invention, the first and second vertebrae are separated by one or more vertebrae, and therefore are not adjacent to one another.\",\n \"For instance, the uppermost template and the lowermost template can be attached to respective vertebra in a manner wherein the mobile sleeve spans across any number of vertebrae.\",\n \"In a further embodiment, there is provided the step of bending the bottom sleeve portion of an adjustable sleeve in response to the curvature of the vertebral column.\",\n \"Additionally, there are provided the steps of installing the first end of the bottom sleeve portion of the adjustable sleeve onto posts that extend outward of the anterior surface of the lowermost template, and further installing the first end of a top sleeve portion of the adjustable sleeve is placed onto posts that extend out of the anterior surface of the uppermost template.\",\n \"As previously described, each adjustable sleeve fits onto the posts using pre-drilled post holes located at the first ends of the bottom and top sleeve portions, respectively.\",\n \"In a further embodiment, there is provided the step of coupling the adjustable sleeve to the top and lowermost templates at a generally perpendicular angle.\",\n \"In a further embodiment of this method aspect of the invention, there is provided the step of installing a locking plate onto the posts of the uppermost template and the lowermost template over the adjustable sleeve and the uppermost template and lowermost template.\",\n \"Additionally, there is provided the step of locking the locking plate with a fastener.\",\n \"Illustratively, when a second level (i.e., a second mobile sleeve) is added to a vertebra located above the second vertebra, there is provided a step of removing the locking plate located on the uppermost template.\",\n \"Then, there is performed a step of attaching the second uppermost template of the second mobile sleeve to the anterior vertebral body of a third vertebra located above and adjacent to the second vertebra using fasteners disposed through pre-drilled holes in the uppermost template.\",\n \"In a highly advantageous embodiment of the method aspect of the invention, there is provided the step of bending the bottom sleeve portion of a second adjustable sleeve in response to the curvature of the vertebral column.\",\n \"A first end of the bottom sleeve portion of the second adjustable sleeve is placed onto posts that extend out of the anterior surface of the uppermost template of the first mobile sleeve.\",\n \"The first end of a top sleeve portion of the adjustable sleeve is placed onto posts that extend out of the anterior surface of the second uppermost template attached to the third vertebra.\",\n \"The second adjustable sleeve is configured to fit onto the posts using, pre-drilled post holes located at the first ends of the bottom sleeve portion and top sleeve portion.\",\n \"The adjustable sleeve couples the top and lowermost templates at a generally perpendicular angle.\",\n \"There is additionally provided the step of placing locking plates onto the posts of the uppermost template of the first mobile sleeve and the second uppermost template.\",\n \"In this embodiment, the locking plates are placed over the ends of the second adjustable sleeve and uppermost template of the first mobile sleeve and the second uppermost template.\",\n \"Then, in this embodiment, there is provided the step of affixing the locking plate with a fastener.\",\n \"It is to be understood that once the two mobile sleeve devices have been connected, additional levels can be added to this structure by simply repeating the steps recited for attaching the second mobile sleeve.\",\n \"The mobile sleeves may be coupled in a manner that permits the adjustable sleeve portion to span across more than one vertebrae, as described above.\",\n \"This provides the significant advantage that the interconnection of several levels of mobile sleeves will constitute a support structure that will be custom fitted to the lordotic curvature of the anterior cervical column.\",\n \"BRIEF DESCRIPTION OF THE DRAWING Comprehension of the invention is facilitated by reading the following detailed description, in conjunction with the annexed drawings, in which: FIG.\",\n \"1a is an isometric representation of a single level embodiment of mobile sleeve structure constructed in accordance with the invention; FIG.\",\n \"1b is a cross-sectional representation taken along line a-a in FIG.\",\n \"1a; FIG.\",\n \"2 is a plan representation of a portion of the cervical column with the mobile sleeve structure mounted between two cervical vertebrae; FIG.\",\n \"3 is a plan representation of the multi-level embodiment of two mobile sleeve structures attached across three cervical vertebrae, and FIG.\",\n \"4 is a side view of the multi-level embodiment of two mobile sleeve structures attached across three cervical vertebrae.\",\n \"DETAILED DESCRIPTION FIG.\",\n \"1a is an isometric representation of the mobile sleeve 10 that is provided with a series of components that when assembled together cause mobile sleeve 10 to have a shape generally in the form of an uppercase letter “I.” A lowermost template 12 and an uppermost template 14 each have an anterior side 16, a posterior side 17 (not shown in this figure), and an outer edge 20 that extends around the perimeter between posterior side 17 and anterior side 16.A respective pair of posts 18 extend orthogonally outward from each of the anterior sides of the lowermost and uppermost templates.\",\n \"Two posts 18 are located on each anterior side 16 of lowermost template 12 and uppermost template 14.Posts 18 are used to connect lowermost template 12 and uppermost template together and, as will be described hereinafter, to facilitate incorporation of additional levels to the overall structure whereby more than two vertebra are to be fused.\",\n \"Lowermost template 12 and uppermost template 14 each have two or more pre-drilled through-holes 13 that extend from the posterior side to anterior side of each template.\",\n \"The through-holes, in this specific illustrative embodiment of the invention, are threaded to receive a threaded fastener 19, which may be a conventional bone screw.\",\n \"Lowermost template 12 and uppermost template 14 are each also provided with two or more post holes 15 that are configured to receive posts when two or more mobile sleeve structures are linked together as will be described in greater detail in FIG.\",\n \"3.Each of lowermost template 12 and uppermost template 14 has a generally rectangular shape with a width that is longer than the height.\",\n \"When installed, the lowermost and uppermost templates are arranged to be substantially parallel.\",\n \"In the specific illustrative embodiment of the invention, it is preferred that lowermost template 12 and uppermost template 14 are curved as shown so that the posterior sides of both templates are concave.\",\n \"The curved posterior side of lowermost template 12 and uppermost template 14 ensures that both templates are substantially flush with the curved surface of the anterior cervical vertebrae (not shown) when affixed thereto as herein described.\",\n \"Conversely, the anterior sides of the lowermost and uppermost templates are convex, and bulky protrusions at the implant site are thereby minimized.\",\n \"Lowermost template 12 and uppermost template 14 are coupled to one another by a generally rectangular shaped adjustable sleeve 22 that extends substantially vertically in the figure and is coupled substantially orthogonally to the lowermost and uppermost templates at its distal ends.\",\n \"As shown, adjustable sleeve 22 is formed of a top sleeve portion 24 and a bottom sleeve portion 26 arranged longitudinally distal, and each such portion has respective pre-drilled post holes.\",\n \"Bottom sleeve portion 26 is connected to lowermost template 12 by registering the posts located on the anterior surface of lowermost template 12 through the post holes.\",\n \"Similarly, top sleeve portion 24 is connected to uppermost template 14 by registering the posts located on the anterior surface of the uppermost template 14 through the corresponding post holes.\",\n \"Top sleeve portion 24 and bottom sleeve portion 26 each have a generally rectangular shape and may be slidably connected to their respectively associated templates.\",\n \"As shown in FIG.\",\n \"1b, which is cross-sectional representation taken at section line a-a of FIG.\",\n \"la shows that top sleeve portion 24 has an internal region 28 that has a rectangular shape and extends longitudinally.\",\n \"Top sleeve portion 24 slides into the end of, and along, hollow channel 28 which cross-sectionally surrounds same.\",\n \"In order to control the extent of the sliding extension between uppermost template 14 and lowermost template 12 there is provided in this specific illustrative embodiment of the invention, a pair of tongues 27 and corresponding elongated apertures 29 that accommodate the tongues as shown.\",\n \"Tongues 27 are attached to the anterior surface ofthe bottom sleeve portion 26 portion that is disposed through hollow channel 28.Tongues 27 will extend substantially orthogonal outward from the anterior surface.\",\n \"The two elongated apertures 29 are located on the anterior surface of top sleeve portion 24.Elongated apertures 29 extend completely through the anterior side of top sleeve portion 24 so that tongues 27 will extend therethrough and slide freely with respect thereto.\",\n \"The elongated apertures are of sufficient length along their respective major axes to allow top sleeve portion 24 and bottom sleeve portion 26 to be longitudinally movable relative to each other to accommodate subsidence of the vertebrae.\",\n \"In this specific illustrative embodiment of the invention, a locking plate 30 is installed on the posts of the uppermost template.\",\n \"Locking plate 30 serves to retain adjustable sleeve 22 and uppermost template 14 in fixed relation to each other.\",\n \"The locking plate has two pre-drilled post holes 15 that enable the installation of locking plate 30 on the two posts of uppermost template 14.In some embodiments of the invention, a locking plate 30 similar to that described hereinabove is affixed to lowermost template 12.In order to hold the locking plate on the post holes, a known grasping arrangement, such as a resilient locking element (not shown), or a threaded fastened (not shown) can 25 be employed.\",\n \"FIG.\",\n \"2 is a plan representation of a portion of the cervical column with mobile sleeve 10 mountedbetweentwo cervical vertebrae 34 and 36 (represented schematically).\",\n \"Elements of structure that correspond to those discussed in connection with FIGS.\",\n \"1a and 1b are similarly designated.\",\n \"The cervical portion of a human spinal column (not shown) has a bone graft 32 placed between first vertebra 36, located below bone graft 32, and a second vertebra 36, located above bone graft 32.Mobile sleeve 10 is coupled to first vertebra 36 and extends upward longitudinally with respect to cervical column to the region where mobile sleeve 10 attaches to second vertebra 36.Mobile sleeve 10 holds first vertebra 34 and second vertebra 36 in a desired spatial relationship relative to each other.\",\n \"Maintaining the spatial relationship of first vertebra 34 and second vertebra 36 will increase the likelihood of success of spinal fusion procedures by preventing bone graft 32 from shifting and becoming dislodged.\",\n \"Lowermost template 12 is shown to be coupled to first vertebra 36 by a pair of bone screws 38 that are disposed though pre-drilled through-holes in lowermost template 12 discussed above in connection with FIG.\",\n \"la.\",\n \"Uppermost template 14 is shown to be attached to second vertebra 36 with a similar pair of bone screws 38 that are disposed though corresponding pre-drilled through-holes in uppermost template 14.FIGS.\",\n \"3 and 4 are a plan representation and a side partially cross-sectional representation, respectively, of a multi-level embodiment of two mobile sleeve structures attached across three cervical vertebrae.\",\n \"Elements of structure that correspond to those in FIGS.\",\n \"1a, 1b, and 2 are similarly designated.\",\n \"The cervical portion of a human spinal column having first vertebra 34, second vertebra 36, and third vertebra 40 is shown to have two bone grafts 32, 32′.\",\n \"Bone graft 32 is dispose between first vertebra 34 and second vertebra 36, and second bone graft 32′ is disposed between second vertebra 36 and third vertebra 40.Mobile sleeve 10 is attached between first vertebra 34 and second vertebra 36 as described in FIG.\",\n \"2.In this specific illustrative embodiment of the invention, a second level in the form of a second mobile sleeve 10′ is attached between second vertebra 36 and third vertebra 40.A lower sleeve portion 26′ of second mobile sleeve 10′ is aligned and connected to posts 18 of uppermost template 14 of first mobile sleeve 10.Referring to FIG.\",\n \"4 a locking plate 30 is attached to post 18 over bottom sleeve 26′ and uppermost template 14 just as described in FIG.\",\n \"2.Uppermost template 14′ of second mobile sleeve 10 is attached with bone screws 38 to third vertebra 40 located above second bone graft 32′.\",\n \"A top sleeve portion 24′ 30 of second mobile sleeve 10′ is placed on to posts 18 of uppermost template.\",\n \"Uppermost template 14′ of second mobile sleeve 10′ is can be used, in other embodiments of the invention, for placement of yet a third mobile sleeve (not shown), if needed in the future.\",\n \"Although the invention has been described in terms of specific embodiments and applications, persons skilled in the art can, in light of this teaching, generate additional embodiments without exceeding the scope or departing from the spirit of the invention described herein.\",\n \"Accordingly, it is to be understood that the drawing and description in this disclosure are proffered to facilitate comprehension of the invention, and should not be construed to limit the scope thereof.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473759"}}},{"rowIdx":749,"cells":{"text":{"kind":"list like","value":[["Automatic optimization of a splice loss estimator for optical fiber splicers","A technique is provided for automatic optimization of a splice loss estimator of a fiber splicer (1), where the splice loss estimator is adapted, in a splice loss estimation procedure, to estimate the splice losses (Lti) of splices (i) of optical fibers as produced by the fiber splicer from images taken of the optical fibers at the splicing thereof, and the splice loss estimation procedure includes the use of splice loss estimation parameters (Pj).","The estimator estimates splice losses based on information (Cij) obtained from the images and the estimation parameters.","Further, the splice losses are measured by means of a measurement instrument (3).","The estimated (Lti) and measured (LMi) splice losses, and the information obtained from the images are uploaded (71) into an off-line computer (5) and the key estimation parameters are automatically optimized by the selection of any solution within the Bellcore accuracy criteria (75), whereafter the optimized estimation parameters are downloaded (81) to the splicer."],["1.An apparatus for automatic optimization of a splice loss estimator of an optical fiber splicer apparatus, said splice loss estimator being adapted, in a splice loss estimation procedure, to estimate the splice losses (LTi) of splices (i) of end portions of optical fibers as produced by said fiber splicer apparatus from at least one respective image taken of the respective end portions of the optical fibers prior to, during, or subsequent to the splicing thereof, said splice loss estimation procedure including the use of a set of splice loss estimation parameters (Pj), comprising a first input line for receiving, from said splice loss estimator, data of a plurality of splices of end portions of optical fibers as produced by said fiber splicer apparatus, wherein said data for each splice (i) include information (Cij) derived from the at least one image taken of the end portions of the optical fibers constituting that splice, and the estimated splice loss (LTi) of that splice; a second input line for receiving a splice loss value (LMi) for each of said plurality of splices of end portions of optical fibers as measured by a measurement equipment; means for determining a new set of splice loss estimation parameters from said data and said estimated and measured splice loss values such that said splice loss estimation procedure, using the new set of splice loss estimation parameters, estimates the splice loss of each of said plurality of splices of end portions of optical fibers with a given accuracy; and an output line for outputting said determined new set of splice loss estimation parameters to said splice loss estimator to replace the set of splice loss estimation parameters used in the splice loss estimation procedure of the splice loss estimator for the estimation of the splice losses of any further splices of end portions of optical fibers to be produced by said fiber splicer apparatus.","2.The apparatus as claimed in claim 1 wherein said data for each splice (i) include information (Cij) derived from hot images taken of the end portions of the optical fibers constituting that splice.","3.The apparatus as claimed in claim 2 wherein said data for each splice include information regarding mismatch of mode field diameter (MFD), including micro bending loss (LD), or fiber core/cladding deformation including cladding-to-core eccentricities, non-circularity of fibers, cleave angle of fiber ends, and/or cladding/core offsets.","4.The apparatus as claimed in claim 2 wherein said data for each splice include information regarding mismatch of hot core index (LI), including hot core index offset (V), fiber dopant concentrations, and/or fiber core diameters.","5.The apparatus as claimed in claim 1 wherein said data for each splice (i) include information (Cij) derived from cold images taken of the end portions of the optical fibers constituting that splice.","6.The apparatus as claimed in claim 5 wherein said data for each splice include information regarding macro bending, including view angle offset, fiber alignment, and/or presence of microdust on fiber surfaces.","7.The apparatus as claimed in claim 1 wherein said apparatus is a computer, preferably an off-line computer and/or a PC-platform based splicer, provided with suitable software and information as regards the splice loss estimation procedure used in the splice loss estimator.","8.The apparatus as claimed in claim 1 wherein said set of splice loss estimation parameters include any of an M factor, an F factor, a loss factor, a macro factor, a mode field diameter left (MFDL) factor, a mode field diameter right (MFDR) factor, and a splice loss shift (LS) factor, where the M and F factors are employed in the estimation of a core deformation induced splice loss (LM), the loss factor is employed in the estimation of a hot core index mismatch induced splice loss (LI) the macro factor, the mode field diameter left (MFDL) and right (MFDR) factors are employed in the estimation of a macro bending induced splice loss (LA) and the splice loss shift (LS) factor is employed in the estimation of a splice loss correction term, which corrects for any systematic errors in the estimation procedure.","9.The apparatus as claimed in claim 1 wherein said apparatus is adapted to receive, on said first input line, the set of estimation parameters as used by said splicer apparatus.","10.The apparatus as claimed in claim 1 wherein said apparatus is adapted to receive, on said first input line, a set of image processing parameters, wherein said set of image processing parameters are used in said splice loss estimation procedure by said splice loss estimator of the splicer apparatus; said means for determining is adapted to determine a new set of image processing parameters from said data and said estimated and measured splice loss values such that said splice loss estimation procedure, using the new set of image processing parameters, estimates the splice loss of each of said plurality of splices of end portions of optical fibers with a given accuracy; and said apparatus for automatic optimization is adapted to output, via said output line, said determined new set of image processing parameters to said splice estimator to replace the set of image processing parameters used in the splice loss estimation procedure of the splice estimator for the estimation of the splice losses of any further splices of end portions of optical fibers to be produced by said fiber splicer apparatus.","11.The apparatus as claimed in claim 1 wherein said plurality of splices are at least 50, more preferably at least 100, and most preferably between 100 and 500.12.The apparatus as claimed in claim 1 wherein said means for determining is adapted to determine said new set of splice loss estimation parameters such that said new set of splice loss estimation parameters fulfils the Bellcore splice loss estimator criteria, preferably the 90% Bellcore criteria, and most preferably the 100% Bellcore criteria.","13.The apparatus as claimed in claim 1 wherein said means for determining is adapted to determine said new set of splice loss estimation parameters by using linear regression criteria to increase the probability that the estimated splice loss values using said new set of estimation parameters represent the measured splice losses.","14.","(canceled) 15.A method for automatic optimization of a splice loss estimator of an optical fiber splicer apparatus, where said splice loss estimator is adapted, in a splice loss estimation procedure, to estimate the splice losses (LTi) of splices (i) of end portions of optical fibers as produced by said fiber splicer apparatus from at least one respective image taken of the respective end portions of the optical fibers prior to, during, or subsequent to the splicing thereof, said splice loss estimation procedure including the use of a set of splice loss estimation parameters (Pj), comprising the steps of: receiving, from said splice loss estimator, data of a plurality of splices of end portions of optical fibers as produced by said fiber splicer apparatus, wherein said data for each splice (i) include information (Cij) derived from the at least one image taken of the end portions of the optical fibers constituting that splice, and the estimated splice loss (LTi) of that splice; receiving a splice loss value (LMi) for each of said plurality of splices of end portions of optical fibers as measured by a measurement equipment; determining a new set of splice loss estimation parameters from said data and said estimated and measured splice loss values such that said splice loss estimation procedure, using the new set of splice loss estimation parameters, estimates the splice loss of each of said plurality of splices of end portions of optical fibers with a given accuracy; and outputting said determined new set of splice loss estimation parameters to said splice loss estimator to replace the set of splice loss estimation parameters used in the splice loss estimation procedure of the splice loss estimator for the estimation of the splice losses of any further splices of end portions of optical fibers to be produced by said fiber splicer apparatus.","16.The method as claimed in claim 15 wherein said data for each splice (i) include information (Cij) derived from hot images taken of the end portions of the optical fibers constituting that splice.","17.The method as claimed in claim 15 wherein said data for each splice (i) include information (Cij) derived from cold images taken of the end portions of the optical fibers constituting that splice.","18.The method as claimed in claim 15 wherein said set of splice loss estimation parameters include any of an M factor, an F factor, a loss factor, a macro factor, a mode field diameter left (MFDL) factor, a mode field diameter right (MFDR) factor, and a splice loss shift (LS) factor, where the M and F factors are employed in the estimation of a core deformation induced splice loss (LM), the loss factor is employed in the estimation of a hot core index mismatch induced splice loss (LI) the macro factor, the mode field diameter left (MFDL) and right (MFDR) factors are employed in the estimation of a macro bending induced splice loss (LA) and the splice loss shift (LS) factor is employed in the estimation of a splice loss correction term, which corrects for any systematic errors in the estimation procedure.","19.The method as claimed in claim 15 further comprising the steps of: receiving, from said splice estimator, a set of image processing parameters, wherein said set of image processing parameters are used in said splice loss estimation procedure by said splice loss estimator of the splicer apparatus; determining a new set of image processing parameters from said data and said estimated and measured splice loss values such that said splice loss estimation procedure, using the new set of image processing parameters, estimates the splice loss of each of said plurality of splices of end portions of optical fibers with a given accuracy; and outputting said determined new set of image processing parameters to said splice estimator to replace the set of image processing parameters used in the splice loss estimation procedure of the splice estimator for the estimation of the splice losses of any further splices of end portions of optical fibers to be produced by said fiber splicer apparatus.","20.The method as claimed in claim 15 wherein said new set of splice loss estimation parameters is determined such that said new set of splice loss estimation parameters fulfils the Bellcore splice loss estimator criteria, preferably the 90% Bellcore criteria, and most preferably the 100% Bellcore criteria.","21.The method as claimed in claim 15 wherein said new set of splice loss estimation parameters is determined by using linear regression criteria to increase the probability that the estimated splice loss values using said new set of estimation parameters represent the measured splice losses.","22.","(canceled) 23.A method for automatic optimization of a splice loss estimator of an optical fiber splicer apparatus, where said splice loss estimator is adapted, in a splice loss estimation procedure, to estimate the splice losses (LTi) Of splices of end portions of optical fibers as produced by said fiber splicer apparatus from at least one respective image taken of the respective end portions of the optical fibers prior to, during, or subsequent to the splicing thereof, said splice loss estimation procedure including the use of a set of splice loss estimation parameters (Pj), comprising the steps of: producing a plurality of splices of end portions of optical fibers by means of said fiber splicer apparatus; deducing information from the at least one image taken for each splice produced; estimating the splice loss based on the information deduced from the at least one image for each splice produced; measuring the splice loss of each splice produced by means of a measurement instrument,; uploading the deduced information, the estimated splice losses and the measure splice losses to a processing device; determining a new set of splice loss estimation parameters from the deduced information and the estimated and measured splice loss values such that said splice loss estimation procedure, using the new set of splice loss estimation parameters, estimates the splice loss of each of said plurality of splices of end portions of optical fibers with a given accuracy; and downloading said determined new set of splice loss estimation parameters to said splice loss estimator of the fiber splicer apparatus to replace the set of splice loss estimation parameters used in the splice loss estimation procedure of the splice loss estimator for the estimation of the splice losses of any further splices of end portions of optical fibers to be produced by said fiber splicer apparatus."],["<SOH> DESCRIPTION OF RELATED ART AND BACKGROUND OF THE INVENTION <EOH>The technology for forming low-loss optical fibers has advanced to a point where there is widespread commercial manufacturing of optical fibers.","Most processing includes drawing an optical fiber from a previously manufactured glass boule, sometimes referred to as a fiber preform.","Extremely long lengths of fiber can be obtained by splicing a plurality of lengths, which are obtained using current fusion splicer techniques.","Additionally, it has become increasingly more common to splice optical fibers, which have broken, either accidentally, or during appropriate proof testing.","For these and other applications, splicing in which the coating material is removed from end portions of two fibers, which are then fused together end to end, provides a suitable means for joining the ends of two glass fibers with an acceptably low splice loss.","Fusion splice losses depend mainly on the mismatch of mode-field diameter (MFD) and misalignment of the cladding/core of two fibers due to either the fiber refractive index design or the inappropriate fusion processes being used.","With the help of image techniques, information on the MFD mismatch can be obtained and analyzed by monitoring the deformation/misalignment of the cladding/core of the fibers during the fusion process, see e.g.","EP 1 014 070.Such a monitoring system includes typically a charged-coupled device (CCD) camera equipped with an image processor.","By use of suitable theoretical models and the information obtained from the images, an estimation method for evaluation of splice losses can be established.","The estimation method as a passive technique for evaluation of splice losses is widely used in most automated fusion splicers of today.","Different models for evaluation of splice loss have been explored and developed during the past two decades.","Well-known methods for splice loss estimation include the butt-joint approximation and the mode coupling theory.","More advanced methods, e.g.","hot-image techniques for real-time analysis of cladding and core deformation have also been developed.","In order to achieve the best performance and maximize the flexibility of the estimation method, the models used for splice loss estimation include usually a number of free parameters, called estimation parameters.","The optimization of these estimation parameters in order to achieve the best performance of the estimation method has nowadays become one of essential features in the development of splicing techniques.","In practice, the estimation parameters are manually optimized according to different types of on-test fibers and the fusion processes being used.","Due to primarily technical reasons and rather complicated nature of the involved splicing processes, the optimization of the estimation parameters is a quite difficult and time consuming job that may only be performed by very experienced engineers.","The time typically needed for manually optimizing the estimation parameters on a given fiber combination is as long as a few days which is hardly to be accepted by splicer users, especially, when frequently changes of fusion processes and fiber combinations are involved, e.g.","in the manufacture of erbium doped fiber amplifiers (EDFA).","This is hardly acceptable for the splicer user, and such optimization has thus typically to be performed by the fiber splicer manufacturer."],["<SOH> SUMMARY OF THE INVENTION <EOH>The present inventors have realized that the splicer user does not need to re-optimize the image processing parameters, but only the estimation parameters.","For a fiber splicer user, only a limited number of the image processing parameters can be optimized, and most of these parameters are already designed to be linear to the splice loss by an embedded algorithm in the splicer.","In the present invention a new technique for fast automatic optimization of the performance of the estimator is proposed.","The technique leaves the image processing of the cold and hot images taken during the fusion processes unchanged.","The built-in estimator in the splicer analyzes the causes of splice losses based on the information obtained from the images, and based on the fusion process employed and the kind of spliced fibers.","Further, splice losses are measured by means of a measurement instrument, e.g.","a power meter.","The results of the analysis and the measured splice losses are uploaded into a spreadsheet-based worksheet in an off-line computer and/or a PC-platform based splicer to form a splice database.","Then, the manual procedures for optimization of estimation parameters are simulated.","The key estimation parameters are automatically optimized by the selection of any solution within the Bellcore accuracy criteria, or the best solution within the Bellcore accuracy criteria by assistance of regression analysis criteria, whereafter the optimized estimation parameters are downloaded to splicer.","The time needed for optimizing a splice database with hundred splices is significantly reduced down to a few seconds only.","Thus, an enhanced performance of the estimator is automatically achieved.","The tedious and complicated work for manually optimizing the estimator is automated.","It is a main object of the present invention to provide an apparatus for automatic optimization of a splice loss estimator of an optical fiber splicer, wherein the splice loss estimator is adapted, in a splice loss estimation procedure, to estimate the splice losses of splices of end portions of optical fibers as produced by said fiber splicer apparatus from at least one respective image taken of the respective end portions of the optical fibers prior to, during, or subsequent to the splicing thereof, and wherein the splice loss estimation procedure includes the use of a set of splice loss estimation parameters, which apparatus for automatic optimization is fast, and which is particularly suited to be used by a fiber splicer user.","It is in this respect a particular object of the invention to provide such apparatus, which is simple, easy to use, accurate, precise and of low cost.","These objects, among others, are according to a first aspect of the present invention attained by an apparatus comprising a first input line; a second input line; means for determining (preferably a personal computer provided with suitable software); and an output line.","The first input line is provided for receiving, from the splice loss estimator, data of a plurality of splices of end portions of optical fibers as produced by the fiber splicer apparatus, wherein the data for each splice include information derived from the image taken of the end portions of the optical fibers constituting that splice, and the estimated splice loss of that splice; and the second input line is provided for receiving a splice loss value for each of the plurality of splices of end portions of optical fibers as measured by a measurement equipment.","The means for determining is adapted to determine or calculate a new set of splice loss estimation parameters from the data and the measured splice loss values such that the splice loss estimation procedure, using the new set of splice loss estimation parameters, estimates the splice loss of each of the plurality of splices of end portions of optical fibers with a given accuracy.","Finally, the output line is provided for outputting the determined or calculated new set of splice loss estimation parameters to the splice loss estimator of the fiber splicer apparatus to replace the set of splice loss estimation parameters used in the splice loss estimation procedure of the splice loss estimator for the estimation of the splice losses of any further splices of end portions of optical fibers to be produced by the fiber splicer apparatus.","Further, the above-mentioned objects, among others, are according to a second aspect of the present invention attained by an optical fiber splicer system comprising such an apparatus for automatic optimization.","It is a further object of the present invention to provide a method for automatic optimization of the above-depicted splice loss estimator, which is fast and which is thus suitable to be used by the fiber splicer user.","This object is according to a third aspect of the present invention attained by a method comprising the following steps: (i) receiving, from said splice loss estimator, data of a plurality of splices of end portions of optical fibers as produced by said fiber splicer apparatus, wherein said data for each splice include information derived from the at least one image taken of the end portions of the optical fibers constituting that splice, and the estimated splice loss of that splice; (ii) receiving a splice loss value for each of said plurality of splices of end portions of optical fibers as measured by a measurement equipment; (iii) determining a new set of splice loss estimation parameters from said data and said measured splice loss values such that said splice loss estimation procedure, using the new set of splice loss estimation parameters, estimates the splice loss of each of said plurality of splices of end portions of optical fibers with a given accuracy; and (iv) outputting said determined new set of splice loss estimation parameters to said splice loss estimator to replace the set of splice loss estimation parameters used in the splice loss estimation procedure of the splice loss estimator for the estimation of the splice losses of any further splices of end portions of optical fibers to be produced by said fiber splicer apparatus.","Finally, according to a fourth aspect of the present invention there is provided a computer program product loadable into the internal memory of a computer, which comprises software code portions for performing the above-described method when the computer program product is run on the computer.","Further characteristics of the invention and advantages thereof will be evident from the detailed description of preferred embodiments of the invention given hereinbelow and the accompanying drawings, which are given by way of illustration only, and thus are not limitative of the present invention."],["TECHNICAL FIELD OF THE INVENTION The present invention generally relates to fiber optics.","More specifically, the invention relates to -an apparatus and a method, respectively, for automatic optimization of a splice loss estimator for optical fiber splicers, to an optical fiber splicer system comprising such an apparatus, and to a computer program product for performing such a method.","DESCRIPTION OF RELATED ART AND BACKGROUND OF THE INVENTION The technology for forming low-loss optical fibers has advanced to a point where there is widespread commercial manufacturing of optical fibers.","Most processing includes drawing an optical fiber from a previously manufactured glass boule, sometimes referred to as a fiber preform.","Extremely long lengths of fiber can be obtained by splicing a plurality of lengths, which are obtained using current fusion splicer techniques.","Additionally, it has become increasingly more common to splice optical fibers, which have broken, either accidentally, or during appropriate proof testing.","For these and other applications, splicing in which the coating material is removed from end portions of two fibers, which are then fused together end to end, provides a suitable means for joining the ends of two glass fibers with an acceptably low splice loss.","Fusion splice losses depend mainly on the mismatch of mode-field diameter (MFD) and misalignment of the cladding/core of two fibers due to either the fiber refractive index design or the inappropriate fusion processes being used.","With the help of image techniques, information on the MFD mismatch can be obtained and analyzed by monitoring the deformation/misalignment of the cladding/core of the fibers during the fusion process, see e.g.","EP 1 014 070.Such a monitoring system includes typically a charged-coupled device (CCD) camera equipped with an image processor.","By use of suitable theoretical models and the information obtained from the images, an estimation method for evaluation of splice losses can be established.","The estimation method as a passive technique for evaluation of splice losses is widely used in most automated fusion splicers of today.","Different models for evaluation of splice loss have been explored and developed during the past two decades.","Well-known methods for splice loss estimation include the butt-joint approximation and the mode coupling theory.","More advanced methods, e.g.","hot-image techniques for real-time analysis of cladding and core deformation have also been developed.","In order to achieve the best performance and maximize the flexibility of the estimation method, the models used for splice loss estimation include usually a number of free parameters, called estimation parameters.","The optimization of these estimation parameters in order to achieve the best performance of the estimation method has nowadays become one of essential features in the development of splicing techniques.","In practice, the estimation parameters are manually optimized according to different types of on-test fibers and the fusion processes being used.","Due to primarily technical reasons and rather complicated nature of the involved splicing processes, the optimization of the estimation parameters is a quite difficult and time consuming job that may only be performed by very experienced engineers.","The time typically needed for manually optimizing the estimation parameters on a given fiber combination is as long as a few days which is hardly to be accepted by splicer users, especially, when frequently changes of fusion processes and fiber combinations are involved, e.g.","in the manufacture of erbium doped fiber amplifiers (EDFA).","This is hardly acceptable for the splicer user, and such optimization has thus typically to be performed by the fiber splicer manufacturer.","SUMMARY OF THE INVENTION The present inventors have realized that the splicer user does not need to re-optimize the image processing parameters, but only the estimation parameters.","For a fiber splicer user, only a limited number of the image processing parameters can be optimized, and most of these parameters are already designed to be linear to the splice loss by an embedded algorithm in the splicer.","In the present invention a new technique for fast automatic optimization of the performance of the estimator is proposed.","The technique leaves the image processing of the cold and hot images taken during the fusion processes unchanged.","The built-in estimator in the splicer analyzes the causes of splice losses based on the information obtained from the images, and based on the fusion process employed and the kind of spliced fibers.","Further, splice losses are measured by means of a measurement instrument, e.g.","a power meter.","The results of the analysis and the measured splice losses are uploaded into a spreadsheet-based worksheet in an off-line computer and/or a PC-platform based splicer to form a splice database.","Then, the manual procedures for optimization of estimation parameters are simulated.","The key estimation parameters are automatically optimized by the selection of any solution within the Bellcore accuracy criteria, or the best solution within the Bellcore accuracy criteria by assistance of regression analysis criteria, whereafter the optimized estimation parameters are downloaded to splicer.","The time needed for optimizing a splice database with hundred splices is significantly reduced down to a few seconds only.","Thus, an enhanced performance of the estimator is automatically achieved.","The tedious and complicated work for manually optimizing the estimator is automated.","It is a main object of the present invention to provide an apparatus for automatic optimization of a splice loss estimator of an optical fiber splicer, wherein the splice loss estimator is adapted, in a splice loss estimation procedure, to estimate the splice losses of splices of end portions of optical fibers as produced by said fiber splicer apparatus from at least one respective image taken of the respective end portions of the optical fibers prior to, during, or subsequent to the splicing thereof, and wherein the splice loss estimation procedure includes the use of a set of splice loss estimation parameters, which apparatus for automatic optimization is fast, and which is particularly suited to be used by a fiber splicer user.","It is in this respect a particular object of the invention to provide such apparatus, which is simple, easy to use, accurate, precise and of low cost.","These objects, among others, are according to a first aspect of the present invention attained by an apparatus comprising a first input line; a second input line; means for determining (preferably a personal computer provided with suitable software); and an output line.","The first input line is provided for receiving, from the splice loss estimator, data of a plurality of splices of end portions of optical fibers as produced by the fiber splicer apparatus, wherein the data for each splice include information derived from the image taken of the end portions of the optical fibers constituting that splice, and the estimated splice loss of that splice; and the second input line is provided for receiving a splice loss value for each of the plurality of splices of end portions of optical fibers as measured by a measurement equipment.","The means for determining is adapted to determine or calculate a new set of splice loss estimation parameters from the data and the measured splice loss values such that the splice loss estimation procedure, using the new set of splice loss estimation parameters, estimates the splice loss of each of the plurality of splices of end portions of optical fibers with a given accuracy.","Finally, the output line is provided for outputting the determined or calculated new set of splice loss estimation parameters to the splice loss estimator of the fiber splicer apparatus to replace the set of splice loss estimation parameters used in the splice loss estimation procedure of the splice loss estimator for the estimation of the splice losses of any further splices of end portions of optical fibers to be produced by the fiber splicer apparatus.","Further, the above-mentioned objects, among others, are according to a second aspect of the present invention attained by an optical fiber splicer system comprising such an apparatus for automatic optimization.","It is a further object of the present invention to provide a method for automatic optimization of the above-depicted splice loss estimator, which is fast and which is thus suitable to be used by the fiber splicer user.","This object is according to a third aspect of the present invention attained by a method comprising the following steps: (i) receiving, from said splice loss estimator, data of a plurality of splices of end portions of optical fibers as produced by said fiber splicer apparatus, wherein said data for each splice include information derived from the at least one image taken of the end portions of the optical fibers constituting that splice, and the estimated splice loss of that splice; (ii) receiving a splice loss value for each of said plurality of splices of end portions of optical fibers as measured by a measurement equipment; (iii) determining a new set of splice loss estimation parameters from said data and said measured splice loss values such that said splice loss estimation procedure, using the new set of splice loss estimation parameters, estimates the splice loss of each of said plurality of splices of end portions of optical fibers with a given accuracy; and (iv) outputting said determined new set of splice loss estimation parameters to said splice loss estimator to replace the set of splice loss estimation parameters used in the splice loss estimation procedure of the splice loss estimator for the estimation of the splice losses of any further splices of end portions of optical fibers to be produced by said fiber splicer apparatus.","Finally, according to a fourth aspect of the present invention there is provided a computer program product loadable into the internal memory of a computer, which comprises software code portions for performing the above-described method when the computer program product is run on the computer.","Further characteristics of the invention and advantages thereof will be evident from the detailed description of preferred embodiments of the invention given hereinbelow and the accompanying drawings, which are given by way of illustration only, and thus are not limitative of the present invention.","BRIEF DESCRIPTION OF THE DRAWINGS FIG.","1 illustrates, schematically, in a block diagram, an optical fiber splicer system comprising an optical fiber fusion splicer, measurement instruments, and an apparatus for automatic off-line optimization of a splice loss estimator for the optical fiber fusion splicer according to a general embodiment of the present invention.","FIG.","2 illustrates schematically procedures for deriving splice losses.","FIG.","3 illustrates the result of a splice loss estimation procedure, where the Bellcore criteria are fulfilled, which estimation procedure uses a new set of splice loss estimation parameters resulting from the automatic optimization according to the present invention.","FIGS.","4-6 illustrate the results of splice loss estimation procedures using the original set of estimation parameters (FIG.","4); and new sets of splice loss estimation parameters resulting from the automatic optimization according to the present invention (FIGS.","5-6), respectively.","FIG.","7 is a schematic flow chart illustrating an off-line tuning procedure according to a preferred embodiment of the present invention.","DETAILED DESCRIPTION OF EMBODIMENTS In the following description, for purposes of explanation and not limitation, specific details are set forth, such as particular techniques and applications in order to provide a thorough understanding of the present invention.","However, it will be apparent to one skilled in the art that the present invention may be practiced in other embodiments that depart from these specific details.","Optical Fiber Splicer System Including a PC-Based Platform for Supporting Off-Line Tuning With reference to FIG.","1, which schematically illustrates an optical fiber splicer system, a general embodiment of the present invention will be overviewed.","The system comprises an optical fiber fusion splicer 1 including a splice loss estimator (not explicitly illustrated in FIG.","1) for, in a splice loss estimation procedure, estimating the splice losses of splices of end portions of optical fibers as produced by the fiber splicer 1 from at least one respective image taken of the respective end portions of the optical fibers prior to, during, or subsequent to the splicing thereof, where the splice loss estimation procedure includes the use of a set of splice loss estimation parameters.","The optical fiber fusion splicer may e.g.","be an Ericsson 995 Fusion Splicer.","Further, there is provided measurement equipment 3 for measuring the splice loss for a number of splices produced.","The measurement equipment may be a power meter, an extinction ratio meter, or similar.","Both the optical fiber fusion splicer 1 and the measurement equipment 3 are connected to an apparatus 5 for automatic off-line optimization of the splice loss estimator of the optical fiber fusion splicer 1 by means of respective connection lines 7, 9.The apparatus 5 is typically a computer.","The splicer system is typically implemented as an off-line PC and/or a PC-platform based splicer.","The platform provides the reliable communication among the computer, the splicer and the measurement instruments.","It supports the fast data/parameters transition and automatic parameter optimization, and it also opens the possibility for development of application programs like off-line tuning.","In order to make an automatic optimization of the splice loss-estimator a splice database is established in the apparatus 5, where the splice database includes results obtained by the estimator from a number of splices (i.e.","estimated splice losses and information derived from the at least one image taken of the respective splice), the estimation parameters used in the splice loss estimation procedures, as well as the corresponding measured splice losses.","A spreadsheet-based software, e.g.","Microsoft® Excel, is one of the most commonly used commercial software packages for spreadsheet applications (e.g.","data collection, sorting and analysis etc.).","In the present invention, Microsoft® Excel is used as a basic software platform to form most application software modules for off-line tuning (see FIG.","1).","These modules include a module 11 for configuration of measurement instruments, modules 13, 15, 17 for uploading and downloading data/parameters, a module 19 for data collection and sorting, and a module 21 for off-line tuning.","The arrows in FIG.","1 show the paths of data flow.","Image-Derived Information Data and Estimation Parameters The splice loss database for off-line tuning is established based essentially on two major parts: (1) data of the measured splice losses obtained from measurement instruments, i.e.","measured splice losses; and (2) information data derived from the at least one image taken of the respective splice, and estimation parameters obtained from the estimator.","Uploading the data of the measured splice losses is straightforward.","The data of splice losses obtained from measurement instrument 3 (e.g.","a Hewlett Packard 8153A Lightwave Multimeter) for each splice can be directly uploaded into the Excel worksheet using standard communication methods, e.g.","by means of a general purpose interface bus (GPIB), serial communication or direct data acquisition as shown in FIG.","1.On the other hand, however, the image-derived information data and the estimation parameters obtained from the estimator have to be carefully selected since the models for splice loss estimation depends on the fusion processes and the fiber types being used.","For clarity, the procedures for construction of the splice database and for optimization of the splice loss estimator of an Ericsson FSU995 Fusion Splicer are described below as an illustrative example.","The causes of splice losses in the Ericsson FSU995 splicer have been classified into four portions according to different loss mechanisms, see FIG.","2.Splice loss LM due to core deformation (the micro bending loss LD when the MFD mismatch is taken into account).","This loss is mainly caused by bad fiber quality (e.g.","high cladding to core eccentricity, high non-circularity of the fiber etc.","), by bad cleave angles, by high cladding offset after alignment, and by inappropriate selection of fusion processes and/or fusion parameters.","Splice loss LI due to mismatch of hot core index.","This loss originates from directional diffusion processes caused by significant difference of the dopant concentrations in the fibers, and significant difference in the development of fiber core deformation due to different fiber core diameters.","Splice loss LA due to macro bending.","This loss is mainly caused by imperfect longitude alignment of the fibers because of an inappropriate force applied on the stripped surfaces of the fibers, and microdust of large size remaining on the fiber surfaces.","Splice loss shift LS.","This loss is caused by the systematic errors due to systematic errors in the measurement system and imperfections of mechanical system in the splicer.","It usually presents as a constant shift in the splice losses.","The total splice loss LT can then be expressed by summation of the losses from the different mechanisms: LT=LM+LI+LA+LS (1) These losses can be expressed as functions of the estimation parameters and the analyzed results (i.e.","image-originating parameters) derived by the estimator.","i LM=LM{LD, F factor(MFDLL, MFDR), M factor(MFDL, MFDR)} (2) LI=LI{V, loss factor, Index limit} (3) LA=LA{view angle offset, macro factor, MFDL, MFDR} (4) Here, the analyzed results (image-derived parameters) used in the estimation are the micro bending loss LD and the hot core index offset V obtained from hot images, and the view angle offset (i.e.","the view angle difference between left and right fibers) obtained from cold images.","The estimation parameters are the M and F factors (which are uniquely determined by means of the MFDL and MFDR), the loss factor, the index limit, the macro factor for adjustment of LA, the MFDL and MFDR representing MFD of left and right fibers, as well as LS.","The loss factor and the index limit are parameters designed for erbium fiber splicing processes only.","The basic procedures how to derive the total loss from primary information of the images and the estimation parameters is schematically illustrated in the FIG.","2.Off-Line Tuning Procedures For a sequence of n splices, a splice database for the off-line tuning can be created by use of the upload software modules.","Assuming that the measured splice loss for the i'th splice is LMiε{LM1LM2, .",".",".","LMn}, i=-1, 2, .",".",".","n, and the estimated splice loss corresponding to the i'th splice is LTiε{LT1, LT2, .",".",".","LTn} the splice database will thus be: Database={LMi, LTi(Pj, Cik)}, i=1, 2, .",".",".","n; j=1, 2, .",".",".","; k=1, 2, .",".",".","(5) where, Pj and Cik are the estimation parameters and the analyzed results obtained from the estimator, respectively.","Note particularly that LMi is the measured loss for splice i and shall not be mixed up with the splice loss LM due to core deformation.","Further, the estimation parameters Pj, j=1, 2, .",".",".",", are different for different fusion processes used and for different kind of fibers spliced.","After uploading all data into the worksheet, it is important to sort the data according to fusion processes being used.","There are two reasons for data sort.","(1) Some parameters have been designed not only for optimizing the estimator but also for controlling the fusion processes.","These parameters should not be optimized during the off-line tuning if they have been used as control parameters in the fusion processes.","This is because any optimization of these parameters might change the fusion process completely.","The typical parameter in Ericsson FSU995 splicers is the index limit.","(2) The models used for different fusion processes in the estimator might be completely different.","For instance, in a normal splice, the contributions from the micro bending loss LD dominate the total splice losses.","Therefore, the model calculations are based mainly on the LD.","On the other hand, however, if the core offset of two fibers is large, e.g.",">1 μm, the splice losses will strongly depend not only on LD but also on the wavelength of the signal being transmitted.","In this case, a new model for wavelength dependence losses has to be used for fully description of loss mechanisms.","The typical example in Ericsson FSU 995 splicers is the estimation processes for the attenuator maker.","The Bellcore Criteria for Optimization The basic objective of the off-line tuning is to optimize the estimation parameters so that any, or preferably the best, set of parameters fulfilled the Bellcore splice loss estimator accuracy criteria can be found.","The Bellcore accuracy criteria is defined for 90% and 100% of splice populations, respectively, in Table 1: TABLE 1 Bellcore Accuracy Criteria for Splice loss estimator Estimating range Requirement Objective Actual loss 90% 100% 90% 100% ≦0.40 dB ±0.10 dB ±0.25 dB ±0.05 dB ±0.20 dB >0.40 dB ±25% ±50% ±15% ±30% Based on the Bellcore criteria, the algorithms for evaluation of loss estimation can be derived.","As an example, the Bellcore criteria used for the off-line tuning in Ericsson FSU995 splicer is given below: Li,min(LMi)≦LTi(P1, P2, P3, P4, P5, P6, Ci1, Ci2, Ci3)≦Li,max(LMi); i=1, 2, .",".",".",", n (6) where P1, P2, P3, P4, P5, P6 are the estimation parameters (i.e.","MFDL, MFDR, loss factor, index limit, macro factor and the LS) for a given fusion process and given kind of fiber.","Similarly, Ci1, Ci2, Ci3 are the analyzed results (i.e.","LD, V, and the view angle offset).","For the i'th splice, Cik (k=1,2,3) are given (i.e.","derived from hot an cold images, respectively), whereas LTi can be varied by selecting suitable set of Pj (j=1,2 .",".",".","6).","The tunable range of LTi can be controlled by criteria parameters Li,max and Li,min(LMi).","For simplicity, we assume, here, that the maximum splice loss is 2.0 dB.","Thus, the criteria for the i'th'splice are given by the following relations.","For 90% of splice population: Li,maxLMi)=LMi+0.10 if Lmi≦0.4 dB LMi+0.25 LMI if 0.4<LMI≦2.0 dB (7) Li,min(LMi)=LMi−0.10 if LMI≦0.4 dB LMi−0.25 LMi if 0.4<LMi≦2.0 dB (8) For 100% of splice population: Li,max(LMi+0.25 if LMi≦0.4 dB LMi+0.50 LMi if 0.4<LMi≦2.0 dB (9) Li,min(LMi)=LMi−0.25 if LMi≦0.4 dB lMi−0.50 LMi if 0.4<LMi≦2.0 dB (10) In principle, the estimation parameters could be optimized by continuously varying all parameters Pj with well-defined optimization routines and simultaneously calculating LTi with formulas (1)-(4) until the Bellcore criteria, i.e.","formula (6), is fulfilled for all splices.","FIG.","3 shows a typical example of the optimized data that fulfills the Bellcore requirement.","The areas within dashed lines and solid lines, respectively, represent the 90% and the 100% Bellcore criteria, respectively.","In this particular case, both 90% and 100% Bellcore criteria are completely fulfilled.","Regression Analysis Criteria for the Best Solution It can be noticed that a set of optimized parameters that fulfill the Bellcore criteria is not unique.","Different set of optimized parameters that fulfill the Bellcore criteria may be quite different, depending on optimization routines and parameter combinations being used for optimization.","Though, even if any solution that fulfills the Bellcore criteria should be considered as an acceptable solution, it is still an open question whether it is interesting to select the best solution within Bellcore criteria.","The best solution could be defined as the set of parameters that maximize the probability that the estimated slice loss data represent the measured splice loss data, i.e.","the data distribution in FIG.","3 should be optimized to be as closed to the diagonal line (dash-doted line in FIG.","3) as possible.","In order to search for the best solution, the method of regression analysis is used.","Using such method, a regression line is generated by the “least-square fitting method” to give the best fit for the data set {LMi, LTi(Pj,Cik)}.","The regression line can be expressed by: LT=aLM+b (11) where: a = n ( ∑ i = 1 n L Mi L Ti ) - ( ∑ i = 1 n L Mi ) ( ∑ i = 1 n L Ti ) n { ∑ i = 1 n ( L Mi 2 ) } - ( ∑ i = 1 n L Mi ) 2 and ( 12 ) b = ( ∑ i = 1 n L Ti ) { ∑ i = 1 n ( L Mi 2 ) } - ( ∑ i = 1 n L Mi ) ( ∑ i = 1 n L Mi L Ti ) n { ∑ i = 1 n ( L Mi 2 ) } - ( ∑ i = 1 n L Mi ) 2 ( 13 ) Thus, the criteria for the best solution is the data set that not only fulfill the Bellcore criteria but the regression analysis criteria as well.","The regression analysis criteria is given by |a−1|≦ε1 |b|≦ε2 (14) where, a and b are the slope and the interception of the regression line.","ε1 and ε2 are the infinitesimal constants that set the conditions for controlling and terminating the optimization procedures.","In practice, however, additional criteria have to be taken into consideration, e.g.","computing time restraints.","Since many factors (e.g.","hot spots, arc sparks) can not be completely handled by the estimator, it might not be possible to optimize the estimation parameters so that the data set completely fulfill the Bellcore criteria within finite computing time.","Therefore, the criteria with well-defined computing loops and/or computing time have to be employed to terminate optimization processes if necessary.","In FIGS.","4-6, the concept of the above-described two-step optimization is demonstrated.","FIG.","4 shows the original data set that are uploaded without any optimization.","FIG.","5 shows that the first step of optimization that fulfills the Bellcore criteria.","In the second step of optimization, shown in FIG.","6, the best solution is found by use of regression analysis criteria.","Program Flowing Chart Based on the basic concepts and techniques discussed above, a simplified program flowing chart used for the off-line tuning in the Ericsson FSU995 splicer is given in FIG.","7.A splice database {LMi, LTi(Pj, Cik)} is in a step 71 (which may indeed include a plurality of sub-steps) imported into the offline tuning model 21 of the apparatus 5 as shown in FIG.","1.Then, in a step 73, the estimation parameters Pj are altered and W is calculated by using formulas (1)-(4).","In a step 75 it is checked whether LTi fulfills formula (6) and if not formula (6) is fulfilled the algorithm is returned to step 73.If formula (6) is fulfilled, then in a step 77 a regression line LT=aLM+b is generated.","Then, in a step 79, it is checked whether formula 14 is fulfilled and if not formula (14) is fulfilled the algorithm is returned to step 73.If formula (14) is fulfilled, the estimation parameters are exported, in a step 81, to the download module 17 of FIG.","1, whereafter the algorithm is ended.","Optionally, in a step 83 a splice loss diagram is created similar to those shown in FIGS.","3 and 5-6.It will be obvious that the invention may be varied in a plurality of ways.","Such variations are not to be regarded as a departure from the scope of the invention.","All such modifications as would be obvious to one skilled in the art are intended to be included within the scope of the appended claims."]],"string":"[\n [\n \"Automatic optimization of a splice loss estimator for optical fiber splicers\",\n \"A technique is provided for automatic optimization of a splice loss estimator of a fiber splicer (1), where the splice loss estimator is adapted, in a splice loss estimation procedure, to estimate the splice losses (Lti) of splices (i) of optical fibers as produced by the fiber splicer from images taken of the optical fibers at the splicing thereof, and the splice loss estimation procedure includes the use of splice loss estimation parameters (Pj).\",\n \"The estimator estimates splice losses based on information (Cij) obtained from the images and the estimation parameters.\",\n \"Further, the splice losses are measured by means of a measurement instrument (3).\",\n \"The estimated (Lti) and measured (LMi) splice losses, and the information obtained from the images are uploaded (71) into an off-line computer (5) and the key estimation parameters are automatically optimized by the selection of any solution within the Bellcore accuracy criteria (75), whereafter the optimized estimation parameters are downloaded (81) to the splicer.\"\n ],\n [\n \"1.An apparatus for automatic optimization of a splice loss estimator of an optical fiber splicer apparatus, said splice loss estimator being adapted, in a splice loss estimation procedure, to estimate the splice losses (LTi) of splices (i) of end portions of optical fibers as produced by said fiber splicer apparatus from at least one respective image taken of the respective end portions of the optical fibers prior to, during, or subsequent to the splicing thereof, said splice loss estimation procedure including the use of a set of splice loss estimation parameters (Pj), comprising a first input line for receiving, from said splice loss estimator, data of a plurality of splices of end portions of optical fibers as produced by said fiber splicer apparatus, wherein said data for each splice (i) include information (Cij) derived from the at least one image taken of the end portions of the optical fibers constituting that splice, and the estimated splice loss (LTi) of that splice; a second input line for receiving a splice loss value (LMi) for each of said plurality of splices of end portions of optical fibers as measured by a measurement equipment; means for determining a new set of splice loss estimation parameters from said data and said estimated and measured splice loss values such that said splice loss estimation procedure, using the new set of splice loss estimation parameters, estimates the splice loss of each of said plurality of splices of end portions of optical fibers with a given accuracy; and an output line for outputting said determined new set of splice loss estimation parameters to said splice loss estimator to replace the set of splice loss estimation parameters used in the splice loss estimation procedure of the splice loss estimator for the estimation of the splice losses of any further splices of end portions of optical fibers to be produced by said fiber splicer apparatus.\",\n \"2.The apparatus as claimed in claim 1 wherein said data for each splice (i) include information (Cij) derived from hot images taken of the end portions of the optical fibers constituting that splice.\",\n \"3.The apparatus as claimed in claim 2 wherein said data for each splice include information regarding mismatch of mode field diameter (MFD), including micro bending loss (LD), or fiber core/cladding deformation including cladding-to-core eccentricities, non-circularity of fibers, cleave angle of fiber ends, and/or cladding/core offsets.\",\n \"4.The apparatus as claimed in claim 2 wherein said data for each splice include information regarding mismatch of hot core index (LI), including hot core index offset (V), fiber dopant concentrations, and/or fiber core diameters.\",\n \"5.The apparatus as claimed in claim 1 wherein said data for each splice (i) include information (Cij) derived from cold images taken of the end portions of the optical fibers constituting that splice.\",\n \"6.The apparatus as claimed in claim 5 wherein said data for each splice include information regarding macro bending, including view angle offset, fiber alignment, and/or presence of microdust on fiber surfaces.\",\n \"7.The apparatus as claimed in claim 1 wherein said apparatus is a computer, preferably an off-line computer and/or a PC-platform based splicer, provided with suitable software and information as regards the splice loss estimation procedure used in the splice loss estimator.\",\n \"8.The apparatus as claimed in claim 1 wherein said set of splice loss estimation parameters include any of an M factor, an F factor, a loss factor, a macro factor, a mode field diameter left (MFDL) factor, a mode field diameter right (MFDR) factor, and a splice loss shift (LS) factor, where the M and F factors are employed in the estimation of a core deformation induced splice loss (LM), the loss factor is employed in the estimation of a hot core index mismatch induced splice loss (LI) the macro factor, the mode field diameter left (MFDL) and right (MFDR) factors are employed in the estimation of a macro bending induced splice loss (LA) and the splice loss shift (LS) factor is employed in the estimation of a splice loss correction term, which corrects for any systematic errors in the estimation procedure.\",\n \"9.The apparatus as claimed in claim 1 wherein said apparatus is adapted to receive, on said first input line, the set of estimation parameters as used by said splicer apparatus.\",\n \"10.The apparatus as claimed in claim 1 wherein said apparatus is adapted to receive, on said first input line, a set of image processing parameters, wherein said set of image processing parameters are used in said splice loss estimation procedure by said splice loss estimator of the splicer apparatus; said means for determining is adapted to determine a new set of image processing parameters from said data and said estimated and measured splice loss values such that said splice loss estimation procedure, using the new set of image processing parameters, estimates the splice loss of each of said plurality of splices of end portions of optical fibers with a given accuracy; and said apparatus for automatic optimization is adapted to output, via said output line, said determined new set of image processing parameters to said splice estimator to replace the set of image processing parameters used in the splice loss estimation procedure of the splice estimator for the estimation of the splice losses of any further splices of end portions of optical fibers to be produced by said fiber splicer apparatus.\",\n \"11.The apparatus as claimed in claim 1 wherein said plurality of splices are at least 50, more preferably at least 100, and most preferably between 100 and 500.12.The apparatus as claimed in claim 1 wherein said means for determining is adapted to determine said new set of splice loss estimation parameters such that said new set of splice loss estimation parameters fulfils the Bellcore splice loss estimator criteria, preferably the 90% Bellcore criteria, and most preferably the 100% Bellcore criteria.\",\n \"13.The apparatus as claimed in claim 1 wherein said means for determining is adapted to determine said new set of splice loss estimation parameters by using linear regression criteria to increase the probability that the estimated splice loss values using said new set of estimation parameters represent the measured splice losses.\",\n \"14.\",\n \"(canceled) 15.A method for automatic optimization of a splice loss estimator of an optical fiber splicer apparatus, where said splice loss estimator is adapted, in a splice loss estimation procedure, to estimate the splice losses (LTi) of splices (i) of end portions of optical fibers as produced by said fiber splicer apparatus from at least one respective image taken of the respective end portions of the optical fibers prior to, during, or subsequent to the splicing thereof, said splice loss estimation procedure including the use of a set of splice loss estimation parameters (Pj), comprising the steps of: receiving, from said splice loss estimator, data of a plurality of splices of end portions of optical fibers as produced by said fiber splicer apparatus, wherein said data for each splice (i) include information (Cij) derived from the at least one image taken of the end portions of the optical fibers constituting that splice, and the estimated splice loss (LTi) of that splice; receiving a splice loss value (LMi) for each of said plurality of splices of end portions of optical fibers as measured by a measurement equipment; determining a new set of splice loss estimation parameters from said data and said estimated and measured splice loss values such that said splice loss estimation procedure, using the new set of splice loss estimation parameters, estimates the splice loss of each of said plurality of splices of end portions of optical fibers with a given accuracy; and outputting said determined new set of splice loss estimation parameters to said splice loss estimator to replace the set of splice loss estimation parameters used in the splice loss estimation procedure of the splice loss estimator for the estimation of the splice losses of any further splices of end portions of optical fibers to be produced by said fiber splicer apparatus.\",\n \"16.The method as claimed in claim 15 wherein said data for each splice (i) include information (Cij) derived from hot images taken of the end portions of the optical fibers constituting that splice.\",\n \"17.The method as claimed in claim 15 wherein said data for each splice (i) include information (Cij) derived from cold images taken of the end portions of the optical fibers constituting that splice.\",\n \"18.The method as claimed in claim 15 wherein said set of splice loss estimation parameters include any of an M factor, an F factor, a loss factor, a macro factor, a mode field diameter left (MFDL) factor, a mode field diameter right (MFDR) factor, and a splice loss shift (LS) factor, where the M and F factors are employed in the estimation of a core deformation induced splice loss (LM), the loss factor is employed in the estimation of a hot core index mismatch induced splice loss (LI) the macro factor, the mode field diameter left (MFDL) and right (MFDR) factors are employed in the estimation of a macro bending induced splice loss (LA) and the splice loss shift (LS) factor is employed in the estimation of a splice loss correction term, which corrects for any systematic errors in the estimation procedure.\",\n \"19.The method as claimed in claim 15 further comprising the steps of: receiving, from said splice estimator, a set of image processing parameters, wherein said set of image processing parameters are used in said splice loss estimation procedure by said splice loss estimator of the splicer apparatus; determining a new set of image processing parameters from said data and said estimated and measured splice loss values such that said splice loss estimation procedure, using the new set of image processing parameters, estimates the splice loss of each of said plurality of splices of end portions of optical fibers with a given accuracy; and outputting said determined new set of image processing parameters to said splice estimator to replace the set of image processing parameters used in the splice loss estimation procedure of the splice estimator for the estimation of the splice losses of any further splices of end portions of optical fibers to be produced by said fiber splicer apparatus.\",\n \"20.The method as claimed in claim 15 wherein said new set of splice loss estimation parameters is determined such that said new set of splice loss estimation parameters fulfils the Bellcore splice loss estimator criteria, preferably the 90% Bellcore criteria, and most preferably the 100% Bellcore criteria.\",\n \"21.The method as claimed in claim 15 wherein said new set of splice loss estimation parameters is determined by using linear regression criteria to increase the probability that the estimated splice loss values using said new set of estimation parameters represent the measured splice losses.\",\n \"22.\",\n \"(canceled) 23.A method for automatic optimization of a splice loss estimator of an optical fiber splicer apparatus, where said splice loss estimator is adapted, in a splice loss estimation procedure, to estimate the splice losses (LTi) Of splices of end portions of optical fibers as produced by said fiber splicer apparatus from at least one respective image taken of the respective end portions of the optical fibers prior to, during, or subsequent to the splicing thereof, said splice loss estimation procedure including the use of a set of splice loss estimation parameters (Pj), comprising the steps of: producing a plurality of splices of end portions of optical fibers by means of said fiber splicer apparatus; deducing information from the at least one image taken for each splice produced; estimating the splice loss based on the information deduced from the at least one image for each splice produced; measuring the splice loss of each splice produced by means of a measurement instrument,; uploading the deduced information, the estimated splice losses and the measure splice losses to a processing device; determining a new set of splice loss estimation parameters from the deduced information and the estimated and measured splice loss values such that said splice loss estimation procedure, using the new set of splice loss estimation parameters, estimates the splice loss of each of said plurality of splices of end portions of optical fibers with a given accuracy; and downloading said determined new set of splice loss estimation parameters to said splice loss estimator of the fiber splicer apparatus to replace the set of splice loss estimation parameters used in the splice loss estimation procedure of the splice loss estimator for the estimation of the splice losses of any further splices of end portions of optical fibers to be produced by said fiber splicer apparatus.\"\n ],\n [\n \"<SOH> DESCRIPTION OF RELATED ART AND BACKGROUND OF THE INVENTION <EOH>The technology for forming low-loss optical fibers has advanced to a point where there is widespread commercial manufacturing of optical fibers.\",\n \"Most processing includes drawing an optical fiber from a previously manufactured glass boule, sometimes referred to as a fiber preform.\",\n \"Extremely long lengths of fiber can be obtained by splicing a plurality of lengths, which are obtained using current fusion splicer techniques.\",\n \"Additionally, it has become increasingly more common to splice optical fibers, which have broken, either accidentally, or during appropriate proof testing.\",\n \"For these and other applications, splicing in which the coating material is removed from end portions of two fibers, which are then fused together end to end, provides a suitable means for joining the ends of two glass fibers with an acceptably low splice loss.\",\n \"Fusion splice losses depend mainly on the mismatch of mode-field diameter (MFD) and misalignment of the cladding/core of two fibers due to either the fiber refractive index design or the inappropriate fusion processes being used.\",\n \"With the help of image techniques, information on the MFD mismatch can be obtained and analyzed by monitoring the deformation/misalignment of the cladding/core of the fibers during the fusion process, see e.g.\",\n \"EP 1 014 070.Such a monitoring system includes typically a charged-coupled device (CCD) camera equipped with an image processor.\",\n \"By use of suitable theoretical models and the information obtained from the images, an estimation method for evaluation of splice losses can be established.\",\n \"The estimation method as a passive technique for evaluation of splice losses is widely used in most automated fusion splicers of today.\",\n \"Different models for evaluation of splice loss have been explored and developed during the past two decades.\",\n \"Well-known methods for splice loss estimation include the butt-joint approximation and the mode coupling theory.\",\n \"More advanced methods, e.g.\",\n \"hot-image techniques for real-time analysis of cladding and core deformation have also been developed.\",\n \"In order to achieve the best performance and maximize the flexibility of the estimation method, the models used for splice loss estimation include usually a number of free parameters, called estimation parameters.\",\n \"The optimization of these estimation parameters in order to achieve the best performance of the estimation method has nowadays become one of essential features in the development of splicing techniques.\",\n \"In practice, the estimation parameters are manually optimized according to different types of on-test fibers and the fusion processes being used.\",\n \"Due to primarily technical reasons and rather complicated nature of the involved splicing processes, the optimization of the estimation parameters is a quite difficult and time consuming job that may only be performed by very experienced engineers.\",\n \"The time typically needed for manually optimizing the estimation parameters on a given fiber combination is as long as a few days which is hardly to be accepted by splicer users, especially, when frequently changes of fusion processes and fiber combinations are involved, e.g.\",\n \"in the manufacture of erbium doped fiber amplifiers (EDFA).\",\n \"This is hardly acceptable for the splicer user, and such optimization has thus typically to be performed by the fiber splicer manufacturer.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>The present inventors have realized that the splicer user does not need to re-optimize the image processing parameters, but only the estimation parameters.\",\n \"For a fiber splicer user, only a limited number of the image processing parameters can be optimized, and most of these parameters are already designed to be linear to the splice loss by an embedded algorithm in the splicer.\",\n \"In the present invention a new technique for fast automatic optimization of the performance of the estimator is proposed.\",\n \"The technique leaves the image processing of the cold and hot images taken during the fusion processes unchanged.\",\n \"The built-in estimator in the splicer analyzes the causes of splice losses based on the information obtained from the images, and based on the fusion process employed and the kind of spliced fibers.\",\n \"Further, splice losses are measured by means of a measurement instrument, e.g.\",\n \"a power meter.\",\n \"The results of the analysis and the measured splice losses are uploaded into a spreadsheet-based worksheet in an off-line computer and/or a PC-platform based splicer to form a splice database.\",\n \"Then, the manual procedures for optimization of estimation parameters are simulated.\",\n \"The key estimation parameters are automatically optimized by the selection of any solution within the Bellcore accuracy criteria, or the best solution within the Bellcore accuracy criteria by assistance of regression analysis criteria, whereafter the optimized estimation parameters are downloaded to splicer.\",\n \"The time needed for optimizing a splice database with hundred splices is significantly reduced down to a few seconds only.\",\n \"Thus, an enhanced performance of the estimator is automatically achieved.\",\n \"The tedious and complicated work for manually optimizing the estimator is automated.\",\n \"It is a main object of the present invention to provide an apparatus for automatic optimization of a splice loss estimator of an optical fiber splicer, wherein the splice loss estimator is adapted, in a splice loss estimation procedure, to estimate the splice losses of splices of end portions of optical fibers as produced by said fiber splicer apparatus from at least one respective image taken of the respective end portions of the optical fibers prior to, during, or subsequent to the splicing thereof, and wherein the splice loss estimation procedure includes the use of a set of splice loss estimation parameters, which apparatus for automatic optimization is fast, and which is particularly suited to be used by a fiber splicer user.\",\n \"It is in this respect a particular object of the invention to provide such apparatus, which is simple, easy to use, accurate, precise and of low cost.\",\n \"These objects, among others, are according to a first aspect of the present invention attained by an apparatus comprising a first input line; a second input line; means for determining (preferably a personal computer provided with suitable software); and an output line.\",\n \"The first input line is provided for receiving, from the splice loss estimator, data of a plurality of splices of end portions of optical fibers as produced by the fiber splicer apparatus, wherein the data for each splice include information derived from the image taken of the end portions of the optical fibers constituting that splice, and the estimated splice loss of that splice; and the second input line is provided for receiving a splice loss value for each of the plurality of splices of end portions of optical fibers as measured by a measurement equipment.\",\n \"The means for determining is adapted to determine or calculate a new set of splice loss estimation parameters from the data and the measured splice loss values such that the splice loss estimation procedure, using the new set of splice loss estimation parameters, estimates the splice loss of each of the plurality of splices of end portions of optical fibers with a given accuracy.\",\n \"Finally, the output line is provided for outputting the determined or calculated new set of splice loss estimation parameters to the splice loss estimator of the fiber splicer apparatus to replace the set of splice loss estimation parameters used in the splice loss estimation procedure of the splice loss estimator for the estimation of the splice losses of any further splices of end portions of optical fibers to be produced by the fiber splicer apparatus.\",\n \"Further, the above-mentioned objects, among others, are according to a second aspect of the present invention attained by an optical fiber splicer system comprising such an apparatus for automatic optimization.\",\n \"It is a further object of the present invention to provide a method for automatic optimization of the above-depicted splice loss estimator, which is fast and which is thus suitable to be used by the fiber splicer user.\",\n \"This object is according to a third aspect of the present invention attained by a method comprising the following steps: (i) receiving, from said splice loss estimator, data of a plurality of splices of end portions of optical fibers as produced by said fiber splicer apparatus, wherein said data for each splice include information derived from the at least one image taken of the end portions of the optical fibers constituting that splice, and the estimated splice loss of that splice; (ii) receiving a splice loss value for each of said plurality of splices of end portions of optical fibers as measured by a measurement equipment; (iii) determining a new set of splice loss estimation parameters from said data and said measured splice loss values such that said splice loss estimation procedure, using the new set of splice loss estimation parameters, estimates the splice loss of each of said plurality of splices of end portions of optical fibers with a given accuracy; and (iv) outputting said determined new set of splice loss estimation parameters to said splice loss estimator to replace the set of splice loss estimation parameters used in the splice loss estimation procedure of the splice loss estimator for the estimation of the splice losses of any further splices of end portions of optical fibers to be produced by said fiber splicer apparatus.\",\n \"Finally, according to a fourth aspect of the present invention there is provided a computer program product loadable into the internal memory of a computer, which comprises software code portions for performing the above-described method when the computer program product is run on the computer.\",\n \"Further characteristics of the invention and advantages thereof will be evident from the detailed description of preferred embodiments of the invention given hereinbelow and the accompanying drawings, which are given by way of illustration only, and thus are not limitative of the present invention.\"\n ],\n [\n \"TECHNICAL FIELD OF THE INVENTION The present invention generally relates to fiber optics.\",\n \"More specifically, the invention relates to -an apparatus and a method, respectively, for automatic optimization of a splice loss estimator for optical fiber splicers, to an optical fiber splicer system comprising such an apparatus, and to a computer program product for performing such a method.\",\n \"DESCRIPTION OF RELATED ART AND BACKGROUND OF THE INVENTION The technology for forming low-loss optical fibers has advanced to a point where there is widespread commercial manufacturing of optical fibers.\",\n \"Most processing includes drawing an optical fiber from a previously manufactured glass boule, sometimes referred to as a fiber preform.\",\n \"Extremely long lengths of fiber can be obtained by splicing a plurality of lengths, which are obtained using current fusion splicer techniques.\",\n \"Additionally, it has become increasingly more common to splice optical fibers, which have broken, either accidentally, or during appropriate proof testing.\",\n \"For these and other applications, splicing in which the coating material is removed from end portions of two fibers, which are then fused together end to end, provides a suitable means for joining the ends of two glass fibers with an acceptably low splice loss.\",\n \"Fusion splice losses depend mainly on the mismatch of mode-field diameter (MFD) and misalignment of the cladding/core of two fibers due to either the fiber refractive index design or the inappropriate fusion processes being used.\",\n \"With the help of image techniques, information on the MFD mismatch can be obtained and analyzed by monitoring the deformation/misalignment of the cladding/core of the fibers during the fusion process, see e.g.\",\n \"EP 1 014 070.Such a monitoring system includes typically a charged-coupled device (CCD) camera equipped with an image processor.\",\n \"By use of suitable theoretical models and the information obtained from the images, an estimation method for evaluation of splice losses can be established.\",\n \"The estimation method as a passive technique for evaluation of splice losses is widely used in most automated fusion splicers of today.\",\n \"Different models for evaluation of splice loss have been explored and developed during the past two decades.\",\n \"Well-known methods for splice loss estimation include the butt-joint approximation and the mode coupling theory.\",\n \"More advanced methods, e.g.\",\n \"hot-image techniques for real-time analysis of cladding and core deformation have also been developed.\",\n \"In order to achieve the best performance and maximize the flexibility of the estimation method, the models used for splice loss estimation include usually a number of free parameters, called estimation parameters.\",\n \"The optimization of these estimation parameters in order to achieve the best performance of the estimation method has nowadays become one of essential features in the development of splicing techniques.\",\n \"In practice, the estimation parameters are manually optimized according to different types of on-test fibers and the fusion processes being used.\",\n \"Due to primarily technical reasons and rather complicated nature of the involved splicing processes, the optimization of the estimation parameters is a quite difficult and time consuming job that may only be performed by very experienced engineers.\",\n \"The time typically needed for manually optimizing the estimation parameters on a given fiber combination is as long as a few days which is hardly to be accepted by splicer users, especially, when frequently changes of fusion processes and fiber combinations are involved, e.g.\",\n \"in the manufacture of erbium doped fiber amplifiers (EDFA).\",\n \"This is hardly acceptable for the splicer user, and such optimization has thus typically to be performed by the fiber splicer manufacturer.\",\n \"SUMMARY OF THE INVENTION The present inventors have realized that the splicer user does not need to re-optimize the image processing parameters, but only the estimation parameters.\",\n \"For a fiber splicer user, only a limited number of the image processing parameters can be optimized, and most of these parameters are already designed to be linear to the splice loss by an embedded algorithm in the splicer.\",\n \"In the present invention a new technique for fast automatic optimization of the performance of the estimator is proposed.\",\n \"The technique leaves the image processing of the cold and hot images taken during the fusion processes unchanged.\",\n \"The built-in estimator in the splicer analyzes the causes of splice losses based on the information obtained from the images, and based on the fusion process employed and the kind of spliced fibers.\",\n \"Further, splice losses are measured by means of a measurement instrument, e.g.\",\n \"a power meter.\",\n \"The results of the analysis and the measured splice losses are uploaded into a spreadsheet-based worksheet in an off-line computer and/or a PC-platform based splicer to form a splice database.\",\n \"Then, the manual procedures for optimization of estimation parameters are simulated.\",\n \"The key estimation parameters are automatically optimized by the selection of any solution within the Bellcore accuracy criteria, or the best solution within the Bellcore accuracy criteria by assistance of regression analysis criteria, whereafter the optimized estimation parameters are downloaded to splicer.\",\n \"The time needed for optimizing a splice database with hundred splices is significantly reduced down to a few seconds only.\",\n \"Thus, an enhanced performance of the estimator is automatically achieved.\",\n \"The tedious and complicated work for manually optimizing the estimator is automated.\",\n \"It is a main object of the present invention to provide an apparatus for automatic optimization of a splice loss estimator of an optical fiber splicer, wherein the splice loss estimator is adapted, in a splice loss estimation procedure, to estimate the splice losses of splices of end portions of optical fibers as produced by said fiber splicer apparatus from at least one respective image taken of the respective end portions of the optical fibers prior to, during, or subsequent to the splicing thereof, and wherein the splice loss estimation procedure includes the use of a set of splice loss estimation parameters, which apparatus for automatic optimization is fast, and which is particularly suited to be used by a fiber splicer user.\",\n \"It is in this respect a particular object of the invention to provide such apparatus, which is simple, easy to use, accurate, precise and of low cost.\",\n \"These objects, among others, are according to a first aspect of the present invention attained by an apparatus comprising a first input line; a second input line; means for determining (preferably a personal computer provided with suitable software); and an output line.\",\n \"The first input line is provided for receiving, from the splice loss estimator, data of a plurality of splices of end portions of optical fibers as produced by the fiber splicer apparatus, wherein the data for each splice include information derived from the image taken of the end portions of the optical fibers constituting that splice, and the estimated splice loss of that splice; and the second input line is provided for receiving a splice loss value for each of the plurality of splices of end portions of optical fibers as measured by a measurement equipment.\",\n \"The means for determining is adapted to determine or calculate a new set of splice loss estimation parameters from the data and the measured splice loss values such that the splice loss estimation procedure, using the new set of splice loss estimation parameters, estimates the splice loss of each of the plurality of splices of end portions of optical fibers with a given accuracy.\",\n \"Finally, the output line is provided for outputting the determined or calculated new set of splice loss estimation parameters to the splice loss estimator of the fiber splicer apparatus to replace the set of splice loss estimation parameters used in the splice loss estimation procedure of the splice loss estimator for the estimation of the splice losses of any further splices of end portions of optical fibers to be produced by the fiber splicer apparatus.\",\n \"Further, the above-mentioned objects, among others, are according to a second aspect of the present invention attained by an optical fiber splicer system comprising such an apparatus for automatic optimization.\",\n \"It is a further object of the present invention to provide a method for automatic optimization of the above-depicted splice loss estimator, which is fast and which is thus suitable to be used by the fiber splicer user.\",\n \"This object is according to a third aspect of the present invention attained by a method comprising the following steps: (i) receiving, from said splice loss estimator, data of a plurality of splices of end portions of optical fibers as produced by said fiber splicer apparatus, wherein said data for each splice include information derived from the at least one image taken of the end portions of the optical fibers constituting that splice, and the estimated splice loss of that splice; (ii) receiving a splice loss value for each of said plurality of splices of end portions of optical fibers as measured by a measurement equipment; (iii) determining a new set of splice loss estimation parameters from said data and said measured splice loss values such that said splice loss estimation procedure, using the new set of splice loss estimation parameters, estimates the splice loss of each of said plurality of splices of end portions of optical fibers with a given accuracy; and (iv) outputting said determined new set of splice loss estimation parameters to said splice loss estimator to replace the set of splice loss estimation parameters used in the splice loss estimation procedure of the splice loss estimator for the estimation of the splice losses of any further splices of end portions of optical fibers to be produced by said fiber splicer apparatus.\",\n \"Finally, according to a fourth aspect of the present invention there is provided a computer program product loadable into the internal memory of a computer, which comprises software code portions for performing the above-described method when the computer program product is run on the computer.\",\n \"Further characteristics of the invention and advantages thereof will be evident from the detailed description of preferred embodiments of the invention given hereinbelow and the accompanying drawings, which are given by way of illustration only, and thus are not limitative of the present invention.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS FIG.\",\n \"1 illustrates, schematically, in a block diagram, an optical fiber splicer system comprising an optical fiber fusion splicer, measurement instruments, and an apparatus for automatic off-line optimization of a splice loss estimator for the optical fiber fusion splicer according to a general embodiment of the present invention.\",\n \"FIG.\",\n \"2 illustrates schematically procedures for deriving splice losses.\",\n \"FIG.\",\n \"3 illustrates the result of a splice loss estimation procedure, where the Bellcore criteria are fulfilled, which estimation procedure uses a new set of splice loss estimation parameters resulting from the automatic optimization according to the present invention.\",\n \"FIGS.\",\n \"4-6 illustrate the results of splice loss estimation procedures using the original set of estimation parameters (FIG.\",\n \"4); and new sets of splice loss estimation parameters resulting from the automatic optimization according to the present invention (FIGS.\",\n \"5-6), respectively.\",\n \"FIG.\",\n \"7 is a schematic flow chart illustrating an off-line tuning procedure according to a preferred embodiment of the present invention.\",\n \"DETAILED DESCRIPTION OF EMBODIMENTS In the following description, for purposes of explanation and not limitation, specific details are set forth, such as particular techniques and applications in order to provide a thorough understanding of the present invention.\",\n \"However, it will be apparent to one skilled in the art that the present invention may be practiced in other embodiments that depart from these specific details.\",\n \"Optical Fiber Splicer System Including a PC-Based Platform for Supporting Off-Line Tuning With reference to FIG.\",\n \"1, which schematically illustrates an optical fiber splicer system, a general embodiment of the present invention will be overviewed.\",\n \"The system comprises an optical fiber fusion splicer 1 including a splice loss estimator (not explicitly illustrated in FIG.\",\n \"1) for, in a splice loss estimation procedure, estimating the splice losses of splices of end portions of optical fibers as produced by the fiber splicer 1 from at least one respective image taken of the respective end portions of the optical fibers prior to, during, or subsequent to the splicing thereof, where the splice loss estimation procedure includes the use of a set of splice loss estimation parameters.\",\n \"The optical fiber fusion splicer may e.g.\",\n \"be an Ericsson 995 Fusion Splicer.\",\n \"Further, there is provided measurement equipment 3 for measuring the splice loss for a number of splices produced.\",\n \"The measurement equipment may be a power meter, an extinction ratio meter, or similar.\",\n \"Both the optical fiber fusion splicer 1 and the measurement equipment 3 are connected to an apparatus 5 for automatic off-line optimization of the splice loss estimator of the optical fiber fusion splicer 1 by means of respective connection lines 7, 9.The apparatus 5 is typically a computer.\",\n \"The splicer system is typically implemented as an off-line PC and/or a PC-platform based splicer.\",\n \"The platform provides the reliable communication among the computer, the splicer and the measurement instruments.\",\n \"It supports the fast data/parameters transition and automatic parameter optimization, and it also opens the possibility for development of application programs like off-line tuning.\",\n \"In order to make an automatic optimization of the splice loss-estimator a splice database is established in the apparatus 5, where the splice database includes results obtained by the estimator from a number of splices (i.e.\",\n \"estimated splice losses and information derived from the at least one image taken of the respective splice), the estimation parameters used in the splice loss estimation procedures, as well as the corresponding measured splice losses.\",\n \"A spreadsheet-based software, e.g.\",\n \"Microsoft® Excel, is one of the most commonly used commercial software packages for spreadsheet applications (e.g.\",\n \"data collection, sorting and analysis etc.).\",\n \"In the present invention, Microsoft® Excel is used as a basic software platform to form most application software modules for off-line tuning (see FIG.\",\n \"1).\",\n \"These modules include a module 11 for configuration of measurement instruments, modules 13, 15, 17 for uploading and downloading data/parameters, a module 19 for data collection and sorting, and a module 21 for off-line tuning.\",\n \"The arrows in FIG.\",\n \"1 show the paths of data flow.\",\n \"Image-Derived Information Data and Estimation Parameters The splice loss database for off-line tuning is established based essentially on two major parts: (1) data of the measured splice losses obtained from measurement instruments, i.e.\",\n \"measured splice losses; and (2) information data derived from the at least one image taken of the respective splice, and estimation parameters obtained from the estimator.\",\n \"Uploading the data of the measured splice losses is straightforward.\",\n \"The data of splice losses obtained from measurement instrument 3 (e.g.\",\n \"a Hewlett Packard 8153A Lightwave Multimeter) for each splice can be directly uploaded into the Excel worksheet using standard communication methods, e.g.\",\n \"by means of a general purpose interface bus (GPIB), serial communication or direct data acquisition as shown in FIG.\",\n \"1.On the other hand, however, the image-derived information data and the estimation parameters obtained from the estimator have to be carefully selected since the models for splice loss estimation depends on the fusion processes and the fiber types being used.\",\n \"For clarity, the procedures for construction of the splice database and for optimization of the splice loss estimator of an Ericsson FSU995 Fusion Splicer are described below as an illustrative example.\",\n \"The causes of splice losses in the Ericsson FSU995 splicer have been classified into four portions according to different loss mechanisms, see FIG.\",\n \"2.Splice loss LM due to core deformation (the micro bending loss LD when the MFD mismatch is taken into account).\",\n \"This loss is mainly caused by bad fiber quality (e.g.\",\n \"high cladding to core eccentricity, high non-circularity of the fiber etc.\",\n \"), by bad cleave angles, by high cladding offset after alignment, and by inappropriate selection of fusion processes and/or fusion parameters.\",\n \"Splice loss LI due to mismatch of hot core index.\",\n \"This loss originates from directional diffusion processes caused by significant difference of the dopant concentrations in the fibers, and significant difference in the development of fiber core deformation due to different fiber core diameters.\",\n \"Splice loss LA due to macro bending.\",\n \"This loss is mainly caused by imperfect longitude alignment of the fibers because of an inappropriate force applied on the stripped surfaces of the fibers, and microdust of large size remaining on the fiber surfaces.\",\n \"Splice loss shift LS.\",\n \"This loss is caused by the systematic errors due to systematic errors in the measurement system and imperfections of mechanical system in the splicer.\",\n \"It usually presents as a constant shift in the splice losses.\",\n \"The total splice loss LT can then be expressed by summation of the losses from the different mechanisms: LT=LM+LI+LA+LS (1) These losses can be expressed as functions of the estimation parameters and the analyzed results (i.e.\",\n \"image-originating parameters) derived by the estimator.\",\n \"i LM=LM{LD, F factor(MFDLL, MFDR), M factor(MFDL, MFDR)} (2) LI=LI{V, loss factor, Index limit} (3) LA=LA{view angle offset, macro factor, MFDL, MFDR} (4) Here, the analyzed results (image-derived parameters) used in the estimation are the micro bending loss LD and the hot core index offset V obtained from hot images, and the view angle offset (i.e.\",\n \"the view angle difference between left and right fibers) obtained from cold images.\",\n \"The estimation parameters are the M and F factors (which are uniquely determined by means of the MFDL and MFDR), the loss factor, the index limit, the macro factor for adjustment of LA, the MFDL and MFDR representing MFD of left and right fibers, as well as LS.\",\n \"The loss factor and the index limit are parameters designed for erbium fiber splicing processes only.\",\n \"The basic procedures how to derive the total loss from primary information of the images and the estimation parameters is schematically illustrated in the FIG.\",\n \"2.Off-Line Tuning Procedures For a sequence of n splices, a splice database for the off-line tuning can be created by use of the upload software modules.\",\n \"Assuming that the measured splice loss for the i'th splice is LMiε{LM1LM2, .\",\n \".\",\n \".\",\n \"LMn}, i=-1, 2, .\",\n \".\",\n \".\",\n \"n, and the estimated splice loss corresponding to the i'th splice is LTiε{LT1, LT2, .\",\n \".\",\n \".\",\n \"LTn} the splice database will thus be: Database={LMi, LTi(Pj, Cik)}, i=1, 2, .\",\n \".\",\n \".\",\n \"n; j=1, 2, .\",\n \".\",\n \".\",\n \"; k=1, 2, .\",\n \".\",\n \".\",\n \"(5) where, Pj and Cik are the estimation parameters and the analyzed results obtained from the estimator, respectively.\",\n \"Note particularly that LMi is the measured loss for splice i and shall not be mixed up with the splice loss LM due to core deformation.\",\n \"Further, the estimation parameters Pj, j=1, 2, .\",\n \".\",\n \".\",\n \", are different for different fusion processes used and for different kind of fibers spliced.\",\n \"After uploading all data into the worksheet, it is important to sort the data according to fusion processes being used.\",\n \"There are two reasons for data sort.\",\n \"(1) Some parameters have been designed not only for optimizing the estimator but also for controlling the fusion processes.\",\n \"These parameters should not be optimized during the off-line tuning if they have been used as control parameters in the fusion processes.\",\n \"This is because any optimization of these parameters might change the fusion process completely.\",\n \"The typical parameter in Ericsson FSU995 splicers is the index limit.\",\n \"(2) The models used for different fusion processes in the estimator might be completely different.\",\n \"For instance, in a normal splice, the contributions from the micro bending loss LD dominate the total splice losses.\",\n \"Therefore, the model calculations are based mainly on the LD.\",\n \"On the other hand, however, if the core offset of two fibers is large, e.g.\",\n \">1 μm, the splice losses will strongly depend not only on LD but also on the wavelength of the signal being transmitted.\",\n \"In this case, a new model for wavelength dependence losses has to be used for fully description of loss mechanisms.\",\n \"The typical example in Ericsson FSU 995 splicers is the estimation processes for the attenuator maker.\",\n \"The Bellcore Criteria for Optimization The basic objective of the off-line tuning is to optimize the estimation parameters so that any, or preferably the best, set of parameters fulfilled the Bellcore splice loss estimator accuracy criteria can be found.\",\n \"The Bellcore accuracy criteria is defined for 90% and 100% of splice populations, respectively, in Table 1: TABLE 1 Bellcore Accuracy Criteria for Splice loss estimator Estimating range Requirement Objective Actual loss 90% 100% 90% 100% ≦0.40 dB ±0.10 dB ±0.25 dB ±0.05 dB ±0.20 dB >0.40 dB ±25% ±50% ±15% ±30% Based on the Bellcore criteria, the algorithms for evaluation of loss estimation can be derived.\",\n \"As an example, the Bellcore criteria used for the off-line tuning in Ericsson FSU995 splicer is given below: Li,min(LMi)≦LTi(P1, P2, P3, P4, P5, P6, Ci1, Ci2, Ci3)≦Li,max(LMi); i=1, 2, .\",\n \".\",\n \".\",\n \", n (6) where P1, P2, P3, P4, P5, P6 are the estimation parameters (i.e.\",\n \"MFDL, MFDR, loss factor, index limit, macro factor and the LS) for a given fusion process and given kind of fiber.\",\n \"Similarly, Ci1, Ci2, Ci3 are the analyzed results (i.e.\",\n \"LD, V, and the view angle offset).\",\n \"For the i'th splice, Cik (k=1,2,3) are given (i.e.\",\n \"derived from hot an cold images, respectively), whereas LTi can be varied by selecting suitable set of Pj (j=1,2 .\",\n \".\",\n \".\",\n \"6).\",\n \"The tunable range of LTi can be controlled by criteria parameters Li,max and Li,min(LMi).\",\n \"For simplicity, we assume, here, that the maximum splice loss is 2.0 dB.\",\n \"Thus, the criteria for the i'th'splice are given by the following relations.\",\n \"For 90% of splice population: Li,maxLMi)=LMi+0.10 if Lmi≦0.4 dB LMi+0.25 LMI if 0.4<LMI≦2.0 dB (7) Li,min(LMi)=LMi−0.10 if LMI≦0.4 dB LMi−0.25 LMi if 0.4<LMi≦2.0 dB (8) For 100% of splice population: Li,max(LMi+0.25 if LMi≦0.4 dB LMi+0.50 LMi if 0.4<LMi≦2.0 dB (9) Li,min(LMi)=LMi−0.25 if LMi≦0.4 dB lMi−0.50 LMi if 0.4<LMi≦2.0 dB (10) In principle, the estimation parameters could be optimized by continuously varying all parameters Pj with well-defined optimization routines and simultaneously calculating LTi with formulas (1)-(4) until the Bellcore criteria, i.e.\",\n \"formula (6), is fulfilled for all splices.\",\n \"FIG.\",\n \"3 shows a typical example of the optimized data that fulfills the Bellcore requirement.\",\n \"The areas within dashed lines and solid lines, respectively, represent the 90% and the 100% Bellcore criteria, respectively.\",\n \"In this particular case, both 90% and 100% Bellcore criteria are completely fulfilled.\",\n \"Regression Analysis Criteria for the Best Solution It can be noticed that a set of optimized parameters that fulfill the Bellcore criteria is not unique.\",\n \"Different set of optimized parameters that fulfill the Bellcore criteria may be quite different, depending on optimization routines and parameter combinations being used for optimization.\",\n \"Though, even if any solution that fulfills the Bellcore criteria should be considered as an acceptable solution, it is still an open question whether it is interesting to select the best solution within Bellcore criteria.\",\n \"The best solution could be defined as the set of parameters that maximize the probability that the estimated slice loss data represent the measured splice loss data, i.e.\",\n \"the data distribution in FIG.\",\n \"3 should be optimized to be as closed to the diagonal line (dash-doted line in FIG.\",\n \"3) as possible.\",\n \"In order to search for the best solution, the method of regression analysis is used.\",\n \"Using such method, a regression line is generated by the “least-square fitting method” to give the best fit for the data set {LMi, LTi(Pj,Cik)}.\",\n \"The regression line can be expressed by: LT=aLM+b (11) where: a = n ( ∑ i = 1 n L Mi L Ti ) - ( ∑ i = 1 n L Mi ) ( ∑ i = 1 n L Ti ) n { ∑ i = 1 n ( L Mi 2 ) } - ( ∑ i = 1 n L Mi ) 2 and ( 12 ) b = ( ∑ i = 1 n L Ti ) { ∑ i = 1 n ( L Mi 2 ) } - ( ∑ i = 1 n L Mi ) ( ∑ i = 1 n L Mi L Ti ) n { ∑ i = 1 n ( L Mi 2 ) } - ( ∑ i = 1 n L Mi ) 2 ( 13 ) Thus, the criteria for the best solution is the data set that not only fulfill the Bellcore criteria but the regression analysis criteria as well.\",\n \"The regression analysis criteria is given by |a−1|≦ε1 |b|≦ε2 (14) where, a and b are the slope and the interception of the regression line.\",\n \"ε1 and ε2 are the infinitesimal constants that set the conditions for controlling and terminating the optimization procedures.\",\n \"In practice, however, additional criteria have to be taken into consideration, e.g.\",\n \"computing time restraints.\",\n \"Since many factors (e.g.\",\n \"hot spots, arc sparks) can not be completely handled by the estimator, it might not be possible to optimize the estimation parameters so that the data set completely fulfill the Bellcore criteria within finite computing time.\",\n \"Therefore, the criteria with well-defined computing loops and/or computing time have to be employed to terminate optimization processes if necessary.\",\n \"In FIGS.\",\n \"4-6, the concept of the above-described two-step optimization is demonstrated.\",\n \"FIG.\",\n \"4 shows the original data set that are uploaded without any optimization.\",\n \"FIG.\",\n \"5 shows that the first step of optimization that fulfills the Bellcore criteria.\",\n \"In the second step of optimization, shown in FIG.\",\n \"6, the best solution is found by use of regression analysis criteria.\",\n \"Program Flowing Chart Based on the basic concepts and techniques discussed above, a simplified program flowing chart used for the off-line tuning in the Ericsson FSU995 splicer is given in FIG.\",\n \"7.A splice database {LMi, LTi(Pj, Cik)} is in a step 71 (which may indeed include a plurality of sub-steps) imported into the offline tuning model 21 of the apparatus 5 as shown in FIG.\",\n \"1.Then, in a step 73, the estimation parameters Pj are altered and W is calculated by using formulas (1)-(4).\",\n \"In a step 75 it is checked whether LTi fulfills formula (6) and if not formula (6) is fulfilled the algorithm is returned to step 73.If formula (6) is fulfilled, then in a step 77 a regression line LT=aLM+b is generated.\",\n \"Then, in a step 79, it is checked whether formula 14 is fulfilled and if not formula (14) is fulfilled the algorithm is returned to step 73.If formula (14) is fulfilled, the estimation parameters are exported, in a step 81, to the download module 17 of FIG.\",\n \"1, whereafter the algorithm is ended.\",\n \"Optionally, in a step 83 a splice loss diagram is created similar to those shown in FIGS.\",\n \"3 and 5-6.It will be obvious that the invention may be varied in a plurality of ways.\",\n \"Such variations are not to be regarded as a departure from the scope of the invention.\",\n \"All such modifications as would be obvious to one skilled in the art are intended to be included within the scope of the appended claims.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473766"}}},{"rowIdx":750,"cells":{"text":{"kind":"list like","value":[["Alphanubeta3 integrin-binding polypeptide monobodies and their use","The present invention relates to a polypeptide monobody which includes a modified acid sequence and renders the polypeptide monobody able to bind selectively to αvβ3 integrin.","Fusion proteins and conjugates which include the polypeptide monobody, as well as compositions containing the polypeptide monobody, fusion proteins, or conjugates are also disclosed.","Uses thereof include: treating or preventing an αvβ3 integrin-mediated disease or disorder, inhibiting αvβ3 integrin activity, treating a cancerous or precancerous condition, imaging tissues using positron emission tomography or magnetic resonance imaging, assessing the metastatic characteristics of a tumor, and delivering DNA to a cell."],["1.A polypeptide monobody comprising: at least two Fn3 β-strand domain sequences with a loop region sequence linked between adjacent β-strand domain sequences; and optionally, an N-terminal tail of at least about 2 amino acids, a C-terminal tail of at least about 2 amino acids, or both; wherein at least one loop region sequence, the N-terminal tail, or the C-terminal tail comprises a modified amino acid sequence which varies by deletion, insertion, or replacement of at least two amino acids from a corresponding loop region, N-terminal tail, or C-terminal tail in a wild-type Fn3 domain of fibronectin, and wherein the polypeptide monobody binds selectively to αvβ3 integrin.","2.The polypeptide monobody according to claim 1, wherein the modified amino acid sequence comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).","3.The polypeptide monobody according to claim 2, wherein the modified amino acid sequence comprises LRGDWSED (SEQ ID No: 5), VRGDWYEY (SEQ ID No: 6), GRGDWTEH (SEQ ID No: 10), ARGDWVEG (SEQ ID No: 11), PRGDWTEG (SEQ ID No: 12), PRGDWIEF (SEQ ID No: 15), PRGDWNEG (SEQ ID No: 23), or FRGDWIEL (SEQ ID No: 24).","4.The polypeptide monobody according to claim 1, wherein said at least one loop region sequence is selected from the group consisting of a BC loop region sequence, a DE loop region sequence, an FG loop region sequence, and combinations thereof.","5.The polypeptide monobody according to claim 4, wherein said at least one loop region sequence is an FG loop region sequence.","6.The polypeptide monobody according to claim 5, wherein the FG loop region sequence comprises, as the modified amino acid sequence, XRGDWXEX where X is any amino acid (SEQ ID No: 26).","7.The polypeptide monobody according to claim 6, wherein the modified amino acid sequence comprises LRGDWSED (SEQ ID No: 5), VRGDWYEY (SEQ ID No: 6), GRGDWTEH (SEQ ID No: 10), ARGDWVEG (SEQ ID No: 11), PRGDWTEG (SEQ ID No: 12), PRGDWIEF (SEQ ID No: 15), PRGDWNEG (SEQ ID No: 23), or FRGDWIEL (SEQ ID No: 24).","8.A nucleic acid molecule encoding the polypeptide monobody of claim 1.9.The nucleic acid molecule according to claim 8, wherein the nucleic acid is DNA.","10.A DNA construct comprising: the DNA molecule of claim 9; a promoter-effective DNA molecule operably coupled 5′ of the DNA molecule; and a transcription termination DNA molecule operably coupled 3′ of the DNA molecule.","11.An expression vector into which is inserted a DNA construct according to claim 9.12.A host cell transformed with a DNA construct according to claim 10.13.The host cell according to claim 12, wherein the host cell is a prokaryote.","14.The host cell according to claim 12, wherein the host cell is a eukaryote.","15.A fusion protein comprising: a polypeptide monobody according to claim 1 and a second polypeptide linked by peptide bond to the polypeptide monobody, the second polypeptide being (i) an epitope tag polypeptide, (ii) a detectable marker polypeptide, (iii) a metal ion-complexing polypeptide, or (iv) a DNA-binding polypeptide.","16.The fusion protein according to claim 15 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX (SEQ ID No: 26) where X is any amino acid.","17.The fusion protein according to claim 15 wherein the second polypeptide is an epitope tag polypeptide.","18.The fusion protein according to claim 17 wherein the epitope tag comprises a polyhistidine amino acid sequence.","19.The fusion protein according to claim 15 wherein the second polypeptide is a detectable marker polypeptide.","20.The fusion protein according to claim 19 wherein the detectable marker polypeptide is an alkaline phosphatase.","21.The fusion protein according to claim 15 wherein the second polypeptide is a metal ion-complexing polypeptide.","22.The fusion protein according to claim 21 wherein the metal-ion complexing polypeptide is a polyhistidine amino acid sequence.","23.The fusion protein according to claim 15 wherein the second polypeptide is a DNA-binding polypeptide.","24.The fusion protein according to claim 23 wherein the DNA-binding polypeptide comprises a polylysine amino acid sequence.","25.A fusion protein-metal ion complex comprising a fusion protein according to claim 21 which is complexed with a metal ion.","26.The fusion protein-metal ion complex according to claim 25 wherein the metal ion is a positron emitting metal ion, a paramagnetic metal ion, a radioactive metal ion, or a non-radioactive metal ion.","27.The fusion protein-metal ion complex according to claim 25 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).","28.A fusion protein-DNA complex comprising a fusion protein according to claim 23 which is complexed with a DNA molecule via the DNA-binding polypeptide.","29.The fusion protein-DNA complex according to claim 28 wherein the DNA-binding polypeptide comprises a polylysine amino acid sequence.","30.The fusion protein-DNA complex according to claim 28 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).","31.A nucleic acid molecule encoding the fusion protein of claim 15.32.The nucleic acid molecule according to claim 31, wherein the nucleic acid is DNA.","33.A DNA construct comprising: the DNA molecule of claim 32; a promoter-effective DNA molecule operably coupled 5′ of the DNA molecule; and a transcription termination DNA molecule operably coupled 3′ of the DNA molecule.","34.An expression vector into which is inserted a DNA construct according to claim 33.35.A host cell transformed with a DNA construct according to claim 33.36.The host cell according to claim 35, wherein the host cell is a prokaryote.","37.The host cell according to claim 35, wherein the host cell is a eukaryote.","38.A conjugate comprising a polypeptide monobody according to claim 1 conjugated to (i) a chemotherapeutic agent, (ii) a contrasting agent, or (iii) an organic chelating agent.","39.The conjugate according to claim 38 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).","40.The conjugate according to claim 38 wherein the polypeptide monobody is conjugated to a chemotherapeutic agent.","41.The conjugate according to claim 38 wherein the polypeptide monobody is conjugated to a contrasting agent or an organic chelating agent.","42.The conjugate according to claim 41, wherein the contrasting agent is a dextran-coated superparamagnetic iron oxide particle, a lipid vesicle with Gd3+ attached to a surface thereof, or poly-lysine with Gd3+ attached to an amino group thereof 43.The conjugate according to claim 41, wherein the organic chelating agent is diethylenetriamine pentaacetic acid.","44.A composition comprising a pharmaceutically acceptable carrier and a polypeptide monobody according to claim 1.45.A composition comprising a pharmaceutically acceptable carrier and the fusion protein according to claim 15.46.A composition comprising a pharmaceutically acceptable carrier and the conjugate according to claim 38.47.A method of treating or preventing an αvβ3 integrin-mediated disease or disorder comprising: administering to a patient in need thereof, an effective amount of a polypeptide or polypeptide monobody according to claim 1 which binds to the αvβ3 integrin to inhibit activity thereof, thereby treating or preventing the αvβ3 integrin-mediated disease or disorder.","48.The method according to claim 47 wherein said administering is carried out under conditions which are effective to contact one or more cells expressing αvβ3 integrin with a sufficient amount of the polypeptide monobody to inhibit the activity of αvβ3 integrin on such one or more cells.","49.The method according to claim 47 wherein the αvβ3 integrin-mediated disease or disorder is tumor metastasis, solid tumor growth, osteoporosis, Paget's disease, humoral hypercalcemia of malignancy, angiogenesis, retinopathy, arthritis, periodontal disease, psoriasis, smooth muscle cell migration, viral infection, fungal infection, and bacterial infection.","50.The method according to claim 47 wherein the αvβ3 integrin-mediated disease or disorder is tumor metastasis or solid tumor growth, the polypeptide monobody being present in the form of a conjugate comprising the polypeptide monobody conjugated to a chemotherapeutic agent.","51.The method according to claim 47 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).","52.A method of treating a cancerous or precancerous condition comprising: administering to a patient in need thereof an effective amount of a polypeptide monobody according to claim 1, wherein the polypeptide monobody binds to cancerous or precancerous cells expressing αvβ3 integrin to inhibit αvβ3 integrin-induced activity of the cancerous or precancerous cells.","53.The method according to claim 52 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).","54.A method of treating a cancerous or precancerous condition comprising: administering to a patient in need thereof an effective amount of a conjugate according to claim 40, wherein the polypeptide monobody binds to cancerous or precancerous cells expressing αvβ3 integrin to inhibit αvβ3 integrin-induced activity of the cancerous or precancerous cells and delivers the chemotherapeutic agent to the cancerous or precancerous cells.","55.The method according to claim 54 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).","56.A method of imaging tissues using positron emission tomography or magnetic resonance imaging, the method comprising: administering a fusion protein-metal ion complex according to claim 25 to a patient and detecting, by positron emission tomography or magnetic resonance imaging, whether the conjugate is localized within any tissues of the patient.","57.The method according to claim 56 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).","58.A method of imaging tissues using positron emission tomography or magnetic resonance imaging, the method comprising: administering a conjugate according to claim 41 to a patient and detecting, by positron emission tomography or magnetic resonance imaging, whether the conjugate is localized within any tissues of the patient.","59.The method according to claim 58 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).","60.A method of assessing the metastatic characteristics of a tumor, the method comprising: administering a fusion protein-metal ion complex according to claim 25 to a patient and detecting, by positron emission tomography or magnetic resonance imaging, whether the conjugate is localized within any tissues of the patient, wherein localization of the fusion protein-metal ion complex indicates an angiogenic site containing a potentially metastatic tumor.","61.The method according to claim 60 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).","62.A method of assessing the metastatic characteristics of a tumor, the method comprising: administering a conjugate according to claim 41 to a patient and detecting, by positron emission tomography or magnetic resonance imaging, whether the conjugate is localized within any tissues of the patient, wherein localization of the conjugate indicates an angiogenic site containing a potentially metastatic tumor.","63.The method according to claim 62 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).","64.A method of inhibiting αvβ3 integrin activity comprising: contacting αvβ3 integrin with a polypeptide monobody according to claim 1 under conditions effective to inhibit the activity of the αvβ3 integrin.","65.The method according to claim 64 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).","66.A method of delivering DNA to a cell comprising: providing a fusion protein-DNA complex according to claim 28 and contacting a cell expressing an αvβ3 integrin with the fusion protein-DNA complex under conditions effective to cause uptake of the fusion protein-DNA complex, thereby delivering the DNA into the cell.","67.The method according to claim 66 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).","68.An isolated polypeptide which binds to an αvβ3 integrin and comprises the amino acid sequence XRGDWXEX where X is any amino acid (SEQ ID No: 26).","69.The polypeptide according to claim 68, wherein the polypeptide comprises the amino acid sequence of LRGDWSED (SEQ ID No: 5), VRGDWYEY (SEQ ID No: 6), GRGDWTEH (SEQ ID No: 10), ARGDWVEG (SEQ ID No: 11), PRGDWTEG (SEQ ID No: 12), PRGDWIEF (SEQ ID No: 15), PRGDWNEG (SEQ ID No: 23), or FRGDWIEL (SEQ ID No: 24)."],["<SOH> BACKGROUND OF THE INVENTION <EOH>αvβ3 (CD51/CD61) is a member of the integrin family of cell surface adhesion receptors.","Over 20 different αβ integrin heterodimers exist, each with different tissue and ligand specificities.","Normal tissue distribution of αvβ3 is generally limited to high levels of expression on osteoclasts, with lower levels observed on platelets, megakaryocytes, kidney, vascular smooth muscle, placenta, dendritic cells (Weiss et al., 2001), and in varying amounts on normal endothelium (reviewed in Horton, 1997).","In contrast to α5β1, which binds to only fibronectin, αvβ3 binds to a wide range of RGD-containing integrin ligands, including but not limited to fibronectin, vitronectin, osteopontin, von Willebrandt factor, and fibrinogen (Horton, 1997).","αvβ3 integrin is a multifunctional cell surface receptor that has pleiotropic roles in normal cell growth and survival.","αvβ3 integrin can also contribute to oncogenesis.","Consistent with this, upregulation of αvβ3 expression has been observed on the endothelial cells of angiogenic vessels, and binding of αvβ3 to the basement membrane is a critical step in the angiogenesis induced by basic fibroblast growth factor and tumor necrosis factor-α (Friedlander et al., 1995).","Expression of αvβ3 has also been implicated in tumor invasion, and it has been shown that αvβ3 binds matrix metalloproteinase-2 (MM-2) and presents MMP-2 on the surface of invasive carcinomas and on invasive angiogenic endothelial cells (Brooks et al., 1996; Silletti et al., 2001).","αvβ3 also regulates cell growth and survival, since ligation of this receptor can, under some circumstances, induce apoptosis in tumor cells (Kozlova et al., 2001).","Furthermore, disruption of cell adhesion with anti-αvβ3 antibodies, RGD peptides, and other integrin antagonists has been shown to slow tumor growth (Chatterjee et al., 2000; Chatterjee et al., 2001; and Brooks et al., 1996).","Finally, the selective upregulation of αvβ3 expression on tumor blood vessels is also being explored as the basis for imaging of neoplastic lesion, and the αvβ3-specific antibody LM609 has been successfully used for this purpose in vivo (Sipkins et al., 1998).","Novel molecules capable of binding with high specificity to αvβ3 integrin have potential utility in several applications, and as a consequence, αvβ3 has been a frequent target for drug discovery and selection of new binding ligands.","Phage display technology, in which combinatorial peptide libraries are expressed on the surface of bacteriophages, has been used to select for peptide ligands capable of binding to αvβ3—yielding a wide range of RGD-containing peptide sequences capable of interacting at moderate affinity with αvβ3 and other integrins (Koivunen et al., 1994; Healy et al., 1995).","Phage-displayed random peptide libraries have also been constructed and screened using framework proteins such as the cytotoxic T lymphocyte-associated antigen 4 (CTLA-4).","This resulted in the identification of phage clones which could be used to stain human umbilical vein endothelial cells in a flow cytometric assay.","However, the ability of the purified recombinant CTLA-4 protein to stain cells, and its cross-reactivity with other integrins was not reported (Hufton et al., 2000).","Fibronectin is a natural ligand of integrins, and it contains repeats of three types of domains.","The tenth fibronectin type III domain (FNfn10) includes the RGD sequence in the loop connecting the F and G β-strands (FG loop) (Main et al., 1992).","FNfn10 was developed as a scaffold for phage display of peptides because of its small size (94 residues), monomeric assembly, and ability to retain its global fold while exposed loops were randomized (Koide et al., 1998).","In addition, FNfn10 lacks cysteine residues and requires no post-translational modification, allowing for large-scale bacterial expression.","It has been shown that residues in the FG loop including the RGD sequence are highly flexible (Main et al., 1992; Carr et al., 1997) and this flexibility of the FG loop has been implicated as the origin of the ability of FNfn10 to interact with multiple integrins (Main et al., 1992).","While the stability of monobodies makes them well suited for intracellular studies, there has been no prior use of monobodies to probe for modified FG loop or other loop sequences with enhanced binding affinity for and selectivity between integrins, particularly αvβ3 integrin.","The present invention overcomes these and other deficiencies in the art."],["<SOH> SUMMARY OF THE INVENTION <EOH>A first aspect of the present invention relates to a polypeptide monobody which includes: at least two Fn3 β-strand domain sequences with a loop region sequence linked between adjacent β-strand domain sequences; and optionally, an N-terminal tail of at least about 2 amino acids, a C-terminal tail of at least about 2 amino acids, or both; wherein at least one loop region sequence, the N-terminal tail, or the C-terminal tail comprises a modified amino acid sequence which varies by deletion, insertion, or replacement of at least two amino acids from a corresponding loop region, N-terminal tail, or C-terminal tail in a wild-type Fn3 domain of fibronectin, and wherein the polypeptide monobody binds selectively to αvβ3 integrin.","Compositions containing the polypeptide monobody are also disclosed.","A second aspect of the present invention relates to a nucleic acid molecule encoding a polypeptide monobody of the present invention.","A third aspect of the present invention relates to a DNA construct which includes a DNA molecule encoding a polypeptide monobody of the present invention; a promoter-effective DNA molecule operably coupled 5′ of the DNA molecule; and a transcription termination DNA molecule operably coupled 3′ of the DNA molecule.","Expression vectors and host cells which include the DNA construct are also disclosed.","A fourth aspect of the present invention relates to a fusion protein which includes: a polypeptide monobody of the present invention and a second polypeptide linked by peptide bond to the polypeptide monobody, the second polypeptide being (i) an epitope tag polypeptide, (ii) a detectable marker polypeptide, (iii) a metal ion-complexing polypeptide, or (iv) a DNA-binding polypeptide.","Compositions containing the fusion protein are also disclosed.","A fifth aspect of the present invention relates to a fusion protein-metal ion complex which includes a fusion protein of the present invention including a metal ion-complexing polypeptide as the second polypeptide, where the metal ion-complexing polypeptide is complexed with a metal ion.","A sixth aspect of the present invention relates to a fusion protein-DNA complex which includes a fusion protein of the present invention including a DNA-binding polypeptide as the second polypeptide, where the DNA-binding polypeptide is complexed with a DNA molecule via the DNA-binding polypeptide.","A seventh aspect of the present invention relates to a nucleic acid molecule encoding a fusion protein of the present invention.","An eighth aspect of the present invention relates to a DNA construct which includes a DNA molecule encoding a fusion protein of the present invention; a promoter-effective DNA molecule operably coupled 5′ of the DNA molecule; and a transcription termination DNA molecule operably coupled 3′ of the DNA molecule.","Expression vectors and host cells which include the DNA construct as also disclosed.","A ninth aspect of the present invention relates to a conjugate comprising a polypeptide monobody of the present invention conjugated to (i) a chemotherapeutic agent, (ii) a contrasting agent, or (iii) an organic chelating agent.","Compositions containing the conjugate are also disclosed.","A tenth aspect of the present invention relates to a method of treating or preventing an αvβ3 integrin-mediated disease or disorder which includes: administering to a patient in need thereof an effective amount of a polypeptide monobody of the present invention which binds to the αvβ3 integrin to inhibit activity thereof, thereby treating or preventing the αvβ3 integrin-mediated disease or disorder.","An eleventh aspect of the present invention relates to a method of treating a cancerous or precancerous condition which includes: administering to a patient in need thereof an effective amount of a polypeptide monobody of the present invention, wherein the polypeptide monobody binds to cancerous or precancerous cells expressing αvβ3 integrin to inhibit αvβ3 integrin-induced activity of the cancerous or precancerous cells.","A twelfth aspect of the present invention relates to a method of treating a cancerous or precancerous condition which includes: administering to a patient in need thereof an effective amount of a conjugate of the present invention, wherein the polypeptide monobody binds to cancerous or precancerous cells expressing αvβ3 integrin to inhibit αvβ3 integrin-induced activity of the cancerous or precancerous cells and delivers the chemotherapeutic agent to the cancerous or precancerous cells.","A thirteenth aspect of the present invention relates to a method of imaging tissues using positron emission tomography or magnetic resonance imaging, the method including: administering a fusion protein-metal ion complex of the present invention to a patient and detecting, by positron emission tomography or magnetic resonance imaging, whether the complex is localized within any tissues of the patient.","A fourteenth aspect of the present invention relates to a method of imaging tissues using positron emission tomography or magnetic resonance imaging, the method including: administering to a patient a conjugate of the present invention which includes a contrasting agent and detecting, by positron emission tomography or magnetic resonance imaging, whether the conjugate is localized within any tissues of the patient.","A fifteenth aspect of the present invention relates to a method of assessing the metastatic characteristics of a tumor, the method including: administering a fusion protein-metal ion complex of the present invention to a patient and detecting, by positron emission tomography or magnetic resonance imaging, whether the conjugate is localized within any tissues of the patient, wherein localization of the fusion protein-metal ion complex indicates an angiogenic site containing a potentially metastatic tumor.","A sixteenth aspect of the present invention relates to a method of assessing the metastatic characteristics of a tumor, the method including: administering to a patient a conjugate of the present invention which includes a contrasting agent and detecting, by positron emission tomography or magnetic resonance imaging, whether the conjugate is localized within any tissues of the patient, wherein localization of the fusion protein-metal ion complex indicates an angiogenic site containing a potentially metastatic tumor.","A seventeenth aspect of the present invention relates to a method of inhibiting αvβ3 integrin activity which includes: contacting αvβ3 integrin with a polypeptide monobody of the present invention under conditions effective to inhibit the activity of the αvβ3 integrin.","An eighteenth aspect of the present invention relates to a method of delivering DNA to a cell which includes: providing a fusion protein-DNA complex of the present invention and contacting a cell expressing an αvβ3 integrin with the fusion protein-DNA complex under conditions effective to cause uptake of the fusion protein-DNA complex, thereby delivering the DNA into the cell.","A nineteenth aspect of the present invention relates to an isolated polypeptide which binds to an αvβ3 integrin and includes the amino acid sequence XRGDWXEX where X is any amino acid (SEQ ID No: 26).","The structure of the polypeptide monobodies of the present invention are advantageous for multiple applications because they (1) lack disulfide bonds, rendering them resistant against reducing agents; (2) are very stable even at high temperature; and (3) can be produced in a bacterial expression system with yields of up to 50-100 mg per liter of culture.","Moreover, because they are derived from the endogenous human protein fibronectin, which is non-immunogenic, it is believed that the polypeptide monobodies will not elicit an immune response.","As a small, single-chain molecule, DNA encoding the polypeptide monobodies can be incorporated into gene delivery vectors (e.g.","viral vectors, liposomes) for cell or tissue-specific gene expression.","As a result of these properties, the polypeptide monobodies of the present invention can be used to treat or prevent a number of αvβ3 integrin-associated diseases or disorders as well as for diagnostic imaging of αvβ3 integrin-expressing tissues."],["This application claims the benefit of U.S.","Provisional Patent Application Ser.","No.","60/281,481, filed Apr.","4, 2001, which is hereby incorporated by reference in its entirety.","This invention was made, at least in part, with funding received from (1) the National Institutes of Health grant numbers R29-GM55042, T32-AI07362, and T32-GM07356; and (2) the U.S. Army grant numbers DAMD17-99-1-9361, DAMD17-01-1-0384, and DAMD17-01-1-0385.The U.S. government may have certain rights in this invention.","FIELD OF THE INVENTION The present invention relates generally to polypeptide monobodies, more particularly polypeptide monobodies derived from the tenth fibronectin type III domain from human fibronectin (“FNfn10”) which have activity in binding to αvβ3 integrin, fusion proteins or conjugates which include such a polypeptide monobody, as well as the uses thereof for modulating αvβ3 integrin-mediated activity.","BACKGROUND OF THE INVENTION αvβ3 (CD51/CD61) is a member of the integrin family of cell surface adhesion receptors.","Over 20 different αβ integrin heterodimers exist, each with different tissue and ligand specificities.","Normal tissue distribution of αvβ3 is generally limited to high levels of expression on osteoclasts, with lower levels observed on platelets, megakaryocytes, kidney, vascular smooth muscle, placenta, dendritic cells (Weiss et al., 2001), and in varying amounts on normal endothelium (reviewed in Horton, 1997).","In contrast to α5β1, which binds to only fibronectin, αvβ3 binds to a wide range of RGD-containing integrin ligands, including but not limited to fibronectin, vitronectin, osteopontin, von Willebrandt factor, and fibrinogen (Horton, 1997).","αvβ3 integrin is a multifunctional cell surface receptor that has pleiotropic roles in normal cell growth and survival.","αvβ3 integrin can also contribute to oncogenesis.","Consistent with this, upregulation of αvβ3 expression has been observed on the endothelial cells of angiogenic vessels, and binding of αvβ3 to the basement membrane is a critical step in the angiogenesis induced by basic fibroblast growth factor and tumor necrosis factor-α (Friedlander et al., 1995).","Expression of αvβ3 has also been implicated in tumor invasion, and it has been shown that αvβ3 binds matrix metalloproteinase-2 (MM-2) and presents MMP-2 on the surface of invasive carcinomas and on invasive angiogenic endothelial cells (Brooks et al., 1996; Silletti et al., 2001).","αvβ3 also regulates cell growth and survival, since ligation of this receptor can, under some circumstances, induce apoptosis in tumor cells (Kozlova et al., 2001).","Furthermore, disruption of cell adhesion with anti-αvβ3 antibodies, RGD peptides, and other integrin antagonists has been shown to slow tumor growth (Chatterjee et al., 2000; Chatterjee et al., 2001; and Brooks et al., 1996).","Finally, the selective upregulation of αvβ3 expression on tumor blood vessels is also being explored as the basis for imaging of neoplastic lesion, and the αvβ3-specific antibody LM609 has been successfully used for this purpose in vivo (Sipkins et al., 1998).","Novel molecules capable of binding with high specificity to αvβ3 integrin have potential utility in several applications, and as a consequence, αvβ3 has been a frequent target for drug discovery and selection of new binding ligands.","Phage display technology, in which combinatorial peptide libraries are expressed on the surface of bacteriophages, has been used to select for peptide ligands capable of binding to αvβ3—yielding a wide range of RGD-containing peptide sequences capable of interacting at moderate affinity with αvβ3 and other integrins (Koivunen et al., 1994; Healy et al., 1995).","Phage-displayed random peptide libraries have also been constructed and screened using framework proteins such as the cytotoxic T lymphocyte-associated antigen 4 (CTLA-4).","This resulted in the identification of phage clones which could be used to stain human umbilical vein endothelial cells in a flow cytometric assay.","However, the ability of the purified recombinant CTLA-4 protein to stain cells, and its cross-reactivity with other integrins was not reported (Hufton et al., 2000).","Fibronectin is a natural ligand of integrins, and it contains repeats of three types of domains.","The tenth fibronectin type III domain (FNfn10) includes the RGD sequence in the loop connecting the F and G β-strands (FG loop) (Main et al., 1992).","FNfn10 was developed as a scaffold for phage display of peptides because of its small size (94 residues), monomeric assembly, and ability to retain its global fold while exposed loops were randomized (Koide et al., 1998).","In addition, FNfn10 lacks cysteine residues and requires no post-translational modification, allowing for large-scale bacterial expression.","It has been shown that residues in the FG loop including the RGD sequence are highly flexible (Main et al., 1992; Carr et al., 1997) and this flexibility of the FG loop has been implicated as the origin of the ability of FNfn10 to interact with multiple integrins (Main et al., 1992).","While the stability of monobodies makes them well suited for intracellular studies, there has been no prior use of monobodies to probe for modified FG loop or other loop sequences with enhanced binding affinity for and selectivity between integrins, particularly αvβ3 integrin.","The present invention overcomes these and other deficiencies in the art.","SUMMARY OF THE INVENTION A first aspect of the present invention relates to a polypeptide monobody which includes: at least two Fn3 β-strand domain sequences with a loop region sequence linked between adjacent β-strand domain sequences; and optionally, an N-terminal tail of at least about 2 amino acids, a C-terminal tail of at least about 2 amino acids, or both; wherein at least one loop region sequence, the N-terminal tail, or the C-terminal tail comprises a modified amino acid sequence which varies by deletion, insertion, or replacement of at least two amino acids from a corresponding loop region, N-terminal tail, or C-terminal tail in a wild-type Fn3 domain of fibronectin, and wherein the polypeptide monobody binds selectively to αvβ3 integrin.","Compositions containing the polypeptide monobody are also disclosed.","A second aspect of the present invention relates to a nucleic acid molecule encoding a polypeptide monobody of the present invention.","A third aspect of the present invention relates to a DNA construct which includes a DNA molecule encoding a polypeptide monobody of the present invention; a promoter-effective DNA molecule operably coupled 5′ of the DNA molecule; and a transcription termination DNA molecule operably coupled 3′ of the DNA molecule.","Expression vectors and host cells which include the DNA construct are also disclosed.","A fourth aspect of the present invention relates to a fusion protein which includes: a polypeptide monobody of the present invention and a second polypeptide linked by peptide bond to the polypeptide monobody, the second polypeptide being (i) an epitope tag polypeptide, (ii) a detectable marker polypeptide, (iii) a metal ion-complexing polypeptide, or (iv) a DNA-binding polypeptide.","Compositions containing the fusion protein are also disclosed.","A fifth aspect of the present invention relates to a fusion protein-metal ion complex which includes a fusion protein of the present invention including a metal ion-complexing polypeptide as the second polypeptide, where the metal ion-complexing polypeptide is complexed with a metal ion.","A sixth aspect of the present invention relates to a fusion protein-DNA complex which includes a fusion protein of the present invention including a DNA-binding polypeptide as the second polypeptide, where the DNA-binding polypeptide is complexed with a DNA molecule via the DNA-binding polypeptide.","A seventh aspect of the present invention relates to a nucleic acid molecule encoding a fusion protein of the present invention.","An eighth aspect of the present invention relates to a DNA construct which includes a DNA molecule encoding a fusion protein of the present invention; a promoter-effective DNA molecule operably coupled 5′ of the DNA molecule; and a transcription termination DNA molecule operably coupled 3′ of the DNA molecule.","Expression vectors and host cells which include the DNA construct as also disclosed.","A ninth aspect of the present invention relates to a conjugate comprising a polypeptide monobody of the present invention conjugated to (i) a chemotherapeutic agent, (ii) a contrasting agent, or (iii) an organic chelating agent.","Compositions containing the conjugate are also disclosed.","A tenth aspect of the present invention relates to a method of treating or preventing an αvβ3 integrin-mediated disease or disorder which includes: administering to a patient in need thereof an effective amount of a polypeptide monobody of the present invention which binds to the αvβ3 integrin to inhibit activity thereof, thereby treating or preventing the αvβ3 integrin-mediated disease or disorder.","An eleventh aspect of the present invention relates to a method of treating a cancerous or precancerous condition which includes: administering to a patient in need thereof an effective amount of a polypeptide monobody of the present invention, wherein the polypeptide monobody binds to cancerous or precancerous cells expressing αvβ3 integrin to inhibit αvβ3 integrin-induced activity of the cancerous or precancerous cells.","A twelfth aspect of the present invention relates to a method of treating a cancerous or precancerous condition which includes: administering to a patient in need thereof an effective amount of a conjugate of the present invention, wherein the polypeptide monobody binds to cancerous or precancerous cells expressing αvβ3 integrin to inhibit αvβ3 integrin-induced activity of the cancerous or precancerous cells and delivers the chemotherapeutic agent to the cancerous or precancerous cells.","A thirteenth aspect of the present invention relates to a method of imaging tissues using positron emission tomography or magnetic resonance imaging, the method including: administering a fusion protein-metal ion complex of the present invention to a patient and detecting, by positron emission tomography or magnetic resonance imaging, whether the complex is localized within any tissues of the patient.","A fourteenth aspect of the present invention relates to a method of imaging tissues using positron emission tomography or magnetic resonance imaging, the method including: administering to a patient a conjugate of the present invention which includes a contrasting agent and detecting, by positron emission tomography or magnetic resonance imaging, whether the conjugate is localized within any tissues of the patient.","A fifteenth aspect of the present invention relates to a method of assessing the metastatic characteristics of a tumor, the method including: administering a fusion protein-metal ion complex of the present invention to a patient and detecting, by positron emission tomography or magnetic resonance imaging, whether the conjugate is localized within any tissues of the patient, wherein localization of the fusion protein-metal ion complex indicates an angiogenic site containing a potentially metastatic tumor.","A sixteenth aspect of the present invention relates to a method of assessing the metastatic characteristics of a tumor, the method including: administering to a patient a conjugate of the present invention which includes a contrasting agent and detecting, by positron emission tomography or magnetic resonance imaging, whether the conjugate is localized within any tissues of the patient, wherein localization of the fusion protein-metal ion complex indicates an angiogenic site containing a potentially metastatic tumor.","A seventeenth aspect of the present invention relates to a method of inhibiting αvβ3 integrin activity which includes: contacting αvβ3 integrin with a polypeptide monobody of the present invention under conditions effective to inhibit the activity of the αvβ3 integrin.","An eighteenth aspect of the present invention relates to a method of delivering DNA to a cell which includes: providing a fusion protein-DNA complex of the present invention and contacting a cell expressing an αvβ3 integrin with the fusion protein-DNA complex under conditions effective to cause uptake of the fusion protein-DNA complex, thereby delivering the DNA into the cell.","A nineteenth aspect of the present invention relates to an isolated polypeptide which binds to an αvβ3 integrin and includes the amino acid sequence XRGDWXEX where X is any amino acid (SEQ ID No: 26).","The structure of the polypeptide monobodies of the present invention are advantageous for multiple applications because they (1) lack disulfide bonds, rendering them resistant against reducing agents; (2) are very stable even at high temperature; and (3) can be produced in a bacterial expression system with yields of up to 50-100 mg per liter of culture.","Moreover, because they are derived from the endogenous human protein fibronectin, which is non-immunogenic, it is believed that the polypeptide monobodies will not elicit an immune response.","As a small, single-chain molecule, DNA encoding the polypeptide monobodies can be incorporated into gene delivery vectors (e.g.","viral vectors, liposomes) for cell or tissue-specific gene expression.","As a result of these properties, the polypeptide monobodies of the present invention can be used to treat or prevent a number of αvβ3 integrin-associated diseases or disorders as well as for diagnostic imaging of αvβ3 integrin-expressing tissues.","BRIEF DESCRIPTION OF THE DRAWINGS FIG.","1 is a schematic drawing of FNfn10 (Main et al., 1992) showing the RGD sequence and the positions of residues in the FG loop that were diversified in the JCFNRGD library.","This figure was made with the program MOLSCRIPT (Kraulis, 1991).","FIG.","2 illustrates a nucleotide sequence (SEQ ID No: 1) encoding the amino acid sequence (SEQ ID No: 2) of the wild-type FNfn10.The amino acid numbering is according to Main et al.","(1992).","The BC loop region and the FG loop region are shown in boxes.","FIGS.","3A-B illustrate the amino acid sequence of the wild-type FNfn10 (SEQ ID No: 2, FIG.","3A) as well as a mutant FNfn10 (SEQ ID No: 3, FIG.","3B) which has the Asp-7 residue replaced with a non-negatively charged amino acid residue (X), which is preferably either Asn or Lys.","As reported in Koide et al.","(2001), both of these mutations have the effect of promoting greater stability of the mutant FNfn10 at neutral pH as compared to the wild-type FNfn10.FIGS.","4A-B illustrate the FG loop sequences used during phage display screening of a JCFN-RGD library.","M13 Phage displaying a library of FNfn10 clones with partially randomized FG loops (XRGDXXXX, SEQ ID No: 4) were amplified using 0.0, 0.1, or 1 mM IPTG to vary FNfn10 copy number on the phage.","Amplified phage populations were subjected to three rounds of biopanning against wells coated with αvβ3 integrin (5 μg/ml).","FIG.","4A shows the FG loop sequences for five clones, each of which was sequenced from the second round eluate of the library amplified with 1 mM IPTG (2JCAV1-2JCAV5, SEQ ID Nos: 5-9), from the third round eluate of the library amplified with 1 mM IPTG (3JCAV1-5, SEQ ID Nos: 10-14), from the third round eluate of the library amplified with low IPTG (3JCLI1-5, SEQ ID Nos: 15-19), and from the third round eluate of the library amplified with no IPTG (3JCNI1-5, SEQ ID Nos: 20-24).","FIG.","4B illustrates the phage binding assay of the phage clones containing the XRGDWXEX consensus (SEQ ID No: 26) and unmodified FNfn10 (pVIII-FNfn10), all amplified without IPTG.","Phage inputs of 109, 108, or 107 pfu were added to wells coated with 5 μg/ml αvβ3 (solid bars) or 0.5 mg/ml BSA (patterned bars).","After one hour, nonbinding phage were removed by extensive rinsing and bound phage were eluted with 0.2M glycine, pH 2.2.Eluates from each well were titered.","Results shown are representative of three different experiments that yielded similar results.","FIGS.","5A-D are graphs illustrating the assessment of FNfn10-3JCLI4 binding affinity to αvβ3 integrin by ELISA.","To determine the half-maximal binding concentration of biotinylated FNfn10-3JCLI4 versus biotinylated FNfn10-WT, wells of a 96-well plate were coated αvβ3 (5 μg/ml), α1β1 (5 μg/ml), or BSA (0.5 mg/ml) and blocked with BSA (FIG.","5A).","Biotinylated FNfn10-3JCLI4 and FNfn10-WT were then added to the wells and allowed to bind for two hours.","Bound protein was detected by streptavidin-horseradish peroxidase (2 μg/ml) and ABTS substrate.","O.D.400 measurements were recorded for biotinylated FNfn10-3JCLI4 binding to αvβ3 (⋄), to α1β1 (♦), or to BSA (×) and biotinylated FNfn10-WT binding to αvβ3 (□).","Error bars reflect the standard deviation between triplicate wells.","To examine protein dissociation over a prolonged (24 hour) time period, a 96-well plate was coated with 5 μg/ml αvβ3 integrin and then incubated with 0.02 μg/ml biotinylated FNfn10-3JCLI4 for two hours; as a control, 10 mM EDTA was added to certain wells, in order to calculate the background level of FNfn10-3JCLI4 binding (FIG.","5B).","After rinsing, 2 μg/ml unlabelled FNfn10-3JCLI4 was added to the preformed protein complexes present in the wells 24, 14, 4, 2 and 1 hours prior to detection of remaining plate-bound biotinylated FNfn10-3JCLI4 using streptavidin-horseradish peroxidase and ABTS.","Baseline binding activity (100%) was calculated as the amount of biotinylated FNfn10-3JCLI4 which remained bound to the plate following incubation of the well for 24 hours, in the presence of buffer alone (i.e., in the absence of any competitor protein), and the O.D.400 of FNfn10-3JCLI4 bound in the presence of 10 mM EDTA was subtracted as background.","All incubations with competitor (background-subtracted) are expressed as a percentage of the O.D.400 of baseline binding (background-subtracted).","Error bars reflect the standard deviation between triplicate wells.","To examine rate of association of αvβ3 and biotinylated FNfn10-3JCLI4, a 96-well plate coated with 5 μg/ml αvβ3 integrin was blocked with BSA (FIG.","5C).","0.01 (▴), 0.02 (▪), or 0.05 (♦) μg/ml of biotinylated FNfn10-3JCLI4 was then added to the coated wells, at 240, 30, 10, 3, and 1 minute prior to washing and detection.","After detection with streptavidin-horseradish peroxidase/ABTS, the O.D.400 of the plate was recorded.","Error bars reflect the standard deviation between triplicate wells.","A 96-well plate was coated with αvβ3, αvβ5, αIIbβ3, α1β1, and α5β1 purified integrins, all at 5 μg/ml (FIG.","5D).","Biotinylated FNfn10-3JCLI4 (dark bars) or FNfn10-WT (light bars) was then added at a concentration of 0.01 μg/ml, and allowed to incubate for 2 hours, followed by washing and detection.","Error bars reflect the standard deviation between triplicate wells.","In FIGS.","5A-D, the experiments shown are all representative of three different experiments that yielded similar results.","FIGS.","6A-D are graphs illustrating competition ELISAs to determine relative affinities of FNfn10-3JCLI4, echistatin, FNfn10-WT, and GRGDSPK peptide (SEQ ID No: 27).","A BSA-blocked 96-well plate which was coated with αvβ3 at 5 μg/ml, after which biotinylated FNfn10-3JCLI4 was then added in the presence or absence of a 0.01, 0.1, 1, 10, or 100-fold molar excess of unlabelled competitor protein, and incubated for two hours.","After washing and detection, the O.D.400 of the plate was recorded.","Error bars reflect the standard deviation between triplicate wells, and the results shown are representative of three different experiments that yielded similar results.","A background subtraction was performed on all data points, by subtracting the measured O.D.400 value that corresponded to binding of biotinylated FNfn10-3JCLI4 in the presence of 10 mM EDTA.","All values in the figure are expressed as a percentage of the O.D.400 of biotinylated FNfn10-3JCLI4 binding in the absence of competitor.","FIG.","6A illustrates the binding competition with unlabelled FNfn10-3JCLI4 (♦) or unlabelled FNfn10-WT (▪).","FIG.","6B illustrates the binding competition with unlabelled FNfn10-3JCLI4 (▴), echistatin (▪), and GRGDSPK peptide (♦)(SEQ ID No: 27).","FIGS.","7A-C are graphs illustrating fluorescence activated cell sorting (FACS) analysis of FNfn10-3JCLI4 binding to human cells.","Biotinylated FNfn10-3JCLI4 or FNfn10-WT were added to 105 cells at a concentration of 1 μg/ml and incubated for 30 minutes.","Bound proteins were then detected with streptavidin-APC, and the mean fluorescence of ten thousand cells per sample was recorded.","All experiments were performed three times, with similar results; error bars (FIGS.","7B-C) represent the standard deviation of three samples of 105 cells stained separately.","FIG.","7A is a histogram plot of flow cytometric staining analysis of K562 and K562-αvβ3 cells using FNfn10-3JCLI4.FIG.","7B shows mean APC fluorescence for integrin-transfected K562 cells stained with biotinylated FNfn10-3JCLI4 (dark bars) or FNfn10-WT (light bars).","To determine the effect of high temperature on FNfn10-3JCLI4 stability, 1 μg/ml 3JCLI4 (♦) or 5 μg/ml LM609 (▪) were incubated for 24 hours in FACS buffer at 4, 20, 37, 50, 65, or 75° C. (FIG.","7C).","105 K562-αvβ3 cells were pelleted for each sample and re-suspended in the appropriately treated FNfn10-3JCLI4 or LM609, followed by detection as above.","FIGS.","8A-C illustrate the inhibition of integrin-dependent cell adhesion.","Vitronectin (FIGS.","8A-B) or fibronectin (FIG.","8C), as well as BSA-coated control wells, were incubated with 105 K562 cells, K562-αvβ3 cells, or K562-αIIbβ3 cells for two hours in the presence or absence of the indicated inhibitors.","Non-adherent cells were removed by washing, while adherent cells were stained with crystal violet, solubilized, and quantitated by measuring the O.D.570.The background level of crystal violet in control wells was calculated on the basis of measured O.D.570 for BSA-coated wells (no integrin).","Error bars represent the standard deviation of triplicate wells, and all experiments were performed three times, with similar results.","In FIG.","8A, adhesion of binding of K562-αvβ3 cells to vitronectin is shown.","Inhibitors added include FNfn10-3JCLI4, FNfn10-WT, and LM609 antibody, at the stated final concentrations.","The lane marked “no αvβ3” corresponds to control K562 cells (no αvβ3), while “no VN” denotes cell adherence (K562-αvβ3 cells) measured on control wells (no vitronectin); the lane marked “NT” corresponds to K562-αvβ3 cells which were bound to vitronectin in the absence of any competitor.","In FIG.","8B, adhesion of K562-αvβ3 cells to vitronectin is shown.","Echistatin (▪) and FNfn10-3JCLI4 (♦) were added at final concentrations of 400, 40, and 4 nM, prior to addition of the cells and initiation of the binding assay.","Binding is expressed as a percentage of the O.D.570 recorded for K562-αvβ3 adhesion in the absence of competitor, and a background subtraction was performed for all data points, which corresponded to the measured O.D.570 for BSA-coated wells (no integrin).","In FIG.","8C, adhesion of K562-αIIbβ3 cells to fibronectin is shown.","Echistatin (▪) and FNfn10-3JCLI4 (♦) were added at final concentrations of 400, 40, and 4 nM prior to addition of the cells and initiation of the binding assay.","Binding is expressed as a percentage of the O.D.570 recorded for K562-αIIbβ3 adhesion in the absence of competitor, and a background subtraction was performed for all data points, which corresponded to the measured O.D.570 for BSA-coated wells (no integrin).","DETAILED DESCRIPTION OF THE INVENTION As used herein, “polypeptide monobody” is intended to mean a polypeptide which includes a β-strand domain lacking in disulfide bonds and containing a plurality of β-strands, two or more loop regions each connecting one β-strand to another β-strand, and optionally an N-terminal tail, a C-terminal tail, or both, wherein at least one of the two or more loop regions, the N-terminal tail, or the C-terminal tail is characterized by activity in binding a target protein or molecule.","More specifically, such polypeptide monobodies of the present invention can include three or more loop regions or, even more specifically, four or more loop regions.","The size of such polypeptide monobodies is preferably less than about 30 kDa, more preferably less than about 20 kDa.","The polypeptide monobodies of the present invention are also characterized by specificity for binding to αvβ3 integrin (i.e., enhanced binding specificity for αvβ3 integrin over other integrins, as compared to wild type polypeptide monobodies or wild type fibronectin).","To achieve the specificity in their binding to αvβ3 integrin, the amino acid sequence of the polypeptide monobody has been modified relative to the scaffold used for its construction.","Scaffolds for formation of a polypeptide monobody should be highly soluble and stable.","It is small enough for structural analysis, yet large enough to accommodate multiple binding domains so as to achieve tight binding and/or high specificity for its target.","An exemplary scaffold for formation of a polypeptide monobody is the fibronectin type III domain (Fn3).","Fibronectin is a large protein which plays essential roles in the formation of extracellular matrix and cell-cell interactions; it consists of many repeats of three types (types I, II, and III) of small domains (Baron et al., 1991).","Fn3 itself is the paradigm of a large subfamily (Fn3 family or s-type Ig family) of the immunoglobulin superfamily.","The Fn3 family includes cell adhesion molecules, cell surface hormone and cytokine receptors, chaperoning, and carbohydrate-binding domains (for reviews, see Bork & Doolittle, 1992; Jones, 1993; Bork et al., 1994; Campbell & Spitzfaden, 1994; Harpez & Chothia, 1994).","Crystallographic studies have revealed that the structure of the DNA binding domains of the transcription factor NF-kB is also closely related to the Fn3 fold (Ghosh et al., 1995; Müller et al., 1995).","These proteins are all involved in specific molecular recognition, and in most cases ligand-binding sites are formed by surface loops, suggesting that the Fn3 scaffold is an excellent framework for building specific binding proteins.","The 3D structure of Fn3 has been determined by NMR (Main et al., 1992) and by X-ray crystallography (Leahy et al., 1992; Dickinson et al., 1994).","The structure is best described as a β-sandwich similar to that of antibody VH domain except that Fn3 has seven β-strands (see FIG.","1) instead of nine.","There are three loops on each end of Fn3; the positions of the BC, DE, and FG loops approximately correspond to those of CDR 1, 2 and 3 of the VH domain.","Fn3 is small (˜94 residues), monomeric, soluble, and stable.","It is one of few members of IgSF that do not have disulfide bonds and, therefore, is stable under reducing conditions.","Fn3 has been expressed in E. Coli (Aukhil et al., 1993).","In addition, 17 Fn3 domains are present just in human fibronectin, providing important information on conserved residues which are often important for the stability and folding (see Main et al., 1992; Dickinson et al., 1994).","From sequence analysis, large variations are seen in the BC and FG loops, suggesting that the loops are not crucial to stability.","NMR studies have revealed that the FG loop is highly flexible; the flexibility has been implicated for the specific binding of the 10th Fn3 to α5β1 integrin through the Arg-Gly-Asp (RGD) motif In the crystal structure of human growth hormone-receptor complex (de Vos et al., 1992), the second Fn3 domain of the receptor interacts with growth hormone via the FG and BC loops, suggesting it is feasible to build a binding site using the two loops.","The tenth type III module of fibronectin has a fold similar to that of immunoglobulin domains, with seven β strands forming two antiparallel β sheets, which pack against each other (FIG.","1; Main et al., 1992).","The structure of the type H module includes seven β strands, which form a sandwich of two antiparallel sheets, one containing three strands (ABE) and the other four strands (C′CFG) (Williams & Barclay, 1988).","The β sheet contains residues Glu-9-Thr-14 (A), Ser-17-Asp-23 (B), and Thr-56-Ser-60 (E).","The majority of the conserved residues contribute to the hydrophobic core, with the invariant hydrophobic residues Trp-22 and Try-68 lying toward the N-terminal and C-terminal ends of the core, respectively.","The β strands are much less flexible and appear to provide a rigid framework upon which functional, flexible loops can be built.","The topology is similar to that of immunoglobulin C domains.","Preferred polypeptide monobodies of the present invention are fibronectin type III (Fn3)-derived polypeptide monobodies.","Fn3 monobodies include at least two Fn3 β-strand domain sequences with a loop region sequence linked between adjacent β-strand domain sequences and optionally, an N-terminal tail of at least about 2 amino acids, a C-terminal tail of at least about 2 amino acids, or both.","The loop region sequence, the N-terminal tail, or the C-terminal tail, or combinations thereof include an amino acid sequence which has binding specificity for αvβ3 integrin.","To render a loop region sequence, N-terminal tail, or C-terminal tail capable of binding to αvβ3 integrin, either the loop region sequence, the N-terminal tail, the C-terminal tail, or a combination thereof varies by deletion, insertion, or replacement of at least two amino acids from a corresponding loop region, N-terminal tail, or C-terminal tail in a wild-type or mutant Fn3 scaffold.","One preferred wild-type Fn3 scaffold is the tenth Fn3 domain of human fibronectin (FNfn10), which has an amino acid sequence according to SEQ ID No: 2 (FIG.","3A).","One preferred mutant Fn3 scaffold is the tenth Fn3 domain of human fibronectin which has a modified Asp7, which is replaced by a non-negatively charged amino acid residue (i.e., Asn, Lys, etc.)","as shown in FIG.","3B (SEQ ID No: 3).","As reported in Koide et al.","(2001), both of these mutations have the effect of promoting greater stability of the mutant FNfn10 at neutral pH as compared to the wild-type FNfn10.Both the mutant and wild-type FNfn10 are characterized by the same structure, namely seven β-strand domain sequences (designated A through G) and six loop regions (AB loop, BC loop, CD loop, DE loop, EF loop, and FG loop) which connect the seven β-strand domain sequences.","In SEQ ID Nos: 2 and 3, the AB loop corresponds to residues 15-16, the BC loop corresponds to residues 22-30, the CD loop corresponds to residues 39-45, the DE loop corresponds to residues 51-55, the EF loop corresponds to residues 60-66, and the FG loop corresponds to residues 76-87.As shown in FIG.","1, the BC loop, DE loop, and FG loop are all located at the same end of the polypeptide monobody.","According to one particular embodiment, the preferred polypeptide monobodies are mutant or wild-type FNfn10 scaffolds which include a modified FG loop region, which polypeptide monobodies exhibit increased activity in binding to αvβ3 integrin.","Such binding is preferably selective of αvβ3 integrin over other integrins.","The modified FG loop region preferably includes an amino acid sequence including XRGDWXEX (SEQ ID No: 26), where X is any amino acid residue.","The polypeptide monobodies of the present invention can be prepared by recombinant techniques, thereby affording the deletion, insertion, or replacement of at least two amino acids from a corresponding loop region, N-terminal tail, or C-terminal tail in a wild-type or mutant Fn3 scaffold.","Deletions can be a deletion of at least two amino acid residues up to substantially all but one amino acid residue appearing in a particular loop region or tail.","Insertions can be an insertion of at least two amino acid residues up to about 25 amino acid residues, preferably at least two up to about 15 amino acid residues.","Replacements can be replacements of at least two up to substantially all amino acid residues appearing in a particular loop region or tail.","The polypeptide monobodies preferably possess an amino acid sequence which is at least 50% homologous to a β-strand domain of the FNfn10.The deletions, insertions, and replacements (relative to wild-type or previously known mutant) on Fn3 scaffolds can be achieved using recombinant techniques beginning with a known nucleotide sequence.","A synthetic gene for the tenth Fn3 of human fibronectin (FIG.","2) was designed which includes convenient restriction sites for ease of mutagenesis and uses specific codons for high-level protein expression (Gribskov et al., 1984).","This gene is substantially identical to the gene disclosed in co-pending U.S. patent application Ser.","No.","09/096,749 to Koide filed Jun.","12, 1998, which is hereby incorporated by reference in its entirety.","The gene was assembled as follows: first the gene sequence was divided into five parts with boundaries at designed restriction sites (FIG.","2); for each part, a pair of oligonucleotides that code opposite strands and have complementary overlaps of about 15 bases was synthesized; the two oligonucleotides were annealed and single strand regions were filled in using the Klenow fragment of DNA polymerase; the double-stranded oligonucleotide was cloned into the pET3a vector (Novagen) using restriction enzyme sites at the termini of the fragment and its sequence was confirmed by an Applied Biosystems DNA sequencer using the dideoxy termination protocol provided by the manufacturer; and these steps were repeated for each of the five parts to obtain the whole gene.","Although this approach takes more time to assemble a gene than the one-step polymerase chain reaction (PCR) method (Sandhu et al., 1992), no mutations occurred in the gene.","Mutations would likely have been introduced by the low fidelity replication by Taq polymerase and would have required time-consuming gene-editing.","Recombinant DNA manipulations were performed according to Molecular Cloning (Sambrook et al., 1989), unless otherwise stated.","To avoid the introduction of mutations during one-step PCR, high fidelity/low error polymerases can be employed as is known in the art.","Desired mutations can be introduced to the Fn3 gene using either cassette mutagenesis, oligonucleotide site-directed mutagenesis techniques (Deng & Nickoloff, 1992), or Kunkel mutagenesis (Kunkel et al., 1987).","Both cassette mutagenesis and site-directed mutagenesis can be used to prepare specifically desired nucleotide coding sequences.","Cassette mutagenesis can be performed using the same protocol for gene construction described above and the double-stranded DNA fragment coding a new sequence can be cloned into a suitable expression vector.","Many mutations can be made by combining a newly synthesized strand (coding mutations) and an oligonucleotide used for the gene synthesis.","Regardless of the approach utilized to introduce mutations into the monobody nucleotide sequence, sequencing can be performed to confirm that the designed mutations (and no other mutations) were introduced by mutagenesis reactions.","In contrast, Kunkel mutagenesis can be utilized to randomly produce a plurality of mutated monobody coding sequences which can be used to prepare a combinatorial library of polypeptide monobodies for screening.","Basically, targeted loop regions (or C-terminal or N-terminal tail regions) can be randomized using the NNK codon (N denoting a mixture of A, T, G, C, and K denoting a mixture of G and T) (Kunkel et al., 1987).","An exemplary DNA molecule encoding a mutant Fn3 polypeptide monobody is set forth as SEQ ID No: 28 below: atgcaggttt ctgatgttcc gcgtgacctg gaagttgttg ctgcgacccc gactagcctg 60 ctgatcagct gggatgctcc tgcagttacc gtgcgttatt accgtatcac gtacggtgaa 120 accggtggta actccccggt tcaggaattc actgtacctg gttccaagtc tactgctacc 180 atcagcggcc tgaaaccggg tgtcgactat accatcactg tatacgctgt tactnnkcgc 240 ggcgatnnkn nknnknnktc caagccaatc tcgattaact accgtaccgg tggcaggttct 300 ggtggcggt 309 In the above DNA sequence, codons nt 235-237, nt 247-249, nt 250-252, nt 253-255, and nt 256-258 (bold typeface) encode random amino acid residues within the FG loop.","According to the preferred embodiments of the present invention, however, codons nt 247-249 and nt 253-255 encode, respectively tryptophan and glutamic acid.","The seven codons extending from nt 230-250 encode a seven residue linker which, though not required for formation of a polypeptide monobody of the present invention, is useful in forming fusion proteins as described below.","Regardless of the approach used to prepare the nucleic acid molecules encoding the polypeptide monobody, the nucleic acid can be incorporated into host cells using conventional recombinant DNA technology.","Generally, this involves inserting the DNA molecule into an expression system to which the DNA molecule is heterologous (i.e., not normally present).","The heterologous DNA molecule is inserted into the expression system or vector in sense orientation and correct reading frame.","The vector contains the necessary elements (promoters, suppressers, operators, transcription termination sequences, etc.)","for the transcription and translation of the inserted protein-coding sequences.","Alternatively, a recombinant gene or DNA construct can be prepared prior to its insertion into an expression vector.","For example, using conventional recombinant DNA techniques, a promoter-effective DNA molecule can be operably coupled 5′ of a DNA molecule encoding the polypeptide monobody and a transcription termination (i.e., polyadenylation sequence) can be operably coupled 3′ thereof.","U.S. Pat.","No.","4,237,224 to Cohen and Boyer describes the production of expression systems in the form of recombinant plasmids using restriction enzyme cleavage and ligation with DNA ligase.","These recombinant plasmids are then introduced by means of transformation and replicated in unicellular cultures including prokaryotic organisms and eukaryotic cells grown in tissue culture.","Recombinant molecules can be introduced into cells via transformation, particularly transduction, conjugation, mobilization, or electroporation.","The DNA sequences are cloned into the vector using standard cloning procedures in the art, as described by Sambrook et al.","(1989).","A variety of host-vector systems may be utilized to express the polypeptide monobody or fusion protein which includes a polypeptide monobody.","Primarily, the vector system must be compatible with the host cell used.","Host-vector systems include but are not limited to the following: bacteria transformed with bacteriophage DNA, plasmid DNA, or cosmid DNA; microorganisms such as yeast containing yeast vectors; and mammalian cell systems infected with virus (e.g., vaccinia virus, adenovirus, etc.).","The expression elements of these vectors vary in their strength and specificities.","Depending upon the host-vector system utilized, any one of a number of suitable transcription and translation elements can be used.","Different genetic signals and processing events control many levels of gene expression (e.g., DNA transcription and messenger RNA (mRNA) translation).","Transcription of DNA is dependent upon the presence of a promoter which is a DNA sequence that directs the binding of RNA polymerase and thereby promotes mRNA synthesis.","The DNA sequences of eukaryotic promoters differ from those of prokaryotic promoters.","Furthermore, eukaryotic promoters and accompanying genetic signals may not be recognized in or may not function in a prokaryotic system and, further, prokaryotic promoters may not be recognized in or may not function in eukaryotic cells.","Promoters suitable for use in each of these systems are well known in the art.","Similarly, translation of mRNA in prokaryotes depends upon the presence of the proper prokaryotic signals which differ from those of eukaryotes.","Efficient translation of mRNA in prokaryotes requires a ribosome binding site called the Shine-Dalgarno (“SD”) sequence on the mRNA.","This sequence is a short nucleotide sequence of mRNA that is located before the start codon, usually AUG, which encodes the amino-terminal methionine of the protein.","The SD sequences are complementary to the 3′-end of the 16S rRNA (ribosomal RNA) and probably promote binding of mRNA to ribosomes by duplexing with the rRNA to allow correct positioning of the ribosome.","For a review on maximizing gene expression, see Roberts & Lauer (1979).","In addition, polyadenylation signals suitable for use in a desired host cell can also be employed to effect appropriate translation of the DNA molecule (encoding the polypeptide monobody of the present invention.","Polyadenylation signals suitable for use in each of the above-identified systems are well known in the art.","Basically, the DNA molecule encoding the polypeptide monobody is ligated to the appropriate promoter and polyadenylation sequences using conventional recombinant techniques, with the promoter being located upstream or 5′ to the DNA molecule and the polyadenylation signal being located downstream or 3′ of the DNA molecule.","As a result of ligating the DNA molecule to the promoter and polyadenylation signal sequences, a DNA construct of the present invention is formed.","This DNA construct can be cloned directly into a suitable expression vector (e.g., plasmid, viral DNA, etc.)","or it can be formed upon cloning the DNA molecule into an empty expression vector which includes an appropriate restriction site intermediate the promoter and polyadenylation signals which are present in the empty vector.","Once the DNA molecule encoding the polypeptide monobody has been cloned into an expression system, it is ready to be incorporated into a host cell.","Such incorporation can be carried out by the various forms of transformation noted above, depending upon the vector/host cell system.","Suitable host cells include, but are not limited to, bacteria, yeast cells, mammalian cells, etc.","Upon growing the host cells in a suitable growth medium, the host cells are encouraged to express the polypeptide monobodies of the present invention.","In at least several of the various uses of the polypeptide monobodies of the present invention, discussed infra, it is often desirable for the polypeptide monobodies to be produced in substantially purified form, particularly when their administration to a patient is contemplated.","Purification can be carried out according to previously reported procedures, which involve metal affinity chromatography for monobodies containing a poly-histidine tag (see Koide et al., 1998).","Another aspect of the present invention relates to a polypeptide (i.e., whether in the form of a polypeptide monobody or not) which includes the amino acid sequence RGDWXE (amino acid residues 2-7 of SEQ ID No: 26) and selectively binds αvβ3 integrin over other integrins.","Such a polypeptide can be prepared, isolated, and purified according to known procedures.","Typically, this involves recombinant expression of the desired polypeptide monobody by a host cell, propagation of the host cells, lysing the host cells, and recovery of supernatant by centrifugation to remove host cell debris.","The supernatant can be subjected to sequential ammonium sulfate precipitation.","The fraction containing the polypeptide monobodies of the present invention is then subjected to gel filtration in an appropriately sized dextran or polyacrylamide column to separate the polypeptide monobodies.","If necessary, the protein fraction may be further purified by HPLC.","A further aspect of the present invention relates to fusion proteins which include a polypeptide or polypeptide monobody of the present invention and a second polypeptide linked by peptide bond to the polypeptide or polypeptide monobody.","The second polypeptide can be (i) an epitope tag polypeptide, e.g., polyhistidine, (ii) a detectable marker polypeptide, e.g., alkaline phosphatase, (iii) a metal ion-complexing polypeptide, or (iv) a DNA-binding polypeptide, e.g., polylysine.","Each of these fusion proteins can be prepared using recombinant techniques described above to form an in-frame gene fusion that can be expressed in appropriate host cells.","Thereafter, the fusion protein can isolated and purified using the techniques described above.","Once the polypeptides, polypeptide monobodies, or fusion proteins have been obtained in isolated or, more preferably, purified form, they can be used for therapeutic, preventative, or diagnostic purposes as described hereinafter.","Because the polypeptides and polypeptide monobodies of the present invention are capable of binding αvβ3 integrin selectively, they can be used for therapeutic, preventative, or diagnostic purposes, or to modify αvβ3 integrin activity.","Although references below are made to use of the polypeptide or polypeptide monobody, it should be appreciated that the polypeptide or polypeptide monobody can also be in the form of a fusion protein or conjugate as described elsewhere herein.","Thus, one aspect of the present invention relates to a method of treating or preventing an αvβ3 integrin-mediated disease or disorder which includes administering to a patient in need thereof, an effective amount of a polypeptide or polypeptide monobody of the present invention which selectively binds to αvβ3 integrin, thereby treating or preventing the αvβ3 integrin-mediated disease or disorder.","Such administration is carried out under conditions which are effective to treat or prevent the αvβ3 integrin-mediated disease or disorder.","Typically, this includes administering the polypeptide or polypeptide monobody to the patient in a manner effective to contact one or more cells expressing αvβ3 integrin with a sufficient amount of the polypeptide or polypeptide monobody to inhibit the activity of αvβ3 integrin on such one or more cells.","As a result of such inhibition, the αvβ3 integrin-mediated diseases or disorders are treated or prevented.","For therapeutic or preventative treatments, the patient can be any mammal which expresses αvβ3 integrin, preferably though not exclusively humans.","Integrins are a group of cell surface glycoproteins that mediate cell adhesion and therefore are mediators of cell adhesion interactions that occur in various biological processes.","Integrins are heterodimers composed of noncovalently linked α and β polypeptide subunits.","Currently at least eleven different α subunits have been identified and at least six different β subunits have been identified.","The various α subunits can combine with various β subunits to form distinct integrins.","The integrin identified as αvβ3 (also known as the vitronectin receptor) has been identified as an integrin that plays a role in various conditions or disease states including tumor metastasis, solid tumor growth (neoplasia), osteoporosis, Paget's disease, humoral hypercalcemia of malignancy, angiogenesis, including tumor angiogenesis, retinopathy, arthritis, including rheumatoid arthritis, periodontal disease, psoriasis and smooth muscle cell migration (e.g.","restenosis).","Additionally, it has been found that integrin inhibiting agents would be useful as antivirals, antifungals and antimicrobials.","Thus, polypeptides or polypeptide monobodies that selectively bind αvβ3 integrin, thereby inhibiting αvβ3 integrin-mediated conditions, would be beneficial for treating or preventing such conditions.","It has been shown that the αvβ3 integrin binds to a number of Arg-Gly-Asp (RGD) containing matrix macromolecules, such as fibrinogen (Bennett et al., 1983), fibronectin (Ginsberg et al., 1983), and von Willebrand factor (Ruggeri et al., 1982).","Compounds containing the RGD sequence mimic extracellular matrix ligands so as to bind to cell surface receptors.","However, it is also known that RGD peptides in general are non-selective for RGD dependent integrins.","For example, most RGD peptides that bind to αvβ3 also bind to αvβ5, αvβ1, and αIIbβIIIa.","Antagonism of platelet αIIbβIIIa (also known as the fibrinogen receptor) is known to block platelet aggregation in humans.","In order to avoid bleeding side-effects when treating the conditions of disease states associated with the integrin αvβ3, it would be beneficial to develop agents which selectively bind to αvβ3 as opposed to αIIbβIIIa as well as other integrins.","Polypeptides or polypeptide monobodies selective for αvβ3 offer such an advantage.","As noted above, integrins are believed to be involved in, among other biological processes, tumor metastasis and neoplasia.","Tumor cell invasion occurs by a three step process: 1) tumor cell attachment to extracellular matrix; 2) proteolytic dissolution of the matrix; and 3) movement of the cells through the dissolved barrier.","This process can occur repeatedly and can result in metastases at sites distant from the original tumor.","Studies have shown (Humphries et al., 1988), that RGD-like compounds can interfere with experimental metastasis wherein tumor cells are injected directly into the blood.","In addition, Seftor et al.","(1992) have shown that the αvβ3 integrin has a biological function in melanoma cell invasion.","Montgomery et al., (1994) demonstrated that the integrin αvβ3 expressed on human melanoma cells promotes a survival signal, protecting the cells from apoptosis.","Accordingly, mediation of the tumor cell metastatic pathway by interference with the αvβ3 integrin cell adhesion receptor to impede tumor metastasis would be beneficial.","Brooks et al.","(1994a) have demonstrated that antagonists of αvβ3 provide a therapeutic approach for the treatment of neoplasia (inhibition of solid tumor growth) since systemic administration of αvβ3 antagonists causes dramatic regression of various histologically distinct human tumors.","The adhesion receptor integrin αvβ3 was also identified as a marker of angiogenic blood vessels in chick and man and plays a critical role in angiogenesis or neovascularization (Brooks et al., 1995).","Angiogenesis is characterized by the invasion, migration and proliferation of smooth muscle and endothelial cells.","Antagonists of αvβ3 inhibit this process by selectively promoting apoptosis of cells in neovasculature.","The growth of new blood vessels, or angiogenesis, also contributes to pathological conditions such as diabetic retinopathy (Adonis et al., 1994), rheumatoid arthritis (Peacock et al., 1992) and osteoarthritis (Ondrick et al., 1992).","Therefore, αvβ3 antagonists are useful targets for treating or preventing such conditions associated with neovascularization (Brooks et al., 1994b).","Because angiogenesis occurs normally in the female reproductive organs, antagonists of αvβ3 would be useful in controlling fertility.","It has also been reported that the cell surface receptor αvβ3 is the major integrin on osteoclasts responsible for attachment to bone.","Osteoclasts cause bone resorption and when such bone resorbing activity exceeds bone forming activity it results in osteoporosis (a loss of bone), which leads to an increased number of bone fractures, incapacitation and increased mortality.","Antagonists of αvβ3 have been shown to be potent inhibitors of osteoclastic activity both in vitro (Sato et al., 1990) and in vivo (Fisher et al., 1993).","Antagonism of αvβ3 leads to decreased bone resorption and therefore restores a normal balance of bone forming and resorbing activity.","Thus, it would be beneficial to provide antagonists of osteoclast αvβ3 which are effective inhibitors of bone resorption and, therefore, are useful in the treatment or prevention of osteoporosis.","The role of the αvβ3 integrin in smooth muscle cell migration also makes it a therapeutic target for prevention or inhibition of neointimal hyperplasia which is a leading cause of restenosis after vascular procedures (Choi et al., 1994).","Prevention or inhibition of neointimal hyperplasia (restenosis) by the polypeptides or polypeptide monobodies of the present invention will be beneficial.","White (1993) has reported that adenovirus uses αvβ3 for entering host cells.","The integrin appears to be required for endocytosis of the virus particle and may be required for penetration of the viral genome into the host cell.","Thus, polypeptides or polypeptide monobodies of the present invention which inhibit αvβ3 will be useful as antiviral agents.","As a result of the present invention, polypeptides or polypeptide monobodies of the present invention (with αvβ3 selective binding activity) can be used for blocking αvβ3-mediated events such as cell adhesion, osteoclast-mediated bone resorption, restenosis, ocular neovascularization and growth of hemangiomas, as well as neoplastic cell or tumor growth and dissemination.","The inventive polypeptide or polypeptide monobodies can also be used for mediated targeting and delivery of therapeutics for disrupting or killing αvβ3 bearing neoplasms and tumor-related vascular beds.","In addition, the inventive polypeptide or polypeptide monobodies can be used for visualization or imaging of αvβ3 bearing neoplasms or tumor related vascular beds, for example, by MRI or immunoscintigraphy.","In addition, these polypeptide or polypeptide monobodies can be used to detect αvβ3 in solution, in frozen tissue sections, and on the surface of cells; therefore, these polypeptides or polypeptide monobodies may be used for the detection and characterization of αvβ3-bearing tumor and endothelial cells in human malignancies.","Accordingly, the polypeptides or polypeptide monobodies of the present invention may be used for identification of tumor vasculature.","Since the integrin αvβ3 is minimally expressed on resting or normal blood vessels, but is significantly up-regulated on vascular cells within human tumors (Brooks et al., 1994a; Brooks et al., 1994b; Brooks et al., 1995), αvβ3 may be considered a marker of human tumor-associated blood vessels and tumor growth (Brooks et al., 1995).","Consequently, polypeptides or polypeptide monobodies that recognize αvβ3 may be used as a diagnostic agent to identify tumor-related blood vessels or vascular beds using conventional immunohistochemical techniques.","Further, since the integrin αvβ3 is a marker of tumor-associated blood vessels and recent findings indicate that blood vessel density is a prognostic indicator of cancer and disease status (Weidner, 1993), polypeptides or polypeptide monobodies that recognize αvβ3 may be used as a component of a diagnostic device or technique to measure tumor-related blood vessel abundance or density and to define disease prognosis.","Specifically, histologic sections from fresh-frozen or formalin-fixed paraffin embedded tumor tissue are immunostained with αvβ3-specific polypeptides or polypeptide monobodies, i.e., at concentration of 0.1-50 μg/ml, using techniques and procedures known to those skilled in the art.","αvβ3-specific polypeptides or polypeptide monobodies bound to vascular endothelial cells in the tissue can be visualized using a polypeptide or polypeptide monobody fusion or a secondary agent that permits localization of the αvβ3-specific polypeptides or polypeptide monobodies bound within the tissue section.","For fusions, the αvβ3-specific polypeptides or polypeptide monobodies can be fused with alkaline phosphatase, both of which retain their functionality.","The alkaline phosphatase, which is integral with the functional polypeptide or polypeptide monobody portion, can be identified in accordance with known procedures.","The secondary agent is typically an antibody that reacts or binds with the αvβ3-specific polypeptides or polypeptide monobodies and has an attached reporter molecule, e.g., an anti-polypeptide or polypeptide monobody antibody with an attached enzyme or fluorescent marker.","The polypeptide or polypeptide monobody, when used with secondary agents, is typically provided with an epitope tag fusion, the epitope portion being recognized by the anti-polypeptide or polypeptide monobody antibody.","The number and area of immunostained blood vessels in the tissue can then be counted by microscopic techniques, familiar to those skilled in the art, to assess tumor blood vessel density or abundance.","The fusion proteins of the present invention which include a DNA binding polypeptide (e.g., polylysine amino acid sequence) can also be used to deliver a therapeutic DNA molecule into a cell which expresses an integrin targeted by the polypeptide or polypeptide monobody of the present invention, such as the αvβ3 integrin.","Polylysine has long been known as a DNA-binding motif because of its role in histone-mediated DNA condensation.","This property of polylysine to bind DNA was utilized to deliver the DNA to mammalian cells (Wu & Wu, 1987; Wu & Wu, 1988).","The technology was utilized by Zhou et al to develop lipophilic polylysines which were in part the basis of present lipofection delivery of DNA (Zhou et al., 1991).","Polylysines conjugated by peptide bond to integrin-binding peptides have been described which effectively deliver DNA to cells (Harbottle et al., 1998).","Polylysine DNA has also been used to deliver DNA to endothielial cells, a potential target for αvβ3-selective gene delivery (Trubetskoy et al., 1992).","Thus, a fusion protein, as described above, is first combined with the DNA to be delivered under conditions which are effective to allow the DNA binding peptide, such as polylysine, to bind the DNA.","Thereafter, the complex formed upon binding of the fusion protein to the DNA, is administered to a patient under conditions effective to deliver the DNA into the cells which express the integrin, e.g., αvβ3, targeted by the fusion protein.","In addition to its administration alone, a number of known delivery techniques can be utilized for the delivery, into cells, of either the polypeptide monobodies themselves or nucleic acid molecules which encode them.","Regardless of the particular method of the present invention which is practiced, when it is desirable to contact a cell (i.e., to be treated) with a polypeptide or polypeptide monobody of the present invention, or its encoding nucleic acid, it is preferred that the contacting be carried out by delivery of the polypeptide monobody or its encoding nucleic acid to the cell.","One approach for delivering the polypeptide or polypeptide monobody to the cell or its encoding nucleic acid into cells involves the use of liposomes.","Basically, this involves providing the polypeptide or polypeptide monobody or its encoding nucleic acid to be delivered, and then contacting the target cell with the liposome under conditions effective for delivery of the polypeptide or polypeptide monobody or nucleic acid into the cell.","Liposomes are vesicles comprised of one or more concentrically ordered lipid bilayers which encapsulate an aqueous phase.","They are normally not leaky, but can become leaky if a hole or pore occurs in the membrane, if the membrane is dissolved or degrades, or if the membrane temperature is increased to the phase transition temperature.","Current methods of drug delivery via liposomes require that the liposome carrier ultimately become permeable and release the encapsulated drug at the target site.","This can be accomplished, for example, in a passive manner wherein the liposome bilayer degrades over time through the action of various agents in the body.","Every liposome composition will have a characteristic half-life in the circulation or at other sites in the body and, thus, by controlling the half-life of the liposome composition, the rate at which the bilayer degrades can be somewhat regulated.","In contrast to passive drug release, active drug release involves using an agent to induce a permeability change in the liposome vesicle.","Liposome membranes can be constructed so that they become destabilized when the environment becomes acidic near the liposome membrane (Wang & Huang, 1987).","When liposomes are endocytosed by a target cell, for example, they can be routed to acidic endosomes which will destabilize the liposome and result in drug release.","Alternatively, the liposome membrane can be chemically modified such that an enzyme is placed as a coating on the membrane which slowly destabilizes the liposome.","Since control of drug release depends on the concentration of enzyme initially placed in the membrane, there is no real effective way to modulate or alter drug release to achieve “on demand” drug delivery.","The same problem exists for pH-sensitive liposomes in that as soon as the liposome vesicle comes into contact with a target cell, it will be engulfed and a drop in pH will lead to drug release.","This liposome delivery system can also be made to accumulate at a target organ, tissue, or cell via active targeting (e.g., by incorporating an antibody or hormone on the surface of the liposomal vehicle).","This can be achieved according to known methods.","Different types of liposomes can be prepared according to Bangham et al.","(1965); U.S. Pat.","No.","5,653,996 to Hsu et al.","; U.S. Pat.","No.","5,643,599 to Lee et al.","; U.S. Pat.","No.","5,885,613 to Holland et al.","; U.S. Pat.","No.","5,631,237 to Dzau et al.","; and U.S. Pat.","No.","5,059,421 to Loughrey et al., as well as any other approach demonstrated in the art.","From the foregoing, it should be apparent that the polypeptide or polypeptide monobody can either be delivered in a liposome or, when incorporated into the lipid bilayer(s), used as a means for targeting a liposome to αvβ3 integrin-expressing cells.","An alternative approach for delivery of polypeptide monobodies involves the conjugation of the desired polypeptide or polypeptide monobody to a polymer that is stabilized to avoid enzymatic degradation of the conjugated polypeptide or polypeptide monobody.","Conjugated proteins or polypeptides of this type are described in U.S. Pat.","No.","5,681,811 to Ekwuribe.","When it is desirable to achieve heterologous expression of a desirable polypeptide or polypeptide monobody of the present invention in a target cell, DNA molecules encoding the polypeptide or polypeptide monobody can be delivered into the cell.","Basically, this includes providing a nucleic acid molecule encoding the polypeptide or polypeptide monobody and then introducing the nucleic acid molecule into the cell under conditions effective to express the polypeptide monobody in the cell.","Preferably, this is achieved by inserting the nucleic acid molecule into an expression vector before it is introduced into the cell.","For intracellular expression in this manner, use of polypeptide monobodies is preferred because such monobodies are enzymatically stable within cells.","When transforming mammalian cells for heterologous expression of a polypeptide or polypeptide monobody of the present invention, an adenovirus vector 5 can be employed.","Adenovirus gene delivery vehicles can be readily prepared and utilized given the disclosure provided in Berkner (1988) and Rosenfeld et al.","(1991).","Adeno-associated viral gene delivery vehicles can be constructed and used to deliver a gene to cells.","The use of adeno-associated viral gene delivery vehicles in vivo is described in Flotte et al.","(1993) and Kaplitt et al.","(1994).","Additional types of adenovirus vectors are described in U.S. Pat.","No.","6,057,155 to Wickham et al.","; U.S. Pat.","No.","6,033,908 to Bout et al.","; U.S. Pat.","No.","6,001,557 to Wilson et al.","; U.S. Pat.","No.","5,994,132 to Chamberlain et al.","; U.S. Pat.","No.","5,981,225 to Kochanek et al.","; U.S. Pat.","No.","5,885,808 to Spooner et al.","; and U.S. Pat.","No.","5,871,727 to Curiel.","Retroviral vectors which have been modified to form infective transformation systems can also be used to deliver nucleic acid encoding a desired polypeptide monobody into a target cell.","One such type of retroviral vector is disclosed in U.S. Pat.","No.","5,849,586 to Kriegler et al.","Regardless of the type of infective transformation system employed, it should be targeted for delivery of the nucleic acid to a specific cell type.","For example, for delivery of the nucleic acid into tumor cells, a high titer of the infective transformation system can be injected directly within the tumor site so as to enhance the likelihood of tumor cell infection.","The infected cells will then express the desired polypeptide or polypeptide monobody, allowing the polypeptide or polypeptide monobody to bind to the αvβ3 integrin.","Whether the polypeptides or polypeptide monobodies of the present invention (or encoding nucleic acids) are administered alone or in combination with pharmaceutically or physiologically acceptable carriers, excipients, or stabilizers, or in solid or liquid form such as, tablets, capsules, powders, solutions, suspensions, or emulsions, they can be administered orally, parenterally, subcutaneously, intravenously, intramuscularly, intraperitoneally, by intranasal instillation, by intracavitary or intravesical instillation, intraocularly, intraarterially, intralesionally, or by application to mucous membranes, such as, that of the nose, throat, and bronchial tubes.","For most therapeutic purposes, the polypeptides or polypeptide monobodies (or encoding nucleic acids) can be administered intravenously.","For injectable dosages, solutions or suspensions of these materials can be prepared in a physiologically acceptable diluent with a pharmaceutical carrier.","Such carriers include sterile liquids, such as water and oils, with or without the addition of a surfactant and other pharmaceutically and physiologically acceptable carrier, including adjuvants, excipients or stabilizers.","Illustrative oils are those of petroleum, animal, vegetable, or synthetic origin, for example, peanut oil, soybean oil, or mineral oil.","In general, water, saline, aqueous dextrose and related sugar solution, and glycols, such as propylene glycol or polyethylene glycol, are preferred liquid carriers, particularly for injectable solutions.","For use as aerosols, the polypeptide monobodies or nucleic acids in solution or suspension may be packaged in a pressurized aerosol container together with suitable propellants, for example, hydrocarbon propellants like propane, butane, or isobutane with conventional adjuvants.","The materials of the present invention also may be administered in a non-pressurized form such as in a nebulizer or atomizer.","Dosages to be administered can be determined according to known procedures, including those which balance both drug efficacy and degree of side effects.","From the foregoing, it should be appreciated that therapeutic methods and compositions are contemplated for inhibiting integrins, particularly αvβ3 integrin.","These methods are useful to inhibit cell attachment and migration mediated by the integrins and find application in a wide variety of cell types, tissues and systems where attachment of cells is not desired.","Thus, in general, the invention contemplates a method for inhibiting an integrin that binds a specific ligand by contacting the integrin with an inhibiting amount of an integrin-inhibiting polypeptide or polypeptide monobody (or fusion protein/conjugate containing them) of the present invention.","Typically, the method is practiced on cells expressing the integrin on the surface of the cell, so the contacting occurs by exposing the cells to the integrin-inhibiting polypeptides or polypeptide monobodies.","An inhibiting amount of integrin-inhibiting polypeptide or polypeptide monobody is an amount sufficient to produce the desired result, namely to inhibit the integrin to a degree sufficient to reduce the adhesion of the cell expressing the integrin, and typically depends on the amount of integrin to be contacted.","In preferred embodiments, whether the method is practiced in vitro or in vivo, an inhibiting amount is an amount sufficient to provide at least about one molar equivalent of integrin-inhibiting polypeptide or polypeptide monobody per molar equivalent of integrin to be inhibited.","This amount is referred to as a stoichiometric amount of integrin-inhibiting polypeptide or polypeptide monobody.","Although polypeptide or polypeptide monobody affinity is typically sufficient for an integrin-inhibiting polypeptide or polypeptide monobody to react stoichiometrically in dilute solutions, it is preferred that an inhibiting amount is in the range of about 100 nanomolar (nM) to 1 millimolar (mM), preferably in the range of 1 to 100 micromolar (μM), more preferably about 1 to 10 μM.","When an integrin-inhibiting method is practiced in vitro, a liquid sample containing integrin, and preferably a physiological fluid containing cells that express cell surface integrin, are admixed with an inhibiting amount of an integrin-inhibiting polypeptide or polypeptide monobody of the present invention to form a complex.","The complex is maintained under biological conditions compatible with the formation of an reaction product and also compatible, if required, with cell viability for a time period sufficient for the integrin-inhibiting polypeptide or polypeptide monobody to react with the integrin and, when reacted, inhibit the integrin.","When the integrin-inhibiting method is practiced in vivo, an inhibiting amount of a polypeptide or polypeptide monobody composition containing a physiologically tolerable diluent and integrin-inhibiting polypeptide or polypeptide monobody molecules as described herein is administered to a mammal, e.g.","a human, and the mammal is maintained for a sufficient period to allow the monobody molecules to react with any integrin present and form an inhibiting reaction product.","Suitable routes of administration are envisioned as described above, preferably intravenous, intraperitoneal, intra muscular, intrathecal or subcutaneous.","The integrin-inhibiting polypeptide or polypeptide monobody may be combined with other pharmaceutical compositions and/or excipients.","For example, αvβ3 integrin-inhibiting polypeptide or polypeptide monobody may be co-administered or added to established anti-cancer chemotherapeutic or biotherapeutic regimens.","Normal physiologic saline is a preferred excipient or vehicle for administration of the polypeptide or polypeptide monobody.","This may include, but is not limited to, combining or co-administering αvβ3 integrin-inhibiting polypeptide or polypeptide monobody with cytotoxic drugs, combinations of cytotoxic drugs or with immune stimulating drugs, such as interleukins or their derivatives or with hematopoietic factors and their derivatives.","For example, αvβ3 monobody may be co-administered or added to therapeutic regimens for the use IL-1, IL-2, IL-4, IL-6, IL-12, IL-15, TNF, α, β, γ, interferons and M-CSF, or combinations of these agents or their derivatives, in biologic therapy of cancer.","Further, αvβ3 integrin-inhibiting polypeptide or polypeptide monobody may be co-administered or added to therapeutic regimens for the use of G-CSF, M-CSF, IL-3 and erythropoietin, or combinations of these agents or their derivatives, in biologic therapy of cancer.","In addition to its use alone, the integrin-inhibiting polypeptide or polypeptide monobody may also be conjugated to a chemotherapeutic agent to afford a conjugate that can be administered alone or in combination with other anti-cancer drugs or immune-enhancing drugs of the type described above.","Conjugates can be formed using known techniques, although typically the conjugate can be formed by linking the chemotherapeutic agent directly or indirectly, via covalent bond, to the C-terminal or N-terminal regions of the monobody.","For purposes of imaging cancerous or pre-cancerous lesions to assess the degree of angiogenesis and, therefore, the likelihood of metastatic activity, the polypeptide or polypeptide monobody of the present invention can be formed as a fusion protein including a metal ion-complexing polypeptide or formed as a conjugate including a contrasting agent or chelating agent.","Suitable metal ion-complexing polypeptides include, without limitation, polyhistidine.","The fusion protein-metal ion complex can be used to identify integrin containing tissues, particularly αvβ3 integrin containing tissues, i.e., where angiogenesis or neoplasia may be occurring.","The complexation of the metal ions to the fusion protein, and specifically to the metal ion-complexing polypeptide or chelating agent, is achieved by mixing appropriate amounts of the fusion protein with the metal ion.","This is preferably done in solution, with the solution including an appropriate buffer.","In one approach, the metal ion is, when mixed with the fusion protein, already in the oxidation state required for complexing to the metal ion-complexing polypeptide.","Some metal ions are complexed in their most stable oxidation state, such as ionic forms of calcium, potassium, indium, manganese, copper, zinc, cobalt and other metals.","In another approach, the metal ions must be reduced to a lower oxidation state in order to be complexed to the metal ion-complexing polypeptide.","This is true of ferrous, ferric, stannous, stannic, technetiumoxo[V], pertechnetate, rheniumoxo[V], perrhenate and other similar metal ions.","Thus, for example, both perrhenate and pertechnetate must be reduced to a lower oxidation state prior to complexing.","Reduction may be performed prior to mixing with the polypeptide, simultaneously with mixing with the fusion protein, or subsequent to mixing with the fusion protein.","Any means of reduction of metal ions to the desired oxidation state known to the art may be employed.","For example, perrhenate or pertechnetate may be reduced by use of stannous ions, dithionite or other means.","The stannous or dithionite metal ion reducing agent may be mixed with the metal ion to be reduced either prior or subsequent to addition of the metal ion to the fusion protein, or the reducing agent may be in solution with the fusion protein when the metal ion to be reduced, and subsequently complexed to the fusion protein, is added to the solution.","The stoichiometric ratios between the fusion protein and the metal ion in the labeling or complexing step can be varied depending on the application.","For example, in the case of radiometal complexation, the ratio of radiometal ions to fusion protein molecules can be varied from less than 1:2 to 1:1000 or higher, without generating substantial radiochemical impurities.","In other applications, the ratio of metal ions to fusion protein molecules can range from at least 1000:1 to 1:1000 or higher.","When the concentration of metal ions is higher than the concentration of fusion protein molecules, all or virtually all of the fusion protein molecules will be complexed to metal ions.","Either radioactive or non-radioactive metal ions can be employed.","Where a specific diagnostic or therapeutic advantage can be obtained from use of a radioisotope, then a radioactive metal ion is employed.","Where the diagnostic or therapeutic utility is obtained from the conformationally constrained biological-function domain, a non-radioactive metal ion is employed.","It is possible and contemplated, and may for certain applications be advantageous, to employ different metal ions with the same conjugate product of this invention.","Suitable contrasting agents include, without limitation, a dextran-coated superparamagnetic iron oxide (SPIO) particle (Reimer et al., 1991; Schaffer et al., 1993; Reimer et al., 1994; Small et al., 1994; Kresse et al., 1998; Suwa et al., 1998), a lipid vesicle or other similar particle with Gd3+ attached to the surface (Kabalka et al., 1991; Tilcock et al., 1992; Sipkins et al., 1998), and poly-lysine with Gd3+ attached to the amino groups (Kayyem et al., 1995; Vera et al., 1995).","Like the fusion proteins containing a metal ion complexing agent, the conjugates containing a contrasting agent can also be used to identify integrin containing tissues, particularly αvβ3 integrin containing tissues, i.e., where angiogenesis or neoplasia may be occurring.","Another type of conjugate includes an organic chelating agent (e.g., diethylenetriamine pentaacetic acid) conjugated to a polypeptide monobody of the present invention which has been modified at, e.g., its C-terminus to include a cysteine residue which will form a stable covalent bond with the chelating agent.","Thereafter, the conjugate can be exposed to a suitable metal ion of the type described above in a manner which is effective to allow the conjugated diethylenetriamine pentaacetic acid moiety to complex with the metal ions.","The fusion proteins or conjugates, and the methods of this invention can also be applied to diagnostic agents for use in positron emission tomography (PET) and magnetic resonance imaging (MRI).","For use as a PET agent, a fusion protein or conjugate is complexed with one of the various positron emitting metal ions, such as 51Mn, 52Fe, 60Cu, 68Ga, 72As, 94mTc, 110In, and isotopes of At.","For MRI applications, the complexing metal ion is paramagnetic, such as Mn, Gd, Fe, or Dy.","EXAMPLES The following Examples are intended to be illustrative and in no way are intended to limit the scope of the present invention.","Materials & Methods The following materials and methods were used in the experiments set forth in Examples 1-5.General reagents: All chemicals, unless otherwise specified, were obtained from Sigma (St. Louis, Mo.).","Purified integrins, cell adhesion strips, and LM609 were purchased from Chemicon (Temecula, Calif.).","Echistatin and GRGDSPK peptide (SEQ ID No: 26) were obtained from Bachem (Torrence, Calif.).","Cell Culture: K562, K562-αvβ3, K562-αIIbβ3, K562-αvβ5, K562-α4β1, and K562-α4β7 cells were a gift of Dr. S. Blystone (Upstate Medical University, Syracuse, N.Y.) (Blystone et al., 1994).","K562 cells were maintained in Iscove's Modified Dulbecco's Medium (IMDM) (Gibco, Invitrogen, Grand Island, N.Y.) supplemented with 10% fetal bovine serum (Sigma), 0.5 U/L penicillin-streptomycin, and 2 mM L-Glutamine (Gibco).","K562-αvβ3, K562-αIIbβ3, K562-αvβ5, K562-α4β1, and K562-α4β7 cells were maintained in the same media containing 500 μg/ml G418 (GENETICIN) (Gibco).","Construction of phage vectors: A phage-display vector for FNfn10, designated JCFN, was constructed by cloning the FNfn10 gene (Koide et al., 1998) in the modified M13 vector JC-M13-88 (Chappel et al., 1998).","The FNfn10 gene was amplified from pAS38 (Koide et al., 1998) using the following oligonucleotides: FN3JCM8Nhe (SEQ ID No:29) cctagctagc gtagctcagg ccatgcaggt ttctgatgtt c 41 and FN3JCM8Hin (SEQ ID No:30) ggccaagctt gaccgccacc agaaccgcca ccggtacggt ag 42 digested with the restriction enzymes NheI and HindIII, respectively, and subcloned in JC-M13-88 using the same restriction enzyme sites so that the FNfn10 gene is placed in-frame between the OmpA signal sequence and M13 geneVIII.","It was confirmed by ELISA that the JCFN phages prepared using E. coli XL-1blue displayed FNfn10 on the phage surface.","Purified FNfn10 clones were expressed with an N-terminal histag and a C-terminal GKKGK (SEQ ID No: 31) tag.","The GKKGK (SEQ ID No: 31) tag was incorporated to increase the solubility (Koide et al., 1998) and to serve as a preferred biotinylation site.","FNfn10 genes were amplified using the following oligonucleotides: FN1F2 (SEQ ID No:32) cgggatccca tatgcaggtt tctgatgttc cgcgtgacct ggaagttgtt gctgcgacc 59 and FN3GKKGK (SEQ ID No:33) ccgactcgag ttactattta ccttttttac cggtacggta gttaatcgag 50 which were digested with the restriction enzymes NdeI and XhoI, and ligated with pAS45 (Koide et al., 1998) digested with the same restriction enzymes.","The expression vector for the wild type protein, pAS54, contained an Arg6 to Thr mutation that had been introduced to remove a secondary Thrombin site (Koide et al., 1998).","Construction of the FNfn10 library: A FNfn10 library, JCFN-RGD, was constructed in such a way that the FG loop has an XRGDXXXX sequence where X stands for any amino acids (residue #77-84; residue numbering is according to FIG.","2(a) of Koide et al.","(1998)).","Mutagenesis on the JCFN template was performed according to Kunkel's method (Kunkel, 1985) using an oligonucleotide, designated JCFNFGRGD, as follows: JCFNFGRGD (SEQ ID No: 34) gttaatcgag attggcttgg arnnnmnnmnn mnnatcgccg cgmnnagtaa cagcgtatac 60 where N is a mixture of A, G, C and T, and M is a mixture of A and C).","E. coli SS320 was electro-transformed with the mutagenesis mixture, resulting in a phage library containing 1.5×109 independent clones.","The phages were re-amplified in E. coli XL1-blue with various concentrations of isopropyl β-D-thiogalactopyranoside (IPTG).","αvβ3 biopanning using the JCFN-RGD library: Integrin biopanning was performed similarly to the method previously described (Koivunen et al., 1994).","Purified αvβ3 (5 μg/ml in TBS buffer containing 2 mM CaCl2) was bound to a single microtiter well of a 96-well plate.","The well was rinsed 3 times with sterile water containing 2 mM CaCl2 and blocked with 5% phage blocking reagent (Novagen, Darmstadt, Germany) for one hour at room temperature.","The plate was rinsed 5 times with TBS buffer containing 0.1% Tween-20 and 2 mM CaCl2, then incubated with JCFN-RGD library diluted in 5% blocking reagent for one hour.","The plate was washed ten times with TBS buffer containing 0.1% Tween 20 and 2 mM CaCl2 and bound phage were eluted with 0.2 M glycine-HCl (pH 2.2) containing 1 mg/ml BSA for 10 minutes at room temperature with gentle agitation and neutralized with 15% (v/v) of 1 M Tris-HCl (pH 9.1).","The phage were amplified using the E. coli ER2738 host strain (New England Biolabs, Beverly, Mass.)","in the presence of 1 mM, 0.1 mM or with no IPTG and purified by precipitating with 20% (w/v) polyethylene glycol-8000, 2.5 M NaCl and resuspending in PBS containing 20% (v/v) glycerol.","The biopanning and amplification of eluted phage were repeated for two more rounds.","Upon completion of three rounds of biopanning, individual phage clones were amplified and the genomes were purified by phenol extraction.","Polymerase chain reaction was performed to amplify the FNfn10 domain using the following primers: SFN3F (SEQ ID No:35) agctcaattg gtccggtgga ggttctgatg ttccgcgtga cctg 44 and SFN3R (SEQ ID No:36) agctaagctt ttaggtacgg tagttaatcg agat 34 The DNA was also sequenced using the SFN3F primer.","Protein preparation: The expression of FNfn10 proteins was performed as previously described (Koide et al., 1998) except that the proteins were further biotinylated.","After biotinylation, the protein-containing solution was applied to a nickel affinity column and unbound materials were washed off; the column was then equilibrated with 50 mM sodium phosphate buffer (pH 8.0) containing 500 mM sodium chloride.","D-biotinoyl-ε-aminocaptoic acid N-hydroxysuccinimide ester (726 μM) (Boehringer Mannheim, Mannheim, Germany) in the same buffer was applied to the column, and the column was incubated for an hour at room temperature to perform biotinylation.","This biotinylation process was performed a total of three times.","Then the column was washed with 20 mM Tris-HCl buffer (pH 8.0) containing 500 mM sodium chloride, and the biotinylated FNfn10 was eluted with the buffer containing 500 mM imidazole.","The proteins purified in this manner were >90% pure as judged by SDS-PAGE and reverse phase chromatography.","Protein concentrations for subsequent analyses were determined by Bradford assay and confirmed by SDS-PAGE analysis and Coomassie Brilliant Blue staining.","Biotinylation was confirmed by Western analysis with streptavidin-horseradish peroxidase (Oncogene, Cambridge, Mass.).","ELISA detection of biotinylated FNfn10 clones: Purified integrin was bound to a flat-bottom, high-binding EIA/RIA plate (Corning Costar, Corning, N.Y.) at a concentration of 5 μg/ml in TBS for 12-16 hours at 4° C. The plate was washed once with TBS and blocked for 2 hours at room temperature with 0.1M NaHCO3, 0.5 mg/ml BSA, 0.2% NaN3, pH 8.6.Biotinylated FNfn10 clones were bound in TBST+Ca2+ (TBS 0.1% Tween-20, 2 mM CaCl2).","Plates were washed ten times with TBST+Ca2+ and incubated 20 minutes with 2 μg/ml streptavidin-horseradish peroxidase (Oncogene) in TBST+Ca2+ at room temperature.","Plates were washed ten times with TBST+Ca2+.","Bound peroxidase was detected with ABTS (2,2′-azino-di-3-ethyl-benzthiazoline sulfonic acid) peroxidase substrate (Sigma), 1 mg/ml in 0.1 M NaCitrate, 0.1 M Na2HPO4, pH 4.0 supplemented with 0.03% hydrogen peroxide.","Color was allowed to develop for 10-15 minutes and the A400 was read with a SpectraCount ELISA reader (Packard, Downers Grove, Ill.).","Flow cytometric detection of cell-binding by biotinylated FNfn10 & FNfn10-3JCLI4: K562 cells (105) transfected with various integrins were resuspended in 5 ml FACS buffer (PBS, 0.5% BSA, 0.1% NaN3), washed, and resuspended in FACS buffer containing either 0.4 μg/ml FNfn10 protein (wild-type of 3JCLI4) or 1 μg/ml LM609, prior to incubation at room temperature for 20 minutes.","Cells were then washed with 5 ml FACS buffer, incubated in 1:50 streptavidin-APC (BD Pharmingen, San Diego, Calif.) for 20 minutes and washed again with 5 ml FACS buffer, prior to detection of cell surface fluorescence using a FACSCalibur flow cytometer (Becton Dickinson, San Jose, Calif.).","Cell adhesion assays: Fibronectin and vitronectin-coated strips (Chemicon) were rehydrated 15 minutes in PBS.","Inhibitor in IMDM/10% FBS was added to the bottom of the well, and 105 cells/well in maintenance media were added to a final volume of 100 μl/well.","Cells were incubated 2 hrs at 37° C., 5% CO2, washed three times with PBS at room temperature, and stained 10 minutes with 0.2% crystal violet, 10% ethanol.","Cells were then washed three times with PBS and solubilized with a 1:1 ratio of PBS:absolute ethanol for 10 minutes with agitation.","A600 was measured using a SpectraCount ELISA reader (Packard).","Example 1 Screening of the JCFN-RGD Phage Display Library for αvβ3 Integrin Binding In the FNfn10 structure of SEQ ID No: 2 (wild-type), the FG loop corresponds to residues 78-87, and the RGD sequence to residues 79-81 (residue numbering according to the Protein Data Bank entry 1TTG, Main et al., 1992).","Five residues immediately adjacent to the RGD sequence were modified in the FG loop using Kunkel mutagenesis as described above.","The JCFN-RGD (XRGDXXXX, SEQ ID No: 4) library contained 1.5×109 independent clones, sufficiently large to include all possible sequences.","The library was amplified in 0.0, 0.1, or 1.0 mM IPTG to vary FNfn10 copy number and screened for binding to αvβ3 integrin.","After 3 rounds of biopanning, 100-1000 fold increased recovery of phage was observed compared to the initial library.","Of 20 phage clones sequenced, 8 contained an XRGDWXEX (SEQ ID No: 26) consensus sequence (FIG.","4A).","Phage inputs of 109, 108, or 107 pfu were then allowed to bind to αvβ3 or BSA coated wells and bound phages were recovered with 0.2M glycine, pH 2.2.Using phage clones amplified in 1 mM IPTG (high FNfn10 copy number), phage clones containing the consensus were recovered at levels of 0.01-1% input from αvβ3 coated wells; the clones were recovered at approximately 1000-fold lower levels from BSA coated wells and integrin-binding was found to be calcium-dependent.","In contrast, clones which lacked the XRGDWXEX (SEQ ID No: 26) motif were recovered at equivalent levels from both αvβ3 coated and BSA coated wells.","The RGDWXE-containing clones were amplified in the absence of IPTG so as to decrease the expressed copy number of FNfn10 protein on the phage surface, and the panel of clones were then directly compared in a phage binding assay, for their ability to bind to αvβ3 (FIG.","4B).","The clones recovered at the highest level were 2JCAV1, 2JCAV2, 3JCAV3, 3JCNI1, and 3JCLI4, all of which were recovered at a level of approximately 0.1% of input phage from αvβ3-coated wells and at 1000-fold lower levels from BSA-coated wells.","Therefore, varying FNfn10 copy number on M13 by IPTG concentration did not have an effect on the relative binding efficiency of the clones isolated, suggesting that the binding interaction between the αvβ3 integrin and the phage-displayed FNfn10 derivatives might be of high affinity.","The modified FNfn10 proteins expressed by this panel of phage clones were expressed as an isolated protein in E. coli, purified and biotinylated.","3JCLI4 (the second-highest recovered clone in phage binding assays), was selected for further study because the purified FNfn10 protein derived from clone 2JCAV1 (the highest recovered clone) proved to be rather insoluble and prone to precipitation.","SDS polyacrylamide gel electrophoresis and Coomassie blue staining revealed the presence of a single protein of the expected size (˜15 kDa) within the purified preparation of biotinylated FNfn10-3JCLI4 protein.","Western blot analysis using a monoclonal antibody specific for the hexahistidine epitope tag on the purified FNfn10 protein confirmed the identity of this protein, and further analysis revealed that the binding of both the phage clone 3JCLI4 and of the corresponding purified protein (FNfn10-3JCLI4) to αvβ3 integrin was Ca++ dependent and could be eliminated in the presence of EDTA.","Example 2 Determining Affinity and Specificity of FNfn10-3JCLI4 for αvβ3 Integrin Biotinylated FNfn10-3JCLI4 and biotinylated wild-type FNfn10 (FNfn10-WT) were allowed to bind to αvβ3, α1β1, or BSA coated wells in approximate half-log increments from 0.0001 to 0.1 μg/ml, corresponding to molar concentrations of 8 pM to 8 nM.","Bound biotinylated protein was detected by ELISA (FIG.","5A).","Assuming a high ratio of free to bound FNfn10-3JCLI4, the equation y=(m1*x/(m2+x)) was fit to the data (R>0.99) to determine the half-maximal binding affinity of 0.01 μg/ml or 800 pM.","At each concentration examined, the binding of biotinylated FNfn10-3JCLI4 to αvβ3 was higher than that for FNfn10-WT, and neither protein was found to bind detectably to the α1β1 integrin (i.e., binding to α1β1 was identical to binding to BSA).","Next, the kinetics of protein association and dissociation was examined.","After allowing biotinylated FNfn10-3JCLI4 to bind to αvβ3-coated wells, 100 fold excess unlabelled FNfn10-3JCLI4 was added to some wells and the plates were incubated for a further 24 hours; control wells received were incubated in the absence of the unlabelled competitor.","At the end of this time, the amount of plate-bound biotinylated FNfn10-3JCLI4 was assessed by ELISA assay.","No difference was detected between wells that were incubated for 24 hours in the presence or absence of excess unlabelled competitor protein.","This suggests that the rate of dissociation of FNfn10-3JCLI4 from αvβ3 is very slow (>24 hours; FIG.","5B).","To examine the rate of association between αvβ3 and biotinylated 3JCLI4, biotinylated FNfn10-3JCLI4, at concentrations of 0.01, 0.02, or 0.05 μg/ml, was allowed to bind to αvβ3-coated wells for periods of time ranging from one minute to two hours.","For all three concentrations, levels of bound FNfn10-3JCLI4 were detectable within one minute and reached saturation levels within two hours (FIG.","5C).","Therefore, in light of the prolonged dissociation rate revealed in FIG.","5B, it can be inferred that FNfn10-3JLCI4 possesses a relatively rapid on-rate for binding to αvβ3.To test the specificity of the interaction between FNfn10-3JCLI4 and αvβ3 integrin, biotinylated FNfn10-3JCLI4 or FNfn10-WT were added to wells coated with equivalent 5 μg/ml concentrations of different purified integrins (FIG.","5D).","In this assay, the detection of plate-bound FNfn10-3JCLI4 was significant only for those wells which had been coated with αvβ3, and not for wells coated with other integrins, including αvβ5, α5β1, α1β1 and αIIbβ3.Example 3 Determining Relative Affinity of FNfn10-3JCLI4 for αvβ3 Integrin In order to examine the relative affinity of FNfn10-3JCLI4, as compared to other αvβ3 binding molecules, a competition ELISA analysis was performed.","In these experiments, biotinylated FNfn10-3JCLI4 at 800 pM was mixed with molar ratios of competitor proteins ranging from 0.01 to 100, and allowed to bind to αvβ3 coated wells (FIGS.","6A and 6B).","Binding of biotinylated FNfn10-3JCLI4 to αvβ3 was unaffected by the presence of the linear integrin-binding peptide (GRGDSPK, SEQ ID No: 27) or FNfn10-WT, even when these molecules were added at 100-fold molar excess (80 nM), relative to FNfn10-3JCLI4.In contrast, unlabelled FNfn10-3JCLI4 and echistatin inhibited binding of biotinylated FNfn10-3JCLI4 at roughly equal equimolar concentrations (IC50 values were 750 and 930 pM, respectively, as calculated by logarithmic curve-fit).","These findings strongly suggest that FNfn10-3JCLI4 and echistatin have roughly equivalent affinities for αvβ3.Echistatin has a published Kd of 330 pM (Kumar et al., 1997).","Example 4 Role of FNfn10-3JCLI4 as a Staining Agent for αvβ3 Integrin Biotinylated FNfn10-3JCLI4 (1 μg/ml) was added to wild-type K562 cells and to K562 cells that had been stably transfected with αvβ3 integrin (generous gift of Dr. S. Blystone).","Surface bound FNfn10-3JCLI4 was then detected using streptavidin-APC and stained cells were enumerated by flow cytometry.","The mean fluorescence intensity of staining on the αvβ3-positive cells was approximately three logs higher than on the non-transfected control cells (FIG.","7A), and concentrations of biotinylated FNfn10-3JCLI4 as low as 0.008 μg/ml were found to be capable of effectively and specifically staining K562-αvβ3 cells.","Having optimized the staining analysis using the K562-αvβ3 cells, a more complete analysis of the specificity of cell surface binding of biotinylated FNfn10-3JCLI4 was performed, using a panel of K562 cell sublines, each of which had been stably transfected with various integrin heterodimers (αvβ3, αvβ5, αIIbβ3, α4β1, and α4β7; all gifts of S. Blystone) (Blystone et al., 1994).","These experiments revealed that FNfn10-3JCLI4 showed a strong selective preference for αvβ3, although it did also react very weakly with αIIbβ3 positive cells.","In contrast, FNfn10-WT showed a slight preference for cells expressing αvβ3, but stained all of the K562 lines with low intensity (FIG.","7B).","One of the unique properties of the FNfn10 scaffold is its high level of physical stability.","Therefore, an examination was performed to determine the effect of prolonged incubation at elevated temperatures on the binding activity of FNfn10-3JCLI4, as compared to the αvβ3s-specific monoclonal antibody, LM609.In these studies, FNfn10-3JCLI4 and LM609 were incubated for 24 hours at 4, 20, 37, 50, 65, or 75 degrees Celsius, prior to addition to K562-αvβ3 cells and performance of flow cytometry.","As can be observed in FIG.","7C, incubation of FNfn10-3JCLI4 at 75° C. for 24 hours had only a modest effect on its cell binding activity; in contrast, the same conditions resulted in the complete elimination of LM609's ability to interact with αvβ3.Example 5 Role of FNfn10-3JCLI4 as an Inhibitor of αvβ3-Dependent Cell Adhesion K562-αvβ3 cells, but not K562 cells, adhere strongly to vitronectin (but not BSA).","This property was therefore used as the basis for a quantitative assessment of the ability of FNfn10-3JLCI4 to disrupt αvβ3 dependent cell adhesion.","To do this, K562-αvβ3 cells were allowed to adhere to vitronectin-coated wells, and the number of bound cells per well was then quantified by crystal violet staining and cell solubilization.","LM609, a commonly used αvβ3 function-blocking antibody, and FNfn10-3JCLI4 were both found to inhibit adhesion of K562-αvβ3 cells to vitronectin, while FNfn10-WT showed little effect in this assay (FIG.","8A).","A more careful analysis, using a wider range of protein concentrations, revealed that FNfn10-3JCLI4 inhibits αvβ3-dependent adhesion to vitronectin at approximately the same concentrations as the well-characterized αvβ3-binding disintegrin, echistatin (FIG.","8B).","Based on an exponential curve-fit, echistatin was found to inhibit αvβ3-mediated cell adhesion to vitronectin with an IC50 of 5.9 nM, while FNfn10-3JCLI4 inhibited αvβ3-dependent adhesion to vitronectin with an IC50 of 8.6 nM (R>0.99 in both cases).","Finally, the effect of FNfn10-3JCLI4 on αIIbβ3-dependent cell adhesion was also examined, because of the weak binding activity of FNfn10-3JCLI4 for K562-αIIbβ3 cells that was revealed in the flow cytometric analyses (Example 4 above).","These experiments revealed that FNfn10-3JCLI4 had no measurable effect on αIIbβ3-mediated cell adhesion to fibronectin, while echistatin inhibited αIIbβ3-dependent adhesion with an IC50 of 10.2 nM (R>0.99; FIG.","8C).","Discussion of Examples 1-5 In this work, phage display technology and random mutagenesis techniques were applied to carry out affinity maturation of FNfn10 binding to αvβ3 integrin.","Using one phage clone that was derived from a biopanning screen against immobilized αvβ3, 3JCLI4, the following was observed: (i) modified FNfn10 phage clones, including 3JCLI4, which bind to αvβ3 integrin all contain an XRGDWXEX (SEQ ID No: 26) consensus and exhibit calcium-dependent binding; (ii) purified FNfn10-3JCLI4 protein binds to immobilized human αvβ3 integrin with much higher affinity than FNfn10-WT and exhibits only background binding to other purified integrins; (iii) binding of FNfn10-3JLCI4 to αvβ3 could be successfully competed using approximately equimolar concentrations of unlabelled FNfn10-3JCLI4 or echistatin, but not by either FNfn10-WT or GRGDSPK peptide (SEQ ID No: 27), even when added at 100-fold molar excess, (iv) FNfn10-3JCLI4 is a sensitive and specific reagent for the detection of human αvβ3 integrin in flow cytometry applications; (v) the binding of FNfn10-3JCLI4 to αvβ3 integrin is maintained even after prolonged exposure of FNfn10-3JCLI4 to high temperature, and (vi) FNfn10-3JCLI4 inhibits cell adhesion via αvβ3 but not αIIbβ3 integrin at nanomolar concentrations.","The RGDW sequence that was detected in the αvβ3-selected FNfn10-display phage clones has previously been reported to be preferentially selective for αIIbβ3 integrin over αvβ3, at least in the context of disintegrins such as eristostatin (McLane et al., 1996).","RGDW peptides are also used to induce integrin activation and in competition assays with putative αIIbβ3 integrin ligands (Hantgan et al., 1992).","Therefore, it was quite unexpected that the consensus sequence (XRGDWXEX, SEQ ID No: 26) bound preferentially to αvβ3 over αIIbβ3 integrins.","Because FNfn10-3JCLI4 exhibits no binding to plate-immobilized αIIbβ3 and very low cross-reactivity with αIIbβ3-positive cells in flow cytometric analysis, the tertiary structure of FNfn10 may play a role in the increased specificity of this RGDW-containing molecule for αvβ3 integrin.","Alternatively, the glutamic acid residue within the extended consensus motif (SEQ ID No: 26) may be significant.","The disintegrins eristostatin and EC6B each contain an aspartic acid residue at the corresponding location (RGDWND, SEQ ID No: 28) (McLane et al., 1996; Scarborough et al., 1993; Marcinkiewicz et al., 2000), and though they both bind αvβ3, they interact preferentially with αIIbβ3.Like the disintegrins, FNfn10-3JCLI4 is a small monomeric molecule with high integrin-binding affinity.","In the above Examples, echistatin and FNfn10-3JCLI4 were inferred to have similar binding affinities for αvβ3 because they were both able to compete with biotinylated FNfn10-3JCLI4 for binding to immobilized αvβ3, at roughly equimolar concentrations.","Consistent with this, echistatin has a reported Kd of 330 pM for αvβ3 (Kumar et al., 1997), while the value calculated here for the half-maximal binding affinity of FNfn10-3JCLI4 for αvβ3 was 800 pM.","Biochemical analyses revealed a rapid rate of association rate between FNfn10-3JCLI4 and αvβ3 (minutes), and a very slow rate of dissociation (>24 hours).","These properties, combined with its high affinity for αvβ3, suggested that FNfn10-3JCLI4 might be suitable for use in flow cytometric applications.","Experimental analysis revealed this was indeed the case, and these studies also showed that FNfn10-3JCLI4 performed similarly to αvβ3-specific monoclonal antibodies in flow cytometric staining using K562-αvβ3 cells.","It should be noted that FNfn10-3JCLI4 is monomeric, unlike monoclonal antibodies (divalent) or other cell-staining reagents used in flow cytometry (MHC:peptide tetramers).","Furthermore, FNfn10-3JCLI4 is considerably more stable than a typical monoclonal antibody, as revealed by its ability to continue to bind αvβ3, even after 24 hr.","incubation at 75° C. A problem encountered in many molecules that target αvβ3 integrins is their cross-reactivity with other integrin subtypes, particularly other αv or β3 integrins.","FNfn10-3JCLI4 was found to bind only to immobilized purified αvβ3, and not to other integrins tested.","The failure of FNfn10-WT and FNfn10-3JCLI4 to interact with α5β1 is consistent with previous reports that the ninth and eighth fibronectin type III domains are also required for FNfn10 binding to α5β1 (Altroff et al., 2001).","In addition, the inability of FNfn10-3JCLI4 to bind to αvβ5 and αIIbβ3, suggests that its integrin-binding activity is likely dependent on a structural interaction with the αvβ3 complex and not with either subunit alone.","The narrow integrin-binding specificity of FNfn10-3JCLI4 can be contrasted with the more broad integrin-binding profile that is exhibited by disintegrins such as eristostatin and kistrin, which bind to both αIIbβ3 and αvβ3 (McLane et al., 1996).","Similarly, the penton base protein from adenovirus type 5 binds to both αvβ3 and αvβ5, and even an engineered Fab construct (WOW-1) has been shown to exhibit cross-reactivity with αvβ5 integrin (Pampori et al., 1999).","Although FNfn10-3JCLI4 did exhibit a weak interaction with αIIbβ3 in the flow cytometry assay, this will not be a major detriment to the more widespread use of this molecule in cell staining applications because (i) no interaction with αIIbβ3 was detected in biochemical binding studies, (ii) binding to αIIbβ3 positive cells was almost 10-fold weaker than binding to αvβ3-positive cells in the flow cytometric analysis, and (iii) αIIbβ3 expression is limited to platelets.","The ability of FNfn10-3JCLI4 to block αvβ3-dependent cell adhesion but not αIIbβ3-mediated cell binding further supports its specificity for αvβ3.The structure of FNfn10-3JCLI4 is advantageous for multiple applications; FNfn10-3JCLI4 lacks disulfide bonds rendering it resistant against reducing agents, is very stable even at high temperature, and can be produced in a bacterial expression system with yields of up to 50-100 mg per liter of culture.","As an endogenous human protein, wild-type fibronectin does not elicit an immune response and it is anticipated that FNfn10-3JCLI4 is likely to be similarly non-immunogenic.","Furthermore, because FNfn10-3JCLI4 is a monomer that lacks disulfide bonds, it has several advantages over a monoclonal antibody with similar affinity.","As a small, single-chain molecule, FNfn10-3JCLI4 could potentially be incorporated into gene delivery vectors (e.g.","viral vectors, liposomes) for cell or tissue-specific gene expression.","Indeed, the incorporation of low-affinity RGD peptides into various viral vectors, and the addition of bispecific antibody complexes to viral surface components, have already been shown to result in desirable improvements in vector specificity and transduction efficiency (Haisma et al., 2000; Wickham et al., 1996; Wickham et al., 1997; Hart, 1999).","αvβ3-positive cells that may be of particular interest for targeted vector systems include dendritic cells (for vaccine applications) as well as osteoclasts and angiogenic vessels.","Finally, FNfn10-3JCLI4 will likely have utility as a cancer imaging agent, because of its ability to recognize αvβ3 on tumor vasculature.","Example 6 Formation of αvβ3-Directed MRI Diagnostic With Superparamagnetic Iron Oxide Particles Polypeptide monobodies prepared as described in Example 1 will be used to form an MRI diagnostic agent upon conjugation to superparamagnetic iron oxide (SPIO) particles.","Dextran-coated SPIO particles can be purchased (Advanced Magnetics, Cambridge, Mass.)","or synthesized as described previously (Kresse et al., 1998).","SPIO particles are extremely effective relaxation agents, especially as effectors of T2*.","The half-life of SPIO particles in blood highly depends on particle size (Moore et al., 2000) and will be an important factor in the design.","A long half-life is useful for allowing time for the contrast agent to escape the capillaries and penetrate into tissue.","However, αvβ3 is highly accessible to intravascular compounds, and a long blood half-life increases a background, non-specific signal enhancement.","Therefore, SPIO particles with diameter >30 nm that have short blood half-lives of 8-20 minutes (Reimer et al., 1992; Moore et al., 2000) will be used.","This should allow sufficient time for binding to αvβ3 and an acceptably short time for clearance of unbound agent.","Particle size is primarily determined by the amount of dextran used during oxide crystal precipitation.","Size distribution will be determined by a dynamic light scattering instrumentation according to known procedures.","Conjugation between the polypeptide monobody and SPIO will be achieved using a streptavidin-biotin linkage.","Two approaches will be employed.","In a first approach, a fusion protein will be expressed from a chimeric gene which includes a polypeptide monobody coding region and streptavidin-biotin coding region linked by an in-frame gene fusion.","The chimeric gene will be inserted into an E. coli expression vector (pET derivative) to produce a biotinylated protein.","In a second approach, a polypeptide monobody will be expressed containing a free cysteine in the C-terminal flexible tail.","Thereafter, the polypeptide monobody will be biotinylated by chemical modification.","Unlike antibodies and cyclic peptides, FN3 have no cysteine residues that would complicate cysteine modification.","These proteins also contain a poly-histidine tag in the C-terminal tail for purification.","The C-terminus of FN3 is located at the opposite side of the protein from the RGD sequence of the FG loop (FIG.","1) and, thus, it is expected that these modification will have little, if any, effect on the binding activity of FN3 mutants.","Streptavidin will be cross-linked with the dextran coat of the SPIO particles by chemical modification (reductive amination) (Shen et al., 1996).","Then, biotinylated FN3 will be attached to this complex, and FN3-biotin-streptavidin-SPIO will be purified using ultrafiltration.","REFERENCES Each of the references cited herein or otherwise listed below are expressly incorporated by reference in their entirety into this specification.","Adonis et al.","(1994) Amer.","J. 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Biol.","Chem.","263, 14621-14624.Zhou et al.","(1991) Biochim.","Biophys.","Acta 1065, 8-14.U.S.","Pat.","No.","6,057,155 to Wickham et al.","U.S. Pat.","No.","6,033,908 to Bout et al.","U.S. Pat.","No.","6,001,557 to Wilson et al.","U.S. Pat.","No.","5,994,132 to Chamberlain et al.","U.S. Pat.","No.","5,981,225 to Kochanek et al.","U.S. Pat.","No.","5,885,808 to Spooner et al.","U.S. Pat.","No.","5,885,613 to Holland et al.","U.S. Pat.","No.","5,871,727 to Curiel U.S. Pat.","No.","5,849,586 to Kriegler et al.","U.S. Pat.","No.","5,681,811 to Ekwuribe U.S. Pat.","No.","5,653,996 to Hsu et al.","U.S. Pat.","No.","5,643,599 to Lee et al.","U.S. Pat.","No.","5,631,237 to Dzau et al.","U.S. Pat.","No.","5,059,421 to Loughrey et al.","U.S. Pat.","No.","4,237,224 to Cohen and Boyer Although the invention has been described in detail for the purposes of illustration, it is understood that such detail is solely for that purpose, and variations can be made therein by those skilled in the art without departing from the spirit and scope of the invention which is defined by the following claims."]],"string":"[\n [\n \"Alphanubeta3 integrin-binding polypeptide monobodies and their use\",\n \"The present invention relates to a polypeptide monobody which includes a modified acid sequence and renders the polypeptide monobody able to bind selectively to αvβ3 integrin.\",\n \"Fusion proteins and conjugates which include the polypeptide monobody, as well as compositions containing the polypeptide monobody, fusion proteins, or conjugates are also disclosed.\",\n \"Uses thereof include: treating or preventing an αvβ3 integrin-mediated disease or disorder, inhibiting αvβ3 integrin activity, treating a cancerous or precancerous condition, imaging tissues using positron emission tomography or magnetic resonance imaging, assessing the metastatic characteristics of a tumor, and delivering DNA to a cell.\"\n ],\n [\n \"1.A polypeptide monobody comprising: at least two Fn3 β-strand domain sequences with a loop region sequence linked between adjacent β-strand domain sequences; and optionally, an N-terminal tail of at least about 2 amino acids, a C-terminal tail of at least about 2 amino acids, or both; wherein at least one loop region sequence, the N-terminal tail, or the C-terminal tail comprises a modified amino acid sequence which varies by deletion, insertion, or replacement of at least two amino acids from a corresponding loop region, N-terminal tail, or C-terminal tail in a wild-type Fn3 domain of fibronectin, and wherein the polypeptide monobody binds selectively to αvβ3 integrin.\",\n \"2.The polypeptide monobody according to claim 1, wherein the modified amino acid sequence comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).\",\n \"3.The polypeptide monobody according to claim 2, wherein the modified amino acid sequence comprises LRGDWSED (SEQ ID No: 5), VRGDWYEY (SEQ ID No: 6), GRGDWTEH (SEQ ID No: 10), ARGDWVEG (SEQ ID No: 11), PRGDWTEG (SEQ ID No: 12), PRGDWIEF (SEQ ID No: 15), PRGDWNEG (SEQ ID No: 23), or FRGDWIEL (SEQ ID No: 24).\",\n \"4.The polypeptide monobody according to claim 1, wherein said at least one loop region sequence is selected from the group consisting of a BC loop region sequence, a DE loop region sequence, an FG loop region sequence, and combinations thereof.\",\n \"5.The polypeptide monobody according to claim 4, wherein said at least one loop region sequence is an FG loop region sequence.\",\n \"6.The polypeptide monobody according to claim 5, wherein the FG loop region sequence comprises, as the modified amino acid sequence, XRGDWXEX where X is any amino acid (SEQ ID No: 26).\",\n \"7.The polypeptide monobody according to claim 6, wherein the modified amino acid sequence comprises LRGDWSED (SEQ ID No: 5), VRGDWYEY (SEQ ID No: 6), GRGDWTEH (SEQ ID No: 10), ARGDWVEG (SEQ ID No: 11), PRGDWTEG (SEQ ID No: 12), PRGDWIEF (SEQ ID No: 15), PRGDWNEG (SEQ ID No: 23), or FRGDWIEL (SEQ ID No: 24).\",\n \"8.A nucleic acid molecule encoding the polypeptide monobody of claim 1.9.The nucleic acid molecule according to claim 8, wherein the nucleic acid is DNA.\",\n \"10.A DNA construct comprising: the DNA molecule of claim 9; a promoter-effective DNA molecule operably coupled 5′ of the DNA molecule; and a transcription termination DNA molecule operably coupled 3′ of the DNA molecule.\",\n \"11.An expression vector into which is inserted a DNA construct according to claim 9.12.A host cell transformed with a DNA construct according to claim 10.13.The host cell according to claim 12, wherein the host cell is a prokaryote.\",\n \"14.The host cell according to claim 12, wherein the host cell is a eukaryote.\",\n \"15.A fusion protein comprising: a polypeptide monobody according to claim 1 and a second polypeptide linked by peptide bond to the polypeptide monobody, the second polypeptide being (i) an epitope tag polypeptide, (ii) a detectable marker polypeptide, (iii) a metal ion-complexing polypeptide, or (iv) a DNA-binding polypeptide.\",\n \"16.The fusion protein according to claim 15 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX (SEQ ID No: 26) where X is any amino acid.\",\n \"17.The fusion protein according to claim 15 wherein the second polypeptide is an epitope tag polypeptide.\",\n \"18.The fusion protein according to claim 17 wherein the epitope tag comprises a polyhistidine amino acid sequence.\",\n \"19.The fusion protein according to claim 15 wherein the second polypeptide is a detectable marker polypeptide.\",\n \"20.The fusion protein according to claim 19 wherein the detectable marker polypeptide is an alkaline phosphatase.\",\n \"21.The fusion protein according to claim 15 wherein the second polypeptide is a metal ion-complexing polypeptide.\",\n \"22.The fusion protein according to claim 21 wherein the metal-ion complexing polypeptide is a polyhistidine amino acid sequence.\",\n \"23.The fusion protein according to claim 15 wherein the second polypeptide is a DNA-binding polypeptide.\",\n \"24.The fusion protein according to claim 23 wherein the DNA-binding polypeptide comprises a polylysine amino acid sequence.\",\n \"25.A fusion protein-metal ion complex comprising a fusion protein according to claim 21 which is complexed with a metal ion.\",\n \"26.The fusion protein-metal ion complex according to claim 25 wherein the metal ion is a positron emitting metal ion, a paramagnetic metal ion, a radioactive metal ion, or a non-radioactive metal ion.\",\n \"27.The fusion protein-metal ion complex according to claim 25 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).\",\n \"28.A fusion protein-DNA complex comprising a fusion protein according to claim 23 which is complexed with a DNA molecule via the DNA-binding polypeptide.\",\n \"29.The fusion protein-DNA complex according to claim 28 wherein the DNA-binding polypeptide comprises a polylysine amino acid sequence.\",\n \"30.The fusion protein-DNA complex according to claim 28 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).\",\n \"31.A nucleic acid molecule encoding the fusion protein of claim 15.32.The nucleic acid molecule according to claim 31, wherein the nucleic acid is DNA.\",\n \"33.A DNA construct comprising: the DNA molecule of claim 32; a promoter-effective DNA molecule operably coupled 5′ of the DNA molecule; and a transcription termination DNA molecule operably coupled 3′ of the DNA molecule.\",\n \"34.An expression vector into which is inserted a DNA construct according to claim 33.35.A host cell transformed with a DNA construct according to claim 33.36.The host cell according to claim 35, wherein the host cell is a prokaryote.\",\n \"37.The host cell according to claim 35, wherein the host cell is a eukaryote.\",\n \"38.A conjugate comprising a polypeptide monobody according to claim 1 conjugated to (i) a chemotherapeutic agent, (ii) a contrasting agent, or (iii) an organic chelating agent.\",\n \"39.The conjugate according to claim 38 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).\",\n \"40.The conjugate according to claim 38 wherein the polypeptide monobody is conjugated to a chemotherapeutic agent.\",\n \"41.The conjugate according to claim 38 wherein the polypeptide monobody is conjugated to a contrasting agent or an organic chelating agent.\",\n \"42.The conjugate according to claim 41, wherein the contrasting agent is a dextran-coated superparamagnetic iron oxide particle, a lipid vesicle with Gd3+ attached to a surface thereof, or poly-lysine with Gd3+ attached to an amino group thereof 43.The conjugate according to claim 41, wherein the organic chelating agent is diethylenetriamine pentaacetic acid.\",\n \"44.A composition comprising a pharmaceutically acceptable carrier and a polypeptide monobody according to claim 1.45.A composition comprising a pharmaceutically acceptable carrier and the fusion protein according to claim 15.46.A composition comprising a pharmaceutically acceptable carrier and the conjugate according to claim 38.47.A method of treating or preventing an αvβ3 integrin-mediated disease or disorder comprising: administering to a patient in need thereof, an effective amount of a polypeptide or polypeptide monobody according to claim 1 which binds to the αvβ3 integrin to inhibit activity thereof, thereby treating or preventing the αvβ3 integrin-mediated disease or disorder.\",\n \"48.The method according to claim 47 wherein said administering is carried out under conditions which are effective to contact one or more cells expressing αvβ3 integrin with a sufficient amount of the polypeptide monobody to inhibit the activity of αvβ3 integrin on such one or more cells.\",\n \"49.The method according to claim 47 wherein the αvβ3 integrin-mediated disease or disorder is tumor metastasis, solid tumor growth, osteoporosis, Paget's disease, humoral hypercalcemia of malignancy, angiogenesis, retinopathy, arthritis, periodontal disease, psoriasis, smooth muscle cell migration, viral infection, fungal infection, and bacterial infection.\",\n \"50.The method according to claim 47 wherein the αvβ3 integrin-mediated disease or disorder is tumor metastasis or solid tumor growth, the polypeptide monobody being present in the form of a conjugate comprising the polypeptide monobody conjugated to a chemotherapeutic agent.\",\n \"51.The method according to claim 47 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).\",\n \"52.A method of treating a cancerous or precancerous condition comprising: administering to a patient in need thereof an effective amount of a polypeptide monobody according to claim 1, wherein the polypeptide monobody binds to cancerous or precancerous cells expressing αvβ3 integrin to inhibit αvβ3 integrin-induced activity of the cancerous or precancerous cells.\",\n \"53.The method according to claim 52 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).\",\n \"54.A method of treating a cancerous or precancerous condition comprising: administering to a patient in need thereof an effective amount of a conjugate according to claim 40, wherein the polypeptide monobody binds to cancerous or precancerous cells expressing αvβ3 integrin to inhibit αvβ3 integrin-induced activity of the cancerous or precancerous cells and delivers the chemotherapeutic agent to the cancerous or precancerous cells.\",\n \"55.The method according to claim 54 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).\",\n \"56.A method of imaging tissues using positron emission tomography or magnetic resonance imaging, the method comprising: administering a fusion protein-metal ion complex according to claim 25 to a patient and detecting, by positron emission tomography or magnetic resonance imaging, whether the conjugate is localized within any tissues of the patient.\",\n \"57.The method according to claim 56 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).\",\n \"58.A method of imaging tissues using positron emission tomography or magnetic resonance imaging, the method comprising: administering a conjugate according to claim 41 to a patient and detecting, by positron emission tomography or magnetic resonance imaging, whether the conjugate is localized within any tissues of the patient.\",\n \"59.The method according to claim 58 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).\",\n \"60.A method of assessing the metastatic characteristics of a tumor, the method comprising: administering a fusion protein-metal ion complex according to claim 25 to a patient and detecting, by positron emission tomography or magnetic resonance imaging, whether the conjugate is localized within any tissues of the patient, wherein localization of the fusion protein-metal ion complex indicates an angiogenic site containing a potentially metastatic tumor.\",\n \"61.The method according to claim 60 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).\",\n \"62.A method of assessing the metastatic characteristics of a tumor, the method comprising: administering a conjugate according to claim 41 to a patient and detecting, by positron emission tomography or magnetic resonance imaging, whether the conjugate is localized within any tissues of the patient, wherein localization of the conjugate indicates an angiogenic site containing a potentially metastatic tumor.\",\n \"63.The method according to claim 62 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).\",\n \"64.A method of inhibiting αvβ3 integrin activity comprising: contacting αvβ3 integrin with a polypeptide monobody according to claim 1 under conditions effective to inhibit the activity of the αvβ3 integrin.\",\n \"65.The method according to claim 64 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).\",\n \"66.A method of delivering DNA to a cell comprising: providing a fusion protein-DNA complex according to claim 28 and contacting a cell expressing an αvβ3 integrin with the fusion protein-DNA complex under conditions effective to cause uptake of the fusion protein-DNA complex, thereby delivering the DNA into the cell.\",\n \"67.The method according to claim 66 wherein the modified amino acid sequence of the polypeptide monobody comprises XRGDWXEX where X is any amino acid (SEQ ID No: 26).\",\n \"68.An isolated polypeptide which binds to an αvβ3 integrin and comprises the amino acid sequence XRGDWXEX where X is any amino acid (SEQ ID No: 26).\",\n \"69.The polypeptide according to claim 68, wherein the polypeptide comprises the amino acid sequence of LRGDWSED (SEQ ID No: 5), VRGDWYEY (SEQ ID No: 6), GRGDWTEH (SEQ ID No: 10), ARGDWVEG (SEQ ID No: 11), PRGDWTEG (SEQ ID No: 12), PRGDWIEF (SEQ ID No: 15), PRGDWNEG (SEQ ID No: 23), or FRGDWIEL (SEQ ID No: 24).\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>αvβ3 (CD51/CD61) is a member of the integrin family of cell surface adhesion receptors.\",\n \"Over 20 different αβ integrin heterodimers exist, each with different tissue and ligand specificities.\",\n \"Normal tissue distribution of αvβ3 is generally limited to high levels of expression on osteoclasts, with lower levels observed on platelets, megakaryocytes, kidney, vascular smooth muscle, placenta, dendritic cells (Weiss et al., 2001), and in varying amounts on normal endothelium (reviewed in Horton, 1997).\",\n \"In contrast to α5β1, which binds to only fibronectin, αvβ3 binds to a wide range of RGD-containing integrin ligands, including but not limited to fibronectin, vitronectin, osteopontin, von Willebrandt factor, and fibrinogen (Horton, 1997).\",\n \"αvβ3 integrin is a multifunctional cell surface receptor that has pleiotropic roles in normal cell growth and survival.\",\n \"αvβ3 integrin can also contribute to oncogenesis.\",\n \"Consistent with this, upregulation of αvβ3 expression has been observed on the endothelial cells of angiogenic vessels, and binding of αvβ3 to the basement membrane is a critical step in the angiogenesis induced by basic fibroblast growth factor and tumor necrosis factor-α (Friedlander et al., 1995).\",\n \"Expression of αvβ3 has also been implicated in tumor invasion, and it has been shown that αvβ3 binds matrix metalloproteinase-2 (MM-2) and presents MMP-2 on the surface of invasive carcinomas and on invasive angiogenic endothelial cells (Brooks et al., 1996; Silletti et al., 2001).\",\n \"αvβ3 also regulates cell growth and survival, since ligation of this receptor can, under some circumstances, induce apoptosis in tumor cells (Kozlova et al., 2001).\",\n \"Furthermore, disruption of cell adhesion with anti-αvβ3 antibodies, RGD peptides, and other integrin antagonists has been shown to slow tumor growth (Chatterjee et al., 2000; Chatterjee et al., 2001; and Brooks et al., 1996).\",\n \"Finally, the selective upregulation of αvβ3 expression on tumor blood vessels is also being explored as the basis for imaging of neoplastic lesion, and the αvβ3-specific antibody LM609 has been successfully used for this purpose in vivo (Sipkins et al., 1998).\",\n \"Novel molecules capable of binding with high specificity to αvβ3 integrin have potential utility in several applications, and as a consequence, αvβ3 has been a frequent target for drug discovery and selection of new binding ligands.\",\n \"Phage display technology, in which combinatorial peptide libraries are expressed on the surface of bacteriophages, has been used to select for peptide ligands capable of binding to αvβ3—yielding a wide range of RGD-containing peptide sequences capable of interacting at moderate affinity with αvβ3 and other integrins (Koivunen et al., 1994; Healy et al., 1995).\",\n \"Phage-displayed random peptide libraries have also been constructed and screened using framework proteins such as the cytotoxic T lymphocyte-associated antigen 4 (CTLA-4).\",\n \"This resulted in the identification of phage clones which could be used to stain human umbilical vein endothelial cells in a flow cytometric assay.\",\n \"However, the ability of the purified recombinant CTLA-4 protein to stain cells, and its cross-reactivity with other integrins was not reported (Hufton et al., 2000).\",\n \"Fibronectin is a natural ligand of integrins, and it contains repeats of three types of domains.\",\n \"The tenth fibronectin type III domain (FNfn10) includes the RGD sequence in the loop connecting the F and G β-strands (FG loop) (Main et al., 1992).\",\n \"FNfn10 was developed as a scaffold for phage display of peptides because of its small size (94 residues), monomeric assembly, and ability to retain its global fold while exposed loops were randomized (Koide et al., 1998).\",\n \"In addition, FNfn10 lacks cysteine residues and requires no post-translational modification, allowing for large-scale bacterial expression.\",\n \"It has been shown that residues in the FG loop including the RGD sequence are highly flexible (Main et al., 1992; Carr et al., 1997) and this flexibility of the FG loop has been implicated as the origin of the ability of FNfn10 to interact with multiple integrins (Main et al., 1992).\",\n \"While the stability of monobodies makes them well suited for intracellular studies, there has been no prior use of monobodies to probe for modified FG loop or other loop sequences with enhanced binding affinity for and selectivity between integrins, particularly αvβ3 integrin.\",\n \"The present invention overcomes these and other deficiencies in the art.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>A first aspect of the present invention relates to a polypeptide monobody which includes: at least two Fn3 β-strand domain sequences with a loop region sequence linked between adjacent β-strand domain sequences; and optionally, an N-terminal tail of at least about 2 amino acids, a C-terminal tail of at least about 2 amino acids, or both; wherein at least one loop region sequence, the N-terminal tail, or the C-terminal tail comprises a modified amino acid sequence which varies by deletion, insertion, or replacement of at least two amino acids from a corresponding loop region, N-terminal tail, or C-terminal tail in a wild-type Fn3 domain of fibronectin, and wherein the polypeptide monobody binds selectively to αvβ3 integrin.\",\n \"Compositions containing the polypeptide monobody are also disclosed.\",\n \"A second aspect of the present invention relates to a nucleic acid molecule encoding a polypeptide monobody of the present invention.\",\n \"A third aspect of the present invention relates to a DNA construct which includes a DNA molecule encoding a polypeptide monobody of the present invention; a promoter-effective DNA molecule operably coupled 5′ of the DNA molecule; and a transcription termination DNA molecule operably coupled 3′ of the DNA molecule.\",\n \"Expression vectors and host cells which include the DNA construct are also disclosed.\",\n \"A fourth aspect of the present invention relates to a fusion protein which includes: a polypeptide monobody of the present invention and a second polypeptide linked by peptide bond to the polypeptide monobody, the second polypeptide being (i) an epitope tag polypeptide, (ii) a detectable marker polypeptide, (iii) a metal ion-complexing polypeptide, or (iv) a DNA-binding polypeptide.\",\n \"Compositions containing the fusion protein are also disclosed.\",\n \"A fifth aspect of the present invention relates to a fusion protein-metal ion complex which includes a fusion protein of the present invention including a metal ion-complexing polypeptide as the second polypeptide, where the metal ion-complexing polypeptide is complexed with a metal ion.\",\n \"A sixth aspect of the present invention relates to a fusion protein-DNA complex which includes a fusion protein of the present invention including a DNA-binding polypeptide as the second polypeptide, where the DNA-binding polypeptide is complexed with a DNA molecule via the DNA-binding polypeptide.\",\n \"A seventh aspect of the present invention relates to a nucleic acid molecule encoding a fusion protein of the present invention.\",\n \"An eighth aspect of the present invention relates to a DNA construct which includes a DNA molecule encoding a fusion protein of the present invention; a promoter-effective DNA molecule operably coupled 5′ of the DNA molecule; and a transcription termination DNA molecule operably coupled 3′ of the DNA molecule.\",\n \"Expression vectors and host cells which include the DNA construct as also disclosed.\",\n \"A ninth aspect of the present invention relates to a conjugate comprising a polypeptide monobody of the present invention conjugated to (i) a chemotherapeutic agent, (ii) a contrasting agent, or (iii) an organic chelating agent.\",\n \"Compositions containing the conjugate are also disclosed.\",\n \"A tenth aspect of the present invention relates to a method of treating or preventing an αvβ3 integrin-mediated disease or disorder which includes: administering to a patient in need thereof an effective amount of a polypeptide monobody of the present invention which binds to the αvβ3 integrin to inhibit activity thereof, thereby treating or preventing the αvβ3 integrin-mediated disease or disorder.\",\n \"An eleventh aspect of the present invention relates to a method of treating a cancerous or precancerous condition which includes: administering to a patient in need thereof an effective amount of a polypeptide monobody of the present invention, wherein the polypeptide monobody binds to cancerous or precancerous cells expressing αvβ3 integrin to inhibit αvβ3 integrin-induced activity of the cancerous or precancerous cells.\",\n \"A twelfth aspect of the present invention relates to a method of treating a cancerous or precancerous condition which includes: administering to a patient in need thereof an effective amount of a conjugate of the present invention, wherein the polypeptide monobody binds to cancerous or precancerous cells expressing αvβ3 integrin to inhibit αvβ3 integrin-induced activity of the cancerous or precancerous cells and delivers the chemotherapeutic agent to the cancerous or precancerous cells.\",\n \"A thirteenth aspect of the present invention relates to a method of imaging tissues using positron emission tomography or magnetic resonance imaging, the method including: administering a fusion protein-metal ion complex of the present invention to a patient and detecting, by positron emission tomography or magnetic resonance imaging, whether the complex is localized within any tissues of the patient.\",\n \"A fourteenth aspect of the present invention relates to a method of imaging tissues using positron emission tomography or magnetic resonance imaging, the method including: administering to a patient a conjugate of the present invention which includes a contrasting agent and detecting, by positron emission tomography or magnetic resonance imaging, whether the conjugate is localized within any tissues of the patient.\",\n \"A fifteenth aspect of the present invention relates to a method of assessing the metastatic characteristics of a tumor, the method including: administering a fusion protein-metal ion complex of the present invention to a patient and detecting, by positron emission tomography or magnetic resonance imaging, whether the conjugate is localized within any tissues of the patient, wherein localization of the fusion protein-metal ion complex indicates an angiogenic site containing a potentially metastatic tumor.\",\n \"A sixteenth aspect of the present invention relates to a method of assessing the metastatic characteristics of a tumor, the method including: administering to a patient a conjugate of the present invention which includes a contrasting agent and detecting, by positron emission tomography or magnetic resonance imaging, whether the conjugate is localized within any tissues of the patient, wherein localization of the fusion protein-metal ion complex indicates an angiogenic site containing a potentially metastatic tumor.\",\n \"A seventeenth aspect of the present invention relates to a method of inhibiting αvβ3 integrin activity which includes: contacting αvβ3 integrin with a polypeptide monobody of the present invention under conditions effective to inhibit the activity of the αvβ3 integrin.\",\n \"An eighteenth aspect of the present invention relates to a method of delivering DNA to a cell which includes: providing a fusion protein-DNA complex of the present invention and contacting a cell expressing an αvβ3 integrin with the fusion protein-DNA complex under conditions effective to cause uptake of the fusion protein-DNA complex, thereby delivering the DNA into the cell.\",\n \"A nineteenth aspect of the present invention relates to an isolated polypeptide which binds to an αvβ3 integrin and includes the amino acid sequence XRGDWXEX where X is any amino acid (SEQ ID No: 26).\",\n \"The structure of the polypeptide monobodies of the present invention are advantageous for multiple applications because they (1) lack disulfide bonds, rendering them resistant against reducing agents; (2) are very stable even at high temperature; and (3) can be produced in a bacterial expression system with yields of up to 50-100 mg per liter of culture.\",\n \"Moreover, because they are derived from the endogenous human protein fibronectin, which is non-immunogenic, it is believed that the polypeptide monobodies will not elicit an immune response.\",\n \"As a small, single-chain molecule, DNA encoding the polypeptide monobodies can be incorporated into gene delivery vectors (e.g.\",\n \"viral vectors, liposomes) for cell or tissue-specific gene expression.\",\n \"As a result of these properties, the polypeptide monobodies of the present invention can be used to treat or prevent a number of αvβ3 integrin-associated diseases or disorders as well as for diagnostic imaging of αvβ3 integrin-expressing tissues.\"\n ],\n [\n \"This application claims the benefit of U.S.\",\n \"Provisional Patent Application Ser.\",\n \"No.\",\n \"60/281,481, filed Apr.\",\n \"4, 2001, which is hereby incorporated by reference in its entirety.\",\n \"This invention was made, at least in part, with funding received from (1) the National Institutes of Health grant numbers R29-GM55042, T32-AI07362, and T32-GM07356; and (2) the U.S. Army grant numbers DAMD17-99-1-9361, DAMD17-01-1-0384, and DAMD17-01-1-0385.The U.S. government may have certain rights in this invention.\",\n \"FIELD OF THE INVENTION The present invention relates generally to polypeptide monobodies, more particularly polypeptide monobodies derived from the tenth fibronectin type III domain from human fibronectin (“FNfn10”) which have activity in binding to αvβ3 integrin, fusion proteins or conjugates which include such a polypeptide monobody, as well as the uses thereof for modulating αvβ3 integrin-mediated activity.\",\n \"BACKGROUND OF THE INVENTION αvβ3 (CD51/CD61) is a member of the integrin family of cell surface adhesion receptors.\",\n \"Over 20 different αβ integrin heterodimers exist, each with different tissue and ligand specificities.\",\n \"Normal tissue distribution of αvβ3 is generally limited to high levels of expression on osteoclasts, with lower levels observed on platelets, megakaryocytes, kidney, vascular smooth muscle, placenta, dendritic cells (Weiss et al., 2001), and in varying amounts on normal endothelium (reviewed in Horton, 1997).\",\n \"In contrast to α5β1, which binds to only fibronectin, αvβ3 binds to a wide range of RGD-containing integrin ligands, including but not limited to fibronectin, vitronectin, osteopontin, von Willebrandt factor, and fibrinogen (Horton, 1997).\",\n \"αvβ3 integrin is a multifunctional cell surface receptor that has pleiotropic roles in normal cell growth and survival.\",\n \"αvβ3 integrin can also contribute to oncogenesis.\",\n \"Consistent with this, upregulation of αvβ3 expression has been observed on the endothelial cells of angiogenic vessels, and binding of αvβ3 to the basement membrane is a critical step in the angiogenesis induced by basic fibroblast growth factor and tumor necrosis factor-α (Friedlander et al., 1995).\",\n \"Expression of αvβ3 has also been implicated in tumor invasion, and it has been shown that αvβ3 binds matrix metalloproteinase-2 (MM-2) and presents MMP-2 on the surface of invasive carcinomas and on invasive angiogenic endothelial cells (Brooks et al., 1996; Silletti et al., 2001).\",\n \"αvβ3 also regulates cell growth and survival, since ligation of this receptor can, under some circumstances, induce apoptosis in tumor cells (Kozlova et al., 2001).\",\n \"Furthermore, disruption of cell adhesion with anti-αvβ3 antibodies, RGD peptides, and other integrin antagonists has been shown to slow tumor growth (Chatterjee et al., 2000; Chatterjee et al., 2001; and Brooks et al., 1996).\",\n \"Finally, the selective upregulation of αvβ3 expression on tumor blood vessels is also being explored as the basis for imaging of neoplastic lesion, and the αvβ3-specific antibody LM609 has been successfully used for this purpose in vivo (Sipkins et al., 1998).\",\n \"Novel molecules capable of binding with high specificity to αvβ3 integrin have potential utility in several applications, and as a consequence, αvβ3 has been a frequent target for drug discovery and selection of new binding ligands.\",\n \"Phage display technology, in which combinatorial peptide libraries are expressed on the surface of bacteriophages, has been used to select for peptide ligands capable of binding to αvβ3—yielding a wide range of RGD-containing peptide sequences capable of interacting at moderate affinity with αvβ3 and other integrins (Koivunen et al., 1994; Healy et al., 1995).\",\n \"Phage-displayed random peptide libraries have also been constructed and screened using framework proteins such as the cytotoxic T lymphocyte-associated antigen 4 (CTLA-4).\",\n \"This resulted in the identification of phage clones which could be used to stain human umbilical vein endothelial cells in a flow cytometric assay.\",\n \"However, the ability of the purified recombinant CTLA-4 protein to stain cells, and its cross-reactivity with other integrins was not reported (Hufton et al., 2000).\",\n \"Fibronectin is a natural ligand of integrins, and it contains repeats of three types of domains.\",\n \"The tenth fibronectin type III domain (FNfn10) includes the RGD sequence in the loop connecting the F and G β-strands (FG loop) (Main et al., 1992).\",\n \"FNfn10 was developed as a scaffold for phage display of peptides because of its small size (94 residues), monomeric assembly, and ability to retain its global fold while exposed loops were randomized (Koide et al., 1998).\",\n \"In addition, FNfn10 lacks cysteine residues and requires no post-translational modification, allowing for large-scale bacterial expression.\",\n \"It has been shown that residues in the FG loop including the RGD sequence are highly flexible (Main et al., 1992; Carr et al., 1997) and this flexibility of the FG loop has been implicated as the origin of the ability of FNfn10 to interact with multiple integrins (Main et al., 1992).\",\n \"While the stability of monobodies makes them well suited for intracellular studies, there has been no prior use of monobodies to probe for modified FG loop or other loop sequences with enhanced binding affinity for and selectivity between integrins, particularly αvβ3 integrin.\",\n \"The present invention overcomes these and other deficiencies in the art.\",\n \"SUMMARY OF THE INVENTION A first aspect of the present invention relates to a polypeptide monobody which includes: at least two Fn3 β-strand domain sequences with a loop region sequence linked between adjacent β-strand domain sequences; and optionally, an N-terminal tail of at least about 2 amino acids, a C-terminal tail of at least about 2 amino acids, or both; wherein at least one loop region sequence, the N-terminal tail, or the C-terminal tail comprises a modified amino acid sequence which varies by deletion, insertion, or replacement of at least two amino acids from a corresponding loop region, N-terminal tail, or C-terminal tail in a wild-type Fn3 domain of fibronectin, and wherein the polypeptide monobody binds selectively to αvβ3 integrin.\",\n \"Compositions containing the polypeptide monobody are also disclosed.\",\n \"A second aspect of the present invention relates to a nucleic acid molecule encoding a polypeptide monobody of the present invention.\",\n \"A third aspect of the present invention relates to a DNA construct which includes a DNA molecule encoding a polypeptide monobody of the present invention; a promoter-effective DNA molecule operably coupled 5′ of the DNA molecule; and a transcription termination DNA molecule operably coupled 3′ of the DNA molecule.\",\n \"Expression vectors and host cells which include the DNA construct are also disclosed.\",\n \"A fourth aspect of the present invention relates to a fusion protein which includes: a polypeptide monobody of the present invention and a second polypeptide linked by peptide bond to the polypeptide monobody, the second polypeptide being (i) an epitope tag polypeptide, (ii) a detectable marker polypeptide, (iii) a metal ion-complexing polypeptide, or (iv) a DNA-binding polypeptide.\",\n \"Compositions containing the fusion protein are also disclosed.\",\n \"A fifth aspect of the present invention relates to a fusion protein-metal ion complex which includes a fusion protein of the present invention including a metal ion-complexing polypeptide as the second polypeptide, where the metal ion-complexing polypeptide is complexed with a metal ion.\",\n \"A sixth aspect of the present invention relates to a fusion protein-DNA complex which includes a fusion protein of the present invention including a DNA-binding polypeptide as the second polypeptide, where the DNA-binding polypeptide is complexed with a DNA molecule via the DNA-binding polypeptide.\",\n \"A seventh aspect of the present invention relates to a nucleic acid molecule encoding a fusion protein of the present invention.\",\n \"An eighth aspect of the present invention relates to a DNA construct which includes a DNA molecule encoding a fusion protein of the present invention; a promoter-effective DNA molecule operably coupled 5′ of the DNA molecule; and a transcription termination DNA molecule operably coupled 3′ of the DNA molecule.\",\n \"Expression vectors and host cells which include the DNA construct as also disclosed.\",\n \"A ninth aspect of the present invention relates to a conjugate comprising a polypeptide monobody of the present invention conjugated to (i) a chemotherapeutic agent, (ii) a contrasting agent, or (iii) an organic chelating agent.\",\n \"Compositions containing the conjugate are also disclosed.\",\n \"A tenth aspect of the present invention relates to a method of treating or preventing an αvβ3 integrin-mediated disease or disorder which includes: administering to a patient in need thereof an effective amount of a polypeptide monobody of the present invention which binds to the αvβ3 integrin to inhibit activity thereof, thereby treating or preventing the αvβ3 integrin-mediated disease or disorder.\",\n \"An eleventh aspect of the present invention relates to a method of treating a cancerous or precancerous condition which includes: administering to a patient in need thereof an effective amount of a polypeptide monobody of the present invention, wherein the polypeptide monobody binds to cancerous or precancerous cells expressing αvβ3 integrin to inhibit αvβ3 integrin-induced activity of the cancerous or precancerous cells.\",\n \"A twelfth aspect of the present invention relates to a method of treating a cancerous or precancerous condition which includes: administering to a patient in need thereof an effective amount of a conjugate of the present invention, wherein the polypeptide monobody binds to cancerous or precancerous cells expressing αvβ3 integrin to inhibit αvβ3 integrin-induced activity of the cancerous or precancerous cells and delivers the chemotherapeutic agent to the cancerous or precancerous cells.\",\n \"A thirteenth aspect of the present invention relates to a method of imaging tissues using positron emission tomography or magnetic resonance imaging, the method including: administering a fusion protein-metal ion complex of the present invention to a patient and detecting, by positron emission tomography or magnetic resonance imaging, whether the complex is localized within any tissues of the patient.\",\n \"A fourteenth aspect of the present invention relates to a method of imaging tissues using positron emission tomography or magnetic resonance imaging, the method including: administering to a patient a conjugate of the present invention which includes a contrasting agent and detecting, by positron emission tomography or magnetic resonance imaging, whether the conjugate is localized within any tissues of the patient.\",\n \"A fifteenth aspect of the present invention relates to a method of assessing the metastatic characteristics of a tumor, the method including: administering a fusion protein-metal ion complex of the present invention to a patient and detecting, by positron emission tomography or magnetic resonance imaging, whether the conjugate is localized within any tissues of the patient, wherein localization of the fusion protein-metal ion complex indicates an angiogenic site containing a potentially metastatic tumor.\",\n \"A sixteenth aspect of the present invention relates to a method of assessing the metastatic characteristics of a tumor, the method including: administering to a patient a conjugate of the present invention which includes a contrasting agent and detecting, by positron emission tomography or magnetic resonance imaging, whether the conjugate is localized within any tissues of the patient, wherein localization of the fusion protein-metal ion complex indicates an angiogenic site containing a potentially metastatic tumor.\",\n \"A seventeenth aspect of the present invention relates to a method of inhibiting αvβ3 integrin activity which includes: contacting αvβ3 integrin with a polypeptide monobody of the present invention under conditions effective to inhibit the activity of the αvβ3 integrin.\",\n \"An eighteenth aspect of the present invention relates to a method of delivering DNA to a cell which includes: providing a fusion protein-DNA complex of the present invention and contacting a cell expressing an αvβ3 integrin with the fusion protein-DNA complex under conditions effective to cause uptake of the fusion protein-DNA complex, thereby delivering the DNA into the cell.\",\n \"A nineteenth aspect of the present invention relates to an isolated polypeptide which binds to an αvβ3 integrin and includes the amino acid sequence XRGDWXEX where X is any amino acid (SEQ ID No: 26).\",\n \"The structure of the polypeptide monobodies of the present invention are advantageous for multiple applications because they (1) lack disulfide bonds, rendering them resistant against reducing agents; (2) are very stable even at high temperature; and (3) can be produced in a bacterial expression system with yields of up to 50-100 mg per liter of culture.\",\n \"Moreover, because they are derived from the endogenous human protein fibronectin, which is non-immunogenic, it is believed that the polypeptide monobodies will not elicit an immune response.\",\n \"As a small, single-chain molecule, DNA encoding the polypeptide monobodies can be incorporated into gene delivery vectors (e.g.\",\n \"viral vectors, liposomes) for cell or tissue-specific gene expression.\",\n \"As a result of these properties, the polypeptide monobodies of the present invention can be used to treat or prevent a number of αvβ3 integrin-associated diseases or disorders as well as for diagnostic imaging of αvβ3 integrin-expressing tissues.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS FIG.\",\n \"1 is a schematic drawing of FNfn10 (Main et al., 1992) showing the RGD sequence and the positions of residues in the FG loop that were diversified in the JCFNRGD library.\",\n \"This figure was made with the program MOLSCRIPT (Kraulis, 1991).\",\n \"FIG.\",\n \"2 illustrates a nucleotide sequence (SEQ ID No: 1) encoding the amino acid sequence (SEQ ID No: 2) of the wild-type FNfn10.The amino acid numbering is according to Main et al.\",\n \"(1992).\",\n \"The BC loop region and the FG loop region are shown in boxes.\",\n \"FIGS.\",\n \"3A-B illustrate the amino acid sequence of the wild-type FNfn10 (SEQ ID No: 2, FIG.\",\n \"3A) as well as a mutant FNfn10 (SEQ ID No: 3, FIG.\",\n \"3B) which has the Asp-7 residue replaced with a non-negatively charged amino acid residue (X), which is preferably either Asn or Lys.\",\n \"As reported in Koide et al.\",\n \"(2001), both of these mutations have the effect of promoting greater stability of the mutant FNfn10 at neutral pH as compared to the wild-type FNfn10.FIGS.\",\n \"4A-B illustrate the FG loop sequences used during phage display screening of a JCFN-RGD library.\",\n \"M13 Phage displaying a library of FNfn10 clones with partially randomized FG loops (XRGDXXXX, SEQ ID No: 4) were amplified using 0.0, 0.1, or 1 mM IPTG to vary FNfn10 copy number on the phage.\",\n \"Amplified phage populations were subjected to three rounds of biopanning against wells coated with αvβ3 integrin (5 μg/ml).\",\n \"FIG.\",\n \"4A shows the FG loop sequences for five clones, each of which was sequenced from the second round eluate of the library amplified with 1 mM IPTG (2JCAV1-2JCAV5, SEQ ID Nos: 5-9), from the third round eluate of the library amplified with 1 mM IPTG (3JCAV1-5, SEQ ID Nos: 10-14), from the third round eluate of the library amplified with low IPTG (3JCLI1-5, SEQ ID Nos: 15-19), and from the third round eluate of the library amplified with no IPTG (3JCNI1-5, SEQ ID Nos: 20-24).\",\n \"FIG.\",\n \"4B illustrates the phage binding assay of the phage clones containing the XRGDWXEX consensus (SEQ ID No: 26) and unmodified FNfn10 (pVIII-FNfn10), all amplified without IPTG.\",\n \"Phage inputs of 109, 108, or 107 pfu were added to wells coated with 5 μg/ml αvβ3 (solid bars) or 0.5 mg/ml BSA (patterned bars).\",\n \"After one hour, nonbinding phage were removed by extensive rinsing and bound phage were eluted with 0.2M glycine, pH 2.2.Eluates from each well were titered.\",\n \"Results shown are representative of three different experiments that yielded similar results.\",\n \"FIGS.\",\n \"5A-D are graphs illustrating the assessment of FNfn10-3JCLI4 binding affinity to αvβ3 integrin by ELISA.\",\n \"To determine the half-maximal binding concentration of biotinylated FNfn10-3JCLI4 versus biotinylated FNfn10-WT, wells of a 96-well plate were coated αvβ3 (5 μg/ml), α1β1 (5 μg/ml), or BSA (0.5 mg/ml) and blocked with BSA (FIG.\",\n \"5A).\",\n \"Biotinylated FNfn10-3JCLI4 and FNfn10-WT were then added to the wells and allowed to bind for two hours.\",\n \"Bound protein was detected by streptavidin-horseradish peroxidase (2 μg/ml) and ABTS substrate.\",\n \"O.D.400 measurements were recorded for biotinylated FNfn10-3JCLI4 binding to αvβ3 (⋄), to α1β1 (♦), or to BSA (×) and biotinylated FNfn10-WT binding to αvβ3 (□).\",\n \"Error bars reflect the standard deviation between triplicate wells.\",\n \"To examine protein dissociation over a prolonged (24 hour) time period, a 96-well plate was coated with 5 μg/ml αvβ3 integrin and then incubated with 0.02 μg/ml biotinylated FNfn10-3JCLI4 for two hours; as a control, 10 mM EDTA was added to certain wells, in order to calculate the background level of FNfn10-3JCLI4 binding (FIG.\",\n \"5B).\",\n \"After rinsing, 2 μg/ml unlabelled FNfn10-3JCLI4 was added to the preformed protein complexes present in the wells 24, 14, 4, 2 and 1 hours prior to detection of remaining plate-bound biotinylated FNfn10-3JCLI4 using streptavidin-horseradish peroxidase and ABTS.\",\n \"Baseline binding activity (100%) was calculated as the amount of biotinylated FNfn10-3JCLI4 which remained bound to the plate following incubation of the well for 24 hours, in the presence of buffer alone (i.e., in the absence of any competitor protein), and the O.D.400 of FNfn10-3JCLI4 bound in the presence of 10 mM EDTA was subtracted as background.\",\n \"All incubations with competitor (background-subtracted) are expressed as a percentage of the O.D.400 of baseline binding (background-subtracted).\",\n \"Error bars reflect the standard deviation between triplicate wells.\",\n \"To examine rate of association of αvβ3 and biotinylated FNfn10-3JCLI4, a 96-well plate coated with 5 μg/ml αvβ3 integrin was blocked with BSA (FIG.\",\n \"5C).\",\n \"0.01 (▴), 0.02 (▪), or 0.05 (♦) μg/ml of biotinylated FNfn10-3JCLI4 was then added to the coated wells, at 240, 30, 10, 3, and 1 minute prior to washing and detection.\",\n \"After detection with streptavidin-horseradish peroxidase/ABTS, the O.D.400 of the plate was recorded.\",\n \"Error bars reflect the standard deviation between triplicate wells.\",\n \"A 96-well plate was coated with αvβ3, αvβ5, αIIbβ3, α1β1, and α5β1 purified integrins, all at 5 μg/ml (FIG.\",\n \"5D).\",\n \"Biotinylated FNfn10-3JCLI4 (dark bars) or FNfn10-WT (light bars) was then added at a concentration of 0.01 μg/ml, and allowed to incubate for 2 hours, followed by washing and detection.\",\n \"Error bars reflect the standard deviation between triplicate wells.\",\n \"In FIGS.\",\n \"5A-D, the experiments shown are all representative of three different experiments that yielded similar results.\",\n \"FIGS.\",\n \"6A-D are graphs illustrating competition ELISAs to determine relative affinities of FNfn10-3JCLI4, echistatin, FNfn10-WT, and GRGDSPK peptide (SEQ ID No: 27).\",\n \"A BSA-blocked 96-well plate which was coated with αvβ3 at 5 μg/ml, after which biotinylated FNfn10-3JCLI4 was then added in the presence or absence of a 0.01, 0.1, 1, 10, or 100-fold molar excess of unlabelled competitor protein, and incubated for two hours.\",\n \"After washing and detection, the O.D.400 of the plate was recorded.\",\n \"Error bars reflect the standard deviation between triplicate wells, and the results shown are representative of three different experiments that yielded similar results.\",\n \"A background subtraction was performed on all data points, by subtracting the measured O.D.400 value that corresponded to binding of biotinylated FNfn10-3JCLI4 in the presence of 10 mM EDTA.\",\n \"All values in the figure are expressed as a percentage of the O.D.400 of biotinylated FNfn10-3JCLI4 binding in the absence of competitor.\",\n \"FIG.\",\n \"6A illustrates the binding competition with unlabelled FNfn10-3JCLI4 (♦) or unlabelled FNfn10-WT (▪).\",\n \"FIG.\",\n \"6B illustrates the binding competition with unlabelled FNfn10-3JCLI4 (▴), echistatin (▪), and GRGDSPK peptide (♦)(SEQ ID No: 27).\",\n \"FIGS.\",\n \"7A-C are graphs illustrating fluorescence activated cell sorting (FACS) analysis of FNfn10-3JCLI4 binding to human cells.\",\n \"Biotinylated FNfn10-3JCLI4 or FNfn10-WT were added to 105 cells at a concentration of 1 μg/ml and incubated for 30 minutes.\",\n \"Bound proteins were then detected with streptavidin-APC, and the mean fluorescence of ten thousand cells per sample was recorded.\",\n \"All experiments were performed three times, with similar results; error bars (FIGS.\",\n \"7B-C) represent the standard deviation of three samples of 105 cells stained separately.\",\n \"FIG.\",\n \"7A is a histogram plot of flow cytometric staining analysis of K562 and K562-αvβ3 cells using FNfn10-3JCLI4.FIG.\",\n \"7B shows mean APC fluorescence for integrin-transfected K562 cells stained with biotinylated FNfn10-3JCLI4 (dark bars) or FNfn10-WT (light bars).\",\n \"To determine the effect of high temperature on FNfn10-3JCLI4 stability, 1 μg/ml 3JCLI4 (♦) or 5 μg/ml LM609 (▪) were incubated for 24 hours in FACS buffer at 4, 20, 37, 50, 65, or 75° C. (FIG.\",\n \"7C).\",\n \"105 K562-αvβ3 cells were pelleted for each sample and re-suspended in the appropriately treated FNfn10-3JCLI4 or LM609, followed by detection as above.\",\n \"FIGS.\",\n \"8A-C illustrate the inhibition of integrin-dependent cell adhesion.\",\n \"Vitronectin (FIGS.\",\n \"8A-B) or fibronectin (FIG.\",\n \"8C), as well as BSA-coated control wells, were incubated with 105 K562 cells, K562-αvβ3 cells, or K562-αIIbβ3 cells for two hours in the presence or absence of the indicated inhibitors.\",\n \"Non-adherent cells were removed by washing, while adherent cells were stained with crystal violet, solubilized, and quantitated by measuring the O.D.570.The background level of crystal violet in control wells was calculated on the basis of measured O.D.570 for BSA-coated wells (no integrin).\",\n \"Error bars represent the standard deviation of triplicate wells, and all experiments were performed three times, with similar results.\",\n \"In FIG.\",\n \"8A, adhesion of binding of K562-αvβ3 cells to vitronectin is shown.\",\n \"Inhibitors added include FNfn10-3JCLI4, FNfn10-WT, and LM609 antibody, at the stated final concentrations.\",\n \"The lane marked “no αvβ3” corresponds to control K562 cells (no αvβ3), while “no VN” denotes cell adherence (K562-αvβ3 cells) measured on control wells (no vitronectin); the lane marked “NT” corresponds to K562-αvβ3 cells which were bound to vitronectin in the absence of any competitor.\",\n \"In FIG.\",\n \"8B, adhesion of K562-αvβ3 cells to vitronectin is shown.\",\n \"Echistatin (▪) and FNfn10-3JCLI4 (♦) were added at final concentrations of 400, 40, and 4 nM, prior to addition of the cells and initiation of the binding assay.\",\n \"Binding is expressed as a percentage of the O.D.570 recorded for K562-αvβ3 adhesion in the absence of competitor, and a background subtraction was performed for all data points, which corresponded to the measured O.D.570 for BSA-coated wells (no integrin).\",\n \"In FIG.\",\n \"8C, adhesion of K562-αIIbβ3 cells to fibronectin is shown.\",\n \"Echistatin (▪) and FNfn10-3JCLI4 (♦) were added at final concentrations of 400, 40, and 4 nM prior to addition of the cells and initiation of the binding assay.\",\n \"Binding is expressed as a percentage of the O.D.570 recorded for K562-αIIbβ3 adhesion in the absence of competitor, and a background subtraction was performed for all data points, which corresponded to the measured O.D.570 for BSA-coated wells (no integrin).\",\n \"DETAILED DESCRIPTION OF THE INVENTION As used herein, “polypeptide monobody” is intended to mean a polypeptide which includes a β-strand domain lacking in disulfide bonds and containing a plurality of β-strands, two or more loop regions each connecting one β-strand to another β-strand, and optionally an N-terminal tail, a C-terminal tail, or both, wherein at least one of the two or more loop regions, the N-terminal tail, or the C-terminal tail is characterized by activity in binding a target protein or molecule.\",\n \"More specifically, such polypeptide monobodies of the present invention can include three or more loop regions or, even more specifically, four or more loop regions.\",\n \"The size of such polypeptide monobodies is preferably less than about 30 kDa, more preferably less than about 20 kDa.\",\n \"The polypeptide monobodies of the present invention are also characterized by specificity for binding to αvβ3 integrin (i.e., enhanced binding specificity for αvβ3 integrin over other integrins, as compared to wild type polypeptide monobodies or wild type fibronectin).\",\n \"To achieve the specificity in their binding to αvβ3 integrin, the amino acid sequence of the polypeptide monobody has been modified relative to the scaffold used for its construction.\",\n \"Scaffolds for formation of a polypeptide monobody should be highly soluble and stable.\",\n \"It is small enough for structural analysis, yet large enough to accommodate multiple binding domains so as to achieve tight binding and/or high specificity for its target.\",\n \"An exemplary scaffold for formation of a polypeptide monobody is the fibronectin type III domain (Fn3).\",\n \"Fibronectin is a large protein which plays essential roles in the formation of extracellular matrix and cell-cell interactions; it consists of many repeats of three types (types I, II, and III) of small domains (Baron et al., 1991).\",\n \"Fn3 itself is the paradigm of a large subfamily (Fn3 family or s-type Ig family) of the immunoglobulin superfamily.\",\n \"The Fn3 family includes cell adhesion molecules, cell surface hormone and cytokine receptors, chaperoning, and carbohydrate-binding domains (for reviews, see Bork & Doolittle, 1992; Jones, 1993; Bork et al., 1994; Campbell & Spitzfaden, 1994; Harpez & Chothia, 1994).\",\n \"Crystallographic studies have revealed that the structure of the DNA binding domains of the transcription factor NF-kB is also closely related to the Fn3 fold (Ghosh et al., 1995; Müller et al., 1995).\",\n \"These proteins are all involved in specific molecular recognition, and in most cases ligand-binding sites are formed by surface loops, suggesting that the Fn3 scaffold is an excellent framework for building specific binding proteins.\",\n \"The 3D structure of Fn3 has been determined by NMR (Main et al., 1992) and by X-ray crystallography (Leahy et al., 1992; Dickinson et al., 1994).\",\n \"The structure is best described as a β-sandwich similar to that of antibody VH domain except that Fn3 has seven β-strands (see FIG.\",\n \"1) instead of nine.\",\n \"There are three loops on each end of Fn3; the positions of the BC, DE, and FG loops approximately correspond to those of CDR 1, 2 and 3 of the VH domain.\",\n \"Fn3 is small (˜94 residues), monomeric, soluble, and stable.\",\n \"It is one of few members of IgSF that do not have disulfide bonds and, therefore, is stable under reducing conditions.\",\n \"Fn3 has been expressed in E. Coli (Aukhil et al., 1993).\",\n \"In addition, 17 Fn3 domains are present just in human fibronectin, providing important information on conserved residues which are often important for the stability and folding (see Main et al., 1992; Dickinson et al., 1994).\",\n \"From sequence analysis, large variations are seen in the BC and FG loops, suggesting that the loops are not crucial to stability.\",\n \"NMR studies have revealed that the FG loop is highly flexible; the flexibility has been implicated for the specific binding of the 10th Fn3 to α5β1 integrin through the Arg-Gly-Asp (RGD) motif In the crystal structure of human growth hormone-receptor complex (de Vos et al., 1992), the second Fn3 domain of the receptor interacts with growth hormone via the FG and BC loops, suggesting it is feasible to build a binding site using the two loops.\",\n \"The tenth type III module of fibronectin has a fold similar to that of immunoglobulin domains, with seven β strands forming two antiparallel β sheets, which pack against each other (FIG.\",\n \"1; Main et al., 1992).\",\n \"The structure of the type H module includes seven β strands, which form a sandwich of two antiparallel sheets, one containing three strands (ABE) and the other four strands (C′CFG) (Williams & Barclay, 1988).\",\n \"The β sheet contains residues Glu-9-Thr-14 (A), Ser-17-Asp-23 (B), and Thr-56-Ser-60 (E).\",\n \"The majority of the conserved residues contribute to the hydrophobic core, with the invariant hydrophobic residues Trp-22 and Try-68 lying toward the N-terminal and C-terminal ends of the core, respectively.\",\n \"The β strands are much less flexible and appear to provide a rigid framework upon which functional, flexible loops can be built.\",\n \"The topology is similar to that of immunoglobulin C domains.\",\n \"Preferred polypeptide monobodies of the present invention are fibronectin type III (Fn3)-derived polypeptide monobodies.\",\n \"Fn3 monobodies include at least two Fn3 β-strand domain sequences with a loop region sequence linked between adjacent β-strand domain sequences and optionally, an N-terminal tail of at least about 2 amino acids, a C-terminal tail of at least about 2 amino acids, or both.\",\n \"The loop region sequence, the N-terminal tail, or the C-terminal tail, or combinations thereof include an amino acid sequence which has binding specificity for αvβ3 integrin.\",\n \"To render a loop region sequence, N-terminal tail, or C-terminal tail capable of binding to αvβ3 integrin, either the loop region sequence, the N-terminal tail, the C-terminal tail, or a combination thereof varies by deletion, insertion, or replacement of at least two amino acids from a corresponding loop region, N-terminal tail, or C-terminal tail in a wild-type or mutant Fn3 scaffold.\",\n \"One preferred wild-type Fn3 scaffold is the tenth Fn3 domain of human fibronectin (FNfn10), which has an amino acid sequence according to SEQ ID No: 2 (FIG.\",\n \"3A).\",\n \"One preferred mutant Fn3 scaffold is the tenth Fn3 domain of human fibronectin which has a modified Asp7, which is replaced by a non-negatively charged amino acid residue (i.e., Asn, Lys, etc.)\",\n \"as shown in FIG.\",\n \"3B (SEQ ID No: 3).\",\n \"As reported in Koide et al.\",\n \"(2001), both of these mutations have the effect of promoting greater stability of the mutant FNfn10 at neutral pH as compared to the wild-type FNfn10.Both the mutant and wild-type FNfn10 are characterized by the same structure, namely seven β-strand domain sequences (designated A through G) and six loop regions (AB loop, BC loop, CD loop, DE loop, EF loop, and FG loop) which connect the seven β-strand domain sequences.\",\n \"In SEQ ID Nos: 2 and 3, the AB loop corresponds to residues 15-16, the BC loop corresponds to residues 22-30, the CD loop corresponds to residues 39-45, the DE loop corresponds to residues 51-55, the EF loop corresponds to residues 60-66, and the FG loop corresponds to residues 76-87.As shown in FIG.\",\n \"1, the BC loop, DE loop, and FG loop are all located at the same end of the polypeptide monobody.\",\n \"According to one particular embodiment, the preferred polypeptide monobodies are mutant or wild-type FNfn10 scaffolds which include a modified FG loop region, which polypeptide monobodies exhibit increased activity in binding to αvβ3 integrin.\",\n \"Such binding is preferably selective of αvβ3 integrin over other integrins.\",\n \"The modified FG loop region preferably includes an amino acid sequence including XRGDWXEX (SEQ ID No: 26), where X is any amino acid residue.\",\n \"The polypeptide monobodies of the present invention can be prepared by recombinant techniques, thereby affording the deletion, insertion, or replacement of at least two amino acids from a corresponding loop region, N-terminal tail, or C-terminal tail in a wild-type or mutant Fn3 scaffold.\",\n \"Deletions can be a deletion of at least two amino acid residues up to substantially all but one amino acid residue appearing in a particular loop region or tail.\",\n \"Insertions can be an insertion of at least two amino acid residues up to about 25 amino acid residues, preferably at least two up to about 15 amino acid residues.\",\n \"Replacements can be replacements of at least two up to substantially all amino acid residues appearing in a particular loop region or tail.\",\n \"The polypeptide monobodies preferably possess an amino acid sequence which is at least 50% homologous to a β-strand domain of the FNfn10.The deletions, insertions, and replacements (relative to wild-type or previously known mutant) on Fn3 scaffolds can be achieved using recombinant techniques beginning with a known nucleotide sequence.\",\n \"A synthetic gene for the tenth Fn3 of human fibronectin (FIG.\",\n \"2) was designed which includes convenient restriction sites for ease of mutagenesis and uses specific codons for high-level protein expression (Gribskov et al., 1984).\",\n \"This gene is substantially identical to the gene disclosed in co-pending U.S. patent application Ser.\",\n \"No.\",\n \"09/096,749 to Koide filed Jun.\",\n \"12, 1998, which is hereby incorporated by reference in its entirety.\",\n \"The gene was assembled as follows: first the gene sequence was divided into five parts with boundaries at designed restriction sites (FIG.\",\n \"2); for each part, a pair of oligonucleotides that code opposite strands and have complementary overlaps of about 15 bases was synthesized; the two oligonucleotides were annealed and single strand regions were filled in using the Klenow fragment of DNA polymerase; the double-stranded oligonucleotide was cloned into the pET3a vector (Novagen) using restriction enzyme sites at the termini of the fragment and its sequence was confirmed by an Applied Biosystems DNA sequencer using the dideoxy termination protocol provided by the manufacturer; and these steps were repeated for each of the five parts to obtain the whole gene.\",\n \"Although this approach takes more time to assemble a gene than the one-step polymerase chain reaction (PCR) method (Sandhu et al., 1992), no mutations occurred in the gene.\",\n \"Mutations would likely have been introduced by the low fidelity replication by Taq polymerase and would have required time-consuming gene-editing.\",\n \"Recombinant DNA manipulations were performed according to Molecular Cloning (Sambrook et al., 1989), unless otherwise stated.\",\n \"To avoid the introduction of mutations during one-step PCR, high fidelity/low error polymerases can be employed as is known in the art.\",\n \"Desired mutations can be introduced to the Fn3 gene using either cassette mutagenesis, oligonucleotide site-directed mutagenesis techniques (Deng & Nickoloff, 1992), or Kunkel mutagenesis (Kunkel et al., 1987).\",\n \"Both cassette mutagenesis and site-directed mutagenesis can be used to prepare specifically desired nucleotide coding sequences.\",\n \"Cassette mutagenesis can be performed using the same protocol for gene construction described above and the double-stranded DNA fragment coding a new sequence can be cloned into a suitable expression vector.\",\n \"Many mutations can be made by combining a newly synthesized strand (coding mutations) and an oligonucleotide used for the gene synthesis.\",\n \"Regardless of the approach utilized to introduce mutations into the monobody nucleotide sequence, sequencing can be performed to confirm that the designed mutations (and no other mutations) were introduced by mutagenesis reactions.\",\n \"In contrast, Kunkel mutagenesis can be utilized to randomly produce a plurality of mutated monobody coding sequences which can be used to prepare a combinatorial library of polypeptide monobodies for screening.\",\n \"Basically, targeted loop regions (or C-terminal or N-terminal tail regions) can be randomized using the NNK codon (N denoting a mixture of A, T, G, C, and K denoting a mixture of G and T) (Kunkel et al., 1987).\",\n \"An exemplary DNA molecule encoding a mutant Fn3 polypeptide monobody is set forth as SEQ ID No: 28 below: atgcaggttt ctgatgttcc gcgtgacctg gaagttgttg ctgcgacccc gactagcctg 60 ctgatcagct gggatgctcc tgcagttacc gtgcgttatt accgtatcac gtacggtgaa 120 accggtggta actccccggt tcaggaattc actgtacctg gttccaagtc tactgctacc 180 atcagcggcc tgaaaccggg tgtcgactat accatcactg tatacgctgt tactnnkcgc 240 ggcgatnnkn nknnknnktc caagccaatc tcgattaact accgtaccgg tggcaggttct 300 ggtggcggt 309 In the above DNA sequence, codons nt 235-237, nt 247-249, nt 250-252, nt 253-255, and nt 256-258 (bold typeface) encode random amino acid residues within the FG loop.\",\n \"According to the preferred embodiments of the present invention, however, codons nt 247-249 and nt 253-255 encode, respectively tryptophan and glutamic acid.\",\n \"The seven codons extending from nt 230-250 encode a seven residue linker which, though not required for formation of a polypeptide monobody of the present invention, is useful in forming fusion proteins as described below.\",\n \"Regardless of the approach used to prepare the nucleic acid molecules encoding the polypeptide monobody, the nucleic acid can be incorporated into host cells using conventional recombinant DNA technology.\",\n \"Generally, this involves inserting the DNA molecule into an expression system to which the DNA molecule is heterologous (i.e., not normally present).\",\n \"The heterologous DNA molecule is inserted into the expression system or vector in sense orientation and correct reading frame.\",\n \"The vector contains the necessary elements (promoters, suppressers, operators, transcription termination sequences, etc.)\",\n \"for the transcription and translation of the inserted protein-coding sequences.\",\n \"Alternatively, a recombinant gene or DNA construct can be prepared prior to its insertion into an expression vector.\",\n \"For example, using conventional recombinant DNA techniques, a promoter-effective DNA molecule can be operably coupled 5′ of a DNA molecule encoding the polypeptide monobody and a transcription termination (i.e., polyadenylation sequence) can be operably coupled 3′ thereof.\",\n \"U.S. Pat.\",\n \"No.\",\n \"4,237,224 to Cohen and Boyer describes the production of expression systems in the form of recombinant plasmids using restriction enzyme cleavage and ligation with DNA ligase.\",\n \"These recombinant plasmids are then introduced by means of transformation and replicated in unicellular cultures including prokaryotic organisms and eukaryotic cells grown in tissue culture.\",\n \"Recombinant molecules can be introduced into cells via transformation, particularly transduction, conjugation, mobilization, or electroporation.\",\n \"The DNA sequences are cloned into the vector using standard cloning procedures in the art, as described by Sambrook et al.\",\n \"(1989).\",\n \"A variety of host-vector systems may be utilized to express the polypeptide monobody or fusion protein which includes a polypeptide monobody.\",\n \"Primarily, the vector system must be compatible with the host cell used.\",\n \"Host-vector systems include but are not limited to the following: bacteria transformed with bacteriophage DNA, plasmid DNA, or cosmid DNA; microorganisms such as yeast containing yeast vectors; and mammalian cell systems infected with virus (e.g., vaccinia virus, adenovirus, etc.).\",\n \"The expression elements of these vectors vary in their strength and specificities.\",\n \"Depending upon the host-vector system utilized, any one of a number of suitable transcription and translation elements can be used.\",\n \"Different genetic signals and processing events control many levels of gene expression (e.g., DNA transcription and messenger RNA (mRNA) translation).\",\n \"Transcription of DNA is dependent upon the presence of a promoter which is a DNA sequence that directs the binding of RNA polymerase and thereby promotes mRNA synthesis.\",\n \"The DNA sequences of eukaryotic promoters differ from those of prokaryotic promoters.\",\n \"Furthermore, eukaryotic promoters and accompanying genetic signals may not be recognized in or may not function in a prokaryotic system and, further, prokaryotic promoters may not be recognized in or may not function in eukaryotic cells.\",\n \"Promoters suitable for use in each of these systems are well known in the art.\",\n \"Similarly, translation of mRNA in prokaryotes depends upon the presence of the proper prokaryotic signals which differ from those of eukaryotes.\",\n \"Efficient translation of mRNA in prokaryotes requires a ribosome binding site called the Shine-Dalgarno (“SD”) sequence on the mRNA.\",\n \"This sequence is a short nucleotide sequence of mRNA that is located before the start codon, usually AUG, which encodes the amino-terminal methionine of the protein.\",\n \"The SD sequences are complementary to the 3′-end of the 16S rRNA (ribosomal RNA) and probably promote binding of mRNA to ribosomes by duplexing with the rRNA to allow correct positioning of the ribosome.\",\n \"For a review on maximizing gene expression, see Roberts & Lauer (1979).\",\n \"In addition, polyadenylation signals suitable for use in a desired host cell can also be employed to effect appropriate translation of the DNA molecule (encoding the polypeptide monobody of the present invention.\",\n \"Polyadenylation signals suitable for use in each of the above-identified systems are well known in the art.\",\n \"Basically, the DNA molecule encoding the polypeptide monobody is ligated to the appropriate promoter and polyadenylation sequences using conventional recombinant techniques, with the promoter being located upstream or 5′ to the DNA molecule and the polyadenylation signal being located downstream or 3′ of the DNA molecule.\",\n \"As a result of ligating the DNA molecule to the promoter and polyadenylation signal sequences, a DNA construct of the present invention is formed.\",\n \"This DNA construct can be cloned directly into a suitable expression vector (e.g., plasmid, viral DNA, etc.)\",\n \"or it can be formed upon cloning the DNA molecule into an empty expression vector which includes an appropriate restriction site intermediate the promoter and polyadenylation signals which are present in the empty vector.\",\n \"Once the DNA molecule encoding the polypeptide monobody has been cloned into an expression system, it is ready to be incorporated into a host cell.\",\n \"Such incorporation can be carried out by the various forms of transformation noted above, depending upon the vector/host cell system.\",\n \"Suitable host cells include, but are not limited to, bacteria, yeast cells, mammalian cells, etc.\",\n \"Upon growing the host cells in a suitable growth medium, the host cells are encouraged to express the polypeptide monobodies of the present invention.\",\n \"In at least several of the various uses of the polypeptide monobodies of the present invention, discussed infra, it is often desirable for the polypeptide monobodies to be produced in substantially purified form, particularly when their administration to a patient is contemplated.\",\n \"Purification can be carried out according to previously reported procedures, which involve metal affinity chromatography for monobodies containing a poly-histidine tag (see Koide et al., 1998).\",\n \"Another aspect of the present invention relates to a polypeptide (i.e., whether in the form of a polypeptide monobody or not) which includes the amino acid sequence RGDWXE (amino acid residues 2-7 of SEQ ID No: 26) and selectively binds αvβ3 integrin over other integrins.\",\n \"Such a polypeptide can be prepared, isolated, and purified according to known procedures.\",\n \"Typically, this involves recombinant expression of the desired polypeptide monobody by a host cell, propagation of the host cells, lysing the host cells, and recovery of supernatant by centrifugation to remove host cell debris.\",\n \"The supernatant can be subjected to sequential ammonium sulfate precipitation.\",\n \"The fraction containing the polypeptide monobodies of the present invention is then subjected to gel filtration in an appropriately sized dextran or polyacrylamide column to separate the polypeptide monobodies.\",\n \"If necessary, the protein fraction may be further purified by HPLC.\",\n \"A further aspect of the present invention relates to fusion proteins which include a polypeptide or polypeptide monobody of the present invention and a second polypeptide linked by peptide bond to the polypeptide or polypeptide monobody.\",\n \"The second polypeptide can be (i) an epitope tag polypeptide, e.g., polyhistidine, (ii) a detectable marker polypeptide, e.g., alkaline phosphatase, (iii) a metal ion-complexing polypeptide, or (iv) a DNA-binding polypeptide, e.g., polylysine.\",\n \"Each of these fusion proteins can be prepared using recombinant techniques described above to form an in-frame gene fusion that can be expressed in appropriate host cells.\",\n \"Thereafter, the fusion protein can isolated and purified using the techniques described above.\",\n \"Once the polypeptides, polypeptide monobodies, or fusion proteins have been obtained in isolated or, more preferably, purified form, they can be used for therapeutic, preventative, or diagnostic purposes as described hereinafter.\",\n \"Because the polypeptides and polypeptide monobodies of the present invention are capable of binding αvβ3 integrin selectively, they can be used for therapeutic, preventative, or diagnostic purposes, or to modify αvβ3 integrin activity.\",\n \"Although references below are made to use of the polypeptide or polypeptide monobody, it should be appreciated that the polypeptide or polypeptide monobody can also be in the form of a fusion protein or conjugate as described elsewhere herein.\",\n \"Thus, one aspect of the present invention relates to a method of treating or preventing an αvβ3 integrin-mediated disease or disorder which includes administering to a patient in need thereof, an effective amount of a polypeptide or polypeptide monobody of the present invention which selectively binds to αvβ3 integrin, thereby treating or preventing the αvβ3 integrin-mediated disease or disorder.\",\n \"Such administration is carried out under conditions which are effective to treat or prevent the αvβ3 integrin-mediated disease or disorder.\",\n \"Typically, this includes administering the polypeptide or polypeptide monobody to the patient in a manner effective to contact one or more cells expressing αvβ3 integrin with a sufficient amount of the polypeptide or polypeptide monobody to inhibit the activity of αvβ3 integrin on such one or more cells.\",\n \"As a result of such inhibition, the αvβ3 integrin-mediated diseases or disorders are treated or prevented.\",\n \"For therapeutic or preventative treatments, the patient can be any mammal which expresses αvβ3 integrin, preferably though not exclusively humans.\",\n \"Integrins are a group of cell surface glycoproteins that mediate cell adhesion and therefore are mediators of cell adhesion interactions that occur in various biological processes.\",\n \"Integrins are heterodimers composed of noncovalently linked α and β polypeptide subunits.\",\n \"Currently at least eleven different α subunits have been identified and at least six different β subunits have been identified.\",\n \"The various α subunits can combine with various β subunits to form distinct integrins.\",\n \"The integrin identified as αvβ3 (also known as the vitronectin receptor) has been identified as an integrin that plays a role in various conditions or disease states including tumor metastasis, solid tumor growth (neoplasia), osteoporosis, Paget's disease, humoral hypercalcemia of malignancy, angiogenesis, including tumor angiogenesis, retinopathy, arthritis, including rheumatoid arthritis, periodontal disease, psoriasis and smooth muscle cell migration (e.g.\",\n \"restenosis).\",\n \"Additionally, it has been found that integrin inhibiting agents would be useful as antivirals, antifungals and antimicrobials.\",\n \"Thus, polypeptides or polypeptide monobodies that selectively bind αvβ3 integrin, thereby inhibiting αvβ3 integrin-mediated conditions, would be beneficial for treating or preventing such conditions.\",\n \"It has been shown that the αvβ3 integrin binds to a number of Arg-Gly-Asp (RGD) containing matrix macromolecules, such as fibrinogen (Bennett et al., 1983), fibronectin (Ginsberg et al., 1983), and von Willebrand factor (Ruggeri et al., 1982).\",\n \"Compounds containing the RGD sequence mimic extracellular matrix ligands so as to bind to cell surface receptors.\",\n \"However, it is also known that RGD peptides in general are non-selective for RGD dependent integrins.\",\n \"For example, most RGD peptides that bind to αvβ3 also bind to αvβ5, αvβ1, and αIIbβIIIa.\",\n \"Antagonism of platelet αIIbβIIIa (also known as the fibrinogen receptor) is known to block platelet aggregation in humans.\",\n \"In order to avoid bleeding side-effects when treating the conditions of disease states associated with the integrin αvβ3, it would be beneficial to develop agents which selectively bind to αvβ3 as opposed to αIIbβIIIa as well as other integrins.\",\n \"Polypeptides or polypeptide monobodies selective for αvβ3 offer such an advantage.\",\n \"As noted above, integrins are believed to be involved in, among other biological processes, tumor metastasis and neoplasia.\",\n \"Tumor cell invasion occurs by a three step process: 1) tumor cell attachment to extracellular matrix; 2) proteolytic dissolution of the matrix; and 3) movement of the cells through the dissolved barrier.\",\n \"This process can occur repeatedly and can result in metastases at sites distant from the original tumor.\",\n \"Studies have shown (Humphries et al., 1988), that RGD-like compounds can interfere with experimental metastasis wherein tumor cells are injected directly into the blood.\",\n \"In addition, Seftor et al.\",\n \"(1992) have shown that the αvβ3 integrin has a biological function in melanoma cell invasion.\",\n \"Montgomery et al., (1994) demonstrated that the integrin αvβ3 expressed on human melanoma cells promotes a survival signal, protecting the cells from apoptosis.\",\n \"Accordingly, mediation of the tumor cell metastatic pathway by interference with the αvβ3 integrin cell adhesion receptor to impede tumor metastasis would be beneficial.\",\n \"Brooks et al.\",\n \"(1994a) have demonstrated that antagonists of αvβ3 provide a therapeutic approach for the treatment of neoplasia (inhibition of solid tumor growth) since systemic administration of αvβ3 antagonists causes dramatic regression of various histologically distinct human tumors.\",\n \"The adhesion receptor integrin αvβ3 was also identified as a marker of angiogenic blood vessels in chick and man and plays a critical role in angiogenesis or neovascularization (Brooks et al., 1995).\",\n \"Angiogenesis is characterized by the invasion, migration and proliferation of smooth muscle and endothelial cells.\",\n \"Antagonists of αvβ3 inhibit this process by selectively promoting apoptosis of cells in neovasculature.\",\n \"The growth of new blood vessels, or angiogenesis, also contributes to pathological conditions such as diabetic retinopathy (Adonis et al., 1994), rheumatoid arthritis (Peacock et al., 1992) and osteoarthritis (Ondrick et al., 1992).\",\n \"Therefore, αvβ3 antagonists are useful targets for treating or preventing such conditions associated with neovascularization (Brooks et al., 1994b).\",\n \"Because angiogenesis occurs normally in the female reproductive organs, antagonists of αvβ3 would be useful in controlling fertility.\",\n \"It has also been reported that the cell surface receptor αvβ3 is the major integrin on osteoclasts responsible for attachment to bone.\",\n \"Osteoclasts cause bone resorption and when such bone resorbing activity exceeds bone forming activity it results in osteoporosis (a loss of bone), which leads to an increased number of bone fractures, incapacitation and increased mortality.\",\n \"Antagonists of αvβ3 have been shown to be potent inhibitors of osteoclastic activity both in vitro (Sato et al., 1990) and in vivo (Fisher et al., 1993).\",\n \"Antagonism of αvβ3 leads to decreased bone resorption and therefore restores a normal balance of bone forming and resorbing activity.\",\n \"Thus, it would be beneficial to provide antagonists of osteoclast αvβ3 which are effective inhibitors of bone resorption and, therefore, are useful in the treatment or prevention of osteoporosis.\",\n \"The role of the αvβ3 integrin in smooth muscle cell migration also makes it a therapeutic target for prevention or inhibition of neointimal hyperplasia which is a leading cause of restenosis after vascular procedures (Choi et al., 1994).\",\n \"Prevention or inhibition of neointimal hyperplasia (restenosis) by the polypeptides or polypeptide monobodies of the present invention will be beneficial.\",\n \"White (1993) has reported that adenovirus uses αvβ3 for entering host cells.\",\n \"The integrin appears to be required for endocytosis of the virus particle and may be required for penetration of the viral genome into the host cell.\",\n \"Thus, polypeptides or polypeptide monobodies of the present invention which inhibit αvβ3 will be useful as antiviral agents.\",\n \"As a result of the present invention, polypeptides or polypeptide monobodies of the present invention (with αvβ3 selective binding activity) can be used for blocking αvβ3-mediated events such as cell adhesion, osteoclast-mediated bone resorption, restenosis, ocular neovascularization and growth of hemangiomas, as well as neoplastic cell or tumor growth and dissemination.\",\n \"The inventive polypeptide or polypeptide monobodies can also be used for mediated targeting and delivery of therapeutics for disrupting or killing αvβ3 bearing neoplasms and tumor-related vascular beds.\",\n \"In addition, the inventive polypeptide or polypeptide monobodies can be used for visualization or imaging of αvβ3 bearing neoplasms or tumor related vascular beds, for example, by MRI or immunoscintigraphy.\",\n \"In addition, these polypeptide or polypeptide monobodies can be used to detect αvβ3 in solution, in frozen tissue sections, and on the surface of cells; therefore, these polypeptides or polypeptide monobodies may be used for the detection and characterization of αvβ3-bearing tumor and endothelial cells in human malignancies.\",\n \"Accordingly, the polypeptides or polypeptide monobodies of the present invention may be used for identification of tumor vasculature.\",\n \"Since the integrin αvβ3 is minimally expressed on resting or normal blood vessels, but is significantly up-regulated on vascular cells within human tumors (Brooks et al., 1994a; Brooks et al., 1994b; Brooks et al., 1995), αvβ3 may be considered a marker of human tumor-associated blood vessels and tumor growth (Brooks et al., 1995).\",\n \"Consequently, polypeptides or polypeptide monobodies that recognize αvβ3 may be used as a diagnostic agent to identify tumor-related blood vessels or vascular beds using conventional immunohistochemical techniques.\",\n \"Further, since the integrin αvβ3 is a marker of tumor-associated blood vessels and recent findings indicate that blood vessel density is a prognostic indicator of cancer and disease status (Weidner, 1993), polypeptides or polypeptide monobodies that recognize αvβ3 may be used as a component of a diagnostic device or technique to measure tumor-related blood vessel abundance or density and to define disease prognosis.\",\n \"Specifically, histologic sections from fresh-frozen or formalin-fixed paraffin embedded tumor tissue are immunostained with αvβ3-specific polypeptides or polypeptide monobodies, i.e., at concentration of 0.1-50 μg/ml, using techniques and procedures known to those skilled in the art.\",\n \"αvβ3-specific polypeptides or polypeptide monobodies bound to vascular endothelial cells in the tissue can be visualized using a polypeptide or polypeptide monobody fusion or a secondary agent that permits localization of the αvβ3-specific polypeptides or polypeptide monobodies bound within the tissue section.\",\n \"For fusions, the αvβ3-specific polypeptides or polypeptide monobodies can be fused with alkaline phosphatase, both of which retain their functionality.\",\n \"The alkaline phosphatase, which is integral with the functional polypeptide or polypeptide monobody portion, can be identified in accordance with known procedures.\",\n \"The secondary agent is typically an antibody that reacts or binds with the αvβ3-specific polypeptides or polypeptide monobodies and has an attached reporter molecule, e.g., an anti-polypeptide or polypeptide monobody antibody with an attached enzyme or fluorescent marker.\",\n \"The polypeptide or polypeptide monobody, when used with secondary agents, is typically provided with an epitope tag fusion, the epitope portion being recognized by the anti-polypeptide or polypeptide monobody antibody.\",\n \"The number and area of immunostained blood vessels in the tissue can then be counted by microscopic techniques, familiar to those skilled in the art, to assess tumor blood vessel density or abundance.\",\n \"The fusion proteins of the present invention which include a DNA binding polypeptide (e.g., polylysine amino acid sequence) can also be used to deliver a therapeutic DNA molecule into a cell which expresses an integrin targeted by the polypeptide or polypeptide monobody of the present invention, such as the αvβ3 integrin.\",\n \"Polylysine has long been known as a DNA-binding motif because of its role in histone-mediated DNA condensation.\",\n \"This property of polylysine to bind DNA was utilized to deliver the DNA to mammalian cells (Wu & Wu, 1987; Wu & Wu, 1988).\",\n \"The technology was utilized by Zhou et al to develop lipophilic polylysines which were in part the basis of present lipofection delivery of DNA (Zhou et al., 1991).\",\n \"Polylysines conjugated by peptide bond to integrin-binding peptides have been described which effectively deliver DNA to cells (Harbottle et al., 1998).\",\n \"Polylysine DNA has also been used to deliver DNA to endothielial cells, a potential target for αvβ3-selective gene delivery (Trubetskoy et al., 1992).\",\n \"Thus, a fusion protein, as described above, is first combined with the DNA to be delivered under conditions which are effective to allow the DNA binding peptide, such as polylysine, to bind the DNA.\",\n \"Thereafter, the complex formed upon binding of the fusion protein to the DNA, is administered to a patient under conditions effective to deliver the DNA into the cells which express the integrin, e.g., αvβ3, targeted by the fusion protein.\",\n \"In addition to its administration alone, a number of known delivery techniques can be utilized for the delivery, into cells, of either the polypeptide monobodies themselves or nucleic acid molecules which encode them.\",\n \"Regardless of the particular method of the present invention which is practiced, when it is desirable to contact a cell (i.e., to be treated) with a polypeptide or polypeptide monobody of the present invention, or its encoding nucleic acid, it is preferred that the contacting be carried out by delivery of the polypeptide monobody or its encoding nucleic acid to the cell.\",\n \"One approach for delivering the polypeptide or polypeptide monobody to the cell or its encoding nucleic acid into cells involves the use of liposomes.\",\n \"Basically, this involves providing the polypeptide or polypeptide monobody or its encoding nucleic acid to be delivered, and then contacting the target cell with the liposome under conditions effective for delivery of the polypeptide or polypeptide monobody or nucleic acid into the cell.\",\n \"Liposomes are vesicles comprised of one or more concentrically ordered lipid bilayers which encapsulate an aqueous phase.\",\n \"They are normally not leaky, but can become leaky if a hole or pore occurs in the membrane, if the membrane is dissolved or degrades, or if the membrane temperature is increased to the phase transition temperature.\",\n \"Current methods of drug delivery via liposomes require that the liposome carrier ultimately become permeable and release the encapsulated drug at the target site.\",\n \"This can be accomplished, for example, in a passive manner wherein the liposome bilayer degrades over time through the action of various agents in the body.\",\n \"Every liposome composition will have a characteristic half-life in the circulation or at other sites in the body and, thus, by controlling the half-life of the liposome composition, the rate at which the bilayer degrades can be somewhat regulated.\",\n \"In contrast to passive drug release, active drug release involves using an agent to induce a permeability change in the liposome vesicle.\",\n \"Liposome membranes can be constructed so that they become destabilized when the environment becomes acidic near the liposome membrane (Wang & Huang, 1987).\",\n \"When liposomes are endocytosed by a target cell, for example, they can be routed to acidic endosomes which will destabilize the liposome and result in drug release.\",\n \"Alternatively, the liposome membrane can be chemically modified such that an enzyme is placed as a coating on the membrane which slowly destabilizes the liposome.\",\n \"Since control of drug release depends on the concentration of enzyme initially placed in the membrane, there is no real effective way to modulate or alter drug release to achieve “on demand” drug delivery.\",\n \"The same problem exists for pH-sensitive liposomes in that as soon as the liposome vesicle comes into contact with a target cell, it will be engulfed and a drop in pH will lead to drug release.\",\n \"This liposome delivery system can also be made to accumulate at a target organ, tissue, or cell via active targeting (e.g., by incorporating an antibody or hormone on the surface of the liposomal vehicle).\",\n \"This can be achieved according to known methods.\",\n \"Different types of liposomes can be prepared according to Bangham et al.\",\n \"(1965); U.S. Pat.\",\n \"No.\",\n \"5,653,996 to Hsu et al.\",\n \"; U.S. Pat.\",\n \"No.\",\n \"5,643,599 to Lee et al.\",\n \"; U.S. Pat.\",\n \"No.\",\n \"5,885,613 to Holland et al.\",\n \"; U.S. Pat.\",\n \"No.\",\n \"5,631,237 to Dzau et al.\",\n \"; and U.S. Pat.\",\n \"No.\",\n \"5,059,421 to Loughrey et al., as well as any other approach demonstrated in the art.\",\n \"From the foregoing, it should be apparent that the polypeptide or polypeptide monobody can either be delivered in a liposome or, when incorporated into the lipid bilayer(s), used as a means for targeting a liposome to αvβ3 integrin-expressing cells.\",\n \"An alternative approach for delivery of polypeptide monobodies involves the conjugation of the desired polypeptide or polypeptide monobody to a polymer that is stabilized to avoid enzymatic degradation of the conjugated polypeptide or polypeptide monobody.\",\n \"Conjugated proteins or polypeptides of this type are described in U.S. Pat.\",\n \"No.\",\n \"5,681,811 to Ekwuribe.\",\n \"When it is desirable to achieve heterologous expression of a desirable polypeptide or polypeptide monobody of the present invention in a target cell, DNA molecules encoding the polypeptide or polypeptide monobody can be delivered into the cell.\",\n \"Basically, this includes providing a nucleic acid molecule encoding the polypeptide or polypeptide monobody and then introducing the nucleic acid molecule into the cell under conditions effective to express the polypeptide monobody in the cell.\",\n \"Preferably, this is achieved by inserting the nucleic acid molecule into an expression vector before it is introduced into the cell.\",\n \"For intracellular expression in this manner, use of polypeptide monobodies is preferred because such monobodies are enzymatically stable within cells.\",\n \"When transforming mammalian cells for heterologous expression of a polypeptide or polypeptide monobody of the present invention, an adenovirus vector 5 can be employed.\",\n \"Adenovirus gene delivery vehicles can be readily prepared and utilized given the disclosure provided in Berkner (1988) and Rosenfeld et al.\",\n \"(1991).\",\n \"Adeno-associated viral gene delivery vehicles can be constructed and used to deliver a gene to cells.\",\n \"The use of adeno-associated viral gene delivery vehicles in vivo is described in Flotte et al.\",\n \"(1993) and Kaplitt et al.\",\n \"(1994).\",\n \"Additional types of adenovirus vectors are described in U.S. Pat.\",\n \"No.\",\n \"6,057,155 to Wickham et al.\",\n \"; U.S. Pat.\",\n \"No.\",\n \"6,033,908 to Bout et al.\",\n \"; U.S. Pat.\",\n \"No.\",\n \"6,001,557 to Wilson et al.\",\n \"; U.S. Pat.\",\n \"No.\",\n \"5,994,132 to Chamberlain et al.\",\n \"; U.S. Pat.\",\n \"No.\",\n \"5,981,225 to Kochanek et al.\",\n \"; U.S. Pat.\",\n \"No.\",\n \"5,885,808 to Spooner et al.\",\n \"; and U.S. Pat.\",\n \"No.\",\n \"5,871,727 to Curiel.\",\n \"Retroviral vectors which have been modified to form infective transformation systems can also be used to deliver nucleic acid encoding a desired polypeptide monobody into a target cell.\",\n \"One such type of retroviral vector is disclosed in U.S. Pat.\",\n \"No.\",\n \"5,849,586 to Kriegler et al.\",\n \"Regardless of the type of infective transformation system employed, it should be targeted for delivery of the nucleic acid to a specific cell type.\",\n \"For example, for delivery of the nucleic acid into tumor cells, a high titer of the infective transformation system can be injected directly within the tumor site so as to enhance the likelihood of tumor cell infection.\",\n \"The infected cells will then express the desired polypeptide or polypeptide monobody, allowing the polypeptide or polypeptide monobody to bind to the αvβ3 integrin.\",\n \"Whether the polypeptides or polypeptide monobodies of the present invention (or encoding nucleic acids) are administered alone or in combination with pharmaceutically or physiologically acceptable carriers, excipients, or stabilizers, or in solid or liquid form such as, tablets, capsules, powders, solutions, suspensions, or emulsions, they can be administered orally, parenterally, subcutaneously, intravenously, intramuscularly, intraperitoneally, by intranasal instillation, by intracavitary or intravesical instillation, intraocularly, intraarterially, intralesionally, or by application to mucous membranes, such as, that of the nose, throat, and bronchial tubes.\",\n \"For most therapeutic purposes, the polypeptides or polypeptide monobodies (or encoding nucleic acids) can be administered intravenously.\",\n \"For injectable dosages, solutions or suspensions of these materials can be prepared in a physiologically acceptable diluent with a pharmaceutical carrier.\",\n \"Such carriers include sterile liquids, such as water and oils, with or without the addition of a surfactant and other pharmaceutically and physiologically acceptable carrier, including adjuvants, excipients or stabilizers.\",\n \"Illustrative oils are those of petroleum, animal, vegetable, or synthetic origin, for example, peanut oil, soybean oil, or mineral oil.\",\n \"In general, water, saline, aqueous dextrose and related sugar solution, and glycols, such as propylene glycol or polyethylene glycol, are preferred liquid carriers, particularly for injectable solutions.\",\n \"For use as aerosols, the polypeptide monobodies or nucleic acids in solution or suspension may be packaged in a pressurized aerosol container together with suitable propellants, for example, hydrocarbon propellants like propane, butane, or isobutane with conventional adjuvants.\",\n \"The materials of the present invention also may be administered in a non-pressurized form such as in a nebulizer or atomizer.\",\n \"Dosages to be administered can be determined according to known procedures, including those which balance both drug efficacy and degree of side effects.\",\n \"From the foregoing, it should be appreciated that therapeutic methods and compositions are contemplated for inhibiting integrins, particularly αvβ3 integrin.\",\n \"These methods are useful to inhibit cell attachment and migration mediated by the integrins and find application in a wide variety of cell types, tissues and systems where attachment of cells is not desired.\",\n \"Thus, in general, the invention contemplates a method for inhibiting an integrin that binds a specific ligand by contacting the integrin with an inhibiting amount of an integrin-inhibiting polypeptide or polypeptide monobody (or fusion protein/conjugate containing them) of the present invention.\",\n \"Typically, the method is practiced on cells expressing the integrin on the surface of the cell, so the contacting occurs by exposing the cells to the integrin-inhibiting polypeptides or polypeptide monobodies.\",\n \"An inhibiting amount of integrin-inhibiting polypeptide or polypeptide monobody is an amount sufficient to produce the desired result, namely to inhibit the integrin to a degree sufficient to reduce the adhesion of the cell expressing the integrin, and typically depends on the amount of integrin to be contacted.\",\n \"In preferred embodiments, whether the method is practiced in vitro or in vivo, an inhibiting amount is an amount sufficient to provide at least about one molar equivalent of integrin-inhibiting polypeptide or polypeptide monobody per molar equivalent of integrin to be inhibited.\",\n \"This amount is referred to as a stoichiometric amount of integrin-inhibiting polypeptide or polypeptide monobody.\",\n \"Although polypeptide or polypeptide monobody affinity is typically sufficient for an integrin-inhibiting polypeptide or polypeptide monobody to react stoichiometrically in dilute solutions, it is preferred that an inhibiting amount is in the range of about 100 nanomolar (nM) to 1 millimolar (mM), preferably in the range of 1 to 100 micromolar (μM), more preferably about 1 to 10 μM.\",\n \"When an integrin-inhibiting method is practiced in vitro, a liquid sample containing integrin, and preferably a physiological fluid containing cells that express cell surface integrin, are admixed with an inhibiting amount of an integrin-inhibiting polypeptide or polypeptide monobody of the present invention to form a complex.\",\n \"The complex is maintained under biological conditions compatible with the formation of an reaction product and also compatible, if required, with cell viability for a time period sufficient for the integrin-inhibiting polypeptide or polypeptide monobody to react with the integrin and, when reacted, inhibit the integrin.\",\n \"When the integrin-inhibiting method is practiced in vivo, an inhibiting amount of a polypeptide or polypeptide monobody composition containing a physiologically tolerable diluent and integrin-inhibiting polypeptide or polypeptide monobody molecules as described herein is administered to a mammal, e.g.\",\n \"a human, and the mammal is maintained for a sufficient period to allow the monobody molecules to react with any integrin present and form an inhibiting reaction product.\",\n \"Suitable routes of administration are envisioned as described above, preferably intravenous, intraperitoneal, intra muscular, intrathecal or subcutaneous.\",\n \"The integrin-inhibiting polypeptide or polypeptide monobody may be combined with other pharmaceutical compositions and/or excipients.\",\n \"For example, αvβ3 integrin-inhibiting polypeptide or polypeptide monobody may be co-administered or added to established anti-cancer chemotherapeutic or biotherapeutic regimens.\",\n \"Normal physiologic saline is a preferred excipient or vehicle for administration of the polypeptide or polypeptide monobody.\",\n \"This may include, but is not limited to, combining or co-administering αvβ3 integrin-inhibiting polypeptide or polypeptide monobody with cytotoxic drugs, combinations of cytotoxic drugs or with immune stimulating drugs, such as interleukins or their derivatives or with hematopoietic factors and their derivatives.\",\n \"For example, αvβ3 monobody may be co-administered or added to therapeutic regimens for the use IL-1, IL-2, IL-4, IL-6, IL-12, IL-15, TNF, α, β, γ, interferons and M-CSF, or combinations of these agents or their derivatives, in biologic therapy of cancer.\",\n \"Further, αvβ3 integrin-inhibiting polypeptide or polypeptide monobody may be co-administered or added to therapeutic regimens for the use of G-CSF, M-CSF, IL-3 and erythropoietin, or combinations of these agents or their derivatives, in biologic therapy of cancer.\",\n \"In addition to its use alone, the integrin-inhibiting polypeptide or polypeptide monobody may also be conjugated to a chemotherapeutic agent to afford a conjugate that can be administered alone or in combination with other anti-cancer drugs or immune-enhancing drugs of the type described above.\",\n \"Conjugates can be formed using known techniques, although typically the conjugate can be formed by linking the chemotherapeutic agent directly or indirectly, via covalent bond, to the C-terminal or N-terminal regions of the monobody.\",\n \"For purposes of imaging cancerous or pre-cancerous lesions to assess the degree of angiogenesis and, therefore, the likelihood of metastatic activity, the polypeptide or polypeptide monobody of the present invention can be formed as a fusion protein including a metal ion-complexing polypeptide or formed as a conjugate including a contrasting agent or chelating agent.\",\n \"Suitable metal ion-complexing polypeptides include, without limitation, polyhistidine.\",\n \"The fusion protein-metal ion complex can be used to identify integrin containing tissues, particularly αvβ3 integrin containing tissues, i.e., where angiogenesis or neoplasia may be occurring.\",\n \"The complexation of the metal ions to the fusion protein, and specifically to the metal ion-complexing polypeptide or chelating agent, is achieved by mixing appropriate amounts of the fusion protein with the metal ion.\",\n \"This is preferably done in solution, with the solution including an appropriate buffer.\",\n \"In one approach, the metal ion is, when mixed with the fusion protein, already in the oxidation state required for complexing to the metal ion-complexing polypeptide.\",\n \"Some metal ions are complexed in their most stable oxidation state, such as ionic forms of calcium, potassium, indium, manganese, copper, zinc, cobalt and other metals.\",\n \"In another approach, the metal ions must be reduced to a lower oxidation state in order to be complexed to the metal ion-complexing polypeptide.\",\n \"This is true of ferrous, ferric, stannous, stannic, technetiumoxo[V], pertechnetate, rheniumoxo[V], perrhenate and other similar metal ions.\",\n \"Thus, for example, both perrhenate and pertechnetate must be reduced to a lower oxidation state prior to complexing.\",\n \"Reduction may be performed prior to mixing with the polypeptide, simultaneously with mixing with the fusion protein, or subsequent to mixing with the fusion protein.\",\n \"Any means of reduction of metal ions to the desired oxidation state known to the art may be employed.\",\n \"For example, perrhenate or pertechnetate may be reduced by use of stannous ions, dithionite or other means.\",\n \"The stannous or dithionite metal ion reducing agent may be mixed with the metal ion to be reduced either prior or subsequent to addition of the metal ion to the fusion protein, or the reducing agent may be in solution with the fusion protein when the metal ion to be reduced, and subsequently complexed to the fusion protein, is added to the solution.\",\n \"The stoichiometric ratios between the fusion protein and the metal ion in the labeling or complexing step can be varied depending on the application.\",\n \"For example, in the case of radiometal complexation, the ratio of radiometal ions to fusion protein molecules can be varied from less than 1:2 to 1:1000 or higher, without generating substantial radiochemical impurities.\",\n \"In other applications, the ratio of metal ions to fusion protein molecules can range from at least 1000:1 to 1:1000 or higher.\",\n \"When the concentration of metal ions is higher than the concentration of fusion protein molecules, all or virtually all of the fusion protein molecules will be complexed to metal ions.\",\n \"Either radioactive or non-radioactive metal ions can be employed.\",\n \"Where a specific diagnostic or therapeutic advantage can be obtained from use of a radioisotope, then a radioactive metal ion is employed.\",\n \"Where the diagnostic or therapeutic utility is obtained from the conformationally constrained biological-function domain, a non-radioactive metal ion is employed.\",\n \"It is possible and contemplated, and may for certain applications be advantageous, to employ different metal ions with the same conjugate product of this invention.\",\n \"Suitable contrasting agents include, without limitation, a dextran-coated superparamagnetic iron oxide (SPIO) particle (Reimer et al., 1991; Schaffer et al., 1993; Reimer et al., 1994; Small et al., 1994; Kresse et al., 1998; Suwa et al., 1998), a lipid vesicle or other similar particle with Gd3+ attached to the surface (Kabalka et al., 1991; Tilcock et al., 1992; Sipkins et al., 1998), and poly-lysine with Gd3+ attached to the amino groups (Kayyem et al., 1995; Vera et al., 1995).\",\n \"Like the fusion proteins containing a metal ion complexing agent, the conjugates containing a contrasting agent can also be used to identify integrin containing tissues, particularly αvβ3 integrin containing tissues, i.e., where angiogenesis or neoplasia may be occurring.\",\n \"Another type of conjugate includes an organic chelating agent (e.g., diethylenetriamine pentaacetic acid) conjugated to a polypeptide monobody of the present invention which has been modified at, e.g., its C-terminus to include a cysteine residue which will form a stable covalent bond with the chelating agent.\",\n \"Thereafter, the conjugate can be exposed to a suitable metal ion of the type described above in a manner which is effective to allow the conjugated diethylenetriamine pentaacetic acid moiety to complex with the metal ions.\",\n \"The fusion proteins or conjugates, and the methods of this invention can also be applied to diagnostic agents for use in positron emission tomography (PET) and magnetic resonance imaging (MRI).\",\n \"For use as a PET agent, a fusion protein or conjugate is complexed with one of the various positron emitting metal ions, such as 51Mn, 52Fe, 60Cu, 68Ga, 72As, 94mTc, 110In, and isotopes of At.\",\n \"For MRI applications, the complexing metal ion is paramagnetic, such as Mn, Gd, Fe, or Dy.\",\n \"EXAMPLES The following Examples are intended to be illustrative and in no way are intended to limit the scope of the present invention.\",\n \"Materials & Methods The following materials and methods were used in the experiments set forth in Examples 1-5.General reagents: All chemicals, unless otherwise specified, were obtained from Sigma (St. Louis, Mo.).\",\n \"Purified integrins, cell adhesion strips, and LM609 were purchased from Chemicon (Temecula, Calif.).\",\n \"Echistatin and GRGDSPK peptide (SEQ ID No: 26) were obtained from Bachem (Torrence, Calif.).\",\n \"Cell Culture: K562, K562-αvβ3, K562-αIIbβ3, K562-αvβ5, K562-α4β1, and K562-α4β7 cells were a gift of Dr. S. Blystone (Upstate Medical University, Syracuse, N.Y.) (Blystone et al., 1994).\",\n \"K562 cells were maintained in Iscove's Modified Dulbecco's Medium (IMDM) (Gibco, Invitrogen, Grand Island, N.Y.) supplemented with 10% fetal bovine serum (Sigma), 0.5 U/L penicillin-streptomycin, and 2 mM L-Glutamine (Gibco).\",\n \"K562-αvβ3, K562-αIIbβ3, K562-αvβ5, K562-α4β1, and K562-α4β7 cells were maintained in the same media containing 500 μg/ml G418 (GENETICIN) (Gibco).\",\n \"Construction of phage vectors: A phage-display vector for FNfn10, designated JCFN, was constructed by cloning the FNfn10 gene (Koide et al., 1998) in the modified M13 vector JC-M13-88 (Chappel et al., 1998).\",\n \"The FNfn10 gene was amplified from pAS38 (Koide et al., 1998) using the following oligonucleotides: FN3JCM8Nhe (SEQ ID No:29) cctagctagc gtagctcagg ccatgcaggt ttctgatgtt c 41 and FN3JCM8Hin (SEQ ID No:30) ggccaagctt gaccgccacc agaaccgcca ccggtacggt ag 42 digested with the restriction enzymes NheI and HindIII, respectively, and subcloned in JC-M13-88 using the same restriction enzyme sites so that the FNfn10 gene is placed in-frame between the OmpA signal sequence and M13 geneVIII.\",\n \"It was confirmed by ELISA that the JCFN phages prepared using E. coli XL-1blue displayed FNfn10 on the phage surface.\",\n \"Purified FNfn10 clones were expressed with an N-terminal histag and a C-terminal GKKGK (SEQ ID No: 31) tag.\",\n \"The GKKGK (SEQ ID No: 31) tag was incorporated to increase the solubility (Koide et al., 1998) and to serve as a preferred biotinylation site.\",\n \"FNfn10 genes were amplified using the following oligonucleotides: FN1F2 (SEQ ID No:32) cgggatccca tatgcaggtt tctgatgttc cgcgtgacct ggaagttgtt gctgcgacc 59 and FN3GKKGK (SEQ ID No:33) ccgactcgag ttactattta ccttttttac cggtacggta gttaatcgag 50 which were digested with the restriction enzymes NdeI and XhoI, and ligated with pAS45 (Koide et al., 1998) digested with the same restriction enzymes.\",\n \"The expression vector for the wild type protein, pAS54, contained an Arg6 to Thr mutation that had been introduced to remove a secondary Thrombin site (Koide et al., 1998).\",\n \"Construction of the FNfn10 library: A FNfn10 library, JCFN-RGD, was constructed in such a way that the FG loop has an XRGDXXXX sequence where X stands for any amino acids (residue #77-84; residue numbering is according to FIG.\",\n \"2(a) of Koide et al.\",\n \"(1998)).\",\n \"Mutagenesis on the JCFN template was performed according to Kunkel's method (Kunkel, 1985) using an oligonucleotide, designated JCFNFGRGD, as follows: JCFNFGRGD (SEQ ID No: 34) gttaatcgag attggcttgg arnnnmnnmnn mnnatcgccg cgmnnagtaa cagcgtatac 60 where N is a mixture of A, G, C and T, and M is a mixture of A and C).\",\n \"E. coli SS320 was electro-transformed with the mutagenesis mixture, resulting in a phage library containing 1.5×109 independent clones.\",\n \"The phages were re-amplified in E. coli XL1-blue with various concentrations of isopropyl β-D-thiogalactopyranoside (IPTG).\",\n \"αvβ3 biopanning using the JCFN-RGD library: Integrin biopanning was performed similarly to the method previously described (Koivunen et al., 1994).\",\n \"Purified αvβ3 (5 μg/ml in TBS buffer containing 2 mM CaCl2) was bound to a single microtiter well of a 96-well plate.\",\n \"The well was rinsed 3 times with sterile water containing 2 mM CaCl2 and blocked with 5% phage blocking reagent (Novagen, Darmstadt, Germany) for one hour at room temperature.\",\n \"The plate was rinsed 5 times with TBS buffer containing 0.1% Tween-20 and 2 mM CaCl2, then incubated with JCFN-RGD library diluted in 5% blocking reagent for one hour.\",\n \"The plate was washed ten times with TBS buffer containing 0.1% Tween 20 and 2 mM CaCl2 and bound phage were eluted with 0.2 M glycine-HCl (pH 2.2) containing 1 mg/ml BSA for 10 minutes at room temperature with gentle agitation and neutralized with 15% (v/v) of 1 M Tris-HCl (pH 9.1).\",\n \"The phage were amplified using the E. coli ER2738 host strain (New England Biolabs, Beverly, Mass.)\",\n \"in the presence of 1 mM, 0.1 mM or with no IPTG and purified by precipitating with 20% (w/v) polyethylene glycol-8000, 2.5 M NaCl and resuspending in PBS containing 20% (v/v) glycerol.\",\n \"The biopanning and amplification of eluted phage were repeated for two more rounds.\",\n \"Upon completion of three rounds of biopanning, individual phage clones were amplified and the genomes were purified by phenol extraction.\",\n \"Polymerase chain reaction was performed to amplify the FNfn10 domain using the following primers: SFN3F (SEQ ID No:35) agctcaattg gtccggtgga ggttctgatg ttccgcgtga cctg 44 and SFN3R (SEQ ID No:36) agctaagctt ttaggtacgg tagttaatcg agat 34 The DNA was also sequenced using the SFN3F primer.\",\n \"Protein preparation: The expression of FNfn10 proteins was performed as previously described (Koide et al., 1998) except that the proteins were further biotinylated.\",\n \"After biotinylation, the protein-containing solution was applied to a nickel affinity column and unbound materials were washed off; the column was then equilibrated with 50 mM sodium phosphate buffer (pH 8.0) containing 500 mM sodium chloride.\",\n \"D-biotinoyl-ε-aminocaptoic acid N-hydroxysuccinimide ester (726 μM) (Boehringer Mannheim, Mannheim, Germany) in the same buffer was applied to the column, and the column was incubated for an hour at room temperature to perform biotinylation.\",\n \"This biotinylation process was performed a total of three times.\",\n \"Then the column was washed with 20 mM Tris-HCl buffer (pH 8.0) containing 500 mM sodium chloride, and the biotinylated FNfn10 was eluted with the buffer containing 500 mM imidazole.\",\n \"The proteins purified in this manner were >90% pure as judged by SDS-PAGE and reverse phase chromatography.\",\n \"Protein concentrations for subsequent analyses were determined by Bradford assay and confirmed by SDS-PAGE analysis and Coomassie Brilliant Blue staining.\",\n \"Biotinylation was confirmed by Western analysis with streptavidin-horseradish peroxidase (Oncogene, Cambridge, Mass.).\",\n \"ELISA detection of biotinylated FNfn10 clones: Purified integrin was bound to a flat-bottom, high-binding EIA/RIA plate (Corning Costar, Corning, N.Y.) at a concentration of 5 μg/ml in TBS for 12-16 hours at 4° C. The plate was washed once with TBS and blocked for 2 hours at room temperature with 0.1M NaHCO3, 0.5 mg/ml BSA, 0.2% NaN3, pH 8.6.Biotinylated FNfn10 clones were bound in TBST+Ca2+ (TBS 0.1% Tween-20, 2 mM CaCl2).\",\n \"Plates were washed ten times with TBST+Ca2+ and incubated 20 minutes with 2 μg/ml streptavidin-horseradish peroxidase (Oncogene) in TBST+Ca2+ at room temperature.\",\n \"Plates were washed ten times with TBST+Ca2+.\",\n \"Bound peroxidase was detected with ABTS (2,2′-azino-di-3-ethyl-benzthiazoline sulfonic acid) peroxidase substrate (Sigma), 1 mg/ml in 0.1 M NaCitrate, 0.1 M Na2HPO4, pH 4.0 supplemented with 0.03% hydrogen peroxide.\",\n \"Color was allowed to develop for 10-15 minutes and the A400 was read with a SpectraCount ELISA reader (Packard, Downers Grove, Ill.).\",\n \"Flow cytometric detection of cell-binding by biotinylated FNfn10 & FNfn10-3JCLI4: K562 cells (105) transfected with various integrins were resuspended in 5 ml FACS buffer (PBS, 0.5% BSA, 0.1% NaN3), washed, and resuspended in FACS buffer containing either 0.4 μg/ml FNfn10 protein (wild-type of 3JCLI4) or 1 μg/ml LM609, prior to incubation at room temperature for 20 minutes.\",\n \"Cells were then washed with 5 ml FACS buffer, incubated in 1:50 streptavidin-APC (BD Pharmingen, San Diego, Calif.) for 20 minutes and washed again with 5 ml FACS buffer, prior to detection of cell surface fluorescence using a FACSCalibur flow cytometer (Becton Dickinson, San Jose, Calif.).\",\n \"Cell adhesion assays: Fibronectin and vitronectin-coated strips (Chemicon) were rehydrated 15 minutes in PBS.\",\n \"Inhibitor in IMDM/10% FBS was added to the bottom of the well, and 105 cells/well in maintenance media were added to a final volume of 100 μl/well.\",\n \"Cells were incubated 2 hrs at 37° C., 5% CO2, washed three times with PBS at room temperature, and stained 10 minutes with 0.2% crystal violet, 10% ethanol.\",\n \"Cells were then washed three times with PBS and solubilized with a 1:1 ratio of PBS:absolute ethanol for 10 minutes with agitation.\",\n \"A600 was measured using a SpectraCount ELISA reader (Packard).\",\n \"Example 1 Screening of the JCFN-RGD Phage Display Library for αvβ3 Integrin Binding In the FNfn10 structure of SEQ ID No: 2 (wild-type), the FG loop corresponds to residues 78-87, and the RGD sequence to residues 79-81 (residue numbering according to the Protein Data Bank entry 1TTG, Main et al., 1992).\",\n \"Five residues immediately adjacent to the RGD sequence were modified in the FG loop using Kunkel mutagenesis as described above.\",\n \"The JCFN-RGD (XRGDXXXX, SEQ ID No: 4) library contained 1.5×109 independent clones, sufficiently large to include all possible sequences.\",\n \"The library was amplified in 0.0, 0.1, or 1.0 mM IPTG to vary FNfn10 copy number and screened for binding to αvβ3 integrin.\",\n \"After 3 rounds of biopanning, 100-1000 fold increased recovery of phage was observed compared to the initial library.\",\n \"Of 20 phage clones sequenced, 8 contained an XRGDWXEX (SEQ ID No: 26) consensus sequence (FIG.\",\n \"4A).\",\n \"Phage inputs of 109, 108, or 107 pfu were then allowed to bind to αvβ3 or BSA coated wells and bound phages were recovered with 0.2M glycine, pH 2.2.Using phage clones amplified in 1 mM IPTG (high FNfn10 copy number), phage clones containing the consensus were recovered at levels of 0.01-1% input from αvβ3 coated wells; the clones were recovered at approximately 1000-fold lower levels from BSA coated wells and integrin-binding was found to be calcium-dependent.\",\n \"In contrast, clones which lacked the XRGDWXEX (SEQ ID No: 26) motif were recovered at equivalent levels from both αvβ3 coated and BSA coated wells.\",\n \"The RGDWXE-containing clones were amplified in the absence of IPTG so as to decrease the expressed copy number of FNfn10 protein on the phage surface, and the panel of clones were then directly compared in a phage binding assay, for their ability to bind to αvβ3 (FIG.\",\n \"4B).\",\n \"The clones recovered at the highest level were 2JCAV1, 2JCAV2, 3JCAV3, 3JCNI1, and 3JCLI4, all of which were recovered at a level of approximately 0.1% of input phage from αvβ3-coated wells and at 1000-fold lower levels from BSA-coated wells.\",\n \"Therefore, varying FNfn10 copy number on M13 by IPTG concentration did not have an effect on the relative binding efficiency of the clones isolated, suggesting that the binding interaction between the αvβ3 integrin and the phage-displayed FNfn10 derivatives might be of high affinity.\",\n \"The modified FNfn10 proteins expressed by this panel of phage clones were expressed as an isolated protein in E. coli, purified and biotinylated.\",\n \"3JCLI4 (the second-highest recovered clone in phage binding assays), was selected for further study because the purified FNfn10 protein derived from clone 2JCAV1 (the highest recovered clone) proved to be rather insoluble and prone to precipitation.\",\n \"SDS polyacrylamide gel electrophoresis and Coomassie blue staining revealed the presence of a single protein of the expected size (˜15 kDa) within the purified preparation of biotinylated FNfn10-3JCLI4 protein.\",\n \"Western blot analysis using a monoclonal antibody specific for the hexahistidine epitope tag on the purified FNfn10 protein confirmed the identity of this protein, and further analysis revealed that the binding of both the phage clone 3JCLI4 and of the corresponding purified protein (FNfn10-3JCLI4) to αvβ3 integrin was Ca++ dependent and could be eliminated in the presence of EDTA.\",\n \"Example 2 Determining Affinity and Specificity of FNfn10-3JCLI4 for αvβ3 Integrin Biotinylated FNfn10-3JCLI4 and biotinylated wild-type FNfn10 (FNfn10-WT) were allowed to bind to αvβ3, α1β1, or BSA coated wells in approximate half-log increments from 0.0001 to 0.1 μg/ml, corresponding to molar concentrations of 8 pM to 8 nM.\",\n \"Bound biotinylated protein was detected by ELISA (FIG.\",\n \"5A).\",\n \"Assuming a high ratio of free to bound FNfn10-3JCLI4, the equation y=(m1*x/(m2+x)) was fit to the data (R>0.99) to determine the half-maximal binding affinity of 0.01 μg/ml or 800 pM.\",\n \"At each concentration examined, the binding of biotinylated FNfn10-3JCLI4 to αvβ3 was higher than that for FNfn10-WT, and neither protein was found to bind detectably to the α1β1 integrin (i.e., binding to α1β1 was identical to binding to BSA).\",\n \"Next, the kinetics of protein association and dissociation was examined.\",\n \"After allowing biotinylated FNfn10-3JCLI4 to bind to αvβ3-coated wells, 100 fold excess unlabelled FNfn10-3JCLI4 was added to some wells and the plates were incubated for a further 24 hours; control wells received were incubated in the absence of the unlabelled competitor.\",\n \"At the end of this time, the amount of plate-bound biotinylated FNfn10-3JCLI4 was assessed by ELISA assay.\",\n \"No difference was detected between wells that were incubated for 24 hours in the presence or absence of excess unlabelled competitor protein.\",\n \"This suggests that the rate of dissociation of FNfn10-3JCLI4 from αvβ3 is very slow (>24 hours; FIG.\",\n \"5B).\",\n \"To examine the rate of association between αvβ3 and biotinylated 3JCLI4, biotinylated FNfn10-3JCLI4, at concentrations of 0.01, 0.02, or 0.05 μg/ml, was allowed to bind to αvβ3-coated wells for periods of time ranging from one minute to two hours.\",\n \"For all three concentrations, levels of bound FNfn10-3JCLI4 were detectable within one minute and reached saturation levels within two hours (FIG.\",\n \"5C).\",\n \"Therefore, in light of the prolonged dissociation rate revealed in FIG.\",\n \"5B, it can be inferred that FNfn10-3JLCI4 possesses a relatively rapid on-rate for binding to αvβ3.To test the specificity of the interaction between FNfn10-3JCLI4 and αvβ3 integrin, biotinylated FNfn10-3JCLI4 or FNfn10-WT were added to wells coated with equivalent 5 μg/ml concentrations of different purified integrins (FIG.\",\n \"5D).\",\n \"In this assay, the detection of plate-bound FNfn10-3JCLI4 was significant only for those wells which had been coated with αvβ3, and not for wells coated with other integrins, including αvβ5, α5β1, α1β1 and αIIbβ3.Example 3 Determining Relative Affinity of FNfn10-3JCLI4 for αvβ3 Integrin In order to examine the relative affinity of FNfn10-3JCLI4, as compared to other αvβ3 binding molecules, a competition ELISA analysis was performed.\",\n \"In these experiments, biotinylated FNfn10-3JCLI4 at 800 pM was mixed with molar ratios of competitor proteins ranging from 0.01 to 100, and allowed to bind to αvβ3 coated wells (FIGS.\",\n \"6A and 6B).\",\n \"Binding of biotinylated FNfn10-3JCLI4 to αvβ3 was unaffected by the presence of the linear integrin-binding peptide (GRGDSPK, SEQ ID No: 27) or FNfn10-WT, even when these molecules were added at 100-fold molar excess (80 nM), relative to FNfn10-3JCLI4.In contrast, unlabelled FNfn10-3JCLI4 and echistatin inhibited binding of biotinylated FNfn10-3JCLI4 at roughly equal equimolar concentrations (IC50 values were 750 and 930 pM, respectively, as calculated by logarithmic curve-fit).\",\n \"These findings strongly suggest that FNfn10-3JCLI4 and echistatin have roughly equivalent affinities for αvβ3.Echistatin has a published Kd of 330 pM (Kumar et al., 1997).\",\n \"Example 4 Role of FNfn10-3JCLI4 as a Staining Agent for αvβ3 Integrin Biotinylated FNfn10-3JCLI4 (1 μg/ml) was added to wild-type K562 cells and to K562 cells that had been stably transfected with αvβ3 integrin (generous gift of Dr. S. Blystone).\",\n \"Surface bound FNfn10-3JCLI4 was then detected using streptavidin-APC and stained cells were enumerated by flow cytometry.\",\n \"The mean fluorescence intensity of staining on the αvβ3-positive cells was approximately three logs higher than on the non-transfected control cells (FIG.\",\n \"7A), and concentrations of biotinylated FNfn10-3JCLI4 as low as 0.008 μg/ml were found to be capable of effectively and specifically staining K562-αvβ3 cells.\",\n \"Having optimized the staining analysis using the K562-αvβ3 cells, a more complete analysis of the specificity of cell surface binding of biotinylated FNfn10-3JCLI4 was performed, using a panel of K562 cell sublines, each of which had been stably transfected with various integrin heterodimers (αvβ3, αvβ5, αIIbβ3, α4β1, and α4β7; all gifts of S. Blystone) (Blystone et al., 1994).\",\n \"These experiments revealed that FNfn10-3JCLI4 showed a strong selective preference for αvβ3, although it did also react very weakly with αIIbβ3 positive cells.\",\n \"In contrast, FNfn10-WT showed a slight preference for cells expressing αvβ3, but stained all of the K562 lines with low intensity (FIG.\",\n \"7B).\",\n \"One of the unique properties of the FNfn10 scaffold is its high level of physical stability.\",\n \"Therefore, an examination was performed to determine the effect of prolonged incubation at elevated temperatures on the binding activity of FNfn10-3JCLI4, as compared to the αvβ3s-specific monoclonal antibody, LM609.In these studies, FNfn10-3JCLI4 and LM609 were incubated for 24 hours at 4, 20, 37, 50, 65, or 75 degrees Celsius, prior to addition to K562-αvβ3 cells and performance of flow cytometry.\",\n \"As can be observed in FIG.\",\n \"7C, incubation of FNfn10-3JCLI4 at 75° C. for 24 hours had only a modest effect on its cell binding activity; in contrast, the same conditions resulted in the complete elimination of LM609's ability to interact with αvβ3.Example 5 Role of FNfn10-3JCLI4 as an Inhibitor of αvβ3-Dependent Cell Adhesion K562-αvβ3 cells, but not K562 cells, adhere strongly to vitronectin (but not BSA).\",\n \"This property was therefore used as the basis for a quantitative assessment of the ability of FNfn10-3JLCI4 to disrupt αvβ3 dependent cell adhesion.\",\n \"To do this, K562-αvβ3 cells were allowed to adhere to vitronectin-coated wells, and the number of bound cells per well was then quantified by crystal violet staining and cell solubilization.\",\n \"LM609, a commonly used αvβ3 function-blocking antibody, and FNfn10-3JCLI4 were both found to inhibit adhesion of K562-αvβ3 cells to vitronectin, while FNfn10-WT showed little effect in this assay (FIG.\",\n \"8A).\",\n \"A more careful analysis, using a wider range of protein concentrations, revealed that FNfn10-3JCLI4 inhibits αvβ3-dependent adhesion to vitronectin at approximately the same concentrations as the well-characterized αvβ3-binding disintegrin, echistatin (FIG.\",\n \"8B).\",\n \"Based on an exponential curve-fit, echistatin was found to inhibit αvβ3-mediated cell adhesion to vitronectin with an IC50 of 5.9 nM, while FNfn10-3JCLI4 inhibited αvβ3-dependent adhesion to vitronectin with an IC50 of 8.6 nM (R>0.99 in both cases).\",\n \"Finally, the effect of FNfn10-3JCLI4 on αIIbβ3-dependent cell adhesion was also examined, because of the weak binding activity of FNfn10-3JCLI4 for K562-αIIbβ3 cells that was revealed in the flow cytometric analyses (Example 4 above).\",\n \"These experiments revealed that FNfn10-3JCLI4 had no measurable effect on αIIbβ3-mediated cell adhesion to fibronectin, while echistatin inhibited αIIbβ3-dependent adhesion with an IC50 of 10.2 nM (R>0.99; FIG.\",\n \"8C).\",\n \"Discussion of Examples 1-5 In this work, phage display technology and random mutagenesis techniques were applied to carry out affinity maturation of FNfn10 binding to αvβ3 integrin.\",\n \"Using one phage clone that was derived from a biopanning screen against immobilized αvβ3, 3JCLI4, the following was observed: (i) modified FNfn10 phage clones, including 3JCLI4, which bind to αvβ3 integrin all contain an XRGDWXEX (SEQ ID No: 26) consensus and exhibit calcium-dependent binding; (ii) purified FNfn10-3JCLI4 protein binds to immobilized human αvβ3 integrin with much higher affinity than FNfn10-WT and exhibits only background binding to other purified integrins; (iii) binding of FNfn10-3JLCI4 to αvβ3 could be successfully competed using approximately equimolar concentrations of unlabelled FNfn10-3JCLI4 or echistatin, but not by either FNfn10-WT or GRGDSPK peptide (SEQ ID No: 27), even when added at 100-fold molar excess, (iv) FNfn10-3JCLI4 is a sensitive and specific reagent for the detection of human αvβ3 integrin in flow cytometry applications; (v) the binding of FNfn10-3JCLI4 to αvβ3 integrin is maintained even after prolonged exposure of FNfn10-3JCLI4 to high temperature, and (vi) FNfn10-3JCLI4 inhibits cell adhesion via αvβ3 but not αIIbβ3 integrin at nanomolar concentrations.\",\n \"The RGDW sequence that was detected in the αvβ3-selected FNfn10-display phage clones has previously been reported to be preferentially selective for αIIbβ3 integrin over αvβ3, at least in the context of disintegrins such as eristostatin (McLane et al., 1996).\",\n \"RGDW peptides are also used to induce integrin activation and in competition assays with putative αIIbβ3 integrin ligands (Hantgan et al., 1992).\",\n \"Therefore, it was quite unexpected that the consensus sequence (XRGDWXEX, SEQ ID No: 26) bound preferentially to αvβ3 over αIIbβ3 integrins.\",\n \"Because FNfn10-3JCLI4 exhibits no binding to plate-immobilized αIIbβ3 and very low cross-reactivity with αIIbβ3-positive cells in flow cytometric analysis, the tertiary structure of FNfn10 may play a role in the increased specificity of this RGDW-containing molecule for αvβ3 integrin.\",\n \"Alternatively, the glutamic acid residue within the extended consensus motif (SEQ ID No: 26) may be significant.\",\n \"The disintegrins eristostatin and EC6B each contain an aspartic acid residue at the corresponding location (RGDWND, SEQ ID No: 28) (McLane et al., 1996; Scarborough et al., 1993; Marcinkiewicz et al., 2000), and though they both bind αvβ3, they interact preferentially with αIIbβ3.Like the disintegrins, FNfn10-3JCLI4 is a small monomeric molecule with high integrin-binding affinity.\",\n \"In the above Examples, echistatin and FNfn10-3JCLI4 were inferred to have similar binding affinities for αvβ3 because they were both able to compete with biotinylated FNfn10-3JCLI4 for binding to immobilized αvβ3, at roughly equimolar concentrations.\",\n \"Consistent with this, echistatin has a reported Kd of 330 pM for αvβ3 (Kumar et al., 1997), while the value calculated here for the half-maximal binding affinity of FNfn10-3JCLI4 for αvβ3 was 800 pM.\",\n \"Biochemical analyses revealed a rapid rate of association rate between FNfn10-3JCLI4 and αvβ3 (minutes), and a very slow rate of dissociation (>24 hours).\",\n \"These properties, combined with its high affinity for αvβ3, suggested that FNfn10-3JCLI4 might be suitable for use in flow cytometric applications.\",\n \"Experimental analysis revealed this was indeed the case, and these studies also showed that FNfn10-3JCLI4 performed similarly to αvβ3-specific monoclonal antibodies in flow cytometric staining using K562-αvβ3 cells.\",\n \"It should be noted that FNfn10-3JCLI4 is monomeric, unlike monoclonal antibodies (divalent) or other cell-staining reagents used in flow cytometry (MHC:peptide tetramers).\",\n \"Furthermore, FNfn10-3JCLI4 is considerably more stable than a typical monoclonal antibody, as revealed by its ability to continue to bind αvβ3, even after 24 hr.\",\n \"incubation at 75° C. A problem encountered in many molecules that target αvβ3 integrins is their cross-reactivity with other integrin subtypes, particularly other αv or β3 integrins.\",\n \"FNfn10-3JCLI4 was found to bind only to immobilized purified αvβ3, and not to other integrins tested.\",\n \"The failure of FNfn10-WT and FNfn10-3JCLI4 to interact with α5β1 is consistent with previous reports that the ninth and eighth fibronectin type III domains are also required for FNfn10 binding to α5β1 (Altroff et al., 2001).\",\n \"In addition, the inability of FNfn10-3JCLI4 to bind to αvβ5 and αIIbβ3, suggests that its integrin-binding activity is likely dependent on a structural interaction with the αvβ3 complex and not with either subunit alone.\",\n \"The narrow integrin-binding specificity of FNfn10-3JCLI4 can be contrasted with the more broad integrin-binding profile that is exhibited by disintegrins such as eristostatin and kistrin, which bind to both αIIbβ3 and αvβ3 (McLane et al., 1996).\",\n \"Similarly, the penton base protein from adenovirus type 5 binds to both αvβ3 and αvβ5, and even an engineered Fab construct (WOW-1) has been shown to exhibit cross-reactivity with αvβ5 integrin (Pampori et al., 1999).\",\n \"Although FNfn10-3JCLI4 did exhibit a weak interaction with αIIbβ3 in the flow cytometry assay, this will not be a major detriment to the more widespread use of this molecule in cell staining applications because (i) no interaction with αIIbβ3 was detected in biochemical binding studies, (ii) binding to αIIbβ3 positive cells was almost 10-fold weaker than binding to αvβ3-positive cells in the flow cytometric analysis, and (iii) αIIbβ3 expression is limited to platelets.\",\n \"The ability of FNfn10-3JCLI4 to block αvβ3-dependent cell adhesion but not αIIbβ3-mediated cell binding further supports its specificity for αvβ3.The structure of FNfn10-3JCLI4 is advantageous for multiple applications; FNfn10-3JCLI4 lacks disulfide bonds rendering it resistant against reducing agents, is very stable even at high temperature, and can be produced in a bacterial expression system with yields of up to 50-100 mg per liter of culture.\",\n \"As an endogenous human protein, wild-type fibronectin does not elicit an immune response and it is anticipated that FNfn10-3JCLI4 is likely to be similarly non-immunogenic.\",\n \"Furthermore, because FNfn10-3JCLI4 is a monomer that lacks disulfide bonds, it has several advantages over a monoclonal antibody with similar affinity.\",\n \"As a small, single-chain molecule, FNfn10-3JCLI4 could potentially be incorporated into gene delivery vectors (e.g.\",\n \"viral vectors, liposomes) for cell or tissue-specific gene expression.\",\n \"Indeed, the incorporation of low-affinity RGD peptides into various viral vectors, and the addition of bispecific antibody complexes to viral surface components, have already been shown to result in desirable improvements in vector specificity and transduction efficiency (Haisma et al., 2000; Wickham et al., 1996; Wickham et al., 1997; Hart, 1999).\",\n \"αvβ3-positive cells that may be of particular interest for targeted vector systems include dendritic cells (for vaccine applications) as well as osteoclasts and angiogenic vessels.\",\n \"Finally, FNfn10-3JCLI4 will likely have utility as a cancer imaging agent, because of its ability to recognize αvβ3 on tumor vasculature.\",\n \"Example 6 Formation of αvβ3-Directed MRI Diagnostic With Superparamagnetic Iron Oxide Particles Polypeptide monobodies prepared as described in Example 1 will be used to form an MRI diagnostic agent upon conjugation to superparamagnetic iron oxide (SPIO) particles.\",\n \"Dextran-coated SPIO particles can be purchased (Advanced Magnetics, Cambridge, Mass.)\",\n \"or synthesized as described previously (Kresse et al., 1998).\",\n \"SPIO particles are extremely effective relaxation agents, especially as effectors of T2*.\",\n \"The half-life of SPIO particles in blood highly depends on particle size (Moore et al., 2000) and will be an important factor in the design.\",\n \"A long half-life is useful for allowing time for the contrast agent to escape the capillaries and penetrate into tissue.\",\n \"However, αvβ3 is highly accessible to intravascular compounds, and a long blood half-life increases a background, non-specific signal enhancement.\",\n \"Therefore, SPIO particles with diameter >30 nm that have short blood half-lives of 8-20 minutes (Reimer et al., 1992; Moore et al., 2000) will be used.\",\n \"This should allow sufficient time for binding to αvβ3 and an acceptably short time for clearance of unbound agent.\",\n \"Particle size is primarily determined by the amount of dextran used during oxide crystal precipitation.\",\n \"Size distribution will be determined by a dynamic light scattering instrumentation according to known procedures.\",\n \"Conjugation between the polypeptide monobody and SPIO will be achieved using a streptavidin-biotin linkage.\",\n \"Two approaches will be employed.\",\n \"In a first approach, a fusion protein will be expressed from a chimeric gene which includes a polypeptide monobody coding region and streptavidin-biotin coding region linked by an in-frame gene fusion.\",\n \"The chimeric gene will be inserted into an E. coli expression vector (pET derivative) to produce a biotinylated protein.\",\n \"In a second approach, a polypeptide monobody will be expressed containing a free cysteine in the C-terminal flexible tail.\",\n \"Thereafter, the polypeptide monobody will be biotinylated by chemical modification.\",\n \"Unlike antibodies and cyclic peptides, FN3 have no cysteine residues that would complicate cysteine modification.\",\n \"These proteins also contain a poly-histidine tag in the C-terminal tail for purification.\",\n \"The C-terminus of FN3 is located at the opposite side of the protein from the RGD sequence of the FG loop (FIG.\",\n \"1) and, thus, it is expected that these modification will have little, if any, effect on the binding activity of FN3 mutants.\",\n \"Streptavidin will be cross-linked with the dextran coat of the SPIO particles by chemical modification (reductive amination) (Shen et al., 1996).\",\n \"Then, biotinylated FN3 will be attached to this complex, and FN3-biotin-streptavidin-SPIO will be purified using ultrafiltration.\",\n \"REFERENCES Each of the references cited herein or otherwise listed below are expressly incorporated by reference in their entirety into this specification.\",\n \"Adonis et al.\",\n \"(1994) Amer.\",\n \"J. 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\"4,237,224 to Cohen and Boyer Although the invention has been described in detail for the purposes of illustration, it is understood that such detail is solely for that purpose, and variations can be made therein by those skilled in the art without departing from the spirit and scope of the invention which is defined by the following claims.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473770"}}},{"rowIdx":751,"cells":{"text":{"kind":"list like","value":[["Controller of internal combustion engine","Disclosed is an internal combustion engine controller, which is used with a fuel-based torque-on-demand control type, multi-cylinder, direct-injection internal combustion engine to provide an air flow rate control means that excels in response and convergence.","This internal combustion engine controller comprises means for computing a target throttle opening from operating conditions.","The means for computing a target throttle opening includes: a first computing means for determining a target throttle opening by exercising feedback control in accordance with operating conditions including an intake air flow rate; a second computing means for determining a target throttle opening by exercising feed-forward control in accordance with operating conditions; and a third computing means for determining a target throttle opening in accordance with a target throttle opening value determined by the first computing means and a target throttle opening value determined by the second computing means."],["1.An internal combustion engine controller, comprising: means for computing a target throttle opening from operating conditions; wherein the means for computing a target throttle opening comprises: a first computing means for determining a target throttle opening by exercising feedback control in accordance with operating conditions including an intake air flow rate; a second computing means for determining a target throttle opening by exercising feed-forward control in accordance with operating conditions; and a third computing means for determining a target throttle opening in accordance with a target throttle opening value determined by said first computing means and a target throttle opening value determined by said second computing means.","2.An internal combustion engine controller, comprising: a target air flow rate computing means for computing a target air flow rate in accordance with operating conditions; a first computing means for determining a target throttle opening by exercising feedback control in accordance with a difference between said target air flow rate and an actual air flow rate; a second computing means for determining a target throttle opening by exercising feed-forward control in accordance with said target air flow rate; and a third computing means for determining a target throttle opening in accordance with a target throttle opening value determined by said first computing means and a target throttle opening value determined by said second computing means.","3.An internal combustion engine controller, comprising: a control unit for computing a target throttle opening from operating conditions; wherein the control unit comprises: a first computing means for determining a target throttle opening by exercising feedback control in accordance with operating conditions including an intake air flow rate; a second computing means for determining a target throttle opening by exercising feed-forward control in accordance with operating conditions; a third computing means for determining a target throttle opening in accordance with a target throttle opening value determined by said first computing means and a target throttle opening value determined by said second computing means; and a throttle opening control means for driving a throttle in accordance with a target throttle opening calculated by said third computing means.","4.The internal combustion engine controller according to claim 3, wherein said throttle opening control means comprises means for controlling a throttle in such a manner as to reduce a difference between a target throttle opening calculated by said third computing means and a directly or indirectly detected actual throttle opening.","5.The internal combustion engine controller according to claim 1, wherein said target throttle opening computing means comprises a computation permission means for permitting said first target throttle opening computing means to compute a target throttle opening.","6.The internal combustion engine controller according to claim 5, wherein said computation permission means inhibits said first target throttle opening computing means from computing a target throttle opening if means for detecting an actual air flow rate directly or indirectly is found to be abnormal; and wherein said third target throttle opening computing means calculates a target throttle opening in accordance with a second target throttle opening and makes it possible to adjust the actual air flow rate for a target air flow rate.","7.The internal combustion engine controller according to claim 5, wherein said computation permission means inhibits said first target throttle opening computing means from computing a target throttle opening if an accelerator opening is not smaller than a predetermined value or said second target throttle opening is not smaller than a predetermined value; and wherein said third target throttle opening computing means calculates a target throttle opening in accordance with the second target throttle opening and makes it possible to adjust an actual air flow rate to a target air flow rate.","8.The internal combustion engine controller according to claim 5, wherein said computation permission means inhibits said first target throttle opening computing means from calculating a target throttle opening if a difference between a target air flow rate and an actual air flow rate is not greater than a predetermined value; and wherein said third target throttle opening computing means calculates a target throttle opening in accordance with the second target throttle opening and makes it possible to adjust an actual air flow rate to a target air flow rate.","9.The internal combustion engine controller according to claim 1, wherein said target throttle opening computing means comprises a computation permission means for permitting said second target throttle opening computing means to compute a target throttle opening in accordance with operating conditions.","10.The internal combustion engine controller according to claim 9, wherein said computation permission means inhibits said second target throttle opening computing means from computing a target throttle opening if a target exhaust gas re-circulation rate is not lower than a predetermined value; and wherein said third target throttle opening computing means calculates a target throttle opening in accordance with a first target opening and makes it possible to adjust an actual air flow rate to a target air flow rate.","11.The internal combustion engine controller according to claim 1, wherein said target throttle opening computing means makes it possible to adjust an actual air flow rate to a target air flow rate by calculating said third target throttle opening from said first target throttle opening, which is computed in accordance with an actual air flow rate, even if a relationship between said second target throttle opening and the actual air flow rate is significantly changed.","12.The internal combustion engine controller according to claim 2, wherein said target throttle opening computing means comprises a target air flow rate correction means for correcting said target air flow rate; and wherein said first target throttle opening computing means computes a first target throttle opening in accordance with a difference between an actual air flow rate and a target air flow rate calculated by said target air flow rate correction means.","13.The internal combustion engine controller according to claim 12, wherein said target air flow rate correction means represents a response characteristic between said second target throttle opening and an actual air flow rate.","14.The internal combustion engine controller according to claim 1, wherein said second target throttle opening computing means computes a second target throttle opening in accordance with a rotating speed of an internal combustion engine and a target air flow rate.","15.The internal combustion engine controller according to claim 1, further comprising means for computing a target exhaust gas re-circulation rate; wherein said second target throttle opening computing means computes a second target throttle opening in accordance with a rotating speed of an internal combustion engine, a target air flow rate, and the target exhaust gas re-circulation rate.","16.The internal combustion engine controller according to claim 2, wherein said target air flow rate computing means computes said target air flow rate in accordance with a target air-fuel ratio and a target fuel injection quantity.","17.The internal combustion engine controller according to claim 2, further comprising: detection means for detecting exhaust gas components of an internal combustion engine directly or indirectly; and a correction amount computing means for computing an amount of correction for said target air flow rate in accordance with the exhaust gas components detected by said detection means.","18.The internal combustion engine controller according to claim 1, wherein said second target throttle opening computing means computes a second target throttle opening in accordance with an inverse model for a response between an in-cylinder target air flow rate and a throttle opening.","19.The internal combustion engine controller according to claim 1, wherein the actual air flow rate is a value obtained by detecting an in-cylinder air flow rate directly or indirectly."],["<SOH> BACKGROUND OF THE INVENTION <EOH>In recent years, worldwide efforts have been made to provide increased energy savings.","In the field of automotive technology, the development of a fuel-efficient internal combustion engine has been required for energy saving purposes.","The most conspicuous internal combustion engine meeting such a demand is a lean-burn internal combustion engine.","In particular, an in-cylinder fuel injection internal combustion engine, which is a lean-burn internal combustion engine, injects fuel directly into a cylinder to stratify an air-fuel mixture, thereby making it possible to achieve combustion at an air-fuel ratio of higher than 40 and reduce the pump loss.","In a lean-burn in-cylinder fuel injection internal combustion engine system, which is described above, the air flow rate is not proportional to the torque.","Therefore, the lean-burn in-cylinder fuel injection internal combustion engine system generally uses an electronic throttle for electronically controlling the air flow rate unlike a conventional internal combustion engine system.","For the above lean-burn in-cylinder fuel injection internal combustion engine system, torque-on-demand control is required to provide torque desired by the driver at a wide-range air-fuel ratio.","Two types of torque-on-demand control are an air-based type and a fuel-based type.","If air-based torque-on-demand control is exercised, a target torque computation section and a target air-fuel ratio computation section determine a target torque and target air-fuel ratio, respectively, as shown in FIG.","27 .","A target air flow rate computation section for providing the target torque and target air-fuel ratio computes a target air flow rate.","An electronic throttle controls the air flow rate.","An air flow rate sensor detects an actual air flow rate.","A fuel injection quantity computation section determines the quantity of fuel injection from the actual air flow rate and target air-fuel ratio.","If, on the other hand, fuel-based torque-on-demand control is exercised, a target torque computation section determines a target torque as shown in FIG.","28 .","A fuel injection quantity computation section then determines the quantity of fuel injection for providing the target torque.","A target air computation section computes a target air flow rate from the fuel injection quantity and target air-fuel ratio.","An electronic throttle controls the air flow rate.","Further, fuel-based torque-on-demand control can be exercised to provide feedback control over the air flow rate in accordance with a value output by an air flow sensor.","Fuel-based torque-on-demand control described above uses an electronic throttle to exercise air flow rate control after fuel injection quantity determination.","However, a transmission characteristic exists between the electronic throttle and cylinder.","More specifically, a transient phenomenon occurs because it generally takes tens to hundreds of milliseconds for the air flow rate controlled near the electronic throttle to arrive in a cylinder as shown in FIG.","29 .","In an in-cylinder fuel injection internal combustion engine, on the other hand, fuel injection directly occurs within a cylinder.","Therefore, the transmission characteristic of the fuel injection side is smaller than that of the air side because time is merely wasted by intermittent combustion.","Meanwhile, the exhaust pipe for an internal combustion engine is usually provided with a three-way catalyst or a catalyst having a three-way catalytic function as an exhaust gas emission purification system.","As shown in FIG.","30 , the three-way catalyst efficiently purifies carbon hydride (HC), carbon monoxide (CO), which are reducers, and nitrogen oxide (NOx), which is an oxidant, only in the neighborhood of a theoretical air-fuel ratio.","From the viewpoint of exhaust gas emission reduction, it is desirable that the air-fuel ratio for an internal combustion engine be adjusted for the theoretical air-fuel ratio.","As regards a lean air-fuel ratio, there is a correlation between the air fuel ratio and internal combustion engine combustion stability as shown in FIG.","31 .","It is therefore necessary to control the air fuel ratio for the purpose of providing combustion stability of an internal combustion engine.","Thus, when an air flow rate transient phenomenon in a cylinder is considered, the fuel injection quantity needs to be controlled from the viewpoint of exhaust gas emission reduction for theoretical air-fuel ratio or from the viewpoint of internal combustion engine combustion stability for lean air-fuel ratio.","Further, the torque of an internal combustion engine is determined almost conclusively by the fuel injection quantity.","Therefore, the torque-response is determined by the transient characteristic of air flow rate.","As is obvious from the above description, the most important tasks to be accomplished for an internal combustion engine that provides fuel-based torque-on-demand control are to improve the response to air flow rate control and convergence performance, minimize the variations among mass-produced products, and improve the robustness for changes with time.","A technology disclosed by JP-A No.","2000-97086 provides control over the air-fuel ratio of an internal combustion engine.","When the target air flow rate changes, this technology provides delay compensation to improve the air flow rate response within a cylinder by temporarily permitting the throttle opening to overshoot the degree of throttle opening for achieving the target air flow rate.","However, this control method cannot exercise air flow rate control if the air flow sensor is faulty because it computes the target throttle opening in accordance with the deviation between the actual air flow rate and target air flow rate.","Another technology disclosed by JP-A No.","6-146950 changes the throttle opening by a predetermined amount, if there is any change in the target air-fuel ratio, to eliminate any inadequate feedback response portion of the actual air flow rate with a view toward response improvement.","However, since this control method does not detect the actual air flow rate, it cannot properly respond, for instance, to air density changes at a high altitude and exhibits low robustness for air flow rate control accuracy in relation to various environmental changes such as throttle control sensor and actuator characteristic variations.","In consideration of the problems described above, it is an object of the present invention to provide an internal combustion engine controller that is capable of exercising high-performance air flow rate control of a fuel-based torque-on-demand control type, in-cylinder fuel injection internal combustion engine while providing improved response and convergence and enhanced robustness."],["<SOH> SUMMARY OF THE INVENTION <EOH>To achieve the above object, an internal combustion engine controller of the present invention basically comprises means for computing a target throttle opening in accordance with operating conditions.","The means for computing a target throttle opening comprises a first computing means, which determines a target throttle opening by exercising feedback control in accordance with operating conditions including an intake air volume; a second computing means, which determines a target throttle opening by exercising feed-forward control in accordance with the operating conditions; and a third computing means, which determines a target throttle opening in accordance with the respective target throttle opening values determined by the first computing means and second computing means.","In another aspect of the present invention, an internal combustion engine controller comprises: a target air flow rate computing means for computing a target air flow rate in accordance with operating conditions; a first computing means for determining a target throttle opening by exercising feedback control in accordance with a difference between the above target air flow rate and an actual air flow rate; a second computing means for determining a target throttle opening by exercising feed-forward control in accordance with the above target air flow rate; and a third computing means for determining a target throttle opening in accordance with the respective target throttle opening values determined by the first computing means and second computing means.","In still another aspect of the present invention, an internal combustion engine controller for computing a target throttle opening in accordance with operating conditions comprises: a first computing means for determining a target throttle opening by exercising feedback control in accordance with operating conditions including the intake air volume; a second computing means for determining a target throttle opening by exercising feed-forward control in accordance with the operating conditions; a third computing means for determining a target throttle opening in accordance with the target throttle opening values determined by the first computing means and second computing means; and a throttle opening control means for driving a throttle valve in accordance with the target throttle calculated by the third computing means.","As indicated by the configuration described above, the internal combustion engine controller of the present invention comprises a first throttle opening computing means for computing a throttle opening by exercising feedback control (F/B control) in accordance with the actual air flow rate, target air flow rate, and other operating conditions and a second throttle opening computing means for computing a throttle opening by exercising feed-forward control (F/F control) in accordance with the target air flow rate and other operating conditions.","Therefore, the internal combustion engine controller of the present invention can improve the air flow rate calculation response and convergence, reduce the exhaust gas emission, and provides improved torque response without being affected by various environmental changes or sensor, actuator, and other component variations.","In other words, the internal combustion engine controller of the present invention is particularly suitable for a fuel-based torque-on-demand control type, internal combustion engine.","It is basically characterized by comprising a second throttle opening computing means for computing a target throttle opening from a target air flow rate and a first throttle opening computing means for computing a target throttle opening from the deviation between a target air flow rate and actual air flow rate.","Common feedback control computes a manipulated variable in accordance with the resulting controlled variable.","It is therefore said that the response of feedback control is theoretically inferior to that of feed-forward control.","On the other hand, it is said that feed-forward control excels in response but exhibits low robustness against disturbance.","The disadvantages of these two control functions are offset by the internal combustion engine controller of the present invention.","Concisely, the internal combustion engine controller of the present invention provides a target throttle opening computing section that is capable of exercising feed-forward control, which is highly responsive, and feedback control, which is robust against disturbance.","Consequently, the internal combustion engine controller of the present invention provides not only improved response but also enhanced robustness against disturbance particularly in air flow rate control over a fuel-based torque-on-demand control type, internal combustion engine.","To achieve another object, the internal combustion engine controller of the present invention uses the throttle opening control means, which comprises means for controlling the throttle valve in such a manner as to reduce the difference between a directly or indirectly detected actual throttle opening value and a target throttle opening value calculated by the third computing means.","As indicated by the configuration described above, the internal combustion engine controller of the present invention provides feedback control, which computes the manipulated variable of the throttle in accordance with the difference between the actual throttle opening and a target throttle opening calculated by the third target throttle opening computing means.","Therefore, the manipulated variable for throttle drive can be computed from the target throttle opening and actual throttle opening.","It means that the manipulated variable for throttle drive indicates a duty ratio that is entered into a drive circuit for controlling a throttle motor drive current.","Thus, the manipulated variable for throttle drive can be accurately calculated by means of PID control.","In addition to feedback control, the internal combustion engine controller of the present invention also provides feed-forward control, which computes the manipulated variable of the throttle in accordance with only the target throttle opening calculated by the third target throttle opening computing means.","Therefore, computation control can be exercised with high responsiveness.","The feed-forward control portion may be a gain only.","In such an instance, the manipulated variable for throttle opening control can be computed directly from a target air flow rate without computing a target throttle opening.","To achieve another object, the internal combustion engine controller of the present invention uses the target throttle opening computing means, which comprises a computation permission means for permitting the first target throttle opening computing means to compute a target throttle opening.","The computation permission means inhibits the first target throttle opening computing means from computing a target throttle opening if the means for detecting the actual air flow rate directly or indirectly is found to be abnormal.","The third target throttle opening computing means calculates a target throttle opening in accordance with the second target throttle opening so that the actual air flow rate can be adjusted for the target air flow rate.","As indicated in the configuration described above, the internal combustion engine controller of the present invention comprises a computation permission means for permitting the first target throttle opening computing means to compute a target throttle opening.","Therefore, the first target throttle opening computing means can be permitted to compute a target throttle opening when, for instance, the opening of an internal combustion engine accelerator is normal, the air flow sensor function is normal, the throttle is normal, or the elapsed time from startup is as specified.","It means that target throttle opening computations can be performed in response to changes in the internal combustion engine environment.","Even when the means for detecting the air flow rate directly or indirectly is found to be abnormal, it is possible to inhibit the first target throttle opening computing means from computing the first target throttle opening, cause the second target throttle opening computing means to calculate the second target throttle opening as the third target throttle opening, and control the actual air flow rate in accordance with the third target throttle opening.","That is, if the means for detecting the air flow rate of the air flow sensor or the like is faulty, the actual air flow rate can be controlled by inhibiting the target throttle opening from being computed by feedback control and using the target throttle opening computed by feed-forward control as the target opening.","Therefore, the air flow rate can be preferably controlled even when the air flow sensor or the like is faulty.","To achieve another object, the internal combustion engine controller of the present invention uses the computation permission means to inhibit the first target throttle opening computing means from computing the target throttle opening if the accelerator opening is not smaller than a predetermined value or the second target throttle opening is not smaller than a predetermined value.","The third target throttle opening computing means computes the target throttle opening in accordance with the second target throttle opening and makes it possible to adjust the actual air flow rate for the target air flow rate.","The computation permission means inhibits the first target throttle opening computing means from computing the target throttle opening if the difference between the target air flow rate and actual air flow rate is not greater than a predetermined value.","The third target throttle opening computing means calculates the target throttle opening in accordance with the second target throttle opening and makes it possible to adjust the actual air flow rate to the target air flow rate.","To achieve another object, the internal combustion engine controller of the present invention uses the target throttle opening computing means, which comprises a computation permission means for permitting the second target throttle opening computing means to compute the target throttle opening in accordance with the operating conditions.","The computation permission means inhibits the second target throttle opening computing means from computing the target throttle opening if a target exhaust gas re-circulation rate is not smaller than a predetermined value.","The third target throttle opening computing means calculates the target throttle opening in accordance with the first target opening and makes it possible to adjust the actual air flow rate to the target air flow rate.","As indicated in the configuration described above, the internal combustion engine controller of the present invention comprises a second computation permission means, which permits the second target throttle opening computing means to compute a throttle opening.","Therefore, if, for instance, the EGR rate is as specified or the air flow sensor is normal, the second target throttle opening computing means can be permitted to computer the target throttle opening.","Thus, it is possible to compute the target throttle opening that is responsive to changes in the internal combustion engine environment.","To achieve another object, the internal combustion engine controller of the present invention uses the target throttle opening computing means to calculate the third target throttle opening from the first target throttle opening, which is computed in accordance with the actual air flow rate even when the relationship between the second target throttle opening and actual air flow rate significantly changes, and makes it possible to adjust the actual air flow rate for the target air flow rate.","As indicated in the configuration described above, the internal combustion engine controller of the present invention exercises feedback control to provide computation control so as to adjust the actual air flow rate for the target air flow rate and calculate the target throttle opening even when the relationship between the target throttle opening computed by feed-forward control and the actual air flow rate significantly changes due to an air density change, foreign matter inclusion in an intake pipe, or other environmental change.","Therefore, the air flow rate can be accurately controlled in response to the above environmental changes.","To achieve another object, the internal combustion engine controller of the present invention uses the target throttle opening computing means, which comprises a target air flow rate correction means for correcting the target air flow rate.","The first target throttle opening computing means computes the first target throttle opening in accordance with the difference between the target air flow rate calculated by the target air flow rate correction means and the actual air flow rate.","The target air flow rate correction means represents a response characteristic between the second target throttle opening and actual air flow rate.","As indicated by the configuration described above, the internal combustion engine controller of the present invention comprises the target air flow rate correction means.","Therefore, it can accurately compute a target air flow rate while improving the transmission characteristic by compensating for a time delay involved in intake air flow from the throttle to a cylinder.","To achieve another object, the internal combustion engine controller of the present invention uses the second target throttle opening computing means to compute the second target throttle opening in accordance with the internal combustion engine's rotating speed and the target air flow rate.","As indicated by the configuration described above, the internal combustion engine controller of the present invention can exercise feed-forward control to compute the second target throttle opening, for instance, from a map depicting the relationship between the internal combustion engine's rotating speed and the target air flow rate.","To achieve another object, the internal combustion engine controller of the present invention comprises means for computing the target exhaust gas re-circulation rate.","Further, the second target throttle opening computing means computes the second target throttle opening in accordance with the internal combustion engine's rotating speed, the target air flow rate, and the target exhaust gas re-circulation rate.","To achieve another object, the internal combustion engine controller of the present invention uses the target air flow rate computing means to compute the target air flow rate in accordance with the target air-fuel ratio and target fuel injection quantity.","As indicated by the configuration described above, the internal combustion engine controller of the present invention can correct the second target throttle opening in accordance with the exhaust gas re-circulation rate when the EGR gas recirculates.","More specifically, it can determine the target EGR rate from, for instance, a map depicting the relationship between a target combustion pressure torque and internal combustion engine's rotating speed, determine a throttle opening correction factor from, for instance, a map depicting the relationship between the target EGR rate and target air flow rate, and correct the second target throttle opening.","To achieve another object, the internal combustion engine controller of the present invention comprises a detection means for detecting the internal combustion engine's exhaust gas components directly or indirectly, and a correction amount computing means for computing the correction amount for the target air flow rate in accordance with the exhaust gas components detected by the detection means.","As indicated by the configuration described above, the internal combustion engine controller of the present invention corrects the target air flow rate with a correction amount (manipulated variable) that is used to compute (by means of air-fuel ratio feedback control) the correction amount for the target air flow rate in accordance with a signal from an A/F sensor or other exhaust gas component detection means.","It is therefore possible to calculate a target air flow rate with increased accuracy and compute a target throttle opening.","To achieve another object, the internal combustion engine controller of the present invention uses the second target throttle opening computing means to compute the second target throttle opening in accordance with an inverse model for the response between an in-cylinder target air flow rate and the throttle.","To achieve still another object, the internal combustion engine controller of the present invention uses an in-cylinder air flow rate value, which is detected directly or indirectly, as an actual air flow rate value."],["FIELD OF THE INVENTION The present invention related to internal combustion engine controllers, and more particularly to an internal combustion engine controller for accurately adjusting the air flow rate by controlling the air flow rate with an electronic control valve.","BACKGROUND OF THE INVENTION In recent years, worldwide efforts have been made to provide increased energy savings.","In the field of automotive technology, the development of a fuel-efficient internal combustion engine has been required for energy saving purposes.","The most conspicuous internal combustion engine meeting such a demand is a lean-burn internal combustion engine.","In particular, an in-cylinder fuel injection internal combustion engine, which is a lean-burn internal combustion engine, injects fuel directly into a cylinder to stratify an air-fuel mixture, thereby making it possible to achieve combustion at an air-fuel ratio of higher than 40 and reduce the pump loss.","In a lean-burn in-cylinder fuel injection internal combustion engine system, which is described above, the air flow rate is not proportional to the torque.","Therefore, the lean-burn in-cylinder fuel injection internal combustion engine system generally uses an electronic throttle for electronically controlling the air flow rate unlike a conventional internal combustion engine system.","For the above lean-burn in-cylinder fuel injection internal combustion engine system, torque-on-demand control is required to provide torque desired by the driver at a wide-range air-fuel ratio.","Two types of torque-on-demand control are an air-based type and a fuel-based type.","If air-based torque-on-demand control is exercised, a target torque computation section and a target air-fuel ratio computation section determine a target torque and target air-fuel ratio, respectively, as shown in FIG.","27.A target air flow rate computation section for providing the target torque and target air-fuel ratio computes a target air flow rate.","An electronic throttle controls the air flow rate.","An air flow rate sensor detects an actual air flow rate.","A fuel injection quantity computation section determines the quantity of fuel injection from the actual air flow rate and target air-fuel ratio.","If, on the other hand, fuel-based torque-on-demand control is exercised, a target torque computation section determines a target torque as shown in FIG.","28.A fuel injection quantity computation section then determines the quantity of fuel injection for providing the target torque.","A target air computation section computes a target air flow rate from the fuel injection quantity and target air-fuel ratio.","An electronic throttle controls the air flow rate.","Further, fuel-based torque-on-demand control can be exercised to provide feedback control over the air flow rate in accordance with a value output by an air flow sensor.","Fuel-based torque-on-demand control described above uses an electronic throttle to exercise air flow rate control after fuel injection quantity determination.","However, a transmission characteristic exists between the electronic throttle and cylinder.","More specifically, a transient phenomenon occurs because it generally takes tens to hundreds of milliseconds for the air flow rate controlled near the electronic throttle to arrive in a cylinder as shown in FIG.","29.In an in-cylinder fuel injection internal combustion engine, on the other hand, fuel injection directly occurs within a cylinder.","Therefore, the transmission characteristic of the fuel injection side is smaller than that of the air side because time is merely wasted by intermittent combustion.","Meanwhile, the exhaust pipe for an internal combustion engine is usually provided with a three-way catalyst or a catalyst having a three-way catalytic function as an exhaust gas emission purification system.","As shown in FIG.","30, the three-way catalyst efficiently purifies carbon hydride (HC), carbon monoxide (CO), which are reducers, and nitrogen oxide (NOx), which is an oxidant, only in the neighborhood of a theoretical air-fuel ratio.","From the viewpoint of exhaust gas emission reduction, it is desirable that the air-fuel ratio for an internal combustion engine be adjusted for the theoretical air-fuel ratio.","As regards a lean air-fuel ratio, there is a correlation between the air fuel ratio and internal combustion engine combustion stability as shown in FIG.","31.It is therefore necessary to control the air fuel ratio for the purpose of providing combustion stability of an internal combustion engine.","Thus, when an air flow rate transient phenomenon in a cylinder is considered, the fuel injection quantity needs to be controlled from the viewpoint of exhaust gas emission reduction for theoretical air-fuel ratio or from the viewpoint of internal combustion engine combustion stability for lean air-fuel ratio.","Further, the torque of an internal combustion engine is determined almost conclusively by the fuel injection quantity.","Therefore, the torque-response is determined by the transient characteristic of air flow rate.","As is obvious from the above description, the most important tasks to be accomplished for an internal combustion engine that provides fuel-based torque-on-demand control are to improve the response to air flow rate control and convergence performance, minimize the variations among mass-produced products, and improve the robustness for changes with time.","A technology disclosed by JP-A No.","2000-97086 provides control over the air-fuel ratio of an internal combustion engine.","When the target air flow rate changes, this technology provides delay compensation to improve the air flow rate response within a cylinder by temporarily permitting the throttle opening to overshoot the degree of throttle opening for achieving the target air flow rate.","However, this control method cannot exercise air flow rate control if the air flow sensor is faulty because it computes the target throttle opening in accordance with the deviation between the actual air flow rate and target air flow rate.","Another technology disclosed by JP-A No.","6-146950 changes the throttle opening by a predetermined amount, if there is any change in the target air-fuel ratio, to eliminate any inadequate feedback response portion of the actual air flow rate with a view toward response improvement.","However, since this control method does not detect the actual air flow rate, it cannot properly respond, for instance, to air density changes at a high altitude and exhibits low robustness for air flow rate control accuracy in relation to various environmental changes such as throttle control sensor and actuator characteristic variations.","In consideration of the problems described above, it is an object of the present invention to provide an internal combustion engine controller that is capable of exercising high-performance air flow rate control of a fuel-based torque-on-demand control type, in-cylinder fuel injection internal combustion engine while providing improved response and convergence and enhanced robustness.","SUMMARY OF THE INVENTION To achieve the above object, an internal combustion engine controller of the present invention basically comprises means for computing a target throttle opening in accordance with operating conditions.","The means for computing a target throttle opening comprises a first computing means, which determines a target throttle opening by exercising feedback control in accordance with operating conditions including an intake air volume; a second computing means, which determines a target throttle opening by exercising feed-forward control in accordance with the operating conditions; and a third computing means, which determines a target throttle opening in accordance with the respective target throttle opening values determined by the first computing means and second computing means.","In another aspect of the present invention, an internal combustion engine controller comprises: a target air flow rate computing means for computing a target air flow rate in accordance with operating conditions; a first computing means for determining a target throttle opening by exercising feedback control in accordance with a difference between the above target air flow rate and an actual air flow rate; a second computing means for determining a target throttle opening by exercising feed-forward control in accordance with the above target air flow rate; and a third computing means for determining a target throttle opening in accordance with the respective target throttle opening values determined by the first computing means and second computing means.","In still another aspect of the present invention, an internal combustion engine controller for computing a target throttle opening in accordance with operating conditions comprises: a first computing means for determining a target throttle opening by exercising feedback control in accordance with operating conditions including the intake air volume; a second computing means for determining a target throttle opening by exercising feed-forward control in accordance with the operating conditions; a third computing means for determining a target throttle opening in accordance with the target throttle opening values determined by the first computing means and second computing means; and a throttle opening control means for driving a throttle valve in accordance with the target throttle calculated by the third computing means.","As indicated by the configuration described above, the internal combustion engine controller of the present invention comprises a first throttle opening computing means for computing a throttle opening by exercising feedback control (F/B control) in accordance with the actual air flow rate, target air flow rate, and other operating conditions and a second throttle opening computing means for computing a throttle opening by exercising feed-forward control (F/F control) in accordance with the target air flow rate and other operating conditions.","Therefore, the internal combustion engine controller of the present invention can improve the air flow rate calculation response and convergence, reduce the exhaust gas emission, and provides improved torque response without being affected by various environmental changes or sensor, actuator, and other component variations.","In other words, the internal combustion engine controller of the present invention is particularly suitable for a fuel-based torque-on-demand control type, internal combustion engine.","It is basically characterized by comprising a second throttle opening computing means for computing a target throttle opening from a target air flow rate and a first throttle opening computing means for computing a target throttle opening from the deviation between a target air flow rate and actual air flow rate.","Common feedback control computes a manipulated variable in accordance with the resulting controlled variable.","It is therefore said that the response of feedback control is theoretically inferior to that of feed-forward control.","On the other hand, it is said that feed-forward control excels in response but exhibits low robustness against disturbance.","The disadvantages of these two control functions are offset by the internal combustion engine controller of the present invention.","Concisely, the internal combustion engine controller of the present invention provides a target throttle opening computing section that is capable of exercising feed-forward control, which is highly responsive, and feedback control, which is robust against disturbance.","Consequently, the internal combustion engine controller of the present invention provides not only improved response but also enhanced robustness against disturbance particularly in air flow rate control over a fuel-based torque-on-demand control type, internal combustion engine.","To achieve another object, the internal combustion engine controller of the present invention uses the throttle opening control means, which comprises means for controlling the throttle valve in such a manner as to reduce the difference between a directly or indirectly detected actual throttle opening value and a target throttle opening value calculated by the third computing means.","As indicated by the configuration described above, the internal combustion engine controller of the present invention provides feedback control, which computes the manipulated variable of the throttle in accordance with the difference between the actual throttle opening and a target throttle opening calculated by the third target throttle opening computing means.","Therefore, the manipulated variable for throttle drive can be computed from the target throttle opening and actual throttle opening.","It means that the manipulated variable for throttle drive indicates a duty ratio that is entered into a drive circuit for controlling a throttle motor drive current.","Thus, the manipulated variable for throttle drive can be accurately calculated by means of PID control.","In addition to feedback control, the internal combustion engine controller of the present invention also provides feed-forward control, which computes the manipulated variable of the throttle in accordance with only the target throttle opening calculated by the third target throttle opening computing means.","Therefore, computation control can be exercised with high responsiveness.","The feed-forward control portion may be a gain only.","In such an instance, the manipulated variable for throttle opening control can be computed directly from a target air flow rate without computing a target throttle opening.","To achieve another object, the internal combustion engine controller of the present invention uses the target throttle opening computing means, which comprises a computation permission means for permitting the first target throttle opening computing means to compute a target throttle opening.","The computation permission means inhibits the first target throttle opening computing means from computing a target throttle opening if the means for detecting the actual air flow rate directly or indirectly is found to be abnormal.","The third target throttle opening computing means calculates a target throttle opening in accordance with the second target throttle opening so that the actual air flow rate can be adjusted for the target air flow rate.","As indicated in the configuration described above, the internal combustion engine controller of the present invention comprises a computation permission means for permitting the first target throttle opening computing means to compute a target throttle opening.","Therefore, the first target throttle opening computing means can be permitted to compute a target throttle opening when, for instance, the opening of an internal combustion engine accelerator is normal, the air flow sensor function is normal, the throttle is normal, or the elapsed time from startup is as specified.","It means that target throttle opening computations can be performed in response to changes in the internal combustion engine environment.","Even when the means for detecting the air flow rate directly or indirectly is found to be abnormal, it is possible to inhibit the first target throttle opening computing means from computing the first target throttle opening, cause the second target throttle opening computing means to calculate the second target throttle opening as the third target throttle opening, and control the actual air flow rate in accordance with the third target throttle opening.","That is, if the means for detecting the air flow rate of the air flow sensor or the like is faulty, the actual air flow rate can be controlled by inhibiting the target throttle opening from being computed by feedback control and using the target throttle opening computed by feed-forward control as the target opening.","Therefore, the air flow rate can be preferably controlled even when the air flow sensor or the like is faulty.","To achieve another object, the internal combustion engine controller of the present invention uses the computation permission means to inhibit the first target throttle opening computing means from computing the target throttle opening if the accelerator opening is not smaller than a predetermined value or the second target throttle opening is not smaller than a predetermined value.","The third target throttle opening computing means computes the target throttle opening in accordance with the second target throttle opening and makes it possible to adjust the actual air flow rate for the target air flow rate.","The computation permission means inhibits the first target throttle opening computing means from computing the target throttle opening if the difference between the target air flow rate and actual air flow rate is not greater than a predetermined value.","The third target throttle opening computing means calculates the target throttle opening in accordance with the second target throttle opening and makes it possible to adjust the actual air flow rate to the target air flow rate.","To achieve another object, the internal combustion engine controller of the present invention uses the target throttle opening computing means, which comprises a computation permission means for permitting the second target throttle opening computing means to compute the target throttle opening in accordance with the operating conditions.","The computation permission means inhibits the second target throttle opening computing means from computing the target throttle opening if a target exhaust gas re-circulation rate is not smaller than a predetermined value.","The third target throttle opening computing means calculates the target throttle opening in accordance with the first target opening and makes it possible to adjust the actual air flow rate to the target air flow rate.","As indicated in the configuration described above, the internal combustion engine controller of the present invention comprises a second computation permission means, which permits the second target throttle opening computing means to compute a throttle opening.","Therefore, if, for instance, the EGR rate is as specified or the air flow sensor is normal, the second target throttle opening computing means can be permitted to computer the target throttle opening.","Thus, it is possible to compute the target throttle opening that is responsive to changes in the internal combustion engine environment.","To achieve another object, the internal combustion engine controller of the present invention uses the target throttle opening computing means to calculate the third target throttle opening from the first target throttle opening, which is computed in accordance with the actual air flow rate even when the relationship between the second target throttle opening and actual air flow rate significantly changes, and makes it possible to adjust the actual air flow rate for the target air flow rate.","As indicated in the configuration described above, the internal combustion engine controller of the present invention exercises feedback control to provide computation control so as to adjust the actual air flow rate for the target air flow rate and calculate the target throttle opening even when the relationship between the target throttle opening computed by feed-forward control and the actual air flow rate significantly changes due to an air density change, foreign matter inclusion in an intake pipe, or other environmental change.","Therefore, the air flow rate can be accurately controlled in response to the above environmental changes.","To achieve another object, the internal combustion engine controller of the present invention uses the target throttle opening computing means, which comprises a target air flow rate correction means for correcting the target air flow rate.","The first target throttle opening computing means computes the first target throttle opening in accordance with the difference between the target air flow rate calculated by the target air flow rate correction means and the actual air flow rate.","The target air flow rate correction means represents a response characteristic between the second target throttle opening and actual air flow rate.","As indicated by the configuration described above, the internal combustion engine controller of the present invention comprises the target air flow rate correction means.","Therefore, it can accurately compute a target air flow rate while improving the transmission characteristic by compensating for a time delay involved in intake air flow from the throttle to a cylinder.","To achieve another object, the internal combustion engine controller of the present invention uses the second target throttle opening computing means to compute the second target throttle opening in accordance with the internal combustion engine's rotating speed and the target air flow rate.","As indicated by the configuration described above, the internal combustion engine controller of the present invention can exercise feed-forward control to compute the second target throttle opening, for instance, from a map depicting the relationship between the internal combustion engine's rotating speed and the target air flow rate.","To achieve another object, the internal combustion engine controller of the present invention comprises means for computing the target exhaust gas re-circulation rate.","Further, the second target throttle opening computing means computes the second target throttle opening in accordance with the internal combustion engine's rotating speed, the target air flow rate, and the target exhaust gas re-circulation rate.","To achieve another object, the internal combustion engine controller of the present invention uses the target air flow rate computing means to compute the target air flow rate in accordance with the target air-fuel ratio and target fuel injection quantity.","As indicated by the configuration described above, the internal combustion engine controller of the present invention can correct the second target throttle opening in accordance with the exhaust gas re-circulation rate when the EGR gas recirculates.","More specifically, it can determine the target EGR rate from, for instance, a map depicting the relationship between a target combustion pressure torque and internal combustion engine's rotating speed, determine a throttle opening correction factor from, for instance, a map depicting the relationship between the target EGR rate and target air flow rate, and correct the second target throttle opening.","To achieve another object, the internal combustion engine controller of the present invention comprises a detection means for detecting the internal combustion engine's exhaust gas components directly or indirectly, and a correction amount computing means for computing the correction amount for the target air flow rate in accordance with the exhaust gas components detected by the detection means.","As indicated by the configuration described above, the internal combustion engine controller of the present invention corrects the target air flow rate with a correction amount (manipulated variable) that is used to compute (by means of air-fuel ratio feedback control) the correction amount for the target air flow rate in accordance with a signal from an A/F sensor or other exhaust gas component detection means.","It is therefore possible to calculate a target air flow rate with increased accuracy and compute a target throttle opening.","To achieve another object, the internal combustion engine controller of the present invention uses the second target throttle opening computing means to compute the second target throttle opening in accordance with an inverse model for the response between an in-cylinder target air flow rate and the throttle.","To achieve still another object, the internal combustion engine controller of the present invention uses an in-cylinder air flow rate value, which is detected directly or indirectly, as an actual air flow rate value.","BRIEF DESCRIPTION OF THE DRAWINGS FIG.","1 illustrates the overall configuration of an internal combustion engine that comprises an internal combustion engine controller according to the present invention; FIG.","2 illustrates the internal configuration of an internal combustion engine controller shown in FIG.","1; FIG.","3 is a control block diagram that illustrates a first embodiment of an internal combustion engine controller according to the present invention; FIG.","4 is a block diagram that illustrates a target torque computing section shown in the control block diagram in FIG.","3; FIG.","5 is a block diagram that illustrates a fuel injection quantity computing section shown in the control block diagram in FIG.","3; FIG.","6 is a block diagram that illustrates a fuel injection quantity correction section shown in the control block diagram in FIG.","3; FIG.","7 is a block diagram that illustrates a target equivalence ratio computing section shown in the control block diagram in FIG.","3; FIG.","8 is a block diagram that illustrates a target air flow rate computing section shown in the control block diagram in FIG.","3; FIG.","9 is a block diagram that illustrates an actual air flow rate computing section shown in the control block diagram in FIG.","3; FIG.","10 is a block diagram that illustrates a target throttle opening computing section shown in the control block diagram in FIG.","3; FIG.","11 is a block diagram that illustrates a throttle opening control section for the target throttle opening computing section shown in FIG.","10; FIG.","12 is a control block diagram that illustrates an internal combustion engine controller according to a second embodiment of the present invention; FIG.","13 is a control block diagram that illustrates a throttle opening computing section shown in the control block diagram in FIG.","12; FIG.","14 is a control block diagram that illustrates a target throttle opening computing section of an internal combustion engine controller according to a third embodiment of the present invention; FIG.","15 is a control block diagram that illustrates a feedback control target value filter for the target throttle opening computing section shown in FIG.","14; FIG.","16 is a control block diagram that illustrates a target throttle opening computing section of an internal combustion engine controller according to a fourth embodiment of the present invention; FIG.","17 is a block diagram that illustrates a feedback control permission judgment section for the target throttle opening computing section shown in FIG.","16; FIG.","18 is a control block diagram that illustrates a target throttle opening computing section of an internal combustion engine controller according to a fifth embodiment of the present invention; FIG.","19 is a control block diagram that illustrates a feed-forward control permission judgment section for the target throttle opening computing section shown in FIG.","18; FIG.","20 is a control block diagram that illustrates a target throttle opening computing section of an internal combustion engine controller according to a sixth embodiment of the present invention; FIG.","21 is a control block diagram that illustrates an actual air flow rate computing section of an internal combustion engine controller according to a seventh embodiment of the present invention; FIG.","22 is a control block diagram that illustrates a target throttle opening computing section of an internal combustion engine controller according to a seventh embodiment of the present invention; FIG.","23 is a control block diagram illustrating an inverse model for feed-forward control, which is within the target throttle opening computing section shown in FIG.","22; FIG.","24 is a control block diagram that illustrates an internal combustion engine controller according to an eighth embodiment of the present invention; FIG.","25 is a control block diagram that illustrates an air-fuel ratio feedback control section shown in FIG.","24; FIG.","26 is a control block diagram that illustrates a target air flow rate computing section shown in FIG.","24; FIG.","27 is a block diagram that illustrates a conventional air-based torque-on-demand control; FIG.","28 is a block diagram that illustrates a conventional fuel-based torque-on-demand control; FIG.","29 shows in-cylinder air flow rate transient characteristic curves that are prevalent when an internal combustion engine's target air flow rate changes; FIG.","30 shows HC, CO, and NOx purification characteristic curves of an internal combustion engine's three-way catalyst; and FIG.","30 shows a correlation between an internal combustion engine's air-fuel ratio and internal combustion engine stability.","DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Embodiments of an internal combustion engine controller according to the present invention will now be described in detail with reference to the accompanying drawings.","FIG.","1 illustrates an overall internal combustion engine system that is common to all embodiments to which an internal combustion engine controller according to the present invention is applied.","An internal combustion engine 20 comprises a multi-cylinder, in-cylinder fuel injection internal combustion engine.","Its intake system is such that the air coming from the outside passes through an air cleaner 1, an intake manifold 4, and a collector 5 in order named, and flows into each cylinder 9.The amount of air inflow is regulated by an electronic throttle 3.A spark plug 8 and a fuel injection valve 7 are mounted on each cylinder 9.Further, each cylinder 9 is provided with an intake valve 27 and an exhaust valve 28.The employed exhaust system is such that an exhaust manifold 10 is connected to each cylinder 10.A three-way catalyst 11 is mounted on the exhaust manifold 10.Further, an A/F sensor 12 is mounted between each cylinder 9 and the three-way catalyst 11.An exhaust gas re-circulation path (EGR path) 18 is furnished to establish communication between the intake manifold 4 and exhaust manifold 10 by bypassing each cylinder 9.The EGR path is provided with an EGR valve 19.The intake manifold 4 for the intake system is provided with an air flow sensor 2.The air flow sensor detects the amount of air inflow.","A crank angle sensor 15 outputs a signal each time a crankshaft rotates one degree.","A throttle opening sensor 17, which is mounted on an electronic throttle 3, detects the opening of the electronic throttle 3.A water temperature sensor 14 detects a cooling water temperature of the internal combustion engine 20.An accelerator opening sensor 13 detects the depression amount of an accelerator 6 for the purpose of detecting a torque demanded by the driver.","The signals output from the accelerator opening sensor 13, air flow sensor 2, throttle opening sensor 17, crank angle sensor 15, and water temperature sensor 14 are transmitted to a control unit (controller) 50.The information about the operating status of the internal combustion engine 20 is derived from the signals output from these sensors to optimally compute major manipulated variables of the internal combustion engine, such as the intake air volume, fuel injection quantity, and ignition timing.","The fuel injection quantity computed within the control unit 50 is converted to a valve open pulse signal and forwarded to the fuel injection valve 7.Further, the control unit 50 computes an ignition time as specified.","To ensure that ignition takes place at the computed ignition time, the control unit 50 outputs a drive signal to the spark plug 8.The intake air delivered from the intake system is regulated by the electronic throttle 3, mixed with a re-circulated exhaust gas that is regulated by the EGR valve 19, and admitted into the cylinder 9 via the intake valve 27.The fuel injected from the fuel injection valve 7 into the cylinder (combustion chamber) 9 is mixed with an air inflow from the intake manifold 4 to form an air-fuel mixture.","The air-fuel mixture is combustively burned by a spark that is generated from the spark plug 8 at the computed ignition time.","The resulting combustion pressure pushes a piston 29 downward so as to generate motive power for the internal combustion engine 20.The exhaust gas generated upon combustion goes into the three-way catalyst 11 via the exhaust manifold 10.All exhaust gas components (HC, CO, and NOx) are then purified in the three-way catalyst 11 and discharged again to the outside.","The amount of exhaust gas re-circulated to the intake side via an exhaust gas re-circulation pipe 18 is regulated by the EGR valve 19.The A/F sensor 12 has a linear output characteristic in relation to the concentration of oxygen contained in the exhaust gas.","The relationship between the concentration of oxygen in the exhaust gas and the air-fuel ratio is substantially linear.","Therefore, the A/F sensor 12, which detects the concentration of oxygen, can determine the air-fuel ratio of the internal combustion engine 20.The control unit 50 is configured so as to use the signal from the A/F sensor 12 to calculate the upstream air-fuel ratio of the three-way catalyst 11, and exercise feedback control to successively correct the fuel injection quantity or air flow rate for the purpose of ensuring that the air-fuel ratio of an air-fuel mixture in the cylinder 9 for the internal combustion engine 20 agrees with a target air-fuel ratio.","FIG.","2 illustrates the internal configuration of the control unit (ECU) 50 for the internal combustion engine 20 shown in FIG.","1.The values output from the A/F sensor 12, throttle opening sensor 17, air flow sensor 2, internal combustion engine rotating speed sensor 15, and water temperature sensor 14 enter the ECU 50.The output values are then subjected to noise elimination and other signal processes in an input circuit 54, and forwarded to an input/output port 55.The value of the input/output port 55 is stored in a RAM 53 and subjected to arithmetic processing by a CPU 51.A control program containing the description of arithmetic processing is stored in a ROM 52 beforehand.","The values representing the amounts of individual actuator operations, which are computed by the control program, are stored in the RAM 53 and then forwarded to the input/output port 55.An ON/OFF signal is set as the operation signal for the spark plug 8, which is used for ignition and combustion.","This signal is ON when the primary coil in an ignition output circuit 56 is energized and OFF when the same circuit is de-energized.","Ignition takes place when the signal status changes from ON to OFF.","A spark plug signal, which is set for the input/output port 55, is amplified to energy adequate for combustion and then supplied to the spark plug 8.As a drive signal for the fuel injection valve 7, an ON/OFF signal is set.","This signal turns ON to open the valve and turns OFF to close the valve.","The signal is amplified to an adequate energy level that is required for operating the fuel injection valve drive circuit 57 to open the fuel injection valve 7, and then forwarded to the fuel injection valve 7.A drive signal for opening the electronic throttle 3 to a target degree of opening is delivered to the electronic throttle 3 via an electronic throttle drive circuit 58.The configuration common to all embodiments of an internal combustion engine controller according to the present invention has been described.","From now on, individual embodiments will be described.","First Embodiment The control program to be written into the ROM 52 of the EPU 50 will now be described.","FIG.","3 is a control block diagram illustrating the overall control operation that the EPU 50 shown in FIG.","2 performs in accordance with a first embodiment.","The major portion of a fuel-based, torque-on-demand control is shown in FIG.","3.This control comprises a target torque computing section 61, a fuel injection quantity computing section 62, a fuel injection quantity correction section 63, a target equivalence ratio computing section 64, a target air flow rate computing section 65, an actual air flow rate computing section 66, a target throttle opening computing section 67, and a throttle opening control section 68.The target torque computing section 61 computes a target torque TgTc from an accelerator opening Apo and an internal computer combustion engine rotating speed Ne.","The fuel injection quantity computing section 62 operates based on the target torque TgTc to compute a fuel injection quantity TIO that provides the target torque TgTc.","The fuel injection quantity correction section 63 provides phase compensation so that the fuel injection quantity TIO matches the phase of the air in the cylinder 9, and then computes a corrected fuel injection quantity TI.","The target equivalence ratio computing section 64 computes a target equivalence ratio TgFbya from the target torque TgTc and internal combustion engine rotating speed Ne.","For the sake of convenience, the ratio between fuel and air is handled in the form of an equivalence ratio.","It goes without saying that the air-fuel ratio may be used as is.","It should be noted that the target equivalence ratio computing section 64 also chooses between homogeneous combustion and stratified combustion.","The target air flow rate computing section 65 computes a target air flow rate TgTp from the fuel injection quantity TIO and target equivalence ratio TgFbya.","Although this will be described in detail later, the amount of air inflow into a cylinder per cycle is standardized as a value of the target air flow rate TgTp for the sake of convenience.","The actual air flow rate computing section 66 converts a mass flow rate Qa detected by the air flow sensor 2 into an actual air flow rate Tp, which is of the same dimension as the target air flow rate TgTP and flows into a cylinder per cycle, and then outputs the converted value.","The target throttle opening computing section 67 computes a target throttle opening TgTvo in accordance with the target air flow rate TgTp and actual air flow rate Tp.","The throttle opening computing section 68 computes a throttle manipulated variable Tduty from the target throttle opening TgTvo, the actual opening of the electronic throttle 3, and the value Tvo.","The throttle manipulated variable Tduty represents the duty ratio of a PWM signal that enters a drive circuit 58, which controls a throttle motor drive current.","The control computing sections and correction sections of the first embodiment of the control block described above will now be described in detail with reference to FIGS.","4 to 12.1.Target Torque Computing Section FIG.","4 illustrates the target torque computing section 61.The value TgTc represents a torque that is equivalent to a target combustion pressure.","The value TgTa represents a torque required by the accelerator.","The value TgTl represents an air flow rate that is required for idling speed maintenance.","Since an off-idle state provides torque control and an idle state provides output control, it is assumed that the manipulated variable of an idling control means 61a is a theoretical air-fuel ratio, which is proportional to the output.","Gain is provided for use in output-to-torque dimensional conversion.","Further, when the air-conditioner (AC) is ON, the corresponding load is corrected.","If any electrical load exists, the corresponding load is corrected.","For the air flow rate required for idling speed maintenance TgTl, a lower-limit value is defined as a BCV portion for assuring an adequate air flow rate for avoiding a reverse oil flow into the cylinder 6.An idle feedback control means 61a2 functions during an idle state only in order to correct feed-forward-related error.","It is assumed that the controlled variable for the idle feedback control means 61a2 is the rotating speed Ne, and that the manipulated variable is an air flow rate.","The algorithm for the idle feedback control means 61a2 is not specified herein.","However, it may be, for instance, a PID control algorithm.","An idling judgment section 61b concludes that an idle state exists when the accelerator opening Apo is smaller that specified.","It is desirable that the TblTgTa and TblTgTf settings be determined according to actual device data.","2.Fuel Injection Quantity Computing Section FIG.","5 illustrates the fuel injection quantity computing section 62, which converts a target combustion pressure torque TgTc to a fuel injection quantity TIO for control purposes.","The fuel injection quantity TIO is proportional to torque because it represents a fuel injection quantity per cylinder per cycle.","This proportional relationship is used to convert the target combustion pressure torque TgTc to the fuel injection quantity TIO.","A gain may be used, but table conversion may be effected in consideration of some possible error.","It is desirable that the setting be determined according to actual device data.","3.Fuel Injection Quantity Correction Section FIG.","6 illustrates the fuel injection quantity correction section 63, which provides correction to adjust the fuel injection quantity TIO for the phase of the air in the cylinder 9.The air transmission characteristic between the throttle 3 and cylinder 9 is approximated with wasted time plus first-order lag.","It is desirable that the settings for parameter n1, which represents wasted time, and parameter Kair, which corresponds to a time constant for a first-order lag, be determined according to actual device data.","Further, parameter n1 and parameter Kair may be varied depending on the operating conditions.","4.Target Equivalence Ratio Computing Section FIG.","7 illustrates the target equivalence ratio computing section 64, which determines the combustion status and computes the target equivalence ratio.","Fpstratify is a stratified combustion permission flag.","When Fpstratify=1, the injection time, ignition time, injection quantity, and air flow rate are regulated to provide stratified combustion.","Injection time/ignition time determination is not described herein.","The stratified combustion permission flag Fpstratify is 1 to permit stratified combustion if the conditions are satisfied by the water temperature Twn, accelerator opening Apo, and rotating speed Ne.","When stratified combustion is permitted, the value for letting a stratified combustion target equivalence ratio map Mtgfba_s be referenced by the target combustion pressure torque TgTc and rotating speed Ne is the target equivalence ratio TgFbya.","When TgFbya=0, homogeneous combustion takes place, and the value for letting a homogeneous combustion target equivalence ratio map Mtgfba be referenced by the target combustion pressure TgTc and rotating speed Ne is the target equivalence ratio TgFbya.","It is desirable that the settings for the stratified combustion target equivalence ratio map Mtgfba_s and homogeneous combustion target equivalence ratio map Mtgfba be determined according to actual device data.","5.Target Air Flow Rate Computing Section FIG.","8 illustrates the target air flow rate computing section 65, which computes the target air flow rate TgTp.","For the sake of convenience, the target air flow rate TgTp is computed as a standardized value representing the air flow rate that flows into a cylinder per cycle.","As is obvious from FIG.","8, the target air flow rate TgTp is calculated from the fuel injection quantity TIO and target equivalence ratio TgFbya.","The equation used for this calculation is as follows: TgTp=TIO×(1/TgFbya) 6.Actual Air Flow Rate Computing Section FIG.","9 illustrates the actual air flow rate computing section 66, which computes the actual air flow rate Tp.","For the sake of convenience, the actual air flow rate Tp is computed as a standardized value representing the air flow rate that flows into a cylinder per cycle.","The value Qa is an incoming air flow rate that is detected by the air flow sensor 2.The value K is determined so that the actual air flow rate Tp provides the fuel injection quantity prevalent at the theoretical air-fuel ratio.","The value Cy1 denotes the number of cylinders for the internal combustion engine 20.7.Target Throttle Opening Computing Section FIG.","10 illustrates the target throttle opening computing section 67, which determines the target throttle opening TgTVO from the target air flow rate TgTp, actual air flow rate Tp, and rotating speed Ne.","This control block can be divided into a section 67b (second target throttle opening computing means) for exercising feed-forward control to determine the target throttle opening TgTVOFF from the target air flow rate TgTp and rotating speed Ne and a feedback control section 67a (first target throttle opening computing means) for exercising feedback control to determine the target throttle opening TgTVOFB from the target air flow rate TgTp and actual air flow rate Tp.","It is assumed, as shown in FIG.","10, that the feed-forward control section 67b determines the target throttle opening TgTVOFF by referencing a map.","It is desirable that the map setting be determined according to actual device data.","Feedback control is exercised to provide PID control.","Each gain is given as a magnitude of deviation between the target air flow rate TgTp and actual air flow rate TpTg.","However, it is desirable that the setting be determined according to actual device data.","For the D portion, an LPF (low-pass filter) is provided for high-frequency noise elimination.","The target throttle opening TgTVOFF that is computed by the second section 67b of feed-forward control, and the target throttle opening TgTVOFB that is determined by the first section 67a of feedback control are output to the third control section 67c (third throttle opening computing means).","The third control section 67c then computes the sum of the target throttle opening TgTVOFF and target throttle opening TgTVOFB and uses the computed sum as the final target throttle opening TgTVO.","8.Throttle Opening Control Section FIG.","11 illustrates the throttle opening control section 68, which computes the throttle drive manipulated variable Tduty from the target throttle opening TgTVO and actual throttle opening Tvo.","As described earlier, the throttle drive manipulated variable Tduty denotes the duty ratio of a PWM signal that enters a drive circuit 58, which controls a throttle motor drive current.","It is assumed that the throttle drive manipulated variable Tduty is determined by means of PID control.","Although no details will be given, it is desirable that each PID control gain be tuned with an actual device for optimum results.","Second Embodiment The control block of an internal combustion engine controller according to a second embodiment of the present invention will now be described in detail.","FIG.","12 is a control block diagram of the second embodiment that illustrates the overall control of the EPU 50 shown in FIG.","2.The major portion of a fuel-based, torque-on-demand control is shown in FIG.","12.The target torque computing section 61, fuel injection quantity computing section 62, fuel injection quantity correction section 63, target equivalence ratio computing section 64, target air flow rate computing section 65, actual air flow rate computing section 66, and target throttle opening computing section 67 within the control block of the second embodiment are the same as their counterparts described in Sections 1 to 7, respectively, under the first embodiment and will not be described again.","8.Throttle Opening Control Section FIG.","13 illustrates the throttle opening control section 68.As indicated in FIG.","13, feed-forward control is exercised to determine the throttle drive manipulated variable Tduty from the target throttle opening TgTVO only.","The feed-forward control portion may be a gain only.","In such an instance, however, the manipulated variable (Tduty) for throttle opening control is computed directly from a target air flow rate TgTp without computing a target throttle opening.","Third Embodiment The control block of an internal combustion engine controller according to a third embodiment of the present invention will now be described in detail.","The control block of the third embodiment differs from that of the first embodiment, which is shown in FIG.","3, in that the target air flow rate TgTp for use in feedback control is filtered in relation to a feed-forward control target value in the target throttle opening computing section 67.The filter characteristic will be described in detail later.","However, it is a response characteristic between the target throttle opening and in-cylinder actual air flow rate that is computed by means of feed-forward control in the target air flow rate computing section 65.The purpose is to exercise feedback control to compensate for the steady-state deviation.","The target torque computing section 61, fuel injection quantity computing section 62, fuel injection quantity correction section 63, target equivalence ratio computing section 64, target air flow rate computing section 65, actual air flow rate computing section 66, and throttle opening control section within the control block of the third embodiment are the same as their counterparts described in Sections 1 to 6 and 8, respectively, under the first embodiment and will not be described again.","7.Target Throttle Opening Computing Section As shown in FIG.","14, the target air flow rate TgTp for feedback control is a value that is obtained by filtering the target air flow rate TgTp with a filter 67d.","As indicated in FIG.","15, the specifications for the filter 67c are such that the air transmission characteristic between the throttle 3 and cylinder 9 is approximated with wasted time plus first-order lag.","It is desirable that the settings for parameter n2, which represents wasted time, and parameter Kair2, which corresponds to a time constant for a first-order lag, be determined according to actual device data.","Further, the parameter n2 and the parameter Kair2 may be varied depending on the operating conditions.","Fourth Embodiment The control block of an internal combustion engine controller according to a fourth embodiment of the present invention will now be described in detail.","The control block of the fourth embodiment differs from that of the first embodiment, which is shown in FIG.","3, in that the feedback control's computation of target throttle opening TgTVO can be permitted depending on the operating conditions in the target throttle opening computing section 67.The target torque computing section 61, fuel injection quantity computing section 62, fuel injection quantity correction section 63, target equivalence ratio computing section 64, target air flow rate computing section 65, actual air flow rate computing section 66, and throttle opening control section within the control block of the fourth embodiment are the same as their counterparts described in Sections 1 to 6 and 8, respectively, under the first embodiment and will not be described again.","7.Target Throttle Opening Computing Section When the feedback control computation permission flag FPFB is 1 in the feedback control permission judgment section 67e, a selector section 67f effects a changeover, as shown in FIG.","16, so that the target throttle opening TgTVOFB is determined by means of feedback control.","When the feedback control computation permission flag FPFB is 0, on the other hand, the target throttle opening TgTVOFB is set to 0 without exercising feedback control to perform computations.","As indicated in FIG.","17, the feedback control computation permission flag FPFB is established when the accelerator opening Apo and the time elapse after internal combustion engine startup Tafst are as specified and the air flow sensor and throttle are both normal.","The diagnostic check methods for the air flow sensor 2 and throttle 3 are not particularly described herein; however, various practical methods are available.","Fifth Embodiment The control block of an internal combustion engine controller according to a fifth embodiment of the present invention will now be described in detail.","The control block of the fifth embodiment differs from that of the first embodiment, which is shown in FIG.","3, in that the feed-forward control's computation of target throttle opening TgTPVOFF can be permitted depending on the operating conditions in the target throttle opening computing section 67.The target torque computing section 61, fuel injection quantity computing section 62, fuel injection quantity correction section 63, target equivalence ratio computing section 64, target air flow rate computing section 65, actual air flow rate computing section 66, and throttle opening control section within the control block of the fifth embodiment are the same as their counterparts described in Sections 1 to 6 and 8, respectively, under the first embodiment and will not be described again.","7.Target Throttle Opening Computing Section When the feed-forward control computation permission flag FPFB is 1 in the feed-forward control permission judgment section 67g, the target throttle opening TgTVOFF is determined by means of feed-forward control as shown in FIG.","18.When the feed-forward control computation permission flag FPFF is 0, on the other hand, the target throttle opening TgTVOFF is set to 0 without exercising feed-forward control to perform computations.","As indicated in FIG.","19, the feed-forward control computation permission flag FPFF is established when the target EGR rate TgEgr is as specified and the air flow sensor is normal.","The diagnostic check method for the air flow sensor is not particularly described herein.","Sixth Embodiment The control block of an internal combustion engine controller according to a sixth embodiment of the present invention will now be described in detail.","The control block of the sixth embodiment differs from that of the first embodiment, which is shown in FIG.","3, in that feed-forward control is exercised in the target throttle opening computing section 67 to compute the target throttle opening TgTVOFF while considering the exhaust gas re-circulation rate.","The target torque computing section 61, fuel injection quantity computing section 62, fuel injection quantity correction section 63, target equivalence ratio computing section 64, target air flow rate computing section 65, actual air flow rate computing section 66, and throttle opening control section within the control block of the sixth embodiment are the same as their counterparts described in Sections 1 to 6 and 8, respectively, under the first embodiment and will not be described again.","7.Target Throttle Opening Computing Section As indicated in FIG.","20, the target throttle opening TgTVOFF, which is determined by means of feed-forward control, is calculated according to the following equation: TgTVOFF=TgTVOFF0×EgrHos where EgrHOS is a throttle opening correction factor that is used in the event of EGR, and the target throttle opening TgTVOFF0 is determined in the same manner as with the first embodiment.","As indicated in FIG.","20, a map is referenced in accordance with the target combustion pressure torque TgTc and rotating speed Ne to determine the target EGR rate TgEgr.","Further, a map is referenced in accordance with the target EGR rate TgEgr and target air flow rate TgTP to determine the throttle opening correction factor EgrHos.","It is desirable that the settings for the maps be determined according to actual device data.","Seventh Embodiment The control block of an internal combustion engine controller according to a seventh embodiment of the present invention will now be described in detail.","The control block of the seventh embodiment differs from that of the first embodiment, which is shown in FIG.","3, in that feed-forward control is exercised in the target throttle opening computing section 67 to compute the target throttle opening TgTVOFF in accordance with an inverse model for the response characteristic between the air flow rate in the cylinder 9 and throttle 3.Further, the air flow rate in the cylinder 9 is used as the actual air flow rate Tp for feedback control.","The target torque computing section 61, fuel injection quantity computing section 62, fuel injection quantity correction section 63, target equivalence ratio computing section 64, target air flow rate computing section 65, and throttle opening control section within the control block of the seventh embodiment are the same as their counterparts described in Sections 1 to 5 and 8, respectively, under the first embodiment and will not be described again.","6.Actual Air Flow Rate Computing Section As indicated in FIG.","21, the air flow rate Tp1 flowing into a cylinder per cycle is subjected to the air transmission characteristic prevalent between the throttle and cylinder.","The transmission characteristic is approximated with wasted time plus first-order lag.","It is desirable that the settings for parameter n3, which represents wasted time, and parameter Kair3, which corresponds to a time constant for a first-order lag, be determined according to actual device data.","Further, the parameter n3 and the parameter Kair3 may be varied depending on the operating conditions.","7.Target Throttle Opening Computing Section As indicated in FIG.","22, an inverse filter (inverse model) 67k is used to determine the target throttle opening TgTVOFF for feed-forward control.","Detailed specifications for the inverse filter 67k are shown in FIG.","23.It is desirable that the settings for a1, b0, and b1 be determined according to actual device data.","It is also conceivable that the settings may be varied depending on the operating conditions.","Eighth Embodiment The control block of an internal combustion engine controller according to an eighth embodiment of the present invention will now be described in detail.","FIG.","24 is a control block diagram of the eighth embodiment that illustrates the overall control of the EPU 50 shown in FIG.","2.The major portion of a fuel-based, torque-on-demand control is shown in FIG.","24.The control block of the eighth embodiment differs from that of the first embodiment, which is shown in FIG.","3, in that an air-fuel ratio feedback control section 69 is added.","The air-fuel ratio feedback control section 69 provides a manipulated variable Alpha, which is used to correct the target air flow rate TgTP that is computed by the target air flow rate computing section 65.The above functionality is added so as to correct the target air flow rate TgTP in accordance with the value output from the A/F sensor 12, which is mounted on the exhaust manifold 10.The target torque computing section 61, fuel injection quantity computing section 62, fuel injection quantity correction section 63, target equivalence ratio computing section 64, actual air flow rate computing section 66, target throttle opening computing section 67, and throttle opening control section 68 within the control block of the eighth embodiment are the same as their counterparts described in Sections 1 to 4 and 7 to 9, respectively, under the first embodiment and will not be described again.","5.Air-Fuel Ratio Feedback Control Section This section exercises feedback control in accordance with the equivalence ratio Rfbya, which is detected by the A/F sensor 12, so that the internal combustion engine's equivalence ratio agrees with the target equivalence ratio TgFbya under arbitrary operating conditions.","More specifically, PI control is exercised as indicated in FIG.","25 to compute the air-fuel ratio correction term Alpha from the deviation between the target equivalence ratio TgFbya and the equivalence ratio Rfbya detected by the A/F sensor.","6.Target Air Flow Rate Computing Section As shown in FIG.","26, the target air flow rate TgTp is computed from the target equivalence ratio TgFbya and the equivalence ratio Rfbya detected by the A/F sensor.","For computation, the following equation is used: TgTp=TIO×(1/(TgFbya×Alpha)) Although the present invention has been described in detail with reference to the first to eighth embodiments thereof, it will be readily apparent to those skilled in the art that the present invention is not limited to those preferred embodiments, and that various design changes may be made therein without departing from the spirit and scope of the appended claims.","In the target throttle opening computing section, for instance, the target throttle opening computed by the second throttle opening computing means and the target throttle opening computed by the first throttle opening computing means are output to the third throttle opening computing means, which computes the sum of the first and second target throttle opening values and uses the computed sum as the final target throttle opening.","However, the third throttle opening computing means can alternatively compute the final target throttle opening while considering not only the sum but also the product or other control element.","In the foregoing embodiments, an air flow sensor is adopted as means for measuring the air flow rate and used to measure a mass flow rate.","In an alternative embodiment, however, a pressure sensor for measuring the intake pipe internal pressure may also be used.","Industrial Applicability As described above, the internal combustion engine controller according to the present invention comprises a first throttle opening computing means, which computes a throttle opening in accordance with a value detected by an air flow sensor, and a second throttle opening computing means, which computes a throttle opening without using a value detected by an air flow sensor.","Therefore, when used in a fuel-based torque-on-demand control type internal combustion engine, the internal combustion engine controller according to the present invention can improve air flow rate response and convergence, reduce the exhaust gas emission, and provides improved torque response without being affected by various environmental changes or sensor, actuator, and other component variations."]],"string":"[\n [\n \"Controller of internal combustion engine\",\n \"Disclosed is an internal combustion engine controller, which is used with a fuel-based torque-on-demand control type, multi-cylinder, direct-injection internal combustion engine to provide an air flow rate control means that excels in response and convergence.\",\n \"This internal combustion engine controller comprises means for computing a target throttle opening from operating conditions.\",\n \"The means for computing a target throttle opening includes: a first computing means for determining a target throttle opening by exercising feedback control in accordance with operating conditions including an intake air flow rate; a second computing means for determining a target throttle opening by exercising feed-forward control in accordance with operating conditions; and a third computing means for determining a target throttle opening in accordance with a target throttle opening value determined by the first computing means and a target throttle opening value determined by the second computing means.\"\n ],\n [\n \"1.An internal combustion engine controller, comprising: means for computing a target throttle opening from operating conditions; wherein the means for computing a target throttle opening comprises: a first computing means for determining a target throttle opening by exercising feedback control in accordance with operating conditions including an intake air flow rate; a second computing means for determining a target throttle opening by exercising feed-forward control in accordance with operating conditions; and a third computing means for determining a target throttle opening in accordance with a target throttle opening value determined by said first computing means and a target throttle opening value determined by said second computing means.\",\n \"2.An internal combustion engine controller, comprising: a target air flow rate computing means for computing a target air flow rate in accordance with operating conditions; a first computing means for determining a target throttle opening by exercising feedback control in accordance with a difference between said target air flow rate and an actual air flow rate; a second computing means for determining a target throttle opening by exercising feed-forward control in accordance with said target air flow rate; and a third computing means for determining a target throttle opening in accordance with a target throttle opening value determined by said first computing means and a target throttle opening value determined by said second computing means.\",\n \"3.An internal combustion engine controller, comprising: a control unit for computing a target throttle opening from operating conditions; wherein the control unit comprises: a first computing means for determining a target throttle opening by exercising feedback control in accordance with operating conditions including an intake air flow rate; a second computing means for determining a target throttle opening by exercising feed-forward control in accordance with operating conditions; a third computing means for determining a target throttle opening in accordance with a target throttle opening value determined by said first computing means and a target throttle opening value determined by said second computing means; and a throttle opening control means for driving a throttle in accordance with a target throttle opening calculated by said third computing means.\",\n \"4.The internal combustion engine controller according to claim 3, wherein said throttle opening control means comprises means for controlling a throttle in such a manner as to reduce a difference between a target throttle opening calculated by said third computing means and a directly or indirectly detected actual throttle opening.\",\n \"5.The internal combustion engine controller according to claim 1, wherein said target throttle opening computing means comprises a computation permission means for permitting said first target throttle opening computing means to compute a target throttle opening.\",\n \"6.The internal combustion engine controller according to claim 5, wherein said computation permission means inhibits said first target throttle opening computing means from computing a target throttle opening if means for detecting an actual air flow rate directly or indirectly is found to be abnormal; and wherein said third target throttle opening computing means calculates a target throttle opening in accordance with a second target throttle opening and makes it possible to adjust the actual air flow rate for a target air flow rate.\",\n \"7.The internal combustion engine controller according to claim 5, wherein said computation permission means inhibits said first target throttle opening computing means from computing a target throttle opening if an accelerator opening is not smaller than a predetermined value or said second target throttle opening is not smaller than a predetermined value; and wherein said third target throttle opening computing means calculates a target throttle opening in accordance with the second target throttle opening and makes it possible to adjust an actual air flow rate to a target air flow rate.\",\n \"8.The internal combustion engine controller according to claim 5, wherein said computation permission means inhibits said first target throttle opening computing means from calculating a target throttle opening if a difference between a target air flow rate and an actual air flow rate is not greater than a predetermined value; and wherein said third target throttle opening computing means calculates a target throttle opening in accordance with the second target throttle opening and makes it possible to adjust an actual air flow rate to a target air flow rate.\",\n \"9.The internal combustion engine controller according to claim 1, wherein said target throttle opening computing means comprises a computation permission means for permitting said second target throttle opening computing means to compute a target throttle opening in accordance with operating conditions.\",\n \"10.The internal combustion engine controller according to claim 9, wherein said computation permission means inhibits said second target throttle opening computing means from computing a target throttle opening if a target exhaust gas re-circulation rate is not lower than a predetermined value; and wherein said third target throttle opening computing means calculates a target throttle opening in accordance with a first target opening and makes it possible to adjust an actual air flow rate to a target air flow rate.\",\n \"11.The internal combustion engine controller according to claim 1, wherein said target throttle opening computing means makes it possible to adjust an actual air flow rate to a target air flow rate by calculating said third target throttle opening from said first target throttle opening, which is computed in accordance with an actual air flow rate, even if a relationship between said second target throttle opening and the actual air flow rate is significantly changed.\",\n \"12.The internal combustion engine controller according to claim 2, wherein said target throttle opening computing means comprises a target air flow rate correction means for correcting said target air flow rate; and wherein said first target throttle opening computing means computes a first target throttle opening in accordance with a difference between an actual air flow rate and a target air flow rate calculated by said target air flow rate correction means.\",\n \"13.The internal combustion engine controller according to claim 12, wherein said target air flow rate correction means represents a response characteristic between said second target throttle opening and an actual air flow rate.\",\n \"14.The internal combustion engine controller according to claim 1, wherein said second target throttle opening computing means computes a second target throttle opening in accordance with a rotating speed of an internal combustion engine and a target air flow rate.\",\n \"15.The internal combustion engine controller according to claim 1, further comprising means for computing a target exhaust gas re-circulation rate; wherein said second target throttle opening computing means computes a second target throttle opening in accordance with a rotating speed of an internal combustion engine, a target air flow rate, and the target exhaust gas re-circulation rate.\",\n \"16.The internal combustion engine controller according to claim 2, wherein said target air flow rate computing means computes said target air flow rate in accordance with a target air-fuel ratio and a target fuel injection quantity.\",\n \"17.The internal combustion engine controller according to claim 2, further comprising: detection means for detecting exhaust gas components of an internal combustion engine directly or indirectly; and a correction amount computing means for computing an amount of correction for said target air flow rate in accordance with the exhaust gas components detected by said detection means.\",\n \"18.The internal combustion engine controller according to claim 1, wherein said second target throttle opening computing means computes a second target throttle opening in accordance with an inverse model for a response between an in-cylinder target air flow rate and a throttle opening.\",\n \"19.The internal combustion engine controller according to claim 1, wherein the actual air flow rate is a value obtained by detecting an in-cylinder air flow rate directly or indirectly.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>In recent years, worldwide efforts have been made to provide increased energy savings.\",\n \"In the field of automotive technology, the development of a fuel-efficient internal combustion engine has been required for energy saving purposes.\",\n \"The most conspicuous internal combustion engine meeting such a demand is a lean-burn internal combustion engine.\",\n \"In particular, an in-cylinder fuel injection internal combustion engine, which is a lean-burn internal combustion engine, injects fuel directly into a cylinder to stratify an air-fuel mixture, thereby making it possible to achieve combustion at an air-fuel ratio of higher than 40 and reduce the pump loss.\",\n \"In a lean-burn in-cylinder fuel injection internal combustion engine system, which is described above, the air flow rate is not proportional to the torque.\",\n \"Therefore, the lean-burn in-cylinder fuel injection internal combustion engine system generally uses an electronic throttle for electronically controlling the air flow rate unlike a conventional internal combustion engine system.\",\n \"For the above lean-burn in-cylinder fuel injection internal combustion engine system, torque-on-demand control is required to provide torque desired by the driver at a wide-range air-fuel ratio.\",\n \"Two types of torque-on-demand control are an air-based type and a fuel-based type.\",\n \"If air-based torque-on-demand control is exercised, a target torque computation section and a target air-fuel ratio computation section determine a target torque and target air-fuel ratio, respectively, as shown in FIG.\",\n \"27 .\",\n \"A target air flow rate computation section for providing the target torque and target air-fuel ratio computes a target air flow rate.\",\n \"An electronic throttle controls the air flow rate.\",\n \"An air flow rate sensor detects an actual air flow rate.\",\n \"A fuel injection quantity computation section determines the quantity of fuel injection from the actual air flow rate and target air-fuel ratio.\",\n \"If, on the other hand, fuel-based torque-on-demand control is exercised, a target torque computation section determines a target torque as shown in FIG.\",\n \"28 .\",\n \"A fuel injection quantity computation section then determines the quantity of fuel injection for providing the target torque.\",\n \"A target air computation section computes a target air flow rate from the fuel injection quantity and target air-fuel ratio.\",\n \"An electronic throttle controls the air flow rate.\",\n \"Further, fuel-based torque-on-demand control can be exercised to provide feedback control over the air flow rate in accordance with a value output by an air flow sensor.\",\n \"Fuel-based torque-on-demand control described above uses an electronic throttle to exercise air flow rate control after fuel injection quantity determination.\",\n \"However, a transmission characteristic exists between the electronic throttle and cylinder.\",\n \"More specifically, a transient phenomenon occurs because it generally takes tens to hundreds of milliseconds for the air flow rate controlled near the electronic throttle to arrive in a cylinder as shown in FIG.\",\n \"29 .\",\n \"In an in-cylinder fuel injection internal combustion engine, on the other hand, fuel injection directly occurs within a cylinder.\",\n \"Therefore, the transmission characteristic of the fuel injection side is smaller than that of the air side because time is merely wasted by intermittent combustion.\",\n \"Meanwhile, the exhaust pipe for an internal combustion engine is usually provided with a three-way catalyst or a catalyst having a three-way catalytic function as an exhaust gas emission purification system.\",\n \"As shown in FIG.\",\n \"30 , the three-way catalyst efficiently purifies carbon hydride (HC), carbon monoxide (CO), which are reducers, and nitrogen oxide (NOx), which is an oxidant, only in the neighborhood of a theoretical air-fuel ratio.\",\n \"From the viewpoint of exhaust gas emission reduction, it is desirable that the air-fuel ratio for an internal combustion engine be adjusted for the theoretical air-fuel ratio.\",\n \"As regards a lean air-fuel ratio, there is a correlation between the air fuel ratio and internal combustion engine combustion stability as shown in FIG.\",\n \"31 .\",\n \"It is therefore necessary to control the air fuel ratio for the purpose of providing combustion stability of an internal combustion engine.\",\n \"Thus, when an air flow rate transient phenomenon in a cylinder is considered, the fuel injection quantity needs to be controlled from the viewpoint of exhaust gas emission reduction for theoretical air-fuel ratio or from the viewpoint of internal combustion engine combustion stability for lean air-fuel ratio.\",\n \"Further, the torque of an internal combustion engine is determined almost conclusively by the fuel injection quantity.\",\n \"Therefore, the torque-response is determined by the transient characteristic of air flow rate.\",\n \"As is obvious from the above description, the most important tasks to be accomplished for an internal combustion engine that provides fuel-based torque-on-demand control are to improve the response to air flow rate control and convergence performance, minimize the variations among mass-produced products, and improve the robustness for changes with time.\",\n \"A technology disclosed by JP-A No.\",\n \"2000-97086 provides control over the air-fuel ratio of an internal combustion engine.\",\n \"When the target air flow rate changes, this technology provides delay compensation to improve the air flow rate response within a cylinder by temporarily permitting the throttle opening to overshoot the degree of throttle opening for achieving the target air flow rate.\",\n \"However, this control method cannot exercise air flow rate control if the air flow sensor is faulty because it computes the target throttle opening in accordance with the deviation between the actual air flow rate and target air flow rate.\",\n \"Another technology disclosed by JP-A No.\",\n \"6-146950 changes the throttle opening by a predetermined amount, if there is any change in the target air-fuel ratio, to eliminate any inadequate feedback response portion of the actual air flow rate with a view toward response improvement.\",\n \"However, since this control method does not detect the actual air flow rate, it cannot properly respond, for instance, to air density changes at a high altitude and exhibits low robustness for air flow rate control accuracy in relation to various environmental changes such as throttle control sensor and actuator characteristic variations.\",\n \"In consideration of the problems described above, it is an object of the present invention to provide an internal combustion engine controller that is capable of exercising high-performance air flow rate control of a fuel-based torque-on-demand control type, in-cylinder fuel injection internal combustion engine while providing improved response and convergence and enhanced robustness.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>To achieve the above object, an internal combustion engine controller of the present invention basically comprises means for computing a target throttle opening in accordance with operating conditions.\",\n \"The means for computing a target throttle opening comprises a first computing means, which determines a target throttle opening by exercising feedback control in accordance with operating conditions including an intake air volume; a second computing means, which determines a target throttle opening by exercising feed-forward control in accordance with the operating conditions; and a third computing means, which determines a target throttle opening in accordance with the respective target throttle opening values determined by the first computing means and second computing means.\",\n \"In another aspect of the present invention, an internal combustion engine controller comprises: a target air flow rate computing means for computing a target air flow rate in accordance with operating conditions; a first computing means for determining a target throttle opening by exercising feedback control in accordance with a difference between the above target air flow rate and an actual air flow rate; a second computing means for determining a target throttle opening by exercising feed-forward control in accordance with the above target air flow rate; and a third computing means for determining a target throttle opening in accordance with the respective target throttle opening values determined by the first computing means and second computing means.\",\n \"In still another aspect of the present invention, an internal combustion engine controller for computing a target throttle opening in accordance with operating conditions comprises: a first computing means for determining a target throttle opening by exercising feedback control in accordance with operating conditions including the intake air volume; a second computing means for determining a target throttle opening by exercising feed-forward control in accordance with the operating conditions; a third computing means for determining a target throttle opening in accordance with the target throttle opening values determined by the first computing means and second computing means; and a throttle opening control means for driving a throttle valve in accordance with the target throttle calculated by the third computing means.\",\n \"As indicated by the configuration described above, the internal combustion engine controller of the present invention comprises a first throttle opening computing means for computing a throttle opening by exercising feedback control (F/B control) in accordance with the actual air flow rate, target air flow rate, and other operating conditions and a second throttle opening computing means for computing a throttle opening by exercising feed-forward control (F/F control) in accordance with the target air flow rate and other operating conditions.\",\n \"Therefore, the internal combustion engine controller of the present invention can improve the air flow rate calculation response and convergence, reduce the exhaust gas emission, and provides improved torque response without being affected by various environmental changes or sensor, actuator, and other component variations.\",\n \"In other words, the internal combustion engine controller of the present invention is particularly suitable for a fuel-based torque-on-demand control type, internal combustion engine.\",\n \"It is basically characterized by comprising a second throttle opening computing means for computing a target throttle opening from a target air flow rate and a first throttle opening computing means for computing a target throttle opening from the deviation between a target air flow rate and actual air flow rate.\",\n \"Common feedback control computes a manipulated variable in accordance with the resulting controlled variable.\",\n \"It is therefore said that the response of feedback control is theoretically inferior to that of feed-forward control.\",\n \"On the other hand, it is said that feed-forward control excels in response but exhibits low robustness against disturbance.\",\n \"The disadvantages of these two control functions are offset by the internal combustion engine controller of the present invention.\",\n \"Concisely, the internal combustion engine controller of the present invention provides a target throttle opening computing section that is capable of exercising feed-forward control, which is highly responsive, and feedback control, which is robust against disturbance.\",\n \"Consequently, the internal combustion engine controller of the present invention provides not only improved response but also enhanced robustness against disturbance particularly in air flow rate control over a fuel-based torque-on-demand control type, internal combustion engine.\",\n \"To achieve another object, the internal combustion engine controller of the present invention uses the throttle opening control means, which comprises means for controlling the throttle valve in such a manner as to reduce the difference between a directly or indirectly detected actual throttle opening value and a target throttle opening value calculated by the third computing means.\",\n \"As indicated by the configuration described above, the internal combustion engine controller of the present invention provides feedback control, which computes the manipulated variable of the throttle in accordance with the difference between the actual throttle opening and a target throttle opening calculated by the third target throttle opening computing means.\",\n \"Therefore, the manipulated variable for throttle drive can be computed from the target throttle opening and actual throttle opening.\",\n \"It means that the manipulated variable for throttle drive indicates a duty ratio that is entered into a drive circuit for controlling a throttle motor drive current.\",\n \"Thus, the manipulated variable for throttle drive can be accurately calculated by means of PID control.\",\n \"In addition to feedback control, the internal combustion engine controller of the present invention also provides feed-forward control, which computes the manipulated variable of the throttle in accordance with only the target throttle opening calculated by the third target throttle opening computing means.\",\n \"Therefore, computation control can be exercised with high responsiveness.\",\n \"The feed-forward control portion may be a gain only.\",\n \"In such an instance, the manipulated variable for throttle opening control can be computed directly from a target air flow rate without computing a target throttle opening.\",\n \"To achieve another object, the internal combustion engine controller of the present invention uses the target throttle opening computing means, which comprises a computation permission means for permitting the first target throttle opening computing means to compute a target throttle opening.\",\n \"The computation permission means inhibits the first target throttle opening computing means from computing a target throttle opening if the means for detecting the actual air flow rate directly or indirectly is found to be abnormal.\",\n \"The third target throttle opening computing means calculates a target throttle opening in accordance with the second target throttle opening so that the actual air flow rate can be adjusted for the target air flow rate.\",\n \"As indicated in the configuration described above, the internal combustion engine controller of the present invention comprises a computation permission means for permitting the first target throttle opening computing means to compute a target throttle opening.\",\n \"Therefore, the first target throttle opening computing means can be permitted to compute a target throttle opening when, for instance, the opening of an internal combustion engine accelerator is normal, the air flow sensor function is normal, the throttle is normal, or the elapsed time from startup is as specified.\",\n \"It means that target throttle opening computations can be performed in response to changes in the internal combustion engine environment.\",\n \"Even when the means for detecting the air flow rate directly or indirectly is found to be abnormal, it is possible to inhibit the first target throttle opening computing means from computing the first target throttle opening, cause the second target throttle opening computing means to calculate the second target throttle opening as the third target throttle opening, and control the actual air flow rate in accordance with the third target throttle opening.\",\n \"That is, if the means for detecting the air flow rate of the air flow sensor or the like is faulty, the actual air flow rate can be controlled by inhibiting the target throttle opening from being computed by feedback control and using the target throttle opening computed by feed-forward control as the target opening.\",\n \"Therefore, the air flow rate can be preferably controlled even when the air flow sensor or the like is faulty.\",\n \"To achieve another object, the internal combustion engine controller of the present invention uses the computation permission means to inhibit the first target throttle opening computing means from computing the target throttle opening if the accelerator opening is not smaller than a predetermined value or the second target throttle opening is not smaller than a predetermined value.\",\n \"The third target throttle opening computing means computes the target throttle opening in accordance with the second target throttle opening and makes it possible to adjust the actual air flow rate for the target air flow rate.\",\n \"The computation permission means inhibits the first target throttle opening computing means from computing the target throttle opening if the difference between the target air flow rate and actual air flow rate is not greater than a predetermined value.\",\n \"The third target throttle opening computing means calculates the target throttle opening in accordance with the second target throttle opening and makes it possible to adjust the actual air flow rate to the target air flow rate.\",\n \"To achieve another object, the internal combustion engine controller of the present invention uses the target throttle opening computing means, which comprises a computation permission means for permitting the second target throttle opening computing means to compute the target throttle opening in accordance with the operating conditions.\",\n \"The computation permission means inhibits the second target throttle opening computing means from computing the target throttle opening if a target exhaust gas re-circulation rate is not smaller than a predetermined value.\",\n \"The third target throttle opening computing means calculates the target throttle opening in accordance with the first target opening and makes it possible to adjust the actual air flow rate to the target air flow rate.\",\n \"As indicated in the configuration described above, the internal combustion engine controller of the present invention comprises a second computation permission means, which permits the second target throttle opening computing means to compute a throttle opening.\",\n \"Therefore, if, for instance, the EGR rate is as specified or the air flow sensor is normal, the second target throttle opening computing means can be permitted to computer the target throttle opening.\",\n \"Thus, it is possible to compute the target throttle opening that is responsive to changes in the internal combustion engine environment.\",\n \"To achieve another object, the internal combustion engine controller of the present invention uses the target throttle opening computing means to calculate the third target throttle opening from the first target throttle opening, which is computed in accordance with the actual air flow rate even when the relationship between the second target throttle opening and actual air flow rate significantly changes, and makes it possible to adjust the actual air flow rate for the target air flow rate.\",\n \"As indicated in the configuration described above, the internal combustion engine controller of the present invention exercises feedback control to provide computation control so as to adjust the actual air flow rate for the target air flow rate and calculate the target throttle opening even when the relationship between the target throttle opening computed by feed-forward control and the actual air flow rate significantly changes due to an air density change, foreign matter inclusion in an intake pipe, or other environmental change.\",\n \"Therefore, the air flow rate can be accurately controlled in response to the above environmental changes.\",\n \"To achieve another object, the internal combustion engine controller of the present invention uses the target throttle opening computing means, which comprises a target air flow rate correction means for correcting the target air flow rate.\",\n \"The first target throttle opening computing means computes the first target throttle opening in accordance with the difference between the target air flow rate calculated by the target air flow rate correction means and the actual air flow rate.\",\n \"The target air flow rate correction means represents a response characteristic between the second target throttle opening and actual air flow rate.\",\n \"As indicated by the configuration described above, the internal combustion engine controller of the present invention comprises the target air flow rate correction means.\",\n \"Therefore, it can accurately compute a target air flow rate while improving the transmission characteristic by compensating for a time delay involved in intake air flow from the throttle to a cylinder.\",\n \"To achieve another object, the internal combustion engine controller of the present invention uses the second target throttle opening computing means to compute the second target throttle opening in accordance with the internal combustion engine's rotating speed and the target air flow rate.\",\n \"As indicated by the configuration described above, the internal combustion engine controller of the present invention can exercise feed-forward control to compute the second target throttle opening, for instance, from a map depicting the relationship between the internal combustion engine's rotating speed and the target air flow rate.\",\n \"To achieve another object, the internal combustion engine controller of the present invention comprises means for computing the target exhaust gas re-circulation rate.\",\n \"Further, the second target throttle opening computing means computes the second target throttle opening in accordance with the internal combustion engine's rotating speed, the target air flow rate, and the target exhaust gas re-circulation rate.\",\n \"To achieve another object, the internal combustion engine controller of the present invention uses the target air flow rate computing means to compute the target air flow rate in accordance with the target air-fuel ratio and target fuel injection quantity.\",\n \"As indicated by the configuration described above, the internal combustion engine controller of the present invention can correct the second target throttle opening in accordance with the exhaust gas re-circulation rate when the EGR gas recirculates.\",\n \"More specifically, it can determine the target EGR rate from, for instance, a map depicting the relationship between a target combustion pressure torque and internal combustion engine's rotating speed, determine a throttle opening correction factor from, for instance, a map depicting the relationship between the target EGR rate and target air flow rate, and correct the second target throttle opening.\",\n \"To achieve another object, the internal combustion engine controller of the present invention comprises a detection means for detecting the internal combustion engine's exhaust gas components directly or indirectly, and a correction amount computing means for computing the correction amount for the target air flow rate in accordance with the exhaust gas components detected by the detection means.\",\n \"As indicated by the configuration described above, the internal combustion engine controller of the present invention corrects the target air flow rate with a correction amount (manipulated variable) that is used to compute (by means of air-fuel ratio feedback control) the correction amount for the target air flow rate in accordance with a signal from an A/F sensor or other exhaust gas component detection means.\",\n \"It is therefore possible to calculate a target air flow rate with increased accuracy and compute a target throttle opening.\",\n \"To achieve another object, the internal combustion engine controller of the present invention uses the second target throttle opening computing means to compute the second target throttle opening in accordance with an inverse model for the response between an in-cylinder target air flow rate and the throttle.\",\n \"To achieve still another object, the internal combustion engine controller of the present invention uses an in-cylinder air flow rate value, which is detected directly or indirectly, as an actual air flow rate value.\"\n ],\n [\n \"FIELD OF THE INVENTION The present invention related to internal combustion engine controllers, and more particularly to an internal combustion engine controller for accurately adjusting the air flow rate by controlling the air flow rate with an electronic control valve.\",\n \"BACKGROUND OF THE INVENTION In recent years, worldwide efforts have been made to provide increased energy savings.\",\n \"In the field of automotive technology, the development of a fuel-efficient internal combustion engine has been required for energy saving purposes.\",\n \"The most conspicuous internal combustion engine meeting such a demand is a lean-burn internal combustion engine.\",\n \"In particular, an in-cylinder fuel injection internal combustion engine, which is a lean-burn internal combustion engine, injects fuel directly into a cylinder to stratify an air-fuel mixture, thereby making it possible to achieve combustion at an air-fuel ratio of higher than 40 and reduce the pump loss.\",\n \"In a lean-burn in-cylinder fuel injection internal combustion engine system, which is described above, the air flow rate is not proportional to the torque.\",\n \"Therefore, the lean-burn in-cylinder fuel injection internal combustion engine system generally uses an electronic throttle for electronically controlling the air flow rate unlike a conventional internal combustion engine system.\",\n \"For the above lean-burn in-cylinder fuel injection internal combustion engine system, torque-on-demand control is required to provide torque desired by the driver at a wide-range air-fuel ratio.\",\n \"Two types of torque-on-demand control are an air-based type and a fuel-based type.\",\n \"If air-based torque-on-demand control is exercised, a target torque computation section and a target air-fuel ratio computation section determine a target torque and target air-fuel ratio, respectively, as shown in FIG.\",\n \"27.A target air flow rate computation section for providing the target torque and target air-fuel ratio computes a target air flow rate.\",\n \"An electronic throttle controls the air flow rate.\",\n \"An air flow rate sensor detects an actual air flow rate.\",\n \"A fuel injection quantity computation section determines the quantity of fuel injection from the actual air flow rate and target air-fuel ratio.\",\n \"If, on the other hand, fuel-based torque-on-demand control is exercised, a target torque computation section determines a target torque as shown in FIG.\",\n \"28.A fuel injection quantity computation section then determines the quantity of fuel injection for providing the target torque.\",\n \"A target air computation section computes a target air flow rate from the fuel injection quantity and target air-fuel ratio.\",\n \"An electronic throttle controls the air flow rate.\",\n \"Further, fuel-based torque-on-demand control can be exercised to provide feedback control over the air flow rate in accordance with a value output by an air flow sensor.\",\n \"Fuel-based torque-on-demand control described above uses an electronic throttle to exercise air flow rate control after fuel injection quantity determination.\",\n \"However, a transmission characteristic exists between the electronic throttle and cylinder.\",\n \"More specifically, a transient phenomenon occurs because it generally takes tens to hundreds of milliseconds for the air flow rate controlled near the electronic throttle to arrive in a cylinder as shown in FIG.\",\n \"29.In an in-cylinder fuel injection internal combustion engine, on the other hand, fuel injection directly occurs within a cylinder.\",\n \"Therefore, the transmission characteristic of the fuel injection side is smaller than that of the air side because time is merely wasted by intermittent combustion.\",\n \"Meanwhile, the exhaust pipe for an internal combustion engine is usually provided with a three-way catalyst or a catalyst having a three-way catalytic function as an exhaust gas emission purification system.\",\n \"As shown in FIG.\",\n \"30, the three-way catalyst efficiently purifies carbon hydride (HC), carbon monoxide (CO), which are reducers, and nitrogen oxide (NOx), which is an oxidant, only in the neighborhood of a theoretical air-fuel ratio.\",\n \"From the viewpoint of exhaust gas emission reduction, it is desirable that the air-fuel ratio for an internal combustion engine be adjusted for the theoretical air-fuel ratio.\",\n \"As regards a lean air-fuel ratio, there is a correlation between the air fuel ratio and internal combustion engine combustion stability as shown in FIG.\",\n \"31.It is therefore necessary to control the air fuel ratio for the purpose of providing combustion stability of an internal combustion engine.\",\n \"Thus, when an air flow rate transient phenomenon in a cylinder is considered, the fuel injection quantity needs to be controlled from the viewpoint of exhaust gas emission reduction for theoretical air-fuel ratio or from the viewpoint of internal combustion engine combustion stability for lean air-fuel ratio.\",\n \"Further, the torque of an internal combustion engine is determined almost conclusively by the fuel injection quantity.\",\n \"Therefore, the torque-response is determined by the transient characteristic of air flow rate.\",\n \"As is obvious from the above description, the most important tasks to be accomplished for an internal combustion engine that provides fuel-based torque-on-demand control are to improve the response to air flow rate control and convergence performance, minimize the variations among mass-produced products, and improve the robustness for changes with time.\",\n \"A technology disclosed by JP-A No.\",\n \"2000-97086 provides control over the air-fuel ratio of an internal combustion engine.\",\n \"When the target air flow rate changes, this technology provides delay compensation to improve the air flow rate response within a cylinder by temporarily permitting the throttle opening to overshoot the degree of throttle opening for achieving the target air flow rate.\",\n \"However, this control method cannot exercise air flow rate control if the air flow sensor is faulty because it computes the target throttle opening in accordance with the deviation between the actual air flow rate and target air flow rate.\",\n \"Another technology disclosed by JP-A No.\",\n \"6-146950 changes the throttle opening by a predetermined amount, if there is any change in the target air-fuel ratio, to eliminate any inadequate feedback response portion of the actual air flow rate with a view toward response improvement.\",\n \"However, since this control method does not detect the actual air flow rate, it cannot properly respond, for instance, to air density changes at a high altitude and exhibits low robustness for air flow rate control accuracy in relation to various environmental changes such as throttle control sensor and actuator characteristic variations.\",\n \"In consideration of the problems described above, it is an object of the present invention to provide an internal combustion engine controller that is capable of exercising high-performance air flow rate control of a fuel-based torque-on-demand control type, in-cylinder fuel injection internal combustion engine while providing improved response and convergence and enhanced robustness.\",\n \"SUMMARY OF THE INVENTION To achieve the above object, an internal combustion engine controller of the present invention basically comprises means for computing a target throttle opening in accordance with operating conditions.\",\n \"The means for computing a target throttle opening comprises a first computing means, which determines a target throttle opening by exercising feedback control in accordance with operating conditions including an intake air volume; a second computing means, which determines a target throttle opening by exercising feed-forward control in accordance with the operating conditions; and a third computing means, which determines a target throttle opening in accordance with the respective target throttle opening values determined by the first computing means and second computing means.\",\n \"In another aspect of the present invention, an internal combustion engine controller comprises: a target air flow rate computing means for computing a target air flow rate in accordance with operating conditions; a first computing means for determining a target throttle opening by exercising feedback control in accordance with a difference between the above target air flow rate and an actual air flow rate; a second computing means for determining a target throttle opening by exercising feed-forward control in accordance with the above target air flow rate; and a third computing means for determining a target throttle opening in accordance with the respective target throttle opening values determined by the first computing means and second computing means.\",\n \"In still another aspect of the present invention, an internal combustion engine controller for computing a target throttle opening in accordance with operating conditions comprises: a first computing means for determining a target throttle opening by exercising feedback control in accordance with operating conditions including the intake air volume; a second computing means for determining a target throttle opening by exercising feed-forward control in accordance with the operating conditions; a third computing means for determining a target throttle opening in accordance with the target throttle opening values determined by the first computing means and second computing means; and a throttle opening control means for driving a throttle valve in accordance with the target throttle calculated by the third computing means.\",\n \"As indicated by the configuration described above, the internal combustion engine controller of the present invention comprises a first throttle opening computing means for computing a throttle opening by exercising feedback control (F/B control) in accordance with the actual air flow rate, target air flow rate, and other operating conditions and a second throttle opening computing means for computing a throttle opening by exercising feed-forward control (F/F control) in accordance with the target air flow rate and other operating conditions.\",\n \"Therefore, the internal combustion engine controller of the present invention can improve the air flow rate calculation response and convergence, reduce the exhaust gas emission, and provides improved torque response without being affected by various environmental changes or sensor, actuator, and other component variations.\",\n \"In other words, the internal combustion engine controller of the present invention is particularly suitable for a fuel-based torque-on-demand control type, internal combustion engine.\",\n \"It is basically characterized by comprising a second throttle opening computing means for computing a target throttle opening from a target air flow rate and a first throttle opening computing means for computing a target throttle opening from the deviation between a target air flow rate and actual air flow rate.\",\n \"Common feedback control computes a manipulated variable in accordance with the resulting controlled variable.\",\n \"It is therefore said that the response of feedback control is theoretically inferior to that of feed-forward control.\",\n \"On the other hand, it is said that feed-forward control excels in response but exhibits low robustness against disturbance.\",\n \"The disadvantages of these two control functions are offset by the internal combustion engine controller of the present invention.\",\n \"Concisely, the internal combustion engine controller of the present invention provides a target throttle opening computing section that is capable of exercising feed-forward control, which is highly responsive, and feedback control, which is robust against disturbance.\",\n \"Consequently, the internal combustion engine controller of the present invention provides not only improved response but also enhanced robustness against disturbance particularly in air flow rate control over a fuel-based torque-on-demand control type, internal combustion engine.\",\n \"To achieve another object, the internal combustion engine controller of the present invention uses the throttle opening control means, which comprises means for controlling the throttle valve in such a manner as to reduce the difference between a directly or indirectly detected actual throttle opening value and a target throttle opening value calculated by the third computing means.\",\n \"As indicated by the configuration described above, the internal combustion engine controller of the present invention provides feedback control, which computes the manipulated variable of the throttle in accordance with the difference between the actual throttle opening and a target throttle opening calculated by the third target throttle opening computing means.\",\n \"Therefore, the manipulated variable for throttle drive can be computed from the target throttle opening and actual throttle opening.\",\n \"It means that the manipulated variable for throttle drive indicates a duty ratio that is entered into a drive circuit for controlling a throttle motor drive current.\",\n \"Thus, the manipulated variable for throttle drive can be accurately calculated by means of PID control.\",\n \"In addition to feedback control, the internal combustion engine controller of the present invention also provides feed-forward control, which computes the manipulated variable of the throttle in accordance with only the target throttle opening calculated by the third target throttle opening computing means.\",\n \"Therefore, computation control can be exercised with high responsiveness.\",\n \"The feed-forward control portion may be a gain only.\",\n \"In such an instance, the manipulated variable for throttle opening control can be computed directly from a target air flow rate without computing a target throttle opening.\",\n \"To achieve another object, the internal combustion engine controller of the present invention uses the target throttle opening computing means, which comprises a computation permission means for permitting the first target throttle opening computing means to compute a target throttle opening.\",\n \"The computation permission means inhibits the first target throttle opening computing means from computing a target throttle opening if the means for detecting the actual air flow rate directly or indirectly is found to be abnormal.\",\n \"The third target throttle opening computing means calculates a target throttle opening in accordance with the second target throttle opening so that the actual air flow rate can be adjusted for the target air flow rate.\",\n \"As indicated in the configuration described above, the internal combustion engine controller of the present invention comprises a computation permission means for permitting the first target throttle opening computing means to compute a target throttle opening.\",\n \"Therefore, the first target throttle opening computing means can be permitted to compute a target throttle opening when, for instance, the opening of an internal combustion engine accelerator is normal, the air flow sensor function is normal, the throttle is normal, or the elapsed time from startup is as specified.\",\n \"It means that target throttle opening computations can be performed in response to changes in the internal combustion engine environment.\",\n \"Even when the means for detecting the air flow rate directly or indirectly is found to be abnormal, it is possible to inhibit the first target throttle opening computing means from computing the first target throttle opening, cause the second target throttle opening computing means to calculate the second target throttle opening as the third target throttle opening, and control the actual air flow rate in accordance with the third target throttle opening.\",\n \"That is, if the means for detecting the air flow rate of the air flow sensor or the like is faulty, the actual air flow rate can be controlled by inhibiting the target throttle opening from being computed by feedback control and using the target throttle opening computed by feed-forward control as the target opening.\",\n \"Therefore, the air flow rate can be preferably controlled even when the air flow sensor or the like is faulty.\",\n \"To achieve another object, the internal combustion engine controller of the present invention uses the computation permission means to inhibit the first target throttle opening computing means from computing the target throttle opening if the accelerator opening is not smaller than a predetermined value or the second target throttle opening is not smaller than a predetermined value.\",\n \"The third target throttle opening computing means computes the target throttle opening in accordance with the second target throttle opening and makes it possible to adjust the actual air flow rate for the target air flow rate.\",\n \"The computation permission means inhibits the first target throttle opening computing means from computing the target throttle opening if the difference between the target air flow rate and actual air flow rate is not greater than a predetermined value.\",\n \"The third target throttle opening computing means calculates the target throttle opening in accordance with the second target throttle opening and makes it possible to adjust the actual air flow rate to the target air flow rate.\",\n \"To achieve another object, the internal combustion engine controller of the present invention uses the target throttle opening computing means, which comprises a computation permission means for permitting the second target throttle opening computing means to compute the target throttle opening in accordance with the operating conditions.\",\n \"The computation permission means inhibits the second target throttle opening computing means from computing the target throttle opening if a target exhaust gas re-circulation rate is not smaller than a predetermined value.\",\n \"The third target throttle opening computing means calculates the target throttle opening in accordance with the first target opening and makes it possible to adjust the actual air flow rate to the target air flow rate.\",\n \"As indicated in the configuration described above, the internal combustion engine controller of the present invention comprises a second computation permission means, which permits the second target throttle opening computing means to compute a throttle opening.\",\n \"Therefore, if, for instance, the EGR rate is as specified or the air flow sensor is normal, the second target throttle opening computing means can be permitted to computer the target throttle opening.\",\n \"Thus, it is possible to compute the target throttle opening that is responsive to changes in the internal combustion engine environment.\",\n \"To achieve another object, the internal combustion engine controller of the present invention uses the target throttle opening computing means to calculate the third target throttle opening from the first target throttle opening, which is computed in accordance with the actual air flow rate even when the relationship between the second target throttle opening and actual air flow rate significantly changes, and makes it possible to adjust the actual air flow rate for the target air flow rate.\",\n \"As indicated in the configuration described above, the internal combustion engine controller of the present invention exercises feedback control to provide computation control so as to adjust the actual air flow rate for the target air flow rate and calculate the target throttle opening even when the relationship between the target throttle opening computed by feed-forward control and the actual air flow rate significantly changes due to an air density change, foreign matter inclusion in an intake pipe, or other environmental change.\",\n \"Therefore, the air flow rate can be accurately controlled in response to the above environmental changes.\",\n \"To achieve another object, the internal combustion engine controller of the present invention uses the target throttle opening computing means, which comprises a target air flow rate correction means for correcting the target air flow rate.\",\n \"The first target throttle opening computing means computes the first target throttle opening in accordance with the difference between the target air flow rate calculated by the target air flow rate correction means and the actual air flow rate.\",\n \"The target air flow rate correction means represents a response characteristic between the second target throttle opening and actual air flow rate.\",\n \"As indicated by the configuration described above, the internal combustion engine controller of the present invention comprises the target air flow rate correction means.\",\n \"Therefore, it can accurately compute a target air flow rate while improving the transmission characteristic by compensating for a time delay involved in intake air flow from the throttle to a cylinder.\",\n \"To achieve another object, the internal combustion engine controller of the present invention uses the second target throttle opening computing means to compute the second target throttle opening in accordance with the internal combustion engine's rotating speed and the target air flow rate.\",\n \"As indicated by the configuration described above, the internal combustion engine controller of the present invention can exercise feed-forward control to compute the second target throttle opening, for instance, from a map depicting the relationship between the internal combustion engine's rotating speed and the target air flow rate.\",\n \"To achieve another object, the internal combustion engine controller of the present invention comprises means for computing the target exhaust gas re-circulation rate.\",\n \"Further, the second target throttle opening computing means computes the second target throttle opening in accordance with the internal combustion engine's rotating speed, the target air flow rate, and the target exhaust gas re-circulation rate.\",\n \"To achieve another object, the internal combustion engine controller of the present invention uses the target air flow rate computing means to compute the target air flow rate in accordance with the target air-fuel ratio and target fuel injection quantity.\",\n \"As indicated by the configuration described above, the internal combustion engine controller of the present invention can correct the second target throttle opening in accordance with the exhaust gas re-circulation rate when the EGR gas recirculates.\",\n \"More specifically, it can determine the target EGR rate from, for instance, a map depicting the relationship between a target combustion pressure torque and internal combustion engine's rotating speed, determine a throttle opening correction factor from, for instance, a map depicting the relationship between the target EGR rate and target air flow rate, and correct the second target throttle opening.\",\n \"To achieve another object, the internal combustion engine controller of the present invention comprises a detection means for detecting the internal combustion engine's exhaust gas components directly or indirectly, and a correction amount computing means for computing the correction amount for the target air flow rate in accordance with the exhaust gas components detected by the detection means.\",\n \"As indicated by the configuration described above, the internal combustion engine controller of the present invention corrects the target air flow rate with a correction amount (manipulated variable) that is used to compute (by means of air-fuel ratio feedback control) the correction amount for the target air flow rate in accordance with a signal from an A/F sensor or other exhaust gas component detection means.\",\n \"It is therefore possible to calculate a target air flow rate with increased accuracy and compute a target throttle opening.\",\n \"To achieve another object, the internal combustion engine controller of the present invention uses the second target throttle opening computing means to compute the second target throttle opening in accordance with an inverse model for the response between an in-cylinder target air flow rate and the throttle.\",\n \"To achieve still another object, the internal combustion engine controller of the present invention uses an in-cylinder air flow rate value, which is detected directly or indirectly, as an actual air flow rate value.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS FIG.\",\n \"1 illustrates the overall configuration of an internal combustion engine that comprises an internal combustion engine controller according to the present invention; FIG.\",\n \"2 illustrates the internal configuration of an internal combustion engine controller shown in FIG.\",\n \"1; FIG.\",\n \"3 is a control block diagram that illustrates a first embodiment of an internal combustion engine controller according to the present invention; FIG.\",\n \"4 is a block diagram that illustrates a target torque computing section shown in the control block diagram in FIG.\",\n \"3; FIG.\",\n \"5 is a block diagram that illustrates a fuel injection quantity computing section shown in the control block diagram in FIG.\",\n \"3; FIG.\",\n \"6 is a block diagram that illustrates a fuel injection quantity correction section shown in the control block diagram in FIG.\",\n \"3; FIG.\",\n \"7 is a block diagram that illustrates a target equivalence ratio computing section shown in the control block diagram in FIG.\",\n \"3; FIG.\",\n \"8 is a block diagram that illustrates a target air flow rate computing section shown in the control block diagram in FIG.\",\n \"3; FIG.\",\n \"9 is a block diagram that illustrates an actual air flow rate computing section shown in the control block diagram in FIG.\",\n \"3; FIG.\",\n \"10 is a block diagram that illustrates a target throttle opening computing section shown in the control block diagram in FIG.\",\n \"3; FIG.\",\n \"11 is a block diagram that illustrates a throttle opening control section for the target throttle opening computing section shown in FIG.\",\n \"10; FIG.\",\n \"12 is a control block diagram that illustrates an internal combustion engine controller according to a second embodiment of the present invention; FIG.\",\n \"13 is a control block diagram that illustrates a throttle opening computing section shown in the control block diagram in FIG.\",\n \"12; FIG.\",\n \"14 is a control block diagram that illustrates a target throttle opening computing section of an internal combustion engine controller according to a third embodiment of the present invention; FIG.\",\n \"15 is a control block diagram that illustrates a feedback control target value filter for the target throttle opening computing section shown in FIG.\",\n \"14; FIG.\",\n \"16 is a control block diagram that illustrates a target throttle opening computing section of an internal combustion engine controller according to a fourth embodiment of the present invention; FIG.\",\n \"17 is a block diagram that illustrates a feedback control permission judgment section for the target throttle opening computing section shown in FIG.\",\n \"16; FIG.\",\n \"18 is a control block diagram that illustrates a target throttle opening computing section of an internal combustion engine controller according to a fifth embodiment of the present invention; FIG.\",\n \"19 is a control block diagram that illustrates a feed-forward control permission judgment section for the target throttle opening computing section shown in FIG.\",\n \"18; FIG.\",\n \"20 is a control block diagram that illustrates a target throttle opening computing section of an internal combustion engine controller according to a sixth embodiment of the present invention; FIG.\",\n \"21 is a control block diagram that illustrates an actual air flow rate computing section of an internal combustion engine controller according to a seventh embodiment of the present invention; FIG.\",\n \"22 is a control block diagram that illustrates a target throttle opening computing section of an internal combustion engine controller according to a seventh embodiment of the present invention; FIG.\",\n \"23 is a control block diagram illustrating an inverse model for feed-forward control, which is within the target throttle opening computing section shown in FIG.\",\n \"22; FIG.\",\n \"24 is a control block diagram that illustrates an internal combustion engine controller according to an eighth embodiment of the present invention; FIG.\",\n \"25 is a control block diagram that illustrates an air-fuel ratio feedback control section shown in FIG.\",\n \"24; FIG.\",\n \"26 is a control block diagram that illustrates a target air flow rate computing section shown in FIG.\",\n \"24; FIG.\",\n \"27 is a block diagram that illustrates a conventional air-based torque-on-demand control; FIG.\",\n \"28 is a block diagram that illustrates a conventional fuel-based torque-on-demand control; FIG.\",\n \"29 shows in-cylinder air flow rate transient characteristic curves that are prevalent when an internal combustion engine's target air flow rate changes; FIG.\",\n \"30 shows HC, CO, and NOx purification characteristic curves of an internal combustion engine's three-way catalyst; and FIG.\",\n \"30 shows a correlation between an internal combustion engine's air-fuel ratio and internal combustion engine stability.\",\n \"DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Embodiments of an internal combustion engine controller according to the present invention will now be described in detail with reference to the accompanying drawings.\",\n \"FIG.\",\n \"1 illustrates an overall internal combustion engine system that is common to all embodiments to which an internal combustion engine controller according to the present invention is applied.\",\n \"An internal combustion engine 20 comprises a multi-cylinder, in-cylinder fuel injection internal combustion engine.\",\n \"Its intake system is such that the air coming from the outside passes through an air cleaner 1, an intake manifold 4, and a collector 5 in order named, and flows into each cylinder 9.The amount of air inflow is regulated by an electronic throttle 3.A spark plug 8 and a fuel injection valve 7 are mounted on each cylinder 9.Further, each cylinder 9 is provided with an intake valve 27 and an exhaust valve 28.The employed exhaust system is such that an exhaust manifold 10 is connected to each cylinder 10.A three-way catalyst 11 is mounted on the exhaust manifold 10.Further, an A/F sensor 12 is mounted between each cylinder 9 and the three-way catalyst 11.An exhaust gas re-circulation path (EGR path) 18 is furnished to establish communication between the intake manifold 4 and exhaust manifold 10 by bypassing each cylinder 9.The EGR path is provided with an EGR valve 19.The intake manifold 4 for the intake system is provided with an air flow sensor 2.The air flow sensor detects the amount of air inflow.\",\n \"A crank angle sensor 15 outputs a signal each time a crankshaft rotates one degree.\",\n \"A throttle opening sensor 17, which is mounted on an electronic throttle 3, detects the opening of the electronic throttle 3.A water temperature sensor 14 detects a cooling water temperature of the internal combustion engine 20.An accelerator opening sensor 13 detects the depression amount of an accelerator 6 for the purpose of detecting a torque demanded by the driver.\",\n \"The signals output from the accelerator opening sensor 13, air flow sensor 2, throttle opening sensor 17, crank angle sensor 15, and water temperature sensor 14 are transmitted to a control unit (controller) 50.The information about the operating status of the internal combustion engine 20 is derived from the signals output from these sensors to optimally compute major manipulated variables of the internal combustion engine, such as the intake air volume, fuel injection quantity, and ignition timing.\",\n \"The fuel injection quantity computed within the control unit 50 is converted to a valve open pulse signal and forwarded to the fuel injection valve 7.Further, the control unit 50 computes an ignition time as specified.\",\n \"To ensure that ignition takes place at the computed ignition time, the control unit 50 outputs a drive signal to the spark plug 8.The intake air delivered from the intake system is regulated by the electronic throttle 3, mixed with a re-circulated exhaust gas that is regulated by the EGR valve 19, and admitted into the cylinder 9 via the intake valve 27.The fuel injected from the fuel injection valve 7 into the cylinder (combustion chamber) 9 is mixed with an air inflow from the intake manifold 4 to form an air-fuel mixture.\",\n \"The air-fuel mixture is combustively burned by a spark that is generated from the spark plug 8 at the computed ignition time.\",\n \"The resulting combustion pressure pushes a piston 29 downward so as to generate motive power for the internal combustion engine 20.The exhaust gas generated upon combustion goes into the three-way catalyst 11 via the exhaust manifold 10.All exhaust gas components (HC, CO, and NOx) are then purified in the three-way catalyst 11 and discharged again to the outside.\",\n \"The amount of exhaust gas re-circulated to the intake side via an exhaust gas re-circulation pipe 18 is regulated by the EGR valve 19.The A/F sensor 12 has a linear output characteristic in relation to the concentration of oxygen contained in the exhaust gas.\",\n \"The relationship between the concentration of oxygen in the exhaust gas and the air-fuel ratio is substantially linear.\",\n \"Therefore, the A/F sensor 12, which detects the concentration of oxygen, can determine the air-fuel ratio of the internal combustion engine 20.The control unit 50 is configured so as to use the signal from the A/F sensor 12 to calculate the upstream air-fuel ratio of the three-way catalyst 11, and exercise feedback control to successively correct the fuel injection quantity or air flow rate for the purpose of ensuring that the air-fuel ratio of an air-fuel mixture in the cylinder 9 for the internal combustion engine 20 agrees with a target air-fuel ratio.\",\n \"FIG.\",\n \"2 illustrates the internal configuration of the control unit (ECU) 50 for the internal combustion engine 20 shown in FIG.\",\n \"1.The values output from the A/F sensor 12, throttle opening sensor 17, air flow sensor 2, internal combustion engine rotating speed sensor 15, and water temperature sensor 14 enter the ECU 50.The output values are then subjected to noise elimination and other signal processes in an input circuit 54, and forwarded to an input/output port 55.The value of the input/output port 55 is stored in a RAM 53 and subjected to arithmetic processing by a CPU 51.A control program containing the description of arithmetic processing is stored in a ROM 52 beforehand.\",\n \"The values representing the amounts of individual actuator operations, which are computed by the control program, are stored in the RAM 53 and then forwarded to the input/output port 55.An ON/OFF signal is set as the operation signal for the spark plug 8, which is used for ignition and combustion.\",\n \"This signal is ON when the primary coil in an ignition output circuit 56 is energized and OFF when the same circuit is de-energized.\",\n \"Ignition takes place when the signal status changes from ON to OFF.\",\n \"A spark plug signal, which is set for the input/output port 55, is amplified to energy adequate for combustion and then supplied to the spark plug 8.As a drive signal for the fuel injection valve 7, an ON/OFF signal is set.\",\n \"This signal turns ON to open the valve and turns OFF to close the valve.\",\n \"The signal is amplified to an adequate energy level that is required for operating the fuel injection valve drive circuit 57 to open the fuel injection valve 7, and then forwarded to the fuel injection valve 7.A drive signal for opening the electronic throttle 3 to a target degree of opening is delivered to the electronic throttle 3 via an electronic throttle drive circuit 58.The configuration common to all embodiments of an internal combustion engine controller according to the present invention has been described.\",\n \"From now on, individual embodiments will be described.\",\n \"First Embodiment The control program to be written into the ROM 52 of the EPU 50 will now be described.\",\n \"FIG.\",\n \"3 is a control block diagram illustrating the overall control operation that the EPU 50 shown in FIG.\",\n \"2 performs in accordance with a first embodiment.\",\n \"The major portion of a fuel-based, torque-on-demand control is shown in FIG.\",\n \"3.This control comprises a target torque computing section 61, a fuel injection quantity computing section 62, a fuel injection quantity correction section 63, a target equivalence ratio computing section 64, a target air flow rate computing section 65, an actual air flow rate computing section 66, a target throttle opening computing section 67, and a throttle opening control section 68.The target torque computing section 61 computes a target torque TgTc from an accelerator opening Apo and an internal computer combustion engine rotating speed Ne.\",\n \"The fuel injection quantity computing section 62 operates based on the target torque TgTc to compute a fuel injection quantity TIO that provides the target torque TgTc.\",\n \"The fuel injection quantity correction section 63 provides phase compensation so that the fuel injection quantity TIO matches the phase of the air in the cylinder 9, and then computes a corrected fuel injection quantity TI.\",\n \"The target equivalence ratio computing section 64 computes a target equivalence ratio TgFbya from the target torque TgTc and internal combustion engine rotating speed Ne.\",\n \"For the sake of convenience, the ratio between fuel and air is handled in the form of an equivalence ratio.\",\n \"It goes without saying that the air-fuel ratio may be used as is.\",\n \"It should be noted that the target equivalence ratio computing section 64 also chooses between homogeneous combustion and stratified combustion.\",\n \"The target air flow rate computing section 65 computes a target air flow rate TgTp from the fuel injection quantity TIO and target equivalence ratio TgFbya.\",\n \"Although this will be described in detail later, the amount of air inflow into a cylinder per cycle is standardized as a value of the target air flow rate TgTp for the sake of convenience.\",\n \"The actual air flow rate computing section 66 converts a mass flow rate Qa detected by the air flow sensor 2 into an actual air flow rate Tp, which is of the same dimension as the target air flow rate TgTP and flows into a cylinder per cycle, and then outputs the converted value.\",\n \"The target throttle opening computing section 67 computes a target throttle opening TgTvo in accordance with the target air flow rate TgTp and actual air flow rate Tp.\",\n \"The throttle opening computing section 68 computes a throttle manipulated variable Tduty from the target throttle opening TgTvo, the actual opening of the electronic throttle 3, and the value Tvo.\",\n \"The throttle manipulated variable Tduty represents the duty ratio of a PWM signal that enters a drive circuit 58, which controls a throttle motor drive current.\",\n \"The control computing sections and correction sections of the first embodiment of the control block described above will now be described in detail with reference to FIGS.\",\n \"4 to 12.1.Target Torque Computing Section FIG.\",\n \"4 illustrates the target torque computing section 61.The value TgTc represents a torque that is equivalent to a target combustion pressure.\",\n \"The value TgTa represents a torque required by the accelerator.\",\n \"The value TgTl represents an air flow rate that is required for idling speed maintenance.\",\n \"Since an off-idle state provides torque control and an idle state provides output control, it is assumed that the manipulated variable of an idling control means 61a is a theoretical air-fuel ratio, which is proportional to the output.\",\n \"Gain is provided for use in output-to-torque dimensional conversion.\",\n \"Further, when the air-conditioner (AC) is ON, the corresponding load is corrected.\",\n \"If any electrical load exists, the corresponding load is corrected.\",\n \"For the air flow rate required for idling speed maintenance TgTl, a lower-limit value is defined as a BCV portion for assuring an adequate air flow rate for avoiding a reverse oil flow into the cylinder 6.An idle feedback control means 61a2 functions during an idle state only in order to correct feed-forward-related error.\",\n \"It is assumed that the controlled variable for the idle feedback control means 61a2 is the rotating speed Ne, and that the manipulated variable is an air flow rate.\",\n \"The algorithm for the idle feedback control means 61a2 is not specified herein.\",\n \"However, it may be, for instance, a PID control algorithm.\",\n \"An idling judgment section 61b concludes that an idle state exists when the accelerator opening Apo is smaller that specified.\",\n \"It is desirable that the TblTgTa and TblTgTf settings be determined according to actual device data.\",\n \"2.Fuel Injection Quantity Computing Section FIG.\",\n \"5 illustrates the fuel injection quantity computing section 62, which converts a target combustion pressure torque TgTc to a fuel injection quantity TIO for control purposes.\",\n \"The fuel injection quantity TIO is proportional to torque because it represents a fuel injection quantity per cylinder per cycle.\",\n \"This proportional relationship is used to convert the target combustion pressure torque TgTc to the fuel injection quantity TIO.\",\n \"A gain may be used, but table conversion may be effected in consideration of some possible error.\",\n \"It is desirable that the setting be determined according to actual device data.\",\n \"3.Fuel Injection Quantity Correction Section FIG.\",\n \"6 illustrates the fuel injection quantity correction section 63, which provides correction to adjust the fuel injection quantity TIO for the phase of the air in the cylinder 9.The air transmission characteristic between the throttle 3 and cylinder 9 is approximated with wasted time plus first-order lag.\",\n \"It is desirable that the settings for parameter n1, which represents wasted time, and parameter Kair, which corresponds to a time constant for a first-order lag, be determined according to actual device data.\",\n \"Further, parameter n1 and parameter Kair may be varied depending on the operating conditions.\",\n \"4.Target Equivalence Ratio Computing Section FIG.\",\n \"7 illustrates the target equivalence ratio computing section 64, which determines the combustion status and computes the target equivalence ratio.\",\n \"Fpstratify is a stratified combustion permission flag.\",\n \"When Fpstratify=1, the injection time, ignition time, injection quantity, and air flow rate are regulated to provide stratified combustion.\",\n \"Injection time/ignition time determination is not described herein.\",\n \"The stratified combustion permission flag Fpstratify is 1 to permit stratified combustion if the conditions are satisfied by the water temperature Twn, accelerator opening Apo, and rotating speed Ne.\",\n \"When stratified combustion is permitted, the value for letting a stratified combustion target equivalence ratio map Mtgfba_s be referenced by the target combustion pressure torque TgTc and rotating speed Ne is the target equivalence ratio TgFbya.\",\n \"When TgFbya=0, homogeneous combustion takes place, and the value for letting a homogeneous combustion target equivalence ratio map Mtgfba be referenced by the target combustion pressure TgTc and rotating speed Ne is the target equivalence ratio TgFbya.\",\n \"It is desirable that the settings for the stratified combustion target equivalence ratio map Mtgfba_s and homogeneous combustion target equivalence ratio map Mtgfba be determined according to actual device data.\",\n \"5.Target Air Flow Rate Computing Section FIG.\",\n \"8 illustrates the target air flow rate computing section 65, which computes the target air flow rate TgTp.\",\n \"For the sake of convenience, the target air flow rate TgTp is computed as a standardized value representing the air flow rate that flows into a cylinder per cycle.\",\n \"As is obvious from FIG.\",\n \"8, the target air flow rate TgTp is calculated from the fuel injection quantity TIO and target equivalence ratio TgFbya.\",\n \"The equation used for this calculation is as follows: TgTp=TIO×(1/TgFbya) 6.Actual Air Flow Rate Computing Section FIG.\",\n \"9 illustrates the actual air flow rate computing section 66, which computes the actual air flow rate Tp.\",\n \"For the sake of convenience, the actual air flow rate Tp is computed as a standardized value representing the air flow rate that flows into a cylinder per cycle.\",\n \"The value Qa is an incoming air flow rate that is detected by the air flow sensor 2.The value K is determined so that the actual air flow rate Tp provides the fuel injection quantity prevalent at the theoretical air-fuel ratio.\",\n \"The value Cy1 denotes the number of cylinders for the internal combustion engine 20.7.Target Throttle Opening Computing Section FIG.\",\n \"10 illustrates the target throttle opening computing section 67, which determines the target throttle opening TgTVO from the target air flow rate TgTp, actual air flow rate Tp, and rotating speed Ne.\",\n \"This control block can be divided into a section 67b (second target throttle opening computing means) for exercising feed-forward control to determine the target throttle opening TgTVOFF from the target air flow rate TgTp and rotating speed Ne and a feedback control section 67a (first target throttle opening computing means) for exercising feedback control to determine the target throttle opening TgTVOFB from the target air flow rate TgTp and actual air flow rate Tp.\",\n \"It is assumed, as shown in FIG.\",\n \"10, that the feed-forward control section 67b determines the target throttle opening TgTVOFF by referencing a map.\",\n \"It is desirable that the map setting be determined according to actual device data.\",\n \"Feedback control is exercised to provide PID control.\",\n \"Each gain is given as a magnitude of deviation between the target air flow rate TgTp and actual air flow rate TpTg.\",\n \"However, it is desirable that the setting be determined according to actual device data.\",\n \"For the D portion, an LPF (low-pass filter) is provided for high-frequency noise elimination.\",\n \"The target throttle opening TgTVOFF that is computed by the second section 67b of feed-forward control, and the target throttle opening TgTVOFB that is determined by the first section 67a of feedback control are output to the third control section 67c (third throttle opening computing means).\",\n \"The third control section 67c then computes the sum of the target throttle opening TgTVOFF and target throttle opening TgTVOFB and uses the computed sum as the final target throttle opening TgTVO.\",\n \"8.Throttle Opening Control Section FIG.\",\n \"11 illustrates the throttle opening control section 68, which computes the throttle drive manipulated variable Tduty from the target throttle opening TgTVO and actual throttle opening Tvo.\",\n \"As described earlier, the throttle drive manipulated variable Tduty denotes the duty ratio of a PWM signal that enters a drive circuit 58, which controls a throttle motor drive current.\",\n \"It is assumed that the throttle drive manipulated variable Tduty is determined by means of PID control.\",\n \"Although no details will be given, it is desirable that each PID control gain be tuned with an actual device for optimum results.\",\n \"Second Embodiment The control block of an internal combustion engine controller according to a second embodiment of the present invention will now be described in detail.\",\n \"FIG.\",\n \"12 is a control block diagram of the second embodiment that illustrates the overall control of the EPU 50 shown in FIG.\",\n \"2.The major portion of a fuel-based, torque-on-demand control is shown in FIG.\",\n \"12.The target torque computing section 61, fuel injection quantity computing section 62, fuel injection quantity correction section 63, target equivalence ratio computing section 64, target air flow rate computing section 65, actual air flow rate computing section 66, and target throttle opening computing section 67 within the control block of the second embodiment are the same as their counterparts described in Sections 1 to 7, respectively, under the first embodiment and will not be described again.\",\n \"8.Throttle Opening Control Section FIG.\",\n \"13 illustrates the throttle opening control section 68.As indicated in FIG.\",\n \"13, feed-forward control is exercised to determine the throttle drive manipulated variable Tduty from the target throttle opening TgTVO only.\",\n \"The feed-forward control portion may be a gain only.\",\n \"In such an instance, however, the manipulated variable (Tduty) for throttle opening control is computed directly from a target air flow rate TgTp without computing a target throttle opening.\",\n \"Third Embodiment The control block of an internal combustion engine controller according to a third embodiment of the present invention will now be described in detail.\",\n \"The control block of the third embodiment differs from that of the first embodiment, which is shown in FIG.\",\n \"3, in that the target air flow rate TgTp for use in feedback control is filtered in relation to a feed-forward control target value in the target throttle opening computing section 67.The filter characteristic will be described in detail later.\",\n \"However, it is a response characteristic between the target throttle opening and in-cylinder actual air flow rate that is computed by means of feed-forward control in the target air flow rate computing section 65.The purpose is to exercise feedback control to compensate for the steady-state deviation.\",\n \"The target torque computing section 61, fuel injection quantity computing section 62, fuel injection quantity correction section 63, target equivalence ratio computing section 64, target air flow rate computing section 65, actual air flow rate computing section 66, and throttle opening control section within the control block of the third embodiment are the same as their counterparts described in Sections 1 to 6 and 8, respectively, under the first embodiment and will not be described again.\",\n \"7.Target Throttle Opening Computing Section As shown in FIG.\",\n \"14, the target air flow rate TgTp for feedback control is a value that is obtained by filtering the target air flow rate TgTp with a filter 67d.\",\n \"As indicated in FIG.\",\n \"15, the specifications for the filter 67c are such that the air transmission characteristic between the throttle 3 and cylinder 9 is approximated with wasted time plus first-order lag.\",\n \"It is desirable that the settings for parameter n2, which represents wasted time, and parameter Kair2, which corresponds to a time constant for a first-order lag, be determined according to actual device data.\",\n \"Further, the parameter n2 and the parameter Kair2 may be varied depending on the operating conditions.\",\n \"Fourth Embodiment The control block of an internal combustion engine controller according to a fourth embodiment of the present invention will now be described in detail.\",\n \"The control block of the fourth embodiment differs from that of the first embodiment, which is shown in FIG.\",\n \"3, in that the feedback control's computation of target throttle opening TgTVO can be permitted depending on the operating conditions in the target throttle opening computing section 67.The target torque computing section 61, fuel injection quantity computing section 62, fuel injection quantity correction section 63, target equivalence ratio computing section 64, target air flow rate computing section 65, actual air flow rate computing section 66, and throttle opening control section within the control block of the fourth embodiment are the same as their counterparts described in Sections 1 to 6 and 8, respectively, under the first embodiment and will not be described again.\",\n \"7.Target Throttle Opening Computing Section When the feedback control computation permission flag FPFB is 1 in the feedback control permission judgment section 67e, a selector section 67f effects a changeover, as shown in FIG.\",\n \"16, so that the target throttle opening TgTVOFB is determined by means of feedback control.\",\n \"When the feedback control computation permission flag FPFB is 0, on the other hand, the target throttle opening TgTVOFB is set to 0 without exercising feedback control to perform computations.\",\n \"As indicated in FIG.\",\n \"17, the feedback control computation permission flag FPFB is established when the accelerator opening Apo and the time elapse after internal combustion engine startup Tafst are as specified and the air flow sensor and throttle are both normal.\",\n \"The diagnostic check methods for the air flow sensor 2 and throttle 3 are not particularly described herein; however, various practical methods are available.\",\n \"Fifth Embodiment The control block of an internal combustion engine controller according to a fifth embodiment of the present invention will now be described in detail.\",\n \"The control block of the fifth embodiment differs from that of the first embodiment, which is shown in FIG.\",\n \"3, in that the feed-forward control's computation of target throttle opening TgTPVOFF can be permitted depending on the operating conditions in the target throttle opening computing section 67.The target torque computing section 61, fuel injection quantity computing section 62, fuel injection quantity correction section 63, target equivalence ratio computing section 64, target air flow rate computing section 65, actual air flow rate computing section 66, and throttle opening control section within the control block of the fifth embodiment are the same as their counterparts described in Sections 1 to 6 and 8, respectively, under the first embodiment and will not be described again.\",\n \"7.Target Throttle Opening Computing Section When the feed-forward control computation permission flag FPFB is 1 in the feed-forward control permission judgment section 67g, the target throttle opening TgTVOFF is determined by means of feed-forward control as shown in FIG.\",\n \"18.When the feed-forward control computation permission flag FPFF is 0, on the other hand, the target throttle opening TgTVOFF is set to 0 without exercising feed-forward control to perform computations.\",\n \"As indicated in FIG.\",\n \"19, the feed-forward control computation permission flag FPFF is established when the target EGR rate TgEgr is as specified and the air flow sensor is normal.\",\n \"The diagnostic check method for the air flow sensor is not particularly described herein.\",\n \"Sixth Embodiment The control block of an internal combustion engine controller according to a sixth embodiment of the present invention will now be described in detail.\",\n \"The control block of the sixth embodiment differs from that of the first embodiment, which is shown in FIG.\",\n \"3, in that feed-forward control is exercised in the target throttle opening computing section 67 to compute the target throttle opening TgTVOFF while considering the exhaust gas re-circulation rate.\",\n \"The target torque computing section 61, fuel injection quantity computing section 62, fuel injection quantity correction section 63, target equivalence ratio computing section 64, target air flow rate computing section 65, actual air flow rate computing section 66, and throttle opening control section within the control block of the sixth embodiment are the same as their counterparts described in Sections 1 to 6 and 8, respectively, under the first embodiment and will not be described again.\",\n \"7.Target Throttle Opening Computing Section As indicated in FIG.\",\n \"20, the target throttle opening TgTVOFF, which is determined by means of feed-forward control, is calculated according to the following equation: TgTVOFF=TgTVOFF0×EgrHos where EgrHOS is a throttle opening correction factor that is used in the event of EGR, and the target throttle opening TgTVOFF0 is determined in the same manner as with the first embodiment.\",\n \"As indicated in FIG.\",\n \"20, a map is referenced in accordance with the target combustion pressure torque TgTc and rotating speed Ne to determine the target EGR rate TgEgr.\",\n \"Further, a map is referenced in accordance with the target EGR rate TgEgr and target air flow rate TgTP to determine the throttle opening correction factor EgrHos.\",\n \"It is desirable that the settings for the maps be determined according to actual device data.\",\n \"Seventh Embodiment The control block of an internal combustion engine controller according to a seventh embodiment of the present invention will now be described in detail.\",\n \"The control block of the seventh embodiment differs from that of the first embodiment, which is shown in FIG.\",\n \"3, in that feed-forward control is exercised in the target throttle opening computing section 67 to compute the target throttle opening TgTVOFF in accordance with an inverse model for the response characteristic between the air flow rate in the cylinder 9 and throttle 3.Further, the air flow rate in the cylinder 9 is used as the actual air flow rate Tp for feedback control.\",\n \"The target torque computing section 61, fuel injection quantity computing section 62, fuel injection quantity correction section 63, target equivalence ratio computing section 64, target air flow rate computing section 65, and throttle opening control section within the control block of the seventh embodiment are the same as their counterparts described in Sections 1 to 5 and 8, respectively, under the first embodiment and will not be described again.\",\n \"6.Actual Air Flow Rate Computing Section As indicated in FIG.\",\n \"21, the air flow rate Tp1 flowing into a cylinder per cycle is subjected to the air transmission characteristic prevalent between the throttle and cylinder.\",\n \"The transmission characteristic is approximated with wasted time plus first-order lag.\",\n \"It is desirable that the settings for parameter n3, which represents wasted time, and parameter Kair3, which corresponds to a time constant for a first-order lag, be determined according to actual device data.\",\n \"Further, the parameter n3 and the parameter Kair3 may be varied depending on the operating conditions.\",\n \"7.Target Throttle Opening Computing Section As indicated in FIG.\",\n \"22, an inverse filter (inverse model) 67k is used to determine the target throttle opening TgTVOFF for feed-forward control.\",\n \"Detailed specifications for the inverse filter 67k are shown in FIG.\",\n \"23.It is desirable that the settings for a1, b0, and b1 be determined according to actual device data.\",\n \"It is also conceivable that the settings may be varied depending on the operating conditions.\",\n \"Eighth Embodiment The control block of an internal combustion engine controller according to an eighth embodiment of the present invention will now be described in detail.\",\n \"FIG.\",\n \"24 is a control block diagram of the eighth embodiment that illustrates the overall control of the EPU 50 shown in FIG.\",\n \"2.The major portion of a fuel-based, torque-on-demand control is shown in FIG.\",\n \"24.The control block of the eighth embodiment differs from that of the first embodiment, which is shown in FIG.\",\n \"3, in that an air-fuel ratio feedback control section 69 is added.\",\n \"The air-fuel ratio feedback control section 69 provides a manipulated variable Alpha, which is used to correct the target air flow rate TgTP that is computed by the target air flow rate computing section 65.The above functionality is added so as to correct the target air flow rate TgTP in accordance with the value output from the A/F sensor 12, which is mounted on the exhaust manifold 10.The target torque computing section 61, fuel injection quantity computing section 62, fuel injection quantity correction section 63, target equivalence ratio computing section 64, actual air flow rate computing section 66, target throttle opening computing section 67, and throttle opening control section 68 within the control block of the eighth embodiment are the same as their counterparts described in Sections 1 to 4 and 7 to 9, respectively, under the first embodiment and will not be described again.\",\n \"5.Air-Fuel Ratio Feedback Control Section This section exercises feedback control in accordance with the equivalence ratio Rfbya, which is detected by the A/F sensor 12, so that the internal combustion engine's equivalence ratio agrees with the target equivalence ratio TgFbya under arbitrary operating conditions.\",\n \"More specifically, PI control is exercised as indicated in FIG.\",\n \"25 to compute the air-fuel ratio correction term Alpha from the deviation between the target equivalence ratio TgFbya and the equivalence ratio Rfbya detected by the A/F sensor.\",\n \"6.Target Air Flow Rate Computing Section As shown in FIG.\",\n \"26, the target air flow rate TgTp is computed from the target equivalence ratio TgFbya and the equivalence ratio Rfbya detected by the A/F sensor.\",\n \"For computation, the following equation is used: TgTp=TIO×(1/(TgFbya×Alpha)) Although the present invention has been described in detail with reference to the first to eighth embodiments thereof, it will be readily apparent to those skilled in the art that the present invention is not limited to those preferred embodiments, and that various design changes may be made therein without departing from the spirit and scope of the appended claims.\",\n \"In the target throttle opening computing section, for instance, the target throttle opening computed by the second throttle opening computing means and the target throttle opening computed by the first throttle opening computing means are output to the third throttle opening computing means, which computes the sum of the first and second target throttle opening values and uses the computed sum as the final target throttle opening.\",\n \"However, the third throttle opening computing means can alternatively compute the final target throttle opening while considering not only the sum but also the product or other control element.\",\n \"In the foregoing embodiments, an air flow sensor is adopted as means for measuring the air flow rate and used to measure a mass flow rate.\",\n \"In an alternative embodiment, however, a pressure sensor for measuring the intake pipe internal pressure may also be used.\",\n \"Industrial Applicability As described above, the internal combustion engine controller according to the present invention comprises a first throttle opening computing means, which computes a throttle opening in accordance with a value detected by an air flow sensor, and a second throttle opening computing means, which computes a throttle opening without using a value detected by an air flow sensor.\",\n \"Therefore, when used in a fuel-based torque-on-demand control type internal combustion engine, the internal combustion engine controller according to the present invention can improve air flow rate response and convergence, reduce the exhaust gas emission, and provides improved torque response without being affected by various environmental changes or sensor, actuator, and other component variations.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473772"}}},{"rowIdx":752,"cells":{"text":{"kind":"list like","value":[["Stabilised interbody fusion system for vertebrae","The invention concerns a stabilised interbody fusion system for vertebrae, of the type comprising an interbody implant (4) designed to be inserted in the intervertebral space defined between two neighbouring vertebrae to be mutually secured, so as to restore the height and the angle of the lordosis of the vertebral segment defined by the two neighbouring vertebrae and a stabilizing plate (17) provided, at each of its ends, with at least a passage hole (18) for an anchoring screw, the plate (17) and the implant (4) being provided with mutual assembly means, such that after assembly, the stabilizing plate (17) extends on each side of the implant to enable the stabilizing plate to be anchored on the neighbouring vertebrae through the screws, characterised in that it comprises spacing means (30), interposed between the stabilizing plate (17) and the implant (4), to enable the stabilizing plate to be positioned at a specific distance relative to the implant."],["1-14.","(canceled) 15.Stabilized interbody fusion system for vertebrae according to the invention, of the type comprising: an interbody implant that will be inserted in the intervertebral space defined between two neighboring vertebrae to be mutually secured, in order to restore the height and the angle of the lordosis of the vertebral segment defined by two neighboring vertebrae; and a stabilizing plate equipped with at least one passage hole for an anchoring screw at each of its ends, the plate and the implant being provided with mutual assembly means such that after assembly, the stabilizing plate extends on each side of the implant to enable the stabilizing plate to be anchored on the neighboring vertebrae through the screws, wherein it comprises spacing means, interposed between the stabilizing plate and the implant, to enable the stabilizing plate to be positioned at a specific distance relative to the implant.","16.System according to claim 15, wherein the spacing means adjust the distance between the stabilizing plate and the implant.","17.System according to claim 15, wherein the spacing means are chosen from among a range of spacing means each with a determined length, each of which is different, to adjust the distance between the plate and the implant at a value that is determined by the length of the chosen spacing means.","18.System according to claim 15, wherein the assembly means are provided with support means for the plate allowing angular orientation of the plate relative to the implant.","19.System according to claim 16, wherein the assembly means are provided with support means for the plate allowing angular orientation of the plate relative to the implant.","20.System according to claim 17, wherein the assembly means are provided with support means for the plate allowing angular orientation of the plate relative to the implant.","21.System according to claim 15, wherein the spacing means comprises at least one spacer bushing.","22.System according to claim 16, wherein the spacing means comprises at least one spacer bushing.","23.System according to claim 17, wherein the spacing means comprises at least one spacer bushing.","24.System according to claim 18, wherein the spacing means comprises at least one spacer bushing.","25.System according to claim 21, wherein the spacer bushing is fixed to the stabilizing plate.","26.System according to claim 25, wherein the spacer bushing comprises assembly means on the implant, allowing adjustment of the distance between the stabilizing plate and the implant.","27.System according to claim 25, wherein the spacer bushing is put into place on an assembly screw forming part of the assembly means and is designed to pass through the plate through a passage hole bordered by a stop shoulder arranged to hold the head of the assembly screw and to cooperate with at least one threaded hole arranged in the implant.","28.System according to claim 27, wherein the threaded hole is formed on the anterior wall of the implant, this wall having a convex curvature in the transverse plane and extending by connecting walls with a convex curvature, to a posterior wall with a concave curvature to leave the vertebral canal free.","29.System according to claim 28, wherein each connecting wall is arranged to contain a threaded hole surrounded, like the threaded hole formed in the anterior wall, by an assembly groove designed to cooperate with two assembly dog pins provided on the head of a gripping instrument of the implant, also comprising a threaded rod designed to cooperate with a threaded hole in the implant.","30.System according to claim 28, wherein the posterior wall is fitted with a radio-opaque element opening up on the external face of the said wall, adapted so that the position of the implant within the intervertebral space along the sagittal plane can be detected.","31.System according to claim 15, wherein the stabilizing plane is provided with anti-expulsion means for the anchoring screws, installed on the plate through moving guide means and adapted to occupy a first position in which the passage cross-section of the holes for reception of the anchoring screws is left free, and a second position, in which the holes are partially closed off, in order to form a stop for the heads of screws.","32.A gripping instrument, comprising: a maneuvering tube, inside which a flexible threaded rod is fitted so as to cooperate with a threaded hole in the implant of claim 15, the tube further comprising an assembly head, fitted with two dog pins to cooperate with an assembly groove surrounding the threaded hole.","33.The gripping instrument according to claim 32, wherein the maneuvering tube on the implant has a curvature and is provided with a slide on which there is a bar, the free end of which is adapted to resist forces for placement of the implant and to ensure that the approach is respected."],["The present invention relates to interbody implants to be inserted in the intervertebral space defined between two neighbouring vertebrae after intervertebral disk excision, in order to restore the intervertebral space and to form bone fusion between the said neighbouring vertebrae.","More precisely, the domain of the invention is lumbar type interbody implants to restore the height and the angle of the lordosis of the vertebral segment defined by two neighbouring lumbar vertebrae to be mutually secured.","More precisely, the purpose of the invention is an interbody implant, called a secured implant, in other words provided with the means of anchoring the implant in its insertion site, to prevent it from becoming mobile.","There are several stabilised interbody fusion systems for vertebrae known in the state of the art.","For example, there is a known stabilised system comprising an interbody implant to be inserted in the intervertebral space defined between two neighbouring lumbar vertebrae.","According to one preferred embodiment, this implant is in the form of a cage comprising two sagittal walls connected to each other through an anterior transverse wall and a posterior transverse wall.","The walls delimit a volume between them opening up on each side of the transverse faces of the implant.","The open volume of the cage will receive a bone filling product called a bone graft, that will come into contact with the vertebral plates to facilitate bone fusion between the two vertebrae.","This type of stabilised system also comprises a stabilising plate extending from the anterior wall of the implant and on each side of the transverse faces.","This stabilising plate is provided with passage holes at each of its ends for screws that will be anchored in the vertebrae to be mutually secured.","A stabilising plate can be placed to reliably prevent migration of the implant.","However, the applicant has identified a difficulty in keeping the implant in its ideal position within the intervertebral space to enable it to be anchored on the vertebrae to be mutually assembled through the screws.","It must be borne in mind that, in different patients, firstly the anterior edges of the plates of the vertebra bodies have different profiles, and secondly implants are located at different distances from the anterior edges of the plates of the vertebra bodies, that causes displacement of the implant while the stabilising plate is anchored on the vertebrae.","The applicant expressed the need to have a stabilised interbody fusion system designed firstly so that the implant can be placed in the intervertebral space in an appropriate position to restore a physiological spinal curvature and lordosis, and secondly to hold the implant in this ideal correction position during anchorage of the stabilising plate on the vertebrae, regardless of the shape of the anterior edges of the plates of the vertebra bodies and the position of the implant relative to these anterior edges of the plates.","Therefore, the purpose of the invention is to satisfy this need by proposing a stabilised interbody fusion system capable of restoring the height and the angle of the lordosis of the vertebral segment by an implant, the function of which is secured by a stabilising plate fixed on the vertebrae.","In order to achieve this objective, the stabilised interbody fusion system for vertebrae according to the invention is of the type comprising: an interbody implant that will be inserted in the intervertebral space defined between two neighbouring vertebrae to be mutually secured, in order to restore the height and the angle of the lordosis of the vertebral segment defined by two neighbouring vertebrae, and a stabilising plate equipped with at least one passage hole for an anchoring screw at each of its ends, the plate and the implant being provided with mutual assembly means such that after assembly, the stabilising plate extends on each side of the implant to enable the stabilising plate to be anchored on the neighbouring vertebrae through the screws, The stabilised system comprises spacing means interposed between the stabilising plate and the implant, to enable the stabilising plate to be positioned at a specific distance relative to the implant.","Various other special features will become clear from the description given below with reference to the appended drawings that show example embodiments of the purpose of the invention as non-limitative examples.","FIG.","1 is an exploded perspective view showing the different constituents of the stabilised system according to the invention.","FIG.","2 is a sectional-elevation view of a stabilised system in the raised position.","FIG.","3 is an elevation view showing a stabilised system installed on two vertebrae.","FIG.","4 is a sectional-elevation view showing another variant embodiment of the stabilised system according to the invention.","FIG.","5 and 6 are perspective views showing characteristic details of a gripping instrument for an implant according to the invention.","FIG.","7 is a sectional-elevation view illustrating another variant embodiment of a stabilised system according to the invention.","As is clear particularly in FIG.","1 to 3, the invention relates to a stabilised interbody fusion system 1 for two neighbouring or nearby vertebrae 2, 3.The stabilised system 1 comprises an interbody implant 4 that will be inserted into the intervertebral space 5 delimited by the two vertebrae 2, 3.In one preferred embodiment of the invention as illustrated on the drawings, the implant 4 is made by a cage with a transverse wall 6 called the anterior wall, and a transverse wall 7 called the posterior wall, connected to each other by connecting walls 8, 9.The walls 6 to 9 of the cage are perpendicular to a transverse plane T that is perpendicular to a sagittal plane S passing through the anterior wall 6 and the posterior wall 7.In the transverse plane T, the anterior wall 6 has a convex curvature and extends on each side by the connecting walls 8, 9, which also have convex curvatures and are connected to each other by the posterior wall 7 with a concave curvature to leave the vertebral canal free.","The cage 1 thus has a general kidney shape.","As can be seen particularly in FIG.","2, the connecting walls 8, 9 in the sagittal plane S converge towards each other towards the posterior wall 7, such that the cage 1 has a tapered section in the sagittal plane S defining a lordosis correction angle.","The cage 1 comprises an interior volume 11 delimited by interior vertical faces of the walls 6 to 9 and will be filled by a bone filling product called a bone graft adapted to interbody fusion.","This volume 11 opens up on a first transverse face 12 called the upper face in the example illustrated, and a second transverse face 13 called the lower face.","These transverse faces 12, 13 are delimited by edges of the walls 6 to 9, preferably arranged to comprise notches 14 enabling the cage to be bonded to the supedjacent and subjacent vertebrae.","The notches 14 extend parallel to each other and from a frontal plane F perpendicular to the sagittal plane S and the transverse plane T. Preferably, the internal volume 11 of the cage comprises one or several connecting walls 15 with other walls.","In the example illustrated, the connecting wall 15 is in the general shape of a “Y” and extends between the connecting walls 8 and 9 and the anterior wall 6.The stabilised interbody fusion system 1 conform with the invention also comprises a stabilising plate 17 for the implant 4.This stabilising plate 17 is provided with at least one passage hole at each of its ends, and in the example illustrated with two passage holes 18 for the screws 19 that will be anchored on vertebrae 2 and 3 to be mutually secured.","According to one preferred characteristic embodiment, the stabilising plate 17 is provided with anti-expulsion means 20 for the anchoring screws 19.These anti-expulsion means 20 are composed of moving flaps installed on the stabilising plate so that they can be guided in rotation in order to occupy a first position in which the passage cross-section of the holes 18 is left free, and a second position in which this passage cross-section is closed off in order to form a stop for the heads of screws 19.The stabilising plate 17 and the implant 4 are provided with mutual assembly means 21.The stabilising plate 17 is adapted so that after assembly with the implant 4, it extends from each side of the transverse faces 12, 13 to the other, to anchor the screws 19 on the vertebrae 2, 3.According to one characteristic embodiment, the assembly means 21 consist of an assembly screw 22 that will cooperate with at least one tapped hole 23 formed in the implant 4.For example, a tapped hole 23 is arranged to pass through the anterior wall from one side to the other along an axis perpendicular to the anterior wall 6 and centred on the sagittal plane S of symmetry of the implant 4.Preferably, the implant 4 comprises a series of threaded holes 23 to enable assembly of the stabilising plate 17 in different positions with respect to the implant 4.Thus, a threaded hole 23 is formed in a connecting wall, for example 9, along an axis perpendicular to the axis of the threaded hole 23 formed in the anterior wall 6.At the positions defined by these two threaded holes 23, the stabilising plate 17 is free to occupy two positions perpendicular to each other.","Another threaded hole 23 may also be formed in the other connecting wall, 8, at an angle of 45° from the axis of the threaded hole 23 formed in the anterior wall 6.The assembly screw 22 passes through the stabilising plate 17 through a passage hole 24, bordered by a stop shoulder 25 formed by a dish 26 arranged to hold the head 221 of the assembly screw 22.According to one characteristic of the invention, the stabilised device 1 comprises spacing means 30 inserted between the stabilising plate 17 and the implant 4, to enable the stabilising plate 17 to be positioned at a specific distance relative to the implant 4.These spacing means 30 fix the stabilising plate 17 onto the vertebrae leaving the implant 4 in its position, taking account of the shape of the anterior edges B of the plates of vertebrae bodies 2, 3 or the setback position of the implant 4 relative to these anterior edges of the vertebrae.","In the example illustrated in FIG.","1 to 3, the spacing means 30 are composed of a spacer bushing 31 provided with a thread 32 cooperating with the thread of the assembly screw 22.The spacer bushing 31 is thus made in the form of a nut and its outside shape is prismatic and is adapted to cooperate with a part 33 for reception of the implant 4 in which the threaded hole 23 opens up.","Cooperation of the spacer bushing 31 with the reception part 33 blocks the bushing 31 in rotation when the assembly screw 22 is being screwed.","Preferably, the reception part 33 is made by an entry cone cooperating with a complementary part 34 presented by the bushing 31.Cooperation of the bushing 31 with the entry cone 33 is a means of adjusting the distance between the stabilising plate 17 and the implant 4.The stabilised system 1 according to the invention is used directly as a function of the description given above.","Before the implant 4 is inserted, the disc is excised by preparation of the vertebral plates.","The implant 4 is implanted in the intervertebral space 5 between two adjacent vertebrae 2 and 3 to be mutually secured.","The implant 4 is designed to restore the height and the angle of the lordosis of the vertebral segment defined by the two neighbouring vertebrae 2, 3.The stabilising plate 17, fitted with its assembly screw 22 provided with its spacer bushing 31, will be installed on the implant 4.The screw 22 is then screwed into a threaded hole 23 of the implant.","The screw 22 is then screwed in until it is fixed in place by the spacer bushing 31 being stopped between the plate 17 and the implant 4.The screws 19 are then anchored in the vertebrae 2, 3.Obviously, the length of the spacer bushing 31 is adapted to allow the implant 4 to remain in its implantation site, while allowing the stabilising plate 17 to come into contact with the vertebrae.","The stabilising plate 17 and the implant 4 are separated by a fixed measurement determined by the spacer bushing 31.Preferably, a range of spacer bushings 31 is available.","Each of these spacer bushings 31 has a determined length, each of which is different, to adjust the distance between the implant 4 and the stabilising plate 17.Note that the implant 4 is preferably provided with at least one radio-opaque element 35 (FIGS.","1 and 2) adapted so that the position of the implant 4 within the intervertebral space 5 along the sagittal plane S can be detected on an X-ray.","In the example illustrated, the radio-opaque element 35 is installed in the posterior wall 7 extending in the transverse plane and opening up on the external face and the internal face of the posterior wall 7.According to one example embodiment illustrated in FIG.","4, the assembly means 21 are provided with support means 36 for the stabilising plate 17 allowing angular orientation of the stabilising plate relative to the implant 4.For example, these support means 36 may be made by a ball joint type articulation formed between the head 221 of the assembly screw 22 and the dish 26 in the plate 17.In this variant embodiment illustrated, the screw 22 is assembled to the stabilising plate 17 using a ring 38 fixed on the plate 17 to trap the head 221 of the screw 22, while allowing the screw 22 free to rotate about its own axis.","According to one preferred variant embodiment, each threaded hole 23 of the implant is surrounded by an assembly groove 40, inclined relative to the sagittal plane and designed to cooperate with a gripping or manipulation instrument 44 for use on the implant 4.As can be seen in FIGS.","5 and 6, the manipulation instrument 44 comprises an assembly head 45 prolonged by a manoeuvring tube 46 inside which a flexible threaded rod 47 passing through the head 45 is installed, so as to cooperate with a threaded hole 23 in the implant.","The assembly head 45 is provided with two dog pins 49 that will be inserted in the groove 40 on each side of the threaded hole 23.The instrument 44 thus enables complete gripping and blocking of the implant 4 with respect to the assembly head 45.According to one preferred variant embodiment, the manoeuvring tube 46 has a curvature and is provided with a slide 50 on which there is a bar 51, the free end of which is adapted to resist forces for placement of the implant and to ensure that the approach is respected.","In the above description, the stabilised interbody fusion system 1 comprises an implant 4 made in the form of a generally parallelepiped shaped cage.","Obviously, the purpose of the invention could be used for an interbody implant in the form of a cylindrical or semi-cylindrical cage.","In the above description, the spacer bushing 31 is put into place during assembly of the stabilising plate 17 on the implant 4, while being carried by the assembly screw 22.Obviously, it will be possible for the spacer means 30 to be fixed onto the stabilising plate 17.Similarly, FIG.","7 illustrates another variant embodiment of the invention in which the spacer bushing 31 is fixed to the implant 4 before the operation.","This spacer bushing 31 comprises assembly means 55 on the implant, for example made of notches cooperating with complementary notches 56 formed on the anterior wall 6, and into which the threaded hole 23 opens up.","Therefore this spacer bushing 31 can then be installed on the implant 4 with the possibility of adjusting its degree of penetration, so that the distance between the implant 4 and the stabilising plate 17 can thus be adjusted.","Preferably, this spacer bushing 31 is split along its length so as to block its position during assembly of the assembly screw 22 that passes through this bushing 31, to be screwed into the threaded hole 23.The invention is not limited to the examples described and illustrated since many modifications can be made without departing from its scope."]],"string":"[\n [\n \"Stabilised interbody fusion system for vertebrae\",\n \"The invention concerns a stabilised interbody fusion system for vertebrae, of the type comprising an interbody implant (4) designed to be inserted in the intervertebral space defined between two neighbouring vertebrae to be mutually secured, so as to restore the height and the angle of the lordosis of the vertebral segment defined by the two neighbouring vertebrae and a stabilizing plate (17) provided, at each of its ends, with at least a passage hole (18) for an anchoring screw, the plate (17) and the implant (4) being provided with mutual assembly means, such that after assembly, the stabilizing plate (17) extends on each side of the implant to enable the stabilizing plate to be anchored on the neighbouring vertebrae through the screws, characterised in that it comprises spacing means (30), interposed between the stabilizing plate (17) and the implant (4), to enable the stabilizing plate to be positioned at a specific distance relative to the implant.\"\n ],\n [\n \"1-14.\",\n \"(canceled) 15.Stabilized interbody fusion system for vertebrae according to the invention, of the type comprising: an interbody implant that will be inserted in the intervertebral space defined between two neighboring vertebrae to be mutually secured, in order to restore the height and the angle of the lordosis of the vertebral segment defined by two neighboring vertebrae; and a stabilizing plate equipped with at least one passage hole for an anchoring screw at each of its ends, the plate and the implant being provided with mutual assembly means such that after assembly, the stabilizing plate extends on each side of the implant to enable the stabilizing plate to be anchored on the neighboring vertebrae through the screws, wherein it comprises spacing means, interposed between the stabilizing plate and the implant, to enable the stabilizing plate to be positioned at a specific distance relative to the implant.\",\n \"16.System according to claim 15, wherein the spacing means adjust the distance between the stabilizing plate and the implant.\",\n \"17.System according to claim 15, wherein the spacing means are chosen from among a range of spacing means each with a determined length, each of which is different, to adjust the distance between the plate and the implant at a value that is determined by the length of the chosen spacing means.\",\n \"18.System according to claim 15, wherein the assembly means are provided with support means for the plate allowing angular orientation of the plate relative to the implant.\",\n \"19.System according to claim 16, wherein the assembly means are provided with support means for the plate allowing angular orientation of the plate relative to the implant.\",\n \"20.System according to claim 17, wherein the assembly means are provided with support means for the plate allowing angular orientation of the plate relative to the implant.\",\n \"21.System according to claim 15, wherein the spacing means comprises at least one spacer bushing.\",\n \"22.System according to claim 16, wherein the spacing means comprises at least one spacer bushing.\",\n \"23.System according to claim 17, wherein the spacing means comprises at least one spacer bushing.\",\n \"24.System according to claim 18, wherein the spacing means comprises at least one spacer bushing.\",\n \"25.System according to claim 21, wherein the spacer bushing is fixed to the stabilizing plate.\",\n \"26.System according to claim 25, wherein the spacer bushing comprises assembly means on the implant, allowing adjustment of the distance between the stabilizing plate and the implant.\",\n \"27.System according to claim 25, wherein the spacer bushing is put into place on an assembly screw forming part of the assembly means and is designed to pass through the plate through a passage hole bordered by a stop shoulder arranged to hold the head of the assembly screw and to cooperate with at least one threaded hole arranged in the implant.\",\n \"28.System according to claim 27, wherein the threaded hole is formed on the anterior wall of the implant, this wall having a convex curvature in the transverse plane and extending by connecting walls with a convex curvature, to a posterior wall with a concave curvature to leave the vertebral canal free.\",\n \"29.System according to claim 28, wherein each connecting wall is arranged to contain a threaded hole surrounded, like the threaded hole formed in the anterior wall, by an assembly groove designed to cooperate with two assembly dog pins provided on the head of a gripping instrument of the implant, also comprising a threaded rod designed to cooperate with a threaded hole in the implant.\",\n \"30.System according to claim 28, wherein the posterior wall is fitted with a radio-opaque element opening up on the external face of the said wall, adapted so that the position of the implant within the intervertebral space along the sagittal plane can be detected.\",\n \"31.System according to claim 15, wherein the stabilizing plane is provided with anti-expulsion means for the anchoring screws, installed on the plate through moving guide means and adapted to occupy a first position in which the passage cross-section of the holes for reception of the anchoring screws is left free, and a second position, in which the holes are partially closed off, in order to form a stop for the heads of screws.\",\n \"32.A gripping instrument, comprising: a maneuvering tube, inside which a flexible threaded rod is fitted so as to cooperate with a threaded hole in the implant of claim 15, the tube further comprising an assembly head, fitted with two dog pins to cooperate with an assembly groove surrounding the threaded hole.\",\n \"33.The gripping instrument according to claim 32, wherein the maneuvering tube on the implant has a curvature and is provided with a slide on which there is a bar, the free end of which is adapted to resist forces for placement of the implant and to ensure that the approach is respected.\"\n ],\n [\n \"The present invention relates to interbody implants to be inserted in the intervertebral space defined between two neighbouring vertebrae after intervertebral disk excision, in order to restore the intervertebral space and to form bone fusion between the said neighbouring vertebrae.\",\n \"More precisely, the domain of the invention is lumbar type interbody implants to restore the height and the angle of the lordosis of the vertebral segment defined by two neighbouring lumbar vertebrae to be mutually secured.\",\n \"More precisely, the purpose of the invention is an interbody implant, called a secured implant, in other words provided with the means of anchoring the implant in its insertion site, to prevent it from becoming mobile.\",\n \"There are several stabilised interbody fusion systems for vertebrae known in the state of the art.\",\n \"For example, there is a known stabilised system comprising an interbody implant to be inserted in the intervertebral space defined between two neighbouring lumbar vertebrae.\",\n \"According to one preferred embodiment, this implant is in the form of a cage comprising two sagittal walls connected to each other through an anterior transverse wall and a posterior transverse wall.\",\n \"The walls delimit a volume between them opening up on each side of the transverse faces of the implant.\",\n \"The open volume of the cage will receive a bone filling product called a bone graft, that will come into contact with the vertebral plates to facilitate bone fusion between the two vertebrae.\",\n \"This type of stabilised system also comprises a stabilising plate extending from the anterior wall of the implant and on each side of the transverse faces.\",\n \"This stabilising plate is provided with passage holes at each of its ends for screws that will be anchored in the vertebrae to be mutually secured.\",\n \"A stabilising plate can be placed to reliably prevent migration of the implant.\",\n \"However, the applicant has identified a difficulty in keeping the implant in its ideal position within the intervertebral space to enable it to be anchored on the vertebrae to be mutually assembled through the screws.\",\n \"It must be borne in mind that, in different patients, firstly the anterior edges of the plates of the vertebra bodies have different profiles, and secondly implants are located at different distances from the anterior edges of the plates of the vertebra bodies, that causes displacement of the implant while the stabilising plate is anchored on the vertebrae.\",\n \"The applicant expressed the need to have a stabilised interbody fusion system designed firstly so that the implant can be placed in the intervertebral space in an appropriate position to restore a physiological spinal curvature and lordosis, and secondly to hold the implant in this ideal correction position during anchorage of the stabilising plate on the vertebrae, regardless of the shape of the anterior edges of the plates of the vertebra bodies and the position of the implant relative to these anterior edges of the plates.\",\n \"Therefore, the purpose of the invention is to satisfy this need by proposing a stabilised interbody fusion system capable of restoring the height and the angle of the lordosis of the vertebral segment by an implant, the function of which is secured by a stabilising plate fixed on the vertebrae.\",\n \"In order to achieve this objective, the stabilised interbody fusion system for vertebrae according to the invention is of the type comprising: an interbody implant that will be inserted in the intervertebral space defined between two neighbouring vertebrae to be mutually secured, in order to restore the height and the angle of the lordosis of the vertebral segment defined by two neighbouring vertebrae, and a stabilising plate equipped with at least one passage hole for an anchoring screw at each of its ends, the plate and the implant being provided with mutual assembly means such that after assembly, the stabilising plate extends on each side of the implant to enable the stabilising plate to be anchored on the neighbouring vertebrae through the screws, The stabilised system comprises spacing means interposed between the stabilising plate and the implant, to enable the stabilising plate to be positioned at a specific distance relative to the implant.\",\n \"Various other special features will become clear from the description given below with reference to the appended drawings that show example embodiments of the purpose of the invention as non-limitative examples.\",\n \"FIG.\",\n \"1 is an exploded perspective view showing the different constituents of the stabilised system according to the invention.\",\n \"FIG.\",\n \"2 is a sectional-elevation view of a stabilised system in the raised position.\",\n \"FIG.\",\n \"3 is an elevation view showing a stabilised system installed on two vertebrae.\",\n \"FIG.\",\n \"4 is a sectional-elevation view showing another variant embodiment of the stabilised system according to the invention.\",\n \"FIG.\",\n \"5 and 6 are perspective views showing characteristic details of a gripping instrument for an implant according to the invention.\",\n \"FIG.\",\n \"7 is a sectional-elevation view illustrating another variant embodiment of a stabilised system according to the invention.\",\n \"As is clear particularly in FIG.\",\n \"1 to 3, the invention relates to a stabilised interbody fusion system 1 for two neighbouring or nearby vertebrae 2, 3.The stabilised system 1 comprises an interbody implant 4 that will be inserted into the intervertebral space 5 delimited by the two vertebrae 2, 3.In one preferred embodiment of the invention as illustrated on the drawings, the implant 4 is made by a cage with a transverse wall 6 called the anterior wall, and a transverse wall 7 called the posterior wall, connected to each other by connecting walls 8, 9.The walls 6 to 9 of the cage are perpendicular to a transverse plane T that is perpendicular to a sagittal plane S passing through the anterior wall 6 and the posterior wall 7.In the transverse plane T, the anterior wall 6 has a convex curvature and extends on each side by the connecting walls 8, 9, which also have convex curvatures and are connected to each other by the posterior wall 7 with a concave curvature to leave the vertebral canal free.\",\n \"The cage 1 thus has a general kidney shape.\",\n \"As can be seen particularly in FIG.\",\n \"2, the connecting walls 8, 9 in the sagittal plane S converge towards each other towards the posterior wall 7, such that the cage 1 has a tapered section in the sagittal plane S defining a lordosis correction angle.\",\n \"The cage 1 comprises an interior volume 11 delimited by interior vertical faces of the walls 6 to 9 and will be filled by a bone filling product called a bone graft adapted to interbody fusion.\",\n \"This volume 11 opens up on a first transverse face 12 called the upper face in the example illustrated, and a second transverse face 13 called the lower face.\",\n \"These transverse faces 12, 13 are delimited by edges of the walls 6 to 9, preferably arranged to comprise notches 14 enabling the cage to be bonded to the supedjacent and subjacent vertebrae.\",\n \"The notches 14 extend parallel to each other and from a frontal plane F perpendicular to the sagittal plane S and the transverse plane T. Preferably, the internal volume 11 of the cage comprises one or several connecting walls 15 with other walls.\",\n \"In the example illustrated, the connecting wall 15 is in the general shape of a “Y” and extends between the connecting walls 8 and 9 and the anterior wall 6.The stabilised interbody fusion system 1 conform with the invention also comprises a stabilising plate 17 for the implant 4.This stabilising plate 17 is provided with at least one passage hole at each of its ends, and in the example illustrated with two passage holes 18 for the screws 19 that will be anchored on vertebrae 2 and 3 to be mutually secured.\",\n \"According to one preferred characteristic embodiment, the stabilising plate 17 is provided with anti-expulsion means 20 for the anchoring screws 19.These anti-expulsion means 20 are composed of moving flaps installed on the stabilising plate so that they can be guided in rotation in order to occupy a first position in which the passage cross-section of the holes 18 is left free, and a second position in which this passage cross-section is closed off in order to form a stop for the heads of screws 19.The stabilising plate 17 and the implant 4 are provided with mutual assembly means 21.The stabilising plate 17 is adapted so that after assembly with the implant 4, it extends from each side of the transverse faces 12, 13 to the other, to anchor the screws 19 on the vertebrae 2, 3.According to one characteristic embodiment, the assembly means 21 consist of an assembly screw 22 that will cooperate with at least one tapped hole 23 formed in the implant 4.For example, a tapped hole 23 is arranged to pass through the anterior wall from one side to the other along an axis perpendicular to the anterior wall 6 and centred on the sagittal plane S of symmetry of the implant 4.Preferably, the implant 4 comprises a series of threaded holes 23 to enable assembly of the stabilising plate 17 in different positions with respect to the implant 4.Thus, a threaded hole 23 is formed in a connecting wall, for example 9, along an axis perpendicular to the axis of the threaded hole 23 formed in the anterior wall 6.At the positions defined by these two threaded holes 23, the stabilising plate 17 is free to occupy two positions perpendicular to each other.\",\n \"Another threaded hole 23 may also be formed in the other connecting wall, 8, at an angle of 45° from the axis of the threaded hole 23 formed in the anterior wall 6.The assembly screw 22 passes through the stabilising plate 17 through a passage hole 24, bordered by a stop shoulder 25 formed by a dish 26 arranged to hold the head 221 of the assembly screw 22.According to one characteristic of the invention, the stabilised device 1 comprises spacing means 30 inserted between the stabilising plate 17 and the implant 4, to enable the stabilising plate 17 to be positioned at a specific distance relative to the implant 4.These spacing means 30 fix the stabilising plate 17 onto the vertebrae leaving the implant 4 in its position, taking account of the shape of the anterior edges B of the plates of vertebrae bodies 2, 3 or the setback position of the implant 4 relative to these anterior edges of the vertebrae.\",\n \"In the example illustrated in FIG.\",\n \"1 to 3, the spacing means 30 are composed of a spacer bushing 31 provided with a thread 32 cooperating with the thread of the assembly screw 22.The spacer bushing 31 is thus made in the form of a nut and its outside shape is prismatic and is adapted to cooperate with a part 33 for reception of the implant 4 in which the threaded hole 23 opens up.\",\n \"Cooperation of the spacer bushing 31 with the reception part 33 blocks the bushing 31 in rotation when the assembly screw 22 is being screwed.\",\n \"Preferably, the reception part 33 is made by an entry cone cooperating with a complementary part 34 presented by the bushing 31.Cooperation of the bushing 31 with the entry cone 33 is a means of adjusting the distance between the stabilising plate 17 and the implant 4.The stabilised system 1 according to the invention is used directly as a function of the description given above.\",\n \"Before the implant 4 is inserted, the disc is excised by preparation of the vertebral plates.\",\n \"The implant 4 is implanted in the intervertebral space 5 between two adjacent vertebrae 2 and 3 to be mutually secured.\",\n \"The implant 4 is designed to restore the height and the angle of the lordosis of the vertebral segment defined by the two neighbouring vertebrae 2, 3.The stabilising plate 17, fitted with its assembly screw 22 provided with its spacer bushing 31, will be installed on the implant 4.The screw 22 is then screwed into a threaded hole 23 of the implant.\",\n \"The screw 22 is then screwed in until it is fixed in place by the spacer bushing 31 being stopped between the plate 17 and the implant 4.The screws 19 are then anchored in the vertebrae 2, 3.Obviously, the length of the spacer bushing 31 is adapted to allow the implant 4 to remain in its implantation site, while allowing the stabilising plate 17 to come into contact with the vertebrae.\",\n \"The stabilising plate 17 and the implant 4 are separated by a fixed measurement determined by the spacer bushing 31.Preferably, a range of spacer bushings 31 is available.\",\n \"Each of these spacer bushings 31 has a determined length, each of which is different, to adjust the distance between the implant 4 and the stabilising plate 17.Note that the implant 4 is preferably provided with at least one radio-opaque element 35 (FIGS.\",\n \"1 and 2) adapted so that the position of the implant 4 within the intervertebral space 5 along the sagittal plane S can be detected on an X-ray.\",\n \"In the example illustrated, the radio-opaque element 35 is installed in the posterior wall 7 extending in the transverse plane and opening up on the external face and the internal face of the posterior wall 7.According to one example embodiment illustrated in FIG.\",\n \"4, the assembly means 21 are provided with support means 36 for the stabilising plate 17 allowing angular orientation of the stabilising plate relative to the implant 4.For example, these support means 36 may be made by a ball joint type articulation formed between the head 221 of the assembly screw 22 and the dish 26 in the plate 17.In this variant embodiment illustrated, the screw 22 is assembled to the stabilising plate 17 using a ring 38 fixed on the plate 17 to trap the head 221 of the screw 22, while allowing the screw 22 free to rotate about its own axis.\",\n \"According to one preferred variant embodiment, each threaded hole 23 of the implant is surrounded by an assembly groove 40, inclined relative to the sagittal plane and designed to cooperate with a gripping or manipulation instrument 44 for use on the implant 4.As can be seen in FIGS.\",\n \"5 and 6, the manipulation instrument 44 comprises an assembly head 45 prolonged by a manoeuvring tube 46 inside which a flexible threaded rod 47 passing through the head 45 is installed, so as to cooperate with a threaded hole 23 in the implant.\",\n \"The assembly head 45 is provided with two dog pins 49 that will be inserted in the groove 40 on each side of the threaded hole 23.The instrument 44 thus enables complete gripping and blocking of the implant 4 with respect to the assembly head 45.According to one preferred variant embodiment, the manoeuvring tube 46 has a curvature and is provided with a slide 50 on which there is a bar 51, the free end of which is adapted to resist forces for placement of the implant and to ensure that the approach is respected.\",\n \"In the above description, the stabilised interbody fusion system 1 comprises an implant 4 made in the form of a generally parallelepiped shaped cage.\",\n \"Obviously, the purpose of the invention could be used for an interbody implant in the form of a cylindrical or semi-cylindrical cage.\",\n \"In the above description, the spacer bushing 31 is put into place during assembly of the stabilising plate 17 on the implant 4, while being carried by the assembly screw 22.Obviously, it will be possible for the spacer means 30 to be fixed onto the stabilising plate 17.Similarly, FIG.\",\n \"7 illustrates another variant embodiment of the invention in which the spacer bushing 31 is fixed to the implant 4 before the operation.\",\n \"This spacer bushing 31 comprises assembly means 55 on the implant, for example made of notches cooperating with complementary notches 56 formed on the anterior wall 6, and into which the threaded hole 23 opens up.\",\n \"Therefore this spacer bushing 31 can then be installed on the implant 4 with the possibility of adjusting its degree of penetration, so that the distance between the implant 4 and the stabilising plate 17 can thus be adjusted.\",\n \"Preferably, this spacer bushing 31 is split along its length so as to block its position during assembly of the assembly screw 22 that passes through this bushing 31, to be screwed into the threaded hole 23.The invention is not limited to the examples described and illustrated since many modifications can be made without departing from its scope.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473773"}}},{"rowIdx":753,"cells":{"text":{"kind":"list like","value":[["Actuator assembly","An actuator assembly comprises a first member (6a,6b), a second member (4a,4b) and at least one connecting arm (10a,10b) of fixed length connecting the first and second members (6a,6b;4a,4b).","The assembly also comprises an actuator (12,14) which is operably engaged with the first member (6a,6b) so as to be capable of applying a rotating force to the first member (6a,6b).","In use, rotational movement of the first member (6a,6b) causes relative linear movement of the first and second members (6a,6b;4a,4b) along an axis.","The invention also relates to a loudspeaker driver unit and a sensor incorporating the actuator assembly.","In the sensor embodiments, the actuator is an electrical transducer."],["1.A method implemented by a push-to-talk wireless mobile terminal for communicating voice information comprising the steps of: determining if a request to send a delayed delivery voice message has been made; if said determining step determines that a request to send a delayed delivery voice message has been made, transmitting an indicator to a communication application server representing an instruction that packets received from the mobile terminal are to be stored for later delivery to a destination Pal; encoding audio input from a user by the mobile terminal into the packets following the determining step; transmitting the packets to the communication application server for later delivery to the destination Pal.","2.The method of claim 1 wherein the step of determining if a request to send a delayed delivery voice message has been made comprises sensing that the destination Pal selected by the user to receive the audio input is not available prior to the user initiating the encoding step.","3.The method of claim 2 wherein the step of sensing that the Pal selected by the user as the destination party to receive the audio input is not available comprises the step of determining a current status of the selected Pal stored in the mobile terminal, where the status represents that the selected Pal is not available to receive communications.","4.The method of claim 1 wherein the step of determining if a request to send a delayed delivery voice message has been made comprises determining that an input signal is entered by the user where the input signal represents a request to send said packets as a delayed delivery voice message regardless of whether or not the destination Pal selected to receive the packets is currently available to receive communications.","5.The method of claim 1 wherein the step of transmitting an indicator to a communication application server representing an instruction that the packets are to be stored for later delivery to a destination Pal causes the audio carried by the packets to be stored in the communication application server.","6.The method of claim 5 wherein the instruction further conveys to the communication application server that the packets are not to be attempted to be delivered in real-time to the destination Pal.","7.The method of claim 1 wherein the step of determining if a request to send a delayed delivery voice message has been made comprises the steps of generating a first request for a real-time voice communication to the destination Pal in response to the push-to-talk button being pressed, providing a first alert to the user indicating that a real-time voice communication to the destination Pal is not available, and sensing a second request to initiate communications to the destination Pal by the push-to-talk button being depressed again following the alert having been provided to the user.","8.The method of claim 7 wherein the step of sensing the second request includes sensing the push-to-talk button being depressed again within a predetermined time interval of the first request.","9.The method of claim 7 further comprising the step of providing a second alert to the user in response to the second request wherein the second alert comprises a predetermined talk-beep associated with the request to send a delayed delivery voice message, the predetermined talk-beep comprising an audible alert that is different from an audible alert associated with the initiation of a real-time voice communication.","10.A push-to-talk wireless mobile terminal for communicating voice information comprising: means for determining if a request to send a delayed delivery voice message has been made; means for transmitting an indicator to a communication application server representing an instruction that packets received from the mobile terminal are to be stored for later delivery to a destination Pal if said determining means determines that a request to send a delayed delivery voice message has been made; means for encoding audio input from a user by the mobile terminal into the packets following the determining step; means for transmitting the packets to the communication application server for later delivery to the destination Pal.","11.The mobile terminal of claim 10 wherein the means for determining comprises means for sensing that the destination Pal selected by the user to receive the audio input is not available to receive real-time communications.","12.The mobile terminal of claim 11 wherein the means for sensing comprises means for determining a current status of the selected Pal stored in the mobile terminal, where the status represents that the selected Pal is not available to receive communications.","13.The mobile terminal of claim 10 wherein the means for determining if a request to send a delayed delivery voice message has been made comprises means for determining that an input signal is entered by the user where the input signal represents a request to send said packets as a delayed delivery voice message regardless of whether or not the destination Pal selected to receive the packets is currently available to receive communications.","14.The mobile terminal of claim 10 wherein the means for transmitting an indicator to a communication application server representing an instruction that the packets are to be stored for later delivery to a destination Pal causes the audio carried by the packets to be stored in the communication application server.","15.The mobile terminal of claim 14 wherein the instruction transmitted by the means for transmitting further conveys to the communication application server that the packets are not to be attempted to be delivered in real-time to the destination Pal.","16.The mobile terminal of claim 10 wherein the means for determining if a request to send a delayed delivery voice message has been made comprises means for generating a first request for a real-time voice communication to the destination Pal in response to the push-to-talk button being pressed, means for providing a first alert to the user indicating that a real-time voice communication to the destination Pal is not available, and means for sensing a second request to initiate communications to the destination Pal by the push-to-talk button being depressed again following the alert having been provided to the user.","17.The mobile terminal of claim 16 wherein the means for sensing the second request includes means for sensing the push-to-talk button being depressed again within a predetermined time interval of the first request.","18.The mobile terminal of claim 16 further comprising means for providing a second alert to the user in response to the second request, the second alert comprises a predetermined talk-beep associated with the request to send a delayed delivery voice message, the predetermined talk-beep comprising an audible alert that is different from an audible alert associated with the initiation of a real-time voice communication.","19.A method implemented by a push-to-talk wireless mobile terminal for communicating voice information comprising the steps of: displaying a list of Pals of the first user including visual indicia representing that a previously transmitted voice message by the first user to a first Pal is stored and awaits delivery to the first Pal; determining if a request has been made by the first user to access the stored voice message; if said request is determined to have been made, discerning the type of access desired by the first user; transmitting a command to a communication application server at which the voice messages stored where the command conveys instructions to the communication application server to implement action based on the type of access desired by the first user.","20.The method of claim 19 wherein the step of discerning comprises discerning that the stored voice message is to be played back to the first user and wherein the transmitted command conveys instructions to the communication application server to implement transmission of the stored voice message to the first user, the method further comprising the step of receiving at the mobile terminal of the first user the stored voice message previously transmitted by the first user to the first Pal and audibly playing the stored voice message.","21.The method of claim 19 wherein the step of discerning comprises discerning that the stored voice message is to be deleted and wherein the transmitted command conveys instructions to the communication application server to delete the stored voice message previously transmitted by the first user to the first Pal, the method further comprising the step of receiving at the mobile terminal of the first user a signal technology that the stored voice message has been deleted.","22.A push-to-talk wireless mobile terminal for communicating voice information comprising: means for displaying a list of Pals of the first user including visual indicia representing that a previously transmitted voice message by the first user to a first Pal is stored and awaits delivery to the first Pal; means for determining if a request has been made by the first user to access the stored voice message; means for discerning the type of access desired by the first user if said request is determined to have been made; means for transmitting a command to a communication application server at which the voice messages are stored where the command conveys instructions to the communication application server to implement action based on the type of access desired by the first user.","23.The mobile terminal of claim 22 wherein the means for discerning discerns that the stored voice message is to be played back to the first user and wherein the means for transmitting transmits the command that conveys instructions to the communication application server to implement transmission of the stored voice message to the first user, the mobile terminal further comprising means for receiving, at the mobile terminal of the first user, the stored voice message previously transmitted by the first user to the first Pal and audibly playing the stored voice message.","24.The mobile terminal of claim 22 wherein the means for discerning discerns that the stored voice message is to be deleted and wherein the means for transmitting transmits the command that conveys instructions to the communication application server to delete the stored voice message previously transmitted by the first user to the first Pal, the mobile terminal further comprising means for receiving at the mobile terminal of the first user a signal representing that the stored voice message has been deleted.","25.A method implemented by a push-to-talk wireless mobile terminal for communicating voice information comprising the steps of: displaying a list of Pals including visual indicia of whether a voice message is waiting for delivery from a Pal; determining if a request to receive a waiting voice message has been initiated by a user of the mobile terminal; if said request is determined to have been made, transmitting at least one packet to a communication application server requesting that the waiting voice message associated with a selected Pal be delivered to the user's mobile terminal; receiving packets at the user's mobile terminal from the communication application server containing the waiting voice message and playing the message to the user.","26.The method of claim 25 wherein the steps of determining if the request has been made comprises sensing that a Pal is selected by the user where the Pal has corresponding visual indicia indicating that a voice message from the Pal is waiting delivery to the user, and sensing an input initiated by the user representing a request to receive delivery of the voice message corresponding to the selected Pal.","27.The method of claim 26 further comprising the steps of receiving a status update following receiving the packets where the status update indicates that there is no longer the voice message from the Pal waiting delivery to the user and updating the visual indicia corresponding to the Pal whose voice message was received to show another visual indicia representing that the voice message is no longer waiting delivery to the user.","28.A push-to-talk wireless mobile terminal for communicating voice information comprising: means for displaying a list of Pals including visual indicia of whether a voice message is waiting for delivery from a Pal; means for determining if a request to receive a waiting voice message has been initiated by a user of the mobile terminal; means for transmitting at least one packet to a communication application server requesting that the waiting voice message associated with a selected Pal be delivered to the user's mobile terminal if said request is determined to have been made; means for receiving packets at the user's mobile terminal from the communication application server containing the waiting voice message and playing the message to the user.","29.The mobile terminal of claim 28 wherein the means for determining if the request has been made comprises means for sensing that a Pal is selected by the user where the Pal has corresponding visual indicia indicating that a voice message from the Pal is waiting delivery to the user, and means for sensing an input initiated by the user representing a request to receive delivery of the voice message corresponding to the selected Pal.","30.The mobile terminal of claim 29 further comprising means for receiving a status update following receiving the packets where the status update indicates that there is no longer the voice message from the Pal waiting delivery to the user and means for updating the visual indicia corresponding to the Pal whose voice message was received to show another visual indicia representing that the voice message is no longer waiting delivery to the user.","31.A method implemented by a communication application server in a packet communication network for processing communications comprising the steps of: receiving a first packet from a mobile terminal of a user; determining if the first packet contains an indicator representing an instruction to process any following voice packets as a delayed delivery voice message; receiving a set of voice packets from a mobile terminal of a user; if said determining step determines that said indicator is present, storing at least payloads of the voice packets of said set in memory and labeling the stored payloads as addressed to a destination Pal identified by said set of packets.","32.The method of claim 31 further comprising the step of attempting to deliver voice information contained in said payloads only upon receiving a command from a mobile terminal of the destination Pal where the command corresponds to input initiated by the Pal to retrieve the stored voice information.","32.The method of claim 31 further comprising the steps of generating a status update following the storing step and transmitting the status update to at least the mobile terminals of the destination Pal and the user.","33.The method of claim 32 wherein the step of transmitting the status update comprises transmitting a status update to the destination Pal indicating that the voice information from the user is awaiting deliver to the destination Pal and transmitting a status update to the user indicating that the voice information from the user to the destination Pal is still awaiting delivery to the destination Pal.","34.The method of claim 32 further comprising the step of receiving said command from the mobile terminal of the destination Pal, retrieving the at least voice information payloads, encoding the at least voice information payloads into further packets addressed to the destination Pal, and transmitting the further packets to the destination Pal.","35.The method of claim 34 further comprising the steps of generating a status update following the transmitting of the further packets and transmitting the status update to at least the mobile terminals of the destination Pal and the user, where the status update represents that the voice information has been delivered to the destination Pal.","36.The method of claim 35 wherein the step of transmitting the status update comprises transmitting a status update to the destination Pal indicating that there is no longer the voice information from the user awaiting deliver to the destination Pal and transmitting a status update to the user indicating that there is no longer the voice information from the user awaiting delivery to the destination Pal.","37.A communication application server in a packet communication network for processing communications comprising: means for receiving a first packet from a mobile terminal of a user; means for determining if the first packet contains an indicator representing an instruction to process any following voice packets as a delayed delivery voice message; means for receiving a set of voice packets from a mobile terminal of a user; means for storing at least payloads of the voice packets of said set in memory and labeling the stored payloads as addressed to a destination Pal identified by said set of packets if said determining step determines that said indicator is present.","38.The communication application server of claim 37 further comprising means for attempting to deliver voice information contained in said payloads only upon receiving a command from a mobile terminal of the destination Pal where the command corresponds to input initiated by the Pal to retrieve the stored voice information.","39.The communication application server of claim 37 further comprising means for generating a status update following the storage of the at least payloads and means for transmitting the status update to at least the mobile terminals of the destination Pal and the user.","40.The communication application server of claim 39 wherein the means for transmitting the status update comprises means for transmitting a status update to the destination Pal indicating that the voice information from the user is awaiting delivery to the destination Pal and means for transmitting a status update to the user indicating that the voice information from the user to the destination Pal is still awaiting delivery to the destination Pal.","41.The communication application server of claim 39 further comprising means for receiving said command from the mobile terminal of the destination Pal, means for retrieving the at least voice information payloads, means for encoding the at least voice information payloads into further packets addressed to the destination Pal, and means for transmitting the further packets to the destination Pal.","42.The communication application server of claim 41 further comprising means for generating a status update following the transmitting of the further packets and means for transmitting the status update to at least the mobile terminals of the destination Pal and the user, where the status update represents that the voice information has been delivered to the destination Pal.","43.The communication application server of claim 42 wherein the means for transmitting the status update comprises means for transmitting a status update to the destination Pal indicating that there is no longer the voice information from the user awaiting deliver to the destination Pal and means for transmitting a status update to the user indicating that there is no longer the voice information from the user awaiting delivery to the destination Pal."],["The present invention relates, in a first aspect, to an actuator assembly in which a rotating force applied to a first member is converted into linear movement.","The present invention also relates to loudspeaker driver units comprising such a device.","There are many devices (such as loudspeaker driver units and focussing assemblies for cameras and microscopes etc and CD and DVD players) into which linear actuator assemblies are incorporated.","In such devices one or more of the following factors may be important: mass, power consumption, compactness and linearity of response.","For example, in magnet-based loudspeaker driver units, a moving coil oscillates in a magnetic field generated by a permanent magnet.","The magnet may make up as much as 90% of the mass of the driver unit, but serves no other purpose than to generate the magnetic field.","Crystal loudspeaker driver units are also known.","These incorporate a piezoelectric crystal which undergoes a change in thickness when subjected to a potential difference.","When an alternating voltage is applied, the crystal undergoes bending oscillations which are transmitted to a speaker diaphragm.","Crystal-based driver units are particularly suited to the generation of high frequencies, and a conventional loudspeaker may comprise an electrodynamic- and crystal-based driver unit in order to achieve a full range (20 Hz-20 kHz) frequency response.","Geophone sensors (see for example U.S. Pat.","No.","4,152,692), used to detect seismic vibrations, basically comprise a coil suspended between a pair of springs and positioned around a strong magnet.","Vertical linear travel of the coil due to seismic vibrations produces strong electrical signals in the coil.","Such magnet-coil based devices are generally complex and relatively heavy and large.","An object of a first aspect of the present invention is to provide a novel linear actuator assembly which obviates or mitigates one or more disadvantages of known actuator assemblies.","A further object of the present invention is to provide improved devices, such as a loudspeaker driver unit, incorporating such an assembly.","An object of a second aspect of the present invention is to provide an improved vibration sensor.","According to the present invention, there is provided an actuator assembly comprising:— (i) a first member, (ii) a second member, (iii) at least one connecting arm of fixed length connecting said first and second members, and (iv) an actuator operably engaged with said first member so as to be capable of applying a rotating force to the first member, wherein in use, rotational movement of the first member causes relative linear movement of the first and second members along an axis.","Preferably, the first member and actuator are arranged so that the force applied by the latter causes the former to rotate in a plane substantially perpendicular to said axis.","The actuator is preferably a piezoelectric or electrostrictive transducer.","Preferably, the actuator is in the form of a spiral having at least one half turn.","It will be understood that the greater the number of turns for a given actuator, the greater the maximum angular actuation will be.","Examples of suitable piezoelectric materials include ceramic materials such as lead-zirconate-titanate (PZT) based systems or non-ceramic systems (eg.","polymer based systems such as polyvinylidene fluoride).","Particularly preferred compositions are those classified by the US Department of Defence under DOD STD-1376A type VI.","An example of which is PZT-5H (sold by Morgan Electroceramics).","Preferably, the piezoelectric material has a lateral piezoelectric strain (d31) coefficient greater than 200 pC/N.","More preferably the d3coefficient is no more than 350 pC/N.","Preferably the elastic stiffness of the piezoelectric material is at least 65 GPa.","When a piezoelectric material is used it preferably has a bimorph or multimorph structure, although unimorph structures may be used.","If an electrostrictive material is used, it is preferably a ceramic material, and more preferably based on the lead magnesium niobate-lead titanate (PMN-PT) system.","Preferably, the first and second members are annular with differing diameters (“inner” and “outer” annular members) interconnected by at least two (but preferably three) connecting arms.","Preferably, said arms are arranged symmetrically between said annular members.","More preferably, said arms are arcuate.","The first member may be the inner or outer member.","Preferably, the first and second members and said at least one connecting arm are of unitary construction.","Such a construction in which inner and outer annular rings are interconnected by arcuate connecting arms will hereinafter be referred to as a “plate spring” or a “spiral arm spring”.","Such springs are per se known and have been used in geophone sensor units.","It will be understood that when the inner and outer rings of a plate spring are moved apart along an axis perpendicular to the plane of the spring, there must be relative rotation of the inner and outer rings because the connecting arms are of fixed length.","Consequently, if one ring is caused to rotate and the other is prevented from rotating, relative linear movement of the rings will be induced perpendicular to the plane of the spring.","It will also be understood from the foregoing that one of the first and second members may be mounted so as to prevent movement along the axis, in which case actuation will result in movement of the other member along the axis.","In a first series of embodiments, the first member is mounted so that linear movement along the axis is prevented, whereas the second member is mounted for linear movement along the axis.","In a preferred arrangement, the actuator (preferably in spiral form) is positioned inside the first (inner) annular member and secured thereto.","Securement may be achieved by, for example, soldering, fusing, or bonding with adhesive.","Suitable formations (eg.","tabs or flanges) onto which to secure the actuator may be provided on the first member.","Alternatively, the actuator (preferably in spiral form) is positioned outside the first (outer) annular member and secured thereto.","Such an arrangement permits the mounting of, for example, a lens inside the second (inner) annular member.","In a highly preferred embodiment, the actuator assembly comprises first and second plate springs whose outer annular rings are secured together (directly or indirectly, eg.","by placing a stiffening ring or cylindrical collar therebetween), and first and second spiral actuators arranged to actuate the respective inner rings of the first and second plate springs wherein the actuators are oppositely orientated so that, in use, actuation of the first spiral actuator rotates the inner ring of the first plate spring in one direction about the axis, whereas simultaneous actuation of the second actuator rotates the inner ring of the second plate spring in the opposite direction about the axis by an equal amount, whereby to move the outer rings along the axis.","It will be readily understood that the outer rings can be moved in either direction along the axis depending on the polarity of the applied voltage.","In a very highly preferred embodiment, the outer rings are secured together and the inner rings are equidistantly spaced either side of the outer rings.","In a second series of embodiments, the second member is mounted so that linear movement along the axis is prevented, whereas the first member and actuator are mounted for linear movement along the axis.","The present invention also resides in a loudspeaker driver unit and loudspeaker comprising an actuator assembly in accordance with the present invention and an air piston driven by the actuator assembly to generate an acoustic wave.","The air piston may be in the form of a hemisphere or a conical diaphragm.","Alternatively, the loudspeaker driver unit may include a diaphragm which is oscillated by the actuator assembly to generate an acoustic wave.","It will be understood that if the actuator is in fact an electrical transducer, rotation of the transducer will result in the generation of an electrical signal in the transducer.","Thus, according to a second aspect of the present invention, there is provided a sensor comprising:— (i) a first member, (ii) a second member, (iii) at least one connecting arm of fixed length connecting said first and second members and (iv) a transducer operably engaged with said first member, wherein in use, relative linear movement of the first and second members along an axis causes rotational movement of the first member which is transmitted to said transducer whereby to generate an electrical signal in said transducer.","Said sensor corresponds closely to said actuator assembly, the primary difference being that in the former, relative linear movement is converted into rotational movement and subsequently into an electrical signal, whereas in the latter, actuated rotational movement is converted into relative linear movement.","It will therefore be understood that the preferred features of the assembly of the first aspect are also preferred features of the sensor of the second aspect.","The sensor may be a vibration sensor, e.g.","for detecting seismic vibrations.","Embodiments of the present invention will now be described by way of example only, with reference to the accompanying drawings in which:— FIG.","1 is plan view of a plate spring suitable for use in the actuator assembly of the first aspect of the invention, or the sensor of the second aspect of the invention, FIG.","2 shows the triangle formed between an arm of length L lying on a diameter D when the spring of FIG.","1 is offset by a displacement 6, FIG.","3 is a graph of linear travel against relative rotation derived for the spring of FIG.","1, FIGS.","4 and 5 show an embodiment of an actuator assembly in accordance with the first aspect of the present invention, FIG.","6 is a graph of linear travel against relative rotation derived for the embodiment of FIGS.","4 and 5.FIGS.","7a to 7c are schematic representations of a loudspeaker driver unit incorporating an actuator assembly in accordance with the first aspect of the present invention in an extreme inner (FIG.","7a), intermediate (FIG.","7b) and extreme outer (FIG.","7c) position, FIGS.","8a to 8c correspond to FIGS.","7a to 7c for a stiffened loudspeaker driver unit incorporating an actuator assembly in accordance with the first aspect of the present invention, FIGS.","9a to 9c correspond to FIGS.","7a to 7c for a partially stiffened loudspeaker driver unit incorporating an actuator assembly in accordance with the first aspect of the present invention, and FIG.","10 is a schematic view of part of a vibration sensor in accordance with the second aspect of the invention.","Referring to FIG.","1, a plate spring 2 comprises a first (outer) annular ring 4 and a second (inner) annular ring 6, the first and second rings 4,6 being concentric (i.e.","coplanar) in the rest position of the spring 2.A pair of tabs 8 angularly spaced by 180° extends radially inwardly from the inner ring 6.The first and second rings are connected by three part-annular connecting arms 10.Each arm 10 subtends an angle of 0 and lies on a circle of diameter D. The plate spring 2 is of unitary construction and fabricated from beryllium copper alloy.","In use, the tabs 8 are bent out of the plane of the spring 2 and serve as mounting points for a spiral actuator (described below).","It will be understood that as the two rings 4,6 are moved relative to one another along an axis perpendicular to the plane of the spring 2, there must be relative rotation of the rings 4,6 to maintain the rings 4,6 in a parallel orientation.","This is because the connecting arms 10 are of fixed length.","An approximate relationship between the linear movement and the relative rotation can be derived as follows:— The length of each arm 10 is given by L = D θ 2 If the inner and outer rings 4,6 are displaced perpendicularly to the plane of the spring 2 relative to each other by a distance δ, the angle subtended by each arm 10 (as viewed in FIG.","1) must change.","This can be calculated approximately by assuming that the arms 10 lie on the same diameter (D) and form perfect helical lines connecting the inner and outer rings 4,6.FIG.","2 shows the triangle formed by the arc length L and the displacement 6.The projection of the arc on to the initial plane of the spring 2 is then given by L = D 2 ( θ - Δ θ ) where Δθ corresponds to the relative rotation of the inner and outer rings 4,6 of the spring 2 necessary for them to remain parallel, with L being constant.","The displacement δ can then be given as θ = D 2 Δ θ 2 + 2 θ Δ θ This relationship is depicted in FIG.","3 from which it will be noted that at relatively large linear displacements, the relationship between linear travel and rotation is approximately linear.","The present invention takes advantage of this effect which allows linear movement to be produced directly from a rotational actuation.","Referring to FIGS.","4 and 5, an embodiment of the actuator assembly comprises first and second identical plate springs 2a,2b, first and second identical piezoelectric ceramic actuators 12, 14 and a mounting post 16.The plate springs 2a,2b are similar to that described with reference to FIG.","1 (and the same reference numerals are used to denote corresponding structures, suffixed by “a” and “b” to denote the first and second springs respectively), but there are only two connecting arms 10.Each actuator 12, 14 is formed from a tape of a lead-zirconate-titanate (PZT) composition having a bimorph structure which is wound into a spiral having 4 turns in the present embodiment.","Such piezoelectric ceramic materials are particularly suited to the present invention because they can exhibit a lateral piezoelectric strain (d31) coefficient as high as 350 pC/N, while possessing a flexural elastic modulus of over 60 Gpa.","If only small actuation movements are required, these properties allow high forces to be generated from a small amount of material.","This is useful in certain applications, such as in loudspeaker driver units as will be described below.","The outer diameter of the actuator spirals 12, 14 corresponds to the inner diameter of the inner rings 6a,6b of the plate springs 2a,2b.","A Bimorph piezoelectric structure is formed from two layers of piezoelectric material, separated by a conductive central electrode.","Electrodes are placed on the outer surfaces of the ceramic layers, and the layers are poled and actuated using these three electrodes such that the overall effect of the actuation is to expand one ceramic layer while causing the other to contract, through the effect of the d31 coefficient, thus producing a uniform bending strain in the element.","Various methods for creating ceramic compositions suitable for use in the present invention are known, see for example EP01 83453, EP0288208 and N. Alford et al, Nature; v.330; pp 51-53.To form the required spiral actuator structure, it is most beneficial to first create the required Bimorph structure in a planar form.","This may be done through the routes of printing and lamination.","A green (unfired) ceramic tape is formed from PZT powder mixed with a polyvinyl butyral (PVB) binder and cyclohexanone solvent.","In the present embodiment the formulation is 100 parts by weight of PZT to 6 parts PVB, to 7 parts cyclohexanone and 0.1 parts stearic acid, the stearic acid serving as a surfactant.","The green tape is then printed with the internal electrode, which may be of platinum, silver or an alloy of silver and palladium, formed into a printable ink.","In the present embodiment platinum is used (grade C51121D1 supplied by Gwent Electronic Materials, Pontypool).","The printed tape is then laminated with another ceramic tape of the same type and thickness (in the present embodiment PZT-5H, each tape 0.35 mm thick in the green state).","The lamination step may involve pressure and/or heat to achieve a strong bond across the electrode print.","The outer electrodes are then printed in the same fashion as the internal electrode, and allowed to dry.","After the printing stage, the overall tape structure must be sufficiently flexible and plastic to be deformed into the required spiral actuator structure.","This shaping may be achieved by using a tape formation route which includes a thermoplastic binder, in which case heat and pressure may be used to deform the tape into the required spiral.","With a solvent and binder system as in the present embodiment, the presence of the solvent allows the material to remain plastically deformable prior to removal of the solvent through evaporation.","An interleaving tape may be used, in order to maintain separation of the spiral turns during shaping.","This material may be in the form of carbon, formed in the same manner as the ceramic tape.","In the present embodiment 35 parts by weight of carbon black, 11 parts PVB and 12 parts cyclohexanone are used.","After the plastic processing step, and after drying, if applicable, the carbon tape is removed along with the binder in the ceramic tape through slow heating up to 600° C. in air.","Extra interleaving layers may be used between the PZT and the carbon layers to prevent the tapes adhering to each other while the solvent is still present.","Suitable materials include polythene, preferably less than 50:m thick.","This may be removed from the spirals after drying.","The spiral form is then sintered in an enclosed crucible with sufficient excess PbO-containing material, such as lead zirconate, to prevent PbO loss from the piezoelectric material.","After sintering, the thickness of the tapes is reduced to about 0.3 mm giving a total actuator thickness of 0.6 mm.","Soldered electrical connections are then made to the three separate electrode layers, with a wire connected to each.","The outer two layers of the tape are connected to a high voltage supply, and the device is placed in a heated oil bath at 120-130° C. A voltage equivalent to 2.5 kV/mm across the whole tape thickness is applied while the device is in the bath for 10 minutes.","This process polarises the piezoelectric tape.","After polarisation, the third electrode, connected to the central electrode layer, can be used to apply a field which is in opposite directions on each half of the tape.","The outer two electrodes can therefore be connected together, and used as the ground electrode, while the central electrode can be used for the driving signal.","For driving, the opposing electric fields generate bending in the tape.","Each of the actuator spirals 12, 14 is securely mounted onto the mounting post 16, each actuator 12, 14 being oppositely orientated relative to the other with the actuators 12, 14 spaced a short distance apart.","The outer rings 4a,4b of the springs 2a,2b are securely soldered to opposite sides of a stiffening ring (not shown) which prevents warping of the outer rings 4a,4b.","It should be noted that the springs 2a,2b are in the same orientation.","Each spring 2a,2b is tensioned by moving the inner ring 6a,6b out of the initial plane of the spring 2a,2b.","The inner ring 6a of the first spring 2a is moved towards the end of the first actuator 12 remote from the second actuator 14 where it is securely fixed via the tabs (8, FIG.","1) to the outer curved surface of the first actuator 12 by soldered joints.","The tabs lie 180° apart so that the axial forces on the outer rings 4a,4b do not produce unbalanced forces on the actuator spirals 12, 14.The inner ring 6b of the second spring 2b is moved in the opposite direction (i.e.","towards the end of the second actuator 14 remote from the first actuator 12) where it is secured to the curved surface of the second actuator 14 via the tabs (8, FIG.","1) to the outer curved surface of the second actuator 14 by soldered joints.","As can be seen in FIG.","4, the soldered outer rings 4a,4b of the springs 2a,2b are equidistant from the respective inner rings 6a,6b.","In use, when an electrical signal is applied to both actuator spirals 12,14 in parallel via the respective inner and outer electrodes connected to a power source (not shown), the actuators 12, 14 “rotate” in opposite directions.","The rotation of the first actuator 12 is transmitted to the inner ring 6a of the first spring 2a and the opposite rotation of the second actuator 14 is transmitted to the inner ring 6b of the second spring 2b.","Since the inner rings 6a,6b of the springs 2a,2b are prevented from translational movement by means of their securement to the respective actuator 12, 14, the outer rings 4a,4b move along an axis perpendicular to the planes of the springs 2a,2b.","This is the only way in which the rotational force can be transmitted, since the first and second outer rings 4a,4b, although capable of rotation, are subjected to equal but opposite rotational forces from the respective actuators 12, 14.No rotation results because of their securement to the stiffening ring.","Thus, it will be understood that the two actuators 12, 14 work in unison.","Depending on the overall design of the device, the travel of the outer rings 4a,4b may be anything from about 100 μm up to several centimetres.","If the polarity of the applied voltage is reversed, the outer rings 4a,4b will move along the axis in the opposite direction.","The outer rings 4a,4b can be moved between the respective planes of the inner rings 6a,6b of the first and second springs 2a,2b, the exact position of the outer rings 4a,4b being dependent on the polarity and magnitude of the applied voltage to the actuators 12, 14.The actuator assembly is self centring, i.e.","will return to the position shown in FIG.","4 when there is no applied voltage thereby obviating the need for a centring spider.","FIG.","6 shows the relationship between rotation and linear travel for the arrangement described with respect to FIGS.","4 and 5.There is a central region having a substantially linear relationship between linear travel and rotation angle, with a tail-off in response at each end of the plot which corresponds to one or other of the springs 2a,2b being in its unstressed state (i.e.","inner and outer rings 4a,4b coplanar).","It will be understood that the above configuration allows small angular displacements to be converted into relatively large linear displacements.","One particular application for the devices of the present invention is in loudspeaker driver units.","FIGS.","7a to 7c are schematic representations of a loudspeaker driver unit in its extreme inner (FIG.","7a), intermediate (FIG.","7b) and extreme outer (FIG.","7c) positions.","The driver unit comprises first and second three-arm springs 2a,2b which are similar to that shown in FIG.","1 with θ=100° and D=17 mm, first and second spiral ceramic piezoelectric actuators 12, 14 (made from a 0.6 mm thick tape of PZT-5H (Morgan Electroceramics) having a lateral piezoelectric strain constant of 274 pC/N, inner spiral diameter 3 mm, outer spiral diameter 10 mm and height 4.5 mm) and a hemisphere front piece 20.The arrangement and mounting of the springs 2a,2b and actuators 12, 14 is as described with reference to the embodiment of FIGS.","4 and 5, with each spring 2a,2b being offset from its planar rest position by 5 mm.","The rim of the hemisphere front piece 20 is secured to the outer ring 4b of the second spring 2b.","The whole assembly is mounted within a cylindrical housing 22 with minimal clearance between the housing 22 and the outer periphery of the outer rings 4a,4b.","The maximum rotation angle from the piezoelectric spiral can be calculated by considering the behaviour of a bimorph tape with a pre-existing curvature.","The change in angle subtended per unit turn of such a tape ΔθN can be calculated from:— Δ θ N = 4 π ɛ R m t where Rm is the mean radius of curvature of the tape at rest, t is the total tape thickness and ε the maximum strain developed in the bimorph tape given by ε=±1.5Ed31 where E is the electric field across each half of the bimorph tape (i.e.","2V/t) where V is the applied voltage, and d31 is the planar coupling coefficient of the piezoelectric material.","The spiral can be considered to be a collection of such curved tapes connected mechanically in series and electrically in parallel.","A close approximation of a spiral geometry can be produced by connecting half turns together in the correct number.","The overall angular actuation, Δθ, can then be given by adding the individual components together:— Δ θ = ∑ 1 n 2 π ɛ R m , n t For the spiral described with reference to FIG.","7, the actuation angles for an applied field of 500V/mm are given in Table 1 below.","The relationship between linear travel and rotation is plotted in FIG.","8, from which it can be seen that peak to peak displacements of +/−3 mm can theoretically be produced.","TABLE 1 Half turn Mean radius, Angular actuation Tape no.","Rm/mm ΔθN/degrees length/mm 1 1.8 0.22 5.7 2 2.2 0.27 6.9 3 2.6 0.32 8.2 4 3.0 0.41 9.4 5 3.4 0.46 10.7 6 3.8 0.51 11.9 7 4.2 0.56 13.2 8 4.6 0.61 14.5 Totals 3.3 80.5 Since the arrangement shown in FIG.","7 has good rigidity, no centring mechanism is required and only a small clearance between the hemisphere driver 20 and the surrounding housing 22 is required to isolate effectively the forward acoustic wave from the equal and opposite acoustic wave.","In a slightly modified arrangement (not shown), a thin rubber flange is provided between the edge of the hemisphere driver and the surrounding housing.","Such a flange seals the unit and provides a centring and balancing force.","Since the overall size of the driver unit is small (<25 mm) the unit can function as a full range audio driver with an essentially flat power response from 20 Hz to 20 kHz, the conventionally stated maximum range of human hearing.","In a working loudspeaker, the whole driver unit is mounted in a suitable cabinet such as would be used for a conventional magnetic driver.","In a further modification (not shown) of the loudspeaker driver unit described, the hemisphere front piece can be replaced by a conical diaphragm.","It will be understood that the described loudspeaker driver unit is light and compact.","In addition, conventional electrodynamic loudspeaker drivers require centring spiders and complicated mounting arrangements so that the coil is held rigidly against radial movement in a strong magnetic field whilst being freely moveable axially.","Such arrangements inevitably impart excessive stiffness to the assembly, causing extra power losses through damping and hysteresis.","Radial movement in the actuator assembly of the present invention is prevented (or at least substantially reduced) due to the inherent high radial stiffness of the plate springs.","Although the arrangement described with reference to FIG.","7 is adequate for most loudspeaker applications, extra stiffening can be provided to prevent tilting of the hemisphere, thereby further improving sound quality.","Such an arrangement is shown in FIGS.","8a to 8c (showing extreme inner, intermediate and extreme outer positions respectively).","Referring to FIG.","8b, the inner rings 6a,6b of the springs 2a,2b are mounted to the respective actuators 12, 14 adjacent one another, and the outer rings 4a,4b are spaced apart on opposite sides thereof.","A cylindrical stiffening collar 24 is adhered at each end to one or other of the outer rings 4a,4b.","Upon actuation both outer rings 4a,4b and the stiffening cylinder 24 move in unison.","FIGS.","9a to 9c show an intermediate arrangement having a shorter stiffening collar 26.Thus arrangement offers a compromise between the compactness of the arrangement of FIGS.","7a to 7c and the stiffness of the arrangement of FIGS.","8a to 8c.","Referring to FIG.","10, a low frequency vibration sensor comprises a pair of springs 2a,2b (identical to those described in relation to FIG.","7) whose outer rings 4a,4b are soldered to a stiffening ring as described in relation to FIG.","4.The outer rings 4a,4b are fixedly mounted to the side walls of a housing 30 so that in use, the plane of the springs is horizontal (as shown in FIG.","10).","The first and second inner rings 6a,6b are offset by an equal amount above and below the outer rings 4a,4b respectively.","The inner ring 6a of the first spring 2a is secured by tabs to the inner curved surface of a spiral transducer 32 (similar to the actuators 12, 14 described with reference to FIG.","7) at its top end, and the inner ring 6b of the second spring 2b is secured by tabs to the outer curved surface of the transducer 32 at its bottom end.","It should be noted that unlike the assembly described with reference to FIG.","7, only a single spiral is required (although two spirals arranged in a similar manner to that described with reference to FIG.","7 would work).","The springs 2a,2b and transducer 32 are sealed in the housing 30 by top and bottom end stops 34.The end stops 34 are positioned at a predetermined distance to limit the maximum vertical travel of the transducer 32, to avoid overstressing the device in use.","The transducer 32 is connected to a voltage meter located externally of the housing 30 by electrodes (not shown).","In use, as the piezoelectric spiral transducer 32 (and inner rings 6a,6b to which it is secured) is forced to move vertically through the inertial forces applied by a vertical vibration (eg.","seismic vibration) through the outer rings 4a,4b, the first and second inner rings 6a,6b will rotate relative to each other, causing the piezoelectric spiral transducer 32 to rotate also.","This rotation generates a charge in the piezoelectric spiral transducer 32 proportional to the degree of linear movement of the inner rings 6a,6b relative to the outer rings 4a,4b.","The stiffness of the spiral arms 10 and the piezoelectric spiral transducer 32 combined, together with the mass of the moveable transducer portion of the system, will cause the device to exhibit a primary resonant frequency.","This resonant frequency can be chosen by altering the geometry of the structure.","For seismic sensing applications, this frequency may be chosen to be as low as 10 Hz.","The frequency range for measurements may then cover the range from 10 Hz up to several hundred Hz.","Since the piezoelectric device does not require a heavy magnet to function, the overall device can be made much lighter than is the case for a corresponding magnet-coil arrangement.","The complexity of the structure can be simplified and the overall size may be reduced for the same response.","Piezoelectric transducers are inherently more efficient than electromagnets, being dependent on electric field rather than current.","At rest in any position, a piezoelectrically driven device draws practically no current, whereas the position of a electromagnet driver is related to the magnitude of constant current drawn.","Thus, the actuator assembly of the present invention is potentially useful in many applications where electromagnetic drivers are used.","The lightweight compact nature of the actuator assembly makes it suitable for any application where mass and size are important considerations.","Similar advantages are obtained by the use of the piezoelectric transducer to generate an electrical signal relative to magnet-moving coil systems."]],"string":"[\n [\n \"Actuator assembly\",\n \"An actuator assembly comprises a first member (6a,6b), a second member (4a,4b) and at least one connecting arm (10a,10b) of fixed length connecting the first and second members (6a,6b;4a,4b).\",\n \"The assembly also comprises an actuator (12,14) which is operably engaged with the first member (6a,6b) so as to be capable of applying a rotating force to the first member (6a,6b).\",\n \"In use, rotational movement of the first member (6a,6b) causes relative linear movement of the first and second members (6a,6b;4a,4b) along an axis.\",\n \"The invention also relates to a loudspeaker driver unit and a sensor incorporating the actuator assembly.\",\n \"In the sensor embodiments, the actuator is an electrical transducer.\"\n ],\n [\n \"1.A method implemented by a push-to-talk wireless mobile terminal for communicating voice information comprising the steps of: determining if a request to send a delayed delivery voice message has been made; if said determining step determines that a request to send a delayed delivery voice message has been made, transmitting an indicator to a communication application server representing an instruction that packets received from the mobile terminal are to be stored for later delivery to a destination Pal; encoding audio input from a user by the mobile terminal into the packets following the determining step; transmitting the packets to the communication application server for later delivery to the destination Pal.\",\n \"2.The method of claim 1 wherein the step of determining if a request to send a delayed delivery voice message has been made comprises sensing that the destination Pal selected by the user to receive the audio input is not available prior to the user initiating the encoding step.\",\n \"3.The method of claim 2 wherein the step of sensing that the Pal selected by the user as the destination party to receive the audio input is not available comprises the step of determining a current status of the selected Pal stored in the mobile terminal, where the status represents that the selected Pal is not available to receive communications.\",\n \"4.The method of claim 1 wherein the step of determining if a request to send a delayed delivery voice message has been made comprises determining that an input signal is entered by the user where the input signal represents a request to send said packets as a delayed delivery voice message regardless of whether or not the destination Pal selected to receive the packets is currently available to receive communications.\",\n \"5.The method of claim 1 wherein the step of transmitting an indicator to a communication application server representing an instruction that the packets are to be stored for later delivery to a destination Pal causes the audio carried by the packets to be stored in the communication application server.\",\n \"6.The method of claim 5 wherein the instruction further conveys to the communication application server that the packets are not to be attempted to be delivered in real-time to the destination Pal.\",\n \"7.The method of claim 1 wherein the step of determining if a request to send a delayed delivery voice message has been made comprises the steps of generating a first request for a real-time voice communication to the destination Pal in response to the push-to-talk button being pressed, providing a first alert to the user indicating that a real-time voice communication to the destination Pal is not available, and sensing a second request to initiate communications to the destination Pal by the push-to-talk button being depressed again following the alert having been provided to the user.\",\n \"8.The method of claim 7 wherein the step of sensing the second request includes sensing the push-to-talk button being depressed again within a predetermined time interval of the first request.\",\n \"9.The method of claim 7 further comprising the step of providing a second alert to the user in response to the second request wherein the second alert comprises a predetermined talk-beep associated with the request to send a delayed delivery voice message, the predetermined talk-beep comprising an audible alert that is different from an audible alert associated with the initiation of a real-time voice communication.\",\n \"10.A push-to-talk wireless mobile terminal for communicating voice information comprising: means for determining if a request to send a delayed delivery voice message has been made; means for transmitting an indicator to a communication application server representing an instruction that packets received from the mobile terminal are to be stored for later delivery to a destination Pal if said determining means determines that a request to send a delayed delivery voice message has been made; means for encoding audio input from a user by the mobile terminal into the packets following the determining step; means for transmitting the packets to the communication application server for later delivery to the destination Pal.\",\n \"11.The mobile terminal of claim 10 wherein the means for determining comprises means for sensing that the destination Pal selected by the user to receive the audio input is not available to receive real-time communications.\",\n \"12.The mobile terminal of claim 11 wherein the means for sensing comprises means for determining a current status of the selected Pal stored in the mobile terminal, where the status represents that the selected Pal is not available to receive communications.\",\n \"13.The mobile terminal of claim 10 wherein the means for determining if a request to send a delayed delivery voice message has been made comprises means for determining that an input signal is entered by the user where the input signal represents a request to send said packets as a delayed delivery voice message regardless of whether or not the destination Pal selected to receive the packets is currently available to receive communications.\",\n \"14.The mobile terminal of claim 10 wherein the means for transmitting an indicator to a communication application server representing an instruction that the packets are to be stored for later delivery to a destination Pal causes the audio carried by the packets to be stored in the communication application server.\",\n \"15.The mobile terminal of claim 14 wherein the instruction transmitted by the means for transmitting further conveys to the communication application server that the packets are not to be attempted to be delivered in real-time to the destination Pal.\",\n \"16.The mobile terminal of claim 10 wherein the means for determining if a request to send a delayed delivery voice message has been made comprises means for generating a first request for a real-time voice communication to the destination Pal in response to the push-to-talk button being pressed, means for providing a first alert to the user indicating that a real-time voice communication to the destination Pal is not available, and means for sensing a second request to initiate communications to the destination Pal by the push-to-talk button being depressed again following the alert having been provided to the user.\",\n \"17.The mobile terminal of claim 16 wherein the means for sensing the second request includes means for sensing the push-to-talk button being depressed again within a predetermined time interval of the first request.\",\n \"18.The mobile terminal of claim 16 further comprising means for providing a second alert to the user in response to the second request, the second alert comprises a predetermined talk-beep associated with the request to send a delayed delivery voice message, the predetermined talk-beep comprising an audible alert that is different from an audible alert associated with the initiation of a real-time voice communication.\",\n \"19.A method implemented by a push-to-talk wireless mobile terminal for communicating voice information comprising the steps of: displaying a list of Pals of the first user including visual indicia representing that a previously transmitted voice message by the first user to a first Pal is stored and awaits delivery to the first Pal; determining if a request has been made by the first user to access the stored voice message; if said request is determined to have been made, discerning the type of access desired by the first user; transmitting a command to a communication application server at which the voice messages stored where the command conveys instructions to the communication application server to implement action based on the type of access desired by the first user.\",\n \"20.The method of claim 19 wherein the step of discerning comprises discerning that the stored voice message is to be played back to the first user and wherein the transmitted command conveys instructions to the communication application server to implement transmission of the stored voice message to the first user, the method further comprising the step of receiving at the mobile terminal of the first user the stored voice message previously transmitted by the first user to the first Pal and audibly playing the stored voice message.\",\n \"21.The method of claim 19 wherein the step of discerning comprises discerning that the stored voice message is to be deleted and wherein the transmitted command conveys instructions to the communication application server to delete the stored voice message previously transmitted by the first user to the first Pal, the method further comprising the step of receiving at the mobile terminal of the first user a signal technology that the stored voice message has been deleted.\",\n \"22.A push-to-talk wireless mobile terminal for communicating voice information comprising: means for displaying a list of Pals of the first user including visual indicia representing that a previously transmitted voice message by the first user to a first Pal is stored and awaits delivery to the first Pal; means for determining if a request has been made by the first user to access the stored voice message; means for discerning the type of access desired by the first user if said request is determined to have been made; means for transmitting a command to a communication application server at which the voice messages are stored where the command conveys instructions to the communication application server to implement action based on the type of access desired by the first user.\",\n \"23.The mobile terminal of claim 22 wherein the means for discerning discerns that the stored voice message is to be played back to the first user and wherein the means for transmitting transmits the command that conveys instructions to the communication application server to implement transmission of the stored voice message to the first user, the mobile terminal further comprising means for receiving, at the mobile terminal of the first user, the stored voice message previously transmitted by the first user to the first Pal and audibly playing the stored voice message.\",\n \"24.The mobile terminal of claim 22 wherein the means for discerning discerns that the stored voice message is to be deleted and wherein the means for transmitting transmits the command that conveys instructions to the communication application server to delete the stored voice message previously transmitted by the first user to the first Pal, the mobile terminal further comprising means for receiving at the mobile terminal of the first user a signal representing that the stored voice message has been deleted.\",\n \"25.A method implemented by a push-to-talk wireless mobile terminal for communicating voice information comprising the steps of: displaying a list of Pals including visual indicia of whether a voice message is waiting for delivery from a Pal; determining if a request to receive a waiting voice message has been initiated by a user of the mobile terminal; if said request is determined to have been made, transmitting at least one packet to a communication application server requesting that the waiting voice message associated with a selected Pal be delivered to the user's mobile terminal; receiving packets at the user's mobile terminal from the communication application server containing the waiting voice message and playing the message to the user.\",\n \"26.The method of claim 25 wherein the steps of determining if the request has been made comprises sensing that a Pal is selected by the user where the Pal has corresponding visual indicia indicating that a voice message from the Pal is waiting delivery to the user, and sensing an input initiated by the user representing a request to receive delivery of the voice message corresponding to the selected Pal.\",\n \"27.The method of claim 26 further comprising the steps of receiving a status update following receiving the packets where the status update indicates that there is no longer the voice message from the Pal waiting delivery to the user and updating the visual indicia corresponding to the Pal whose voice message was received to show another visual indicia representing that the voice message is no longer waiting delivery to the user.\",\n \"28.A push-to-talk wireless mobile terminal for communicating voice information comprising: means for displaying a list of Pals including visual indicia of whether a voice message is waiting for delivery from a Pal; means for determining if a request to receive a waiting voice message has been initiated by a user of the mobile terminal; means for transmitting at least one packet to a communication application server requesting that the waiting voice message associated with a selected Pal be delivered to the user's mobile terminal if said request is determined to have been made; means for receiving packets at the user's mobile terminal from the communication application server containing the waiting voice message and playing the message to the user.\",\n \"29.The mobile terminal of claim 28 wherein the means for determining if the request has been made comprises means for sensing that a Pal is selected by the user where the Pal has corresponding visual indicia indicating that a voice message from the Pal is waiting delivery to the user, and means for sensing an input initiated by the user representing a request to receive delivery of the voice message corresponding to the selected Pal.\",\n \"30.The mobile terminal of claim 29 further comprising means for receiving a status update following receiving the packets where the status update indicates that there is no longer the voice message from the Pal waiting delivery to the user and means for updating the visual indicia corresponding to the Pal whose voice message was received to show another visual indicia representing that the voice message is no longer waiting delivery to the user.\",\n \"31.A method implemented by a communication application server in a packet communication network for processing communications comprising the steps of: receiving a first packet from a mobile terminal of a user; determining if the first packet contains an indicator representing an instruction to process any following voice packets as a delayed delivery voice message; receiving a set of voice packets from a mobile terminal of a user; if said determining step determines that said indicator is present, storing at least payloads of the voice packets of said set in memory and labeling the stored payloads as addressed to a destination Pal identified by said set of packets.\",\n \"32.The method of claim 31 further comprising the step of attempting to deliver voice information contained in said payloads only upon receiving a command from a mobile terminal of the destination Pal where the command corresponds to input initiated by the Pal to retrieve the stored voice information.\",\n \"32.The method of claim 31 further comprising the steps of generating a status update following the storing step and transmitting the status update to at least the mobile terminals of the destination Pal and the user.\",\n \"33.The method of claim 32 wherein the step of transmitting the status update comprises transmitting a status update to the destination Pal indicating that the voice information from the user is awaiting deliver to the destination Pal and transmitting a status update to the user indicating that the voice information from the user to the destination Pal is still awaiting delivery to the destination Pal.\",\n \"34.The method of claim 32 further comprising the step of receiving said command from the mobile terminal of the destination Pal, retrieving the at least voice information payloads, encoding the at least voice information payloads into further packets addressed to the destination Pal, and transmitting the further packets to the destination Pal.\",\n \"35.The method of claim 34 further comprising the steps of generating a status update following the transmitting of the further packets and transmitting the status update to at least the mobile terminals of the destination Pal and the user, where the status update represents that the voice information has been delivered to the destination Pal.\",\n \"36.The method of claim 35 wherein the step of transmitting the status update comprises transmitting a status update to the destination Pal indicating that there is no longer the voice information from the user awaiting deliver to the destination Pal and transmitting a status update to the user indicating that there is no longer the voice information from the user awaiting delivery to the destination Pal.\",\n \"37.A communication application server in a packet communication network for processing communications comprising: means for receiving a first packet from a mobile terminal of a user; means for determining if the first packet contains an indicator representing an instruction to process any following voice packets as a delayed delivery voice message; means for receiving a set of voice packets from a mobile terminal of a user; means for storing at least payloads of the voice packets of said set in memory and labeling the stored payloads as addressed to a destination Pal identified by said set of packets if said determining step determines that said indicator is present.\",\n \"38.The communication application server of claim 37 further comprising means for attempting to deliver voice information contained in said payloads only upon receiving a command from a mobile terminal of the destination Pal where the command corresponds to input initiated by the Pal to retrieve the stored voice information.\",\n \"39.The communication application server of claim 37 further comprising means for generating a status update following the storage of the at least payloads and means for transmitting the status update to at least the mobile terminals of the destination Pal and the user.\",\n \"40.The communication application server of claim 39 wherein the means for transmitting the status update comprises means for transmitting a status update to the destination Pal indicating that the voice information from the user is awaiting delivery to the destination Pal and means for transmitting a status update to the user indicating that the voice information from the user to the destination Pal is still awaiting delivery to the destination Pal.\",\n \"41.The communication application server of claim 39 further comprising means for receiving said command from the mobile terminal of the destination Pal, means for retrieving the at least voice information payloads, means for encoding the at least voice information payloads into further packets addressed to the destination Pal, and means for transmitting the further packets to the destination Pal.\",\n \"42.The communication application server of claim 41 further comprising means for generating a status update following the transmitting of the further packets and means for transmitting the status update to at least the mobile terminals of the destination Pal and the user, where the status update represents that the voice information has been delivered to the destination Pal.\",\n \"43.The communication application server of claim 42 wherein the means for transmitting the status update comprises means for transmitting a status update to the destination Pal indicating that there is no longer the voice information from the user awaiting deliver to the destination Pal and means for transmitting a status update to the user indicating that there is no longer the voice information from the user awaiting delivery to the destination Pal.\"\n ],\n [\n \"The present invention relates, in a first aspect, to an actuator assembly in which a rotating force applied to a first member is converted into linear movement.\",\n \"The present invention also relates to loudspeaker driver units comprising such a device.\",\n \"There are many devices (such as loudspeaker driver units and focussing assemblies for cameras and microscopes etc and CD and DVD players) into which linear actuator assemblies are incorporated.\",\n \"In such devices one or more of the following factors may be important: mass, power consumption, compactness and linearity of response.\",\n \"For example, in magnet-based loudspeaker driver units, a moving coil oscillates in a magnetic field generated by a permanent magnet.\",\n \"The magnet may make up as much as 90% of the mass of the driver unit, but serves no other purpose than to generate the magnetic field.\",\n \"Crystal loudspeaker driver units are also known.\",\n \"These incorporate a piezoelectric crystal which undergoes a change in thickness when subjected to a potential difference.\",\n \"When an alternating voltage is applied, the crystal undergoes bending oscillations which are transmitted to a speaker diaphragm.\",\n \"Crystal-based driver units are particularly suited to the generation of high frequencies, and a conventional loudspeaker may comprise an electrodynamic- and crystal-based driver unit in order to achieve a full range (20 Hz-20 kHz) frequency response.\",\n \"Geophone sensors (see for example U.S. Pat.\",\n \"No.\",\n \"4,152,692), used to detect seismic vibrations, basically comprise a coil suspended between a pair of springs and positioned around a strong magnet.\",\n \"Vertical linear travel of the coil due to seismic vibrations produces strong electrical signals in the coil.\",\n \"Such magnet-coil based devices are generally complex and relatively heavy and large.\",\n \"An object of a first aspect of the present invention is to provide a novel linear actuator assembly which obviates or mitigates one or more disadvantages of known actuator assemblies.\",\n \"A further object of the present invention is to provide improved devices, such as a loudspeaker driver unit, incorporating such an assembly.\",\n \"An object of a second aspect of the present invention is to provide an improved vibration sensor.\",\n \"According to the present invention, there is provided an actuator assembly comprising:— (i) a first member, (ii) a second member, (iii) at least one connecting arm of fixed length connecting said first and second members, and (iv) an actuator operably engaged with said first member so as to be capable of applying a rotating force to the first member, wherein in use, rotational movement of the first member causes relative linear movement of the first and second members along an axis.\",\n \"Preferably, the first member and actuator are arranged so that the force applied by the latter causes the former to rotate in a plane substantially perpendicular to said axis.\",\n \"The actuator is preferably a piezoelectric or electrostrictive transducer.\",\n \"Preferably, the actuator is in the form of a spiral having at least one half turn.\",\n \"It will be understood that the greater the number of turns for a given actuator, the greater the maximum angular actuation will be.\",\n \"Examples of suitable piezoelectric materials include ceramic materials such as lead-zirconate-titanate (PZT) based systems or non-ceramic systems (eg.\",\n \"polymer based systems such as polyvinylidene fluoride).\",\n \"Particularly preferred compositions are those classified by the US Department of Defence under DOD STD-1376A type VI.\",\n \"An example of which is PZT-5H (sold by Morgan Electroceramics).\",\n \"Preferably, the piezoelectric material has a lateral piezoelectric strain (d31) coefficient greater than 200 pC/N.\",\n \"More preferably the d3coefficient is no more than 350 pC/N.\",\n \"Preferably the elastic stiffness of the piezoelectric material is at least 65 GPa.\",\n \"When a piezoelectric material is used it preferably has a bimorph or multimorph structure, although unimorph structures may be used.\",\n \"If an electrostrictive material is used, it is preferably a ceramic material, and more preferably based on the lead magnesium niobate-lead titanate (PMN-PT) system.\",\n \"Preferably, the first and second members are annular with differing diameters (“inner” and “outer” annular members) interconnected by at least two (but preferably three) connecting arms.\",\n \"Preferably, said arms are arranged symmetrically between said annular members.\",\n \"More preferably, said arms are arcuate.\",\n \"The first member may be the inner or outer member.\",\n \"Preferably, the first and second members and said at least one connecting arm are of unitary construction.\",\n \"Such a construction in which inner and outer annular rings are interconnected by arcuate connecting arms will hereinafter be referred to as a “plate spring” or a “spiral arm spring”.\",\n \"Such springs are per se known and have been used in geophone sensor units.\",\n \"It will be understood that when the inner and outer rings of a plate spring are moved apart along an axis perpendicular to the plane of the spring, there must be relative rotation of the inner and outer rings because the connecting arms are of fixed length.\",\n \"Consequently, if one ring is caused to rotate and the other is prevented from rotating, relative linear movement of the rings will be induced perpendicular to the plane of the spring.\",\n \"It will also be understood from the foregoing that one of the first and second members may be mounted so as to prevent movement along the axis, in which case actuation will result in movement of the other member along the axis.\",\n \"In a first series of embodiments, the first member is mounted so that linear movement along the axis is prevented, whereas the second member is mounted for linear movement along the axis.\",\n \"In a preferred arrangement, the actuator (preferably in spiral form) is positioned inside the first (inner) annular member and secured thereto.\",\n \"Securement may be achieved by, for example, soldering, fusing, or bonding with adhesive.\",\n \"Suitable formations (eg.\",\n \"tabs or flanges) onto which to secure the actuator may be provided on the first member.\",\n \"Alternatively, the actuator (preferably in spiral form) is positioned outside the first (outer) annular member and secured thereto.\",\n \"Such an arrangement permits the mounting of, for example, a lens inside the second (inner) annular member.\",\n \"In a highly preferred embodiment, the actuator assembly comprises first and second plate springs whose outer annular rings are secured together (directly or indirectly, eg.\",\n \"by placing a stiffening ring or cylindrical collar therebetween), and first and second spiral actuators arranged to actuate the respective inner rings of the first and second plate springs wherein the actuators are oppositely orientated so that, in use, actuation of the first spiral actuator rotates the inner ring of the first plate spring in one direction about the axis, whereas simultaneous actuation of the second actuator rotates the inner ring of the second plate spring in the opposite direction about the axis by an equal amount, whereby to move the outer rings along the axis.\",\n \"It will be readily understood that the outer rings can be moved in either direction along the axis depending on the polarity of the applied voltage.\",\n \"In a very highly preferred embodiment, the outer rings are secured together and the inner rings are equidistantly spaced either side of the outer rings.\",\n \"In a second series of embodiments, the second member is mounted so that linear movement along the axis is prevented, whereas the first member and actuator are mounted for linear movement along the axis.\",\n \"The present invention also resides in a loudspeaker driver unit and loudspeaker comprising an actuator assembly in accordance with the present invention and an air piston driven by the actuator assembly to generate an acoustic wave.\",\n \"The air piston may be in the form of a hemisphere or a conical diaphragm.\",\n \"Alternatively, the loudspeaker driver unit may include a diaphragm which is oscillated by the actuator assembly to generate an acoustic wave.\",\n \"It will be understood that if the actuator is in fact an electrical transducer, rotation of the transducer will result in the generation of an electrical signal in the transducer.\",\n \"Thus, according to a second aspect of the present invention, there is provided a sensor comprising:— (i) a first member, (ii) a second member, (iii) at least one connecting arm of fixed length connecting said first and second members and (iv) a transducer operably engaged with said first member, wherein in use, relative linear movement of the first and second members along an axis causes rotational movement of the first member which is transmitted to said transducer whereby to generate an electrical signal in said transducer.\",\n \"Said sensor corresponds closely to said actuator assembly, the primary difference being that in the former, relative linear movement is converted into rotational movement and subsequently into an electrical signal, whereas in the latter, actuated rotational movement is converted into relative linear movement.\",\n \"It will therefore be understood that the preferred features of the assembly of the first aspect are also preferred features of the sensor of the second aspect.\",\n \"The sensor may be a vibration sensor, e.g.\",\n \"for detecting seismic vibrations.\",\n \"Embodiments of the present invention will now be described by way of example only, with reference to the accompanying drawings in which:— FIG.\",\n \"1 is plan view of a plate spring suitable for use in the actuator assembly of the first aspect of the invention, or the sensor of the second aspect of the invention, FIG.\",\n \"2 shows the triangle formed between an arm of length L lying on a diameter D when the spring of FIG.\",\n \"1 is offset by a displacement 6, FIG.\",\n \"3 is a graph of linear travel against relative rotation derived for the spring of FIG.\",\n \"1, FIGS.\",\n \"4 and 5 show an embodiment of an actuator assembly in accordance with the first aspect of the present invention, FIG.\",\n \"6 is a graph of linear travel against relative rotation derived for the embodiment of FIGS.\",\n \"4 and 5.FIGS.\",\n \"7a to 7c are schematic representations of a loudspeaker driver unit incorporating an actuator assembly in accordance with the first aspect of the present invention in an extreme inner (FIG.\",\n \"7a), intermediate (FIG.\",\n \"7b) and extreme outer (FIG.\",\n \"7c) position, FIGS.\",\n \"8a to 8c correspond to FIGS.\",\n \"7a to 7c for a stiffened loudspeaker driver unit incorporating an actuator assembly in accordance with the first aspect of the present invention, FIGS.\",\n \"9a to 9c correspond to FIGS.\",\n \"7a to 7c for a partially stiffened loudspeaker driver unit incorporating an actuator assembly in accordance with the first aspect of the present invention, and FIG.\",\n \"10 is a schematic view of part of a vibration sensor in accordance with the second aspect of the invention.\",\n \"Referring to FIG.\",\n \"1, a plate spring 2 comprises a first (outer) annular ring 4 and a second (inner) annular ring 6, the first and second rings 4,6 being concentric (i.e.\",\n \"coplanar) in the rest position of the spring 2.A pair of tabs 8 angularly spaced by 180° extends radially inwardly from the inner ring 6.The first and second rings are connected by three part-annular connecting arms 10.Each arm 10 subtends an angle of 0 and lies on a circle of diameter D. The plate spring 2 is of unitary construction and fabricated from beryllium copper alloy.\",\n \"In use, the tabs 8 are bent out of the plane of the spring 2 and serve as mounting points for a spiral actuator (described below).\",\n \"It will be understood that as the two rings 4,6 are moved relative to one another along an axis perpendicular to the plane of the spring 2, there must be relative rotation of the rings 4,6 to maintain the rings 4,6 in a parallel orientation.\",\n \"This is because the connecting arms 10 are of fixed length.\",\n \"An approximate relationship between the linear movement and the relative rotation can be derived as follows:— The length of each arm 10 is given by L = D θ 2 If the inner and outer rings 4,6 are displaced perpendicularly to the plane of the spring 2 relative to each other by a distance δ, the angle subtended by each arm 10 (as viewed in FIG.\",\n \"1) must change.\",\n \"This can be calculated approximately by assuming that the arms 10 lie on the same diameter (D) and form perfect helical lines connecting the inner and outer rings 4,6.FIG.\",\n \"2 shows the triangle formed by the arc length L and the displacement 6.The projection of the arc on to the initial plane of the spring 2 is then given by L = D 2 ( θ - Δ θ ) where Δθ corresponds to the relative rotation of the inner and outer rings 4,6 of the spring 2 necessary for them to remain parallel, with L being constant.\",\n \"The displacement δ can then be given as θ = D 2 Δ θ 2 + 2 θ Δ θ This relationship is depicted in FIG.\",\n \"3 from which it will be noted that at relatively large linear displacements, the relationship between linear travel and rotation is approximately linear.\",\n \"The present invention takes advantage of this effect which allows linear movement to be produced directly from a rotational actuation.\",\n \"Referring to FIGS.\",\n \"4 and 5, an embodiment of the actuator assembly comprises first and second identical plate springs 2a,2b, first and second identical piezoelectric ceramic actuators 12, 14 and a mounting post 16.The plate springs 2a,2b are similar to that described with reference to FIG.\",\n \"1 (and the same reference numerals are used to denote corresponding structures, suffixed by “a” and “b” to denote the first and second springs respectively), but there are only two connecting arms 10.Each actuator 12, 14 is formed from a tape of a lead-zirconate-titanate (PZT) composition having a bimorph structure which is wound into a spiral having 4 turns in the present embodiment.\",\n \"Such piezoelectric ceramic materials are particularly suited to the present invention because they can exhibit a lateral piezoelectric strain (d31) coefficient as high as 350 pC/N, while possessing a flexural elastic modulus of over 60 Gpa.\",\n \"If only small actuation movements are required, these properties allow high forces to be generated from a small amount of material.\",\n \"This is useful in certain applications, such as in loudspeaker driver units as will be described below.\",\n \"The outer diameter of the actuator spirals 12, 14 corresponds to the inner diameter of the inner rings 6a,6b of the plate springs 2a,2b.\",\n \"A Bimorph piezoelectric structure is formed from two layers of piezoelectric material, separated by a conductive central electrode.\",\n \"Electrodes are placed on the outer surfaces of the ceramic layers, and the layers are poled and actuated using these three electrodes such that the overall effect of the actuation is to expand one ceramic layer while causing the other to contract, through the effect of the d31 coefficient, thus producing a uniform bending strain in the element.\",\n \"Various methods for creating ceramic compositions suitable for use in the present invention are known, see for example EP01 83453, EP0288208 and N. Alford et al, Nature; v.330; pp 51-53.To form the required spiral actuator structure, it is most beneficial to first create the required Bimorph structure in a planar form.\",\n \"This may be done through the routes of printing and lamination.\",\n \"A green (unfired) ceramic tape is formed from PZT powder mixed with a polyvinyl butyral (PVB) binder and cyclohexanone solvent.\",\n \"In the present embodiment the formulation is 100 parts by weight of PZT to 6 parts PVB, to 7 parts cyclohexanone and 0.1 parts stearic acid, the stearic acid serving as a surfactant.\",\n \"The green tape is then printed with the internal electrode, which may be of platinum, silver or an alloy of silver and palladium, formed into a printable ink.\",\n \"In the present embodiment platinum is used (grade C51121D1 supplied by Gwent Electronic Materials, Pontypool).\",\n \"The printed tape is then laminated with another ceramic tape of the same type and thickness (in the present embodiment PZT-5H, each tape 0.35 mm thick in the green state).\",\n \"The lamination step may involve pressure and/or heat to achieve a strong bond across the electrode print.\",\n \"The outer electrodes are then printed in the same fashion as the internal electrode, and allowed to dry.\",\n \"After the printing stage, the overall tape structure must be sufficiently flexible and plastic to be deformed into the required spiral actuator structure.\",\n \"This shaping may be achieved by using a tape formation route which includes a thermoplastic binder, in which case heat and pressure may be used to deform the tape into the required spiral.\",\n \"With a solvent and binder system as in the present embodiment, the presence of the solvent allows the material to remain plastically deformable prior to removal of the solvent through evaporation.\",\n \"An interleaving tape may be used, in order to maintain separation of the spiral turns during shaping.\",\n \"This material may be in the form of carbon, formed in the same manner as the ceramic tape.\",\n \"In the present embodiment 35 parts by weight of carbon black, 11 parts PVB and 12 parts cyclohexanone are used.\",\n \"After the plastic processing step, and after drying, if applicable, the carbon tape is removed along with the binder in the ceramic tape through slow heating up to 600° C. in air.\",\n \"Extra interleaving layers may be used between the PZT and the carbon layers to prevent the tapes adhering to each other while the solvent is still present.\",\n \"Suitable materials include polythene, preferably less than 50:m thick.\",\n \"This may be removed from the spirals after drying.\",\n \"The spiral form is then sintered in an enclosed crucible with sufficient excess PbO-containing material, such as lead zirconate, to prevent PbO loss from the piezoelectric material.\",\n \"After sintering, the thickness of the tapes is reduced to about 0.3 mm giving a total actuator thickness of 0.6 mm.\",\n \"Soldered electrical connections are then made to the three separate electrode layers, with a wire connected to each.\",\n \"The outer two layers of the tape are connected to a high voltage supply, and the device is placed in a heated oil bath at 120-130° C. A voltage equivalent to 2.5 kV/mm across the whole tape thickness is applied while the device is in the bath for 10 minutes.\",\n \"This process polarises the piezoelectric tape.\",\n \"After polarisation, the third electrode, connected to the central electrode layer, can be used to apply a field which is in opposite directions on each half of the tape.\",\n \"The outer two electrodes can therefore be connected together, and used as the ground electrode, while the central electrode can be used for the driving signal.\",\n \"For driving, the opposing electric fields generate bending in the tape.\",\n \"Each of the actuator spirals 12, 14 is securely mounted onto the mounting post 16, each actuator 12, 14 being oppositely orientated relative to the other with the actuators 12, 14 spaced a short distance apart.\",\n \"The outer rings 4a,4b of the springs 2a,2b are securely soldered to opposite sides of a stiffening ring (not shown) which prevents warping of the outer rings 4a,4b.\",\n \"It should be noted that the springs 2a,2b are in the same orientation.\",\n \"Each spring 2a,2b is tensioned by moving the inner ring 6a,6b out of the initial plane of the spring 2a,2b.\",\n \"The inner ring 6a of the first spring 2a is moved towards the end of the first actuator 12 remote from the second actuator 14 where it is securely fixed via the tabs (8, FIG.\",\n \"1) to the outer curved surface of the first actuator 12 by soldered joints.\",\n \"The tabs lie 180° apart so that the axial forces on the outer rings 4a,4b do not produce unbalanced forces on the actuator spirals 12, 14.The inner ring 6b of the second spring 2b is moved in the opposite direction (i.e.\",\n \"towards the end of the second actuator 14 remote from the first actuator 12) where it is secured to the curved surface of the second actuator 14 via the tabs (8, FIG.\",\n \"1) to the outer curved surface of the second actuator 14 by soldered joints.\",\n \"As can be seen in FIG.\",\n \"4, the soldered outer rings 4a,4b of the springs 2a,2b are equidistant from the respective inner rings 6a,6b.\",\n \"In use, when an electrical signal is applied to both actuator spirals 12,14 in parallel via the respective inner and outer electrodes connected to a power source (not shown), the actuators 12, 14 “rotate” in opposite directions.\",\n \"The rotation of the first actuator 12 is transmitted to the inner ring 6a of the first spring 2a and the opposite rotation of the second actuator 14 is transmitted to the inner ring 6b of the second spring 2b.\",\n \"Since the inner rings 6a,6b of the springs 2a,2b are prevented from translational movement by means of their securement to the respective actuator 12, 14, the outer rings 4a,4b move along an axis perpendicular to the planes of the springs 2a,2b.\",\n \"This is the only way in which the rotational force can be transmitted, since the first and second outer rings 4a,4b, although capable of rotation, are subjected to equal but opposite rotational forces from the respective actuators 12, 14.No rotation results because of their securement to the stiffening ring.\",\n \"Thus, it will be understood that the two actuators 12, 14 work in unison.\",\n \"Depending on the overall design of the device, the travel of the outer rings 4a,4b may be anything from about 100 μm up to several centimetres.\",\n \"If the polarity of the applied voltage is reversed, the outer rings 4a,4b will move along the axis in the opposite direction.\",\n \"The outer rings 4a,4b can be moved between the respective planes of the inner rings 6a,6b of the first and second springs 2a,2b, the exact position of the outer rings 4a,4b being dependent on the polarity and magnitude of the applied voltage to the actuators 12, 14.The actuator assembly is self centring, i.e.\",\n \"will return to the position shown in FIG.\",\n \"4 when there is no applied voltage thereby obviating the need for a centring spider.\",\n \"FIG.\",\n \"6 shows the relationship between rotation and linear travel for the arrangement described with respect to FIGS.\",\n \"4 and 5.There is a central region having a substantially linear relationship between linear travel and rotation angle, with a tail-off in response at each end of the plot which corresponds to one or other of the springs 2a,2b being in its unstressed state (i.e.\",\n \"inner and outer rings 4a,4b coplanar).\",\n \"It will be understood that the above configuration allows small angular displacements to be converted into relatively large linear displacements.\",\n \"One particular application for the devices of the present invention is in loudspeaker driver units.\",\n \"FIGS.\",\n \"7a to 7c are schematic representations of a loudspeaker driver unit in its extreme inner (FIG.\",\n \"7a), intermediate (FIG.\",\n \"7b) and extreme outer (FIG.\",\n \"7c) positions.\",\n \"The driver unit comprises first and second three-arm springs 2a,2b which are similar to that shown in FIG.\",\n \"1 with θ=100° and D=17 mm, first and second spiral ceramic piezoelectric actuators 12, 14 (made from a 0.6 mm thick tape of PZT-5H (Morgan Electroceramics) having a lateral piezoelectric strain constant of 274 pC/N, inner spiral diameter 3 mm, outer spiral diameter 10 mm and height 4.5 mm) and a hemisphere front piece 20.The arrangement and mounting of the springs 2a,2b and actuators 12, 14 is as described with reference to the embodiment of FIGS.\",\n \"4 and 5, with each spring 2a,2b being offset from its planar rest position by 5 mm.\",\n \"The rim of the hemisphere front piece 20 is secured to the outer ring 4b of the second spring 2b.\",\n \"The whole assembly is mounted within a cylindrical housing 22 with minimal clearance between the housing 22 and the outer periphery of the outer rings 4a,4b.\",\n \"The maximum rotation angle from the piezoelectric spiral can be calculated by considering the behaviour of a bimorph tape with a pre-existing curvature.\",\n \"The change in angle subtended per unit turn of such a tape ΔθN can be calculated from:— Δ θ N = 4 π ɛ R m t where Rm is the mean radius of curvature of the tape at rest, t is the total tape thickness and ε the maximum strain developed in the bimorph tape given by ε=±1.5Ed31 where E is the electric field across each half of the bimorph tape (i.e.\",\n \"2V/t) where V is the applied voltage, and d31 is the planar coupling coefficient of the piezoelectric material.\",\n \"The spiral can be considered to be a collection of such curved tapes connected mechanically in series and electrically in parallel.\",\n \"A close approximation of a spiral geometry can be produced by connecting half turns together in the correct number.\",\n \"The overall angular actuation, Δθ, can then be given by adding the individual components together:— Δ θ = ∑ 1 n 2 π ɛ R m , n t For the spiral described with reference to FIG.\",\n \"7, the actuation angles for an applied field of 500V/mm are given in Table 1 below.\",\n \"The relationship between linear travel and rotation is plotted in FIG.\",\n \"8, from which it can be seen that peak to peak displacements of +/−3 mm can theoretically be produced.\",\n \"TABLE 1 Half turn Mean radius, Angular actuation Tape no.\",\n \"Rm/mm ΔθN/degrees length/mm 1 1.8 0.22 5.7 2 2.2 0.27 6.9 3 2.6 0.32 8.2 4 3.0 0.41 9.4 5 3.4 0.46 10.7 6 3.8 0.51 11.9 7 4.2 0.56 13.2 8 4.6 0.61 14.5 Totals 3.3 80.5 Since the arrangement shown in FIG.\",\n \"7 has good rigidity, no centring mechanism is required and only a small clearance between the hemisphere driver 20 and the surrounding housing 22 is required to isolate effectively the forward acoustic wave from the equal and opposite acoustic wave.\",\n \"In a slightly modified arrangement (not shown), a thin rubber flange is provided between the edge of the hemisphere driver and the surrounding housing.\",\n \"Such a flange seals the unit and provides a centring and balancing force.\",\n \"Since the overall size of the driver unit is small (<25 mm) the unit can function as a full range audio driver with an essentially flat power response from 20 Hz to 20 kHz, the conventionally stated maximum range of human hearing.\",\n \"In a working loudspeaker, the whole driver unit is mounted in a suitable cabinet such as would be used for a conventional magnetic driver.\",\n \"In a further modification (not shown) of the loudspeaker driver unit described, the hemisphere front piece can be replaced by a conical diaphragm.\",\n \"It will be understood that the described loudspeaker driver unit is light and compact.\",\n \"In addition, conventional electrodynamic loudspeaker drivers require centring spiders and complicated mounting arrangements so that the coil is held rigidly against radial movement in a strong magnetic field whilst being freely moveable axially.\",\n \"Such arrangements inevitably impart excessive stiffness to the assembly, causing extra power losses through damping and hysteresis.\",\n \"Radial movement in the actuator assembly of the present invention is prevented (or at least substantially reduced) due to the inherent high radial stiffness of the plate springs.\",\n \"Although the arrangement described with reference to FIG.\",\n \"7 is adequate for most loudspeaker applications, extra stiffening can be provided to prevent tilting of the hemisphere, thereby further improving sound quality.\",\n \"Such an arrangement is shown in FIGS.\",\n \"8a to 8c (showing extreme inner, intermediate and extreme outer positions respectively).\",\n \"Referring to FIG.\",\n \"8b, the inner rings 6a,6b of the springs 2a,2b are mounted to the respective actuators 12, 14 adjacent one another, and the outer rings 4a,4b are spaced apart on opposite sides thereof.\",\n \"A cylindrical stiffening collar 24 is adhered at each end to one or other of the outer rings 4a,4b.\",\n \"Upon actuation both outer rings 4a,4b and the stiffening cylinder 24 move in unison.\",\n \"FIGS.\",\n \"9a to 9c show an intermediate arrangement having a shorter stiffening collar 26.Thus arrangement offers a compromise between the compactness of the arrangement of FIGS.\",\n \"7a to 7c and the stiffness of the arrangement of FIGS.\",\n \"8a to 8c.\",\n \"Referring to FIG.\",\n \"10, a low frequency vibration sensor comprises a pair of springs 2a,2b (identical to those described in relation to FIG.\",\n \"7) whose outer rings 4a,4b are soldered to a stiffening ring as described in relation to FIG.\",\n \"4.The outer rings 4a,4b are fixedly mounted to the side walls of a housing 30 so that in use, the plane of the springs is horizontal (as shown in FIG.\",\n \"10).\",\n \"The first and second inner rings 6a,6b are offset by an equal amount above and below the outer rings 4a,4b respectively.\",\n \"The inner ring 6a of the first spring 2a is secured by tabs to the inner curved surface of a spiral transducer 32 (similar to the actuators 12, 14 described with reference to FIG.\",\n \"7) at its top end, and the inner ring 6b of the second spring 2b is secured by tabs to the outer curved surface of the transducer 32 at its bottom end.\",\n \"It should be noted that unlike the assembly described with reference to FIG.\",\n \"7, only a single spiral is required (although two spirals arranged in a similar manner to that described with reference to FIG.\",\n \"7 would work).\",\n \"The springs 2a,2b and transducer 32 are sealed in the housing 30 by top and bottom end stops 34.The end stops 34 are positioned at a predetermined distance to limit the maximum vertical travel of the transducer 32, to avoid overstressing the device in use.\",\n \"The transducer 32 is connected to a voltage meter located externally of the housing 30 by electrodes (not shown).\",\n \"In use, as the piezoelectric spiral transducer 32 (and inner rings 6a,6b to which it is secured) is forced to move vertically through the inertial forces applied by a vertical vibration (eg.\",\n \"seismic vibration) through the outer rings 4a,4b, the first and second inner rings 6a,6b will rotate relative to each other, causing the piezoelectric spiral transducer 32 to rotate also.\",\n \"This rotation generates a charge in the piezoelectric spiral transducer 32 proportional to the degree of linear movement of the inner rings 6a,6b relative to the outer rings 4a,4b.\",\n \"The stiffness of the spiral arms 10 and the piezoelectric spiral transducer 32 combined, together with the mass of the moveable transducer portion of the system, will cause the device to exhibit a primary resonant frequency.\",\n \"This resonant frequency can be chosen by altering the geometry of the structure.\",\n \"For seismic sensing applications, this frequency may be chosen to be as low as 10 Hz.\",\n \"The frequency range for measurements may then cover the range from 10 Hz up to several hundred Hz.\",\n \"Since the piezoelectric device does not require a heavy magnet to function, the overall device can be made much lighter than is the case for a corresponding magnet-coil arrangement.\",\n \"The complexity of the structure can be simplified and the overall size may be reduced for the same response.\",\n \"Piezoelectric transducers are inherently more efficient than electromagnets, being dependent on electric field rather than current.\",\n \"At rest in any position, a piezoelectrically driven device draws practically no current, whereas the position of a electromagnet driver is related to the magnitude of constant current drawn.\",\n \"Thus, the actuator assembly of the present invention is potentially useful in many applications where electromagnetic drivers are used.\",\n \"The lightweight compact nature of the actuator assembly makes it suitable for any application where mass and size are important considerations.\",\n \"Similar advantages are obtained by the use of the piezoelectric transducer to generate an electrical signal relative to magnet-moving coil systems.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473798"}}},{"rowIdx":754,"cells":{"text":{"kind":"list like","value":[["Installation for measuring pressure of at least one aeroplane wheel tyre","The invention concerns and installation (12) for measuring the pressure of at least one tyre of a wheel (16, 24) of an aeroplane (10) comprising: at least a pressure measuring sensor (26) associated with a tyre; a unit (30) operating on the pressure measurement provided in the aeroplane (10); and radio-frequency transmission means including for the or each sensor, a revolving antenna (28) borne by the wheel (16, 24) and connected to each measuring sensor (26) and to a fixed antenna (34; 134) connected by a conductive cable (32) to the unit operating (30) on the pressure measurement.","The fixed antenna (34; 134) is adapted to be borne by the aeroplane structure away from the or each wheel (16, 24) at least in an upper part of the shock strut (18, 20, 22) of the landing gear (14, 20) bearing the wheel."],["1.Installation (12) for measuring the pressure of at least one tyre of a wheel (16, 24) of an aeroplane (10), comprising: at least one pressure measuring sensor (26) associated with a tyre; a unit (30) operating on the pressure measurement, provided in the aeroplane (10); and radio-frequency transmission means comprising on the one hand, for the or each sensor, a revolving antenna (28) borne by the wheel (16, 24) and connected to each measuring sensor (26) and, on the other hand, a fixed antenna (34; 134) connected by a conductive cable (32) to the unit (30) operating on the pressure measurement, characterized in that the fixed antenna (34; 134) is adapted to be carried by the structure of the aeroplane at a spacing from the or each wheel (16, 24) at least in an upper part of the shock strut (18, 20; 122) of a landing gear element (14, 20) carrying the wheel.","2.Measuring installation according to claim 1, characterized in that the sock strut (18, 20; 122) has two movable sections (122A, 122B) connected by a shock absorber (122c), and the fixed antenna (134) is carried by the upper part (122a) of the shock strut (122) on the movable section (122A) which is integral with the aircraft fuselage.","3.Measuring installation according to claim 1 characterized in that the fixed antenna (34) is carried by the aircraft fuselage.","4.Installation according to claim 1, characterized in that said radio-frequency transmission means are adapted for the radio-frequencey transmission of a power signal for supplying the or each pressure measuring sensor (26).","5.Installation according to claim 1, characterized in that it comprises a plurality of sensors (26) associated with a plurality of movable antennas (28), and in that the transmission means comprise means for discriminating between the signals emitted from each movable antenna (28).","6.Installation according to claim 1, characterized in that the transmission frequency is between 100 kHz.","7.Aeroplane comprising an installation for measuring the pressure of at least one tyre of a wheel according to claim 1."],["The present invention relates to an installation for measuring the pressure of at least one tyre of an aeroplane wheel, of the type comprising: at least one pressure measuring sensor associated with a tyre; a unit for operating on the pressure measurement provided in the aeroplane; and radio-frequency transmission means comprising on the one hand, for the or each sensor, a revolving antenna borne by the wheel and connected to each measuring sensor and, on the other hand, a fixed antenna connected by a conductive cable to the unit for operating on the pressure measurement.","In aircraft it is useful for the cockpit to know the pressure of the tyres on the landing gear wheels.","To this end it is known to mount, on the rim of each wheel, a pressure sensor adapted to measure the inflation pressure of the tyre carried by the rim.","This pressure sensor sends the measurement to an operating unit provided in the body of the aircraft, and particularly in the cockpit.","This operating unit is adapted, for example, to provide the pilot with a display on a suitable screen showing the digital value of the pressure of each tyre.","To ensure that the data is transmitted from the sensor to the cockpit it is known to provide data transmission means between the fixed parts and moving parts of the landing gear.","In particular, it is known to provide, on the hub of the wheel and on the wheel axle about which the hub is mounted to rotate, two concentric coils together forming a transformer.","The coil borne by the hub is connected to the pressure sensor while the coil borne by the wheel axle is connected to the data processing unit.","The signal supplied by the sensor to the coil borne by the hub and corresponding to the level of pressure in the tyre induces a signal in the coil borne by the wheel axle of the landing gear.","This signal is analysed by the operating unit in order to derive from it the pressure in the tyre and make this available to the crew.","This solution works in a satisfactory manner.","However, the presence of two concentric coils makes the arrangement relatively bulky.","It has been envisaged to use antennas to provide a radio-frequency transmission between the moving part of the landing gear and the fixed part.","In the installations envisaged, an antenna borne by the fixed part of the landing gear is disposed on the hub of each wheel facing a corresponding rotary antenna borne by the moving part of the wheel.","The fixed antenna carried by the hub is connected to the unit for operating on the values, which is situated in the cockpit, by a data transmission cable.","Thus, the cable passes from the wheel hub along the length of the shock strut of the landing gear then into the fuselage of the aeroplane.","In large aircraft, the shock strut of each element of the landing gear comprises two successive sections connected to each other via a shock absorber.","With the two successive sections rendered movable by the presence of the shock absorber, the passage of the data transmission cable in the region of the shock absorber is tricky.","In particular, the cable is subjected to repeated mechanical stresses causing accidental breakage of the transmission cable in this region.","The invention sets out to propose an installation for measuring the tyre pressures of an aircraft which does not have the drawbacks described above and which is more reliable.","To this end, the invention relates to an installation for measuring the pressure of at least one tyre of an aeroplane wheel, of the type described above, characterised in that the fixed antenna is adapted to be borne by the structure of the aeroplane at a spacing from the or each wheel, at least in an upper part of the shock strut of an element of the landing gear carrying the wheel.","According to particular embodiments, the installation comprises one or more of the following features: the shock strut has two movable sections connected by a shock absorber, and the fixed antenna is carried by the upper part of the shock strut on the movable section which is integral with the aircraft fuselage; the fixed antenna is carried by the aircraft fuselage; said radio-frequency transmission means are adapted for the radio-frequency transmission of a power signal for supplying the or each pressure measuring sensor; it comprises a plurality of sensors associated with a plurality of movable antennas, and the transmission means comprise means for discriminating between the signals emitted from each movable antenna; and the transmission frequency is between 100 kHz and 150 kHz.","The invention also relates to an aeroplane comprising a pressure measuring installation as hereinbefore defined.","The invention will be better understood from reading the description that follows, which is provided solely by way of example and with reference to the drawings, wherein: FIG.","1 is a diagrammatic view of an aeroplane fitted with an installation for measuring the pressure of a tyre according to the invention; FIG.","2 is a diagrammatic view of the installation for measuring the pressure in two separate tyres of the same landing gear of an aeroplane; and FIG.","3 is a diagrammatic view of an element of the landing gear of an aeroplane on which is mounted an alternative embodiment of a pressure measuring installation according to the invention.","The aeroplane 10 shown in FIG.","1 is fitted with an installation 12 for measuring the pressure of each of the tyres of the aeroplane wheels.","As known per se, the aeroplane has at the front a landing gear element 14 comprising a number of wheels 16 arranged at the free end of a shock strut 18.Similarly the aeroplane comprises, underneath each wing, a landing gear element 20 comprising a shock strut 22 at the end of which are mounted a number of wheels 24.Each shock strut 18, 22 of the landing gear elements 14, 20 comprises two successive sections which are movable relative to one another and joined together by a shock absorber.","Each tyre fitted to a wheel 16, 24 of the aeroplane is associated with a pressure sensor generally designated 26.As known per se, the pressure sensor is borne for example by the rim of the wheel.","Each pressure sensor 26 is connected to a rotary antenna generally designated 28.This antenna 28 is fixedly attached to the rim and is mounted to rotate with the wheel relative to the shock strut of the landing gear element carrying the wheel.","Inside the aeroplane and especially in the cockpit there is a unit 30 for operating on the pressure measurement in each tyre of the aeroplane.","This operating unit 30 comprises, for example, a set of displays informing the pilot of the pressure level in each tyre.","This operating unit 30 is connected by a data transmission cable 32 to a fixed antenna 34 adapted to receive and transmit data from and to the antennas 28 associated with each sensor 26 mounted on the wheels of the aeroplane.","According to the invention, the fixed antenna 34 is carried by the structure of the aeroplane, at a spacing from each wheel, at least in the upper part of the shock strut of each landing gear element carrying a wheel.","More precisely, in the embodiment in FIG.","1, the fixed antenna 34 is fixed under the fuselage of the aircraft, in the front region thereof, close to the aircraft cockpit.","FIG.","2 shows the electronic structure of the radio-frequency data transmission means which permit communication between each of the pressure sensors and the operating unit 30.These transmission means are generally designated 38.In this Figure, only two sensors 26 and their associated movable antenna 28 are shown.","Each sensor 26 comprises a transducer 40 adapted to convert the pressure inside the tyre into an electric signal.","In addition, the sensor 26 comprises a data processing unit 42 comprising computing means 44 and coding and decoding means 46.The processing unit 42 is connected to a transmitting/receiving unit 48 to which the movable antenna 28 is connected.","The transmitting/receiving unit 48 comprises a demodulation stage 50 adapted to produce, from the radio-frequency signal received by the antenna 28, on the one hand a data signal 52 addressed to the coding and decoding means 46 and, on the other hand, a power signal 54 which supplies the data processing unit 42 on the one hand and all the stages of the transmitting/receiving unit 48.The electric power sent by the demodulation stage 50 comes from the radio-frequency signal received.","This radio-frequency signal is thus adapted for power transmission.","It has, for example, a frequency of between 100 kHz and 150 kHz.","This frequency is preferably 125 kHz.","The coding and decoding means 46 are adapted to interpret a pressure measurement command received from the cockpit through the radio-frequency communication.","In addition, the coding and decoding means 46 are adapted to code a pressure value produced by the processing means 44 for reading the transducer 40.This coding is carried out in particular in order to add to the pressure value, in the coded message, an identifier for the sensor 26, so that the pressure measurement value can be associated with a tyre.","Moreover, the transmitting/receiving unit 48 comprises a modulating stage 56 for the signal, which is adapted to convert the coded message coming from the coding and decoding means 46 into a signal suitable for radio-frequency transmission from the antenna 28.The modulation stage 56 is connected for this purpose to the antenna 28 in order to send the modulated signal to the fixed antenna 34.The antenna 34 is connected, like each antenna 28, to a transmitting/receiving unit 60.The latter is connected to a data processing unit 62, which is in turn connected to the unit operating on the values 30.This operating unit consists of a display, for example.","In addition, the transmitting/receiving unit 60, the data processing unit 62 and the operating unit 30 are all supplied by an electrical energy source 64.This supply is provided for example through the aeroplane's own power distribution network.","The transmitting/receiving unit 60 comprises a demodulation stage 66 which makes it possible to receive the modulated signal coming from the antenna 34 and to form from it a digital baseband signal which can be utilised by the data processing unit 62.Similarly, the communication unit 60 comprises a modulation stage 68 adapted to shape a signal coming from the data processing unit 62.In addition, this modulation stage 68 is adapted to combine, in a modulated signal, the data coming from the data processing unit 62 and a power signal for supplying the measuring sensors associated with the tyres.","The data processing unit 62 comprises coding and decoding means 70 for identifying, in the signal coming from the demodulating means 66, the origin of the pressure measurements as a function of the code provided by the pressure sensor which has sent the information.","Moreover, the coding and decoding means 70 effect the coding of the pressure requests sent to the modulating module 68 so that the request will be recognised by only one of the sensors.","The data processing unit 62 also comprises processing means 72 for shaping the pressure measurements received so that they can subsequently be processed by the operating means 30.In order to obtain a pressure measurement, the data processing unit 62 generates a request with a code appropriate to the sensor for the tyre whose pressure is to be recorded.","This coded signal is modulated by the modulating stage 68.Each of the sensors 26 receives the signal emitted by the antenna 34.The demodulation stage 50 of each sensor 26 carries out the demodulation of the signal, which is then sent to the data processing unit 42 and particularly to the coding and decoding means 46.If the request received contains a code which is recognised by the processing unit 42, the pressure measurement value returned by the transducer is shaped by the processing means 42 and encoded by the unit 46 before being modulated by the stage 56 for transmission.","After this, the signal thus modulated is sent from the antenna 28 to the antenna 34.If the sensor 26 does not recognise the request sent to it, it does not send any data back to the antenna 34.The signal received by the antenna 34 is demodulated by the demodulation stage 66, then sent to the coding and decoding means 70, which identify the sensor that has carried out the pressure measurement.","Thanks to the processing means 72, the pressure value is made available to the pilots of the aircraft by being displayed on the means 30 which operate on the pressure measurement value.","FIG.","3 shows an alternative embodiment of the installation according to the invention.","In this embodiment, the fixed antenna designated 134, which is analogous to the fixed antenna 34, is located on the upper part of the shock strut 122 of the landing gear element.","In particular, it is borne by the section 122A which is fixedly attached to the aircraft fuselage.","As before, each tyre is associated with a pressure sensor 26 connected to a movable antenna 28.In this embodiment, as in the previous one, a radio-frequency transmission is established between the rotating antenna 28 on each wheel and the fixed antenna 134.This radio-frequency transmission ensures both transmission of data and transmission of energy to supply the sensors.","The shock strut 122 comprises two successive sections 122A, 122B connected by a shock-absorbing element 122C.","In the embodiments in FIGS.","1 and 3, the movable antennas 28 and fixed antennas 34 or 134 are mounted at a spacing from one another, and particularly on either side of the shock absorbing element of the landing gear.","Thus, it will be seen that there are no wires running along the length of the shock absorber, thus reducing the risk of the cable breaking."]],"string":"[\n [\n \"Installation for measuring pressure of at least one aeroplane wheel tyre\",\n \"The invention concerns and installation (12) for measuring the pressure of at least one tyre of a wheel (16, 24) of an aeroplane (10) comprising: at least a pressure measuring sensor (26) associated with a tyre; a unit (30) operating on the pressure measurement provided in the aeroplane (10); and radio-frequency transmission means including for the or each sensor, a revolving antenna (28) borne by the wheel (16, 24) and connected to each measuring sensor (26) and to a fixed antenna (34; 134) connected by a conductive cable (32) to the unit operating (30) on the pressure measurement.\",\n \"The fixed antenna (34; 134) is adapted to be borne by the aeroplane structure away from the or each wheel (16, 24) at least in an upper part of the shock strut (18, 20, 22) of the landing gear (14, 20) bearing the wheel.\"\n ],\n [\n \"1.Installation (12) for measuring the pressure of at least one tyre of a wheel (16, 24) of an aeroplane (10), comprising: at least one pressure measuring sensor (26) associated with a tyre; a unit (30) operating on the pressure measurement, provided in the aeroplane (10); and radio-frequency transmission means comprising on the one hand, for the or each sensor, a revolving antenna (28) borne by the wheel (16, 24) and connected to each measuring sensor (26) and, on the other hand, a fixed antenna (34; 134) connected by a conductive cable (32) to the unit (30) operating on the pressure measurement, characterized in that the fixed antenna (34; 134) is adapted to be carried by the structure of the aeroplane at a spacing from the or each wheel (16, 24) at least in an upper part of the shock strut (18, 20; 122) of a landing gear element (14, 20) carrying the wheel.\",\n \"2.Measuring installation according to claim 1, characterized in that the sock strut (18, 20; 122) has two movable sections (122A, 122B) connected by a shock absorber (122c), and the fixed antenna (134) is carried by the upper part (122a) of the shock strut (122) on the movable section (122A) which is integral with the aircraft fuselage.\",\n \"3.Measuring installation according to claim 1 characterized in that the fixed antenna (34) is carried by the aircraft fuselage.\",\n \"4.Installation according to claim 1, characterized in that said radio-frequency transmission means are adapted for the radio-frequencey transmission of a power signal for supplying the or each pressure measuring sensor (26).\",\n \"5.Installation according to claim 1, characterized in that it comprises a plurality of sensors (26) associated with a plurality of movable antennas (28), and in that the transmission means comprise means for discriminating between the signals emitted from each movable antenna (28).\",\n \"6.Installation according to claim 1, characterized in that the transmission frequency is between 100 kHz.\",\n \"7.Aeroplane comprising an installation for measuring the pressure of at least one tyre of a wheel according to claim 1.\"\n ],\n [\n \"The present invention relates to an installation for measuring the pressure of at least one tyre of an aeroplane wheel, of the type comprising: at least one pressure measuring sensor associated with a tyre; a unit for operating on the pressure measurement provided in the aeroplane; and radio-frequency transmission means comprising on the one hand, for the or each sensor, a revolving antenna borne by the wheel and connected to each measuring sensor and, on the other hand, a fixed antenna connected by a conductive cable to the unit for operating on the pressure measurement.\",\n \"In aircraft it is useful for the cockpit to know the pressure of the tyres on the landing gear wheels.\",\n \"To this end it is known to mount, on the rim of each wheel, a pressure sensor adapted to measure the inflation pressure of the tyre carried by the rim.\",\n \"This pressure sensor sends the measurement to an operating unit provided in the body of the aircraft, and particularly in the cockpit.\",\n \"This operating unit is adapted, for example, to provide the pilot with a display on a suitable screen showing the digital value of the pressure of each tyre.\",\n \"To ensure that the data is transmitted from the sensor to the cockpit it is known to provide data transmission means between the fixed parts and moving parts of the landing gear.\",\n \"In particular, it is known to provide, on the hub of the wheel and on the wheel axle about which the hub is mounted to rotate, two concentric coils together forming a transformer.\",\n \"The coil borne by the hub is connected to the pressure sensor while the coil borne by the wheel axle is connected to the data processing unit.\",\n \"The signal supplied by the sensor to the coil borne by the hub and corresponding to the level of pressure in the tyre induces a signal in the coil borne by the wheel axle of the landing gear.\",\n \"This signal is analysed by the operating unit in order to derive from it the pressure in the tyre and make this available to the crew.\",\n \"This solution works in a satisfactory manner.\",\n \"However, the presence of two concentric coils makes the arrangement relatively bulky.\",\n \"It has been envisaged to use antennas to provide a radio-frequency transmission between the moving part of the landing gear and the fixed part.\",\n \"In the installations envisaged, an antenna borne by the fixed part of the landing gear is disposed on the hub of each wheel facing a corresponding rotary antenna borne by the moving part of the wheel.\",\n \"The fixed antenna carried by the hub is connected to the unit for operating on the values, which is situated in the cockpit, by a data transmission cable.\",\n \"Thus, the cable passes from the wheel hub along the length of the shock strut of the landing gear then into the fuselage of the aeroplane.\",\n \"In large aircraft, the shock strut of each element of the landing gear comprises two successive sections connected to each other via a shock absorber.\",\n \"With the two successive sections rendered movable by the presence of the shock absorber, the passage of the data transmission cable in the region of the shock absorber is tricky.\",\n \"In particular, the cable is subjected to repeated mechanical stresses causing accidental breakage of the transmission cable in this region.\",\n \"The invention sets out to propose an installation for measuring the tyre pressures of an aircraft which does not have the drawbacks described above and which is more reliable.\",\n \"To this end, the invention relates to an installation for measuring the pressure of at least one tyre of an aeroplane wheel, of the type described above, characterised in that the fixed antenna is adapted to be borne by the structure of the aeroplane at a spacing from the or each wheel, at least in an upper part of the shock strut of an element of the landing gear carrying the wheel.\",\n \"According to particular embodiments, the installation comprises one or more of the following features: the shock strut has two movable sections connected by a shock absorber, and the fixed antenna is carried by the upper part of the shock strut on the movable section which is integral with the aircraft fuselage; the fixed antenna is carried by the aircraft fuselage; said radio-frequency transmission means are adapted for the radio-frequency transmission of a power signal for supplying the or each pressure measuring sensor; it comprises a plurality of sensors associated with a plurality of movable antennas, and the transmission means comprise means for discriminating between the signals emitted from each movable antenna; and the transmission frequency is between 100 kHz and 150 kHz.\",\n \"The invention also relates to an aeroplane comprising a pressure measuring installation as hereinbefore defined.\",\n \"The invention will be better understood from reading the description that follows, which is provided solely by way of example and with reference to the drawings, wherein: FIG.\",\n \"1 is a diagrammatic view of an aeroplane fitted with an installation for measuring the pressure of a tyre according to the invention; FIG.\",\n \"2 is a diagrammatic view of the installation for measuring the pressure in two separate tyres of the same landing gear of an aeroplane; and FIG.\",\n \"3 is a diagrammatic view of an element of the landing gear of an aeroplane on which is mounted an alternative embodiment of a pressure measuring installation according to the invention.\",\n \"The aeroplane 10 shown in FIG.\",\n \"1 is fitted with an installation 12 for measuring the pressure of each of the tyres of the aeroplane wheels.\",\n \"As known per se, the aeroplane has at the front a landing gear element 14 comprising a number of wheels 16 arranged at the free end of a shock strut 18.Similarly the aeroplane comprises, underneath each wing, a landing gear element 20 comprising a shock strut 22 at the end of which are mounted a number of wheels 24.Each shock strut 18, 22 of the landing gear elements 14, 20 comprises two successive sections which are movable relative to one another and joined together by a shock absorber.\",\n \"Each tyre fitted to a wheel 16, 24 of the aeroplane is associated with a pressure sensor generally designated 26.As known per se, the pressure sensor is borne for example by the rim of the wheel.\",\n \"Each pressure sensor 26 is connected to a rotary antenna generally designated 28.This antenna 28 is fixedly attached to the rim and is mounted to rotate with the wheel relative to the shock strut of the landing gear element carrying the wheel.\",\n \"Inside the aeroplane and especially in the cockpit there is a unit 30 for operating on the pressure measurement in each tyre of the aeroplane.\",\n \"This operating unit 30 comprises, for example, a set of displays informing the pilot of the pressure level in each tyre.\",\n \"This operating unit 30 is connected by a data transmission cable 32 to a fixed antenna 34 adapted to receive and transmit data from and to the antennas 28 associated with each sensor 26 mounted on the wheels of the aeroplane.\",\n \"According to the invention, the fixed antenna 34 is carried by the structure of the aeroplane, at a spacing from each wheel, at least in the upper part of the shock strut of each landing gear element carrying a wheel.\",\n \"More precisely, in the embodiment in FIG.\",\n \"1, the fixed antenna 34 is fixed under the fuselage of the aircraft, in the front region thereof, close to the aircraft cockpit.\",\n \"FIG.\",\n \"2 shows the electronic structure of the radio-frequency data transmission means which permit communication between each of the pressure sensors and the operating unit 30.These transmission means are generally designated 38.In this Figure, only two sensors 26 and their associated movable antenna 28 are shown.\",\n \"Each sensor 26 comprises a transducer 40 adapted to convert the pressure inside the tyre into an electric signal.\",\n \"In addition, the sensor 26 comprises a data processing unit 42 comprising computing means 44 and coding and decoding means 46.The processing unit 42 is connected to a transmitting/receiving unit 48 to which the movable antenna 28 is connected.\",\n \"The transmitting/receiving unit 48 comprises a demodulation stage 50 adapted to produce, from the radio-frequency signal received by the antenna 28, on the one hand a data signal 52 addressed to the coding and decoding means 46 and, on the other hand, a power signal 54 which supplies the data processing unit 42 on the one hand and all the stages of the transmitting/receiving unit 48.The electric power sent by the demodulation stage 50 comes from the radio-frequency signal received.\",\n \"This radio-frequency signal is thus adapted for power transmission.\",\n \"It has, for example, a frequency of between 100 kHz and 150 kHz.\",\n \"This frequency is preferably 125 kHz.\",\n \"The coding and decoding means 46 are adapted to interpret a pressure measurement command received from the cockpit through the radio-frequency communication.\",\n \"In addition, the coding and decoding means 46 are adapted to code a pressure value produced by the processing means 44 for reading the transducer 40.This coding is carried out in particular in order to add to the pressure value, in the coded message, an identifier for the sensor 26, so that the pressure measurement value can be associated with a tyre.\",\n \"Moreover, the transmitting/receiving unit 48 comprises a modulating stage 56 for the signal, which is adapted to convert the coded message coming from the coding and decoding means 46 into a signal suitable for radio-frequency transmission from the antenna 28.The modulation stage 56 is connected for this purpose to the antenna 28 in order to send the modulated signal to the fixed antenna 34.The antenna 34 is connected, like each antenna 28, to a transmitting/receiving unit 60.The latter is connected to a data processing unit 62, which is in turn connected to the unit operating on the values 30.This operating unit consists of a display, for example.\",\n \"In addition, the transmitting/receiving unit 60, the data processing unit 62 and the operating unit 30 are all supplied by an electrical energy source 64.This supply is provided for example through the aeroplane's own power distribution network.\",\n \"The transmitting/receiving unit 60 comprises a demodulation stage 66 which makes it possible to receive the modulated signal coming from the antenna 34 and to form from it a digital baseband signal which can be utilised by the data processing unit 62.Similarly, the communication unit 60 comprises a modulation stage 68 adapted to shape a signal coming from the data processing unit 62.In addition, this modulation stage 68 is adapted to combine, in a modulated signal, the data coming from the data processing unit 62 and a power signal for supplying the measuring sensors associated with the tyres.\",\n \"The data processing unit 62 comprises coding and decoding means 70 for identifying, in the signal coming from the demodulating means 66, the origin of the pressure measurements as a function of the code provided by the pressure sensor which has sent the information.\",\n \"Moreover, the coding and decoding means 70 effect the coding of the pressure requests sent to the modulating module 68 so that the request will be recognised by only one of the sensors.\",\n \"The data processing unit 62 also comprises processing means 72 for shaping the pressure measurements received so that they can subsequently be processed by the operating means 30.In order to obtain a pressure measurement, the data processing unit 62 generates a request with a code appropriate to the sensor for the tyre whose pressure is to be recorded.\",\n \"This coded signal is modulated by the modulating stage 68.Each of the sensors 26 receives the signal emitted by the antenna 34.The demodulation stage 50 of each sensor 26 carries out the demodulation of the signal, which is then sent to the data processing unit 42 and particularly to the coding and decoding means 46.If the request received contains a code which is recognised by the processing unit 42, the pressure measurement value returned by the transducer is shaped by the processing means 42 and encoded by the unit 46 before being modulated by the stage 56 for transmission.\",\n \"After this, the signal thus modulated is sent from the antenna 28 to the antenna 34.If the sensor 26 does not recognise the request sent to it, it does not send any data back to the antenna 34.The signal received by the antenna 34 is demodulated by the demodulation stage 66, then sent to the coding and decoding means 70, which identify the sensor that has carried out the pressure measurement.\",\n \"Thanks to the processing means 72, the pressure value is made available to the pilots of the aircraft by being displayed on the means 30 which operate on the pressure measurement value.\",\n \"FIG.\",\n \"3 shows an alternative embodiment of the installation according to the invention.\",\n \"In this embodiment, the fixed antenna designated 134, which is analogous to the fixed antenna 34, is located on the upper part of the shock strut 122 of the landing gear element.\",\n \"In particular, it is borne by the section 122A which is fixedly attached to the aircraft fuselage.\",\n \"As before, each tyre is associated with a pressure sensor 26 connected to a movable antenna 28.In this embodiment, as in the previous one, a radio-frequency transmission is established between the rotating antenna 28 on each wheel and the fixed antenna 134.This radio-frequency transmission ensures both transmission of data and transmission of energy to supply the sensors.\",\n \"The shock strut 122 comprises two successive sections 122A, 122B connected by a shock-absorbing element 122C.\",\n \"In the embodiments in FIGS.\",\n \"1 and 3, the movable antennas 28 and fixed antennas 34 or 134 are mounted at a spacing from one another, and particularly on either side of the shock absorbing element of the landing gear.\",\n \"Thus, it will be seen that there are no wires running along the length of the shock absorber, thus reducing the risk of the cable breaking.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473808"}}},{"rowIdx":755,"cells":{"text":{"kind":"list like","value":[["Arc welding method","A method for controlling an arc welding equipment (21) used in a welding operation and adjustable by varying at least one welding parameter value, comprising the steps of determining said at least one welding parameter while using a theoretical model (24) of the welding process associated with the welding operation, and controlling the operation of the welding equipment and the welding process associated therewith by using said at least one welding parameter for adjusting the welding equipment and a device comprising an arc welding equipment (21) used in a welding operation and adjustable by varying at least one welding parameter and an arrangement (22) for controlling the operation of the arc welding equipment, said control arrangement including a means (23) adapted to determine the value of said at least one welding parameter while using a theoretical model (24) of the welding process, and a member (25) adapted to control the operation of the welding equipment and the welding process associated therewith by using said at least one welding parameter value for adjusting the welding equipment."],["1.A method for controlling an arc welding equipment used in a welding operation and adjustable by varying at least one welding parameter value, comprising: determining said at least one welding parameter value while using a theoretical model of the welding process associated with the welding operation, controlling the operation of the welding equipment and the welding process associated therewith by using said at least one welding parameter value for adjusting the welding equipment, dividing the welding process into at least two separate parts, of which at least one part represents the arc or a portion thereof, in the theoretical model, representing each of said at least two welding process parts and the welding parameter/parameters associated therewith by a model component, putting the model components and a model power source in an electric circuit model, and calculating at least one electric circuit model parameter value related to said at least one welding parameter from the electric circuit model.","2.A method for simulating an arc welding process, comprising: inputting data into a theoretical model representing the welding process, determining at least one welding parameter value of the welding process while using the theoretical model and the data input, with the purpose to simulate the welding process, dividing the welding process into at least two separate parts, of which at least one part represents the arc or a portion thereof, in the theoretical model, representing each of said at least two welding process parts and the welding parameter/parameters associated therewith by a model component, putting the model components and a model power source in an electric circuit model, and calculating at least one electric circuit model parameter value related to said at least one welding parameter from the electric circuit model.","3.A method for predicting the quality of a weld obtained from an arc welding operation, comprising: inputting data into a theoretical model representing the welding process associated with the welding operation, determining at least one property of the weld related to at least one welding parameter of the welding process while using the theoretical model and the data input, dividing the welding process into at least two separate parts, of which at least one part represents the arc or a portion thereof, in the theoretical model, representing each of said at least two welding process parts and the welding parameter/parameters associated therewith by a model component, putting the model components and a model power source in an electric circuit model, and calculating at least one electric circuit model parameter value related to said at least one welding parameter, and related to said at least one property of the weld, from the electric circuit model.","4.The method according to claim 1, further comprising: dividing the welding process so that the arc-wire interaction region part of the welding process is separately represented by one of said model components in the electric circuit model.","5.The method according to claim 1, further comprising: dividing the welding process so that the arc-work piece interaction region part of the welding process is separately represented by one of said model components in the electric circuit model.","6.The method according to claim 1, further comprising: dividing the welding process so that the arc-wire interaction region part of the welding process is separately represented by one of said model components in the electric circuit model, and/or dividing the welding process so that the arc-work piece interaction region part of the welding process is separately represented by one of said model components in the electric circuit model.","7.The method according to claim 1, further comprising: dividing the welding process so that the arc region part of the welding process is separately represented by one of said model components in the electric circuit model.","8.The method according to claim 7, further comprising: determining said model component representing the arc region part by means of information obtained through calculations from a physical model of the arc column region part of the welding process.","9.The method according to claim 7, further comprising: determining said model component representing the arc region part by means of information obtained through calculations from a physical model of the arc-wire interaction region part of the welding process.","10.The method according to claim 4, further comprising dividing the arc-wire interaction region part of the welding process into a first zone nearest to the wire and a second zone nearest to the arc column for describing the properties of the zones through different physical submodels.","11.The method according to claim 7, further comprising: determining said model component representing the arc region part by means of information obtained through calculations from a physical model of the arc-work piece interaction region part of the welding process.","12.The method according to claim 5, further comprising: dividing the arc-work piece interaction region part of the welding process into a first zone nearest to the work piece and a second zone nearest to the arc column for describing the properties of the zones through different physical submodels.","13.The method according to claim 7, further comprising: determining said model component representing the arc region part by means of said physical model describing the arc by at least one equation independent of the extension in space of the arc.","14.The method according to claim 13, further comprising: determining said model component representing the arc region part by means of said physical model describing the arc by said at least one equation only dependent on the time.","15.The method according to claim 1, further comprising: dividing the welding process so that the arc column region part of the welding process is separately represented by one of said model components in the electric circuit model.","16.The method according to claim 1, further comprising: dividing the welding process so that the wire part of the welding process is separately represented by one of said model components in the electric circuit model.","17.The method according to claim 1, further comprising: dividing the welding process so that the work piece part of the welding process is separately represented by one of said model components in the electric circuit model.","18.A device comprising an arc welding equipment used in a welding operation and adjustable by varying at least one welding parameter and an arrangement for controlling the operation of the arc welding equipment, said control arrangement including a means adapted to determine the value of said at least one welding parameter while using a theoretical model of the welding process, and a member adapted to control the operation of the welding equipment and the welding process associated therewith by using said at least one welding parameter value for adjusting the welding equipment, wherein the means is adapted to use said theoretical model comprising at least two separate parts, of which at least one part represents the arc or a portion thereof, corresponding to different parts of the welding process, each model part being represented by a model component, said model components together with a model power source being included in an electric circuit model and that said means is adapted to calculate at least one electric circuit model parameter value related to said at least one welding parameter from the electric circuit model for said control by said member.","19.The device according to claim 18, wherein the means is adapted to use said theoretical model having the arc-wire interaction region part of the welding process separately represented by one of said model components in the electric circuit model.","20.The device according to claim 17, wherein the means is adapted to use said theoretical model having the arc-work piece interaction region part of the welding process separately represented by one of said model components in the electric circuit model.","21.The device according to claim 18, wherein the means is adapted to use said theoretical model having the arc-wire interaction region part of the welding process separately represented by one of said model components in the electric circuit model and/or the arc-work piece interaction region part of the welding process separately represented by one of said model components in the electric circuit model.","22.The device according to claim 18, wherein the means is adapted to use said theoretical model having the arc region part of the welding process separately represented by one of said model components in the electric circuit model.","23.The device according to claim 22, wherein the means is adapted to use said theoretical model having information obtained through calculations from a physical model of the arc column region part of the welding process to be used in determining said model component representing the arc region part.","24.The device according to claim 22, wherein the means is adapted to use said theoretical model having information obtained through calculations from a physical model of the arc-wire interaction region part of the welding process to be used in determining said model component representing the arc region part.","25.The device according to claim 19, wherein the means is adapted to use said theoretical model having the arc-wire interaction region part of the welding process divided into a first zone nearest to the wire and a second zone nearest to the arc column for describing the properties of the zones through different physical submodels.","26.The device according to claim 22, wherein the means is adapted to use said theoretical model having information obtained through calculations from a physical model of the arc-work piece interaction region part of the welding process to be used in determining said model component representing the arc region part.","27.The device according to claim 20, wherein the means is adapted to use said theoretical model having the arc-work piece interaction region part of the welding process divided into a first zone nearest to the work piece and a second zone nearest to the arc column for describing the properties of the zones through different physical submodels.","28.The device according to claim 22, wherein the means is adapted to use said model component determined by means of said physical model describing the arc by at least one equation independent of the extension in space of the arc.","29.The device according to claim 28, wherein the means is adapted to use said model component determined by means of said physical model describing the arc by said at least one equation only dependent on the time.","30.The device according to claim 18, wherein the means is adapted to use said theoretical model having the arc column region part of the welding process separately represented by one of said model components in the electric circuit model.","31.The device according to claim 18, wherein the means is adapted to use said theoretical model having the wire part of the welding process separately represented by one of said model components in the electric circuit model.","32.The device according to claim 18, wherein the means is adapted to use said theoretical model having the work piece part of the welding process separately represented by one of said model components in the electric circuit model.","33.A use of a method according to claim 2, for predicting the metal droplet formation in an arc welding operation for a given set of welding parameters.","34.A use of a method according to claim 2, for predicting the metal transfer mode in an arc welding operation for a given set of welding parameters.","35.A use of a method according to claim 1, for calculating at least one welding parameter value required for obtaining a particular metal transfer mode in a welding operation.","36.A use of a method according to claim 1, for calculating at least one default setting value to be used for adjusting an automatic arc welding equipment prior to welding.","37.A computer program product for operating a method according to claim 1.38.A computer program product comprising code means and/or software code portions for enabling a processor to perform the steps of claim 1.39.A computer program product for operating a device according to claim 18.40.A computer program product according to claim 37, provided at least partially through a network such as the Internet.","41.A computer readable medium containing a computer program product according to claim 37."],["<SOH> FIELD OF THE INVENTION AND PRIOR ART <EOH>The present invention relates to a method for controlling an arc welding equipment used in a welding operation and adjustable by varying at least one welding parameter value, which method comprises the steps of: determining said at least one welding parameter value while using a theoretical model of the welding process associated with the welding operation and controlling the operation of the welding equipment and the welding process associated therewith by using said at least one welding parameter value for adjusting the welding equipment, to a method for simulating an arc welding process which method comprises the steps of: inputting data into a theoretical model representing the welding process and determining at least one welding parameter value of the welding process while using the theoretical model and the data input, with the purpose to simulate the welding process, and to a method for predicting the quality of a weld obtained from an arc welding operation which method comprises the steps of: inputting data into a theoretical model representing the welding process associated with the welding operation and determining at least one property of the weld related to at least one welding parameter of the welding process while using the theoretical model and the data input.","The invention also relates to a device comprising an arc welding equipment used in a welding operation and adjustable by varying at least one welding parameter and an arrangement for controlling the operation of the arc welding equipment, said control arrangement including a means adapted to determine the value of said at least one welding parameter while using a theoretical model of the welding process, and a member adapted to control the operation of the welding equipment and the welding process associated therewith by using said at least one welding parameter for adjusting the welding equipment.","Furthermore, the invention relates to uses of the methods and to computer program products according to claims enclosed.","When work pieces are joined by arc welding the quality of the welded joints is strongly dependent upon the parameters set during the arc welding operation.","Examples of these welding parameters are welding current, supply voltage, type of shielding gas, shielding gas flow rate, welding speed, torch angle, electrode (wire) extension, wire feed rate, and other properties of the wire and of the work piece, such as dimensions and materials.","These welding parameters interact with each other in the welding process in a complex manner and influence the properties, i.e.","the geometry or type of weld, metallurgy and the mechanical strength of the weld resulting from the welding operation.","More precisely, the quality of a weld joint depends strongly on the metal transfer mode present during the welding.","In other words, the way metal is transferred from the wire tip (for example the anode) to the work piece (for example the cathode) during the welding has a very strong effect on the final properties and the quality of the weld joint.","There is of course a need to know how different sets of the adjustable parameters will effect the properties of the weld joint, so that the parameters may be adjusted in a way resulting in the weld desired.","One way is to attempt to predict the metal transfer mode based on the knowledge about the influence of the different parameters.","However, according to the state of art the metal transfer mode is mainly determined through empirical methods.","The metal transfer mode may be identified through the noise generated by the welding process by people with a very long experience in arc welding.","Inspection of the weld joint after finishing the welding operation may also give clues to which metal transfer mode that was present during the welding operation.","Thus, at present, the settings of important welding process parameters used in robotic arc welding are found out experimentally by trial and error for every single welding task or welding condition, which is both time consuming and costly.","For example, in case of using parameters resulting in an undesired metal transfer mode the welded joint may be of a poor quality and the work piece may be destroyed.","One reason to the lack of knowledge about how the welding parameters effect the welding process in general, and particularly the arc, and about the relations between the welding parameters used and the quality of the weld joint, is the difficulty in making measurements of the arc, or in the vicinity of the arc, and in the near to arc regions close to the wire and the work piece.","Thus, as an undesired consequence it is very difficult to design a control system based upon direct measurements of the arc parameters for controlling an arc welding equipment.","The fact is that automated or robotic arc welding are used in quite few of the arc welding applications performed throughout the world today, since the difficulty in predicting the result of the welding operation and the properties of welding process associated thereto for a given set of welding parameters, together with proportionately small series of work pieces, makes it more cost efficient to perform the work manually by workers skilled in welding."],["<SOH> SUMMARY OF THE INVENTION <EOH>A first object of the invention is to provide a method for controlling an arc welding equipment, which method enabling that the arc welding equipment may be controlled during a welding operation by adjusting at least one welding parameter determined without the need of measurements of the welding process or repeated welding experiments prior to welding.","The first object is obtained according to the invention by providing such a method, referred to in the introduction, which comprises the steps of: dividing the welding process into at least two separate parts, of which at least one part represents the arc or a portion thereof, in the theoretical model, representing each of said at least two welding process parts and the welding parameter/parameters associated therewith by a model component, putting the model components and a model power source in an electric circuit model, and calculating at least one electric circuit model parameter related to said at least one welding parameter from the electric circuit model.","By such a method it is possible to determine at least one welding parameter value, such as the welding current or voltage supply, wire feed rate, wire extension etc., and to use this welding parameter value for controlling the arc welding equipment in accordance with the present conditions to obtain a weld with the properties desired.","A second object of the invention is to provide a method for simulating an arc welding process without the need of measurements of the corresponding real welding process or repeated welding experiments.","The second object is obtained according to the invention by providing such a method, referred to in the introduction, which comprises the steps of: dividing the welding process into at least two separate parts, of which at least one part represents the arc or a portion thereof, in the theoretical model, representing each of said at least two welding process parts and the welding parameter/parameters associated therewith by a model component, putting the model components and a model power source in an electric circuit model, and calculating at least one electric circuit model parameter related to said at least one welding parameter from the electric circuit model.","By this simulation method it is possible to obtain at least one welding parameter value, and/or the present metal transfer mode, for a given set of other welding parameters in the simulation, and for example this simulation method may be used for understanding and designing of real welding processes and for default setting of welding parameters of arc welding equipment in different arc welding operations.","A third object of the invention is to provide a method for predicting the quality of a weld obtained from an arc welding operation without the need of repeated welding experiments.","The third object is obtained according to the invention by providing such a method, referred to in the introduction, which comprises the steps of: dividing the welding process into at least two separate parts, of which at least one part represents the arc or a portion thereof, in the theoretical model, representing each of said at least two welding process parts and the welding parameter/parameters associated therewith by a model component, putting the model components and a model power source in an electric circuit model, and calculating at least one electric circuit model parameter related to said at least one welding parameter, and related to said at least one property of the weld, from the electric circuit model.","By such a method it is possible to predict one or more mechanical properties of a weld resulting from an arc welding operation by determining one or more welding parameters required for the quality desired and use this information in real arc welding operations.","It is possible to use the method for predicting the quality of a weld for a given set of welding parameters as well as for determining one or more welding parameters required for obtaining certain mechanical properties of the weld.","According to a preferred embodiment of the invention one or more of said methods comprise the step of: dividing the welding process so that the arc-wire interaction region part of the welding process is separately represented by one of said model components in the electric circuit model.","By such a method, together with a suitable physical model, important information is obtained because it is possible to take important factors in the welding process, such as the voltage drop over the region near to anode and the heat flux from arc to anode, into consideration in determining said at least one welding parameter value.","In the arc-wire interaction region of the welding process, high gradients of different quantities such as temperature and concentration of particles are present, and the voltage drop in this region is very high.","Thus, this region gives an important contribution to the characteristics of the welding process.","According to another preferred embodiment of the invention one or more of said methods comprise the step of: dividing the welding process so that the arc-work piece interaction region part of the welding process is separately represented by one of said model components in the electric circuit model.","By such a method, together with a suitable physical model, important information is obtained because it is possible to take important factors in the welding process, such as the voltage drop over the region near to cathode and the heat flux from arc to cathode, into consideration in determining said at least one welding parameter value.","In the arc-work piece interaction region of the welding process, high gradients of different quantities such as temperature and concentration of particles are present, and the voltage drop in this region is very high.","Thus, this region gives an important contribution to the characteristics of the welding process.","According to another preferred embodiment of the invention one or more of said methods comprise the step of: dividing the welding process so that the arc region part of the welding process is separately represented by one of said model components in the electric circuit model.","By such a method, and particularly together with an embodiment comprising the step of: determining said model component representing the arc region part by means of information obtained through calculations from a physical model of the arc column region part of the welding process, important information is obtained such as arc mean temperature, temperature near to the wire, temperature near to the work piece, arc radius and arc current-voltage characteristics, and it is also possible to take the thermophysical properties of a shielding gas present in the arc region into consideration in determining said at least one welding parameter value.","According to another preferred embodiment of the invention one or more of said methods comprise the step of: determining said model component representing the arc region part by means of information obtained through calculations from a physical model of the arc-wire interaction region part and/or of the arc-work piece interaction region part of the welding process, important information is obtained because it is possible to take important factors in the welding process, such as the voltage drop over the region near to anode/cathode and the heat flux from arc to anode/cathode, into consideration in determining said at least one welding parameter value.","In the arc-wire interaction region and in the arc-work piece interaction region of the welding process, high gradients of different quantities such as temperature and concentration of particles are present, and the voltage drop in these regions is very high.","Thus, these regions give an important contribution to the characteristics of the welding process.","According to another preferred embodiment of the invention one or more of said methods comprise the step of: determining said model component representing the arc region part by means of said physical model describing the arc by at least one equation independent of the extension in space of the arc.","By such a method, and particularly together with an embodiment comprising the step of: determining said model component representing the arc region part by means of said physical model describing the arc by said at least one equation only dependent on the time, the model component and thus said at least one welding parameter value, may be determined fast and with proportionally short computational time, which makes the method suitable to be used for example in controlling an arc welding equipment on line.","According to another preferred embodiment of the invention one or more of said methods comprise the step of: dividing the welding process so that the arc column region part of the welding process is separately represented by one of said model components in the electric circuit model.","By such a method, together with a suitable physical model, important information is obtained such as arc mean temperature, temperature near to the wire, temperature near to the work piece, arc radius and arc current-voltage characteristics, and it is also possible to take the thermophysical properties of a shielding gas present in the arc column region into consideration in determining said at least one welding parameter.","According to another preferred embodiment of the invention said methods comprise the step of: dividing the welding process so that the wire part of the welding process is separately represented by one of said model components in the electric circuit model.","By such a method, together with a suitable physical model, important information is obtained such as metal transfer mode, dipping frequency etc, and it is also possible to take material properties of the wire into consideration in determining the value of said at least one welding parameter.","According to another preferred embodiment of the invention said methods comprise the step of: dividing the welding process so that the work piece part of the welding process is separately represented by one of said model components in the electric circuit model.","By such a method, together with a suitable physical model, important information is obtained about quality features such as weld joint profile, penetration profile etc, and it is also possible to take material properties of the work piece into consideration in determining the value of said at least one welding parameter.","A fourth object of the invention is to provide a device which has an arrangement for controlling the arc welding equipment during a welding operation by adjusting at least one welding parameter determined without the need of measurements of the welding process or repeated welding experiments prior to welding.","The fourth object is obtained according to the invention by providing such a device, referred to in the introduction, in which the means included in the control arrangement is adapted to use said theoretical model comprising at least two separate parts, of which at least one part represents the arc or a portion thereof, corresponding to different parts of the welding process, each model part being represented by a model component, said model components together with a model power source being included in an electric circuit model, and adapted to calculate at least one electric circuit model parameter related to said at least one welding parameter from the electric circuit model for said control by said member.","Thus, by the means according to the invention it is possible to determine how to set at least one welding parameter value, such as the welding current or supply voltage, wire feed rate, wire extension etc., with the purpose of optimizing the welding process under the present conditions, and use this welding parameter value by said member for adjusting the arc welding equipment in accordance with the present conditions to obtain a weld with the properties desired.","According to a preferred embodiment of the invention said means is adapted to use said theoretical model having the arc-wire interaction region part of the welding process separately represented by one of said model components in the electric circuit model.","By such a device, together with a suitable physical model, important information is obtained because it is possible to take important factors in the welding process, such as the voltage drop over the region near to anode and the heat flux from arc to anode, into consideration in determining said at least one welding parameter value.","In the arc-wire interaction region of the welding process, high gradients of different quantities such as temperature and concentration of particles are present, and the voltage drop in this region is very high.","Thus, this region gives an important contribution to the characteristics of the welding process.","According to another preferred embodiment of the invention said means is adapted to use said theoretical model having the arc-work piece interaction region part of the welding process separately represented by one of said model components in the electric circuit model.","By such a device, together with a suitable physical model, important information is obtained because it is possible to take important factors in the welding process, such as the voltage drop over the region near to cathode and the heat flux from arc to cathode, into consideration in determining said at least one welding parameter value.","In the arc-work piece interaction region of the welding process, high gradients of different quantities such as temperature and concentration of particles are present, and the voltage drop in this region is very high.","Thus, this region gives an important contribution to the characteristics of the welding process.","According to another preferred embodiment of the invention said means is adapted to use said theoretical model having the arc region part of the welding process separately represented by one of said model components in the electric circuit model.","By such a device, and particularly together with an embodiment of the invention in which the means is adapted to use said theoretical model having information obtained through calculations from a physical model of the arc column region part of the welding process to be used in determining said model component representing the arc region part, important information is obtained such as arc mean temperature, temperature near to wire, temperature near to work piece, arc radius and arc current-voltage characteristics, and it is also possible to take the thermo-physical properties of a shielding gas present in the arc region into consideration in determining said at least one welding parameter.","According to another preferred embodiment of the invention said means is adapted to use said theoretical model having information obtained through calculations from a physical model of the arc-wire interaction region part and/or of the arc-work piece interaction region part of the welding process to be used in determining said model component representing the arc region part, important information is obtained because it is possible to take important factors in the welding process, such as the voltage drop over the region near to anode/cathode and the heat flux from arc to anode/cathode, into consideration in determining said at least one welding parameter value.","In the arc-wire interaction region and in the arc-work piece interaction region of the welding process, high gradients of different quantities such as temperature and concentration of particles are present, and the voltage drop in these regions is very high.","Thus, these regions give an important contribution to the characteristics of the welding process.","According to another preferred embodiment of the invention in which said means is adapted to use said model component determined by means of said physical model describing the arc by at least one equation independent of the extension in space of the arc, and particularly together with an embodiment of the invention in which said means is adapted to use said model component determined by means of said physical model describing the arc by said at least one equation only dependent on the time, the model component and thus said at least one welding parameter value, may be determined fast and with proportionally short computational time, which makes the device very suitable to be used in controlling the arc welding equipment on line.","According to another preferred embodiment of the invention said means is adapted to use said theoretical model having the arc column region part of the welding process separately represented by one of said model components in the electric circuit model.","By such a device, together with a suitable physical model, important information is obtained such as arc mean temperature, temperature near to wire, temperature near to work piece, arc radius and arc current-voltage characteristics, and it is also possible to take the thermo-physical properties of a shielding gas present in the arc column region into consideration in determining said at least one welding parameter.","According to another preferred embodiment of the invention said means is adapted to use said theoretical model having the wire part of the welding process separately represented by one of said model components.","By such a device, together with a suitable physical model, important information is obtained such as metal transfer mode, dipping frequency etc, and it is also possible to take material properties of the wire into consideration in determining the value of said at least one welding parameter.","According to another preferred embodiment of the invention said means is adapted to use said theoretical model having the work piece part of the welding process separately represented by one of said model components.","By such a device, together with a suitable physical model, important information is obtained about quality features such as weld joint profile, penetration profile etc, and it is also possible to take material properties of the work piece into consideration in determining the value of said at least one welding parameter.","The invention also relates to computer program products according to the claims enclosed.","It is obvious that the methods according to the invention defined in the sets of method claims enclosed are well suited to be performed through program instructions from a processor which may be effected by a computer program.","Further advantages of and features of the methods and the device, the computer program products, and the uses of the methods appear from the following detailed description and the other claims enclosed herein."],["FIELD OF THE INVENTION AND PRIOR ART The present invention relates to a method for controlling an arc welding equipment used in a welding operation and adjustable by varying at least one welding parameter value, which method comprises the steps of: determining said at least one welding parameter value while using a theoretical model of the welding process associated with the welding operation and controlling the operation of the welding equipment and the welding process associated therewith by using said at least one welding parameter value for adjusting the welding equipment, to a method for simulating an arc welding process which method comprises the steps of: inputting data into a theoretical model representing the welding process and determining at least one welding parameter value of the welding process while using the theoretical model and the data input, with the purpose to simulate the welding process, and to a method for predicting the quality of a weld obtained from an arc welding operation which method comprises the steps of: inputting data into a theoretical model representing the welding process associated with the welding operation and determining at least one property of the weld related to at least one welding parameter of the welding process while using the theoretical model and the data input.","The invention also relates to a device comprising an arc welding equipment used in a welding operation and adjustable by varying at least one welding parameter and an arrangement for controlling the operation of the arc welding equipment, said control arrangement including a means adapted to determine the value of said at least one welding parameter while using a theoretical model of the welding process, and a member adapted to control the operation of the welding equipment and the welding process associated therewith by using said at least one welding parameter for adjusting the welding equipment.","Furthermore, the invention relates to uses of the methods and to computer program products according to claims enclosed.","When work pieces are joined by arc welding the quality of the welded joints is strongly dependent upon the parameters set during the arc welding operation.","Examples of these welding parameters are welding current, supply voltage, type of shielding gas, shielding gas flow rate, welding speed, torch angle, electrode (wire) extension, wire feed rate, and other properties of the wire and of the work piece, such as dimensions and materials.","These welding parameters interact with each other in the welding process in a complex manner and influence the properties, i.e.","the geometry or type of weld, metallurgy and the mechanical strength of the weld resulting from the welding operation.","More precisely, the quality of a weld joint depends strongly on the metal transfer mode present during the welding.","In other words, the way metal is transferred from the wire tip (for example the anode) to the work piece (for example the cathode) during the welding has a very strong effect on the final properties and the quality of the weld joint.","There is of course a need to know how different sets of the adjustable parameters will effect the properties of the weld joint, so that the parameters may be adjusted in a way resulting in the weld desired.","One way is to attempt to predict the metal transfer mode based on the knowledge about the influence of the different parameters.","However, according to the state of art the metal transfer mode is mainly determined through empirical methods.","The metal transfer mode may be identified through the noise generated by the welding process by people with a very long experience in arc welding.","Inspection of the weld joint after finishing the welding operation may also give clues to which metal transfer mode that was present during the welding operation.","Thus, at present, the settings of important welding process parameters used in robotic arc welding are found out experimentally by trial and error for every single welding task or welding condition, which is both time consuming and costly.","For example, in case of using parameters resulting in an undesired metal transfer mode the welded joint may be of a poor quality and the work piece may be destroyed.","One reason to the lack of knowledge about how the welding parameters effect the welding process in general, and particularly the arc, and about the relations between the welding parameters used and the quality of the weld joint, is the difficulty in making measurements of the arc, or in the vicinity of the arc, and in the near to arc regions close to the wire and the work piece.","Thus, as an undesired consequence it is very difficult to design a control system based upon direct measurements of the arc parameters for controlling an arc welding equipment.","The fact is that automated or robotic arc welding are used in quite few of the arc welding applications performed throughout the world today, since the difficulty in predicting the result of the welding operation and the properties of welding process associated thereto for a given set of welding parameters, together with proportionately small series of work pieces, makes it more cost efficient to perform the work manually by workers skilled in welding.","SUMMARY OF THE INVENTION A first object of the invention is to provide a method for controlling an arc welding equipment, which method enabling that the arc welding equipment may be controlled during a welding operation by adjusting at least one welding parameter determined without the need of measurements of the welding process or repeated welding experiments prior to welding.","The first object is obtained according to the invention by providing such a method, referred to in the introduction, which comprises the steps of: dividing the welding process into at least two separate parts, of which at least one part represents the arc or a portion thereof, in the theoretical model, representing each of said at least two welding process parts and the welding parameter/parameters associated therewith by a model component, putting the model components and a model power source in an electric circuit model, and calculating at least one electric circuit model parameter related to said at least one welding parameter from the electric circuit model.","By such a method it is possible to determine at least one welding parameter value, such as the welding current or voltage supply, wire feed rate, wire extension etc., and to use this welding parameter value for controlling the arc welding equipment in accordance with the present conditions to obtain a weld with the properties desired.","A second object of the invention is to provide a method for simulating an arc welding process without the need of measurements of the corresponding real welding process or repeated welding experiments.","The second object is obtained according to the invention by providing such a method, referred to in the introduction, which comprises the steps of: dividing the welding process into at least two separate parts, of which at least one part represents the arc or a portion thereof, in the theoretical model, representing each of said at least two welding process parts and the welding parameter/parameters associated therewith by a model component, putting the model components and a model power source in an electric circuit model, and calculating at least one electric circuit model parameter related to said at least one welding parameter from the electric circuit model.","By this simulation method it is possible to obtain at least one welding parameter value, and/or the present metal transfer mode, for a given set of other welding parameters in the simulation, and for example this simulation method may be used for understanding and designing of real welding processes and for default setting of welding parameters of arc welding equipment in different arc welding operations.","A third object of the invention is to provide a method for predicting the quality of a weld obtained from an arc welding operation without the need of repeated welding experiments.","The third object is obtained according to the invention by providing such a method, referred to in the introduction, which comprises the steps of: dividing the welding process into at least two separate parts, of which at least one part represents the arc or a portion thereof, in the theoretical model, representing each of said at least two welding process parts and the welding parameter/parameters associated therewith by a model component, putting the model components and a model power source in an electric circuit model, and calculating at least one electric circuit model parameter related to said at least one welding parameter, and related to said at least one property of the weld, from the electric circuit model.","By such a method it is possible to predict one or more mechanical properties of a weld resulting from an arc welding operation by determining one or more welding parameters required for the quality desired and use this information in real arc welding operations.","It is possible to use the method for predicting the quality of a weld for a given set of welding parameters as well as for determining one or more welding parameters required for obtaining certain mechanical properties of the weld.","According to a preferred embodiment of the invention one or more of said methods comprise the step of: dividing the welding process so that the arc-wire interaction region part of the welding process is separately represented by one of said model components in the electric circuit model.","By such a method, together with a suitable physical model, important information is obtained because it is possible to take important factors in the welding process, such as the voltage drop over the region near to anode and the heat flux from arc to anode, into consideration in determining said at least one welding parameter value.","In the arc-wire interaction region of the welding process, high gradients of different quantities such as temperature and concentration of particles are present, and the voltage drop in this region is very high.","Thus, this region gives an important contribution to the characteristics of the welding process.","According to another preferred embodiment of the invention one or more of said methods comprise the step of: dividing the welding process so that the arc-work piece interaction region part of the welding process is separately represented by one of said model components in the electric circuit model.","By such a method, together with a suitable physical model, important information is obtained because it is possible to take important factors in the welding process, such as the voltage drop over the region near to cathode and the heat flux from arc to cathode, into consideration in determining said at least one welding parameter value.","In the arc-work piece interaction region of the welding process, high gradients of different quantities such as temperature and concentration of particles are present, and the voltage drop in this region is very high.","Thus, this region gives an important contribution to the characteristics of the welding process.","According to another preferred embodiment of the invention one or more of said methods comprise the step of: dividing the welding process so that the arc region part of the welding process is separately represented by one of said model components in the electric circuit model.","By such a method, and particularly together with an embodiment comprising the step of: determining said model component representing the arc region part by means of information obtained through calculations from a physical model of the arc column region part of the welding process, important information is obtained such as arc mean temperature, temperature near to the wire, temperature near to the work piece, arc radius and arc current-voltage characteristics, and it is also possible to take the thermophysical properties of a shielding gas present in the arc region into consideration in determining said at least one welding parameter value.","According to another preferred embodiment of the invention one or more of said methods comprise the step of: determining said model component representing the arc region part by means of information obtained through calculations from a physical model of the arc-wire interaction region part and/or of the arc-work piece interaction region part of the welding process, important information is obtained because it is possible to take important factors in the welding process, such as the voltage drop over the region near to anode/cathode and the heat flux from arc to anode/cathode, into consideration in determining said at least one welding parameter value.","In the arc-wire interaction region and in the arc-work piece interaction region of the welding process, high gradients of different quantities such as temperature and concentration of particles are present, and the voltage drop in these regions is very high.","Thus, these regions give an important contribution to the characteristics of the welding process.","According to another preferred embodiment of the invention one or more of said methods comprise the step of: determining said model component representing the arc region part by means of said physical model describing the arc by at least one equation independent of the extension in space of the arc.","By such a method, and particularly together with an embodiment comprising the step of: determining said model component representing the arc region part by means of said physical model describing the arc by said at least one equation only dependent on the time, the model component and thus said at least one welding parameter value, may be determined fast and with proportionally short computational time, which makes the method suitable to be used for example in controlling an arc welding equipment on line.","According to another preferred embodiment of the invention one or more of said methods comprise the step of: dividing the welding process so that the arc column region part of the welding process is separately represented by one of said model components in the electric circuit model.","By such a method, together with a suitable physical model, important information is obtained such as arc mean temperature, temperature near to the wire, temperature near to the work piece, arc radius and arc current-voltage characteristics, and it is also possible to take the thermophysical properties of a shielding gas present in the arc column region into consideration in determining said at least one welding parameter.","According to another preferred embodiment of the invention said methods comprise the step of: dividing the welding process so that the wire part of the welding process is separately represented by one of said model components in the electric circuit model.","By such a method, together with a suitable physical model, important information is obtained such as metal transfer mode, dipping frequency etc, and it is also possible to take material properties of the wire into consideration in determining the value of said at least one welding parameter.","According to another preferred embodiment of the invention said methods comprise the step of: dividing the welding process so that the work piece part of the welding process is separately represented by one of said model components in the electric circuit model.","By such a method, together with a suitable physical model, important information is obtained about quality features such as weld joint profile, penetration profile etc, and it is also possible to take material properties of the work piece into consideration in determining the value of said at least one welding parameter.","A fourth object of the invention is to provide a device which has an arrangement for controlling the arc welding equipment during a welding operation by adjusting at least one welding parameter determined without the need of measurements of the welding process or repeated welding experiments prior to welding.","The fourth object is obtained according to the invention by providing such a device, referred to in the introduction, in which the means included in the control arrangement is adapted to use said theoretical model comprising at least two separate parts, of which at least one part represents the arc or a portion thereof, corresponding to different parts of the welding process, each model part being represented by a model component, said model components together with a model power source being included in an electric circuit model, and adapted to calculate at least one electric circuit model parameter related to said at least one welding parameter from the electric circuit model for said control by said member.","Thus, by the means according to the invention it is possible to determine how to set at least one welding parameter value, such as the welding current or supply voltage, wire feed rate, wire extension etc., with the purpose of optimizing the welding process under the present conditions, and use this welding parameter value by said member for adjusting the arc welding equipment in accordance with the present conditions to obtain a weld with the properties desired.","According to a preferred embodiment of the invention said means is adapted to use said theoretical model having the arc-wire interaction region part of the welding process separately represented by one of said model components in the electric circuit model.","By such a device, together with a suitable physical model, important information is obtained because it is possible to take important factors in the welding process, such as the voltage drop over the region near to anode and the heat flux from arc to anode, into consideration in determining said at least one welding parameter value.","In the arc-wire interaction region of the welding process, high gradients of different quantities such as temperature and concentration of particles are present, and the voltage drop in this region is very high.","Thus, this region gives an important contribution to the characteristics of the welding process.","According to another preferred embodiment of the invention said means is adapted to use said theoretical model having the arc-work piece interaction region part of the welding process separately represented by one of said model components in the electric circuit model.","By such a device, together with a suitable physical model, important information is obtained because it is possible to take important factors in the welding process, such as the voltage drop over the region near to cathode and the heat flux from arc to cathode, into consideration in determining said at least one welding parameter value.","In the arc-work piece interaction region of the welding process, high gradients of different quantities such as temperature and concentration of particles are present, and the voltage drop in this region is very high.","Thus, this region gives an important contribution to the characteristics of the welding process.","According to another preferred embodiment of the invention said means is adapted to use said theoretical model having the arc region part of the welding process separately represented by one of said model components in the electric circuit model.","By such a device, and particularly together with an embodiment of the invention in which the means is adapted to use said theoretical model having information obtained through calculations from a physical model of the arc column region part of the welding process to be used in determining said model component representing the arc region part, important information is obtained such as arc mean temperature, temperature near to wire, temperature near to work piece, arc radius and arc current-voltage characteristics, and it is also possible to take the thermo-physical properties of a shielding gas present in the arc region into consideration in determining said at least one welding parameter.","According to another preferred embodiment of the invention said means is adapted to use said theoretical model having information obtained through calculations from a physical model of the arc-wire interaction region part and/or of the arc-work piece interaction region part of the welding process to be used in determining said model component representing the arc region part, important information is obtained because it is possible to take important factors in the welding process, such as the voltage drop over the region near to anode/cathode and the heat flux from arc to anode/cathode, into consideration in determining said at least one welding parameter value.","In the arc-wire interaction region and in the arc-work piece interaction region of the welding process, high gradients of different quantities such as temperature and concentration of particles are present, and the voltage drop in these regions is very high.","Thus, these regions give an important contribution to the characteristics of the welding process.","According to another preferred embodiment of the invention in which said means is adapted to use said model component determined by means of said physical model describing the arc by at least one equation independent of the extension in space of the arc, and particularly together with an embodiment of the invention in which said means is adapted to use said model component determined by means of said physical model describing the arc by said at least one equation only dependent on the time, the model component and thus said at least one welding parameter value, may be determined fast and with proportionally short computational time, which makes the device very suitable to be used in controlling the arc welding equipment on line.","According to another preferred embodiment of the invention said means is adapted to use said theoretical model having the arc column region part of the welding process separately represented by one of said model components in the electric circuit model.","By such a device, together with a suitable physical model, important information is obtained such as arc mean temperature, temperature near to wire, temperature near to work piece, arc radius and arc current-voltage characteristics, and it is also possible to take the thermo-physical properties of a shielding gas present in the arc column region into consideration in determining said at least one welding parameter.","According to another preferred embodiment of the invention said means is adapted to use said theoretical model having the wire part of the welding process separately represented by one of said model components.","By such a device, together with a suitable physical model, important information is obtained such as metal transfer mode, dipping frequency etc, and it is also possible to take material properties of the wire into consideration in determining the value of said at least one welding parameter.","According to another preferred embodiment of the invention said means is adapted to use said theoretical model having the work piece part of the welding process separately represented by one of said model components.","By such a device, together with a suitable physical model, important information is obtained about quality features such as weld joint profile, penetration profile etc, and it is also possible to take material properties of the work piece into consideration in determining the value of said at least one welding parameter.","The invention also relates to computer program products according to the claims enclosed.","It is obvious that the methods according to the invention defined in the sets of method claims enclosed are well suited to be performed through program instructions from a processor which may be effected by a computer program.","Further advantages of and features of the methods and the device, the computer program products, and the uses of the methods appear from the following detailed description and the other claims enclosed herein.","BRIEF DESCRIPTION OF THE DRAWINGS With reference to the appended drawings, below follows a description of preferred embodiments of the invention cited as examples.","In the drawings: FIG.","1 is a schematic illustration of an arc welding process, FIG.","2 is a schematic illustration of the voltage distribution in the gap between a wire and a work piece in an arc welding operation, FIG.","3 is an illustration of an electric circuit model representing an arc welding process, FIG.","4 is a schematic illustration of a device according to the invention and of applications of the invention, FIG.","5 is a block diagram of a method according to the invention, FIG.","6 is a block diagram of another method according to the invention, FIG.","7 is a block diagram of another method according to the invention, FIG.","8 is a schematic illustration of a physical model of the arc, FIG.","9 is an illustration of the geometry of an arc defined in a physical model, FIG.","10 is an illustration of a physical model of the arc-wire interaction region in an arc welding process, FIG.","11 is an illustration of a physical model of the arc-work piece interaction region in an arc welding process, FIG.","12 is an illustration of the energy balance in the ionization zone, FIG.","13 shows different kinds of welding parameters, FIG.","14 is a schematic definition for work piece modelling of a horizontal-vertical fillet joint produced by gas metal arc welding, FIG.","15 shows an example of a non-linear regression model, and FIGS.","16 and 17 are analytic models of work piece.","DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS OF THE INVENTION In this application the wording “welding process” is used as a generic term of the main parts included in an arc welding operation and of the physical processes associated therewith which take place when an electric arc is generated between a wire (anode) and a work piece (cathode).","In FIG.","1 an arc welding process is schematically illustrated.","The power source 1 is connected to a wire 2 and a work piece 3 to create a closed electric circuit by means of an arc 4 generated in the gap between the wire 2 and the work piece 3 with the purpose of transferring material from the wire 2 to the work piece 3.The welding process may be divided in the following main parts: wire part 5, arc region part 6 and work piece part 7.Furthermore, the arc region part 6 may be divided in subparts, namely an arc-wire interaction region part 8, an arc column region part 9 and an arc-work piece interaction region part 10.All these parts contribute to the characteristics of the welding process and to a certain voltage drop in the electric circuit.","However, in the electrode regions, i.e.","in the arc-wire interaction region part 8 and the arc-work piece interaction region part 10 of the welding process, the voltage drop is very high compared to the voltage drop in the central part of the arc, i.e.","in the arc column region part 9.In FIG.","2 the voltage distribution along the arc, or in other words in the gap between the wire 2 and the work piece 3, is schematically illustrated.","It is indicated that the voltage drops highly in the regions near to the wire and near to the work piece.","This means that the regions situated near to the wire and near to the work piece are important parts in the welding process and have strong influence on the weld joint resulting from the welding operation.","Furthermore, the voltage distribution in the gap is very difficult to determine by direct measuring of the arc whereas the voltage drops due to the resistances and inductances of the conduits 11 in the circuit and of the power source 1 itself are easily determined by conventional measurements.","Thus, there is a need of a physical model for describing the phenomena of the arc to predict the behaviour of the welding process.","However, as already indicated the wire 2 and work piece 3 strongly interact with the arc 4 and influence the welding process.","Thus, good physical models of these parts are also needed to describe the welding process in its entirety.","The main idea of the invention is to obtain the value of at least one welding parameter by means of a theoretical model and use said at least one welding parameter in operating an arc welding equipment and/or in simulating an arc welding process and/or in predicting the quality of a weld obtained from an arc welding operation.","This is performed by dividing the welding process into parts in a theoretical model and letting each of these welding process parts and the welding parameters associated therewith be represented by a model component.","The components are then put in an electric circuit model together with a model power source with the purpose of calculating at least one electric circuit model parameter related to said at least one welding parameter from the electric circuit model.","The components may be resistive and/or inductive components, but also other electric elements than pure resistors and inductors may be included in the electric circuit model.","Furthermore, the model components may be connected in series and/or in parallel.","In FIG.","3 an example of an electric circuit model 12 including model components 13 and a model power source 14 is illustrated.","In this electric circuit model 12 the wire part 5 is represented by a resistor, Rw, the work piece part 7 by a resistor, Rwp, and the arc part 6 is represented by a resistor, Ra.","In addition to these resistances it is necessary to define a model resistance R0 and a model inductance L0 corresponding to the resistance and the inductance from the power source 1 itself and the cables 11 in the real welding circuit.","The circuit model resistance R0 and the inductance L0 are well known properties, or may be chosen, and for the simplification below these are treated as a single component (RL)rest.","(In this component also other resistances, such as a contact resistance between the wire and a nozzle holding the wire, may be included).","Although the electric circuit model 12 illustrated in FIG.","3 comprises four separate components: Rw, Rwp, Ra and (RL)rest, in accordance with the invention the welding process may be divided into two or more parts, but at least two separate parts, of which at least one part represents the arc or a portion thereof present in the welding process.","In the case of two separate model components 13 resistances in the circuit, with the exception of the resistance Ra from the arc region part 6, are suitably included in (RL)rest.","Of course higher dissolution, i.e.","more model components 13 representing different parts in the welding process, gives a more precise description of the arc welding process and also the possibility to obtain information about more welding parameters and to take more characteristics of the welding process into consideration.","However, it is also possible to use different physical models representing subparts of a part of the welding process, the arc region part 6 for instance, which are based on the different characteristics of the part in different regions, for example the arc-wire interaction region 8, the arc column region 9 and/or the arc-work piece interaction region 10, resulting in one common model component such as a resistance Ra.","In this way different parts of the welding process may be described without using a separate model component 13 for each part.","In addition it is also possible to use input values from other parts of the welding process when the model component is calculated.","By using Kirchoff's Voltage Laws at least one electric circuit model parameter may be calculated from the electric circuit model 12.For example, if the voltage supply 14 and the other components 13 are known, the current may be calculated or vice versa for a given current an unknown model component may be calculated.","The model components Rw, Ra and Rwp in FIG.","3 and other such components may be calculated from one or more of the physical models of the wire, arc and the work piece and of their interaction with each other.","Below, examples of physical models are given.","In FIG.","4 the invention and some of its applications are schematically illustrated.","The device according to the invention comprises an arc welding equipment 21 to be used in a welding operation for creating a weld joint.","The welding equipment 21 are suitably an automated or robotic arc welding equipment which may be adjusted by varying at least one welding parameter, such as supply voltage, shielding gas flow rate, welding speed, torque angle, wire extension, wire feed rate etc.","Furthermore, the device includes an arrangement 22 for controlling the operation of the arc welding equipment 21.The control arrangement 22 comprises a means 23, for example a processor and computer programs and electric components/circuits thereto, adapted to determine the value of said at least one welding parameter while using a theoretical model 24 of the welding process, and a member 25, for example any kind of control equipment which is able to communicate with the welding equipment and said means and thus transfer a signal representing the welding parameter value to the welding equipment, for adjusting the welding equipment 21.The adjustment may be performed prior to welding and/or on line during the welding.","The means 23 is adapted to use said theoretical model 24, which comprises at least two separated parts 26, 27, of which at least one part represents the arc or portion thereof, corresponding to different parts of the welding process.","Each model part 26, 27 is represented by a model component 13, see also FIG.","3, and these model components 13 together with the model power source 14 are included in the electric circuit model 12.The means 23 is adapted to calculate at least one electric circuit model parameter 13 from the electric circuit model 12, and to use the value of this parameter related to said at least one welding parameter, for said control of the welding equipment 21 through adjusting of said at least one welding parameter of the welding process by said member 25.As indicated in FIG.","4 the means 23 may be used without the member 25 and the arc welding equipment 21 for other purposes than direct controlling of an arc welding equipment.","Thus, such a means may be used for simulation 28 of any arc welding process and/or for prediction of the quality 29 of a weld resulting from an arc welding operation.","Furthermore, said means 23 and/or the control arrangement 22 may be used in performing any of the methods described herein.","In FIG.","5 a block diagram of the method for controlling an arc welding equipment used in a welding operation and adjustable by varying at least one welding parameter value is illustrated.","The method comprises the steps of determining 30 said at least one welding parameter value while using 31 a theoretical model of the welding process associated with the welding operation, controlling 32 the operation of the welding equipment and the welding process associated therewith by using said at least one welding parameter value for adjusting the welding equipment, where the determining comprises the steps of dividing 33 the welding process into at least two separate parts, of which at least one part represents the arc or a portion thereof, in the theoretical model, representing 34 each of said at least two welding process parts and the welding parameter/parameters associated therewith by a model component, putting 35 the model components and the model power source in an electric circuit model, and calculating 36 at least one electric circuit model parameter related to said at least one welding parameter from the electric circuit model.","In FIG.","6 a block diagram of the method for simulating an arc welding process is illustrated.","The method comprises the steps of inputting 40 data into a theoretical model representing the welding process, determining 41 at least one welding parameter value of the welding process while using 42 the theoretical model and the data input, with the purpose to simulate 43 the welding process, where the determining comprises the steps of dividing 44 the welding process into at least two separate parts, of which at least one part represents the arc or a portion thereof, in the theoretical model, representing 45 each of said at least two welding process parts and the welding parameter/parameters associated therewith by a model component, putting 46 the model components and a model power source in an electric circuit model, and calculating 47 at least one electric circuit model parameter related to said at least one welding parameter from the electric circuit model.","In FIG.","7 a block diagram of the method for predicting the quality of a weld obtained from an arc welding operation is illustrated.","The method comprises the steps of inputting 50 data into a theoretical model representing the welding process associated with the welding operation, determining 51 at least one property of the weld related to at least one welding parameter value of the welding process while using 52 the theoretical model and the data input, where the determining comprises the steps of dividing 53 the welding process into at least two separate parts, of which at least one part represents the arc or a portion thereof, in the theoretical model, representing 54 each of said at least two welding process parts and the welding parameter/parameters associated therewith be represented by a model component, putting 55 the model components and the model power source in an electric circuit model, and calculating 56 at least one electric circuit model parameter related to said at least one welding parameter, and related to said at least one property of the weld, from the electric circuit model.","The methods may be used for different objects such as for predicting the metal droplet formation in an arc welding operation for a given set of welding parameters, for predicting the metal transfer mode in an arc welding operation for a given set of welding parameters, for calculating at least one welding parameter value required for obtaining a particular metal transfer mode in a welding operation and/or for calculating at least one default setting value to be used for adjusting an automatic arc welding equipment prior to welding.","The prediction of quality may be used for example for prediction of any mechanical property of a weld joint for a given input, i.e.","set of welding parameters, or in the reverse direction for determining one or more welding parameter values for a given input, i.e.","any mechanical property of a weld joint.","An important application of the methods and the device according to the invention is the prediction of the characteristics of a certain welding process, and as mentioned above the prediction of the metal transfer mode present during an arc welding operation, which makes it possible to predict the transfer mode as a function of a certain welding parameter, such as wire feed speed or current, and thus to predict the transition region between so called short arc and spray arc mode.","In different embodiments of the methods the step of dividing the welding process may include dividing of the welding process so that the arc-wire interaction region part and/or the arc region part and/or the arc column region part and/or the wire part and/or the work piece part is separately represented by one of said model components 13 in the electric circuit model 12.In embodiments of the methods using an electric circuit model 12 according to FIG.","3 said methods comprise the steps of dividing the welding process so that the arc region part 6, the wire part 5, the work piece part 7 (and the part corresponding to (RL)rest) each is separately represented by one of said model components 13 in the electric circuit model 12.Furthermore, in a preferred embodiment described more in detail below, said model component, Ra, representing the arc region part is determined by means of information obtained through calculation from physical models of the arc column region part 9, the arc-wire interaction region part 8, and the arc-work piece interaction region part 10.In FIGS.","8, 9, 10 and 11 an example of a physical model, including submodels, based on energy, mass and momentum equations is schematically illustrated.","As indicated in FIG.","8 the wire, the arc, the work piece, the arc-wire interaction and the arc-work piece interaction are taken into account.","In FIG.","9 the arc column 60 is assumed to be a cylindric conductor with a mean diameter 61 and an arc expansion transition zone 62 near to the wire 63.In FIG.","10 the arc-wire interaction region part of the welding process is divided into a first zone 64 nearest to the wire, called the space charge zone, and a second zone 65 nearest to the arc column, called the ionization zone, for describing the properties of the zones through different physical submodels.","Also the arc-work piece interaction region part of the welding process in FIG.","11 is divided into a first zone 66 nearest to the work piece, called space charge zone, and a second zone 67 nearest to the arc column, called the ionization zone, for describing the properties of the zones through different physical submodels.","Below follows a more detailed description of possible physical models which may be used in connection with the invention.","PHYSICAL MODELS Arc Model As already mentioned the welding process may be divided into different parts and the arc region part may be divided in subparts, namely an arc-wire interaction region part, an arc column region part and an arc-work piece interaction region part.","The arc column, described in this chapter, occupies most of the space between the wire and the work piece.","In this region, gradients of thermodynamic variables, such as temperature and pressure, are not too high and the system is in local thermodynamic equilibrium.","The arc model suggested here is based on energy, mass and momentum equations.","The arc current generates Lorentz forces which have an important role in the welding process.","The arc is usually constricted towards the wire tip.","This rises the axial component of the Lorentz force.","Plasma is therefore accelerated towards the work piece.","The electromagnetic forces in the welding process are important both for the stabilization of the arc and for material transfer to the work piece.","To be able to predict some phenomena of welding arcs, thermo-physical properties of shielding gas in the temperature range 300-30000 Kelvin are required.","Due to arc-wire and arc-work piece interactions, the shielding gas is usually contaminated by metallic vapours, which strongly influence the physical properties of the arc.","The structure for the arc model proposed herein is shown in FIG.","8.This structure gives the possibility to consider the effects of the shielding gas, the wire material and the work piece material.","According to the state of art, a detailed two-dimensional simulation of the arc is known today.","However, such a simulation requires long computational time.","This makes it difficult to use such a tool for controlling and monitoring the arc welding process.","In this application instead a fast arc model with the structure illustrated in FIG.","8 and 9 is proposed.","The model is based upon the following considerations: the arc is in local thermodynamic equilibrium, the arc is assumed to be a cylindrical conductor with a uniform temperature and a mean diameter, an arc expansion transition zone between the wire and the arc column is considered, the arc mean temperature and diameter are obtained from the arc energy balance and the Steenbeck minimum principle.","This principle is based on the minimization of the arc power loss, thermo-physical properties are calculated in separate modules and coupled to the arc model, and the arc-wire interaction and the arc-work piece interaction are taken into account.","The energy balance of the arc is given below: LI 2 σ A ︸ dissipation in arc column + ( V an + V ca ) I ︸ dissipation due to anode and cathode falls = A ρ v ( h - h 0 ) ︸ convection + ALU rod ︸ radiation + Q A - A ︸ Heat transfer Arc - Anode + Q A - C ︸ Heat transfer Arc - Cathode where L is the arc length, I denotes the electric current, σ refers to the electric conductivity, A is the cross section area of the arc column, Van and Vca are the voltage drops in the near to anode (wire) region and in the near to cathode (work piece) region, respectively.","ρ is the gas density, h and h0 are the specific enthalpies of the arc and the surroundings, and Urad is the mean radiation density, which all are functions of the arc temperature.","ν denotes the mean gas velocity.","The heat transfer terms for arc-anode QA-A and arc-cathode QA-C are obtained from detailed physical models of the arc-cathode and arc-anode interactions.","Using Ampere's law and integration of Navier-Stokes equations for an incompressible flow in the expansion zone a simple equation for the mean velocity can be deduced as v = C a 1 R Arc π C b μ o 2 ρ ( R Arc 2 R Wire 2 - 1 ) where Rarc and Rwire are the radii of the arc column and wire, respectively, and μ0 is the magnetic field constant.","Constants Ca and Cb depend on the current density profiles on the anode wire and will be derived with the help of CFD simulations and simple experiments.","The equations above together with the minimization of the power loss provide all necessary information such as arc mean temperature, arc radius and arc current-voltage characteristics.","The benefits of the proposed arc model are: the arc model may be executed very fastly and thus it may be used as a part of a software for performing the invention and for example be used together with a model for the arc-wire interaction so that droplet formation and transport may be predicted.","Arc-work Piece Interaction Model Near the electrodes, high gradients of temperature and concentration of particles are present.","These lead to deviations from thermal and chemical equilibrium.","It is well known that the voltage.","drops in the near to wire region and in the near to work piece region are very high and dependent on electrode materials and arc column plasma properties.","Therefore, it is important to take physical properties of the near electrodes phenomena for prediction of an arc welding process into consideration.","In this section the model for the interaction between the arc and the work piece is presented.","The model is based on the following: the region near the work piece is divided into two zones: the ionization zone and the space charge zone, collisions between particles are negligible in the space charge zone, electrons and heavy particles are considered as two fluids with two different temperatures, densities of ions and electrons are equal in the ionization zone, densities of ions and electrons are not equal in the space charge zone, the electron emission mechanism is thermo-field emission, and equations describing the two zones are integrated over the zones.","The inputs to the arc-work piece interaction region model are the work piece material and the plasma properties in the arc column region (e.g.","temperature and pressure).","The outputs of the model are the voltage drop over the arc-work piece interaction region and the heat flux from work piece to arc due to charged particles.","Furthermore, there is a conductive heat flux due to neutral particles.","These data are needed in the global energy balance of the arc column.","The model is adapted in a way so that it may easily be coupled to the arc model described above.","The benefits of the proposed arc-work piece interaction model are: the model may be coupled to the arc column model and/or to the arc-wire interaction region model and allows a more complete description of the arc, the model gives accurate predictions of the heat flux into the work piece which is an important factor that effects the quality of the weld, the model predicts the rate of metal vapours from the work piece, the model may be executed very fastly and thus it is for instance not necessary to decrease the speed of a welding operation when using the model for controlling an arc welding operation, and the model considers explicitly the properties of the work piece material, for example the work function and the thermal conductivity.","For illustrating the influence of the work piece material on the arc welding process, a summary of the physics is presented in the following.","The energy flux from the plasma to the work piece is given by: q = J i [ k ( T i 2 + T e 2 - T w 2 ) + e ( φ c + E i - ϕ eff ) ] + J e ( T c 2 k + ϕ eff e ) - J em ( T w 2 k + ϕ eff e ) - q vap where Je, Jem and Ji are the plasma back-dffused electrons current density, the emitted electrons current density and the ion current density, respectively.","Tw, Te and Ti indicate the temperature of the cathode wall, electrons and ions.","φc and Ei are the voltage drop in the space charge zone and the ionization energy of gas.","φeff is the effective work function, which is defined as the difference between the mean energy of electrons inside the metal and the mean energy of the emitted electrons just outside the metal.","qvap is the energy flux carried away by vaporization.","The energy balance in the ionization zone may be summarized as shown in FIG.","12.This figure shows that a part of the total power density, the Joule effect, received by the electron gas is transferred to the heavy particles by elastic and inelastic collisions.","The other part of energy is lost by radiation and by thermal flow to the cold walls.","According to earlier investigations of the ionization zone, the main losses of electronic energy is due to ionization.","Based on these results, the energy losses due to elastic collisions can be neglected.","The energy balance is simplified by integration over the zone: Jem(2kTw+eφc)+Joule_effect=Je(2kTe+eφc)+eJiE+3.2/kTe The first term on the left hand side is the flux of the electrons brought into the ionization zone by the emitted electrons accelerated in the space charge zone.","The second term of the left hand side presents the work of the electric field over the electrons inside the ionization zone.","The first term on the right hand side is the energy flux carried away by the electrons leaving the ionization for the space charge zone.","eJiE is the losses of the electron energy due to ionization.","Finally, the last term on the right hand side is the energy flux carried away by electrons leaving the ionization zone for the arc column.","To be able to predict the cathode region behaviour, the current densities Je, Jem and Ji are required.","These current densities are obtained by detailed kinetics of particles in the space charge zone.","Here only the results are given.","The ion current density and the back-diffused electron current density are given by J i = n io e k ( T i + T e ) m J e = n io e 4 8 k T e π m e exp ( - φ c e T e k ) According to the thermionic field emission mechanism, the emitted electron current density is given by Richardson-Dushman equation with the Schottky correction.","J em = k 2 4 π m e e h 3 T w 2 exp ( ϕ eff e T w k ) In the above equation, h and me are the Planck's constant and the electron mass, respectively.","The effective work function is connected to the electric field according to: ϕ eff = ϕ 0 - ( E r e 4 πɛ ) where Φ0, ∈ and EC are the work function of the work piece material, the dielectric constant and the electric field strength at the cathode wall.","The electron and ion densities at the edge of the space charge zone can be calculated with good approximation as: n i0 = n r0 = n plasma 0.8 2 + α where α = ( T i k m i D io k r n eq 2 ) 1 2 Di0 and kr are the ion-neutral diffusion coefficient at the top of the equilibrium limit and the recombination rate.","The set of equation presented is closed and can be solved easily to predict cathode region properties or arc-work piece interaction region properties in the arc welding process.","Arc-wire Interaction Region Model In this section the model for the interaction between the arc and the wire (anode region) is presented.","The model is based on the following: the region near the wire is divided into two zones: the ionization zone and the space charge zone, collisions between particles are negligible in the space charge zone, electrons and heavy particles are considered as two fluids with two different temperatures, densities of ions is much less than densities of electrons in space charge zone, temperature of electrons remains constant through anode region, the voltage drop in the ionization zone is negligible, and no electron emission from the wire is assumed.","The inputs to the arc-wire interaction region model are the wire material and the plasma properties in the arc column region (e.g.","temperature and pressure).","The outputs of the model are the voltage drop over the arc-wire interaction region and the heat flux from arc to wire due to charged particles.","These data are needed in the global energy balance of the arc column.","The model is adapted in a way so that it may easily be coupled to the arc model described above.","The benefits of the proposed arc-wire interaction model are: the model may be coupled to the arc column model and/or the arc-work piece interaction region model and allows a more complete description of the arc, the model gives accurate predictions of the heat flux into the wire which is an important factor for describing the welding process, the model may be executed very fastly and thus it is for instance not necessary to decrease the speed of a welding operation when using the model for controlling an arc welding operation, and the model considers explicitly the properties of the wire material, for example the work function.","For illustrating the influence of the wire on the arc welding process, a summary of the physics is presented in the following.","The total energy flux from the plasma to the wire is given by the losses from the arc due to charged particles, thermal conduction and radiation minus the heat power utilized for evaporation and radiation of the wire tip surface Q .","= Q .","el + Q .","cond + Q .","rad ︸ energy flux from arc - Q .","gy , wire - Q .","rad , wire ︸ energy flux from wire tip surface For the energy flux of charged particles, electrons are considered only, since the ion current density flux in the arc-wire interaction region is low compared to the electron current density flux Q .","el = j e ( 5 k b 2 e T e + V an + ϕ ) There is energy flux due to thermal energy of electrons, kinetic energy of electrons resulting from acceleration in electric field and potential energy of electrons after phase transformation on the wire surface (free to bound).","je is the current density of the electrons and Te is the electron temperature in the arc-wire interaction region.","The electron temperature is taken as the mean arc core temperature provided by the arc model.","φ is the nominal work function of the wire surface (known), and Van is the voltage drop over the anode region, which consists mainly of the voltage drop in the space charge zone Van=φan where je is obtained from kinetics je=je(ne,φan) where ne is the electron density in the anode region and is calculated using the equation of state, thermodynamic equilibrium laws and the thermodynamic properties of the arc (temperature, pressure).","For obtaining the voltage drop over the arc-wire interaction region the ion current density ji is taken into account.","The following relation can be derived from kinetics ji=ji(ni,φan) where ni is the ion density in the ionization zone, which is equivalent to the electron density ne.","Knowing the total current density j j=je+ji the voltage drop over the arc-wire interaction region can be obtained.","Thermal conduction from the arc results from the temperature gradient in the anode region acting on ions, and the heat fluxes due to the kinetic energy of ions and electrons in the arc-wire interaction region Q .","cond = λ ∂ T ∂ n + n i v i h i + n e v e h e where vi and ve are the velocities of ions and electrons, respectively, which are given by kinetics at thermodynamic equilibrium.","The radiation power from the arc is the fraction of the power totally radiated by the arc that reaches the wire tip surface {dot over (Q)}rad=∈effσTarc4 where ∈eff is an effective emission coefficient of the arc, which can be estimated from the emission coefficient of the arc column, assuming a cylinder, times the fraction of cylinder surface directed towards the wire.","The energy flux due to evaporation of the wire tip surface is given by the product of evaporation rate and latent heat of the wire material {dot over (Q)}ev,wire={dot over (m)}Δh where the evaporation rates may be expressed as a function of arc column pressure and wire tip surface temperature {dot over (m)}={dot over (m)}(P,Tans) The energy flux due to radiation from the wire tip surface is given by {dot over (Q)}rad,wire=∈σTans4 The only unknown remaining is the wire tip surface temperature Tans.","This temperature can be obtained from the wire model.","A simple estimation is that it is equal to the melting temperature of the wire material.","The set of equations presented above is closed and can be solved to predict the anode region properties or arc-wire interaction region properties in the arc welding process.","Wire Model In the arc welding process, heat transfer from the arc to the wire causes rapid formation of droplets at the wire tip.","Impact of detached and transferred droplets through the arc to the work piece may result in volume deformations and alteration of the mechanical properties of the weld.","In the following, a model is suggested for prediction of the mass, heat and momentum transfer from dripping droplets into the work piece.","The shape of melt metal at wire tip is influenced by time-dependent forces such as gravity, inertia, surface tension, electromagnetic force, the effect of arc pressure, viscous drag by the gas flow.","An extensive description of thermophysical phenomena involved in mechanisms of formation of droplets may be formulated accurately with 2-D computer tools based on a detailed analysis of the plasma-electrode interactions in conditions of local thermodynamic equilibrium of the plasma and laminar flow in the calculation domain.","However, calculations required for these operations may be time-consuming and not acceptable in a user interface software to be used for controlling and/or monitoring the quality of a weld in an welding operation.","A simplified approach to the problem can be to consider the one-dimensional energy balance of a wire dominated by the effects of evaporation, metal transfer from the wire, ohmic heating in the wire by the arc current and current transfer from the plasma.","In a quasisteady state mode, the total energy balance of the wire can be written as: Ev+Em=Eoh+Ec when the heat transfer processes for integrated values over the total volume of the wire stick-out are defined as: E v = ∫ S q ev L ⅆ S due to the evaporation from the surface to the tip of the wire; E m = ρ v 0 Sc ∫ T = 300 T mean C p ⅆ T the heat losses due to metal transfer in the form of droplets detaching from the wire tips; E oh = ∫ V w j 2 σ ⅆ υ the heating in the wire by the arc current, and E e = ∫ S [ j i ( V i - V s + 5 k b 2 e T ) + j e ( ϕ + 5 k b 2 e ) T p ] ⅆ S the heat transferred from the plasma to the droplet at the tip of the wire.","In the above relations, Vw is the total volume of the wire stick-out, S is the total external surface of wire, and Sc=πr2 is the cross-sectional area of the wire at the end of the wire at z=0 (T=300 K) and r is its radius.","The quantities Cp, κ and ρ are the specific heat, the thermal conductivity and density of the wire material at the local wire temperature, respectively.","Tmean is the average droplet temperature at the time of detachment, and v0 is the wire feed speed.","The evaporation rate in g.s−1 at the wire tip is denoted as qev=Pevma/4(makbTa/3)0.5 where Pev and ma are the vapour pressure and theatomic mass of the wire, respectively, while kb is the Boltzmann constant and Ta the temperature of the wire surface.","Other symbols are the following: L, the latent heat of evaporation of the wire material, e, the elementary charge, ji the ion current density from the plasma, je the electron current density from the plasma; the total current density is denoted j; Tp is the temperature immediately in front of the wire, φ is the nominal work function of the wire surface; σ represents the electrical conductivity, Vi is the first ionization potential of the plasma gas and Vs the voltage drop across the sheath in the wire region.","In this approximation, conventional heat conduction from the plasma through neutral particles, conduction through the end of the wire and black body radiation are neglected since their effect can theoretically be shown to be small compared to Ev, Em, Eoh and Ee.","Heat transferred from the plasma to the wire may be expressed S a = j i ( V i - V s + 5 k b 2 e T a ) + j e ( ϕ + 5 k b 2 e T p ) - q ev L By solving the time-dependent energy conservation equation at small time steps, ρ ∂ h ∂ t + ρ v 0 ∂ h ∂ z = ∂ ∂ z ( κ ∂ T ∂ z ) + 5 k b 2 e ( j z C p ∂ h ∂ z ) + j 2 σ + F where F is a term referring to the heat input Sa, from the plasma, the temperature profile of the wire can be obtained, and therefore the molten length at the wire tip can be estimated as a function of time.","Metal transfer in the form of a droplet will occur when the molten wire reaches a critical length Lcrit for a given set of welding parameters.","Estimation of the volume of droplet can be deduced from a dynamic treatment in the wire region using simplified expressions of surface tensions, electromagnetic forces, gravity, arc pressure and viscous drag force around the molten metal at the tip of the wire at each time step.","The initial shape of the electrode at a time t=0 can be taken as cylindrical with diameter equal to that of the wire covered by about a 0.1 cm thick layer of molten liquid at the tip of the wire.","Initial conditions are to correspond to an arc operating with continuously moving wire of undeformed cylindrical shape.","The inputs required for the wire model are the arc current, the wire feed speed, the wire diameter, the initial interelectrode separation distance, the torch angle and the characteristics of the arc.","The latter may be supplied into the model by outputs from the arc model.","The outputs include the detachment frequency, the volume of a detached droplet of molten metal, the average temperature in a droplet and the acceleration supported by the droplet in the axis of the arc column at detachment.","This result will provide information for evaluation of reinforcement of joint for a given gas arc welding process and work piece geometry.","Also available from the model will be the wire stick-out length and resistance, which allow a more accurate calculation of the arc length than from the simplified model described below.","As a first step a simplified model for the wire has been used.","The arc length is obtained from a balance between wire feed rate wfr and wire melting rate ⅆ ⅆ t ( L + τ ⅆ L ⅆ t ) = - w fr + I 2 R an + I δ V δ H where τ is a time constant that delays the change of arc length from change of heat input.","The resistance of the wire stickout Ran is calculated assuming a constant stickout temperature R an = ζ A w ( D - L ) where ζ is the electrical resistivity of the wire, Aw the wire cross-section area, D the distance between the nozzle and the work piece.","L is the arc length, which is the output of this model.","The power input considers both Ohmic heating of the wire stickout and energy transfer from arc to wire.","The wire melting rate is obtained by relating the power input to the wire enthalpy per unit length δH.","The latter accounts for energy release due to electron absorption in anode and thermal energy carried by electrons δ V = ϕ an + 5 k B T e 2 e where φan is the work function of the anode material, kB is the Boltzmann's constant, Te is the electron temperature and e is the electron charge.","Work Piece Model As part of multivariable processes, automated arc welding may be defined by so called indirect welding parameters (IWPs), acting upon a set of material parameters (MPs) with a resulting set of direct welding parameters (DWPs).","IWPs should be chosen appropriately for given MPs.","DWPs are to be explicitly fixed by specifications of e.g.","optimal weld bead width, penetration and appearance.","A basic issue to be considered regarding control for arc welding is to determine what is to be controlled and which parameters are accessible to control actions in the process.","See FIGS.","13 and 14.Quality control functionalities to be implemented in the proposed software could consist of: process stability monitoring, i.e.","high speed tracking by e.g.","timedomain statistical analysis based upon measured and predicted average IWPs, for example arc voltage or current intensity.","It may allow detection of disturbing inputs (anomalies in gun-to-work piece distance, insufficient shielding gas coverage, etc.).","Advantageously, vision sensors and sound probes also may be used, quality assurance of geometry and mechanical properties of weld joints (DWPS) by process parameters setting from fast advanced physical models based on MPs and IWPs.","The models may include detailed arc-work piece interactions from weld pool phenomena (arc efficiency, droplet dripping, gas and impurities diffusion, etc.)","and micro structural criteria (micro phases, residual stress, dislocations, etc.).","One work piece model is illustrated in FIG.","17.In practice, modelling of weld properties is often based on neural networks trained for a particular geometry of joints.","Neural networks are parameterised non-linear regression models, see FIG.","14.They can provide adapted solutions for treatment of intricate multi-variable systems when physical models are difficult to establish or do not exist.","Also a program using neural networks should include mass, heat, and momentum transfers from the arc and dripping of droplets into work piece.","Indeed, these quantities may have considerable consequences on the overall geometry of the weld and its intrinsic quality.","An important component in the elaboration of a neural network is the control of the characteristic magnitudes of the weights (noted wj(i) in FIG.","15) of input parameters.","This operation requires that the network is preliminary trained from a set of known outputs when the weights are systematically changed until a best-fit description of the network is obtained as a function of the inputs.","Among other possibilities for modelling of the work piece, a neural network program may be trained on gravity joints obtained under specified conditions (mild steel, gas shielding, wire speed, wire feed rate, plate thickness, etc) by an arc welding robot.","Solidification of melt during GMA welding process depends on heat transfer conditions in the weld pool, see FIG.","16.Fusion mechanism and characteristics of resultant welds are complex to model analytically.","Neural networks can represent a suitable alternative for a model of a workpiece.","Shape and mechanical properties of welds can strongly be affected by electromagnetic forces, buoyancy forces and surface tension forces that induce flow of molten metal in the weld pool possibly affected by impacts of droplets, vaporization, arc pressure and molecular contamination.","Accurate enough predictions of weld quality by analytical model of those phenomena can be difficult to provide as they call for detailed computation and optimized calculation routines including appropriate numerical methods for moving boundary systems.","Neural networks are powerful parameterised non-linear regression models for which no fundamental knowledge of the system to be modelled is required.","They can be generally described as series of weighted variables (input parameters xi) yj=Σiwi(1)xi+θ(1) to form arguments of non-linear regression transfer functions (hidden variables).","Example: hj=tanh yi Since a one hidden-unit model (hj=h, y=W(2)H+θ(2) output parameter) may not be sufficiently flexible, further degrees of non-linearity can be introduced by combining several non-linear regression transfer functions (output parameter y) yj=Σjwj(2)hj+θ(2) hence permitting neural networks to capture almost arbitrarily non-linear relationships.","The capability of neural networks to handle complexity is much related to the number of regression transfer functions, i.e.","the number of hidden layers.","An important component in the elaboration of a neural network is the control of the characteristic magnitudes of the weights (wi(j)) of input quantities.","This operation requires that the network is preliminary trained from a set of known outputs when the weights are systematically changed until a best-fit description of the network is obtained as a function of the inputs.","Numerical procedures can be developed to avoid overfitting difficulties.","In some cases, error estimates can also be calculated to define fitting uncertainty by considering probability distributions of sets of weights instead of using a unique one.","A neural network may be produced to model effects of heat and mass transfers on weld geometry as part of an Arc Welding End-user Software.","However, as already mentioned, many experimental parameters can influence an arc welding process (for example wire properties, shielding gas properties, power supply, wire feed speed, welding speed, electrode extension, etc.—see figure).","A systematic investigation of merely 10 parameters at only three levels of variation would result in 310, i.e.","about 59.103 possible combinations to explore.","Theoretical knowledge of the arc suggests that the arc current, the welding speed and material properties are mainly responsible for heat and mass transfer into the workpiece.","A GMAW process can be decomposed into separate interdependent modules for, respectively, the electrical circuit, the wire (anode), the arc, for the interactions between the arc and the wire (i.e.","droplet formation), the interactions between the arc and the workpiece, and the workpiece itself (cathode).","The amount of welds to be produced for making a neural network model of the workpiece can hence be reduced to a reasonably small number by successively comparing model results for the diverse modules to a few relevant experimental test cases characteristic of the functionality of the module tested, all critical parameters of other modules being fixed in other respects.","In this aim, intern resistances, inductances and resistances on the welding equipment to be used for experiments should be measured beforehand and recording of process signals during welding (e.g.","arc current and voltage, encoding signal form wire feeding system, etc.)","should be carried out with an acquisition unit.","Validation of module for electrical circuit can be achieved by comparing measured arc current and voltage with predicted values of these quantities at different wire feed speeds and supply voltages for different welding modes (short arc, spray, rapid-arc), all other parameters being kept constant and identical between models and experiments in other respects.","Validation of the wire model for simulation of mass transfer mode (i.e.","droplet dripping) can be achieved in a similar way by further measuring the average mass transfer rate from for several dimensions of wire diameter.","Validation of the arc model can be achieved by varying supply voltage and electrode extension for different shielding gases and wire diameters, all other parameters being kept constant.","Adjustments of each model may have to be implemented stepwise during validation of each module.","Eventually, experiments to be carried out for modelling of complete effects of mass and heat transfer into workpiece simply concern the influence of torch and push/drag angles, welding speed, plate thickness, plate material and joint configuration, all other aspects being supposedly encompassed in the previously validated other routines.","For maximum applicability of the software, experiments shall be carried out on different joints (for example overlap, T-joint) made e.g.","in horizontal-vertical configuration of gravity configuration or other, depending on targeted market for the software.","As input from the arc-dependent models to a neural network model of the workpiece, temperature distributions of electrons and ions, current intensity due to both species and evaporation rate at the workpiece surface will be accessible from a pre-calculated matrix established with the validated models.","Mass, heat and speed of transferring droplets will also be referenced as inputs of the neural network from predictions obtained with the validated model for wire.","The output of the neural network should enable a determination of the weld quality as defined either by geometry or mechanical properties.","A complete version of the software is to include all possible aspects of quality.","When quality is associated to weld shape, the outputs will consist of geometrical dimensions of welds.","A sketch of predicted weld geometry will be displayed or dimensions according to the standard EN-25817 will be listed on end-user interface.","When quality is associated to mechanical properties, the outputs will consist of predicted values of e.g.","yield strength, ultimate tensile strength, Charpy toughness and temper embrittlement."]],"string":"[\n [\n \"Arc welding method\",\n \"A method for controlling an arc welding equipment (21) used in a welding operation and adjustable by varying at least one welding parameter value, comprising the steps of determining said at least one welding parameter while using a theoretical model (24) of the welding process associated with the welding operation, and controlling the operation of the welding equipment and the welding process associated therewith by using said at least one welding parameter for adjusting the welding equipment and a device comprising an arc welding equipment (21) used in a welding operation and adjustable by varying at least one welding parameter and an arrangement (22) for controlling the operation of the arc welding equipment, said control arrangement including a means (23) adapted to determine the value of said at least one welding parameter while using a theoretical model (24) of the welding process, and a member (25) adapted to control the operation of the welding equipment and the welding process associated therewith by using said at least one welding parameter value for adjusting the welding equipment.\"\n ],\n [\n \"1.A method for controlling an arc welding equipment used in a welding operation and adjustable by varying at least one welding parameter value, comprising: determining said at least one welding parameter value while using a theoretical model of the welding process associated with the welding operation, controlling the operation of the welding equipment and the welding process associated therewith by using said at least one welding parameter value for adjusting the welding equipment, dividing the welding process into at least two separate parts, of which at least one part represents the arc or a portion thereof, in the theoretical model, representing each of said at least two welding process parts and the welding parameter/parameters associated therewith by a model component, putting the model components and a model power source in an electric circuit model, and calculating at least one electric circuit model parameter value related to said at least one welding parameter from the electric circuit model.\",\n \"2.A method for simulating an arc welding process, comprising: inputting data into a theoretical model representing the welding process, determining at least one welding parameter value of the welding process while using the theoretical model and the data input, with the purpose to simulate the welding process, dividing the welding process into at least two separate parts, of which at least one part represents the arc or a portion thereof, in the theoretical model, representing each of said at least two welding process parts and the welding parameter/parameters associated therewith by a model component, putting the model components and a model power source in an electric circuit model, and calculating at least one electric circuit model parameter value related to said at least one welding parameter from the electric circuit model.\",\n \"3.A method for predicting the quality of a weld obtained from an arc welding operation, comprising: inputting data into a theoretical model representing the welding process associated with the welding operation, determining at least one property of the weld related to at least one welding parameter of the welding process while using the theoretical model and the data input, dividing the welding process into at least two separate parts, of which at least one part represents the arc or a portion thereof, in the theoretical model, representing each of said at least two welding process parts and the welding parameter/parameters associated therewith by a model component, putting the model components and a model power source in an electric circuit model, and calculating at least one electric circuit model parameter value related to said at least one welding parameter, and related to said at least one property of the weld, from the electric circuit model.\",\n \"4.The method according to claim 1, further comprising: dividing the welding process so that the arc-wire interaction region part of the welding process is separately represented by one of said model components in the electric circuit model.\",\n \"5.The method according to claim 1, further comprising: dividing the welding process so that the arc-work piece interaction region part of the welding process is separately represented by one of said model components in the electric circuit model.\",\n \"6.The method according to claim 1, further comprising: dividing the welding process so that the arc-wire interaction region part of the welding process is separately represented by one of said model components in the electric circuit model, and/or dividing the welding process so that the arc-work piece interaction region part of the welding process is separately represented by one of said model components in the electric circuit model.\",\n \"7.The method according to claim 1, further comprising: dividing the welding process so that the arc region part of the welding process is separately represented by one of said model components in the electric circuit model.\",\n \"8.The method according to claim 7, further comprising: determining said model component representing the arc region part by means of information obtained through calculations from a physical model of the arc column region part of the welding process.\",\n \"9.The method according to claim 7, further comprising: determining said model component representing the arc region part by means of information obtained through calculations from a physical model of the arc-wire interaction region part of the welding process.\",\n \"10.The method according to claim 4, further comprising dividing the arc-wire interaction region part of the welding process into a first zone nearest to the wire and a second zone nearest to the arc column for describing the properties of the zones through different physical submodels.\",\n \"11.The method according to claim 7, further comprising: determining said model component representing the arc region part by means of information obtained through calculations from a physical model of the arc-work piece interaction region part of the welding process.\",\n \"12.The method according to claim 5, further comprising: dividing the arc-work piece interaction region part of the welding process into a first zone nearest to the work piece and a second zone nearest to the arc column for describing the properties of the zones through different physical submodels.\",\n \"13.The method according to claim 7, further comprising: determining said model component representing the arc region part by means of said physical model describing the arc by at least one equation independent of the extension in space of the arc.\",\n \"14.The method according to claim 13, further comprising: determining said model component representing the arc region part by means of said physical model describing the arc by said at least one equation only dependent on the time.\",\n \"15.The method according to claim 1, further comprising: dividing the welding process so that the arc column region part of the welding process is separately represented by one of said model components in the electric circuit model.\",\n \"16.The method according to claim 1, further comprising: dividing the welding process so that the wire part of the welding process is separately represented by one of said model components in the electric circuit model.\",\n \"17.The method according to claim 1, further comprising: dividing the welding process so that the work piece part of the welding process is separately represented by one of said model components in the electric circuit model.\",\n \"18.A device comprising an arc welding equipment used in a welding operation and adjustable by varying at least one welding parameter and an arrangement for controlling the operation of the arc welding equipment, said control arrangement including a means adapted to determine the value of said at least one welding parameter while using a theoretical model of the welding process, and a member adapted to control the operation of the welding equipment and the welding process associated therewith by using said at least one welding parameter value for adjusting the welding equipment, wherein the means is adapted to use said theoretical model comprising at least two separate parts, of which at least one part represents the arc or a portion thereof, corresponding to different parts of the welding process, each model part being represented by a model component, said model components together with a model power source being included in an electric circuit model and that said means is adapted to calculate at least one electric circuit model parameter value related to said at least one welding parameter from the electric circuit model for said control by said member.\",\n \"19.The device according to claim 18, wherein the means is adapted to use said theoretical model having the arc-wire interaction region part of the welding process separately represented by one of said model components in the electric circuit model.\",\n \"20.The device according to claim 17, wherein the means is adapted to use said theoretical model having the arc-work piece interaction region part of the welding process separately represented by one of said model components in the electric circuit model.\",\n \"21.The device according to claim 18, wherein the means is adapted to use said theoretical model having the arc-wire interaction region part of the welding process separately represented by one of said model components in the electric circuit model and/or the arc-work piece interaction region part of the welding process separately represented by one of said model components in the electric circuit model.\",\n \"22.The device according to claim 18, wherein the means is adapted to use said theoretical model having the arc region part of the welding process separately represented by one of said model components in the electric circuit model.\",\n \"23.The device according to claim 22, wherein the means is adapted to use said theoretical model having information obtained through calculations from a physical model of the arc column region part of the welding process to be used in determining said model component representing the arc region part.\",\n \"24.The device according to claim 22, wherein the means is adapted to use said theoretical model having information obtained through calculations from a physical model of the arc-wire interaction region part of the welding process to be used in determining said model component representing the arc region part.\",\n \"25.The device according to claim 19, wherein the means is adapted to use said theoretical model having the arc-wire interaction region part of the welding process divided into a first zone nearest to the wire and a second zone nearest to the arc column for describing the properties of the zones through different physical submodels.\",\n \"26.The device according to claim 22, wherein the means is adapted to use said theoretical model having information obtained through calculations from a physical model of the arc-work piece interaction region part of the welding process to be used in determining said model component representing the arc region part.\",\n \"27.The device according to claim 20, wherein the means is adapted to use said theoretical model having the arc-work piece interaction region part of the welding process divided into a first zone nearest to the work piece and a second zone nearest to the arc column for describing the properties of the zones through different physical submodels.\",\n \"28.The device according to claim 22, wherein the means is adapted to use said model component determined by means of said physical model describing the arc by at least one equation independent of the extension in space of the arc.\",\n \"29.The device according to claim 28, wherein the means is adapted to use said model component determined by means of said physical model describing the arc by said at least one equation only dependent on the time.\",\n \"30.The device according to claim 18, wherein the means is adapted to use said theoretical model having the arc column region part of the welding process separately represented by one of said model components in the electric circuit model.\",\n \"31.The device according to claim 18, wherein the means is adapted to use said theoretical model having the wire part of the welding process separately represented by one of said model components in the electric circuit model.\",\n \"32.The device according to claim 18, wherein the means is adapted to use said theoretical model having the work piece part of the welding process separately represented by one of said model components in the electric circuit model.\",\n \"33.A use of a method according to claim 2, for predicting the metal droplet formation in an arc welding operation for a given set of welding parameters.\",\n \"34.A use of a method according to claim 2, for predicting the metal transfer mode in an arc welding operation for a given set of welding parameters.\",\n \"35.A use of a method according to claim 1, for calculating at least one welding parameter value required for obtaining a particular metal transfer mode in a welding operation.\",\n \"36.A use of a method according to claim 1, for calculating at least one default setting value to be used for adjusting an automatic arc welding equipment prior to welding.\",\n \"37.A computer program product for operating a method according to claim 1.38.A computer program product comprising code means and/or software code portions for enabling a processor to perform the steps of claim 1.39.A computer program product for operating a device according to claim 18.40.A computer program product according to claim 37, provided at least partially through a network such as the Internet.\",\n \"41.A computer readable medium containing a computer program product according to claim 37.\"\n ],\n [\n \"<SOH> FIELD OF THE INVENTION AND PRIOR ART <EOH>The present invention relates to a method for controlling an arc welding equipment used in a welding operation and adjustable by varying at least one welding parameter value, which method comprises the steps of: determining said at least one welding parameter value while using a theoretical model of the welding process associated with the welding operation and controlling the operation of the welding equipment and the welding process associated therewith by using said at least one welding parameter value for adjusting the welding equipment, to a method for simulating an arc welding process which method comprises the steps of: inputting data into a theoretical model representing the welding process and determining at least one welding parameter value of the welding process while using the theoretical model and the data input, with the purpose to simulate the welding process, and to a method for predicting the quality of a weld obtained from an arc welding operation which method comprises the steps of: inputting data into a theoretical model representing the welding process associated with the welding operation and determining at least one property of the weld related to at least one welding parameter of the welding process while using the theoretical model and the data input.\",\n \"The invention also relates to a device comprising an arc welding equipment used in a welding operation and adjustable by varying at least one welding parameter and an arrangement for controlling the operation of the arc welding equipment, said control arrangement including a means adapted to determine the value of said at least one welding parameter while using a theoretical model of the welding process, and a member adapted to control the operation of the welding equipment and the welding process associated therewith by using said at least one welding parameter for adjusting the welding equipment.\",\n \"Furthermore, the invention relates to uses of the methods and to computer program products according to claims enclosed.\",\n \"When work pieces are joined by arc welding the quality of the welded joints is strongly dependent upon the parameters set during the arc welding operation.\",\n \"Examples of these welding parameters are welding current, supply voltage, type of shielding gas, shielding gas flow rate, welding speed, torch angle, electrode (wire) extension, wire feed rate, and other properties of the wire and of the work piece, such as dimensions and materials.\",\n \"These welding parameters interact with each other in the welding process in a complex manner and influence the properties, i.e.\",\n \"the geometry or type of weld, metallurgy and the mechanical strength of the weld resulting from the welding operation.\",\n \"More precisely, the quality of a weld joint depends strongly on the metal transfer mode present during the welding.\",\n \"In other words, the way metal is transferred from the wire tip (for example the anode) to the work piece (for example the cathode) during the welding has a very strong effect on the final properties and the quality of the weld joint.\",\n \"There is of course a need to know how different sets of the adjustable parameters will effect the properties of the weld joint, so that the parameters may be adjusted in a way resulting in the weld desired.\",\n \"One way is to attempt to predict the metal transfer mode based on the knowledge about the influence of the different parameters.\",\n \"However, according to the state of art the metal transfer mode is mainly determined through empirical methods.\",\n \"The metal transfer mode may be identified through the noise generated by the welding process by people with a very long experience in arc welding.\",\n \"Inspection of the weld joint after finishing the welding operation may also give clues to which metal transfer mode that was present during the welding operation.\",\n \"Thus, at present, the settings of important welding process parameters used in robotic arc welding are found out experimentally by trial and error for every single welding task or welding condition, which is both time consuming and costly.\",\n \"For example, in case of using parameters resulting in an undesired metal transfer mode the welded joint may be of a poor quality and the work piece may be destroyed.\",\n \"One reason to the lack of knowledge about how the welding parameters effect the welding process in general, and particularly the arc, and about the relations between the welding parameters used and the quality of the weld joint, is the difficulty in making measurements of the arc, or in the vicinity of the arc, and in the near to arc regions close to the wire and the work piece.\",\n \"Thus, as an undesired consequence it is very difficult to design a control system based upon direct measurements of the arc parameters for controlling an arc welding equipment.\",\n \"The fact is that automated or robotic arc welding are used in quite few of the arc welding applications performed throughout the world today, since the difficulty in predicting the result of the welding operation and the properties of welding process associated thereto for a given set of welding parameters, together with proportionately small series of work pieces, makes it more cost efficient to perform the work manually by workers skilled in welding.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>A first object of the invention is to provide a method for controlling an arc welding equipment, which method enabling that the arc welding equipment may be controlled during a welding operation by adjusting at least one welding parameter determined without the need of measurements of the welding process or repeated welding experiments prior to welding.\",\n \"The first object is obtained according to the invention by providing such a method, referred to in the introduction, which comprises the steps of: dividing the welding process into at least two separate parts, of which at least one part represents the arc or a portion thereof, in the theoretical model, representing each of said at least two welding process parts and the welding parameter/parameters associated therewith by a model component, putting the model components and a model power source in an electric circuit model, and calculating at least one electric circuit model parameter related to said at least one welding parameter from the electric circuit model.\",\n \"By such a method it is possible to determine at least one welding parameter value, such as the welding current or voltage supply, wire feed rate, wire extension etc., and to use this welding parameter value for controlling the arc welding equipment in accordance with the present conditions to obtain a weld with the properties desired.\",\n \"A second object of the invention is to provide a method for simulating an arc welding process without the need of measurements of the corresponding real welding process or repeated welding experiments.\",\n \"The second object is obtained according to the invention by providing such a method, referred to in the introduction, which comprises the steps of: dividing the welding process into at least two separate parts, of which at least one part represents the arc or a portion thereof, in the theoretical model, representing each of said at least two welding process parts and the welding parameter/parameters associated therewith by a model component, putting the model components and a model power source in an electric circuit model, and calculating at least one electric circuit model parameter related to said at least one welding parameter from the electric circuit model.\",\n \"By this simulation method it is possible to obtain at least one welding parameter value, and/or the present metal transfer mode, for a given set of other welding parameters in the simulation, and for example this simulation method may be used for understanding and designing of real welding processes and for default setting of welding parameters of arc welding equipment in different arc welding operations.\",\n \"A third object of the invention is to provide a method for predicting the quality of a weld obtained from an arc welding operation without the need of repeated welding experiments.\",\n \"The third object is obtained according to the invention by providing such a method, referred to in the introduction, which comprises the steps of: dividing the welding process into at least two separate parts, of which at least one part represents the arc or a portion thereof, in the theoretical model, representing each of said at least two welding process parts and the welding parameter/parameters associated therewith by a model component, putting the model components and a model power source in an electric circuit model, and calculating at least one electric circuit model parameter related to said at least one welding parameter, and related to said at least one property of the weld, from the electric circuit model.\",\n \"By such a method it is possible to predict one or more mechanical properties of a weld resulting from an arc welding operation by determining one or more welding parameters required for the quality desired and use this information in real arc welding operations.\",\n \"It is possible to use the method for predicting the quality of a weld for a given set of welding parameters as well as for determining one or more welding parameters required for obtaining certain mechanical properties of the weld.\",\n \"According to a preferred embodiment of the invention one or more of said methods comprise the step of: dividing the welding process so that the arc-wire interaction region part of the welding process is separately represented by one of said model components in the electric circuit model.\",\n \"By such a method, together with a suitable physical model, important information is obtained because it is possible to take important factors in the welding process, such as the voltage drop over the region near to anode and the heat flux from arc to anode, into consideration in determining said at least one welding parameter value.\",\n \"In the arc-wire interaction region of the welding process, high gradients of different quantities such as temperature and concentration of particles are present, and the voltage drop in this region is very high.\",\n \"Thus, this region gives an important contribution to the characteristics of the welding process.\",\n \"According to another preferred embodiment of the invention one or more of said methods comprise the step of: dividing the welding process so that the arc-work piece interaction region part of the welding process is separately represented by one of said model components in the electric circuit model.\",\n \"By such a method, together with a suitable physical model, important information is obtained because it is possible to take important factors in the welding process, such as the voltage drop over the region near to cathode and the heat flux from arc to cathode, into consideration in determining said at least one welding parameter value.\",\n \"In the arc-work piece interaction region of the welding process, high gradients of different quantities such as temperature and concentration of particles are present, and the voltage drop in this region is very high.\",\n \"Thus, this region gives an important contribution to the characteristics of the welding process.\",\n \"According to another preferred embodiment of the invention one or more of said methods comprise the step of: dividing the welding process so that the arc region part of the welding process is separately represented by one of said model components in the electric circuit model.\",\n \"By such a method, and particularly together with an embodiment comprising the step of: determining said model component representing the arc region part by means of information obtained through calculations from a physical model of the arc column region part of the welding process, important information is obtained such as arc mean temperature, temperature near to the wire, temperature near to the work piece, arc radius and arc current-voltage characteristics, and it is also possible to take the thermophysical properties of a shielding gas present in the arc region into consideration in determining said at least one welding parameter value.\",\n \"According to another preferred embodiment of the invention one or more of said methods comprise the step of: determining said model component representing the arc region part by means of information obtained through calculations from a physical model of the arc-wire interaction region part and/or of the arc-work piece interaction region part of the welding process, important information is obtained because it is possible to take important factors in the welding process, such as the voltage drop over the region near to anode/cathode and the heat flux from arc to anode/cathode, into consideration in determining said at least one welding parameter value.\",\n \"In the arc-wire interaction region and in the arc-work piece interaction region of the welding process, high gradients of different quantities such as temperature and concentration of particles are present, and the voltage drop in these regions is very high.\",\n \"Thus, these regions give an important contribution to the characteristics of the welding process.\",\n \"According to another preferred embodiment of the invention one or more of said methods comprise the step of: determining said model component representing the arc region part by means of said physical model describing the arc by at least one equation independent of the extension in space of the arc.\",\n \"By such a method, and particularly together with an embodiment comprising the step of: determining said model component representing the arc region part by means of said physical model describing the arc by said at least one equation only dependent on the time, the model component and thus said at least one welding parameter value, may be determined fast and with proportionally short computational time, which makes the method suitable to be used for example in controlling an arc welding equipment on line.\",\n \"According to another preferred embodiment of the invention one or more of said methods comprise the step of: dividing the welding process so that the arc column region part of the welding process is separately represented by one of said model components in the electric circuit model.\",\n \"By such a method, together with a suitable physical model, important information is obtained such as arc mean temperature, temperature near to the wire, temperature near to the work piece, arc radius and arc current-voltage characteristics, and it is also possible to take the thermophysical properties of a shielding gas present in the arc column region into consideration in determining said at least one welding parameter.\",\n \"According to another preferred embodiment of the invention said methods comprise the step of: dividing the welding process so that the wire part of the welding process is separately represented by one of said model components in the electric circuit model.\",\n \"By such a method, together with a suitable physical model, important information is obtained such as metal transfer mode, dipping frequency etc, and it is also possible to take material properties of the wire into consideration in determining the value of said at least one welding parameter.\",\n \"According to another preferred embodiment of the invention said methods comprise the step of: dividing the welding process so that the work piece part of the welding process is separately represented by one of said model components in the electric circuit model.\",\n \"By such a method, together with a suitable physical model, important information is obtained about quality features such as weld joint profile, penetration profile etc, and it is also possible to take material properties of the work piece into consideration in determining the value of said at least one welding parameter.\",\n \"A fourth object of the invention is to provide a device which has an arrangement for controlling the arc welding equipment during a welding operation by adjusting at least one welding parameter determined without the need of measurements of the welding process or repeated welding experiments prior to welding.\",\n \"The fourth object is obtained according to the invention by providing such a device, referred to in the introduction, in which the means included in the control arrangement is adapted to use said theoretical model comprising at least two separate parts, of which at least one part represents the arc or a portion thereof, corresponding to different parts of the welding process, each model part being represented by a model component, said model components together with a model power source being included in an electric circuit model, and adapted to calculate at least one electric circuit model parameter related to said at least one welding parameter from the electric circuit model for said control by said member.\",\n \"Thus, by the means according to the invention it is possible to determine how to set at least one welding parameter value, such as the welding current or supply voltage, wire feed rate, wire extension etc., with the purpose of optimizing the welding process under the present conditions, and use this welding parameter value by said member for adjusting the arc welding equipment in accordance with the present conditions to obtain a weld with the properties desired.\",\n \"According to a preferred embodiment of the invention said means is adapted to use said theoretical model having the arc-wire interaction region part of the welding process separately represented by one of said model components in the electric circuit model.\",\n \"By such a device, together with a suitable physical model, important information is obtained because it is possible to take important factors in the welding process, such as the voltage drop over the region near to anode and the heat flux from arc to anode, into consideration in determining said at least one welding parameter value.\",\n \"In the arc-wire interaction region of the welding process, high gradients of different quantities such as temperature and concentration of particles are present, and the voltage drop in this region is very high.\",\n \"Thus, this region gives an important contribution to the characteristics of the welding process.\",\n \"According to another preferred embodiment of the invention said means is adapted to use said theoretical model having the arc-work piece interaction region part of the welding process separately represented by one of said model components in the electric circuit model.\",\n \"By such a device, together with a suitable physical model, important information is obtained because it is possible to take important factors in the welding process, such as the voltage drop over the region near to cathode and the heat flux from arc to cathode, into consideration in determining said at least one welding parameter value.\",\n \"In the arc-work piece interaction region of the welding process, high gradients of different quantities such as temperature and concentration of particles are present, and the voltage drop in this region is very high.\",\n \"Thus, this region gives an important contribution to the characteristics of the welding process.\",\n \"According to another preferred embodiment of the invention said means is adapted to use said theoretical model having the arc region part of the welding process separately represented by one of said model components in the electric circuit model.\",\n \"By such a device, and particularly together with an embodiment of the invention in which the means is adapted to use said theoretical model having information obtained through calculations from a physical model of the arc column region part of the welding process to be used in determining said model component representing the arc region part, important information is obtained such as arc mean temperature, temperature near to wire, temperature near to work piece, arc radius and arc current-voltage characteristics, and it is also possible to take the thermo-physical properties of a shielding gas present in the arc region into consideration in determining said at least one welding parameter.\",\n \"According to another preferred embodiment of the invention said means is adapted to use said theoretical model having information obtained through calculations from a physical model of the arc-wire interaction region part and/or of the arc-work piece interaction region part of the welding process to be used in determining said model component representing the arc region part, important information is obtained because it is possible to take important factors in the welding process, such as the voltage drop over the region near to anode/cathode and the heat flux from arc to anode/cathode, into consideration in determining said at least one welding parameter value.\",\n \"In the arc-wire interaction region and in the arc-work piece interaction region of the welding process, high gradients of different quantities such as temperature and concentration of particles are present, and the voltage drop in these regions is very high.\",\n \"Thus, these regions give an important contribution to the characteristics of the welding process.\",\n \"According to another preferred embodiment of the invention in which said means is adapted to use said model component determined by means of said physical model describing the arc by at least one equation independent of the extension in space of the arc, and particularly together with an embodiment of the invention in which said means is adapted to use said model component determined by means of said physical model describing the arc by said at least one equation only dependent on the time, the model component and thus said at least one welding parameter value, may be determined fast and with proportionally short computational time, which makes the device very suitable to be used in controlling the arc welding equipment on line.\",\n \"According to another preferred embodiment of the invention said means is adapted to use said theoretical model having the arc column region part of the welding process separately represented by one of said model components in the electric circuit model.\",\n \"By such a device, together with a suitable physical model, important information is obtained such as arc mean temperature, temperature near to wire, temperature near to work piece, arc radius and arc current-voltage characteristics, and it is also possible to take the thermo-physical properties of a shielding gas present in the arc column region into consideration in determining said at least one welding parameter.\",\n \"According to another preferred embodiment of the invention said means is adapted to use said theoretical model having the wire part of the welding process separately represented by one of said model components.\",\n \"By such a device, together with a suitable physical model, important information is obtained such as metal transfer mode, dipping frequency etc, and it is also possible to take material properties of the wire into consideration in determining the value of said at least one welding parameter.\",\n \"According to another preferred embodiment of the invention said means is adapted to use said theoretical model having the work piece part of the welding process separately represented by one of said model components.\",\n \"By such a device, together with a suitable physical model, important information is obtained about quality features such as weld joint profile, penetration profile etc, and it is also possible to take material properties of the work piece into consideration in determining the value of said at least one welding parameter.\",\n \"The invention also relates to computer program products according to the claims enclosed.\",\n \"It is obvious that the methods according to the invention defined in the sets of method claims enclosed are well suited to be performed through program instructions from a processor which may be effected by a computer program.\",\n \"Further advantages of and features of the methods and the device, the computer program products, and the uses of the methods appear from the following detailed description and the other claims enclosed herein.\"\n ],\n [\n \"FIELD OF THE INVENTION AND PRIOR ART The present invention relates to a method for controlling an arc welding equipment used in a welding operation and adjustable by varying at least one welding parameter value, which method comprises the steps of: determining said at least one welding parameter value while using a theoretical model of the welding process associated with the welding operation and controlling the operation of the welding equipment and the welding process associated therewith by using said at least one welding parameter value for adjusting the welding equipment, to a method for simulating an arc welding process which method comprises the steps of: inputting data into a theoretical model representing the welding process and determining at least one welding parameter value of the welding process while using the theoretical model and the data input, with the purpose to simulate the welding process, and to a method for predicting the quality of a weld obtained from an arc welding operation which method comprises the steps of: inputting data into a theoretical model representing the welding process associated with the welding operation and determining at least one property of the weld related to at least one welding parameter of the welding process while using the theoretical model and the data input.\",\n \"The invention also relates to a device comprising an arc welding equipment used in a welding operation and adjustable by varying at least one welding parameter and an arrangement for controlling the operation of the arc welding equipment, said control arrangement including a means adapted to determine the value of said at least one welding parameter while using a theoretical model of the welding process, and a member adapted to control the operation of the welding equipment and the welding process associated therewith by using said at least one welding parameter for adjusting the welding equipment.\",\n \"Furthermore, the invention relates to uses of the methods and to computer program products according to claims enclosed.\",\n \"When work pieces are joined by arc welding the quality of the welded joints is strongly dependent upon the parameters set during the arc welding operation.\",\n \"Examples of these welding parameters are welding current, supply voltage, type of shielding gas, shielding gas flow rate, welding speed, torch angle, electrode (wire) extension, wire feed rate, and other properties of the wire and of the work piece, such as dimensions and materials.\",\n \"These welding parameters interact with each other in the welding process in a complex manner and influence the properties, i.e.\",\n \"the geometry or type of weld, metallurgy and the mechanical strength of the weld resulting from the welding operation.\",\n \"More precisely, the quality of a weld joint depends strongly on the metal transfer mode present during the welding.\",\n \"In other words, the way metal is transferred from the wire tip (for example the anode) to the work piece (for example the cathode) during the welding has a very strong effect on the final properties and the quality of the weld joint.\",\n \"There is of course a need to know how different sets of the adjustable parameters will effect the properties of the weld joint, so that the parameters may be adjusted in a way resulting in the weld desired.\",\n \"One way is to attempt to predict the metal transfer mode based on the knowledge about the influence of the different parameters.\",\n \"However, according to the state of art the metal transfer mode is mainly determined through empirical methods.\",\n \"The metal transfer mode may be identified through the noise generated by the welding process by people with a very long experience in arc welding.\",\n \"Inspection of the weld joint after finishing the welding operation may also give clues to which metal transfer mode that was present during the welding operation.\",\n \"Thus, at present, the settings of important welding process parameters used in robotic arc welding are found out experimentally by trial and error for every single welding task or welding condition, which is both time consuming and costly.\",\n \"For example, in case of using parameters resulting in an undesired metal transfer mode the welded joint may be of a poor quality and the work piece may be destroyed.\",\n \"One reason to the lack of knowledge about how the welding parameters effect the welding process in general, and particularly the arc, and about the relations between the welding parameters used and the quality of the weld joint, is the difficulty in making measurements of the arc, or in the vicinity of the arc, and in the near to arc regions close to the wire and the work piece.\",\n \"Thus, as an undesired consequence it is very difficult to design a control system based upon direct measurements of the arc parameters for controlling an arc welding equipment.\",\n \"The fact is that automated or robotic arc welding are used in quite few of the arc welding applications performed throughout the world today, since the difficulty in predicting the result of the welding operation and the properties of welding process associated thereto for a given set of welding parameters, together with proportionately small series of work pieces, makes it more cost efficient to perform the work manually by workers skilled in welding.\",\n \"SUMMARY OF THE INVENTION A first object of the invention is to provide a method for controlling an arc welding equipment, which method enabling that the arc welding equipment may be controlled during a welding operation by adjusting at least one welding parameter determined without the need of measurements of the welding process or repeated welding experiments prior to welding.\",\n \"The first object is obtained according to the invention by providing such a method, referred to in the introduction, which comprises the steps of: dividing the welding process into at least two separate parts, of which at least one part represents the arc or a portion thereof, in the theoretical model, representing each of said at least two welding process parts and the welding parameter/parameters associated therewith by a model component, putting the model components and a model power source in an electric circuit model, and calculating at least one electric circuit model parameter related to said at least one welding parameter from the electric circuit model.\",\n \"By such a method it is possible to determine at least one welding parameter value, such as the welding current or voltage supply, wire feed rate, wire extension etc., and to use this welding parameter value for controlling the arc welding equipment in accordance with the present conditions to obtain a weld with the properties desired.\",\n \"A second object of the invention is to provide a method for simulating an arc welding process without the need of measurements of the corresponding real welding process or repeated welding experiments.\",\n \"The second object is obtained according to the invention by providing such a method, referred to in the introduction, which comprises the steps of: dividing the welding process into at least two separate parts, of which at least one part represents the arc or a portion thereof, in the theoretical model, representing each of said at least two welding process parts and the welding parameter/parameters associated therewith by a model component, putting the model components and a model power source in an electric circuit model, and calculating at least one electric circuit model parameter related to said at least one welding parameter from the electric circuit model.\",\n \"By this simulation method it is possible to obtain at least one welding parameter value, and/or the present metal transfer mode, for a given set of other welding parameters in the simulation, and for example this simulation method may be used for understanding and designing of real welding processes and for default setting of welding parameters of arc welding equipment in different arc welding operations.\",\n \"A third object of the invention is to provide a method for predicting the quality of a weld obtained from an arc welding operation without the need of repeated welding experiments.\",\n \"The third object is obtained according to the invention by providing such a method, referred to in the introduction, which comprises the steps of: dividing the welding process into at least two separate parts, of which at least one part represents the arc or a portion thereof, in the theoretical model, representing each of said at least two welding process parts and the welding parameter/parameters associated therewith by a model component, putting the model components and a model power source in an electric circuit model, and calculating at least one electric circuit model parameter related to said at least one welding parameter, and related to said at least one property of the weld, from the electric circuit model.\",\n \"By such a method it is possible to predict one or more mechanical properties of a weld resulting from an arc welding operation by determining one or more welding parameters required for the quality desired and use this information in real arc welding operations.\",\n \"It is possible to use the method for predicting the quality of a weld for a given set of welding parameters as well as for determining one or more welding parameters required for obtaining certain mechanical properties of the weld.\",\n \"According to a preferred embodiment of the invention one or more of said methods comprise the step of: dividing the welding process so that the arc-wire interaction region part of the welding process is separately represented by one of said model components in the electric circuit model.\",\n \"By such a method, together with a suitable physical model, important information is obtained because it is possible to take important factors in the welding process, such as the voltage drop over the region near to anode and the heat flux from arc to anode, into consideration in determining said at least one welding parameter value.\",\n \"In the arc-wire interaction region of the welding process, high gradients of different quantities such as temperature and concentration of particles are present, and the voltage drop in this region is very high.\",\n \"Thus, this region gives an important contribution to the characteristics of the welding process.\",\n \"According to another preferred embodiment of the invention one or more of said methods comprise the step of: dividing the welding process so that the arc-work piece interaction region part of the welding process is separately represented by one of said model components in the electric circuit model.\",\n \"By such a method, together with a suitable physical model, important information is obtained because it is possible to take important factors in the welding process, such as the voltage drop over the region near to cathode and the heat flux from arc to cathode, into consideration in determining said at least one welding parameter value.\",\n \"In the arc-work piece interaction region of the welding process, high gradients of different quantities such as temperature and concentration of particles are present, and the voltage drop in this region is very high.\",\n \"Thus, this region gives an important contribution to the characteristics of the welding process.\",\n \"According to another preferred embodiment of the invention one or more of said methods comprise the step of: dividing the welding process so that the arc region part of the welding process is separately represented by one of said model components in the electric circuit model.\",\n \"By such a method, and particularly together with an embodiment comprising the step of: determining said model component representing the arc region part by means of information obtained through calculations from a physical model of the arc column region part of the welding process, important information is obtained such as arc mean temperature, temperature near to the wire, temperature near to the work piece, arc radius and arc current-voltage characteristics, and it is also possible to take the thermophysical properties of a shielding gas present in the arc region into consideration in determining said at least one welding parameter value.\",\n \"According to another preferred embodiment of the invention one or more of said methods comprise the step of: determining said model component representing the arc region part by means of information obtained through calculations from a physical model of the arc-wire interaction region part and/or of the arc-work piece interaction region part of the welding process, important information is obtained because it is possible to take important factors in the welding process, such as the voltage drop over the region near to anode/cathode and the heat flux from arc to anode/cathode, into consideration in determining said at least one welding parameter value.\",\n \"In the arc-wire interaction region and in the arc-work piece interaction region of the welding process, high gradients of different quantities such as temperature and concentration of particles are present, and the voltage drop in these regions is very high.\",\n \"Thus, these regions give an important contribution to the characteristics of the welding process.\",\n \"According to another preferred embodiment of the invention one or more of said methods comprise the step of: determining said model component representing the arc region part by means of said physical model describing the arc by at least one equation independent of the extension in space of the arc.\",\n \"By such a method, and particularly together with an embodiment comprising the step of: determining said model component representing the arc region part by means of said physical model describing the arc by said at least one equation only dependent on the time, the model component and thus said at least one welding parameter value, may be determined fast and with proportionally short computational time, which makes the method suitable to be used for example in controlling an arc welding equipment on line.\",\n \"According to another preferred embodiment of the invention one or more of said methods comprise the step of: dividing the welding process so that the arc column region part of the welding process is separately represented by one of said model components in the electric circuit model.\",\n \"By such a method, together with a suitable physical model, important information is obtained such as arc mean temperature, temperature near to the wire, temperature near to the work piece, arc radius and arc current-voltage characteristics, and it is also possible to take the thermophysical properties of a shielding gas present in the arc column region into consideration in determining said at least one welding parameter.\",\n \"According to another preferred embodiment of the invention said methods comprise the step of: dividing the welding process so that the wire part of the welding process is separately represented by one of said model components in the electric circuit model.\",\n \"By such a method, together with a suitable physical model, important information is obtained such as metal transfer mode, dipping frequency etc, and it is also possible to take material properties of the wire into consideration in determining the value of said at least one welding parameter.\",\n \"According to another preferred embodiment of the invention said methods comprise the step of: dividing the welding process so that the work piece part of the welding process is separately represented by one of said model components in the electric circuit model.\",\n \"By such a method, together with a suitable physical model, important information is obtained about quality features such as weld joint profile, penetration profile etc, and it is also possible to take material properties of the work piece into consideration in determining the value of said at least one welding parameter.\",\n \"A fourth object of the invention is to provide a device which has an arrangement for controlling the arc welding equipment during a welding operation by adjusting at least one welding parameter determined without the need of measurements of the welding process or repeated welding experiments prior to welding.\",\n \"The fourth object is obtained according to the invention by providing such a device, referred to in the introduction, in which the means included in the control arrangement is adapted to use said theoretical model comprising at least two separate parts, of which at least one part represents the arc or a portion thereof, corresponding to different parts of the welding process, each model part being represented by a model component, said model components together with a model power source being included in an electric circuit model, and adapted to calculate at least one electric circuit model parameter related to said at least one welding parameter from the electric circuit model for said control by said member.\",\n \"Thus, by the means according to the invention it is possible to determine how to set at least one welding parameter value, such as the welding current or supply voltage, wire feed rate, wire extension etc., with the purpose of optimizing the welding process under the present conditions, and use this welding parameter value by said member for adjusting the arc welding equipment in accordance with the present conditions to obtain a weld with the properties desired.\",\n \"According to a preferred embodiment of the invention said means is adapted to use said theoretical model having the arc-wire interaction region part of the welding process separately represented by one of said model components in the electric circuit model.\",\n \"By such a device, together with a suitable physical model, important information is obtained because it is possible to take important factors in the welding process, such as the voltage drop over the region near to anode and the heat flux from arc to anode, into consideration in determining said at least one welding parameter value.\",\n \"In the arc-wire interaction region of the welding process, high gradients of different quantities such as temperature and concentration of particles are present, and the voltage drop in this region is very high.\",\n \"Thus, this region gives an important contribution to the characteristics of the welding process.\",\n \"According to another preferred embodiment of the invention said means is adapted to use said theoretical model having the arc-work piece interaction region part of the welding process separately represented by one of said model components in the electric circuit model.\",\n \"By such a device, together with a suitable physical model, important information is obtained because it is possible to take important factors in the welding process, such as the voltage drop over the region near to cathode and the heat flux from arc to cathode, into consideration in determining said at least one welding parameter value.\",\n \"In the arc-work piece interaction region of the welding process, high gradients of different quantities such as temperature and concentration of particles are present, and the voltage drop in this region is very high.\",\n \"Thus, this region gives an important contribution to the characteristics of the welding process.\",\n \"According to another preferred embodiment of the invention said means is adapted to use said theoretical model having the arc region part of the welding process separately represented by one of said model components in the electric circuit model.\",\n \"By such a device, and particularly together with an embodiment of the invention in which the means is adapted to use said theoretical model having information obtained through calculations from a physical model of the arc column region part of the welding process to be used in determining said model component representing the arc region part, important information is obtained such as arc mean temperature, temperature near to wire, temperature near to work piece, arc radius and arc current-voltage characteristics, and it is also possible to take the thermo-physical properties of a shielding gas present in the arc region into consideration in determining said at least one welding parameter.\",\n \"According to another preferred embodiment of the invention said means is adapted to use said theoretical model having information obtained through calculations from a physical model of the arc-wire interaction region part and/or of the arc-work piece interaction region part of the welding process to be used in determining said model component representing the arc region part, important information is obtained because it is possible to take important factors in the welding process, such as the voltage drop over the region near to anode/cathode and the heat flux from arc to anode/cathode, into consideration in determining said at least one welding parameter value.\",\n \"In the arc-wire interaction region and in the arc-work piece interaction region of the welding process, high gradients of different quantities such as temperature and concentration of particles are present, and the voltage drop in these regions is very high.\",\n \"Thus, these regions give an important contribution to the characteristics of the welding process.\",\n \"According to another preferred embodiment of the invention in which said means is adapted to use said model component determined by means of said physical model describing the arc by at least one equation independent of the extension in space of the arc, and particularly together with an embodiment of the invention in which said means is adapted to use said model component determined by means of said physical model describing the arc by said at least one equation only dependent on the time, the model component and thus said at least one welding parameter value, may be determined fast and with proportionally short computational time, which makes the device very suitable to be used in controlling the arc welding equipment on line.\",\n \"According to another preferred embodiment of the invention said means is adapted to use said theoretical model having the arc column region part of the welding process separately represented by one of said model components in the electric circuit model.\",\n \"By such a device, together with a suitable physical model, important information is obtained such as arc mean temperature, temperature near to wire, temperature near to work piece, arc radius and arc current-voltage characteristics, and it is also possible to take the thermo-physical properties of a shielding gas present in the arc column region into consideration in determining said at least one welding parameter.\",\n \"According to another preferred embodiment of the invention said means is adapted to use said theoretical model having the wire part of the welding process separately represented by one of said model components.\",\n \"By such a device, together with a suitable physical model, important information is obtained such as metal transfer mode, dipping frequency etc, and it is also possible to take material properties of the wire into consideration in determining the value of said at least one welding parameter.\",\n \"According to another preferred embodiment of the invention said means is adapted to use said theoretical model having the work piece part of the welding process separately represented by one of said model components.\",\n \"By such a device, together with a suitable physical model, important information is obtained about quality features such as weld joint profile, penetration profile etc, and it is also possible to take material properties of the work piece into consideration in determining the value of said at least one welding parameter.\",\n \"The invention also relates to computer program products according to the claims enclosed.\",\n \"It is obvious that the methods according to the invention defined in the sets of method claims enclosed are well suited to be performed through program instructions from a processor which may be effected by a computer program.\",\n \"Further advantages of and features of the methods and the device, the computer program products, and the uses of the methods appear from the following detailed description and the other claims enclosed herein.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS With reference to the appended drawings, below follows a description of preferred embodiments of the invention cited as examples.\",\n \"In the drawings: FIG.\",\n \"1 is a schematic illustration of an arc welding process, FIG.\",\n \"2 is a schematic illustration of the voltage distribution in the gap between a wire and a work piece in an arc welding operation, FIG.\",\n \"3 is an illustration of an electric circuit model representing an arc welding process, FIG.\",\n \"4 is a schematic illustration of a device according to the invention and of applications of the invention, FIG.\",\n \"5 is a block diagram of a method according to the invention, FIG.\",\n \"6 is a block diagram of another method according to the invention, FIG.\",\n \"7 is a block diagram of another method according to the invention, FIG.\",\n \"8 is a schematic illustration of a physical model of the arc, FIG.\",\n \"9 is an illustration of the geometry of an arc defined in a physical model, FIG.\",\n \"10 is an illustration of a physical model of the arc-wire interaction region in an arc welding process, FIG.\",\n \"11 is an illustration of a physical model of the arc-work piece interaction region in an arc welding process, FIG.\",\n \"12 is an illustration of the energy balance in the ionization zone, FIG.\",\n \"13 shows different kinds of welding parameters, FIG.\",\n \"14 is a schematic definition for work piece modelling of a horizontal-vertical fillet joint produced by gas metal arc welding, FIG.\",\n \"15 shows an example of a non-linear regression model, and FIGS.\",\n \"16 and 17 are analytic models of work piece.\",\n \"DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS OF THE INVENTION In this application the wording “welding process” is used as a generic term of the main parts included in an arc welding operation and of the physical processes associated therewith which take place when an electric arc is generated between a wire (anode) and a work piece (cathode).\",\n \"In FIG.\",\n \"1 an arc welding process is schematically illustrated.\",\n \"The power source 1 is connected to a wire 2 and a work piece 3 to create a closed electric circuit by means of an arc 4 generated in the gap between the wire 2 and the work piece 3 with the purpose of transferring material from the wire 2 to the work piece 3.The welding process may be divided in the following main parts: wire part 5, arc region part 6 and work piece part 7.Furthermore, the arc region part 6 may be divided in subparts, namely an arc-wire interaction region part 8, an arc column region part 9 and an arc-work piece interaction region part 10.All these parts contribute to the characteristics of the welding process and to a certain voltage drop in the electric circuit.\",\n \"However, in the electrode regions, i.e.\",\n \"in the arc-wire interaction region part 8 and the arc-work piece interaction region part 10 of the welding process, the voltage drop is very high compared to the voltage drop in the central part of the arc, i.e.\",\n \"in the arc column region part 9.In FIG.\",\n \"2 the voltage distribution along the arc, or in other words in the gap between the wire 2 and the work piece 3, is schematically illustrated.\",\n \"It is indicated that the voltage drops highly in the regions near to the wire and near to the work piece.\",\n \"This means that the regions situated near to the wire and near to the work piece are important parts in the welding process and have strong influence on the weld joint resulting from the welding operation.\",\n \"Furthermore, the voltage distribution in the gap is very difficult to determine by direct measuring of the arc whereas the voltage drops due to the resistances and inductances of the conduits 11 in the circuit and of the power source 1 itself are easily determined by conventional measurements.\",\n \"Thus, there is a need of a physical model for describing the phenomena of the arc to predict the behaviour of the welding process.\",\n \"However, as already indicated the wire 2 and work piece 3 strongly interact with the arc 4 and influence the welding process.\",\n \"Thus, good physical models of these parts are also needed to describe the welding process in its entirety.\",\n \"The main idea of the invention is to obtain the value of at least one welding parameter by means of a theoretical model and use said at least one welding parameter in operating an arc welding equipment and/or in simulating an arc welding process and/or in predicting the quality of a weld obtained from an arc welding operation.\",\n \"This is performed by dividing the welding process into parts in a theoretical model and letting each of these welding process parts and the welding parameters associated therewith be represented by a model component.\",\n \"The components are then put in an electric circuit model together with a model power source with the purpose of calculating at least one electric circuit model parameter related to said at least one welding parameter from the electric circuit model.\",\n \"The components may be resistive and/or inductive components, but also other electric elements than pure resistors and inductors may be included in the electric circuit model.\",\n \"Furthermore, the model components may be connected in series and/or in parallel.\",\n \"In FIG.\",\n \"3 an example of an electric circuit model 12 including model components 13 and a model power source 14 is illustrated.\",\n \"In this electric circuit model 12 the wire part 5 is represented by a resistor, Rw, the work piece part 7 by a resistor, Rwp, and the arc part 6 is represented by a resistor, Ra.\",\n \"In addition to these resistances it is necessary to define a model resistance R0 and a model inductance L0 corresponding to the resistance and the inductance from the power source 1 itself and the cables 11 in the real welding circuit.\",\n \"The circuit model resistance R0 and the inductance L0 are well known properties, or may be chosen, and for the simplification below these are treated as a single component (RL)rest.\",\n \"(In this component also other resistances, such as a contact resistance between the wire and a nozzle holding the wire, may be included).\",\n \"Although the electric circuit model 12 illustrated in FIG.\",\n \"3 comprises four separate components: Rw, Rwp, Ra and (RL)rest, in accordance with the invention the welding process may be divided into two or more parts, but at least two separate parts, of which at least one part represents the arc or a portion thereof present in the welding process.\",\n \"In the case of two separate model components 13 resistances in the circuit, with the exception of the resistance Ra from the arc region part 6, are suitably included in (RL)rest.\",\n \"Of course higher dissolution, i.e.\",\n \"more model components 13 representing different parts in the welding process, gives a more precise description of the arc welding process and also the possibility to obtain information about more welding parameters and to take more characteristics of the welding process into consideration.\",\n \"However, it is also possible to use different physical models representing subparts of a part of the welding process, the arc region part 6 for instance, which are based on the different characteristics of the part in different regions, for example the arc-wire interaction region 8, the arc column region 9 and/or the arc-work piece interaction region 10, resulting in one common model component such as a resistance Ra.\",\n \"In this way different parts of the welding process may be described without using a separate model component 13 for each part.\",\n \"In addition it is also possible to use input values from other parts of the welding process when the model component is calculated.\",\n \"By using Kirchoff's Voltage Laws at least one electric circuit model parameter may be calculated from the electric circuit model 12.For example, if the voltage supply 14 and the other components 13 are known, the current may be calculated or vice versa for a given current an unknown model component may be calculated.\",\n \"The model components Rw, Ra and Rwp in FIG.\",\n \"3 and other such components may be calculated from one or more of the physical models of the wire, arc and the work piece and of their interaction with each other.\",\n \"Below, examples of physical models are given.\",\n \"In FIG.\",\n \"4 the invention and some of its applications are schematically illustrated.\",\n \"The device according to the invention comprises an arc welding equipment 21 to be used in a welding operation for creating a weld joint.\",\n \"The welding equipment 21 are suitably an automated or robotic arc welding equipment which may be adjusted by varying at least one welding parameter, such as supply voltage, shielding gas flow rate, welding speed, torque angle, wire extension, wire feed rate etc.\",\n \"Furthermore, the device includes an arrangement 22 for controlling the operation of the arc welding equipment 21.The control arrangement 22 comprises a means 23, for example a processor and computer programs and electric components/circuits thereto, adapted to determine the value of said at least one welding parameter while using a theoretical model 24 of the welding process, and a member 25, for example any kind of control equipment which is able to communicate with the welding equipment and said means and thus transfer a signal representing the welding parameter value to the welding equipment, for adjusting the welding equipment 21.The adjustment may be performed prior to welding and/or on line during the welding.\",\n \"The means 23 is adapted to use said theoretical model 24, which comprises at least two separated parts 26, 27, of which at least one part represents the arc or portion thereof, corresponding to different parts of the welding process.\",\n \"Each model part 26, 27 is represented by a model component 13, see also FIG.\",\n \"3, and these model components 13 together with the model power source 14 are included in the electric circuit model 12.The means 23 is adapted to calculate at least one electric circuit model parameter 13 from the electric circuit model 12, and to use the value of this parameter related to said at least one welding parameter, for said control of the welding equipment 21 through adjusting of said at least one welding parameter of the welding process by said member 25.As indicated in FIG.\",\n \"4 the means 23 may be used without the member 25 and the arc welding equipment 21 for other purposes than direct controlling of an arc welding equipment.\",\n \"Thus, such a means may be used for simulation 28 of any arc welding process and/or for prediction of the quality 29 of a weld resulting from an arc welding operation.\",\n \"Furthermore, said means 23 and/or the control arrangement 22 may be used in performing any of the methods described herein.\",\n \"In FIG.\",\n \"5 a block diagram of the method for controlling an arc welding equipment used in a welding operation and adjustable by varying at least one welding parameter value is illustrated.\",\n \"The method comprises the steps of determining 30 said at least one welding parameter value while using 31 a theoretical model of the welding process associated with the welding operation, controlling 32 the operation of the welding equipment and the welding process associated therewith by using said at least one welding parameter value for adjusting the welding equipment, where the determining comprises the steps of dividing 33 the welding process into at least two separate parts, of which at least one part represents the arc or a portion thereof, in the theoretical model, representing 34 each of said at least two welding process parts and the welding parameter/parameters associated therewith by a model component, putting 35 the model components and the model power source in an electric circuit model, and calculating 36 at least one electric circuit model parameter related to said at least one welding parameter from the electric circuit model.\",\n \"In FIG.\",\n \"6 a block diagram of the method for simulating an arc welding process is illustrated.\",\n \"The method comprises the steps of inputting 40 data into a theoretical model representing the welding process, determining 41 at least one welding parameter value of the welding process while using 42 the theoretical model and the data input, with the purpose to simulate 43 the welding process, where the determining comprises the steps of dividing 44 the welding process into at least two separate parts, of which at least one part represents the arc or a portion thereof, in the theoretical model, representing 45 each of said at least two welding process parts and the welding parameter/parameters associated therewith by a model component, putting 46 the model components and a model power source in an electric circuit model, and calculating 47 at least one electric circuit model parameter related to said at least one welding parameter from the electric circuit model.\",\n \"In FIG.\",\n \"7 a block diagram of the method for predicting the quality of a weld obtained from an arc welding operation is illustrated.\",\n \"The method comprises the steps of inputting 50 data into a theoretical model representing the welding process associated with the welding operation, determining 51 at least one property of the weld related to at least one welding parameter value of the welding process while using 52 the theoretical model and the data input, where the determining comprises the steps of dividing 53 the welding process into at least two separate parts, of which at least one part represents the arc or a portion thereof, in the theoretical model, representing 54 each of said at least two welding process parts and the welding parameter/parameters associated therewith be represented by a model component, putting 55 the model components and the model power source in an electric circuit model, and calculating 56 at least one electric circuit model parameter related to said at least one welding parameter, and related to said at least one property of the weld, from the electric circuit model.\",\n \"The methods may be used for different objects such as for predicting the metal droplet formation in an arc welding operation for a given set of welding parameters, for predicting the metal transfer mode in an arc welding operation for a given set of welding parameters, for calculating at least one welding parameter value required for obtaining a particular metal transfer mode in a welding operation and/or for calculating at least one default setting value to be used for adjusting an automatic arc welding equipment prior to welding.\",\n \"The prediction of quality may be used for example for prediction of any mechanical property of a weld joint for a given input, i.e.\",\n \"set of welding parameters, or in the reverse direction for determining one or more welding parameter values for a given input, i.e.\",\n \"any mechanical property of a weld joint.\",\n \"An important application of the methods and the device according to the invention is the prediction of the characteristics of a certain welding process, and as mentioned above the prediction of the metal transfer mode present during an arc welding operation, which makes it possible to predict the transfer mode as a function of a certain welding parameter, such as wire feed speed or current, and thus to predict the transition region between so called short arc and spray arc mode.\",\n \"In different embodiments of the methods the step of dividing the welding process may include dividing of the welding process so that the arc-wire interaction region part and/or the arc region part and/or the arc column region part and/or the wire part and/or the work piece part is separately represented by one of said model components 13 in the electric circuit model 12.In embodiments of the methods using an electric circuit model 12 according to FIG.\",\n \"3 said methods comprise the steps of dividing the welding process so that the arc region part 6, the wire part 5, the work piece part 7 (and the part corresponding to (RL)rest) each is separately represented by one of said model components 13 in the electric circuit model 12.Furthermore, in a preferred embodiment described more in detail below, said model component, Ra, representing the arc region part is determined by means of information obtained through calculation from physical models of the arc column region part 9, the arc-wire interaction region part 8, and the arc-work piece interaction region part 10.In FIGS.\",\n \"8, 9, 10 and 11 an example of a physical model, including submodels, based on energy, mass and momentum equations is schematically illustrated.\",\n \"As indicated in FIG.\",\n \"8 the wire, the arc, the work piece, the arc-wire interaction and the arc-work piece interaction are taken into account.\",\n \"In FIG.\",\n \"9 the arc column 60 is assumed to be a cylindric conductor with a mean diameter 61 and an arc expansion transition zone 62 near to the wire 63.In FIG.\",\n \"10 the arc-wire interaction region part of the welding process is divided into a first zone 64 nearest to the wire, called the space charge zone, and a second zone 65 nearest to the arc column, called the ionization zone, for describing the properties of the zones through different physical submodels.\",\n \"Also the arc-work piece interaction region part of the welding process in FIG.\",\n \"11 is divided into a first zone 66 nearest to the work piece, called space charge zone, and a second zone 67 nearest to the arc column, called the ionization zone, for describing the properties of the zones through different physical submodels.\",\n \"Below follows a more detailed description of possible physical models which may be used in connection with the invention.\",\n \"PHYSICAL MODELS Arc Model As already mentioned the welding process may be divided into different parts and the arc region part may be divided in subparts, namely an arc-wire interaction region part, an arc column region part and an arc-work piece interaction region part.\",\n \"The arc column, described in this chapter, occupies most of the space between the wire and the work piece.\",\n \"In this region, gradients of thermodynamic variables, such as temperature and pressure, are not too high and the system is in local thermodynamic equilibrium.\",\n \"The arc model suggested here is based on energy, mass and momentum equations.\",\n \"The arc current generates Lorentz forces which have an important role in the welding process.\",\n \"The arc is usually constricted towards the wire tip.\",\n \"This rises the axial component of the Lorentz force.\",\n \"Plasma is therefore accelerated towards the work piece.\",\n \"The electromagnetic forces in the welding process are important both for the stabilization of the arc and for material transfer to the work piece.\",\n \"To be able to predict some phenomena of welding arcs, thermo-physical properties of shielding gas in the temperature range 300-30000 Kelvin are required.\",\n \"Due to arc-wire and arc-work piece interactions, the shielding gas is usually contaminated by metallic vapours, which strongly influence the physical properties of the arc.\",\n \"The structure for the arc model proposed herein is shown in FIG.\",\n \"8.This structure gives the possibility to consider the effects of the shielding gas, the wire material and the work piece material.\",\n \"According to the state of art, a detailed two-dimensional simulation of the arc is known today.\",\n \"However, such a simulation requires long computational time.\",\n \"This makes it difficult to use such a tool for controlling and monitoring the arc welding process.\",\n \"In this application instead a fast arc model with the structure illustrated in FIG.\",\n \"8 and 9 is proposed.\",\n \"The model is based upon the following considerations: the arc is in local thermodynamic equilibrium, the arc is assumed to be a cylindrical conductor with a uniform temperature and a mean diameter, an arc expansion transition zone between the wire and the arc column is considered, the arc mean temperature and diameter are obtained from the arc energy balance and the Steenbeck minimum principle.\",\n \"This principle is based on the minimization of the arc power loss, thermo-physical properties are calculated in separate modules and coupled to the arc model, and the arc-wire interaction and the arc-work piece interaction are taken into account.\",\n \"The energy balance of the arc is given below: LI 2 σ A ︸ dissipation in arc column + ( V an + V ca ) I ︸ dissipation due to anode and cathode falls = A ρ v ( h - h 0 ) ︸ convection + ALU rod ︸ radiation + Q A - A ︸ Heat transfer Arc - Anode + Q A - C ︸ Heat transfer Arc - Cathode where L is the arc length, I denotes the electric current, σ refers to the electric conductivity, A is the cross section area of the arc column, Van and Vca are the voltage drops in the near to anode (wire) region and in the near to cathode (work piece) region, respectively.\",\n \"ρ is the gas density, h and h0 are the specific enthalpies of the arc and the surroundings, and Urad is the mean radiation density, which all are functions of the arc temperature.\",\n \"ν denotes the mean gas velocity.\",\n \"The heat transfer terms for arc-anode QA-A and arc-cathode QA-C are obtained from detailed physical models of the arc-cathode and arc-anode interactions.\",\n \"Using Ampere's law and integration of Navier-Stokes equations for an incompressible flow in the expansion zone a simple equation for the mean velocity can be deduced as v = C a 1 R Arc π C b μ o 2 ρ ( R Arc 2 R Wire 2 - 1 ) where Rarc and Rwire are the radii of the arc column and wire, respectively, and μ0 is the magnetic field constant.\",\n \"Constants Ca and Cb depend on the current density profiles on the anode wire and will be derived with the help of CFD simulations and simple experiments.\",\n \"The equations above together with the minimization of the power loss provide all necessary information such as arc mean temperature, arc radius and arc current-voltage characteristics.\",\n \"The benefits of the proposed arc model are: the arc model may be executed very fastly and thus it may be used as a part of a software for performing the invention and for example be used together with a model for the arc-wire interaction so that droplet formation and transport may be predicted.\",\n \"Arc-work Piece Interaction Model Near the electrodes, high gradients of temperature and concentration of particles are present.\",\n \"These lead to deviations from thermal and chemical equilibrium.\",\n \"It is well known that the voltage.\",\n \"drops in the near to wire region and in the near to work piece region are very high and dependent on electrode materials and arc column plasma properties.\",\n \"Therefore, it is important to take physical properties of the near electrodes phenomena for prediction of an arc welding process into consideration.\",\n \"In this section the model for the interaction between the arc and the work piece is presented.\",\n \"The model is based on the following: the region near the work piece is divided into two zones: the ionization zone and the space charge zone, collisions between particles are negligible in the space charge zone, electrons and heavy particles are considered as two fluids with two different temperatures, densities of ions and electrons are equal in the ionization zone, densities of ions and electrons are not equal in the space charge zone, the electron emission mechanism is thermo-field emission, and equations describing the two zones are integrated over the zones.\",\n \"The inputs to the arc-work piece interaction region model are the work piece material and the plasma properties in the arc column region (e.g.\",\n \"temperature and pressure).\",\n \"The outputs of the model are the voltage drop over the arc-work piece interaction region and the heat flux from work piece to arc due to charged particles.\",\n \"Furthermore, there is a conductive heat flux due to neutral particles.\",\n \"These data are needed in the global energy balance of the arc column.\",\n \"The model is adapted in a way so that it may easily be coupled to the arc model described above.\",\n \"The benefits of the proposed arc-work piece interaction model are: the model may be coupled to the arc column model and/or to the arc-wire interaction region model and allows a more complete description of the arc, the model gives accurate predictions of the heat flux into the work piece which is an important factor that effects the quality of the weld, the model predicts the rate of metal vapours from the work piece, the model may be executed very fastly and thus it is for instance not necessary to decrease the speed of a welding operation when using the model for controlling an arc welding operation, and the model considers explicitly the properties of the work piece material, for example the work function and the thermal conductivity.\",\n \"For illustrating the influence of the work piece material on the arc welding process, a summary of the physics is presented in the following.\",\n \"The energy flux from the plasma to the work piece is given by: q = J i [ k ( T i 2 + T e 2 - T w 2 ) + e ( φ c + E i - ϕ eff ) ] + J e ( T c 2 k + ϕ eff e ) - J em ( T w 2 k + ϕ eff e ) - q vap where Je, Jem and Ji are the plasma back-dffused electrons current density, the emitted electrons current density and the ion current density, respectively.\",\n \"Tw, Te and Ti indicate the temperature of the cathode wall, electrons and ions.\",\n \"φc and Ei are the voltage drop in the space charge zone and the ionization energy of gas.\",\n \"φeff is the effective work function, which is defined as the difference between the mean energy of electrons inside the metal and the mean energy of the emitted electrons just outside the metal.\",\n \"qvap is the energy flux carried away by vaporization.\",\n \"The energy balance in the ionization zone may be summarized as shown in FIG.\",\n \"12.This figure shows that a part of the total power density, the Joule effect, received by the electron gas is transferred to the heavy particles by elastic and inelastic collisions.\",\n \"The other part of energy is lost by radiation and by thermal flow to the cold walls.\",\n \"According to earlier investigations of the ionization zone, the main losses of electronic energy is due to ionization.\",\n \"Based on these results, the energy losses due to elastic collisions can be neglected.\",\n \"The energy balance is simplified by integration over the zone: Jem(2kTw+eφc)+Joule_effect=Je(2kTe+eφc)+eJiE+3.2/kTe The first term on the left hand side is the flux of the electrons brought into the ionization zone by the emitted electrons accelerated in the space charge zone.\",\n \"The second term of the left hand side presents the work of the electric field over the electrons inside the ionization zone.\",\n \"The first term on the right hand side is the energy flux carried away by the electrons leaving the ionization for the space charge zone.\",\n \"eJiE is the losses of the electron energy due to ionization.\",\n \"Finally, the last term on the right hand side is the energy flux carried away by electrons leaving the ionization zone for the arc column.\",\n \"To be able to predict the cathode region behaviour, the current densities Je, Jem and Ji are required.\",\n \"These current densities are obtained by detailed kinetics of particles in the space charge zone.\",\n \"Here only the results are given.\",\n \"The ion current density and the back-diffused electron current density are given by J i = n io e k ( T i + T e ) m J e = n io e 4 8 k T e π m e exp ( - φ c e T e k ) According to the thermionic field emission mechanism, the emitted electron current density is given by Richardson-Dushman equation with the Schottky correction.\",\n \"J em = k 2 4 π m e e h 3 T w 2 exp ( ϕ eff e T w k ) In the above equation, h and me are the Planck's constant and the electron mass, respectively.\",\n \"The effective work function is connected to the electric field according to: ϕ eff = ϕ 0 - ( E r e 4 πɛ ) where Φ0, ∈ and EC are the work function of the work piece material, the dielectric constant and the electric field strength at the cathode wall.\",\n \"The electron and ion densities at the edge of the space charge zone can be calculated with good approximation as: n i0 = n r0 = n plasma 0.8 2 + α where α = ( T i k m i D io k r n eq 2 ) 1 2 Di0 and kr are the ion-neutral diffusion coefficient at the top of the equilibrium limit and the recombination rate.\",\n \"The set of equation presented is closed and can be solved easily to predict cathode region properties or arc-work piece interaction region properties in the arc welding process.\",\n \"Arc-wire Interaction Region Model In this section the model for the interaction between the arc and the wire (anode region) is presented.\",\n \"The model is based on the following: the region near the wire is divided into two zones: the ionization zone and the space charge zone, collisions between particles are negligible in the space charge zone, electrons and heavy particles are considered as two fluids with two different temperatures, densities of ions is much less than densities of electrons in space charge zone, temperature of electrons remains constant through anode region, the voltage drop in the ionization zone is negligible, and no electron emission from the wire is assumed.\",\n \"The inputs to the arc-wire interaction region model are the wire material and the plasma properties in the arc column region (e.g.\",\n \"temperature and pressure).\",\n \"The outputs of the model are the voltage drop over the arc-wire interaction region and the heat flux from arc to wire due to charged particles.\",\n \"These data are needed in the global energy balance of the arc column.\",\n \"The model is adapted in a way so that it may easily be coupled to the arc model described above.\",\n \"The benefits of the proposed arc-wire interaction model are: the model may be coupled to the arc column model and/or the arc-work piece interaction region model and allows a more complete description of the arc, the model gives accurate predictions of the heat flux into the wire which is an important factor for describing the welding process, the model may be executed very fastly and thus it is for instance not necessary to decrease the speed of a welding operation when using the model for controlling an arc welding operation, and the model considers explicitly the properties of the wire material, for example the work function.\",\n \"For illustrating the influence of the wire on the arc welding process, a summary of the physics is presented in the following.\",\n \"The total energy flux from the plasma to the wire is given by the losses from the arc due to charged particles, thermal conduction and radiation minus the heat power utilized for evaporation and radiation of the wire tip surface Q .\",\n \"= Q .\",\n \"el + Q .\",\n \"cond + Q .\",\n \"rad ︸ energy flux from arc - Q .\",\n \"gy , wire - Q .\",\n \"rad , wire ︸ energy flux from wire tip surface For the energy flux of charged particles, electrons are considered only, since the ion current density flux in the arc-wire interaction region is low compared to the electron current density flux Q .\",\n \"el = j e ( 5 k b 2 e T e + V an + ϕ ) There is energy flux due to thermal energy of electrons, kinetic energy of electrons resulting from acceleration in electric field and potential energy of electrons after phase transformation on the wire surface (free to bound).\",\n \"je is the current density of the electrons and Te is the electron temperature in the arc-wire interaction region.\",\n \"The electron temperature is taken as the mean arc core temperature provided by the arc model.\",\n \"φ is the nominal work function of the wire surface (known), and Van is the voltage drop over the anode region, which consists mainly of the voltage drop in the space charge zone Van=φan where je is obtained from kinetics je=je(ne,φan) where ne is the electron density in the anode region and is calculated using the equation of state, thermodynamic equilibrium laws and the thermodynamic properties of the arc (temperature, pressure).\",\n \"For obtaining the voltage drop over the arc-wire interaction region the ion current density ji is taken into account.\",\n \"The following relation can be derived from kinetics ji=ji(ni,φan) where ni is the ion density in the ionization zone, which is equivalent to the electron density ne.\",\n \"Knowing the total current density j j=je+ji the voltage drop over the arc-wire interaction region can be obtained.\",\n \"Thermal conduction from the arc results from the temperature gradient in the anode region acting on ions, and the heat fluxes due to the kinetic energy of ions and electrons in the arc-wire interaction region Q .\",\n \"cond = λ ∂ T ∂ n + n i v i h i + n e v e h e where vi and ve are the velocities of ions and electrons, respectively, which are given by kinetics at thermodynamic equilibrium.\",\n \"The radiation power from the arc is the fraction of the power totally radiated by the arc that reaches the wire tip surface {dot over (Q)}rad=∈effσTarc4 where ∈eff is an effective emission coefficient of the arc, which can be estimated from the emission coefficient of the arc column, assuming a cylinder, times the fraction of cylinder surface directed towards the wire.\",\n \"The energy flux due to evaporation of the wire tip surface is given by the product of evaporation rate and latent heat of the wire material {dot over (Q)}ev,wire={dot over (m)}Δh where the evaporation rates may be expressed as a function of arc column pressure and wire tip surface temperature {dot over (m)}={dot over (m)}(P,Tans) The energy flux due to radiation from the wire tip surface is given by {dot over (Q)}rad,wire=∈σTans4 The only unknown remaining is the wire tip surface temperature Tans.\",\n \"This temperature can be obtained from the wire model.\",\n \"A simple estimation is that it is equal to the melting temperature of the wire material.\",\n \"The set of equations presented above is closed and can be solved to predict the anode region properties or arc-wire interaction region properties in the arc welding process.\",\n \"Wire Model In the arc welding process, heat transfer from the arc to the wire causes rapid formation of droplets at the wire tip.\",\n \"Impact of detached and transferred droplets through the arc to the work piece may result in volume deformations and alteration of the mechanical properties of the weld.\",\n \"In the following, a model is suggested for prediction of the mass, heat and momentum transfer from dripping droplets into the work piece.\",\n \"The shape of melt metal at wire tip is influenced by time-dependent forces such as gravity, inertia, surface tension, electromagnetic force, the effect of arc pressure, viscous drag by the gas flow.\",\n \"An extensive description of thermophysical phenomena involved in mechanisms of formation of droplets may be formulated accurately with 2-D computer tools based on a detailed analysis of the plasma-electrode interactions in conditions of local thermodynamic equilibrium of the plasma and laminar flow in the calculation domain.\",\n \"However, calculations required for these operations may be time-consuming and not acceptable in a user interface software to be used for controlling and/or monitoring the quality of a weld in an welding operation.\",\n \"A simplified approach to the problem can be to consider the one-dimensional energy balance of a wire dominated by the effects of evaporation, metal transfer from the wire, ohmic heating in the wire by the arc current and current transfer from the plasma.\",\n \"In a quasisteady state mode, the total energy balance of the wire can be written as: Ev+Em=Eoh+Ec when the heat transfer processes for integrated values over the total volume of the wire stick-out are defined as: E v = ∫ S q ev L ⅆ S due to the evaporation from the surface to the tip of the wire; E m = ρ v 0 Sc ∫ T = 300 T mean C p ⅆ T the heat losses due to metal transfer in the form of droplets detaching from the wire tips; E oh = ∫ V w j 2 σ ⅆ υ the heating in the wire by the arc current, and E e = ∫ S [ j i ( V i - V s + 5 k b 2 e T ) + j e ( ϕ + 5 k b 2 e ) T p ] ⅆ S the heat transferred from the plasma to the droplet at the tip of the wire.\",\n \"In the above relations, Vw is the total volume of the wire stick-out, S is the total external surface of wire, and Sc=πr2 is the cross-sectional area of the wire at the end of the wire at z=0 (T=300 K) and r is its radius.\",\n \"The quantities Cp, κ and ρ are the specific heat, the thermal conductivity and density of the wire material at the local wire temperature, respectively.\",\n \"Tmean is the average droplet temperature at the time of detachment, and v0 is the wire feed speed.\",\n \"The evaporation rate in g.s−1 at the wire tip is denoted as qev=Pevma/4(makbTa/3)0.5 where Pev and ma are the vapour pressure and theatomic mass of the wire, respectively, while kb is the Boltzmann constant and Ta the temperature of the wire surface.\",\n \"Other symbols are the following: L, the latent heat of evaporation of the wire material, e, the elementary charge, ji the ion current density from the plasma, je the electron current density from the plasma; the total current density is denoted j; Tp is the temperature immediately in front of the wire, φ is the nominal work function of the wire surface; σ represents the electrical conductivity, Vi is the first ionization potential of the plasma gas and Vs the voltage drop across the sheath in the wire region.\",\n \"In this approximation, conventional heat conduction from the plasma through neutral particles, conduction through the end of the wire and black body radiation are neglected since their effect can theoretically be shown to be small compared to Ev, Em, Eoh and Ee.\",\n \"Heat transferred from the plasma to the wire may be expressed S a = j i ( V i - V s + 5 k b 2 e T a ) + j e ( ϕ + 5 k b 2 e T p ) - q ev L By solving the time-dependent energy conservation equation at small time steps, ρ ∂ h ∂ t + ρ v 0 ∂ h ∂ z = ∂ ∂ z ( κ ∂ T ∂ z ) + 5 k b 2 e ( j z C p ∂ h ∂ z ) + j 2 σ + F where F is a term referring to the heat input Sa, from the plasma, the temperature profile of the wire can be obtained, and therefore the molten length at the wire tip can be estimated as a function of time.\",\n \"Metal transfer in the form of a droplet will occur when the molten wire reaches a critical length Lcrit for a given set of welding parameters.\",\n \"Estimation of the volume of droplet can be deduced from a dynamic treatment in the wire region using simplified expressions of surface tensions, electromagnetic forces, gravity, arc pressure and viscous drag force around the molten metal at the tip of the wire at each time step.\",\n \"The initial shape of the electrode at a time t=0 can be taken as cylindrical with diameter equal to that of the wire covered by about a 0.1 cm thick layer of molten liquid at the tip of the wire.\",\n \"Initial conditions are to correspond to an arc operating with continuously moving wire of undeformed cylindrical shape.\",\n \"The inputs required for the wire model are the arc current, the wire feed speed, the wire diameter, the initial interelectrode separation distance, the torch angle and the characteristics of the arc.\",\n \"The latter may be supplied into the model by outputs from the arc model.\",\n \"The outputs include the detachment frequency, the volume of a detached droplet of molten metal, the average temperature in a droplet and the acceleration supported by the droplet in the axis of the arc column at detachment.\",\n \"This result will provide information for evaluation of reinforcement of joint for a given gas arc welding process and work piece geometry.\",\n \"Also available from the model will be the wire stick-out length and resistance, which allow a more accurate calculation of the arc length than from the simplified model described below.\",\n \"As a first step a simplified model for the wire has been used.\",\n \"The arc length is obtained from a balance between wire feed rate wfr and wire melting rate ⅆ ⅆ t ( L + τ ⅆ L ⅆ t ) = - w fr + I 2 R an + I δ V δ H where τ is a time constant that delays the change of arc length from change of heat input.\",\n \"The resistance of the wire stickout Ran is calculated assuming a constant stickout temperature R an = ζ A w ( D - L ) where ζ is the electrical resistivity of the wire, Aw the wire cross-section area, D the distance between the nozzle and the work piece.\",\n \"L is the arc length, which is the output of this model.\",\n \"The power input considers both Ohmic heating of the wire stickout and energy transfer from arc to wire.\",\n \"The wire melting rate is obtained by relating the power input to the wire enthalpy per unit length δH.\",\n \"The latter accounts for energy release due to electron absorption in anode and thermal energy carried by electrons δ V = ϕ an + 5 k B T e 2 e where φan is the work function of the anode material, kB is the Boltzmann's constant, Te is the electron temperature and e is the electron charge.\",\n \"Work Piece Model As part of multivariable processes, automated arc welding may be defined by so called indirect welding parameters (IWPs), acting upon a set of material parameters (MPs) with a resulting set of direct welding parameters (DWPs).\",\n \"IWPs should be chosen appropriately for given MPs.\",\n \"DWPs are to be explicitly fixed by specifications of e.g.\",\n \"optimal weld bead width, penetration and appearance.\",\n \"A basic issue to be considered regarding control for arc welding is to determine what is to be controlled and which parameters are accessible to control actions in the process.\",\n \"See FIGS.\",\n \"13 and 14.Quality control functionalities to be implemented in the proposed software could consist of: process stability monitoring, i.e.\",\n \"high speed tracking by e.g.\",\n \"timedomain statistical analysis based upon measured and predicted average IWPs, for example arc voltage or current intensity.\",\n \"It may allow detection of disturbing inputs (anomalies in gun-to-work piece distance, insufficient shielding gas coverage, etc.).\",\n \"Advantageously, vision sensors and sound probes also may be used, quality assurance of geometry and mechanical properties of weld joints (DWPS) by process parameters setting from fast advanced physical models based on MPs and IWPs.\",\n \"The models may include detailed arc-work piece interactions from weld pool phenomena (arc efficiency, droplet dripping, gas and impurities diffusion, etc.)\",\n \"and micro structural criteria (micro phases, residual stress, dislocations, etc.).\",\n \"One work piece model is illustrated in FIG.\",\n \"17.In practice, modelling of weld properties is often based on neural networks trained for a particular geometry of joints.\",\n \"Neural networks are parameterised non-linear regression models, see FIG.\",\n \"14.They can provide adapted solutions for treatment of intricate multi-variable systems when physical models are difficult to establish or do not exist.\",\n \"Also a program using neural networks should include mass, heat, and momentum transfers from the arc and dripping of droplets into work piece.\",\n \"Indeed, these quantities may have considerable consequences on the overall geometry of the weld and its intrinsic quality.\",\n \"An important component in the elaboration of a neural network is the control of the characteristic magnitudes of the weights (noted wj(i) in FIG.\",\n \"15) of input parameters.\",\n \"This operation requires that the network is preliminary trained from a set of known outputs when the weights are systematically changed until a best-fit description of the network is obtained as a function of the inputs.\",\n \"Among other possibilities for modelling of the work piece, a neural network program may be trained on gravity joints obtained under specified conditions (mild steel, gas shielding, wire speed, wire feed rate, plate thickness, etc) by an arc welding robot.\",\n \"Solidification of melt during GMA welding process depends on heat transfer conditions in the weld pool, see FIG.\",\n \"16.Fusion mechanism and characteristics of resultant welds are complex to model analytically.\",\n \"Neural networks can represent a suitable alternative for a model of a workpiece.\",\n \"Shape and mechanical properties of welds can strongly be affected by electromagnetic forces, buoyancy forces and surface tension forces that induce flow of molten metal in the weld pool possibly affected by impacts of droplets, vaporization, arc pressure and molecular contamination.\",\n \"Accurate enough predictions of weld quality by analytical model of those phenomena can be difficult to provide as they call for detailed computation and optimized calculation routines including appropriate numerical methods for moving boundary systems.\",\n \"Neural networks are powerful parameterised non-linear regression models for which no fundamental knowledge of the system to be modelled is required.\",\n \"They can be generally described as series of weighted variables (input parameters xi) yj=Σiwi(1)xi+θ(1) to form arguments of non-linear regression transfer functions (hidden variables).\",\n \"Example: hj=tanh yi Since a one hidden-unit model (hj=h, y=W(2)H+θ(2) output parameter) may not be sufficiently flexible, further degrees of non-linearity can be introduced by combining several non-linear regression transfer functions (output parameter y) yj=Σjwj(2)hj+θ(2) hence permitting neural networks to capture almost arbitrarily non-linear relationships.\",\n \"The capability of neural networks to handle complexity is much related to the number of regression transfer functions, i.e.\",\n \"the number of hidden layers.\",\n \"An important component in the elaboration of a neural network is the control of the characteristic magnitudes of the weights (wi(j)) of input quantities.\",\n \"This operation requires that the network is preliminary trained from a set of known outputs when the weights are systematically changed until a best-fit description of the network is obtained as a function of the inputs.\",\n \"Numerical procedures can be developed to avoid overfitting difficulties.\",\n \"In some cases, error estimates can also be calculated to define fitting uncertainty by considering probability distributions of sets of weights instead of using a unique one.\",\n \"A neural network may be produced to model effects of heat and mass transfers on weld geometry as part of an Arc Welding End-user Software.\",\n \"However, as already mentioned, many experimental parameters can influence an arc welding process (for example wire properties, shielding gas properties, power supply, wire feed speed, welding speed, electrode extension, etc.—see figure).\",\n \"A systematic investigation of merely 10 parameters at only three levels of variation would result in 310, i.e.\",\n \"about 59.103 possible combinations to explore.\",\n \"Theoretical knowledge of the arc suggests that the arc current, the welding speed and material properties are mainly responsible for heat and mass transfer into the workpiece.\",\n \"A GMAW process can be decomposed into separate interdependent modules for, respectively, the electrical circuit, the wire (anode), the arc, for the interactions between the arc and the wire (i.e.\",\n \"droplet formation), the interactions between the arc and the workpiece, and the workpiece itself (cathode).\",\n \"The amount of welds to be produced for making a neural network model of the workpiece can hence be reduced to a reasonably small number by successively comparing model results for the diverse modules to a few relevant experimental test cases characteristic of the functionality of the module tested, all critical parameters of other modules being fixed in other respects.\",\n \"In this aim, intern resistances, inductances and resistances on the welding equipment to be used for experiments should be measured beforehand and recording of process signals during welding (e.g.\",\n \"arc current and voltage, encoding signal form wire feeding system, etc.)\",\n \"should be carried out with an acquisition unit.\",\n \"Validation of module for electrical circuit can be achieved by comparing measured arc current and voltage with predicted values of these quantities at different wire feed speeds and supply voltages for different welding modes (short arc, spray, rapid-arc), all other parameters being kept constant and identical between models and experiments in other respects.\",\n \"Validation of the wire model for simulation of mass transfer mode (i.e.\",\n \"droplet dripping) can be achieved in a similar way by further measuring the average mass transfer rate from for several dimensions of wire diameter.\",\n \"Validation of the arc model can be achieved by varying supply voltage and electrode extension for different shielding gases and wire diameters, all other parameters being kept constant.\",\n \"Adjustments of each model may have to be implemented stepwise during validation of each module.\",\n \"Eventually, experiments to be carried out for modelling of complete effects of mass and heat transfer into workpiece simply concern the influence of torch and push/drag angles, welding speed, plate thickness, plate material and joint configuration, all other aspects being supposedly encompassed in the previously validated other routines.\",\n \"For maximum applicability of the software, experiments shall be carried out on different joints (for example overlap, T-joint) made e.g.\",\n \"in horizontal-vertical configuration of gravity configuration or other, depending on targeted market for the software.\",\n \"As input from the arc-dependent models to a neural network model of the workpiece, temperature distributions of electrons and ions, current intensity due to both species and evaporation rate at the workpiece surface will be accessible from a pre-calculated matrix established with the validated models.\",\n \"Mass, heat and speed of transferring droplets will also be referenced as inputs of the neural network from predictions obtained with the validated model for wire.\",\n \"The output of the neural network should enable a determination of the weld quality as defined either by geometry or mechanical properties.\",\n \"A complete version of the software is to include all possible aspects of quality.\",\n \"When quality is associated to weld shape, the outputs will consist of geometrical dimensions of welds.\",\n \"A sketch of predicted weld geometry will be displayed or dimensions according to the standard EN-25817 will be listed on end-user interface.\",\n \"When quality is associated to mechanical properties, the outputs will consist of predicted values of e.g.\",\n \"yield strength, ultimate tensile strength, Charpy toughness and temper embrittlement.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473819"}}},{"rowIdx":756,"cells":{"text":{"kind":"list like","value":[["Trusted authorization device","A trusted display (18) of a trusted authorization device (TAD) (10) displays on a trusted display (18) first information about a transaction to be authorized by a user (14) using a trusted keypad (20).","The TAD (10) generates (208) a random number (R); generates (1210) second information from the first information, the random number (R) and a first identification code (TADID-A) of the TAD (10); generates (212) a signature of the second information using a first encryption process; egnerates (216) a set of session keys (Ks1, Ks2, Ks3) by a second encryption process responsive to the random number (R) and a set of stored working keys (Kw1, Kw2, Kw3); and generates (218) third information by encrypting the second information and the signature using a third encryption process responsive to the set of session keys (Ks1, Ks2, Ks3).","A dat structure (42) is formed comprising the random numer (R), the first identification code (TADID-A), and the third information; and communicated (220) from the TAD (10) to the client (12) to a host server (28) for verification by a verification decryption server (32)."],["1.A method of providing for a trusted authorization of a transaction, comprising: a. providing for communicating with a first computer; b. providing for displaying first information to be authorized on a trusted display of a trusted authorization device, wherein said first information to be authorized is provided by said first computer; c. providing for receiving an authorization command from a trusted keypad of said trusted authorization device, wherein said authorization command is related to said first information; and d. if said authorization command provides for authorizing said first information, then providing for a set of operations by a trusted processor of said trusted authorization device, said set of operations comprising: i. generating a random number; ii.","generating second information that is responsive to said first information to be authorized, wherein said second information further incorporates both said random number and a first identification code associated with said trusted authorization device, wherein said first identification code is stored on a trusted memory of said trusted authorization device; iii.","generating a signature of said second information, wherein said signature is generated by a first encryption process; iv.","generating a set of session keys by a second encryption process, wherein said second encryption process is responsive to said random number and to a set of stored working keys, and said set of stored working keys are stored on said trusted memory of said trusted authorization device; v. generating third information by encrypting said second information and said signature using a third encryption process that is responsive to said set of session keys; and vi.","communicating to said first computer said random number, said first identification code, and said third information, wherein said random number and said first identification code are communicated in plaintext.","2.A method of providing for a trusted authorization of a transaction as recited in claim 1, further comprising providing for receiving a personal identification code from a user, wherein said personal identification code is incorporated in said second information.","3.A method of providing for a trusted authorization of a transaction as recited in claim 2, wherein said personal information code comprises a code entered with a trusted keypad of said trusted authorization device.","4.A method of providing for a trusted authorization of a transaction as recited in claim 2, wherein said personal information code is responsive to a biometric input from a user entered with a trusted biometric input device.","5.A method of providing for a trusted authorization of a transaction as recited in claim 2, wherein said personal information code is responsive to a signature input from a user entered with a trusted signature input device.","6.A method of providing for a trusted authorization of a transaction as recited in claim 2, wherein said personal information code is responsive to a voice input from a user entered with a trusted microphone.","7.A method of providing for a trusted authorization of a transaction as recited in claim 2, wherein said personal information code comprises a digest of at least one of a biometric input from a user entered with a trusted biometric input device, a signature input from a user entered with a trusted signature input device, and a voice input from a user entered with a trusted microphone.","8.A method of providing for a trusted authorization of a transaction as recited in claim 1, further comprising providing for receiving a personal identification code from a user, wherein the operation of providing for said set of operations by a trusted processor of said trusted authorization device is responsive to whether said personal identification code corresponds to a stored personal identification code, said stored personal identification code is stored in said trusted memory of said trusted authorization device, and said stored personal identification code is associated with an authentic user of said trusted authorization device.","9.A method of providing for a trusted authorization of a transaction as recited in claim 8, wherein said personal information code comprises a code entered with a trusted keypad of said trusted authorization device.","10.A method of providing for a trusted authorization of a transaction as recited in claim 8, wherein said personal information code is responsive to a biometric input from a user entered with a trusted biometric input device.","11.A method of providing for a trusted authorization of a transaction as recited in claim 8, wherein said personal information code is responsive to a signature input from a user entered with a trusted signature input device.","12.A method of providing for a trusted authorization of a transaction as recited in claim 8, wherein said personal information code is responsive to a voice input from a user entered with a trusted microphone.","13.A method of providing for a trusted authorization of a transaction as recited in claim 8, wherein said personal information code is associated with said trusted authorization device.","14.A method of providing for a trusted authorization of a transaction as recited in claim 8, wherein said personal information code comprises a digest of at least one of a biometric input from a user entered with a trusted biometric input device, a signature input from a user entered with a trusted signature input device, and a voice input from a user entered with a trusted microphone.","15.A method of providing for a trusted authorization of a transaction as recited in claim 1, further comprising providing for receiving fourth information from a physical token in possession of a user and incorporating at least a portion of said fourth information in said second information.","16.A method of providing for a trusted authorization of a transaction as recited in claim 15, wherein said physical token is selected from a credit card, a debit card, and a smart card.","17.A method of providing for a trusted authorization of a transaction as recited in claim 16, wherein the operation of receiving fourth information from a physical token comprises reading said fourth information from said physical token using a trusted reader.","18.A method of providing for a trusted authorization of a transaction as recited in claim 17, wherein said fourth information is stored on a magnetic strip of said physical token, and the operation of receiving fourth information from a physical token comprises reading said fourth information from said magnetic strip using a trusted magnetic card reader.","19.A method of providing for a trusted authorization of a transaction as recited in claim 15, further comprising providing for receiving a personal identification code from a user, and incorporating said personal identification code in said second information, wherein said personal information code is associated with said physical token.","20.A method of providing for a trusted authorization of a transaction as recited in claim 15, further comprising providing for receiving a personal identification code from a user, wherein said personal information code is associated with said physical token, the operation of providing for said set of operations by a trusted processor of said trusted authorization device is responsive to whether said personal identification code corresponds to a stored personal identification code stored in said physical token, and said stored personal identification code is associated with an authentic user of said physical token.","21.A method of providing for a trusted authorization of a transaction as recited in claim 1, wherein said random number is a true random number, and said true random number is responsive to a physical process.","22.A method of providing for a trusted authorization of a transaction as recited in claim 1, wherein said second information comprises at least a portion of said first information.","23.A method of providing for a trusted authorization of a transaction as recited in claim 1, further comprising: a. incrementing a transaction counter; and b. incorporating the value of said transaction counter in said second information.","24.A method of providing for a trusted authorization of a transaction as recited in claim 23, further comprising communicating in plaintext to said first computer the value of said transaction counter.","25.A method of providing for a trusted authorization of a transaction as recited in claim 1, further comprising providing for receiving location information from a trusted location device and incorporating said location information in said second information, wherein said location information is related to the location of said trusted authorization device.","26.A method of providing for a trusted authorization of a transaction as recited in claim 25, wherein said trusted location device comprises a trusted GPS receiver.","27.A method of providing for a trusted authorization of a transaction as recited in claim 1, wherein said first encryption process comprises a Data Encryption Standard (DES) cyclic block code (CBC) manipulation detection code (MDC) using said random number as an initial vector.","28.A method of providing for a trusted authorization of a transaction as recited in claim 1, wherein said first encryption process comprises a digest algorithm selected from MD5 and SHA-1.29.A method of providing for a trusted authorization of a transaction as recited in claim 1, wherein said first encryption process comprises a Public Key Infrastructure (PKI) encryption algorithm using a private key that is associated with either said trusted authorization device or a physical token at least temporarily operatively connected to said trusted authorization device, wherein said first encryption process operates on a digest of said second information.","30.A method of providing for a trusted authorization of a transaction as recited in claim 1, wherein said second encryption process comprises a symmetric encryption of said random number using said set of working keys that are stored in said trusted memory of said transaction authorization device.","31.A method of providing for a trusted authorization of a transaction as recited in claim 30, wherein said working keys are generated by a fourth encryption process, and said fourth encryption process is responsive to a second identification code associated with said trusted authorization device, and to a set of rekeying keys stored in said trusted memory of said trusted authorization device.","32.A method of providing for a trusted authorization of a transaction as recited in claim 1, further comprising communicating in plaintext to said first computer at least one encryption algorithm identification code associated with at least one of said first encryption process and said third encryption process.","33.A method of providing for a trusted authorization of a transaction, comprising: a. providing for initiating a transaction on a first computer responsive to at least one input from a user; b. providing for communicating first information to a transaction authorization device, wherein said first information is related to said transaction, and said transaction authorization device is operatively connected to said first computer; c. providing for receiving a data structure from said transaction authorization device, wherein said data structure is responsive to said first information, said data structure comprises a random number, a first identification code, and third information, said third information comprises an encryption by a third encryption process of both second information and a signature responsive to said second information, a first portion of said second information is responsive to said first information, a second portion of said second information comprises said random number, a third portion of said second information comprises said first identification code, said random number is generated by said trusted authorization device, and said first identification code is associated with said trusted authorization device; and d. providing for communicating said data structure to a host server computer, wherein said data structure provides for a trusted authorization of said transaction.","34.A method of providing for a trusted authorization of a transaction, comprising: a. providing for receiving by a first computer a data structure from a second computer, wherein said data structure is responsive to first information, said first information is related to a transaction to be authorized, said data structure comprises a random number, a first identification code, and third information, said third information comprises an encryption by a third encryption process of both second information and a signature by a first encryption process responsive to said second information, a first portion of said second information is responsive to said first information, a second portion of said second information comprises said random number, a third portion of said second information comprises said first identification code; b. providing for retrieving a set of stored working keys, wherein said operation of retrieving is responsive to said first identification code; c. providing for generating a set of session keys by a second encryption process, wherein said second encryption process is responsive to said random number and to said set of stored working keys; d. providing for generating second information and fifth information by decrypting said third information using said third encryption process that is responsive to said set of session keys; e. providing for generating a signature of said second information, wherein said signature is generated by said first encryption process; f. providing for comparing said signature with said fifth information; and g. if said signature matches said fifth information, then providing for acting upon said second information.","35.A method of providing for a trusted authorization of a transaction as recited in claim 25, further comprising providing for transmitting to a third computer said random number, a set of encrypted working keys for said third encryption process and said third information, and receiving from said third computer a result, wherein said set of encrypted working keys is responsive to said first identification code, said set of encrypted working keys are encrypted with a set of keys of said third computer, the operation of providing for acting upon said second information is responsive to said result and said third computer performs the operations of providing for generating said set of session keys, providing for generating said second information and said fifth information, providing for generating said signature of said second information, and providing for comparing said signature with said fifth information.","36.A method of authorizing a transaction responsive to a data structure, comprising: a. receiving said data structure, wherein said data structure is responsive to first information, said first information is related to a transaction to be authorized, said data structure comprises a random number, a first identification code, and third information, said third information comprises an encryption by a third encryption process of both second information and a signature by a first encryption process responsive to said second information, a first portion of said second information is responsive to said first information, a second portion of said second information comprises said random number, a third portion of said second information comprises said first identification code; b. retrieving or receiving a set of stored working keys, wherein said operation of retrieving is responsive to said first identification code; c. generating a set of session keys by a second encryption process, wherein said second encryption process is responsive to said random number and to said set of stored working keys; d. generating second information and fifth information by decrypting said third information using said third encryption process that is responsive to said set of session keys; e. generating a signature of said second information, wherein said signature is generated by said first encryption process; f. comparing said signature with said fifth information; and g. transmitting a result of the operation of comparing said signature with said fifth information.","37.A method of authorizing a transaction responsive to a data structure as recited in claim 36, wherein said set of stored working keys are encrypted, further comprising decrypting said set of stored working keys prior to the operation of generating a set of session keys.","38.A memory for storing data for access by an application program being executed on a computer, comprising a data structure stored in said memory, wherein said data structure is responsive to first information, said first information is related to a transaction to be authorized, and said data structure comprises a. a first data object comprising a random number; b. a second data object comprising a first identification code; and c. a third data object comprising third information, wherein said third information comprises an encryption by a third encryption process of both second information and a signature by a first encryption process responsive to said second information, said third encryption process is responsive to a set of session keys that are responsive to said random number, a first portion of said second information is responsive to said first information, a second portion of said second information comprises said random number, a third portion of said second information comprises said first identification code.","39.A memory for storing data for access by an application program being executed on a computer as recited in claim 38, wherein said data structure further comprises a fourth data object comprising a value of a transaction counter.","40.A memory for storing data for access by an application program being executed on a computer as recited in claim 38, wherein said second information incorporates a value of a transaction counter."],["<SOH> BRIEF DESCRIPTION OF THE DRAWINGS <EOH>In the accompanying drawings: FIG.","1 illustrates a block diagram of a trusted authorization device and an associated transaction processing system; FIG.","2 illustrates a process for providing for a trusted authorization of a transaction from the point-of-view of an associated Trusted Authorization Device (TAD); FIG.","3 illustrates as data structure used by a process for providing for a trusted authorization of a transaction; FIG.","4 a illustrates an encryption process for generating a set of encryption keys; FIG.","4 b illustrates a schematic representation of the process illustrated in FIG.","4 a; FIG.","5 a illustrates 3-DES encryption process for generating a set of encryption keys; FIG.","5 b illustrates a schematic representation of the process illustrated in FIG.","5 a; FIG.","6 a illustrates 3-DES encryption process for encrypting a message; FIG.","6 b illustrates a schematic representation of the process illustrated in FIG.","6 a; FIG.","7 a illustrates 3-DES decryption process for decrypting an encrypted message; FIG.","7 b illustrates a schematic representation of the process illustrated in FIG.","6 a; FIG.","8 illustrates a process for providing for a trusted authorization of a transaction from the point-of-view of an associated client computer; FIG.","9 illustrates a process for providing for a trusted authorization of a transaction from the point-of-view of an associated host computer; FIG.","10 illustrates a process for providing for a trusted authorization of a transaction from the point-of-view of an associated Verification Decryption Server (VDS); FIG.","11 a illustrates a key loading process from the point-of-view of an associated Key Loading Unit (KLU); FIG.","11 b illustrates a schematic representation of the process illustrated in FIG.","11 a; FIG.","12 a illustrates a key loading process from the point-of-view of an associated Trusted Authorization Device; FIG.","12 b illustrates a schematic representation of the process illustrated in FIG.","11 a; FIG.","13 illustrates a re-keying process from the point-of-view of an associated Key Loading Unit (KLU); FIG.","14 illustrates a re-keying process from the point-of-view of an associated Trusted Authorization Device; FIG.","15 illustrates a re-keying process from the point-of-view of an associated Trusted Authorization Device; FIG.","16 illustrates a process for providing for a trusted authorization of a transaction from the point-of-view of an associated Trusted Authorization Device; FIG.","17 illustrates a process for providing for a trusted authorization of a transaction from the point-of-view of an associated Verification Decryption Server (VDS); FIG.","18 illustrates a table of TAD input commands; FIG.","19 illustrates a table of TAD language codes; FIG.","20 illustrates a structure of a TAD input command for authorizing data; FIGS.","21 a illustrates a plaintext portion of a data structure of a TAD response to command for authorizing data; FIG.","21 b illustrates an encrypted portion of a data structure of a TAD response to command for authorizing data; FIG.","22 illustrates a table of data field types associated with a TAD data structure; FIG.","23 illustrates a structure of an error response data packet from a TAD to a client computer if an authorization is either aborted or incorrect; FIG.","24 illustrate a structure of TAD response to command for authorizing data; FIG.","25 illustrate an example of a TAD data structure at various processing stages; FIG.","26 a illustrates a structure of a TAD input command for loading a rekeying keyset; FIG.","26 b illustrates a structure of a TAD response to a command for loading a rekeying keyset; FIG.","27 a illustrates a structure of a TAD input command for installing a new working keyset; FIG.","27 b illustrates a structure of a TAD response to a command for installing a new working keyset; FIG.","28 a illustrates a structure of a TAD input command for installing a new language; FIG.","28 b illustrates a structure of a TAD response to a command for installing a new language; FIG.","29 a illustrates a structure of a TAD input command for identifying a TAD to a client computer; FIG.","29 b illustrates a structure of a TAD response to a command for identifying a TAD to a client computer; FIG.","30 a illustrates a structure of a TAD input command for testing a TAD maintenance key; FIG.","30 b illustrates a structure of a TAD response to a command for testing a TAD maintenance key; FIG.","31 a illustrates a structure of a TAD input command for personalizing a TAD; and FIG.","31 b illustrates a structure of a TAD response to a personalizing a TAD.","detailed-description description=\"Detailed Description\" end=\"lead\"?"],["BRIEF DESCRIPTION OF THE DRAWINGS In the accompanying drawings: FIG.","1 illustrates a block diagram of a trusted authorization device and an associated transaction processing system; FIG.","2 illustrates a process for providing for a trusted authorization of a transaction from the point-of-view of an associated Trusted Authorization Device (TAD); FIG.","3 illustrates as data structure used by a process for providing for a trusted authorization of a transaction; FIG.","4a illustrates an encryption process for generating a set of encryption keys; FIG.","4b illustrates a schematic representation of the process illustrated in FIG.","4a; FIG.","5a illustrates 3-DES encryption process for generating a set of encryption keys; FIG.","5b illustrates a schematic representation of the process illustrated in FIG.","5a; FIG.","6a illustrates 3-DES encryption process for encrypting a message; FIG.","6b illustrates a schematic representation of the process illustrated in FIG.","6a; FIG.","7a illustrates 3-DES decryption process for decrypting an encrypted message; FIG.","7b illustrates a schematic representation of the process illustrated in FIG.","6a; FIG.","8 illustrates a process for providing for a trusted authorization of a transaction from the point-of-view of an associated client computer; FIG.","9 illustrates a process for providing for a trusted authorization of a transaction from the point-of-view of an associated host computer; FIG.","10 illustrates a process for providing for a trusted authorization of a transaction from the point-of-view of an associated Verification Decryption Server (VDS); FIG.","11a illustrates a key loading process from the point-of-view of an associated Key Loading Unit (KLU); FIG.","11b illustrates a schematic representation of the process illustrated in FIG.","11a; FIG.","12a illustrates a key loading process from the point-of-view of an associated Trusted Authorization Device; FIG.","12b illustrates a schematic representation of the process illustrated in FIG.","11a; FIG.","13 illustrates a re-keying process from the point-of-view of an associated Key Loading Unit (KLU); FIG.","14 illustrates a re-keying process from the point-of-view of an associated Trusted Authorization Device; FIG.","15 illustrates a re-keying process from the point-of-view of an associated Trusted Authorization Device; FIG.","16 illustrates a process for providing for a trusted authorization of a transaction from the point-of-view of an associated Trusted Authorization Device; FIG.","17 illustrates a process for providing for a trusted authorization of a transaction from the point-of-view of an associated Verification Decryption Server (VDS); FIG.","18 illustrates a table of TAD input commands; FIG.","19 illustrates a table of TAD language codes; FIG.","20 illustrates a structure of a TAD input command for authorizing data; FIGS.","21a illustrates a plaintext portion of a data structure of a TAD response to command for authorizing data; FIG.","21b illustrates an encrypted portion of a data structure of a TAD response to command for authorizing data; FIG.","22 illustrates a table of data field types associated with a TAD data structure; FIG.","23 illustrates a structure of an error response data packet from a TAD to a client computer if an authorization is either aborted or incorrect; FIG.","24 illustrate a structure of TAD response to command for authorizing data; FIG.","25 illustrate an example of a TAD data structure at various processing stages; FIG.","26a illustrates a structure of a TAD input command for loading a rekeying keyset; FIG.","26b illustrates a structure of a TAD response to a command for loading a rekeying keyset; FIG.","27a illustrates a structure of a TAD input command for installing a new working keyset; FIG.","27b illustrates a structure of a TAD response to a command for installing a new working keyset; FIG.","28a illustrates a structure of a TAD input command for installing a new language; FIG.","28b illustrates a structure of a TAD response to a command for installing a new language; FIG.","29a illustrates a structure of a TAD input command for identifying a TAD to a client computer; FIG.","29b illustrates a structure of a TAD response to a command for identifying a TAD to a client computer; FIG.","30a illustrates a structure of a TAD input command for testing a TAD maintenance key; FIG.","30b illustrates a structure of a TAD response to a command for testing a TAD maintenance key; FIG.","31a illustrates a structure of a TAD input command for personalizing a TAD; and FIG.","31b illustrates a structure of a TAD response to a personalizing a TAD.","DESCRIPTION OF EMBODIMENT(S) Electronic communications, and the data which traverses those communications, are relatively new, as is the technology used to protect electronic data.","Existing communications protection technologies tend to fall into two categories.","The first, government sponsored, is generally very well thought out and provides excellent protection, but is not readily available for commercial applications.","The second, de facto commercial, are mostly not strong enough to protect important information, or are dedicated to specific functions.","For example, standard point-of-sale devices are dedicated to merchandizing applications, and existing ATM systems are dedicated to the dispensing of cash.","There exists a need for a device to provide personal protection of electronic data that is small, easy to use, provides excellent protection to the PC/laptop user, and that can operate in conjunction with corresponding devices at a central data gathering point to provide near real time validation of the information.","As one example, involving financial transactions over the internet by a user, a financial institution may desire an enhanced level of security so as to verify that the user is who they say they are and that they have truly authorized a particular transaction.","As another example, in a business-to-business environment, a paycheck processing company needs to know with virtual certainty the authenticity of instructions from associated business clients for making payroll distributions.","As yet another example, in a gaming environment, a user of internet gaming services may wish to transfer funds from a credit card to a gaming card so as to participate in internet gaming, a transaction for which the credit card issuer generally demands authentication of the user and verification of the transaction so as to avoid a later repudiation of the transaction by the user.","Referring to FIG.","1, a Trusted Authorization Device 10 (or TAD) is operatively connected to an untrustworthy client 12 (for example, a personal computer or workstation PC) operated by a user 14.The TAD 10 provides a trustworthy subsystem that allows the authorization of electronic data transactions or actions while electronically connected to the untrustworthy client 12 platform in a potentially hostile environment.","The TAD 10 comprises a trusted control processor 16, a trusted display 18, a trusted keypad 20 (e.g.","a numeric keypad, or an alphanumeric keyboard), and a trusted device reader 22 (e.g.","a magnetic stripe card, a chip card, a smart cart, etc.).","In one embodiment, the TAD 10 is a computer ancillary hardware device that provides a trusted environment for the display and authorization of transactions/authorizations that can be compactly represented in human understandable form.","The TAD 10 is operatively connected to the client 12 with a telecommunications channel 24, for example, a serial telecommunications (RS-232) interface that is constrained by the trusted control processor 16.A telecommunications channel 24 that is hardwired directly to the client 12 and not out-of-sight therefrom provides for enhanced security.","The trusted control processor 16 and associated memory 26 are, for example, securely packaged in a tamper-proof, hardened housing, that if tampered with causes at least essential elements of the TAD 10 to either self-destruct or become inoperable and virtually undecipherable.","For example, the trusted control processor 16 may be adapted with either a light sensor or a pressure sensor or both, which would cause the memory 26 to be erased response to an associated detection of light or pressure change that would result from tampering with the housing of the trusted control processor 16 and/or associated memory 26.The trusted control processor 16 is a dedicated CPU in the TAD 10 that controls and/or manages the trusted display 18, trusted keypad 20, trusted device reader 22, and the telecommunications channel 24.The trusted control processor 16 provides for the following capabilities: 1.A unique and well protected ID; 2.Embedded unique implementation of cryptographic algorithms (including digest algorithms, asymmetric encryption with public/private keys and symmetric encryption with private/private keys); 3.Random number generation; 4.Functionality to derive unique session keys per transaction from random generation sub process; 5.Cryptographic functionality to enable signing (digest and/or public/private encryption keys) and data encryption (private/private encryption keys) functionality; 6.Ability to protect the TAD encryption keys from observation or alteration.","This involves on-chip hardening and system auto-destruct functionality in event of tamper detection.","Tamper detection sensing is embedded in the TAD 10; 7.Ability to display proposed transactions supplied from host and load supplied information into appropriate data structures for transmission; 8.Ability to capture and load data from integrated TAD devices; 9.Client PC interface, trusted keypad, trusted device reader, e.g.","card reader; 10.Optional camera, biometric input devices, and/or GPS receiver; and 11.Ability to integrate all authorized information into a transaction data structure, digitally sign this data structure, strongly and uniquely encrypt the appropriate portions of the structure, and transmit this data structure to the host for further transmission.","The trusted display 18 is, for example, a separate display constrained by trusted control processor 16 and not subject to intercept, control or modification by a host system.","The trusted display 18 displays the transaction or authorization to be performed, for example in a compact embodiment, on a 4 line by 20 character screen.","The trusted keypad 20 is, for example, a numeric key pad (with alpha functionality) that is constrained by the trusted control processor 16 and whose data is not subject to interception, alteration, or replacement by signals from the client 12.The trusted keypad 20 is used by a user to enter information and accept or refuse a transaction.","The trusted device reader 22 is, for example, a magnetic card/smart card reader that is constrained to communicate with the trusted control processor 16 and whose information is not subject to interception, alteration, or replacement by signals from the client 12.The trusted device reader 22 provides a means for the user 14 to provide proof of possession or the associated magnetic card/smart card, and thereby enable the TAD 10 to authenticate the transaction request.","The trusted device reader 22 may, for example, be a hybrid card reader, enabling it to support the usage of chip cards in a trustworthy environment.","Such chip cards provide an appropriate environment for accessing user-specific public key-enabled functionality.","The client 12 is operatively connected, e.g.","via the Internet, to a host server 28 having a communication interface 30.For example, the host server 28 could be operated by a service provider that requires an enhanced level of trust in the authorization of transactions or requests by the user 14 running particular application software of the service provider, and accordingly, who would provide a TAD 10 to the user 14 for authenticating transactions with the necessary enhanced level of trust.","The host server 28 interfaces with a verification decryption server 32 (VDS), for example, via a VDS executive manager 34, and may also, or alternatively, interface with a customer application system 36 running associated application software, and also interfaced with the VDS 32.Each TAD 10 is provided with a unique alphanumeric ID (TADID_A) and a unique and well-protected binary ID (TADID_B), each of which are stored in memory 26.The alphanumeric ID (TADID_A) is also visible on the outside of an associated housing of the TAD 10 for purposes of identifying the particular device, for example for purposes of maintenance or physical distribution control.","The associated trusted control processors 16 are also provided with embedded unique implementations of cryptographic algorithms, including at least one algorithm for generating a signature—which may include a digest process—(e.g.","asymmetric encryption with public/private keys) and at least one algorithm for encrypting data (e.g.","symmetric encryption with private/private keys), the later of which relies upon associated keys that are stored in the TAD 10 by a key loading unit 38 (KLU).","Referring to FIG.","2, in operation, in step (202) the untrustworthy client 12 downloads a proposed transaction or action to the TAD 10 for authorization, for example at the request of the user 14.Then, in step (204), the user 14, viewing the trusted display 18 and using the trusted keypad 20, pages through the proposed transaction, thereby providing a basis of trust for the action to be authorized.","When the user 14 wishes to authorize the transaction, the user 14 can accept the transaction by pressing the appropriate key, or sequence of keys, on the trusted keypad 20.The user 14 may then further couple a unique physical token 40—e.g.","a magnetic stripe card 40.1 or a smart card 40.2—to the trusted device reader 22, and then enter a personal PIN number associated therewith on the trusted keypad 20, thereby authenticating the identity of the user 14.Alternately, or in addition, the TAD 10 may require the user 14 to enter a PIN number associated with the TAD 10.Then, in step (206), if the user 14 authorizes the transaction, then the trusted control processor 16 stores the displayed first information, the captured card information, and all other necessary information (e.g.","PIN, location from an associated trusted location device e.g.","GPS receiver, etc) as second information in an associated data fields of an associated data structure 42, e.g.","illustrated in FIG.","3, wherein the second information is responsive to, or a function of the first information, and may comprise a copy of the first information.","Otherwise, from step (206), the process repeats with step (202), wherein the TAD 10 awaits further input from the client 12.Then, in step (208), a random number generator 44—e.g.","within the trusted control processor 16—generates a random number R, which may be either a pseudo-random number, or a true random number, for example, generated responsive to a noisy physical process.","The TAD 10 may also access and increment a transaction counter, although this step is not essential.","Then, in step (210), the trusted control processor 16 generates second information that is responsive to the first information displayed to the user, and which further incorporates the random number R and a first identification code of the TAD 10, e.g.","the alphanumeric ID (TADID_A).","Then, in step (212) the trusted control processor 16 generates a digital signature of the second information—thereby providing a basis for authorization and non-repudiation of the transaction—using a first encryption process, for example, an irreversible digest algorithm (e.g.","an asymmetric encryption algorithm).","Then, in steps (214) and (216), the trusted control processor 16 respectively retrieves a set of stored working keys Kw1, Kw2, Kw3 from the memory 26, and generates a set of transaction-specific session keys Ks1, Ks2, Ks3 using a second encryption process—generally illustrated in FIGS.","4a, 4b, and illustrated for a 3-DES encryption process in FIGS.","5a and 5b—using the random number R as a seed.","The working keys Kw1, Kw2, Kw3 are stored in the memory 26 by a key loading process described hereinbelow.","Then, in step (218), trusted control processor 16 generates third information by encrypting the combination of the second information from step (210), and the associated signature from step (212), using a third encryption process, for example a 3-DES encryption process as illustrated in FIGS.","6a and 6b.","Then, in step (220), the data structure 42 comprising the plaintext random number R, the plaintext alphanumeric ID (TADID_A), and the third information is communicated to the client 12, and communicated thereby to the host server 28.The host server 28 sends the data structure 42 to the VDS 32 for decryption and signature verification thereby, and if authenticated by the VDS 32, the transaction is processed by either the host server 28 or the associated customer application system 36.If a transaction counter is used, the value of the transaction counter would be incorporated in the second information (which is signed), and may also be incorporated as plaintext in the data structure 42.For example, if the user is interfaced with the host server 28 is via an Internet browser, a user may select, via the browser, a transaction to be conducted—for example, the transfer of funds from an account accessed via a standard financial card or a transaction within a custom domain.","The browser then uses associated TAD 10 interface software to transfer the proposed transaction and instructions to the TAD 10 for authorization.","Referring to FIG.","8, from the point of view of the client 12, the above described process commences in step (802), wherein the user 14 initiates a transaction on a client 12 in communication with the host server 28.For example, the user 14 initiates a transaction on the Internet involving a purchase that the user 14 wishes to finance by a credit card, i.e.","a magnetic stripe card 40.1.In step (804), responsive to the host server 28 requesting a trusted authorization of the transaction, a first information to be authorized is communicated to the TAD 10, and in step (806), if the user 14 has authorized the transaction using the TAD 10, the client 12 receives the associated data structure 42 from the TAD 10, and, in step (808), communicates this to the host server 28.Referring to FIG.","9, from the point of view of the host server 28, in step (902), the host server 28 receives the data structure 42 from the client 12, in step (904), extracts the alphanumeric ID (TADID_A) from the plaintext portion of the data structure 42 and uses a lookup process to find an associated set of VDS encrypted TAD working keys KVDS—w1, KVDS—w2, KVDS—w3 that are encrypted using a VDS key that is not known by the host server 28.Then, in step (906), the VDS encrypted TAD working keys KVDS13 w1, KVDS—w2, KVDS—W3 and the data structure 42 are communicated by the host server 28 to the VDS 32, responsive to which, in step (908), the host server 28 receives a verification status from the VDS 32, and in step (910), the host server 28 communicates this verification status to the client 12.Referring to FIG.","10, from the point of view of the verification decryption server 32 (VDS), in step (1002), the verification decryption server 32 receives the VDS encrypted TAD working keys KVDS—w1, KVDS—w2, KVDS—w3 and the data structure 42 from the host server 28.Then, in step (1004), the VDS 32 decrypts the VDS encrypted TAD working keys KVDS—w1, KVDS—w2, KVDS—w3 using associated VDS keys KVDS1, KVDS2, KVDS3 stored on the VDS 32 and loaded thereon by the key loading unit 38.Then, in step (1006), the VDS 32 extracts the plaintext random number R from the data structure 42, and in step (1008), uses the random number R as a seed, together with the decrypted TAD working keys Kw1, Kw2, Kw3 from step (1004) to generate—by a key generating process 500 as illustrated in FIGS.","5a and 5b—a set of session keys Ks1, Ks2, Ks3 that are used in step (1010) to decrypt the encrypted portion (i.e.","the second information) of the data structure 42, for example, in accordance with a 3-DES decryption process as illustrated in FIGS.","7a and 7b.","Then, in step (1012), the VDS 32 generates a signature of the second information using the same first encryption process as had been used in the TAD 10, and in step (1014), if the extracted, decrypted signature from step (1010) is the same as the generated signature from step (1012), then in step (1016), the verification process is successful, and in steps (1018) and (1020), the information responsive to the first information is extracted from the second information, and then communicated to the customer application system 36 to complete the transaction, after which the customer application system 36 notifies the host server 28 that the transaction has been completed successfully.","Then, in step (1022), the verification status is communicated by the VDS 32 to the host server 28.If, from step (1014), the generated signature is not equal to the extracted, decrypted signature, then, in step (1024), the verification process is unsuccessful, and this verification status is communicated to the host server 28 in step (1022).","The TAD 10 incorporates three sets of stored keys that are used in various encryption processes, 1) a set of read-only firmware keys KF1, KF2, KF3, 2) a set of rekeying keys KRK1, KRK2, KRK3 that are initially loaded on the TAD 10 by the key loading unit 38, and 3) a set of working keys Kw1, Kw2, Kw3 that are loaded on the TAD 10 during a rekeying operation by the key loading unit 38, either directly connected to the TAD 10, or remotely via a mailed floppy disk.","The TAD 10 uses the working keys Kw1, Kw2, Kw3 to generate the transaction-specific session keys Ks1, Ks2, Ks3 in accordance with a key generating process 500, as described hereinabove.","The key loading unit 38 loads the rekeying keys KRK1, KRK2, KRK3 and the working keys Kw1, Kw2, Kw3 on the TAD 10 by transferring an encrypted key seed to the TAD 10, after which the TAD 10 generates the respective keys using a key generating process 500, in which the TAD 10 and the key loading unit 38 each utilize prearranged secret keys to encrypt the key seed.","For example, both binary ID (TADID_B) and the firmware keys KF1, KF2, KF3 are known to both the TAD 10 and the key loading unit 38, wherein the key loading unit 38 is able to determine the binary ID (TADID_B) from a lookup process, given the alphanumeric ID (TADID_A) of the TAD 10.Accordingly, both the key loading unit 38 and the TAD 10 can independently use the key generating process 500—with the binary ID (TADID_B) as the seed (with S1=S2=S3=TADID_B) and the firmware keys KF1, KF7, KF3 as the generating keys—to generate a set of maintenance keys maintenance keys KM1, KM2, KM3.The key loading unit 38 then uses the maintenance keys KM1, KM2, KM3 to encrypt a rekey random number RkR, which is then transferred in encrypted form to the TAD 10, which then decrypts the rekey random number RkR and uses this as a seed, together with the maintenance keys KM1, KM2, KM3 as generating keys, to generate the rekeying keys KRK1, KRK2, KRK3.Similarly, the key loading unit 38 can use the same rekey random number RkR as a seed, and the same maintenance keys KM1, KM2, KM3 as generating keys, to independently generate identical rekeying keys KRK1, KRK2, KRK3.Then, the key loading unit 38 can use the rekeying keys KRK1, KRK2, KRK3 to encrypt a working key random number that is transferred to the TAD 10 to be used thereby as a seed in accordance with the key generating process 500 to generate the TAD working keys Kw1, Kw2, Kw3, wherein the rekeying keys KRK1, KRK2, KRK3 are used as associated key generating keys.","The key loading unit 38 is used to load the VDS encrypted TAD working keys KVDS—w1, KVDS—w2, KVDS—w3 on the VDS 32, using VDS keys for the encryption, after which the TAD rekeying keys KRK1, KRK2, KRK3 and TAD working keys Kw1, Kw2, Kw3 are destroyed on the key loading unit 38 so that the key loading unit 38 cannot become a single point source of security failure.","The process by which the key loading unit 38 transfers and encrypted key seed to the TAD 10 is illustrated in FIG.","11a, and is represented schematically in FIG.","11b.","The process by which the TAD 10 receives the encrypted key seed from the key loading unit 38 and decrypts the key seed is illustrated in FIG.","12a, and is represented schematically in FIG.","12b.","The process by which respective key seeds for the rekeying keys KRK1, KRK2, KRK3 and the working keys Kw1, Kw2, Kw3 are respectively generated and encrypted by key loading unit 38, and transferred to the TAD 10, is illustrated in FIG.","13.The process by which respective key seeds for the rekeying keys KRK1, KRK2, KRK3 and the working keys Kw1, Kw2, Kw3 are received by the TAD 10 and used to generate the respective rekeying keys KRK1, KRK2, KRK3 and working keys Kw1, Kw2, Kw3, is illustrated in FIG.","14.Referring also to FIG.","15, the key loading unit 38 provides for manual key distribution and management services using a limited set of commands.","The TAD uses the maintenance key to decrypt the re-keying keys.","This operation should be done locally via a direct connection to the key loading unit.","The TAD relies upon a set of 3 DES re-keying keys to load the unit keys that the unit relies upon.","These re-keying keys are installed in the TAD 10 by the key loading unit 38 via the TAD maintenance keys, which are internally generated in the TAD by the interaction of the TAD firmware keyset (which is common to all the TAD's in a production lot) with a TAD-specific 64 bit Binary ID, TADID_B.","The three maintenance keys are generated by permuting the order of the firmware keyset in a triple DES EDE encryption of the BID, i.e.","Km1=EKf1(DKf2(EKf3(TADID—B))), Km2=EKf3(DKf2(EKf1(TADID—B))), Km3=EKf2(DKf1(EKf3(TADID—B))), (wherein E and D respectively represent the encryption and decryption sub-processes of a symmetric encryption process, e.g.","triple DES (3 DES).","Kf1 is the first firmware key.","Km1 is the first maintenance key.","etc.","Using the above processes the TAD can generate the TAD-specific maintenance keyset.","Similarly, the keyloader, which knows the firmware keys and the BID can also generate and use the TAD-specific maintenance keyset.","Thus, the keyloader can load the re-keying keys into the TAD.","Using the maintenance keys Km1, Km2, Km3, the re-keying keys KRK1, KRK2, KRK3 are generated therein by execution of a load re-key command (a rekey command with no re-keying keys is assumed to be a load re-key command), having a command structure is: Clear text portion Re-key Command 4 bytes (0XFFFFFFFF) Re-keying block length 4 bytes (multiple of 8 bytes) Random number Rk 24 bytes Encrypted portion Random number length 4 Re-keying random number as specified (initially 24) digest prior to encryption as appropriate The size of the structures may vary with system versions.","The encryption keys in process are the maintenance keys Km1, Km2, Km3.Upon receiving a re-key command with no re-keying keys, the TAD 10 performs the following functions: 1.Check to make sure that the block is the right length (if wrong, return a failure code and clear the block); 2.Calculate the session keys, using the random number Rk provided in the clear text portion, as follows: a.","Take the first 8 bytes of the session_random number, R1_and calculate session key 1, Ks1=EKm1(DKm2(EKm3(R1))) (wherein E and D respectively represent the encryption and decryption sub-processes of a symmetric encryption process, e.g.","triple DES (3 DES); b.","Take the second 8 bytes of the session_random number, R2_and calculate session key 2, Ks2=EKm3(DKm2(EKm1(R2))); and c. Take the third 8 bytes of the session random number, R3 and calculate session key 3, Ks3=EKm2(DKm1(EKm3(R3))); 3.Decrypt the encrypted portion DKs3(Es2(DKs1(encrypted portion))) using CBC mode; 4.Calculate the MD5 hash of the entire block cleartext+decrypted portion (digest block set to NULL); 5.Compare calculated digest with received digest (if wrong, return a failure code and clear the block); 6.Take the first 8 bytes of the re-keying random number, RKR1 and calculate re-keying key 1, KrK1=EKm1(DKm2(EKm3(RkR1))); 7.Take the second 8 bytes of the re-keying random number, RkR2 and calculate re-keying key 2, KrK2=Ekm3(DKm2(EKm1(RkR2))); and 8.Take the third 8 bytes of the re-keying random number, RkR3 and calculate re-keying key 3, KrK3=EKm2(DKm1(EKm3(RkR3))).","The re-keying command can be issued after the re-keying keys Kr1, Kr2, Kr3 have been installed in the TAD 10 by the key loading unit 38, wherein the key loading unit 38 calculates the re-keying keys Kr1, Kr2, Kr3 from the binary ID (TADID_B).","After the re-keying keys Kr1, Kr2, Kr3 are installed in the TAD 10 by the key loading unit 38, the working keys Kw1, Kw2, Kw3 are generated therein by execution of a re-key command, having a command structure is: Plaintext portion Re-key Command 4 bytes (0XFFFFFFFF) Re-keying block length 4 bytes (multiple of 8 bytes) Random number Rk 24 bytes Encrypted portion Re-keying counter 4 Random number length 4 Re-keying random number as specified (initially 24) digest prior to encryption as appropriate The associated re-keying data structure, for example, does not have the flexibility of the general data structures.","For example, the key loading unit 38 knows what algorithms are expected by the TAD 10 and uses the appropriate ones, hence there is no need for flexibility here.","The size of the structures may vary with system versions.","The encryption keys in process are not the working keys Kw1, Kw2, Kw3, but instead are the re-keying keys Kr1, Kr2, Kr3.Upon receiving a re-key command, the TAD 10 performs the following functions: 1.Check to make sure that the block is the right length (if wrong, return a failure code and clear the block); 2.Calculate the session keys, using the session random number Rk provided in the plaintext portion, as follows: a.","Take the first 8 bytes of the session random number, R1, and calculate session key 1, Ks1: Ks1=EKr1(DKr2(Er3(R1))) (wherein E and D respectively represent the encryption and decryption sub-processes of a symmetric encryption process, e.g.","triple DES (3 DES); b.","Take the second 8 bytes of the session random number, R2, and calculate session key 2, Ks2: Ks2=EKr3(DKr2(EKr1(R2))); and c. Take the third 8 bytes of the session random number, R3 (alternately, R3 could be generated from R1 and R2 by R3=R1 XOR R2), and calculate session key 3, Ks3: Ks3=EKr2(DKr1(EKr3(R3))).","3.Decrypt the encrypted portion DKs3(EKs2(DKs1(encrypted portion))) using CBC mode; 4.Calculate the MD5 hash of the entire block plaintext+decrypted portion (digest block set to NULL); 5.Compare calculated digest with received digest (if wrong, return a failure code and clear the block); 6.Check re-keying counter (which is initially set to 0).","If less than current re-keying counter, return a failure code and clear the block; 7.Set re-keying counter and compute net unit working keys; 8.Take the first 8 bytes of the re-keying random number, RkR1 and calculate working key 1, Kw1: Kw1=EKr1(DKr2(EKr3(RkR1))); 9.Take the second 8 bytes of the re-keying random number, RkR2 and calculate working key 2, Kw2: Kw2=EKr3(DKr2(EKr1(RkR2))); and 10.Take the third 8 bytes of the re-keying random number, RkR3 (alternately, R3 could be generated from R1 and R2 by R3=R1 XOR R2), and calculate working key 3, Kw3: Kw3=EKr2(DKr1(EKr3(RkR3))).","After the working keys Kw1, Kw2, Kw3 are initially loaded by the key loading unit 38, the TAD 10 is placed in service proximate to the client 12 for providing trusted signing and authorization of transactions.","The one or more maintenance test keys are used for diagnosis and maintenance of the TAD 10, but generally not for purposes of data encryption.","For example, in a diagnostic mode, the working keys are replaced with the maintenance keys, and are used to encode a dummy transaction, which can then be remotely decoded by maintenance personal to check that the TAD 10 is operating properly.","The data encryption process utilizes a random number R generated by a random number generator 44 within the trusted control processor 16 as a seed for an irreversible digest process (e.g.","an asymmetric encryption process), e.g.","MD5, that signs a portion of token to be encrypted, and in combination with working keys Kw1, Kw2, Kw3 created by a separate key loading process and stored in memory 26, to generate a set of session keys Ks1, Ks2, Ks3 that are used in a symmetric encryption process, e.g.","triple DES (3 DES) using cyclic block chaining (CBC) mode, to encrypt the signed message.","The trusted control processor 16 also has a set of re-keying keys Kr1, Kr2, Kr3 that are generated by the key loading unit 38 in direct connection with the TAD 10 and stored in memory 26.The re-keying keys Kr1, Kr2, Kr3 thereafter enable the working keys Kw1, Kw2, Kw3 to be update remotely, for example, with new working keys Kw1, Kw2, Kw3 provided to the user 14 over a controlled path, e.g.","via a floppy disc provided by mail or courier.","Referring to FIGS.","18, 19, 20, 26a, 27a, 28a, 29a, 30a, and 31a, the TAD 10 is controlled responsive to various types of TAD input commands that are communicated to the TAD 10 either by the client 12 or the key loading unit 38 over the telecommunications channel 24.FIGS.","20 illustrates the syntax of a command by a client 12 to authorize the signing and encryption of data in accordance with the normal operation of the TAD 10.FIG.","26a illustrates the syntax of a command by a client 12 or the key loading unit 38 to re-key the TAD 10.FIG.","28a illustrates the syntax of a command by a client 12 or the key loading unit 38 to insert a new language in the TAD 10, by which messages are displayed on the trusted display 18.For example, the TAD 10 is configured for selectable predefined languages of English, French, German or Spanish, and is provided with the capability of loading other languages/character sets, e.g.","of Asian languages.","The TAD 10 has a straight forward interface with the client 12, for example, supporting the following three commands from the client: Identify—which returns a text string, the TAD ID#, and the TAD version number.","Process_Transaction—which accepts a proposed transaction, if successful, returning a transaction packet Secure_Transaction—The TAD will display the commands to initiate the card swipe, transaction digest and encryption.","The TAD command interface is as follows: Command 4 bytes Data length 4 bytes Data as specified If no data or result is present, the length is set to 0.Referring to FIG.","16, the TAD 10 provides trusted authorization of transactions in accordance with the following process: 1.Client downloads transaction (a compact, <80 character string) to the TAD 10 in accordance with the command structure illustrated in FIG.","5a.","2.User observes displayed transaction and verifies that it is correct.","a.","If so, the user presses the GO/ACCEPT key.","b.","If not, the user presses the STOP/REJECT key.","3.If rejected, the TAD 10 signals the client and returns a “reject” code.","4.If accepted, the TAD 10 displays the downloaded instructions to the user.","5.The user following the instruction, a. swipes/enters the specified card, b. enters their secret PIN and, c. presses the GO/ACCEPT key, d. Pressing the STOP/REJECT key clears the transaction.","6.If rejected, the TAD 10 signals the client and returns a “reject” code.","7.If the transaction was accepted, the TAD 10 a. digitally signs the captured card data and the PIN, b. packages this information with the transaction, c. appropriately encrypts the sensitive data, and d. prepares the transaction packet 8.The TAD 10 notifies the client of success and transfers the prepared transaction packet, or TAD Output Token, to the client, structured as indicated in FIGS.","21a, 21b, and 24, and by example in FIG.","25.The data is read in wire order.","The TAD 10 is not limited to a particular type of encryption.","The following table indicates an example of various algorithms that can be used for signing and encryption: Packet Digest Identifier 0D 3 DES CBC digest using packet working key set 1D MD5 2D MD160 3D SHA1 Packet Encryption Algorithm ID 0E 3 DES CBC using packet working key set 1E 1 DES using key 1 of packet working key set The client 12 then transmits this to the appropriate host server 28.The transaction generated by the TAD 10 will pass through the communications network and then received by a host server 28 at the customer's site.","The host server 28 will contain the alphanumeric ID (TADID_A) identification number, and corresponding unique working keys Kw1, Kw2, Kw3 in an encrypted form.","When the host server 28 receives a transaction from the client 12 operated by the user 14, the host uses the alphanumeric ID (TADID_A) to retrieve the associated encrypted working keys Kw1, Kw2, Kw3.The host server 28 passes the encrypted working keys Kw1, Kw2, Kw3 and the encrypted transaction to the verification decryption server 32, thereby operating in a stateless mode.","The VDS 32 does not have to store or manage working keys Kw1, Kw2, Kw3 (although for performance reasons one would expect it to cache previously supplied keys).","Referring to FIG.","17, the VDS 32 initially operated as a single-threaded process operating entirely in memory, writing an audit log to hard disc.","Once the VDS 32 accepts a transaction it completes the specified decryption and verification and returns the result before accepting another transaction.","The host server 28 shall poll the VDS 32 before submitting a transaction for verification.","The processes handled by the VDS 32, for example, includes the following: Read the supplied data block Read TAD ID Look up decryption key value Parse encrypted key data structure A VDS 32 has the decryption keys needed to decrypt the encrypted working keys Kw1, Kw2, Kw3.The host server 28 passes the encrypted working keys Kw1, Kw2, Kw3 to the VDS 32, with the following data format, for example, having a structure that is more general than that required to handle to initial implementation of 3 DES keys so as to be able to handle different secret or public key algorithms, as appropriate: Plaintext portion Element Identifier Element Length TAD Unit ID 8 bytes Key structure length 4 bytes Packet Digest Identifier 2 bytes Packet Encryption Algorithm ID 2 bytes Random number 16 bytes Encrypted portion Element Identifier Element Length TAD keys packet length − (32 + digest) digest prior to encryption as appropriate The duplication of databases may be prevented by storing the wrapped TAD keys with the TAD and user data in the host server 28.The use of two sets of unique DES key triplets: the unit re-keying key set, and the unit working key set, helps to reduce the processing and storage requirements that would otherwise be required with public key technology.","The unit's re-keying key set is the basic key set for the unit.","It can only be used with a single command, an encrypted command generated by the keying/re-keying server that causes the TAD 10 to generate the working keys Kw1, Kw2, Kw3 that are used in all security operations.","The key loading unit 38 and host server 28 are trusted with the either the binary ID (TADID_B) or the working keys Kw1, Kw2, Kw3, an accordingly are, for example, implemented so as to provide substantial assurance of the integrity of the storage and processing of these keys, for example, based upon Getronics (formerly Wang) STOP platforms (NSA evaluated B3).","The associated communications protection devices are examined by trusted third parties so as to provide independent assurance concerning the device properties and characteristics.","The TAD 10 has been adapted to incorporate other devices into its trust perimeter such as a GPS receiver 46 and may be adapted to incorporate other devices, such as a signature input device 48, one or more biometric input devices 50 (e.g.","voice, fingerprint, retinal scan), a camera, a breath analyzer 52, and etc.","Referring to FIG.","1, the security and authenticity of the TAD 10 may be with a GPS receiver 46, so as to enable a service provider to ascertain that the user 14 is at the correct prearranged physical location for the TAD 10, and that the TAD 10 has not been relocated.","This is useful in certain business authorizations, where a TAD 10 is issued to a specific customer at a specific location.","The use of the GPS receiver 48 allows the management system to provide strong assurance of the user's location, as well as provide an accurate time value (e.g.","accurate to a millisecond or better).","The coordinates (latitude, longitude, and/or altitude) from the GPS receiver 48 are stored in an associated field, e.g.","Field #5 of the TAD Output data structure illustrated in FIG.","24, and are therefore signed and encrypted along with the rest of the encrypted portion of the TAD Output data structure.","Accordingly, the service provider include a test of the decrypted position coordinates of the TAD 10 from the GPS receiver 46 in deciding whether or not to carry out the transaction requested by the user 14.Whereas the TAD 10 is illustrated as a separate device, it should be understood that the TAD 10 could be embedded in the client 12.Furthermore, whereas the key loading unit 38 is illustrated as a single workstation within the trusted environment of the host server 28 and verification decryption server 32, it should be understood that the key loading unit 38 could be constructed as a portable unit that can be moved to the site of the TAD 10 by a trusted representative.","While specific embodiments have been described in detail, those with ordinary skill in the art will appreciate that various modifications and alternatives to those details could be developed in light of the overall teachings of the disclosure.","Accordingly, the particular arrangements disclosed are meant to be illustrative only and not limiting as to the scope of the invention, which is to be given the full breadth of the appended claims, and any and all equivalents thereof."]],"string":"[\n [\n \"Trusted authorization device\",\n \"A trusted display (18) of a trusted authorization device (TAD) (10) displays on a trusted display (18) first information about a transaction to be authorized by a user (14) using a trusted keypad (20).\",\n \"The TAD (10) generates (208) a random number (R); generates (1210) second information from the first information, the random number (R) and a first identification code (TADID-A) of the TAD (10); generates (212) a signature of the second information using a first encryption process; egnerates (216) a set of session keys (Ks1, Ks2, Ks3) by a second encryption process responsive to the random number (R) and a set of stored working keys (Kw1, Kw2, Kw3); and generates (218) third information by encrypting the second information and the signature using a third encryption process responsive to the set of session keys (Ks1, Ks2, Ks3).\",\n \"A dat structure (42) is formed comprising the random numer (R), the first identification code (TADID-A), and the third information; and communicated (220) from the TAD (10) to the client (12) to a host server (28) for verification by a verification decryption server (32).\"\n ],\n [\n \"1.A method of providing for a trusted authorization of a transaction, comprising: a. providing for communicating with a first computer; b. providing for displaying first information to be authorized on a trusted display of a trusted authorization device, wherein said first information to be authorized is provided by said first computer; c. providing for receiving an authorization command from a trusted keypad of said trusted authorization device, wherein said authorization command is related to said first information; and d. if said authorization command provides for authorizing said first information, then providing for a set of operations by a trusted processor of said trusted authorization device, said set of operations comprising: i. generating a random number; ii.\",\n \"generating second information that is responsive to said first information to be authorized, wherein said second information further incorporates both said random number and a first identification code associated with said trusted authorization device, wherein said first identification code is stored on a trusted memory of said trusted authorization device; iii.\",\n \"generating a signature of said second information, wherein said signature is generated by a first encryption process; iv.\",\n \"generating a set of session keys by a second encryption process, wherein said second encryption process is responsive to said random number and to a set of stored working keys, and said set of stored working keys are stored on said trusted memory of said trusted authorization device; v. generating third information by encrypting said second information and said signature using a third encryption process that is responsive to said set of session keys; and vi.\",\n \"communicating to said first computer said random number, said first identification code, and said third information, wherein said random number and said first identification code are communicated in plaintext.\",\n \"2.A method of providing for a trusted authorization of a transaction as recited in claim 1, further comprising providing for receiving a personal identification code from a user, wherein said personal identification code is incorporated in said second information.\",\n \"3.A method of providing for a trusted authorization of a transaction as recited in claim 2, wherein said personal information code comprises a code entered with a trusted keypad of said trusted authorization device.\",\n \"4.A method of providing for a trusted authorization of a transaction as recited in claim 2, wherein said personal information code is responsive to a biometric input from a user entered with a trusted biometric input device.\",\n \"5.A method of providing for a trusted authorization of a transaction as recited in claim 2, wherein said personal information code is responsive to a signature input from a user entered with a trusted signature input device.\",\n \"6.A method of providing for a trusted authorization of a transaction as recited in claim 2, wherein said personal information code is responsive to a voice input from a user entered with a trusted microphone.\",\n \"7.A method of providing for a trusted authorization of a transaction as recited in claim 2, wherein said personal information code comprises a digest of at least one of a biometric input from a user entered with a trusted biometric input device, a signature input from a user entered with a trusted signature input device, and a voice input from a user entered with a trusted microphone.\",\n \"8.A method of providing for a trusted authorization of a transaction as recited in claim 1, further comprising providing for receiving a personal identification code from a user, wherein the operation of providing for said set of operations by a trusted processor of said trusted authorization device is responsive to whether said personal identification code corresponds to a stored personal identification code, said stored personal identification code is stored in said trusted memory of said trusted authorization device, and said stored personal identification code is associated with an authentic user of said trusted authorization device.\",\n \"9.A method of providing for a trusted authorization of a transaction as recited in claim 8, wherein said personal information code comprises a code entered with a trusted keypad of said trusted authorization device.\",\n \"10.A method of providing for a trusted authorization of a transaction as recited in claim 8, wherein said personal information code is responsive to a biometric input from a user entered with a trusted biometric input device.\",\n \"11.A method of providing for a trusted authorization of a transaction as recited in claim 8, wherein said personal information code is responsive to a signature input from a user entered with a trusted signature input device.\",\n \"12.A method of providing for a trusted authorization of a transaction as recited in claim 8, wherein said personal information code is responsive to a voice input from a user entered with a trusted microphone.\",\n \"13.A method of providing for a trusted authorization of a transaction as recited in claim 8, wherein said personal information code is associated with said trusted authorization device.\",\n \"14.A method of providing for a trusted authorization of a transaction as recited in claim 8, wherein said personal information code comprises a digest of at least one of a biometric input from a user entered with a trusted biometric input device, a signature input from a user entered with a trusted signature input device, and a voice input from a user entered with a trusted microphone.\",\n \"15.A method of providing for a trusted authorization of a transaction as recited in claim 1, further comprising providing for receiving fourth information from a physical token in possession of a user and incorporating at least a portion of said fourth information in said second information.\",\n \"16.A method of providing for a trusted authorization of a transaction as recited in claim 15, wherein said physical token is selected from a credit card, a debit card, and a smart card.\",\n \"17.A method of providing for a trusted authorization of a transaction as recited in claim 16, wherein the operation of receiving fourth information from a physical token comprises reading said fourth information from said physical token using a trusted reader.\",\n \"18.A method of providing for a trusted authorization of a transaction as recited in claim 17, wherein said fourth information is stored on a magnetic strip of said physical token, and the operation of receiving fourth information from a physical token comprises reading said fourth information from said magnetic strip using a trusted magnetic card reader.\",\n \"19.A method of providing for a trusted authorization of a transaction as recited in claim 15, further comprising providing for receiving a personal identification code from a user, and incorporating said personal identification code in said second information, wherein said personal information code is associated with said physical token.\",\n \"20.A method of providing for a trusted authorization of a transaction as recited in claim 15, further comprising providing for receiving a personal identification code from a user, wherein said personal information code is associated with said physical token, the operation of providing for said set of operations by a trusted processor of said trusted authorization device is responsive to whether said personal identification code corresponds to a stored personal identification code stored in said physical token, and said stored personal identification code is associated with an authentic user of said physical token.\",\n \"21.A method of providing for a trusted authorization of a transaction as recited in claim 1, wherein said random number is a true random number, and said true random number is responsive to a physical process.\",\n \"22.A method of providing for a trusted authorization of a transaction as recited in claim 1, wherein said second information comprises at least a portion of said first information.\",\n \"23.A method of providing for a trusted authorization of a transaction as recited in claim 1, further comprising: a. incrementing a transaction counter; and b. incorporating the value of said transaction counter in said second information.\",\n \"24.A method of providing for a trusted authorization of a transaction as recited in claim 23, further comprising communicating in plaintext to said first computer the value of said transaction counter.\",\n \"25.A method of providing for a trusted authorization of a transaction as recited in claim 1, further comprising providing for receiving location information from a trusted location device and incorporating said location information in said second information, wherein said location information is related to the location of said trusted authorization device.\",\n \"26.A method of providing for a trusted authorization of a transaction as recited in claim 25, wherein said trusted location device comprises a trusted GPS receiver.\",\n \"27.A method of providing for a trusted authorization of a transaction as recited in claim 1, wherein said first encryption process comprises a Data Encryption Standard (DES) cyclic block code (CBC) manipulation detection code (MDC) using said random number as an initial vector.\",\n \"28.A method of providing for a trusted authorization of a transaction as recited in claim 1, wherein said first encryption process comprises a digest algorithm selected from MD5 and SHA-1.29.A method of providing for a trusted authorization of a transaction as recited in claim 1, wherein said first encryption process comprises a Public Key Infrastructure (PKI) encryption algorithm using a private key that is associated with either said trusted authorization device or a physical token at least temporarily operatively connected to said trusted authorization device, wherein said first encryption process operates on a digest of said second information.\",\n \"30.A method of providing for a trusted authorization of a transaction as recited in claim 1, wherein said second encryption process comprises a symmetric encryption of said random number using said set of working keys that are stored in said trusted memory of said transaction authorization device.\",\n \"31.A method of providing for a trusted authorization of a transaction as recited in claim 30, wherein said working keys are generated by a fourth encryption process, and said fourth encryption process is responsive to a second identification code associated with said trusted authorization device, and to a set of rekeying keys stored in said trusted memory of said trusted authorization device.\",\n \"32.A method of providing for a trusted authorization of a transaction as recited in claim 1, further comprising communicating in plaintext to said first computer at least one encryption algorithm identification code associated with at least one of said first encryption process and said third encryption process.\",\n \"33.A method of providing for a trusted authorization of a transaction, comprising: a. providing for initiating a transaction on a first computer responsive to at least one input from a user; b. providing for communicating first information to a transaction authorization device, wherein said first information is related to said transaction, and said transaction authorization device is operatively connected to said first computer; c. providing for receiving a data structure from said transaction authorization device, wherein said data structure is responsive to said first information, said data structure comprises a random number, a first identification code, and third information, said third information comprises an encryption by a third encryption process of both second information and a signature responsive to said second information, a first portion of said second information is responsive to said first information, a second portion of said second information comprises said random number, a third portion of said second information comprises said first identification code, said random number is generated by said trusted authorization device, and said first identification code is associated with said trusted authorization device; and d. providing for communicating said data structure to a host server computer, wherein said data structure provides for a trusted authorization of said transaction.\",\n \"34.A method of providing for a trusted authorization of a transaction, comprising: a. providing for receiving by a first computer a data structure from a second computer, wherein said data structure is responsive to first information, said first information is related to a transaction to be authorized, said data structure comprises a random number, a first identification code, and third information, said third information comprises an encryption by a third encryption process of both second information and a signature by a first encryption process responsive to said second information, a first portion of said second information is responsive to said first information, a second portion of said second information comprises said random number, a third portion of said second information comprises said first identification code; b. providing for retrieving a set of stored working keys, wherein said operation of retrieving is responsive to said first identification code; c. providing for generating a set of session keys by a second encryption process, wherein said second encryption process is responsive to said random number and to said set of stored working keys; d. providing for generating second information and fifth information by decrypting said third information using said third encryption process that is responsive to said set of session keys; e. providing for generating a signature of said second information, wherein said signature is generated by said first encryption process; f. providing for comparing said signature with said fifth information; and g. if said signature matches said fifth information, then providing for acting upon said second information.\",\n \"35.A method of providing for a trusted authorization of a transaction as recited in claim 25, further comprising providing for transmitting to a third computer said random number, a set of encrypted working keys for said third encryption process and said third information, and receiving from said third computer a result, wherein said set of encrypted working keys is responsive to said first identification code, said set of encrypted working keys are encrypted with a set of keys of said third computer, the operation of providing for acting upon said second information is responsive to said result and said third computer performs the operations of providing for generating said set of session keys, providing for generating said second information and said fifth information, providing for generating said signature of said second information, and providing for comparing said signature with said fifth information.\",\n \"36.A method of authorizing a transaction responsive to a data structure, comprising: a. receiving said data structure, wherein said data structure is responsive to first information, said first information is related to a transaction to be authorized, said data structure comprises a random number, a first identification code, and third information, said third information comprises an encryption by a third encryption process of both second information and a signature by a first encryption process responsive to said second information, a first portion of said second information is responsive to said first information, a second portion of said second information comprises said random number, a third portion of said second information comprises said first identification code; b. retrieving or receiving a set of stored working keys, wherein said operation of retrieving is responsive to said first identification code; c. generating a set of session keys by a second encryption process, wherein said second encryption process is responsive to said random number and to said set of stored working keys; d. generating second information and fifth information by decrypting said third information using said third encryption process that is responsive to said set of session keys; e. generating a signature of said second information, wherein said signature is generated by said first encryption process; f. comparing said signature with said fifth information; and g. transmitting a result of the operation of comparing said signature with said fifth information.\",\n \"37.A method of authorizing a transaction responsive to a data structure as recited in claim 36, wherein said set of stored working keys are encrypted, further comprising decrypting said set of stored working keys prior to the operation of generating a set of session keys.\",\n \"38.A memory for storing data for access by an application program being executed on a computer, comprising a data structure stored in said memory, wherein said data structure is responsive to first information, said first information is related to a transaction to be authorized, and said data structure comprises a. a first data object comprising a random number; b. a second data object comprising a first identification code; and c. a third data object comprising third information, wherein said third information comprises an encryption by a third encryption process of both second information and a signature by a first encryption process responsive to said second information, said third encryption process is responsive to a set of session keys that are responsive to said random number, a first portion of said second information is responsive to said first information, a second portion of said second information comprises said random number, a third portion of said second information comprises said first identification code.\",\n \"39.A memory for storing data for access by an application program being executed on a computer as recited in claim 38, wherein said data structure further comprises a fourth data object comprising a value of a transaction counter.\",\n \"40.A memory for storing data for access by an application program being executed on a computer as recited in claim 38, wherein said second information incorporates a value of a transaction counter.\"\n ],\n [\n \"<SOH> BRIEF DESCRIPTION OF THE DRAWINGS <EOH>In the accompanying drawings: FIG.\",\n \"1 illustrates a block diagram of a trusted authorization device and an associated transaction processing system; FIG.\",\n \"2 illustrates a process for providing for a trusted authorization of a transaction from the point-of-view of an associated Trusted Authorization Device (TAD); FIG.\",\n \"3 illustrates as data structure used by a process for providing for a trusted authorization of a transaction; FIG.\",\n \"4 a illustrates an encryption process for generating a set of encryption keys; FIG.\",\n \"4 b illustrates a schematic representation of the process illustrated in FIG.\",\n \"4 a; FIG.\",\n \"5 a illustrates 3-DES encryption process for generating a set of encryption keys; FIG.\",\n \"5 b illustrates a schematic representation of the process illustrated in FIG.\",\n \"5 a; FIG.\",\n \"6 a illustrates 3-DES encryption process for encrypting a message; FIG.\",\n \"6 b illustrates a schematic representation of the process illustrated in FIG.\",\n \"6 a; FIG.\",\n \"7 a illustrates 3-DES decryption process for decrypting an encrypted message; FIG.\",\n \"7 b illustrates a schematic representation of the process illustrated in FIG.\",\n \"6 a; FIG.\",\n \"8 illustrates a process for providing for a trusted authorization of a transaction from the point-of-view of an associated client computer; FIG.\",\n \"9 illustrates a process for providing for a trusted authorization of a transaction from the point-of-view of an associated host computer; FIG.\",\n \"10 illustrates a process for providing for a trusted authorization of a transaction from the point-of-view of an associated Verification Decryption Server (VDS); FIG.\",\n \"11 a illustrates a key loading process from the point-of-view of an associated Key Loading Unit (KLU); FIG.\",\n \"11 b illustrates a schematic representation of the process illustrated in FIG.\",\n \"11 a; FIG.\",\n \"12 a illustrates a key loading process from the point-of-view of an associated Trusted Authorization Device; FIG.\",\n \"12 b illustrates a schematic representation of the process illustrated in FIG.\",\n \"11 a; FIG.\",\n \"13 illustrates a re-keying process from the point-of-view of an associated Key Loading Unit (KLU); FIG.\",\n \"14 illustrates a re-keying process from the point-of-view of an associated Trusted Authorization Device; FIG.\",\n \"15 illustrates a re-keying process from the point-of-view of an associated Trusted Authorization Device; FIG.\",\n \"16 illustrates a process for providing for a trusted authorization of a transaction from the point-of-view of an associated Trusted Authorization Device; FIG.\",\n \"17 illustrates a process for providing for a trusted authorization of a transaction from the point-of-view of an associated Verification Decryption Server (VDS); FIG.\",\n \"18 illustrates a table of TAD input commands; FIG.\",\n \"19 illustrates a table of TAD language codes; FIG.\",\n \"20 illustrates a structure of a TAD input command for authorizing data; FIGS.\",\n \"21 a illustrates a plaintext portion of a data structure of a TAD response to command for authorizing data; FIG.\",\n \"21 b illustrates an encrypted portion of a data structure of a TAD response to command for authorizing data; FIG.\",\n \"22 illustrates a table of data field types associated with a TAD data structure; FIG.\",\n \"23 illustrates a structure of an error response data packet from a TAD to a client computer if an authorization is either aborted or incorrect; FIG.\",\n \"24 illustrate a structure of TAD response to command for authorizing data; FIG.\",\n \"25 illustrate an example of a TAD data structure at various processing stages; FIG.\",\n \"26 a illustrates a structure of a TAD input command for loading a rekeying keyset; FIG.\",\n \"26 b illustrates a structure of a TAD response to a command for loading a rekeying keyset; FIG.\",\n \"27 a illustrates a structure of a TAD input command for installing a new working keyset; FIG.\",\n \"27 b illustrates a structure of a TAD response to a command for installing a new working keyset; FIG.\",\n \"28 a illustrates a structure of a TAD input command for installing a new language; FIG.\",\n \"28 b illustrates a structure of a TAD response to a command for installing a new language; FIG.\",\n \"29 a illustrates a structure of a TAD input command for identifying a TAD to a client computer; FIG.\",\n \"29 b illustrates a structure of a TAD response to a command for identifying a TAD to a client computer; FIG.\",\n \"30 a illustrates a structure of a TAD input command for testing a TAD maintenance key; FIG.\",\n \"30 b illustrates a structure of a TAD response to a command for testing a TAD maintenance key; FIG.\",\n \"31 a illustrates a structure of a TAD input command for personalizing a TAD; and FIG.\",\n \"31 b illustrates a structure of a TAD response to a personalizing a TAD.\",\n \"detailed-description description=\\\"Detailed Description\\\" end=\\\"lead\\\"?\"\n ],\n [\n \"BRIEF DESCRIPTION OF THE DRAWINGS In the accompanying drawings: FIG.\",\n \"1 illustrates a block diagram of a trusted authorization device and an associated transaction processing system; FIG.\",\n \"2 illustrates a process for providing for a trusted authorization of a transaction from the point-of-view of an associated Trusted Authorization Device (TAD); FIG.\",\n \"3 illustrates as data structure used by a process for providing for a trusted authorization of a transaction; FIG.\",\n \"4a illustrates an encryption process for generating a set of encryption keys; FIG.\",\n \"4b illustrates a schematic representation of the process illustrated in FIG.\",\n \"4a; FIG.\",\n \"5a illustrates 3-DES encryption process for generating a set of encryption keys; FIG.\",\n \"5b illustrates a schematic representation of the process illustrated in FIG.\",\n \"5a; FIG.\",\n \"6a illustrates 3-DES encryption process for encrypting a message; FIG.\",\n \"6b illustrates a schematic representation of the process illustrated in FIG.\",\n \"6a; FIG.\",\n \"7a illustrates 3-DES decryption process for decrypting an encrypted message; FIG.\",\n \"7b illustrates a schematic representation of the process illustrated in FIG.\",\n \"6a; FIG.\",\n \"8 illustrates a process for providing for a trusted authorization of a transaction from the point-of-view of an associated client computer; FIG.\",\n \"9 illustrates a process for providing for a trusted authorization of a transaction from the point-of-view of an associated host computer; FIG.\",\n \"10 illustrates a process for providing for a trusted authorization of a transaction from the point-of-view of an associated Verification Decryption Server (VDS); FIG.\",\n \"11a illustrates a key loading process from the point-of-view of an associated Key Loading Unit (KLU); FIG.\",\n \"11b illustrates a schematic representation of the process illustrated in FIG.\",\n \"11a; FIG.\",\n \"12a illustrates a key loading process from the point-of-view of an associated Trusted Authorization Device; FIG.\",\n \"12b illustrates a schematic representation of the process illustrated in FIG.\",\n \"11a; FIG.\",\n \"13 illustrates a re-keying process from the point-of-view of an associated Key Loading Unit (KLU); FIG.\",\n \"14 illustrates a re-keying process from the point-of-view of an associated Trusted Authorization Device; FIG.\",\n \"15 illustrates a re-keying process from the point-of-view of an associated Trusted Authorization Device; FIG.\",\n \"16 illustrates a process for providing for a trusted authorization of a transaction from the point-of-view of an associated Trusted Authorization Device; FIG.\",\n \"17 illustrates a process for providing for a trusted authorization of a transaction from the point-of-view of an associated Verification Decryption Server (VDS); FIG.\",\n \"18 illustrates a table of TAD input commands; FIG.\",\n \"19 illustrates a table of TAD language codes; FIG.\",\n \"20 illustrates a structure of a TAD input command for authorizing data; FIGS.\",\n \"21a illustrates a plaintext portion of a data structure of a TAD response to command for authorizing data; FIG.\",\n \"21b illustrates an encrypted portion of a data structure of a TAD response to command for authorizing data; FIG.\",\n \"22 illustrates a table of data field types associated with a TAD data structure; FIG.\",\n \"23 illustrates a structure of an error response data packet from a TAD to a client computer if an authorization is either aborted or incorrect; FIG.\",\n \"24 illustrate a structure of TAD response to command for authorizing data; FIG.\",\n \"25 illustrate an example of a TAD data structure at various processing stages; FIG.\",\n \"26a illustrates a structure of a TAD input command for loading a rekeying keyset; FIG.\",\n \"26b illustrates a structure of a TAD response to a command for loading a rekeying keyset; FIG.\",\n \"27a illustrates a structure of a TAD input command for installing a new working keyset; FIG.\",\n \"27b illustrates a structure of a TAD response to a command for installing a new working keyset; FIG.\",\n \"28a illustrates a structure of a TAD input command for installing a new language; FIG.\",\n \"28b illustrates a structure of a TAD response to a command for installing a new language; FIG.\",\n \"29a illustrates a structure of a TAD input command for identifying a TAD to a client computer; FIG.\",\n \"29b illustrates a structure of a TAD response to a command for identifying a TAD to a client computer; FIG.\",\n \"30a illustrates a structure of a TAD input command for testing a TAD maintenance key; FIG.\",\n \"30b illustrates a structure of a TAD response to a command for testing a TAD maintenance key; FIG.\",\n \"31a illustrates a structure of a TAD input command for personalizing a TAD; and FIG.\",\n \"31b illustrates a structure of a TAD response to a personalizing a TAD.\",\n \"DESCRIPTION OF EMBODIMENT(S) Electronic communications, and the data which traverses those communications, are relatively new, as is the technology used to protect electronic data.\",\n \"Existing communications protection technologies tend to fall into two categories.\",\n \"The first, government sponsored, is generally very well thought out and provides excellent protection, but is not readily available for commercial applications.\",\n \"The second, de facto commercial, are mostly not strong enough to protect important information, or are dedicated to specific functions.\",\n \"For example, standard point-of-sale devices are dedicated to merchandizing applications, and existing ATM systems are dedicated to the dispensing of cash.\",\n \"There exists a need for a device to provide personal protection of electronic data that is small, easy to use, provides excellent protection to the PC/laptop user, and that can operate in conjunction with corresponding devices at a central data gathering point to provide near real time validation of the information.\",\n \"As one example, involving financial transactions over the internet by a user, a financial institution may desire an enhanced level of security so as to verify that the user is who they say they are and that they have truly authorized a particular transaction.\",\n \"As another example, in a business-to-business environment, a paycheck processing company needs to know with virtual certainty the authenticity of instructions from associated business clients for making payroll distributions.\",\n \"As yet another example, in a gaming environment, a user of internet gaming services may wish to transfer funds from a credit card to a gaming card so as to participate in internet gaming, a transaction for which the credit card issuer generally demands authentication of the user and verification of the transaction so as to avoid a later repudiation of the transaction by the user.\",\n \"Referring to FIG.\",\n \"1, a Trusted Authorization Device 10 (or TAD) is operatively connected to an untrustworthy client 12 (for example, a personal computer or workstation PC) operated by a user 14.The TAD 10 provides a trustworthy subsystem that allows the authorization of electronic data transactions or actions while electronically connected to the untrustworthy client 12 platform in a potentially hostile environment.\",\n \"The TAD 10 comprises a trusted control processor 16, a trusted display 18, a trusted keypad 20 (e.g.\",\n \"a numeric keypad, or an alphanumeric keyboard), and a trusted device reader 22 (e.g.\",\n \"a magnetic stripe card, a chip card, a smart cart, etc.).\",\n \"In one embodiment, the TAD 10 is a computer ancillary hardware device that provides a trusted environment for the display and authorization of transactions/authorizations that can be compactly represented in human understandable form.\",\n \"The TAD 10 is operatively connected to the client 12 with a telecommunications channel 24, for example, a serial telecommunications (RS-232) interface that is constrained by the trusted control processor 16.A telecommunications channel 24 that is hardwired directly to the client 12 and not out-of-sight therefrom provides for enhanced security.\",\n \"The trusted control processor 16 and associated memory 26 are, for example, securely packaged in a tamper-proof, hardened housing, that if tampered with causes at least essential elements of the TAD 10 to either self-destruct or become inoperable and virtually undecipherable.\",\n \"For example, the trusted control processor 16 may be adapted with either a light sensor or a pressure sensor or both, which would cause the memory 26 to be erased response to an associated detection of light or pressure change that would result from tampering with the housing of the trusted control processor 16 and/or associated memory 26.The trusted control processor 16 is a dedicated CPU in the TAD 10 that controls and/or manages the trusted display 18, trusted keypad 20, trusted device reader 22, and the telecommunications channel 24.The trusted control processor 16 provides for the following capabilities: 1.A unique and well protected ID; 2.Embedded unique implementation of cryptographic algorithms (including digest algorithms, asymmetric encryption with public/private keys and symmetric encryption with private/private keys); 3.Random number generation; 4.Functionality to derive unique session keys per transaction from random generation sub process; 5.Cryptographic functionality to enable signing (digest and/or public/private encryption keys) and data encryption (private/private encryption keys) functionality; 6.Ability to protect the TAD encryption keys from observation or alteration.\",\n \"This involves on-chip hardening and system auto-destruct functionality in event of tamper detection.\",\n \"Tamper detection sensing is embedded in the TAD 10; 7.Ability to display proposed transactions supplied from host and load supplied information into appropriate data structures for transmission; 8.Ability to capture and load data from integrated TAD devices; 9.Client PC interface, trusted keypad, trusted device reader, e.g.\",\n \"card reader; 10.Optional camera, biometric input devices, and/or GPS receiver; and 11.Ability to integrate all authorized information into a transaction data structure, digitally sign this data structure, strongly and uniquely encrypt the appropriate portions of the structure, and transmit this data structure to the host for further transmission.\",\n \"The trusted display 18 is, for example, a separate display constrained by trusted control processor 16 and not subject to intercept, control or modification by a host system.\",\n \"The trusted display 18 displays the transaction or authorization to be performed, for example in a compact embodiment, on a 4 line by 20 character screen.\",\n \"The trusted keypad 20 is, for example, a numeric key pad (with alpha functionality) that is constrained by the trusted control processor 16 and whose data is not subject to interception, alteration, or replacement by signals from the client 12.The trusted keypad 20 is used by a user to enter information and accept or refuse a transaction.\",\n \"The trusted device reader 22 is, for example, a magnetic card/smart card reader that is constrained to communicate with the trusted control processor 16 and whose information is not subject to interception, alteration, or replacement by signals from the client 12.The trusted device reader 22 provides a means for the user 14 to provide proof of possession or the associated magnetic card/smart card, and thereby enable the TAD 10 to authenticate the transaction request.\",\n \"The trusted device reader 22 may, for example, be a hybrid card reader, enabling it to support the usage of chip cards in a trustworthy environment.\",\n \"Such chip cards provide an appropriate environment for accessing user-specific public key-enabled functionality.\",\n \"The client 12 is operatively connected, e.g.\",\n \"via the Internet, to a host server 28 having a communication interface 30.For example, the host server 28 could be operated by a service provider that requires an enhanced level of trust in the authorization of transactions or requests by the user 14 running particular application software of the service provider, and accordingly, who would provide a TAD 10 to the user 14 for authenticating transactions with the necessary enhanced level of trust.\",\n \"The host server 28 interfaces with a verification decryption server 32 (VDS), for example, via a VDS executive manager 34, and may also, or alternatively, interface with a customer application system 36 running associated application software, and also interfaced with the VDS 32.Each TAD 10 is provided with a unique alphanumeric ID (TADID_A) and a unique and well-protected binary ID (TADID_B), each of which are stored in memory 26.The alphanumeric ID (TADID_A) is also visible on the outside of an associated housing of the TAD 10 for purposes of identifying the particular device, for example for purposes of maintenance or physical distribution control.\",\n \"The associated trusted control processors 16 are also provided with embedded unique implementations of cryptographic algorithms, including at least one algorithm for generating a signature—which may include a digest process—(e.g.\",\n \"asymmetric encryption with public/private keys) and at least one algorithm for encrypting data (e.g.\",\n \"symmetric encryption with private/private keys), the later of which relies upon associated keys that are stored in the TAD 10 by a key loading unit 38 (KLU).\",\n \"Referring to FIG.\",\n \"2, in operation, in step (202) the untrustworthy client 12 downloads a proposed transaction or action to the TAD 10 for authorization, for example at the request of the user 14.Then, in step (204), the user 14, viewing the trusted display 18 and using the trusted keypad 20, pages through the proposed transaction, thereby providing a basis of trust for the action to be authorized.\",\n \"When the user 14 wishes to authorize the transaction, the user 14 can accept the transaction by pressing the appropriate key, or sequence of keys, on the trusted keypad 20.The user 14 may then further couple a unique physical token 40—e.g.\",\n \"a magnetic stripe card 40.1 or a smart card 40.2—to the trusted device reader 22, and then enter a personal PIN number associated therewith on the trusted keypad 20, thereby authenticating the identity of the user 14.Alternately, or in addition, the TAD 10 may require the user 14 to enter a PIN number associated with the TAD 10.Then, in step (206), if the user 14 authorizes the transaction, then the trusted control processor 16 stores the displayed first information, the captured card information, and all other necessary information (e.g.\",\n \"PIN, location from an associated trusted location device e.g.\",\n \"GPS receiver, etc) as second information in an associated data fields of an associated data structure 42, e.g.\",\n \"illustrated in FIG.\",\n \"3, wherein the second information is responsive to, or a function of the first information, and may comprise a copy of the first information.\",\n \"Otherwise, from step (206), the process repeats with step (202), wherein the TAD 10 awaits further input from the client 12.Then, in step (208), a random number generator 44—e.g.\",\n \"within the trusted control processor 16—generates a random number R, which may be either a pseudo-random number, or a true random number, for example, generated responsive to a noisy physical process.\",\n \"The TAD 10 may also access and increment a transaction counter, although this step is not essential.\",\n \"Then, in step (210), the trusted control processor 16 generates second information that is responsive to the first information displayed to the user, and which further incorporates the random number R and a first identification code of the TAD 10, e.g.\",\n \"the alphanumeric ID (TADID_A).\",\n \"Then, in step (212) the trusted control processor 16 generates a digital signature of the second information—thereby providing a basis for authorization and non-repudiation of the transaction—using a first encryption process, for example, an irreversible digest algorithm (e.g.\",\n \"an asymmetric encryption algorithm).\",\n \"Then, in steps (214) and (216), the trusted control processor 16 respectively retrieves a set of stored working keys Kw1, Kw2, Kw3 from the memory 26, and generates a set of transaction-specific session keys Ks1, Ks2, Ks3 using a second encryption process—generally illustrated in FIGS.\",\n \"4a, 4b, and illustrated for a 3-DES encryption process in FIGS.\",\n \"5a and 5b—using the random number R as a seed.\",\n \"The working keys Kw1, Kw2, Kw3 are stored in the memory 26 by a key loading process described hereinbelow.\",\n \"Then, in step (218), trusted control processor 16 generates third information by encrypting the combination of the second information from step (210), and the associated signature from step (212), using a third encryption process, for example a 3-DES encryption process as illustrated in FIGS.\",\n \"6a and 6b.\",\n \"Then, in step (220), the data structure 42 comprising the plaintext random number R, the plaintext alphanumeric ID (TADID_A), and the third information is communicated to the client 12, and communicated thereby to the host server 28.The host server 28 sends the data structure 42 to the VDS 32 for decryption and signature verification thereby, and if authenticated by the VDS 32, the transaction is processed by either the host server 28 or the associated customer application system 36.If a transaction counter is used, the value of the transaction counter would be incorporated in the second information (which is signed), and may also be incorporated as plaintext in the data structure 42.For example, if the user is interfaced with the host server 28 is via an Internet browser, a user may select, via the browser, a transaction to be conducted—for example, the transfer of funds from an account accessed via a standard financial card or a transaction within a custom domain.\",\n \"The browser then uses associated TAD 10 interface software to transfer the proposed transaction and instructions to the TAD 10 for authorization.\",\n \"Referring to FIG.\",\n \"8, from the point of view of the client 12, the above described process commences in step (802), wherein the user 14 initiates a transaction on a client 12 in communication with the host server 28.For example, the user 14 initiates a transaction on the Internet involving a purchase that the user 14 wishes to finance by a credit card, i.e.\",\n \"a magnetic stripe card 40.1.In step (804), responsive to the host server 28 requesting a trusted authorization of the transaction, a first information to be authorized is communicated to the TAD 10, and in step (806), if the user 14 has authorized the transaction using the TAD 10, the client 12 receives the associated data structure 42 from the TAD 10, and, in step (808), communicates this to the host server 28.Referring to FIG.\",\n \"9, from the point of view of the host server 28, in step (902), the host server 28 receives the data structure 42 from the client 12, in step (904), extracts the alphanumeric ID (TADID_A) from the plaintext portion of the data structure 42 and uses a lookup process to find an associated set of VDS encrypted TAD working keys KVDS—w1, KVDS—w2, KVDS—w3 that are encrypted using a VDS key that is not known by the host server 28.Then, in step (906), the VDS encrypted TAD working keys KVDS13 w1, KVDS—w2, KVDS—W3 and the data structure 42 are communicated by the host server 28 to the VDS 32, responsive to which, in step (908), the host server 28 receives a verification status from the VDS 32, and in step (910), the host server 28 communicates this verification status to the client 12.Referring to FIG.\",\n \"10, from the point of view of the verification decryption server 32 (VDS), in step (1002), the verification decryption server 32 receives the VDS encrypted TAD working keys KVDS—w1, KVDS—w2, KVDS—w3 and the data structure 42 from the host server 28.Then, in step (1004), the VDS 32 decrypts the VDS encrypted TAD working keys KVDS—w1, KVDS—w2, KVDS—w3 using associated VDS keys KVDS1, KVDS2, KVDS3 stored on the VDS 32 and loaded thereon by the key loading unit 38.Then, in step (1006), the VDS 32 extracts the plaintext random number R from the data structure 42, and in step (1008), uses the random number R as a seed, together with the decrypted TAD working keys Kw1, Kw2, Kw3 from step (1004) to generate—by a key generating process 500 as illustrated in FIGS.\",\n \"5a and 5b—a set of session keys Ks1, Ks2, Ks3 that are used in step (1010) to decrypt the encrypted portion (i.e.\",\n \"the second information) of the data structure 42, for example, in accordance with a 3-DES decryption process as illustrated in FIGS.\",\n \"7a and 7b.\",\n \"Then, in step (1012), the VDS 32 generates a signature of the second information using the same first encryption process as had been used in the TAD 10, and in step (1014), if the extracted, decrypted signature from step (1010) is the same as the generated signature from step (1012), then in step (1016), the verification process is successful, and in steps (1018) and (1020), the information responsive to the first information is extracted from the second information, and then communicated to the customer application system 36 to complete the transaction, after which the customer application system 36 notifies the host server 28 that the transaction has been completed successfully.\",\n \"Then, in step (1022), the verification status is communicated by the VDS 32 to the host server 28.If, from step (1014), the generated signature is not equal to the extracted, decrypted signature, then, in step (1024), the verification process is unsuccessful, and this verification status is communicated to the host server 28 in step (1022).\",\n \"The TAD 10 incorporates three sets of stored keys that are used in various encryption processes, 1) a set of read-only firmware keys KF1, KF2, KF3, 2) a set of rekeying keys KRK1, KRK2, KRK3 that are initially loaded on the TAD 10 by the key loading unit 38, and 3) a set of working keys Kw1, Kw2, Kw3 that are loaded on the TAD 10 during a rekeying operation by the key loading unit 38, either directly connected to the TAD 10, or remotely via a mailed floppy disk.\",\n \"The TAD 10 uses the working keys Kw1, Kw2, Kw3 to generate the transaction-specific session keys Ks1, Ks2, Ks3 in accordance with a key generating process 500, as described hereinabove.\",\n \"The key loading unit 38 loads the rekeying keys KRK1, KRK2, KRK3 and the working keys Kw1, Kw2, Kw3 on the TAD 10 by transferring an encrypted key seed to the TAD 10, after which the TAD 10 generates the respective keys using a key generating process 500, in which the TAD 10 and the key loading unit 38 each utilize prearranged secret keys to encrypt the key seed.\",\n \"For example, both binary ID (TADID_B) and the firmware keys KF1, KF2, KF3 are known to both the TAD 10 and the key loading unit 38, wherein the key loading unit 38 is able to determine the binary ID (TADID_B) from a lookup process, given the alphanumeric ID (TADID_A) of the TAD 10.Accordingly, both the key loading unit 38 and the TAD 10 can independently use the key generating process 500—with the binary ID (TADID_B) as the seed (with S1=S2=S3=TADID_B) and the firmware keys KF1, KF7, KF3 as the generating keys—to generate a set of maintenance keys maintenance keys KM1, KM2, KM3.The key loading unit 38 then uses the maintenance keys KM1, KM2, KM3 to encrypt a rekey random number RkR, which is then transferred in encrypted form to the TAD 10, which then decrypts the rekey random number RkR and uses this as a seed, together with the maintenance keys KM1, KM2, KM3 as generating keys, to generate the rekeying keys KRK1, KRK2, KRK3.Similarly, the key loading unit 38 can use the same rekey random number RkR as a seed, and the same maintenance keys KM1, KM2, KM3 as generating keys, to independently generate identical rekeying keys KRK1, KRK2, KRK3.Then, the key loading unit 38 can use the rekeying keys KRK1, KRK2, KRK3 to encrypt a working key random number that is transferred to the TAD 10 to be used thereby as a seed in accordance with the key generating process 500 to generate the TAD working keys Kw1, Kw2, Kw3, wherein the rekeying keys KRK1, KRK2, KRK3 are used as associated key generating keys.\",\n \"The key loading unit 38 is used to load the VDS encrypted TAD working keys KVDS—w1, KVDS—w2, KVDS—w3 on the VDS 32, using VDS keys for the encryption, after which the TAD rekeying keys KRK1, KRK2, KRK3 and TAD working keys Kw1, Kw2, Kw3 are destroyed on the key loading unit 38 so that the key loading unit 38 cannot become a single point source of security failure.\",\n \"The process by which the key loading unit 38 transfers and encrypted key seed to the TAD 10 is illustrated in FIG.\",\n \"11a, and is represented schematically in FIG.\",\n \"11b.\",\n \"The process by which the TAD 10 receives the encrypted key seed from the key loading unit 38 and decrypts the key seed is illustrated in FIG.\",\n \"12a, and is represented schematically in FIG.\",\n \"12b.\",\n \"The process by which respective key seeds for the rekeying keys KRK1, KRK2, KRK3 and the working keys Kw1, Kw2, Kw3 are respectively generated and encrypted by key loading unit 38, and transferred to the TAD 10, is illustrated in FIG.\",\n \"13.The process by which respective key seeds for the rekeying keys KRK1, KRK2, KRK3 and the working keys Kw1, Kw2, Kw3 are received by the TAD 10 and used to generate the respective rekeying keys KRK1, KRK2, KRK3 and working keys Kw1, Kw2, Kw3, is illustrated in FIG.\",\n \"14.Referring also to FIG.\",\n \"15, the key loading unit 38 provides for manual key distribution and management services using a limited set of commands.\",\n \"The TAD uses the maintenance key to decrypt the re-keying keys.\",\n \"This operation should be done locally via a direct connection to the key loading unit.\",\n \"The TAD relies upon a set of 3 DES re-keying keys to load the unit keys that the unit relies upon.\",\n \"These re-keying keys are installed in the TAD 10 by the key loading unit 38 via the TAD maintenance keys, which are internally generated in the TAD by the interaction of the TAD firmware keyset (which is common to all the TAD's in a production lot) with a TAD-specific 64 bit Binary ID, TADID_B.\",\n \"The three maintenance keys are generated by permuting the order of the firmware keyset in a triple DES EDE encryption of the BID, i.e.\",\n \"Km1=EKf1(DKf2(EKf3(TADID—B))), Km2=EKf3(DKf2(EKf1(TADID—B))), Km3=EKf2(DKf1(EKf3(TADID—B))), (wherein E and D respectively represent the encryption and decryption sub-processes of a symmetric encryption process, e.g.\",\n \"triple DES (3 DES).\",\n \"Kf1 is the first firmware key.\",\n \"Km1 is the first maintenance key.\",\n \"etc.\",\n \"Using the above processes the TAD can generate the TAD-specific maintenance keyset.\",\n \"Similarly, the keyloader, which knows the firmware keys and the BID can also generate and use the TAD-specific maintenance keyset.\",\n \"Thus, the keyloader can load the re-keying keys into the TAD.\",\n \"Using the maintenance keys Km1, Km2, Km3, the re-keying keys KRK1, KRK2, KRK3 are generated therein by execution of a load re-key command (a rekey command with no re-keying keys is assumed to be a load re-key command), having a command structure is: Clear text portion Re-key Command 4 bytes (0XFFFFFFFF) Re-keying block length 4 bytes (multiple of 8 bytes) Random number Rk 24 bytes Encrypted portion Random number length 4 Re-keying random number as specified (initially 24) digest prior to encryption as appropriate The size of the structures may vary with system versions.\",\n \"The encryption keys in process are the maintenance keys Km1, Km2, Km3.Upon receiving a re-key command with no re-keying keys, the TAD 10 performs the following functions: 1.Check to make sure that the block is the right length (if wrong, return a failure code and clear the block); 2.Calculate the session keys, using the random number Rk provided in the clear text portion, as follows: a.\",\n \"Take the first 8 bytes of the session_random number, R1_and calculate session key 1, Ks1=EKm1(DKm2(EKm3(R1))) (wherein E and D respectively represent the encryption and decryption sub-processes of a symmetric encryption process, e.g.\",\n \"triple DES (3 DES); b.\",\n \"Take the second 8 bytes of the session_random number, R2_and calculate session key 2, Ks2=EKm3(DKm2(EKm1(R2))); and c. Take the third 8 bytes of the session random number, R3 and calculate session key 3, Ks3=EKm2(DKm1(EKm3(R3))); 3.Decrypt the encrypted portion DKs3(Es2(DKs1(encrypted portion))) using CBC mode; 4.Calculate the MD5 hash of the entire block cleartext+decrypted portion (digest block set to NULL); 5.Compare calculated digest with received digest (if wrong, return a failure code and clear the block); 6.Take the first 8 bytes of the re-keying random number, RKR1 and calculate re-keying key 1, KrK1=EKm1(DKm2(EKm3(RkR1))); 7.Take the second 8 bytes of the re-keying random number, RkR2 and calculate re-keying key 2, KrK2=Ekm3(DKm2(EKm1(RkR2))); and 8.Take the third 8 bytes of the re-keying random number, RkR3 and calculate re-keying key 3, KrK3=EKm2(DKm1(EKm3(RkR3))).\",\n \"The re-keying command can be issued after the re-keying keys Kr1, Kr2, Kr3 have been installed in the TAD 10 by the key loading unit 38, wherein the key loading unit 38 calculates the re-keying keys Kr1, Kr2, Kr3 from the binary ID (TADID_B).\",\n \"After the re-keying keys Kr1, Kr2, Kr3 are installed in the TAD 10 by the key loading unit 38, the working keys Kw1, Kw2, Kw3 are generated therein by execution of a re-key command, having a command structure is: Plaintext portion Re-key Command 4 bytes (0XFFFFFFFF) Re-keying block length 4 bytes (multiple of 8 bytes) Random number Rk 24 bytes Encrypted portion Re-keying counter 4 Random number length 4 Re-keying random number as specified (initially 24) digest prior to encryption as appropriate The associated re-keying data structure, for example, does not have the flexibility of the general data structures.\",\n \"For example, the key loading unit 38 knows what algorithms are expected by the TAD 10 and uses the appropriate ones, hence there is no need for flexibility here.\",\n \"The size of the structures may vary with system versions.\",\n \"The encryption keys in process are not the working keys Kw1, Kw2, Kw3, but instead are the re-keying keys Kr1, Kr2, Kr3.Upon receiving a re-key command, the TAD 10 performs the following functions: 1.Check to make sure that the block is the right length (if wrong, return a failure code and clear the block); 2.Calculate the session keys, using the session random number Rk provided in the plaintext portion, as follows: a.\",\n \"Take the first 8 bytes of the session random number, R1, and calculate session key 1, Ks1: Ks1=EKr1(DKr2(Er3(R1))) (wherein E and D respectively represent the encryption and decryption sub-processes of a symmetric encryption process, e.g.\",\n \"triple DES (3 DES); b.\",\n \"Take the second 8 bytes of the session random number, R2, and calculate session key 2, Ks2: Ks2=EKr3(DKr2(EKr1(R2))); and c. Take the third 8 bytes of the session random number, R3 (alternately, R3 could be generated from R1 and R2 by R3=R1 XOR R2), and calculate session key 3, Ks3: Ks3=EKr2(DKr1(EKr3(R3))).\",\n \"3.Decrypt the encrypted portion DKs3(EKs2(DKs1(encrypted portion))) using CBC mode; 4.Calculate the MD5 hash of the entire block plaintext+decrypted portion (digest block set to NULL); 5.Compare calculated digest with received digest (if wrong, return a failure code and clear the block); 6.Check re-keying counter (which is initially set to 0).\",\n \"If less than current re-keying counter, return a failure code and clear the block; 7.Set re-keying counter and compute net unit working keys; 8.Take the first 8 bytes of the re-keying random number, RkR1 and calculate working key 1, Kw1: Kw1=EKr1(DKr2(EKr3(RkR1))); 9.Take the second 8 bytes of the re-keying random number, RkR2 and calculate working key 2, Kw2: Kw2=EKr3(DKr2(EKr1(RkR2))); and 10.Take the third 8 bytes of the re-keying random number, RkR3 (alternately, R3 could be generated from R1 and R2 by R3=R1 XOR R2), and calculate working key 3, Kw3: Kw3=EKr2(DKr1(EKr3(RkR3))).\",\n \"After the working keys Kw1, Kw2, Kw3 are initially loaded by the key loading unit 38, the TAD 10 is placed in service proximate to the client 12 for providing trusted signing and authorization of transactions.\",\n \"The one or more maintenance test keys are used for diagnosis and maintenance of the TAD 10, but generally not for purposes of data encryption.\",\n \"For example, in a diagnostic mode, the working keys are replaced with the maintenance keys, and are used to encode a dummy transaction, which can then be remotely decoded by maintenance personal to check that the TAD 10 is operating properly.\",\n \"The data encryption process utilizes a random number R generated by a random number generator 44 within the trusted control processor 16 as a seed for an irreversible digest process (e.g.\",\n \"an asymmetric encryption process), e.g.\",\n \"MD5, that signs a portion of token to be encrypted, and in combination with working keys Kw1, Kw2, Kw3 created by a separate key loading process and stored in memory 26, to generate a set of session keys Ks1, Ks2, Ks3 that are used in a symmetric encryption process, e.g.\",\n \"triple DES (3 DES) using cyclic block chaining (CBC) mode, to encrypt the signed message.\",\n \"The trusted control processor 16 also has a set of re-keying keys Kr1, Kr2, Kr3 that are generated by the key loading unit 38 in direct connection with the TAD 10 and stored in memory 26.The re-keying keys Kr1, Kr2, Kr3 thereafter enable the working keys Kw1, Kw2, Kw3 to be update remotely, for example, with new working keys Kw1, Kw2, Kw3 provided to the user 14 over a controlled path, e.g.\",\n \"via a floppy disc provided by mail or courier.\",\n \"Referring to FIGS.\",\n \"18, 19, 20, 26a, 27a, 28a, 29a, 30a, and 31a, the TAD 10 is controlled responsive to various types of TAD input commands that are communicated to the TAD 10 either by the client 12 or the key loading unit 38 over the telecommunications channel 24.FIGS.\",\n \"20 illustrates the syntax of a command by a client 12 to authorize the signing and encryption of data in accordance with the normal operation of the TAD 10.FIG.\",\n \"26a illustrates the syntax of a command by a client 12 or the key loading unit 38 to re-key the TAD 10.FIG.\",\n \"28a illustrates the syntax of a command by a client 12 or the key loading unit 38 to insert a new language in the TAD 10, by which messages are displayed on the trusted display 18.For example, the TAD 10 is configured for selectable predefined languages of English, French, German or Spanish, and is provided with the capability of loading other languages/character sets, e.g.\",\n \"of Asian languages.\",\n \"The TAD 10 has a straight forward interface with the client 12, for example, supporting the following three commands from the client: Identify—which returns a text string, the TAD ID#, and the TAD version number.\",\n \"Process_Transaction—which accepts a proposed transaction, if successful, returning a transaction packet Secure_Transaction—The TAD will display the commands to initiate the card swipe, transaction digest and encryption.\",\n \"The TAD command interface is as follows: Command 4 bytes Data length 4 bytes Data as specified If no data or result is present, the length is set to 0.Referring to FIG.\",\n \"16, the TAD 10 provides trusted authorization of transactions in accordance with the following process: 1.Client downloads transaction (a compact, <80 character string) to the TAD 10 in accordance with the command structure illustrated in FIG.\",\n \"5a.\",\n \"2.User observes displayed transaction and verifies that it is correct.\",\n \"a.\",\n \"If so, the user presses the GO/ACCEPT key.\",\n \"b.\",\n \"If not, the user presses the STOP/REJECT key.\",\n \"3.If rejected, the TAD 10 signals the client and returns a “reject” code.\",\n \"4.If accepted, the TAD 10 displays the downloaded instructions to the user.\",\n \"5.The user following the instruction, a. swipes/enters the specified card, b. enters their secret PIN and, c. presses the GO/ACCEPT key, d. Pressing the STOP/REJECT key clears the transaction.\",\n \"6.If rejected, the TAD 10 signals the client and returns a “reject” code.\",\n \"7.If the transaction was accepted, the TAD 10 a. digitally signs the captured card data and the PIN, b. packages this information with the transaction, c. appropriately encrypts the sensitive data, and d. prepares the transaction packet 8.The TAD 10 notifies the client of success and transfers the prepared transaction packet, or TAD Output Token, to the client, structured as indicated in FIGS.\",\n \"21a, 21b, and 24, and by example in FIG.\",\n \"25.The data is read in wire order.\",\n \"The TAD 10 is not limited to a particular type of encryption.\",\n \"The following table indicates an example of various algorithms that can be used for signing and encryption: Packet Digest Identifier 0D 3 DES CBC digest using packet working key set 1D MD5 2D MD160 3D SHA1 Packet Encryption Algorithm ID 0E 3 DES CBC using packet working key set 1E 1 DES using key 1 of packet working key set The client 12 then transmits this to the appropriate host server 28.The transaction generated by the TAD 10 will pass through the communications network and then received by a host server 28 at the customer's site.\",\n \"The host server 28 will contain the alphanumeric ID (TADID_A) identification number, and corresponding unique working keys Kw1, Kw2, Kw3 in an encrypted form.\",\n \"When the host server 28 receives a transaction from the client 12 operated by the user 14, the host uses the alphanumeric ID (TADID_A) to retrieve the associated encrypted working keys Kw1, Kw2, Kw3.The host server 28 passes the encrypted working keys Kw1, Kw2, Kw3 and the encrypted transaction to the verification decryption server 32, thereby operating in a stateless mode.\",\n \"The VDS 32 does not have to store or manage working keys Kw1, Kw2, Kw3 (although for performance reasons one would expect it to cache previously supplied keys).\",\n \"Referring to FIG.\",\n \"17, the VDS 32 initially operated as a single-threaded process operating entirely in memory, writing an audit log to hard disc.\",\n \"Once the VDS 32 accepts a transaction it completes the specified decryption and verification and returns the result before accepting another transaction.\",\n \"The host server 28 shall poll the VDS 32 before submitting a transaction for verification.\",\n \"The processes handled by the VDS 32, for example, includes the following: Read the supplied data block Read TAD ID Look up decryption key value Parse encrypted key data structure A VDS 32 has the decryption keys needed to decrypt the encrypted working keys Kw1, Kw2, Kw3.The host server 28 passes the encrypted working keys Kw1, Kw2, Kw3 to the VDS 32, with the following data format, for example, having a structure that is more general than that required to handle to initial implementation of 3 DES keys so as to be able to handle different secret or public key algorithms, as appropriate: Plaintext portion Element Identifier Element Length TAD Unit ID 8 bytes Key structure length 4 bytes Packet Digest Identifier 2 bytes Packet Encryption Algorithm ID 2 bytes Random number 16 bytes Encrypted portion Element Identifier Element Length TAD keys packet length − (32 + digest) digest prior to encryption as appropriate The duplication of databases may be prevented by storing the wrapped TAD keys with the TAD and user data in the host server 28.The use of two sets of unique DES key triplets: the unit re-keying key set, and the unit working key set, helps to reduce the processing and storage requirements that would otherwise be required with public key technology.\",\n \"The unit's re-keying key set is the basic key set for the unit.\",\n \"It can only be used with a single command, an encrypted command generated by the keying/re-keying server that causes the TAD 10 to generate the working keys Kw1, Kw2, Kw3 that are used in all security operations.\",\n \"The key loading unit 38 and host server 28 are trusted with the either the binary ID (TADID_B) or the working keys Kw1, Kw2, Kw3, an accordingly are, for example, implemented so as to provide substantial assurance of the integrity of the storage and processing of these keys, for example, based upon Getronics (formerly Wang) STOP platforms (NSA evaluated B3).\",\n \"The associated communications protection devices are examined by trusted third parties so as to provide independent assurance concerning the device properties and characteristics.\",\n \"The TAD 10 has been adapted to incorporate other devices into its trust perimeter such as a GPS receiver 46 and may be adapted to incorporate other devices, such as a signature input device 48, one or more biometric input devices 50 (e.g.\",\n \"voice, fingerprint, retinal scan), a camera, a breath analyzer 52, and etc.\",\n \"Referring to FIG.\",\n \"1, the security and authenticity of the TAD 10 may be with a GPS receiver 46, so as to enable a service provider to ascertain that the user 14 is at the correct prearranged physical location for the TAD 10, and that the TAD 10 has not been relocated.\",\n \"This is useful in certain business authorizations, where a TAD 10 is issued to a specific customer at a specific location.\",\n \"The use of the GPS receiver 48 allows the management system to provide strong assurance of the user's location, as well as provide an accurate time value (e.g.\",\n \"accurate to a millisecond or better).\",\n \"The coordinates (latitude, longitude, and/or altitude) from the GPS receiver 48 are stored in an associated field, e.g.\",\n \"Field #5 of the TAD Output data structure illustrated in FIG.\",\n \"24, and are therefore signed and encrypted along with the rest of the encrypted portion of the TAD Output data structure.\",\n \"Accordingly, the service provider include a test of the decrypted position coordinates of the TAD 10 from the GPS receiver 46 in deciding whether or not to carry out the transaction requested by the user 14.Whereas the TAD 10 is illustrated as a separate device, it should be understood that the TAD 10 could be embedded in the client 12.Furthermore, whereas the key loading unit 38 is illustrated as a single workstation within the trusted environment of the host server 28 and verification decryption server 32, it should be understood that the key loading unit 38 could be constructed as a portable unit that can be moved to the site of the TAD 10 by a trusted representative.\",\n \"While specific embodiments have been described in detail, those with ordinary skill in the art will appreciate that various modifications and alternatives to those details could be developed in light of the overall teachings of the disclosure.\",\n \"Accordingly, the particular arrangements disclosed are meant to be illustrative only and not limiting as to the scope of the invention, which is to be given the full breadth of the appended claims, and any and all equivalents thereof.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473842"}}},{"rowIdx":757,"cells":{"text":{"kind":"list like","value":[["Filter element, method for manufacture thereof, and filter using said element","The invention provides a filter element exhibiting an excellent deodorizing and adsorbing function with low pressure drop, as well as to a method for its manufacture and a filter using the element, and this is achieved by a filter element which satisfies the relationship that 0≦A≦1.1 mB+C and m=0.28−0.0005 D50, with the proviso that 0<C≦15, where D50 (μm) is the 50% particle diameter of the adsorbent powder, B (g/m2) is the amount of the adsorbent powder adhered to the base fabric, A (Pa) is the pressure drop of the filter as measured at room temperature under an air flow rate of 1 m/sec and C (Pa) is the pressure drop of the base fabric when coated with the adhesive and dried, as well as a method for its manufacture and a filter using the element.","The filter element is preferably manufactured by coating a base fabric with an adhesive and allowing the fabric to move in an electrostatic field formed by a high-voltage generating device to allow pulverized adsorbent powder with a specific mean particle size and standard deviation to electrostatically adhere to the base fabric surface, or by dispersing a pulverized adsorbent powder on the surface of a base fabric comprising heat-fusible fibers, and then partially fusing the heat-fusible fibers by heating to attach them to the adsorbent powder and allow the adsorbent powder to adhere to the base fabric surface."],["1.A filter element having pulverized adsorbent powder adhered to an adhesive-coated base fabric surface, which satisfies the relationship that 0≦A≦1.1 mB+C and m=0.28−0.0005 D50, with the proviso that 0<C≦15, where D50 (μm) is the 50% particle diameter of the adsorbent powder, B (g/m2) is the amount of the adsorbent powder adhered to the base fabric, A (Pa) is the pressure drop of the filter element as measured at room temperature under an air flow rate of 1 m/sec, and C (Pa) is the pressure drop of the base fabric coated with the adhesive and dried.","2.A filter element according to claim 1, wherein the 50% particle diameter D50 of said adsorbent powder is 10-1000 μm and the standard deviation σg for the particle size distribution of the adsorbent powder is 1.1-2.0.3.A filter element according to claim 1 or 2, wherein said adsorbent powder is adhered to both sides of the base fabric.","4.A filter element according to any one of claims 1 through 3, wherein said filter element is formed into a pleat form.","5.A filter element according to any one of claims 1 through 4, wherein said adsorbent powder is activated carbon powder.","6.A filter element according to any one of claims 1 through 5, wherein said base fabric is a nonwoven fabric employing heat-fusible fibers.","7.A filter element according to claim 6, wherein said nonwoven fabric is a nonwoven fabric composed of core-sheath fibers.","8.A filter element according to claim 7, wherein the core and sheath sections of said core-sheath fibers are composed of polyethylene terephthalate.","9.A filter element according to any one of claims 1 through 8, wherein said base fabric has a mass per unit area of 40-70 g/m2 and a thickness of 0.5-3 mm and contains fibers with a fineness of 10-20 dtex.","10.A filter element according to any one of claims 1 through 9, wherein protrusions are also formed in the base fabric.","11.A method for manufacture of a filter element by coating a base fabric with an adhesive and allowing the fabric to move in an electrostatic field formed by a high voltage generating device to allow pulverized adsorbent powder to electrostatically adhere to the base fabric surface.","12.A method for manufacture of a filter element by dispersing a pulverized adsorbent powder on the surface of a base fabric comprising a nonwoven fabric of heat-fusible fibers, and then partially fusing the heat-fusible fibers by heating to allow the adsorbent powder to adhere to the base fabric surface.","13.A filter comprising a combination of a cover sheet with a filter element according to any one of claims 1 through 10.14.A filter comprising a combination of an air filter and cover sheet with a filter element according to any one of claims 1 through 10.15.A filter according to claim 13 or 14, wherein the air filter and/or the cover sheet are nonwoven fabrics.","16.A filter according to claim 15, wherein said nonwoven fabric is a nonwoven fabric composed of core-sheath fibers.","17.A filter according to claim 16, wherein the core and sheath sections of said core-sheath fibers are composed of polyethylene terephthalate.","18.A filter according to any one of claims 13 through 17 which comprises a reinforcing material.","19.A filter comprising a combination of a material having a catalytic function with a filter according to any one of claims 13 through 17.20.A filter according to claim 19, wherein said material having a catalytic function is palladium chloride-impregnated activated carbon or a molded product thereof."],["<SOH> BACKGROUND ART <EOH>Activated carbon has been widely used in the past for noxious gas adsorption removal, gas purification and separation/recovery, gas occlusion, molecular sieve applications, and for decoloring purification in fields relating to foods or the chemical industry, water treatment, electric double layer condensers and the like, but activated carbon is also commonly used for various filters because of its excellent ability to adsorb and remove a variety of malodorous substances across a wide range.","Activated carbon for filters is often used in the form of powder or in pulverized form packed into a container, but it can also be conveniently used in many cases in the form of molded tubes or sheets.","In recent years, however, despite high expectations for such activated carbon filters, their uses have been limited because of a high degree of pressure drop.","Many types of filters modified to reduce pressure drop have been proposed to date, such as an adsorption filter comprising an adsorbent, a fine particle binder and reinforcing fibers, disclosed in Japanese Unexamined Patent Publication No.","HEI 3-151012.In this filter, the flat filter is made of particulate activated carbon coated with a thermoplastic material such as polyethylene, with packing in a frame set on a polypropylene net, while an example is given of stacking polyethylene nets for air purification.","Japanese Unexamined Patent Publication No.","HEI 3-238011 discloses an air purification filter comprising a laminated combination of an electret filter and a flat sheet.","This filter reduces pressure drop by employing a corrugated sheet.","Also, Japanese Unexamined Patent Publication No.","HEI 4-74505 discloses an air purification filter element with low pressure drop by laminating an electret filter and an adsorbent-containing filter into an integrally formed pleat.","However, these filters still exhibit high pressure drop of the flat sheet to which the activated carbon is attached, and hence their uses are often restricted.","On the other hand, there is disclosed in Japanese Unexamined Patent Publication No.","HEI 5-177133 an activated carbon-adhered fiber sheet wherein powdered activated carbon is adhered to fibers without an adhesive.","This sheet is obtained by dispersion, opening and activation of fibers such as glass fibers and a resin powder using an opening cylinder.","Also, Japanese Unexamined Patent Publication No.","HEI 6-219720 discloses activated carbon with a specified surface acidity, while Japanese Unexamined Patent Publication No.","HEI 9-271616 discloses a deodorizing filter material obtained by forming an activated carbon layer on one surface of a fiber sheet and forming an activated inorganic adsorbent layer on the other surface.","Also, U.S. Pat.","No.","5,124,177 and U.S. Pat.","No.","5,338,340 both disclose filters having activated carbon dispersed in an adhesive-coated sheet, where an air jet is sprayed to attach the activated carbon to the sheet.","However, although the techniques disclosed in these publications can provide firm attachment of the activated carbon to the sheet because they both employ pressing with a roll and high-pressure air blowing to attach the activated carbon to the sheet, it is still difficult to avoid a large pressure drop due to external pressing force on the activated carbon.","The present applicant had previously discovered that an easily fabricated flat filter with excellent adsorption performance can be produced by a fluidized adhesion method and has already filed Japanese Patent Publication No.","HEI 9-19920 as a patent application therefor (Japanese Unexamined Patent Publication No.","HEI 10-204384).","This method is a method of manufacturing a filter by attaching an adhesive to a sheet and passing the sheet through a fluidized bed of adsorbent particles, pressing at a pressure roll, shedding the insufficiently adhered particles and drying, and this may be said to be a highly efficient flat filter manufacturing method from the standpoint of production.","According to this method, however, the adsorbent such as activated carbon adheres in a randomly laminated fashion to the adhesive-coated sheet, and therefore the obtained filter does not always have low pressure drop.","In addition, although the insufficiently adhered particles are shed by vibration with a vibrator, the activated carbon adhered to the base fabric is sometimes shed during use, even after this shedding.","By utilizing a technique known as electrostatic setting, which is completely different from the manufacturing methods described above, it is possible to obtain an activated carbon sheet with low pressure drop and firm anchoring of activated carbon to the sheet.","As prior art adsorbent materials or deodorant materials comprising activated carbon fibers set in a sheet using the electrostatic setting technique, for example, Japanese Examined Patent Publication No.","SHO 57-31453 discloses an adsorbent material comprising activated carbon fibers set into a base material, and Japanese Unexamined Utility Model Publication No.","HEI 6-85034 discloses a deodorant material comprising activated carbon fibers electrostatically set on the surface of a sheet-like base material precoated with an adhesive on the back side.","Both of these methods employ activated carbon fibers as the material, but when the purpose of a filter is considered, since using activated carbon fibers means that a lower amount is used than when using particulate or pulverized activated carbon, the use of particulate or pulverized activated carbon is preferred from the standpoint of functional life.","As an example of the electrostatic setting technique being applied to produce an adsorbent layer using activated carbon as the material, Japanese Unexamined Patent Publication No.","SHO 50-144680 discloses a method of forming an adsorbent layer wherein an adsorbent element such as activated carbon is evenly adhered and anchored as an adsorbent layer by electrostatic adhesion.","Also, Japanese Unexamined Patent Publication No.","SHO 63-274429 describes a composite adhesive sheet wherein a sticking agent layer is provided on a porous sheet material and an adsorbent layer of activated carbon powder is provided on the sticking agent layer by electrostatic setting.","However, the adsorbent layer disclosed in Japanese Unexamined Patent Publication No.","SHO 50-144680 uses beady activated carbon, and being in the form of beads it does not always adhere satisfactorily to the sheet, such that a cost-effective product cannot be obtained.","The adsorbent sheet disclosed in Japanese Unexamined Patent Publication No.","SHO 63-274429 is used as a tobacco filter and therefore employs fine powdered activated carbon, with the binder being printed at a prescribed spacing in the porous sheet and the activated carbon powder being attached to the binder, and therefore the air permeability resistance is significant, as is also mentioned in the examples.","Fiber structures with high-performance permeability are also disclosed in Japanese Unexamined Patent Publication No.","HEI 10-102366, Japanese Patent Publication No.","2,818,693 and U.S. Pat.","No.","5,486,410.These comprise adsorbent particles of activated carbon or the like with heat-fusible fibers, and are described as having high air permeability.","However, the fiber-like structures disclosed in these publications have a structure in which the adsorbent particles are tightly held in the heat-fusible fibers, and are therefore described as having high air permeability, but considering the structure, a large pressure drop is unavoidable.","Low pressure drop is an important function required for filters, but despite numerous attempts to achieve it, no fully satisfactory solution has yet been found.","It is therefore an object of the present invention to provide a new filter element with firm adhesion of the adsorbent, without impairment of deodorizing function or adsorption performance, and with low pressure drop, as well as a method for its manufacture and a filter using the element."],["TECHNICAL FIELD The present invention relates to a filter element, to a method for its manufacture and to a filter using the element.","More specifically, the invention relates to a filter element having pulverized adsorbent powder attached to the surface of an adhesive-coated base fabric and exhibiting excellent deodorizing and adsorbing functions with low pressure drop, as well as to a method for its manufacture and a filter using the element.","Since the filter element of the invention has extremely low pressure drop relative to the amount of the activated carbon adhered to the base fabric, a filter employing the element can be suitably used as a filter for a variety of uses, including automobile cabin filters, indoor air purification filters and air conditioner filters.","BACKGROUND ART Activated carbon has been widely used in the past for noxious gas adsorption removal, gas purification and separation/recovery, gas occlusion, molecular sieve applications, and for decoloring purification in fields relating to foods or the chemical industry, water treatment, electric double layer condensers and the like, but activated carbon is also commonly used for various filters because of its excellent ability to adsorb and remove a variety of malodorous substances across a wide range.","Activated carbon for filters is often used in the form of powder or in pulverized form packed into a container, but it can also be conveniently used in many cases in the form of molded tubes or sheets.","In recent years, however, despite high expectations for such activated carbon filters, their uses have been limited because of a high degree of pressure drop.","Many types of filters modified to reduce pressure drop have been proposed to date, such as an adsorption filter comprising an adsorbent, a fine particle binder and reinforcing fibers, disclosed in Japanese Unexamined Patent Publication No.","HEI 3-151012.In this filter, the flat filter is made of particulate activated carbon coated with a thermoplastic material such as polyethylene, with packing in a frame set on a polypropylene net, while an example is given of stacking polyethylene nets for air purification.","Japanese Unexamined Patent Publication No.","HEI 3-238011 discloses an air purification filter comprising a laminated combination of an electret filter and a flat sheet.","This filter reduces pressure drop by employing a corrugated sheet.","Also, Japanese Unexamined Patent Publication No.","HEI 4-74505 discloses an air purification filter element with low pressure drop by laminating an electret filter and an adsorbent-containing filter into an integrally formed pleat.","However, these filters still exhibit high pressure drop of the flat sheet to which the activated carbon is attached, and hence their uses are often restricted.","On the other hand, there is disclosed in Japanese Unexamined Patent Publication No.","HEI 5-177133 an activated carbon-adhered fiber sheet wherein powdered activated carbon is adhered to fibers without an adhesive.","This sheet is obtained by dispersion, opening and activation of fibers such as glass fibers and a resin powder using an opening cylinder.","Also, Japanese Unexamined Patent Publication No.","HEI 6-219720 discloses activated carbon with a specified surface acidity, while Japanese Unexamined Patent Publication No.","HEI 9-271616 discloses a deodorizing filter material obtained by forming an activated carbon layer on one surface of a fiber sheet and forming an activated inorganic adsorbent layer on the other surface.","Also, U.S. Pat.","No.","5,124,177 and U.S. Pat.","No.","5,338,340 both disclose filters having activated carbon dispersed in an adhesive-coated sheet, where an air jet is sprayed to attach the activated carbon to the sheet.","However, although the techniques disclosed in these publications can provide firm attachment of the activated carbon to the sheet because they both employ pressing with a roll and high-pressure air blowing to attach the activated carbon to the sheet, it is still difficult to avoid a large pressure drop due to external pressing force on the activated carbon.","The present applicant had previously discovered that an easily fabricated flat filter with excellent adsorption performance can be produced by a fluidized adhesion method and has already filed Japanese Patent Publication No.","HEI 9-19920 as a patent application therefor (Japanese Unexamined Patent Publication No.","HEI 10-204384).","This method is a method of manufacturing a filter by attaching an adhesive to a sheet and passing the sheet through a fluidized bed of adsorbent particles, pressing at a pressure roll, shedding the insufficiently adhered particles and drying, and this may be said to be a highly efficient flat filter manufacturing method from the standpoint of production.","According to this method, however, the adsorbent such as activated carbon adheres in a randomly laminated fashion to the adhesive-coated sheet, and therefore the obtained filter does not always have low pressure drop.","In addition, although the insufficiently adhered particles are shed by vibration with a vibrator, the activated carbon adhered to the base fabric is sometimes shed during use, even after this shedding.","By utilizing a technique known as electrostatic setting, which is completely different from the manufacturing methods described above, it is possible to obtain an activated carbon sheet with low pressure drop and firm anchoring of activated carbon to the sheet.","As prior art adsorbent materials or deodorant materials comprising activated carbon fibers set in a sheet using the electrostatic setting technique, for example, Japanese Examined Patent Publication No.","SHO 57-31453 discloses an adsorbent material comprising activated carbon fibers set into a base material, and Japanese Unexamined Utility Model Publication No.","HEI 6-85034 discloses a deodorant material comprising activated carbon fibers electrostatically set on the surface of a sheet-like base material precoated with an adhesive on the back side.","Both of these methods employ activated carbon fibers as the material, but when the purpose of a filter is considered, since using activated carbon fibers means that a lower amount is used than when using particulate or pulverized activated carbon, the use of particulate or pulverized activated carbon is preferred from the standpoint of functional life.","As an example of the electrostatic setting technique being applied to produce an adsorbent layer using activated carbon as the material, Japanese Unexamined Patent Publication No.","SHO 50-144680 discloses a method of forming an adsorbent layer wherein an adsorbent element such as activated carbon is evenly adhered and anchored as an adsorbent layer by electrostatic adhesion.","Also, Japanese Unexamined Patent Publication No.","SHO 63-274429 describes a composite adhesive sheet wherein a sticking agent layer is provided on a porous sheet material and an adsorbent layer of activated carbon powder is provided on the sticking agent layer by electrostatic setting.","However, the adsorbent layer disclosed in Japanese Unexamined Patent Publication No.","SHO 50-144680 uses beady activated carbon, and being in the form of beads it does not always adhere satisfactorily to the sheet, such that a cost-effective product cannot be obtained.","The adsorbent sheet disclosed in Japanese Unexamined Patent Publication No.","SHO 63-274429 is used as a tobacco filter and therefore employs fine powdered activated carbon, with the binder being printed at a prescribed spacing in the porous sheet and the activated carbon powder being attached to the binder, and therefore the air permeability resistance is significant, as is also mentioned in the examples.","Fiber structures with high-performance permeability are also disclosed in Japanese Unexamined Patent Publication No.","HEI 10-102366, Japanese Patent Publication No.","2,818,693 and U.S. Pat.","No.","5,486,410.These comprise adsorbent particles of activated carbon or the like with heat-fusible fibers, and are described as having high air permeability.","However, the fiber-like structures disclosed in these publications have a structure in which the adsorbent particles are tightly held in the heat-fusible fibers, and are therefore described as having high air permeability, but considering the structure, a large pressure drop is unavoidable.","Low pressure drop is an important function required for filters, but despite numerous attempts to achieve it, no fully satisfactory solution has yet been found.","It is therefore an object of the present invention to provide a new filter element with firm adhesion of the adsorbent, without impairment of deodorizing function or adsorption performance, and with low pressure drop, as well as a method for its manufacture and a filter using the element.","DISCLOSURE OF THE INVENTION As a result of diligent research directed toward achieving the object stated above, the present inventors completed the invention upon finding that it is possible to obtain a filter element without reduced deodorizing function or adsorption performance and with low pressure drop by utilizing static electricity to attach a pulverized adsorbent having a specific mean particle size and a specific standard deviation for the particle size distribution, as a first method, or by dispersing a pulverized adsorbent powder on the surface of a base fabric composed of heat-fusible fibers and heating to partially fuse the heat-fusible fibers for attachment of the adsorbent powder to the base fabric surface, as a second method.","In other words, one aspect of the present invention provides a filter element having pulverized adsorbent powder adhered to an adhesive-coated base fabric, characterized in that the filter element satisfies the relationship that 0≦A≦1.1 mB+C and m=0.28−0.0005 D50, with the proviso that 0<C≦15, where D50 (μm) is the 50% particle diameter of the adsorbent powder, B (g/m2) is the amount of adhesion of the adsorbent to the base fabric, A (Pa) is the pressure drop of the filter as measured at room temperature under an air flow rate of 1 m/sec, and C (Pa) is the pressure drop of the base fabric when coated with the adhesive and dried.","Another aspect of the invention provides a method for manufacture of a filter element by coating a base fabric with an adhesive and allowing the fabric to move in an electrostatic field formed by a high voltage generating device to allow pulverized adsorbent powder to electrostatically adhere to the base fabric.","Still another aspect of the invention provides a method for manufacture of a filter element by dispersing a pulverized adsorbent powder on the surface of a base fabric comprising heat-fusible fibers, and then partially fusing the heat-fusible fibers by heating to allow the adsorbent powder to adhere to the base fabric surface.","Still another aspect of the invention provides a filter using the filter element described above.","BEST MODE FOR CARRYING OUT THE INVENTION As pulverized adsorbent powders to be used for the invention there may be mentioned activated carbon, activated alumina, activated clay, silica gel, zeolite or the like.","As activated carbon forms there may be mentioned coconut shell, palm coconut, fruit seed, sawdust, eucalyptus, pine or other plant-based, coal-based or petroleum-based coke materials and pitch carbides obtained from these starting materials, phenol-based resins, vinyl chloride-based resins, vinylidene chloride-based resins and vinyl alcohol-based resins.","The shape and size of the adsorbent are not particularly restricted, but the adsorbent will usually be pulverized, granular, fibrous or cylindrical with a size of about 10-1000 μm.","The base fabric for adhesion of the adsorbent is preferably one with air permeability, of which examples include nonwoven fabrics such as urethane foam, spun-bond and melt-blown nonwoven fabrics, dry nonwoven fabrics, wet nonwoven fabrics (paper), nonwoven fabrics made of polymers such as polyesters, polyamides, polypropylene and the like as the materials, as well as woven fabrics, cellulose and the like.","These may if desired be subjected to processing such as electret working or the like.","There are no particular restrictions on the adhesive coated onto the base fabric, and for example, a dispersion- or solution-type adhesive such as any of various latex- or acrylic-based emulsions, or an adhesive or tackifier composed of a synthetic resin such as a vinyl chloride-based resin, vinylidene chloride-based resin or acetic acid ester-based resin may be used.","Among these, acrylic-based emulsions are preferred.","The main feature of the filter element of the invention is that of extremely low pressure drop for the amount of adhesion of the adsorbent, compared to conventional filters.","According to the invention, the pressure drop of the filter element and of the dried base fabric after coating of the adhesive is measured using a device as prescribed by JIS B 9901-1997, at room temperature (normally 25° C.) under an air flow rate at 1 m/sec.","According to the invention, it is necessary to satisfy the relationship that 0≦A≦1.1 mB+C and m=0.28−0.0005 D50, with the proviso that 0<C≦15, and preferably that 0≦A≦1.05 mB+C and m=0.28−0.0005 D50, with the proviso that 0<C≦8, where D50 (μm) is the 50% particle diameter of the adsorbent powder, B (g/m2) is the amount of adhesion of the adsorbent powder to the base fabric, A (Pa) is the pressure drop of the filter element, and C (Pa) is the pressure drop of the base fabric coated with the adhesive and dried.","The amount of adhesion of the adsorbent powder according to the invention is represented in terms of the weight of the adsorbent attached per unit area of the base fabric.","In the filter element of the invention, the 50% particle diameter D50 of the adsorbent powder is 10-1000 μm and the standard deviation σg for the particle size distribution of the adsorbent powder is preferably 1.1-2.0, while the 50% particle diameter D50 is more preferably 200-600 μm.","In the filter element of the invention, the adsorbent powder is adhered to one or both sides of the base fabric surface, but adhesion to both sides is preferred from the standpoint of superior adsorption performance.","Because the filter element of the invention is used as a filter in combination with a cover sheet or the like, formation into a pleat form is preferred in practical terms.","Activated carbon is preferred as the adsorbent from the standpoint of achieving excellent deodorant performance and adsorption performance and superior adhesion to the base fabric.","The base fabric used may be heat-fusible fibers produced from a polyamide such as nylon, a polyester such as polyethylene terephthalate (PET) or polybutylene terephthalate (PBT), a modified polyester, or a polyolefin such as polyethylene or polypropylene.","The base fabric is preferably in the form of a nonwoven fabric, and the nonwoven fabric is preferably made of core-sheath fibers composed of heat-fusible fibers.","The core-sheath fibers preferably have a core section of PET, and a sheath section of modified PET.","The nonwoven fabric employing the heat-fusible fibers is preferably one with a mass per unit area of 40-70 g/m2, a thickness of 0.5-3 mm and containing fibers with a fineness of 10-20 dtex.","The fibers may also be mixed fibers so long as the fineness is 10-20 dtex.","The heat-fusible fibers may also be suitably used in a base fabric produced utilizing the principle of electrostatic setting.","The filter element of the invention is preferably manufactured utilizing the principle of electrostatic setting, or by heating and partially fusing heat-fusible fibers for attachment of the adsorbent powder to the base fabric surface, and when this method is employed, the adsorbent preferably has a specific mean particle size and a specific standard deviation, as mentioned above.","That is, according to the invention, the mean particle size of the adsorbent powder is 10-1000 μm and preferably 200-600 μm, and the standard deviation σg for the particle size distribution of the adsorbent powder, as defined by (15.87% particle size on a cumulative filter)/(50% particle size D50), where D50 (μm) is the 50% particle diameter, is preferably 1.1-2.0.Using an adsorbent with such a particle size distribution according to the invention can reduce the pressure drop of the filter element for the amount of adhesion of the adsorbent used, and provide excellent adsorption performance.","For production of a filter element of the invention utilizing the principle of electrostatic setting, first an electrostatic field is formed using a high voltage generating device.","For example, a high voltage may be applied to an electrode with a high voltage generating device to generate static electricity, and the base fabric may be passed through the electrostatic field at a prescribed distance along the electrode.","The moving speed of the base fabric and the distance between the base fabric and the electrode may be appropriately determined based on the size of the generated electrostatic field, the type of adsorbent used and the properties of the adhesive, but for most purposes the distance from the electrode may be 5-20 cm and the moving speed may be 1-10 m/min.","By providing an electrode on one side of the base fabric and running the adhesive-coated base fabric through the field, it is possible to adhere the adsorbent to the base fabric.","For adhesion of the adsorbent on both sides of the base fabric, the adhesive may be coated onto both sides of the base fabric and the procedure described above repeated twice, or else electrodes may be provided on both sides of the base fabric and the base fabric passed through the field for adhesion of the adsorbent to both sides simultaneously.","The base fabric may be transported horizontally through the electrostatic field, but moving the base fabric vertically up or down will tend to make the adsorbent adhered to the base fabric densely.","This is preferred for increasing the adsorption effect and reducing the pressure drop of the filter element.","Here, horizontal and vertical include slanted states and are not necessarily limited to horizontal and vertical in the strict sense.","Although the reason for the excellent effects of the filter element of the invention is not completely understood, it is thought that, for example, when the filter element is manufactured utilizing the principle of electrostatic setting, the randomly-shaped particles adhere densely in a single-layer fashion during adhesion of the pulverized adsorbent particles onto the base fabric in the electrostatic field, being oriented in the vertical direction relative to the base fabric.","Also, if the base fabric is moved downward or upward, the larger particles of the provided adsorbent fall further downward becoming the first to adhere, with the smaller particles adhering later, thereby allowing the smaller particles to be incorporated between the larger particles and resulting in the dense adhesion of the adsorbent particles to exhibit excellent adsorption performance.","For production of a filter element of the invention using heat-fusible fibers as the base fabric, the adsorbent powder may be evenly dispersed on the base fabric composed of the heat-fusible fibers, and then heating performed to partially fuse the heat-fusible fibers and adhere the adsorbent powder to the base fabric surface.","In this case, the adhesive may be used in combination with a heat-fusible adhesive such as polyethylene powder.","There are no particular restrictions on the heating means, and for example, it may be jet heating with heated air or irradiation with infrared rays.","The reason for which a filter element using heat-fusible fibers according to the invention exhibits low pressure drop without impairment of deodorizing function or adsorbing function is not fully understood, but it is believed that the low pressure drop without impairment of deodorizing function or adsorbing function is exhibited due to the low contact surface between the heat-fusible fibers and the adsorbent powder, and due to the bulky nonwoven fabric composition.","The filter element may also be fabricated using a base fabric wherein the heat-fusible fibers have been subjected to embossing to form protrusions on one or both sides.","When such a filter element is used in combination with a cover sheet, described below, on either the front or the back section, the cover sheet reliably bonds via the protrusions on the base fabric, resulting in a desirable situation of lower fusion bonding area and therefore low pressure drop.","The shapes, sizes and number of the protrusions is not restricted, and may be appropriately set as desired.","The filter element obtained in this manner is used as a filter in combination with a cover sheet, and the pressure drop is low even when used as a filter.","The filter element is preferably provided with cover sheets on both sides.","The filter element may also be suitably employed as a combination of an air filter and a cover sheet.","It is preferred to use a nonwoven fabric as the air filter and/or the cover sheet, and the nonwoven fabric used is preferably composed of the core-sheath type fibers described above.","PET is preferred as the core and sheath sections of the core-sheath fibers in the nonwoven fabric.","An electret-treated nonwoven fabric may also be used as the air filter.","The filter element preferably has a pleated structure, but for increased filter strength a metallic reinforcing material such as aluminum, iron, copper, titanium, stainless steel or the like, or a plastic reinforcing material such as polyethylene, polypropylene, polyester or the like may be inserted and set therein as appropriate.","Such reinforcing materials are preferably in the form of a comb, sheet or mesh.","The filter element of the invention may be used in a filter structure, but preferably its usefulness is extended by combining it with a material having a catalytic function.","Filters with composite functions are known, as described in Japanese Unexamined Patent Publication No.","HEI 9-271616, but no filters are known comprising materials with catalytic functions.","Materials with catalytic functions include, for example, activated carbon in combination with various metals such as copper, silver, gold, iron, manganese, platinum and palladium chloride, and their molded products may be used if necessary.","In particular, activated carbon impregnated with palladium chloride, or its molded product, may be used for removal of gases such as ethylene or carbon monoxide, to thereby notably increase the effect of the filter.","The material with a catalytic function may be provided at either the front or back part of the filter element, but for example, if an adsorption filter composed of activated carbon with a desulfurizing effect is set at the front part and a filter with a function of decomposing carbon monoxide, composed of palladium chloride or the like, is set at the back part, for treatment of noxious gas, the life of the back filter with the catalytic function can be extended and it is preferable.","Various combinations of filter elements of the invention and materials with catalytic functions may of course be used.","The invention will now be explained in greater detail through the following examples.","EXAMPLE 1 A PET nonwoven fabric (SKC50 by Hikotomi Industrial Co., Ltd., mass per unit area: 50 g/m2, pressure drop of nonwoven fabric alone: 6 Pa) was used as the base fabric, and an acrylic-based emulsion (Nipol LX820A acryl emulsion, solid portion: 22 g/m2, product of Nihon Zeon Co., Ltd.) was evenly coated onto both sides of the base fabric with a gravure coater (Asahi Precision Roll Lattice Model 35).","A voltage of 23 kV was applied using an electrostatic tester by Kasuga Electric Works Ltd. to produce an electrostatic field.","The electrode spacing distance was 10 cm, and the adhesive-coated base fabric (30 cm×1 m) was passed through horizontally at a speed of 1 m/min.","GG26/60 35N (D50: 430 μm, σg: 1.2) activated carbon by Kuraray Chemical Co., Ltd. was supplied at the same time, and the activated carbon was adhered thereto.","The adhesion of the activated carbon onto the base fabric, after removal of the moisture in the adhesive using a roller drier, was 250 g/m2 on both sides of the base fabric.","The pressure drop of the filter element was measured to be 21 Pa by passing air through at 1 m/sec at 25° C. The pressure drop of the dried base fabric after coating of the adhesive was 7.5 Pa.","The relationship between pressure drop and amount of the activated carbon powder adhered to the base fabric satisfied the relationship that 0≦A≦1.05 mB+C and m=0.28−0.0005D50, where 0<C≦8.The adsorptions (%) of toluene, sulfur dioxide and n-butane after 1 minute of gas flow were measured according to JIS B 9901-1997, as indices of the adsorption performance.","The results are shown in Table 1.EXAMPLE 2 A filter element was manufactured in the same manner as Example 1, except that the activated carbon was adhered only to one side of the base fabric.","The amount of the activated carbon powder adhered to the base fabric was 130 g/m2, the pressure drop of the filter element was 12 Pa, the pressure drop of the base fabric after coating and drying the adhesive was 7 Pa.","The filter element satisfied the relationship that 0≦A≦1.05 mB+C and m=0.28−0.0005 D50, where 0<C≦8, was satisfied.","The results of measurement of the adsorption performance are shown in Table 1.COMPARATIVE EXAMPLE 1 The same base fabric, adhesive and activated carbon used in Example 1 were used to manufacture a filter element by the fluidized adhesion method under the same conditions as indicated in Example 1 of Japanese Unexamined Patent Publication No.","HEI 10-204384.As shown by the results in Table 1 including the adsorption performance, the pressure drop was large at 40 Pa. TABLE 1 Adsorption (%) Pressure Sulfur drop (Pa) Toluene dioxide n-Butane Example 1 21 95 96 91 Example 2 12 82 77 61 Comparative 40 96 96 92 Example 1 EXAMPLE 3 A filter element was manufactured in the same manner as Example 1, except that a heat-fusible fiber of nonwoven fabric (OEX by Kurashiki Textile Manufacturing Co., Ltd., fiber size: 17 dtex, mass perunit area: 64 g/m2, thickness: 1.6 mm, pressure drop of nonwoven fabric alone: 3 Pa) was used as the base fabric.","The amount of the activated carbon adhered to the base fabric was 330 g/m2.The filter element had a pressure drop of 17.5 Pa when measured by passing 25° C. air at 1 m/sec.","The pressure drop of the base fabric after coating and drying the adhesive was 4.5 Pa.","The relationship between the pressure drop and the amount of the activated carbon adhered to the base fabric satisfied the relationship that 0≦A≦1.05 mB+C and m=0.28−0.0005 D50, where 0<C≦8.EXAMPLE 4 AND COMPARATIVE EXAMPLES 2-3 A nonwoven fabric (Spun Bond PK102 by Mitsui Chemical Co., Ltd., basis weight: 13 g/m2) was provided on both sides of the filter element manufactured in Example 1, and the pressure drop and adsorption were measured.","Two automobile filters of the same type by Japan Vilene Co., Ltd. mounted in commercial automobiles were also measured for pressure drop and adsorption.","The results are shown in Table 2.The filter of Example 4 was clearly superior in terms of pressure drop and adsorption performance.","The filter element was also worked into a pleated shape and cut, reinforced with a comb-shaped polypropylene reinforcing material at several locations on the back of the filter element and provided with a nonwoven fabric on the front for use as an air conditioner filter.","Prolonged stable operation was possible, and the performance as a filter was satisfactorily adequate.","EXAMPLE 5 AND COMPARATIVE EXAMPLE 4 A nonwoven fabric (Spun Bond PK102 by Mitsui Chemical Co., Ltd., mass per unit area: 13 g/m2) was provided on both sides of the filter element manufactured in Example 1, and after further providing an electret nonwoven fabric for air filtration, the pressure drop and adsorption were measured.","An automobile filter of the same type by Japan Vilene Co., Ltd. mounted in a commercial automobile was also measured for pressure drop and adsorption.","The results are shown in Table 2.The filter of Example 5 was clearly superior in terms of pressure drop and adsorption performance.","EXAMPLE 6 AND COMPARATIVE EXAMPLE 5 Activated carbon was adhered to only one side of a base fabric, and a filter element was manufactured by the same process as in Example 1.A nonwoven fabric was provided on both sides and the pressure drop and the adsorption were measured in the same manner as Example 4.For comparison, a commercial automobile-mounted automobile filter by Freudenberg Co. composed of the relatively low amount of the activated carbon adhered to the base fabric was also measured for pressure drop and adsorption (Comparative Example 5).","The results are shown in Table 2.The filter of Example 6 was clearly superior in terms of pressure drop and adsorption performance.","TABLE 2 Amount of the activated carbon adhered Pres- Total to the sure Adsorption (%) weight base fabric drop Sulfur n- (g/m2) (g/m2) (Pa) Toluene dioxide Butane Example 4 370 270 43.0 96.5 98.0 94.1 Comparative 373 293 82.4 92.4 95.7 73.4 Example 2 Comparative 491 417 104.0 96.4 94.5 73.3 Example 3 Example 5 390 270 78.0 96.5 98.0 94.1 Comparative 356 247 109.0 92.5 93.1 67.5 Example 4 Example 6 189 141 22.0 78.8 74.8 70.3 Comparative 512 249 43.0 86.0 86.5 54.0 Example 5 COMPARATIVE EXAMPLE 6 A filter element was manufactured in the same manner as Example 1, except that a heat-fusible fiber of nonwoven fabric (COLBACK50, CD50 by ACORDIS Co., fineness: 10-15 dtex, mass per unit area: 50 g/m2, thickness: 0.34 mm, pressure drop of nonwoven fabric alone: 6.3 Pa) was used as the base fabric.","The amount of the activated carbon adhered to the base fabric was 210 g/m2, the pressure drop was 40 Pa, the pressure drop of the base fabric was 8 Pa after coating and drying the adhesive.","When the thickness of the nonwoven fabric was too thin, high pressure drop resulted, relationship that 0≦A≦1.1 mB+C and m=0.28−0.0005 D50, where 0<C≦15 was not satisfied.","EXAMPLE 7 An emboss roll set to 120° C. was used to form regular hexagonal depressions bound by a bank width of 0.2 mm, a bank height of 0.7 mm, a bank side length of 2.0 mm and a bank angle (base angle indicating base of trapezoidal cross-section) of 600 in the heat-fusible fiber of nonwoven fabric used in Example 3, on both sides of the base fabric in a turtle-shell fashion, and a filter element was manufactured in the same manner as Example 3 except for using this nonwoven fabric as the base fabric.","The pressure drop measured in the same manner was 25 Pa, and the pressure drop of the filter manufactured in the same manner as Example 4 was measured to be 45 Pa.","EXAMPLE 8 A filter element was manufactured by the same method using the same nonwoven fabric as in Example 3, except that activated carbon (GG26/60 35N) by Kuraray Chemical Co., Ltd. was adhered at 390 g/m2 to the top side and palladium chloride-impregnated carbon (T-E32/60) by Kuraray Chemical Co., Ltd. was adhered at 345 g/m2 to the bottom side.","The filter element was cut to 400 mm×500 mm and housed in a 1 m3 test box.","Air containing 50 ppm SO2 and 200 ppm CO was circulated through the filter element for 5 hours at a linear speed of 1 m/sec.","This was repeated for 5 cycles, and the residual gas was analyzed.","For comparison, there were prepared filter elements with activated carbon (GG26/60 35N) by Kuraray Chemical Co., Ltd. adhered at 390 g/m2 only to the top side of the filter element (Reference Example 1) and with palladium chloride-impregnated carbon (T-E32/60) by Kuraray Chemical Co., Ltd. adhered at 345 g/m2 only to the bottom side of the filter element (Reference Example 2), air containing 50 ppm SO2 and 200 ppm CO was circulated through the filter element for 5 hours at a linear speed of 1 m/sec in the same manner as before for 5 cycles, and the residual gas was analyzed.","Absolutely no SO2 was detected in the residual gas with either of the filter elements.","Also, compared to 100 as the CO residue rate for Reference Example 1, the CO residue rate was 80 for Reference Example 2 and 50 when activated carbon and palladium chloride-impregnated carbon were adhered, thus clearly showing an effect by the filter element with the activated carbon and palladium chloride-impregnated carbon adhered thereto.","Industrial Applicability According to the present invention there is provided a filter element exhibiting an excellent deodorizing function and adsorbing performance with low pressure drop, as well as a method for its manufacture and a filter using the element.","Since the filter of the invention has an excellent deodorizing function and adsorbing performance as well as low pressure drop, as mentioned above, it can be suitably used as a filter for various uses, including automobile cabin filters, indoor air purification filters and air conditioner filters, which require high deodorizing and adsorption effects with low pressure drop."]],"string":"[\n [\n \"Filter element, method for manufacture thereof, and filter using said element\",\n \"The invention provides a filter element exhibiting an excellent deodorizing and adsorbing function with low pressure drop, as well as to a method for its manufacture and a filter using the element, and this is achieved by a filter element which satisfies the relationship that 0≦A≦1.1 mB+C and m=0.28−0.0005 D50, with the proviso that 0<C≦15, where D50 (μm) is the 50% particle diameter of the adsorbent powder, B (g/m2) is the amount of the adsorbent powder adhered to the base fabric, A (Pa) is the pressure drop of the filter as measured at room temperature under an air flow rate of 1 m/sec and C (Pa) is the pressure drop of the base fabric when coated with the adhesive and dried, as well as a method for its manufacture and a filter using the element.\",\n \"The filter element is preferably manufactured by coating a base fabric with an adhesive and allowing the fabric to move in an electrostatic field formed by a high-voltage generating device to allow pulverized adsorbent powder with a specific mean particle size and standard deviation to electrostatically adhere to the base fabric surface, or by dispersing a pulverized adsorbent powder on the surface of a base fabric comprising heat-fusible fibers, and then partially fusing the heat-fusible fibers by heating to attach them to the adsorbent powder and allow the adsorbent powder to adhere to the base fabric surface.\"\n ],\n [\n \"1.A filter element having pulverized adsorbent powder adhered to an adhesive-coated base fabric surface, which satisfies the relationship that 0≦A≦1.1 mB+C and m=0.28−0.0005 D50, with the proviso that 0<C≦15, where D50 (μm) is the 50% particle diameter of the adsorbent powder, B (g/m2) is the amount of the adsorbent powder adhered to the base fabric, A (Pa) is the pressure drop of the filter element as measured at room temperature under an air flow rate of 1 m/sec, and C (Pa) is the pressure drop of the base fabric coated with the adhesive and dried.\",\n \"2.A filter element according to claim 1, wherein the 50% particle diameter D50 of said adsorbent powder is 10-1000 μm and the standard deviation σg for the particle size distribution of the adsorbent powder is 1.1-2.0.3.A filter element according to claim 1 or 2, wherein said adsorbent powder is adhered to both sides of the base fabric.\",\n \"4.A filter element according to any one of claims 1 through 3, wherein said filter element is formed into a pleat form.\",\n \"5.A filter element according to any one of claims 1 through 4, wherein said adsorbent powder is activated carbon powder.\",\n \"6.A filter element according to any one of claims 1 through 5, wherein said base fabric is a nonwoven fabric employing heat-fusible fibers.\",\n \"7.A filter element according to claim 6, wherein said nonwoven fabric is a nonwoven fabric composed of core-sheath fibers.\",\n \"8.A filter element according to claim 7, wherein the core and sheath sections of said core-sheath fibers are composed of polyethylene terephthalate.\",\n \"9.A filter element according to any one of claims 1 through 8, wherein said base fabric has a mass per unit area of 40-70 g/m2 and a thickness of 0.5-3 mm and contains fibers with a fineness of 10-20 dtex.\",\n \"10.A filter element according to any one of claims 1 through 9, wherein protrusions are also formed in the base fabric.\",\n \"11.A method for manufacture of a filter element by coating a base fabric with an adhesive and allowing the fabric to move in an electrostatic field formed by a high voltage generating device to allow pulverized adsorbent powder to electrostatically adhere to the base fabric surface.\",\n \"12.A method for manufacture of a filter element by dispersing a pulverized adsorbent powder on the surface of a base fabric comprising a nonwoven fabric of heat-fusible fibers, and then partially fusing the heat-fusible fibers by heating to allow the adsorbent powder to adhere to the base fabric surface.\",\n \"13.A filter comprising a combination of a cover sheet with a filter element according to any one of claims 1 through 10.14.A filter comprising a combination of an air filter and cover sheet with a filter element according to any one of claims 1 through 10.15.A filter according to claim 13 or 14, wherein the air filter and/or the cover sheet are nonwoven fabrics.\",\n \"16.A filter according to claim 15, wherein said nonwoven fabric is a nonwoven fabric composed of core-sheath fibers.\",\n \"17.A filter according to claim 16, wherein the core and sheath sections of said core-sheath fibers are composed of polyethylene terephthalate.\",\n \"18.A filter according to any one of claims 13 through 17 which comprises a reinforcing material.\",\n \"19.A filter comprising a combination of a material having a catalytic function with a filter according to any one of claims 13 through 17.20.A filter according to claim 19, wherein said material having a catalytic function is palladium chloride-impregnated activated carbon or a molded product thereof.\"\n ],\n [\n \"<SOH> BACKGROUND ART <EOH>Activated carbon has been widely used in the past for noxious gas adsorption removal, gas purification and separation/recovery, gas occlusion, molecular sieve applications, and for decoloring purification in fields relating to foods or the chemical industry, water treatment, electric double layer condensers and the like, but activated carbon is also commonly used for various filters because of its excellent ability to adsorb and remove a variety of malodorous substances across a wide range.\",\n \"Activated carbon for filters is often used in the form of powder or in pulverized form packed into a container, but it can also be conveniently used in many cases in the form of molded tubes or sheets.\",\n \"In recent years, however, despite high expectations for such activated carbon filters, their uses have been limited because of a high degree of pressure drop.\",\n \"Many types of filters modified to reduce pressure drop have been proposed to date, such as an adsorption filter comprising an adsorbent, a fine particle binder and reinforcing fibers, disclosed in Japanese Unexamined Patent Publication No.\",\n \"HEI 3-151012.In this filter, the flat filter is made of particulate activated carbon coated with a thermoplastic material such as polyethylene, with packing in a frame set on a polypropylene net, while an example is given of stacking polyethylene nets for air purification.\",\n \"Japanese Unexamined Patent Publication No.\",\n \"HEI 3-238011 discloses an air purification filter comprising a laminated combination of an electret filter and a flat sheet.\",\n \"This filter reduces pressure drop by employing a corrugated sheet.\",\n \"Also, Japanese Unexamined Patent Publication No.\",\n \"HEI 4-74505 discloses an air purification filter element with low pressure drop by laminating an electret filter and an adsorbent-containing filter into an integrally formed pleat.\",\n \"However, these filters still exhibit high pressure drop of the flat sheet to which the activated carbon is attached, and hence their uses are often restricted.\",\n \"On the other hand, there is disclosed in Japanese Unexamined Patent Publication No.\",\n \"HEI 5-177133 an activated carbon-adhered fiber sheet wherein powdered activated carbon is adhered to fibers without an adhesive.\",\n \"This sheet is obtained by dispersion, opening and activation of fibers such as glass fibers and a resin powder using an opening cylinder.\",\n \"Also, Japanese Unexamined Patent Publication No.\",\n \"HEI 6-219720 discloses activated carbon with a specified surface acidity, while Japanese Unexamined Patent Publication No.\",\n \"HEI 9-271616 discloses a deodorizing filter material obtained by forming an activated carbon layer on one surface of a fiber sheet and forming an activated inorganic adsorbent layer on the other surface.\",\n \"Also, U.S. Pat.\",\n \"No.\",\n \"5,124,177 and U.S. Pat.\",\n \"No.\",\n \"5,338,340 both disclose filters having activated carbon dispersed in an adhesive-coated sheet, where an air jet is sprayed to attach the activated carbon to the sheet.\",\n \"However, although the techniques disclosed in these publications can provide firm attachment of the activated carbon to the sheet because they both employ pressing with a roll and high-pressure air blowing to attach the activated carbon to the sheet, it is still difficult to avoid a large pressure drop due to external pressing force on the activated carbon.\",\n \"The present applicant had previously discovered that an easily fabricated flat filter with excellent adsorption performance can be produced by a fluidized adhesion method and has already filed Japanese Patent Publication No.\",\n \"HEI 9-19920 as a patent application therefor (Japanese Unexamined Patent Publication No.\",\n \"HEI 10-204384).\",\n \"This method is a method of manufacturing a filter by attaching an adhesive to a sheet and passing the sheet through a fluidized bed of adsorbent particles, pressing at a pressure roll, shedding the insufficiently adhered particles and drying, and this may be said to be a highly efficient flat filter manufacturing method from the standpoint of production.\",\n \"According to this method, however, the adsorbent such as activated carbon adheres in a randomly laminated fashion to the adhesive-coated sheet, and therefore the obtained filter does not always have low pressure drop.\",\n \"In addition, although the insufficiently adhered particles are shed by vibration with a vibrator, the activated carbon adhered to the base fabric is sometimes shed during use, even after this shedding.\",\n \"By utilizing a technique known as electrostatic setting, which is completely different from the manufacturing methods described above, it is possible to obtain an activated carbon sheet with low pressure drop and firm anchoring of activated carbon to the sheet.\",\n \"As prior art adsorbent materials or deodorant materials comprising activated carbon fibers set in a sheet using the electrostatic setting technique, for example, Japanese Examined Patent Publication No.\",\n \"SHO 57-31453 discloses an adsorbent material comprising activated carbon fibers set into a base material, and Japanese Unexamined Utility Model Publication No.\",\n \"HEI 6-85034 discloses a deodorant material comprising activated carbon fibers electrostatically set on the surface of a sheet-like base material precoated with an adhesive on the back side.\",\n \"Both of these methods employ activated carbon fibers as the material, but when the purpose of a filter is considered, since using activated carbon fibers means that a lower amount is used than when using particulate or pulverized activated carbon, the use of particulate or pulverized activated carbon is preferred from the standpoint of functional life.\",\n \"As an example of the electrostatic setting technique being applied to produce an adsorbent layer using activated carbon as the material, Japanese Unexamined Patent Publication No.\",\n \"SHO 50-144680 discloses a method of forming an adsorbent layer wherein an adsorbent element such as activated carbon is evenly adhered and anchored as an adsorbent layer by electrostatic adhesion.\",\n \"Also, Japanese Unexamined Patent Publication No.\",\n \"SHO 63-274429 describes a composite adhesive sheet wherein a sticking agent layer is provided on a porous sheet material and an adsorbent layer of activated carbon powder is provided on the sticking agent layer by electrostatic setting.\",\n \"However, the adsorbent layer disclosed in Japanese Unexamined Patent Publication No.\",\n \"SHO 50-144680 uses beady activated carbon, and being in the form of beads it does not always adhere satisfactorily to the sheet, such that a cost-effective product cannot be obtained.\",\n \"The adsorbent sheet disclosed in Japanese Unexamined Patent Publication No.\",\n \"SHO 63-274429 is used as a tobacco filter and therefore employs fine powdered activated carbon, with the binder being printed at a prescribed spacing in the porous sheet and the activated carbon powder being attached to the binder, and therefore the air permeability resistance is significant, as is also mentioned in the examples.\",\n \"Fiber structures with high-performance permeability are also disclosed in Japanese Unexamined Patent Publication No.\",\n \"HEI 10-102366, Japanese Patent Publication No.\",\n \"2,818,693 and U.S. Pat.\",\n \"No.\",\n \"5,486,410.These comprise adsorbent particles of activated carbon or the like with heat-fusible fibers, and are described as having high air permeability.\",\n \"However, the fiber-like structures disclosed in these publications have a structure in which the adsorbent particles are tightly held in the heat-fusible fibers, and are therefore described as having high air permeability, but considering the structure, a large pressure drop is unavoidable.\",\n \"Low pressure drop is an important function required for filters, but despite numerous attempts to achieve it, no fully satisfactory solution has yet been found.\",\n \"It is therefore an object of the present invention to provide a new filter element with firm adhesion of the adsorbent, without impairment of deodorizing function or adsorption performance, and with low pressure drop, as well as a method for its manufacture and a filter using the element.\"\n ],\n [\n \"TECHNICAL FIELD The present invention relates to a filter element, to a method for its manufacture and to a filter using the element.\",\n \"More specifically, the invention relates to a filter element having pulverized adsorbent powder attached to the surface of an adhesive-coated base fabric and exhibiting excellent deodorizing and adsorbing functions with low pressure drop, as well as to a method for its manufacture and a filter using the element.\",\n \"Since the filter element of the invention has extremely low pressure drop relative to the amount of the activated carbon adhered to the base fabric, a filter employing the element can be suitably used as a filter for a variety of uses, including automobile cabin filters, indoor air purification filters and air conditioner filters.\",\n \"BACKGROUND ART Activated carbon has been widely used in the past for noxious gas adsorption removal, gas purification and separation/recovery, gas occlusion, molecular sieve applications, and for decoloring purification in fields relating to foods or the chemical industry, water treatment, electric double layer condensers and the like, but activated carbon is also commonly used for various filters because of its excellent ability to adsorb and remove a variety of malodorous substances across a wide range.\",\n \"Activated carbon for filters is often used in the form of powder or in pulverized form packed into a container, but it can also be conveniently used in many cases in the form of molded tubes or sheets.\",\n \"In recent years, however, despite high expectations for such activated carbon filters, their uses have been limited because of a high degree of pressure drop.\",\n \"Many types of filters modified to reduce pressure drop have been proposed to date, such as an adsorption filter comprising an adsorbent, a fine particle binder and reinforcing fibers, disclosed in Japanese Unexamined Patent Publication No.\",\n \"HEI 3-151012.In this filter, the flat filter is made of particulate activated carbon coated with a thermoplastic material such as polyethylene, with packing in a frame set on a polypropylene net, while an example is given of stacking polyethylene nets for air purification.\",\n \"Japanese Unexamined Patent Publication No.\",\n \"HEI 3-238011 discloses an air purification filter comprising a laminated combination of an electret filter and a flat sheet.\",\n \"This filter reduces pressure drop by employing a corrugated sheet.\",\n \"Also, Japanese Unexamined Patent Publication No.\",\n \"HEI 4-74505 discloses an air purification filter element with low pressure drop by laminating an electret filter and an adsorbent-containing filter into an integrally formed pleat.\",\n \"However, these filters still exhibit high pressure drop of the flat sheet to which the activated carbon is attached, and hence their uses are often restricted.\",\n \"On the other hand, there is disclosed in Japanese Unexamined Patent Publication No.\",\n \"HEI 5-177133 an activated carbon-adhered fiber sheet wherein powdered activated carbon is adhered to fibers without an adhesive.\",\n \"This sheet is obtained by dispersion, opening and activation of fibers such as glass fibers and a resin powder using an opening cylinder.\",\n \"Also, Japanese Unexamined Patent Publication No.\",\n \"HEI 6-219720 discloses activated carbon with a specified surface acidity, while Japanese Unexamined Patent Publication No.\",\n \"HEI 9-271616 discloses a deodorizing filter material obtained by forming an activated carbon layer on one surface of a fiber sheet and forming an activated inorganic adsorbent layer on the other surface.\",\n \"Also, U.S. Pat.\",\n \"No.\",\n \"5,124,177 and U.S. Pat.\",\n \"No.\",\n \"5,338,340 both disclose filters having activated carbon dispersed in an adhesive-coated sheet, where an air jet is sprayed to attach the activated carbon to the sheet.\",\n \"However, although the techniques disclosed in these publications can provide firm attachment of the activated carbon to the sheet because they both employ pressing with a roll and high-pressure air blowing to attach the activated carbon to the sheet, it is still difficult to avoid a large pressure drop due to external pressing force on the activated carbon.\",\n \"The present applicant had previously discovered that an easily fabricated flat filter with excellent adsorption performance can be produced by a fluidized adhesion method and has already filed Japanese Patent Publication No.\",\n \"HEI 9-19920 as a patent application therefor (Japanese Unexamined Patent Publication No.\",\n \"HEI 10-204384).\",\n \"This method is a method of manufacturing a filter by attaching an adhesive to a sheet and passing the sheet through a fluidized bed of adsorbent particles, pressing at a pressure roll, shedding the insufficiently adhered particles and drying, and this may be said to be a highly efficient flat filter manufacturing method from the standpoint of production.\",\n \"According to this method, however, the adsorbent such as activated carbon adheres in a randomly laminated fashion to the adhesive-coated sheet, and therefore the obtained filter does not always have low pressure drop.\",\n \"In addition, although the insufficiently adhered particles are shed by vibration with a vibrator, the activated carbon adhered to the base fabric is sometimes shed during use, even after this shedding.\",\n \"By utilizing a technique known as electrostatic setting, which is completely different from the manufacturing methods described above, it is possible to obtain an activated carbon sheet with low pressure drop and firm anchoring of activated carbon to the sheet.\",\n \"As prior art adsorbent materials or deodorant materials comprising activated carbon fibers set in a sheet using the electrostatic setting technique, for example, Japanese Examined Patent Publication No.\",\n \"SHO 57-31453 discloses an adsorbent material comprising activated carbon fibers set into a base material, and Japanese Unexamined Utility Model Publication No.\",\n \"HEI 6-85034 discloses a deodorant material comprising activated carbon fibers electrostatically set on the surface of a sheet-like base material precoated with an adhesive on the back side.\",\n \"Both of these methods employ activated carbon fibers as the material, but when the purpose of a filter is considered, since using activated carbon fibers means that a lower amount is used than when using particulate or pulverized activated carbon, the use of particulate or pulverized activated carbon is preferred from the standpoint of functional life.\",\n \"As an example of the electrostatic setting technique being applied to produce an adsorbent layer using activated carbon as the material, Japanese Unexamined Patent Publication No.\",\n \"SHO 50-144680 discloses a method of forming an adsorbent layer wherein an adsorbent element such as activated carbon is evenly adhered and anchored as an adsorbent layer by electrostatic adhesion.\",\n \"Also, Japanese Unexamined Patent Publication No.\",\n \"SHO 63-274429 describes a composite adhesive sheet wherein a sticking agent layer is provided on a porous sheet material and an adsorbent layer of activated carbon powder is provided on the sticking agent layer by electrostatic setting.\",\n \"However, the adsorbent layer disclosed in Japanese Unexamined Patent Publication No.\",\n \"SHO 50-144680 uses beady activated carbon, and being in the form of beads it does not always adhere satisfactorily to the sheet, such that a cost-effective product cannot be obtained.\",\n \"The adsorbent sheet disclosed in Japanese Unexamined Patent Publication No.\",\n \"SHO 63-274429 is used as a tobacco filter and therefore employs fine powdered activated carbon, with the binder being printed at a prescribed spacing in the porous sheet and the activated carbon powder being attached to the binder, and therefore the air permeability resistance is significant, as is also mentioned in the examples.\",\n \"Fiber structures with high-performance permeability are also disclosed in Japanese Unexamined Patent Publication No.\",\n \"HEI 10-102366, Japanese Patent Publication No.\",\n \"2,818,693 and U.S. Pat.\",\n \"No.\",\n \"5,486,410.These comprise adsorbent particles of activated carbon or the like with heat-fusible fibers, and are described as having high air permeability.\",\n \"However, the fiber-like structures disclosed in these publications have a structure in which the adsorbent particles are tightly held in the heat-fusible fibers, and are therefore described as having high air permeability, but considering the structure, a large pressure drop is unavoidable.\",\n \"Low pressure drop is an important function required for filters, but despite numerous attempts to achieve it, no fully satisfactory solution has yet been found.\",\n \"It is therefore an object of the present invention to provide a new filter element with firm adhesion of the adsorbent, without impairment of deodorizing function or adsorption performance, and with low pressure drop, as well as a method for its manufacture and a filter using the element.\",\n \"DISCLOSURE OF THE INVENTION As a result of diligent research directed toward achieving the object stated above, the present inventors completed the invention upon finding that it is possible to obtain a filter element without reduced deodorizing function or adsorption performance and with low pressure drop by utilizing static electricity to attach a pulverized adsorbent having a specific mean particle size and a specific standard deviation for the particle size distribution, as a first method, or by dispersing a pulverized adsorbent powder on the surface of a base fabric composed of heat-fusible fibers and heating to partially fuse the heat-fusible fibers for attachment of the adsorbent powder to the base fabric surface, as a second method.\",\n \"In other words, one aspect of the present invention provides a filter element having pulverized adsorbent powder adhered to an adhesive-coated base fabric, characterized in that the filter element satisfies the relationship that 0≦A≦1.1 mB+C and m=0.28−0.0005 D50, with the proviso that 0<C≦15, where D50 (μm) is the 50% particle diameter of the adsorbent powder, B (g/m2) is the amount of adhesion of the adsorbent to the base fabric, A (Pa) is the pressure drop of the filter as measured at room temperature under an air flow rate of 1 m/sec, and C (Pa) is the pressure drop of the base fabric when coated with the adhesive and dried.\",\n \"Another aspect of the invention provides a method for manufacture of a filter element by coating a base fabric with an adhesive and allowing the fabric to move in an electrostatic field formed by a high voltage generating device to allow pulverized adsorbent powder to electrostatically adhere to the base fabric.\",\n \"Still another aspect of the invention provides a method for manufacture of a filter element by dispersing a pulverized adsorbent powder on the surface of a base fabric comprising heat-fusible fibers, and then partially fusing the heat-fusible fibers by heating to allow the adsorbent powder to adhere to the base fabric surface.\",\n \"Still another aspect of the invention provides a filter using the filter element described above.\",\n \"BEST MODE FOR CARRYING OUT THE INVENTION As pulverized adsorbent powders to be used for the invention there may be mentioned activated carbon, activated alumina, activated clay, silica gel, zeolite or the like.\",\n \"As activated carbon forms there may be mentioned coconut shell, palm coconut, fruit seed, sawdust, eucalyptus, pine or other plant-based, coal-based or petroleum-based coke materials and pitch carbides obtained from these starting materials, phenol-based resins, vinyl chloride-based resins, vinylidene chloride-based resins and vinyl alcohol-based resins.\",\n \"The shape and size of the adsorbent are not particularly restricted, but the adsorbent will usually be pulverized, granular, fibrous or cylindrical with a size of about 10-1000 μm.\",\n \"The base fabric for adhesion of the adsorbent is preferably one with air permeability, of which examples include nonwoven fabrics such as urethane foam, spun-bond and melt-blown nonwoven fabrics, dry nonwoven fabrics, wet nonwoven fabrics (paper), nonwoven fabrics made of polymers such as polyesters, polyamides, polypropylene and the like as the materials, as well as woven fabrics, cellulose and the like.\",\n \"These may if desired be subjected to processing such as electret working or the like.\",\n \"There are no particular restrictions on the adhesive coated onto the base fabric, and for example, a dispersion- or solution-type adhesive such as any of various latex- or acrylic-based emulsions, or an adhesive or tackifier composed of a synthetic resin such as a vinyl chloride-based resin, vinylidene chloride-based resin or acetic acid ester-based resin may be used.\",\n \"Among these, acrylic-based emulsions are preferred.\",\n \"The main feature of the filter element of the invention is that of extremely low pressure drop for the amount of adhesion of the adsorbent, compared to conventional filters.\",\n \"According to the invention, the pressure drop of the filter element and of the dried base fabric after coating of the adhesive is measured using a device as prescribed by JIS B 9901-1997, at room temperature (normally 25° C.) under an air flow rate at 1 m/sec.\",\n \"According to the invention, it is necessary to satisfy the relationship that 0≦A≦1.1 mB+C and m=0.28−0.0005 D50, with the proviso that 0<C≦15, and preferably that 0≦A≦1.05 mB+C and m=0.28−0.0005 D50, with the proviso that 0<C≦8, where D50 (μm) is the 50% particle diameter of the adsorbent powder, B (g/m2) is the amount of adhesion of the adsorbent powder to the base fabric, A (Pa) is the pressure drop of the filter element, and C (Pa) is the pressure drop of the base fabric coated with the adhesive and dried.\",\n \"The amount of adhesion of the adsorbent powder according to the invention is represented in terms of the weight of the adsorbent attached per unit area of the base fabric.\",\n \"In the filter element of the invention, the 50% particle diameter D50 of the adsorbent powder is 10-1000 μm and the standard deviation σg for the particle size distribution of the adsorbent powder is preferably 1.1-2.0, while the 50% particle diameter D50 is more preferably 200-600 μm.\",\n \"In the filter element of the invention, the adsorbent powder is adhered to one or both sides of the base fabric surface, but adhesion to both sides is preferred from the standpoint of superior adsorption performance.\",\n \"Because the filter element of the invention is used as a filter in combination with a cover sheet or the like, formation into a pleat form is preferred in practical terms.\",\n \"Activated carbon is preferred as the adsorbent from the standpoint of achieving excellent deodorant performance and adsorption performance and superior adhesion to the base fabric.\",\n \"The base fabric used may be heat-fusible fibers produced from a polyamide such as nylon, a polyester such as polyethylene terephthalate (PET) or polybutylene terephthalate (PBT), a modified polyester, or a polyolefin such as polyethylene or polypropylene.\",\n \"The base fabric is preferably in the form of a nonwoven fabric, and the nonwoven fabric is preferably made of core-sheath fibers composed of heat-fusible fibers.\",\n \"The core-sheath fibers preferably have a core section of PET, and a sheath section of modified PET.\",\n \"The nonwoven fabric employing the heat-fusible fibers is preferably one with a mass per unit area of 40-70 g/m2, a thickness of 0.5-3 mm and containing fibers with a fineness of 10-20 dtex.\",\n \"The fibers may also be mixed fibers so long as the fineness is 10-20 dtex.\",\n \"The heat-fusible fibers may also be suitably used in a base fabric produced utilizing the principle of electrostatic setting.\",\n \"The filter element of the invention is preferably manufactured utilizing the principle of electrostatic setting, or by heating and partially fusing heat-fusible fibers for attachment of the adsorbent powder to the base fabric surface, and when this method is employed, the adsorbent preferably has a specific mean particle size and a specific standard deviation, as mentioned above.\",\n \"That is, according to the invention, the mean particle size of the adsorbent powder is 10-1000 μm and preferably 200-600 μm, and the standard deviation σg for the particle size distribution of the adsorbent powder, as defined by (15.87% particle size on a cumulative filter)/(50% particle size D50), where D50 (μm) is the 50% particle diameter, is preferably 1.1-2.0.Using an adsorbent with such a particle size distribution according to the invention can reduce the pressure drop of the filter element for the amount of adhesion of the adsorbent used, and provide excellent adsorption performance.\",\n \"For production of a filter element of the invention utilizing the principle of electrostatic setting, first an electrostatic field is formed using a high voltage generating device.\",\n \"For example, a high voltage may be applied to an electrode with a high voltage generating device to generate static electricity, and the base fabric may be passed through the electrostatic field at a prescribed distance along the electrode.\",\n \"The moving speed of the base fabric and the distance between the base fabric and the electrode may be appropriately determined based on the size of the generated electrostatic field, the type of adsorbent used and the properties of the adhesive, but for most purposes the distance from the electrode may be 5-20 cm and the moving speed may be 1-10 m/min.\",\n \"By providing an electrode on one side of the base fabric and running the adhesive-coated base fabric through the field, it is possible to adhere the adsorbent to the base fabric.\",\n \"For adhesion of the adsorbent on both sides of the base fabric, the adhesive may be coated onto both sides of the base fabric and the procedure described above repeated twice, or else electrodes may be provided on both sides of the base fabric and the base fabric passed through the field for adhesion of the adsorbent to both sides simultaneously.\",\n \"The base fabric may be transported horizontally through the electrostatic field, but moving the base fabric vertically up or down will tend to make the adsorbent adhered to the base fabric densely.\",\n \"This is preferred for increasing the adsorption effect and reducing the pressure drop of the filter element.\",\n \"Here, horizontal and vertical include slanted states and are not necessarily limited to horizontal and vertical in the strict sense.\",\n \"Although the reason for the excellent effects of the filter element of the invention is not completely understood, it is thought that, for example, when the filter element is manufactured utilizing the principle of electrostatic setting, the randomly-shaped particles adhere densely in a single-layer fashion during adhesion of the pulverized adsorbent particles onto the base fabric in the electrostatic field, being oriented in the vertical direction relative to the base fabric.\",\n \"Also, if the base fabric is moved downward or upward, the larger particles of the provided adsorbent fall further downward becoming the first to adhere, with the smaller particles adhering later, thereby allowing the smaller particles to be incorporated between the larger particles and resulting in the dense adhesion of the adsorbent particles to exhibit excellent adsorption performance.\",\n \"For production of a filter element of the invention using heat-fusible fibers as the base fabric, the adsorbent powder may be evenly dispersed on the base fabric composed of the heat-fusible fibers, and then heating performed to partially fuse the heat-fusible fibers and adhere the adsorbent powder to the base fabric surface.\",\n \"In this case, the adhesive may be used in combination with a heat-fusible adhesive such as polyethylene powder.\",\n \"There are no particular restrictions on the heating means, and for example, it may be jet heating with heated air or irradiation with infrared rays.\",\n \"The reason for which a filter element using heat-fusible fibers according to the invention exhibits low pressure drop without impairment of deodorizing function or adsorbing function is not fully understood, but it is believed that the low pressure drop without impairment of deodorizing function or adsorbing function is exhibited due to the low contact surface between the heat-fusible fibers and the adsorbent powder, and due to the bulky nonwoven fabric composition.\",\n \"The filter element may also be fabricated using a base fabric wherein the heat-fusible fibers have been subjected to embossing to form protrusions on one or both sides.\",\n \"When such a filter element is used in combination with a cover sheet, described below, on either the front or the back section, the cover sheet reliably bonds via the protrusions on the base fabric, resulting in a desirable situation of lower fusion bonding area and therefore low pressure drop.\",\n \"The shapes, sizes and number of the protrusions is not restricted, and may be appropriately set as desired.\",\n \"The filter element obtained in this manner is used as a filter in combination with a cover sheet, and the pressure drop is low even when used as a filter.\",\n \"The filter element is preferably provided with cover sheets on both sides.\",\n \"The filter element may also be suitably employed as a combination of an air filter and a cover sheet.\",\n \"It is preferred to use a nonwoven fabric as the air filter and/or the cover sheet, and the nonwoven fabric used is preferably composed of the core-sheath type fibers described above.\",\n \"PET is preferred as the core and sheath sections of the core-sheath fibers in the nonwoven fabric.\",\n \"An electret-treated nonwoven fabric may also be used as the air filter.\",\n \"The filter element preferably has a pleated structure, but for increased filter strength a metallic reinforcing material such as aluminum, iron, copper, titanium, stainless steel or the like, or a plastic reinforcing material such as polyethylene, polypropylene, polyester or the like may be inserted and set therein as appropriate.\",\n \"Such reinforcing materials are preferably in the form of a comb, sheet or mesh.\",\n \"The filter element of the invention may be used in a filter structure, but preferably its usefulness is extended by combining it with a material having a catalytic function.\",\n \"Filters with composite functions are known, as described in Japanese Unexamined Patent Publication No.\",\n \"HEI 9-271616, but no filters are known comprising materials with catalytic functions.\",\n \"Materials with catalytic functions include, for example, activated carbon in combination with various metals such as copper, silver, gold, iron, manganese, platinum and palladium chloride, and their molded products may be used if necessary.\",\n \"In particular, activated carbon impregnated with palladium chloride, or its molded product, may be used for removal of gases such as ethylene or carbon monoxide, to thereby notably increase the effect of the filter.\",\n \"The material with a catalytic function may be provided at either the front or back part of the filter element, but for example, if an adsorption filter composed of activated carbon with a desulfurizing effect is set at the front part and a filter with a function of decomposing carbon monoxide, composed of palladium chloride or the like, is set at the back part, for treatment of noxious gas, the life of the back filter with the catalytic function can be extended and it is preferable.\",\n \"Various combinations of filter elements of the invention and materials with catalytic functions may of course be used.\",\n \"The invention will now be explained in greater detail through the following examples.\",\n \"EXAMPLE 1 A PET nonwoven fabric (SKC50 by Hikotomi Industrial Co., Ltd., mass per unit area: 50 g/m2, pressure drop of nonwoven fabric alone: 6 Pa) was used as the base fabric, and an acrylic-based emulsion (Nipol LX820A acryl emulsion, solid portion: 22 g/m2, product of Nihon Zeon Co., Ltd.) was evenly coated onto both sides of the base fabric with a gravure coater (Asahi Precision Roll Lattice Model 35).\",\n \"A voltage of 23 kV was applied using an electrostatic tester by Kasuga Electric Works Ltd. to produce an electrostatic field.\",\n \"The electrode spacing distance was 10 cm, and the adhesive-coated base fabric (30 cm×1 m) was passed through horizontally at a speed of 1 m/min.\",\n \"GG26/60 35N (D50: 430 μm, σg: 1.2) activated carbon by Kuraray Chemical Co., Ltd. was supplied at the same time, and the activated carbon was adhered thereto.\",\n \"The adhesion of the activated carbon onto the base fabric, after removal of the moisture in the adhesive using a roller drier, was 250 g/m2 on both sides of the base fabric.\",\n \"The pressure drop of the filter element was measured to be 21 Pa by passing air through at 1 m/sec at 25° C. The pressure drop of the dried base fabric after coating of the adhesive was 7.5 Pa.\",\n \"The relationship between pressure drop and amount of the activated carbon powder adhered to the base fabric satisfied the relationship that 0≦A≦1.05 mB+C and m=0.28−0.0005D50, where 0<C≦8.The adsorptions (%) of toluene, sulfur dioxide and n-butane after 1 minute of gas flow were measured according to JIS B 9901-1997, as indices of the adsorption performance.\",\n \"The results are shown in Table 1.EXAMPLE 2 A filter element was manufactured in the same manner as Example 1, except that the activated carbon was adhered only to one side of the base fabric.\",\n \"The amount of the activated carbon powder adhered to the base fabric was 130 g/m2, the pressure drop of the filter element was 12 Pa, the pressure drop of the base fabric after coating and drying the adhesive was 7 Pa.\",\n \"The filter element satisfied the relationship that 0≦A≦1.05 mB+C and m=0.28−0.0005 D50, where 0<C≦8, was satisfied.\",\n \"The results of measurement of the adsorption performance are shown in Table 1.COMPARATIVE EXAMPLE 1 The same base fabric, adhesive and activated carbon used in Example 1 were used to manufacture a filter element by the fluidized adhesion method under the same conditions as indicated in Example 1 of Japanese Unexamined Patent Publication No.\",\n \"HEI 10-204384.As shown by the results in Table 1 including the adsorption performance, the pressure drop was large at 40 Pa. TABLE 1 Adsorption (%) Pressure Sulfur drop (Pa) Toluene dioxide n-Butane Example 1 21 95 96 91 Example 2 12 82 77 61 Comparative 40 96 96 92 Example 1 EXAMPLE 3 A filter element was manufactured in the same manner as Example 1, except that a heat-fusible fiber of nonwoven fabric (OEX by Kurashiki Textile Manufacturing Co., Ltd., fiber size: 17 dtex, mass perunit area: 64 g/m2, thickness: 1.6 mm, pressure drop of nonwoven fabric alone: 3 Pa) was used as the base fabric.\",\n \"The amount of the activated carbon adhered to the base fabric was 330 g/m2.The filter element had a pressure drop of 17.5 Pa when measured by passing 25° C. air at 1 m/sec.\",\n \"The pressure drop of the base fabric after coating and drying the adhesive was 4.5 Pa.\",\n \"The relationship between the pressure drop and the amount of the activated carbon adhered to the base fabric satisfied the relationship that 0≦A≦1.05 mB+C and m=0.28−0.0005 D50, where 0<C≦8.EXAMPLE 4 AND COMPARATIVE EXAMPLES 2-3 A nonwoven fabric (Spun Bond PK102 by Mitsui Chemical Co., Ltd., basis weight: 13 g/m2) was provided on both sides of the filter element manufactured in Example 1, and the pressure drop and adsorption were measured.\",\n \"Two automobile filters of the same type by Japan Vilene Co., Ltd. mounted in commercial automobiles were also measured for pressure drop and adsorption.\",\n \"The results are shown in Table 2.The filter of Example 4 was clearly superior in terms of pressure drop and adsorption performance.\",\n \"The filter element was also worked into a pleated shape and cut, reinforced with a comb-shaped polypropylene reinforcing material at several locations on the back of the filter element and provided with a nonwoven fabric on the front for use as an air conditioner filter.\",\n \"Prolonged stable operation was possible, and the performance as a filter was satisfactorily adequate.\",\n \"EXAMPLE 5 AND COMPARATIVE EXAMPLE 4 A nonwoven fabric (Spun Bond PK102 by Mitsui Chemical Co., Ltd., mass per unit area: 13 g/m2) was provided on both sides of the filter element manufactured in Example 1, and after further providing an electret nonwoven fabric for air filtration, the pressure drop and adsorption were measured.\",\n \"An automobile filter of the same type by Japan Vilene Co., Ltd. mounted in a commercial automobile was also measured for pressure drop and adsorption.\",\n \"The results are shown in Table 2.The filter of Example 5 was clearly superior in terms of pressure drop and adsorption performance.\",\n \"EXAMPLE 6 AND COMPARATIVE EXAMPLE 5 Activated carbon was adhered to only one side of a base fabric, and a filter element was manufactured by the same process as in Example 1.A nonwoven fabric was provided on both sides and the pressure drop and the adsorption were measured in the same manner as Example 4.For comparison, a commercial automobile-mounted automobile filter by Freudenberg Co. composed of the relatively low amount of the activated carbon adhered to the base fabric was also measured for pressure drop and adsorption (Comparative Example 5).\",\n \"The results are shown in Table 2.The filter of Example 6 was clearly superior in terms of pressure drop and adsorption performance.\",\n \"TABLE 2 Amount of the activated carbon adhered Pres- Total to the sure Adsorption (%) weight base fabric drop Sulfur n- (g/m2) (g/m2) (Pa) Toluene dioxide Butane Example 4 370 270 43.0 96.5 98.0 94.1 Comparative 373 293 82.4 92.4 95.7 73.4 Example 2 Comparative 491 417 104.0 96.4 94.5 73.3 Example 3 Example 5 390 270 78.0 96.5 98.0 94.1 Comparative 356 247 109.0 92.5 93.1 67.5 Example 4 Example 6 189 141 22.0 78.8 74.8 70.3 Comparative 512 249 43.0 86.0 86.5 54.0 Example 5 COMPARATIVE EXAMPLE 6 A filter element was manufactured in the same manner as Example 1, except that a heat-fusible fiber of nonwoven fabric (COLBACK50, CD50 by ACORDIS Co., fineness: 10-15 dtex, mass per unit area: 50 g/m2, thickness: 0.34 mm, pressure drop of nonwoven fabric alone: 6.3 Pa) was used as the base fabric.\",\n \"The amount of the activated carbon adhered to the base fabric was 210 g/m2, the pressure drop was 40 Pa, the pressure drop of the base fabric was 8 Pa after coating and drying the adhesive.\",\n \"When the thickness of the nonwoven fabric was too thin, high pressure drop resulted, relationship that 0≦A≦1.1 mB+C and m=0.28−0.0005 D50, where 0<C≦15 was not satisfied.\",\n \"EXAMPLE 7 An emboss roll set to 120° C. was used to form regular hexagonal depressions bound by a bank width of 0.2 mm, a bank height of 0.7 mm, a bank side length of 2.0 mm and a bank angle (base angle indicating base of trapezoidal cross-section) of 600 in the heat-fusible fiber of nonwoven fabric used in Example 3, on both sides of the base fabric in a turtle-shell fashion, and a filter element was manufactured in the same manner as Example 3 except for using this nonwoven fabric as the base fabric.\",\n \"The pressure drop measured in the same manner was 25 Pa, and the pressure drop of the filter manufactured in the same manner as Example 4 was measured to be 45 Pa.\",\n \"EXAMPLE 8 A filter element was manufactured by the same method using the same nonwoven fabric as in Example 3, except that activated carbon (GG26/60 35N) by Kuraray Chemical Co., Ltd. was adhered at 390 g/m2 to the top side and palladium chloride-impregnated carbon (T-E32/60) by Kuraray Chemical Co., Ltd. was adhered at 345 g/m2 to the bottom side.\",\n \"The filter element was cut to 400 mm×500 mm and housed in a 1 m3 test box.\",\n \"Air containing 50 ppm SO2 and 200 ppm CO was circulated through the filter element for 5 hours at a linear speed of 1 m/sec.\",\n \"This was repeated for 5 cycles, and the residual gas was analyzed.\",\n \"For comparison, there were prepared filter elements with activated carbon (GG26/60 35N) by Kuraray Chemical Co., Ltd. adhered at 390 g/m2 only to the top side of the filter element (Reference Example 1) and with palladium chloride-impregnated carbon (T-E32/60) by Kuraray Chemical Co., Ltd. adhered at 345 g/m2 only to the bottom side of the filter element (Reference Example 2), air containing 50 ppm SO2 and 200 ppm CO was circulated through the filter element for 5 hours at a linear speed of 1 m/sec in the same manner as before for 5 cycles, and the residual gas was analyzed.\",\n \"Absolutely no SO2 was detected in the residual gas with either of the filter elements.\",\n \"Also, compared to 100 as the CO residue rate for Reference Example 1, the CO residue rate was 80 for Reference Example 2 and 50 when activated carbon and palladium chloride-impregnated carbon were adhered, thus clearly showing an effect by the filter element with the activated carbon and palladium chloride-impregnated carbon adhered thereto.\",\n \"Industrial Applicability According to the present invention there is provided a filter element exhibiting an excellent deodorizing function and adsorbing performance with low pressure drop, as well as a method for its manufacture and a filter using the element.\",\n \"Since the filter of the invention has an excellent deodorizing function and adsorbing performance as well as low pressure drop, as mentioned above, it can be suitably used as a filter for various uses, including automobile cabin filters, indoor air purification filters and air conditioner filters, which require high deodorizing and adsorption effects with low pressure drop.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473866"}}},{"rowIdx":758,"cells":{"text":{"kind":"list like","value":[["Telephone and method for converting sound to image and displaying on screen thereof","A telephone and a method for converting sound to image and displaying on a screen thereof are disclosed.","Receiver receives a call signal.","Storage section stores call sound information.","Call sound output section outputs a call sound.","Color display device displays a color image.","Controller reads out the call sound information when the receiver receives the call signal.","Controller generates a call sound through call sound output section in response to read call sound information.","Controller analyzes read call sound information in a frame unit and acquires sound analyzed information.","Controller displays a corresponding call sound image in response to the sound analyzed information on color display device.","Accordingly, the telephone and the method for convening sound to image and displaying on a screen thereof image and display a call sound on a screen to thereby give pleasure to user and improve features of manufactured goods."],["1.A telephone comprising: a receiver for receiving a call signal; a storage section for storing call sound information; a call sound output section for outputting the call sound information; a color display device for displaying a color image; and a controller for reading the call sound information from the storage section when the receiver receives the call signal, the controller controlling an operation of the call sound output section in response to the read call sound information, the controller analyzing the read call sound information to acquire sound analyzed information, and the controller displaying a corresponding call sound image in response to the sound analyzed information on the color display device.","2.The telephone as claimed in claim 1, wherein the call sound information is synthetic sound mobile audio 2 format information, the controller acquires sound analyzed information having a program change, a channel volume, a panorama, a note, and a channel message among the synthetic sound mobile audio 2 format information, and converts the synthetic sound mobile audio 2 format information into image information having an image shape, an image size, an x axis location, a color, a y axis location, and a numerical in response to the sound analyzed information.","3.The telephone as claimed in claim 1, wherein the call sound information is voice information, the controller Fourier-transforms the voice information, acquires a pitch (fundamental frequency), a formant frequency, energy of each formant frequency from the Fourier-transformed information as sound analyzed information, and converts the Fourier-transformed information into image information having an image color, the number of images, a y axis location, and an image location in response to the sound analyzed information.","4.A telephone comprising: a receiver for receiving a call signal; a storage section for storing call sound information; a call sound output section for outputting a call sound; a gradation display device including an illuminating unit and for displaying gradation information; and.","a controller for reading the call sound information from the storage section when the receiver receives the call signal, the controller controlling an operation of the call sound output section in response to the read call sound information, the controller analyzing the read call sound information to acquire sound analyzed information, and the controller displaying a corresponding illuminating color of the illuminating unit in response to the sound analyzed information on the gradation display device.","5.The telephone as claimed in claim 4, wherein the call sound information is synthetic sound mobile audio 2 format information, the controller acquires sound analyzed information having a program change, a channel volume, and a note among the synthetic sound mobile audio 2 format information, and converts the synthetic sound mobile audio 2 format information into illumination color control information of the illuminating unit in response to the sound analyzed information.","6.The telephone as claimed in claim 4, wherein the call sound information is voice information, the controller Fourier-transforms the voice information, acquires sound analyzed information having a pitch (fundamental frequency), a formant frequency, energy of each formant frequency from the Fourier-transformed information, and converts the Fourier-transformed information into illumination color control information of the illuminating unit in response to the sound analyzed information.","7.A telephone comprising: a handset for inputting/outputting a call sound signal; a transceiver for transmitting/receiving the call sound signal; a color display device for displaying a color image; and a controller for detecting an input of the traffic signal through the handset or the transceiver, for ouputting or transmitting the input traffic signal through the handset or the transceiver, for analyzing the input traffic sound signal to acquire sound analyzed information, and for displaying a traffic sound signal corresponding to the acquired sound analyzed information on the color display.","8.The telephone as claimed in claim 7, wherein the controller Fourier-transforms the call sound information in frames, acquires a pitch (fundamental frequency), a formant frequency, energy of each formant frequency from the Fourier-transformed information as sound analyzed information, and converts the Fourier-transformed information into image information having an image color, the number of images, a y axis location, and an image size in response to the sound analyzed information 9.The telephone as claimed in claim 7, wherein the controller Fourier-transforms the call sound information in frames to obtain energy of each frame, acquires a pitch (fundamental frequency), a formant frequency, energy of each formant frequency from the Fourier-transformed information as sound analyzed information, judges a speaker's feeling index in response to the sound analyzed information, and displays character information corresponding to the judged feeling index on the color display device.","10.The telephone as claimed in claim 9, wherein the controller outputs the speaker's fundamental and reference frequencies according to the energy and pitch, outputs an average energy and a fundamentally average frequency in a predetermined time unit, and judges the sneaker's feeling change according to change rates of the average energy and the fundamentally average frequency.","11.The telephone as claimed in claim 10, wherein the average fundamental frequency is determined as an average of two fundamental frequencies except a fundamental frequency having a maximal value among the fundamental frequencies of the initial 3 frames, and when the determined average reference frequency is beyond the range from 80 Hz to 600.Hz, a greater one of the two fundamental frequency is determined as the reference frequency 12.The telephone as claimed in claim 10, wherein the predetermined time unit is three frames.","13.The telephone as claimed in claim 7, wherein the controller Fourier-transforms the call sound signal in frames to obtain energy of each frame, acquires a pitch (fundamental frequency), a formant frequency, energy of each formant frequency from the Fourier-transformed information as sound analyzed information, judges a speaker's purity degree index in response to the sound analyzed information, and displays character information corresponding to the judged purity degree index on the color display device.","14.The telephone as claimed in claim 13, wherein the controller obtains a chroma of a total color every frame and outputs a purity degree proportional to a chroma level.","15.A sound/image converting method for a call sound in a telephone, wherein the telephone includes a color display device and generates a call sound when a call signal is received or in response to a call key input, the method comprising the steps of: reading call sound data; sampling sound analyzing information of a call sound according to a format of the read call sound data; forming color image data corresponding to the sampled sound analyzing information; and synchronizing the color image data with an output of the call sound and displaying a color image on the color display device of the telephone.","16.A sound/image converting method for a call sound in a telephone, wherein the telephone includes an illuminating unit of a display device and generates a call sound when a call signal is received or in response to a call key input, the method comprising the steps of: reading call sound data; sampling sound analyzing information of a call sound according to a format of the read call sound data; forming illumination control data corresponding to the sampled sound analyzing information; and controlling an illuminating color of the illuminating unit of a display device in response to the illumination control data.","17.A sound/image converting method for a traffic sound in a telephone including a color display device, the method comprising the steps of: receiving a called party's traffic sound data; sampling sound analyzing information of a traffic sound from the called party's traffic sound data received; forming a traffic sound image corresponding to the sampled sound analyzing information; and synchronizing the traffic sound image with an output of the call sound and displaying a color image on the color display device of the telephone.","18.The method as claimed in claim 17, wherein the step of forming traffic sound image Fourier-transforming the traffic sound data in frames to obtain energy of each frame, acquiring a pitch (fundamental frequency), a formant frequency, energy of each formant frequency from the Fourier-transformed information as sound analyzed information, judging a speaker's feeling index in response to the sound analyzed information, and displaying character information corresponding to the judged feeling index on the color display device.","19.The method as claimed in claim 17, wherein the step of forming traffic sound image Fourier-transforming the traffic sound data in frames to obtain energy of each frame, acquiring a pitch (fundamental frequency), a formant frequency, energy of each formant frequency from the Fourier-transformed information as sound analyzed information, judging a speaker's purity degree index in response to the sound analyzed information, and displaying character information corresponding to the judged purity degree index on the color display device.","20.The method as claimed in claim 17, wherein the step of forming traffic sound image Fourier-transforming the traffic sound data in frames to obtain energy of each frame, obtain a pitch (fundamental frequency) from the Fourier-transformed information, displaying a color corresponding to the obtained fundamental frequency as a base color, and a waveform of a traffic sound having a circle shape on the base color through the color display device, a size of the circle shape is proportioned to a volume.","21.A telephone comprising: a handset for inputting/outputting a traffic sound signal; a transeiver for transmitting/receiving the traffic sound signal; a color display device for displaying a color image; and a controller for detecting an input of the traffic sound signal through the transceiver, outputting the input traffic sound signal to the handset, decoding the input traffic sound signal to obtain formant frequency informaiton and pitch information, and displaying a traffic sound image corresponding to the obtained formant frequency informaiton and pitch information on the color display device."],["<SOH> BACKGROUND ART <EOH>Currently, a mobile communication terminal (hereinafter, referred to as “portable phone”) such as a portable phone, a PCS phone, or an IMT-2000 phone has been spreading, and various functions of the portable phone have been introduced.","As a market competition between a manufacturer of the portable phone and a communication service company has increased, research and development through marketability and competition of an additional service has been actively undertaken.","Additional functions of the portable phone include a transmission of a character message, various bell sound services, and a game.","Suppose that a plurality of portable phones having the same call sound, that is, the same bell sound, are located in an adjacent area.","When a caller tries to call one of the portable phones, each of the portable phone users should confirm whether his/her own portable phone is being called.","In response to a portable phone user's desire to have a specific bell sound, portable phones capable of selecting various bell sounds by a user have been introduced.","Further, a portable phone receives and sets a download of a specific bell sound from a bell sound service provider or directly sets a person's voice as the bell sound.","From the beginning, a buzzer sound of a single channel was generally used for the bell sound.","Currently, an MIDI chord bell sound according to an SMAF (Synthetic music Mobile Application Format: data format for multi media contents classification provided from YAMAHA in Japan) such as 4poly YAMAHA MA1, 16poly YAMAHA MA2(MMF) has been provided.","A color screen such as a full color moving picture, graphics, an animation, etc.","is included in 2.5 generation portable phone of a CDMA 2000 1× and 3 generation portable phone of an IMT 2000.However, a portable phone user can aurally recognize a bell sound or a call sound of a portable phone but can not visibly recognize it."],["<SOH> BRIEF DESCRIPTION OF DRAWINGS <EOH>This invention will be better understood and its various objects and advantages will be more fully apply appreciated from the following description taken in conjunction with the accompanying drawings, in which: FIG.","1 is a block diagram for showing a configuration of a telephone according to an embodiment of the present invention; FIGS.","2 through 6 are tables for showing corresponding relationships of each sound analyzed information and image information with respect to a composite call sound of an MA2 format; FIG.","7 is a view for showing an image of a composite call sound in a portable phone according to an embodiment of the present invention; FIG.","8 is a flow chart for illustrating a sound/image converting method for a composite call sound in a telephone according to an embodiment of the present invention; FIG.","9 is a flow chart for illustrating a sound/image converting method for a voice call sound in a telephone according to an embodiment of the present invention; FIG.","10 is a view for showing a corresponding relationship of each sound analyzed information and image information with respect to a voice call sound and an image display thereof; FIG.","11 is an orthogonal color coordinate system for showing colors corresponding to each audio frequency when converting an audio frequency into a visible frequency according to an embodiment of the present invention; FIGS.","12 through 18 are views for showing various image display statuses of a call sound according to an embodiment of the present invention; FIG.","19 is a view for displaying the call sound by a change of a color illumination of a liquid crystal panel according to an embodiment of the present invention; FIG.","20 is a flow chart for illustrating a sound/image converting method for a traffic sound in a telephone according to an embodiment of the present invention; FIG.","21 is a view for showing an example of a corresponding relationship of emotional indexes and characters according to an embodiment of the present invention; FIG.","22 is a view for showing an example of a corresponding relationship of purity indexes and characters according to an embodiment of the present invention; FIGS.","23 and 24 are views for showing examples of displayed images with respect to the call sound according to an embodiment of the present invention.","detailed-description description=\"Detailed Description\" end=\"lead\"?"],["TECHNICAL FIELD The present invention relates to a telephone and a method for converting a sound to an image and displaying on a screen thereof, and more particularly, to a telephone and a method for converting a sound to an image and displaying on a screen thereof for displaying a bell sound of a portable phone and a call sound between a caller and a called party on a screen as a color.","BACKGROUND ART Currently, a mobile communication terminal (hereinafter, referred to as “portable phone”) such as a portable phone, a PCS phone, or an IMT-2000 phone has been spreading, and various functions of the portable phone have been introduced.","As a market competition between a manufacturer of the portable phone and a communication service company has increased, research and development through marketability and competition of an additional service has been actively undertaken.","Additional functions of the portable phone include a transmission of a character message, various bell sound services, and a game.","Suppose that a plurality of portable phones having the same call sound, that is, the same bell sound, are located in an adjacent area.","When a caller tries to call one of the portable phones, each of the portable phone users should confirm whether his/her own portable phone is being called.","In response to a portable phone user's desire to have a specific bell sound, portable phones capable of selecting various bell sounds by a user have been introduced.","Further, a portable phone receives and sets a download of a specific bell sound from a bell sound service provider or directly sets a person's voice as the bell sound.","From the beginning, a buzzer sound of a single channel was generally used for the bell sound.","Currently, an MIDI chord bell sound according to an SMAF (Synthetic music Mobile Application Format: data format for multi media contents classification provided from YAMAHA in Japan) such as 4poly YAMAHA MA1, 16poly YAMAHA MA2(MMF) has been provided.","A color screen such as a full color moving picture, graphics, an animation, etc.","is included in 2.5 generation portable phone of a CDMA 2000 1× and 3 generation portable phone of an IMT 2000.However, a portable phone user can aurally recognize a bell sound or a call sound of a portable phone but can not visibly recognize it.","DISCLOSURE OF INVENTION Accordingly, the present invention has been made to solve the above-mentioned problems occurring in the prior art.","Therefore, it is an object of the present invention to provide a telephone and a method for converting a sound to an image and displaying on a screen thereof capable of displaying a bell sound or a call sound of a telephone such as a portable phone.","It is another object of the present invention to provide a telephone and a method for converting a sound to an image and displaying on a screen thereof capable of improving the marketability of the telephone by displaying a sound as an image in the telephone.","It is another object of the present invention to provide a telephone and a method for converting a sound to an image and displaying on a screen thereof capable of displaying a called party's sound purity degree and feeling index as a character and a color image.","In order to achieve the above object, the present invention provides a telephone comprising: a receiver for receiving a call signal; a storage section for storing call sound information; a call sound output section for outputting the call sound information; a color display device for displaying a color image; and a controller for reading the call sound information from the storage section when the receiver receives the call signal, the controller controlling an operation of the call sound output section in response to the read call sound information, the controller analyzing the read call sound information to acquire sound analyzed information, and the controller displaying a corresponding call sound image in response to the sound analyzed information on the color display device.","Preferabley, the call sound information is synthetic sound mobile audio 2 format information, the controller acquires sound analyzed information having a program change, a channel volume, a panorama, a note, and a channel message among the synthetic sound mobile audio 2 format information, and converts the synthetic sound mobile audio 2 format information into image information having an image shape, an image size, an x axis location, a color, a y axis location, and a numeral in response to the sound analyzed information.","More preferably, the call sound information is voice information, the controller Fourier-transforms the voice information, acquires a pitch (fundamental frequency), a format frequency, energy of each formant frequency from the Fourier-transformed information as sound analyzed information, and converts the Fourier-transformed information into image information having an image color, the number of images, a y axis location, an image, location in response to the sound analyzed information.","Also.","there is provided a telephone comprising: a receiver for receiving a call signal; a storage section for storing call sound information; a call sound output section for outputting a call sound; a gradation display device including an illuminating unit and for displaying gradation information; and a controller for reading the call sound information from the storage section when the receiver receives the call signal, the controller controlling an operation of the call sound output section in response to the read call sound information, the controller analyzing the read call sound information to acquire sound analyzed information, and the controller displaying a corresponding illuminating color of the illuminating unit in response to the sound analyzed information on the gradation display device.","Further, there is provided a telephone comprising: a handset for inputting/outputting a call sound signal; a transceiver for transmitting/receiving the call sound signal; a color display device for displaying a color image; and a controller for detecting an input of the traffic signal through the handset or the transceiver, for ouputting or transmitting the input traffic signal through the handset or the transceiver, for analyzing the input traffic sound signal to acquire sound analyzed information, and for displaying a traffic sound signal corresponding to the acquired sound analyzed information on the color display.","Preferably, the controller Fourier-transforms the call sound information in frames, acquires a pitch (fundamental frequency), a formant frequency, energy of each formant frequency from the Fourier-transformed information as sound analyzed information, and converts the Fourier-transformed information into image information having an image color, the number of images, a y axis location, and an image location in response to the sound analyzed information.","More preferably, the controller Fourier-transforms the call sound information in frames to obtain energy of each frame, acquires a pitch (fundamental frequency), a formant frequency, energy of each formant frequency from the Fourier-transformed information as sound analyzed information, judges a speaker's feeling index in response to the sound analyzed information, and displays character information corresponding to the judged feeling index on the color display device.","Most preferably, the controller outputs the speaker's fundamental and reference frequencies according to the energy and pitch, outputs an average energy and a fundamentally average frequency in a predetermined time unit, and judges the speaker's feeling change according to change rates of the average energy and the fundamentally average frequency.","Also, the controller Fourier-transforms the call sound signal in frames to obtain energy of each frame, acquires a pitch (fundamental frequency), a formant frequency, energy of each formant frequency from the Fourier-transformed information as sound analyzed information, judges a speaker's purity degree index in response to the sound analyzed information, and displays character information corresponding to the judged purity degree index on the color display device.","Furthermore, there is provided a sound/image converting method for a call sound in a telephone, wherein the telephone includes a color display device and generates a call sound when a call signal is received or in response to a call key input, the method comprising the steps of: reading call sound data; sampling sound analyzing information of a call sound according to a format of the read call sound data; forming color image data corresponding to the sampled sound analyzing information; and synchronizing the color image data with an out put of the call sound and displaying a color image on the color display device of the telephone.","There is provided a sound/image converting method for a traffic sound in a telephone including a color display device, the method comprising the steps of: receiving a called party's traffic sound data; sampling sound analyzing information of a traffic sound from the called party's traffic sound data received; forming a traffic sound image corresponding to the sampled sound analyzing information; and synchronizing the traffic sound image with an output of the call sound and displaying a color image on the color display device of the telephone.","In an embodiment of the present invention, a traffic sound display image includes a traffic sound image, a feeling index character, and a purity degree character.","BRIEF DESCRIPTION OF DRAWINGS This invention will be better understood and its various objects and advantages will be more fully apply appreciated from the following description taken in conjunction with the accompanying drawings, in which: FIG.","1 is a block diagram for showing a configuration of a telephone according to an embodiment of the present invention; FIGS.","2 through 6 are tables for showing corresponding relationships of each sound analyzed information and image information with respect to a composite call sound of an MA2 format; FIG.","7 is a view for showing an image of a composite call sound in a portable phone according to an embodiment of the present invention; FIG.","8 is a flow chart for illustrating a sound/image converting method for a composite call sound in a telephone according to an embodiment of the present invention; FIG.","9 is a flow chart for illustrating a sound/image converting method for a voice call sound in a telephone according to an embodiment of the present invention; FIG.","10 is a view for showing a corresponding relationship of each sound analyzed information and image information with respect to a voice call sound and an image display thereof; FIG.","11 is an orthogonal color coordinate system for showing colors corresponding to each audio frequency when converting an audio frequency into a visible frequency according to an embodiment of the present invention; FIGS.","12 through 18 are views for showing various image display statuses of a call sound according to an embodiment of the present invention; FIG.","19 is a view for displaying the call sound by a change of a color illumination of a liquid crystal panel according to an embodiment of the present invention; FIG.","20 is a flow chart for illustrating a sound/image converting method for a traffic sound in a telephone according to an embodiment of the present invention; FIG.","21 is a view for showing an example of a corresponding relationship of emotional indexes and characters according to an embodiment of the present invention; FIG.","22 is a view for showing an example of a corresponding relationship of purity indexes and characters according to an embodiment of the present invention; FIGS.","23 and 24 are views for showing examples of displayed images with respect to the call sound according to an embodiment of the present invention.","BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, this invention will be described in detail with reference to the drawings.","FIG.","1 is a block diagram for showing a configuration of a portable phone according to an embodiment of the present invention.","The portable phone includes a transceiver 100, a handset 200, a display device 300, a key pad 400, a memory 500, and a controller 600.The transceiver 100 receives an RF signal transmitted from a mobile communication base station through an antenna 102 and a duplexer 104 by a receiver 106.The receiver 106 mixes the received RF signal with a frequency signal provided from a frequency synthesizer 108 and extracts a CDMA signal.","The CDMA signal is provided to a decoder 110.The decoder 110 demodulates the CDMA signal from the receiver 106 and restores the demodulated CDMA signal to channel coded data.","The decoder 110 decodes the channel coded data by a CDMA decoding algorithm and outputs packet data.","The packet data are provided to a controller 600.An encoder 112 channel-encodes the packet data provided from the controller by a CDMA encoding algorithm and provides the channel-encoded data to a transmitter 114.The transmitter 114 converts the channel-encoded data from the encoder 112 into a CDMA signal and modulates the CDMA signal.","The transmitter 114 mixes the modulated CDMA signal with the frequency signal provided from a frequency synthesizer 108 to generate an RF signal.","The RF signal is transmitted to the air through the duplexer 104 and the antenna 102.The handset 200 converts voice data and ADPCM data provided from the controller 600 into analog voice signals by a voice processor 202.A voice amplifier 204 and a speaker 206 outputs the analog voice signals from the voice processor 202 as a voice.","The voice processor 202 of the handset 200 converts a voice signal inputted through a microphone 208 into digital voice data, namely, ADPCM data and provides the converted digital voice data to the controller 600.A voice synthesizer 210 of the handset 200 synthesizes call sound data provided from the controller 200 and generates an analog call sound signal.","The analog call sound signal is outputted through the voice amplifier 204 and the speaker 206 as a bell sound.","The voice synthesizer 210 includes a voice synthesizing chip (for example, YMU759 chip of YAMAHA Co., Ltd. Japan) which processes call sound data and MIDI call sound data and generates a voice signal.","The call sound data are PCM or ADPCM for a voice and the MIDI call sound data are a computer synthesizing sound of an MA 1 or MA2 format.","A display device 300 converts image data provided from the controller 600 into LCD drive data by an LCD driver 302.The LCD drive data are provided to an LCD panel 304 and are displayed through the LCD panel as an image.","The LCD panel 304 displays the image using a gradation or color display method.","In the gradation display method, a character, a number, and an icon, etc., are displayed on a screen, and a color illumination unit 306 is used.","The color illumination unit 306 provides a color illumination to a crystal liquid panel by red and green lamps, or red, green, and blue lamps, for example, a combination of light emitting diodes.","A key pad of a general portable phone is used for a key pad 400.The key pad 400 provides a key input signal to the controller 600.The key pad 400 selects a sound/image conversion and display function, downloads call sound data, or performs a function key input for selecting an image.","A memory 500 includes a fresh memory, an random access memory (RAM), and an read only memory (ROM).","Downloaded call sound data are stored in the fresh memory.","An operating program of the portable phone is stored in the ROM.","The controller 600 includes a central processing unit 602, a vocoder 604, and a sound/image converter 606.The central processing unit 602 includes a micro computer or an interface circuit and executes the operating system of the portable phone.","The vocoder 602 receives the packet data provided from the transceiver 100.The vocoder 602 decodes compressed data by a Qualcomm code Exitation linear prediction (QCELP) decoding algorithm and provides extended sound data to the voice processor 202 of the handset 200.The vocoder 602 compresses voice data provided from the handset 200 by a QCELP encoding algorithm and generates the packet data.","The packet data is provided to the encoder 112 of the transceiver 100.In a QCELP algorithm, formant frequency information and pitch information of a voice signal are extracted, compressed, and encoded as feature information.","In a decoding operation, a voice signal similar to an original sound is generated based on received formant information and pitch information.","The sound/image converter 606 receives call and traffic sound data and generates corresponding image data by a sound/image conversion algorithm according the present invention, and provides the image data to the display device 300 through the central processing unit 602.The sound/image conversion algorithm according to the present invention will now be described as follows.","1.Image Conversion Algorithm of Composite Call Sound Having MA2 Format Sequence data of an MA2 format provided by YAMAHA Co., Ltd. are formed by a repetition of duration data of 1˜2 bytes and event data of 2 through N bytes.","The event data are divided into a control message, a note message, an NOP message, and an exclusive message according a data type.","The control message includes a channel code, a control identification (ID) a type code, and a set value, etc., in the second and third bytes.","The control message is divided into a program change, an octave shift, a channel volume, a panorama (panpot), and an expression according to the type code.","The program change is information which sets a musical instrument tone of a corresponding channel to any one among 128 kinds of musical instrument tones.","The octave shift is information which sets an octave shift amount of a corresponding channel to any one among 10 kinds of shift mounts.","The channel volume is information which sets a volume of the corresponding channel to any one grade among 32 volume grades.","The panorama (panpot) sets each decibel value of L and R stereo sound source in a stereo sound field to any one grade among 44 grades.","The expression is information which changes a volume of the corresponding channel while reproducing music.","The note message includes a channel code, an octave code, a note code, and a gate time code.","The octave code includes octave information of 4 grades having low, middle low, middle high, and high.","The note code has 12 musical scale information.","The gate time code has sound length information of a corresponding sound in a set musical scale.","The exclusive message includes a head code of 2 bytes, a message size of 1 byte, a maker ID of 1 byte, a format ID of 1 byte, data of n bytes, and a data end of 1 byte.","The exclusive message sets tone data or directly has access control information.","Accordingly, an image conversion of a MIDI call sound corresponds the above musical information to the following image information.","A Y axis of a liquid crystal display screen corresponds to a sound height, that is, a frequency of a sound.","An X axis of the liquid crystal display screen corresponds to a decibel value of a stereo sound source.","Sound analyzing information and image information are matched with each other as indicated in the following table 1.FIGS.","2 through 6 are tables for showing corresponding relationships of each piece of sound analyzed information and image information with respect to a composite call sound of an MA2 format.","With reference to FIG.","2, 4 octave ranges correspond to 4 brightness values, respectively.","With reference to FIG.","3, each musical scale in a middle high octave range has corresponding RGB value and color.","With reference to FIG.","4, panorama (panpot) values having LR stereo values correspond to coordinate values on an X axis of a screen.","With reference to FIG.","5, channel volumes correspond to image sizes.","With reference to FIG.","6, program change values correspond to image shapes.","FIG.","7 is a view for showing an image of a composite call sound in a portable phone according to an embodiment of the present invention.","With reference to FIG.","7, when 4poly synthetic call sound is formed by an ensemble sound of 4 channel musical instruments having a drum, a grand piano, a saxophone, and violin, images corresponding to each musical instrument channel are displayed by a stereo sound field in a Y axis coordinate of a screen as a sound field feeling.","Sizes of a color and an image shape with respect to sound frequency components in each channel are differently displayed according to a frequency height at the Y axis of the screen.","The image is displayed only while a sound length (duration) of each MIDI sound lasts.","With reference to FIG.","8, a central processing unit 602 checks whether a call signal is received or a traffic key is inputted (step S100).","When the call signal is not received and the traffic key is not inputted, a traffic standby status is maintained.","When checking occurs in step S100 that the call signal is received or the traffic key is inputted, the central processing unit 602 reads call sound data from the memory 500 and provides the read call sound to the voice synthesizer 210 and sound/image converter 606.That is, the central processing unit 602 reads call sound data which a user sets among various call sound data downloaded and stored in the memory (step S102).","The central processing unit 602 judges whether the read call sound data are call sound data of MA2 which a user has set among various call sound data which the user has downloaded and stored in the memory 500 (step S104).","When the read call sound data are not the call sound data of MA2, the central processing unit 602 recognizes the read sound data as voice call sound data and a voice call sound process routine is performed (step S105).","When the read call sound data are the call sound data of MA2, the image conversion algorithm of a composite call sound having an MA2 format is performed and sound analyzing information is extracted from the call sound data (step S106).","Then, an image corresponding to the extracted sound-analyzing information is formed (step S108).","TABLE 1 MA2 DATA SOUND INFORMATION IMAGE REFERENCE NOTE OCTAVE, SOUND HEIGHT Y AXIS MESSAGE NOTE VALUE LOCATION , COLOR GATE VALUE SOUND LENGTH IMAGE DISPLAY TIME CONTROL PANORAMA STEREO SOUND X AXIS MESSAGE SOURCE DECIBEL LOCATION CHANNEL VOLUME SOUND PRESSURE IMAGE DISPLAY AREA PROGRAM CHANGE INSTRUMENT NUMBER IMAGE KINF EXCLUSIVE VOICE DATA CHORD NUMBER IMAGE NUMBER 4POLY-4IMAGE MESSAGE 16POLY-16IMAGE When the formed image data is provided to the central processing unit 602 (step S110), the central processing unit 602 converts image data into RGB data.","The central processing unit 602 synthesizes the RGB data with other display data to generate image data.","The image data are provided to an LCD driver 302 and are displayed through an LCD panel 304 (step S112).","The central processing unit 602 controls the LCD panel 304 to display the image data in synchronism with an output of a call sound.","The sound/image converter 606 previously forms the tables shown in FIGS.","2 through 6 as a look-up table and generates RGB data of corresponding image according to an addressing of the sound analyzing information.","2.Image Conversion Algorithm of Voice Call Sound A person's voice or a natural sound is recorded as ADPCM or PCM data and the ADPCM or PCM data are stored in a memory.","A voice call sound information is formed as digital data sampled in a time axis so that it is outputted as a call sound when responding to a call signal.","A YMU759 chip of YAMAHA Co., Ltd. has a function which reproduces a voice call sound formed by ADPCM data and outputs a voice.","With reference to FIG.","9, when a voice call sound is received, the sound/image converter 606 receives and converts call sound information of ADPCM data into PCM data of 16 bits (step S116).","The converted PCM data is fast Fourier transformed in frames and is converted into frequency data (step S118).","Peak values and a pitch (fundamental frequency) of the frequency data are obtained every frame.","The peak values of the frequency data is filtered by a filtering algorithm.","The filtered peak values are classified in an order of energy magnitude and filtered to the predetermined number smaller than 30(step S120).","Each of the filtered peak values of the frequency data includes a formant frequency of a voice signal of a corresponding frame.","A pitch is detected from a frequency area to obtain a fundamental frequency (step S122).","The fundamental frequency is sound/image converted by a sound/image conversion algorithm to obtain a visible frequency (step S124).","The sound/image conversion algorithm in step S124 is defined by the following equation 1.Equation 1 F = F l × 2 x * ( 1 ) B F ∝ x x = log 2 ( F i F l ) + C , where, F is a visible frequency to be obtained, F1 is a reference visible frequency, an integer party of x is an octave value, BF is a brightness of a color, x is a party down a decimal point and a sound field value in one octave, fi is a sampled reference audible frequency, f1 is a reference audible frequency, and C is a constant and a real number which ranges 0≦C≦1.For example, the reference audible frequency f1 is 329.6 Hz corresponding to ‘mi’ of 4 octave.","The constant C is 0.29 by a reference frequency corresponding to ‘red’.","When a minimal audible frequency f, and a minimal visible frequency F1 are set to 20 Hz and 350 Hz, respectively, a visible frequency is 441 THz, a brightness of a color to be obtained is 4.33(43.3%) proportional to the calculated x value according the equation 1.When a color corresponding to the visible frequency obtained in step S124 and image size information corresponding to energy of each frequency are formed as image data and the formed image data are provided to the central processing unit 602 (step S126), the central processing unit 602 converts image data into RGB data.","The central processing unit 602 synthesizes the RGB data with another display data to generate image data.","The image data are provided to an LCD driver 302 and are displayed through an LCD panel 304 (step S128).","The central processing unit 602 controls the LCD panel 304 to display the image data in synchronism with an output of a voice call sound.","FIG.","10 is a view for showing a corresponding relationship of each piece of sound analyzed information and image information with respect to a voice call sound and an image display thereof.","With reference to FIG.","10, FIG.","0.10a) shows a sampled voice signal, FIG.","10b) shows a frequency spectrum of a fast Fourier transformed voice signal, and circle images corresponding to peak values of frequency data are displayed on a screen in FIG.","10c).","In FIG.","10c), each size of circles corresponding to peak values of frequency data is proportioned to the energy of each frequency.","A color of each circle is determined by the equation 1.Parts in which circles overlap with each other indicate that colors of each circle are mixed.","A color corresponding to each audible frequency has a color shown in the color coordinate system of FIG.","11 according to an octave and a musical scale.","FIGS.","12 through 18 are views for showing various image display statuses of a call sound according to an embodiment of the present invention.","In FIGS.","12 and 13, images corresponding to each sound frequency are arranged corresponding to a y axis of a screen and are displayed as areas proportional to energy amplitude of each sound frequency.","In FIG.","14, mixed colors of all frequencies are displayed from a center of a screen.","The mixed colors are displayed to be radially progressed to an edge of the screen as time goes by.","FIG.","15 shows a combination of squares having different areas according to energy magnitude of each sound frequency to be displayed as an image forming a total of a screen by a Mondrian picture form.","FIG.","16 shows sounds of 4 octaves on a sound/color conversion system circle (radial circle coordinate which corresponds a sound to a color).","The lowest octave is displayed at a square band of an outer most side.","The highest octave is displayed at a square band of an inner most side.","A musical scale of each sound is displayed as a sound corresponding to a square band.","In FIG.","17, an image such as a butterfly, a flower, and a star having a color corresponding to a musical scale of a sound to be displayed is displayed at a random coordinate.","A previously displayed image does not disappear and the next displaying image overlaps with the previously displayed image.","In FIG.","18, an image of a regular triangle having a color corresponding to a musical scale of a sound.","Furthermore, various forms such as a color bar form and a mosaic form can be used without departing from the spirit and scope of the invention.","Therefore, a portable phone user takes pleasure in a bell sound not only aurally but also visibly by a color image which rhythmically changes on a liquid crystal panel according to the bell sound.","3.Color Display of Bell Sound of Portable Phone According to Color Illuminating Unit A portable phone having a color liquid crystal panel was described in the embodiment.","However, in a portable phone having a gradation display liquid crystal panel, the bell sound can be visibly displayed.","That is, the portable phone includes a color illumination unit 306.The color illumination unit 306 includes illumination sources having at least two different colors.","The color illumination unit 306 provides a color illumination to the liquid crystal panel 304.For example, the color illumination unit 306 outputs the color illumination by a combination of a red light emitting diode, a green light emitting diode, and a blue light emitting diode.","Accordingly, in an embodiment of the present invention, in driving a color light emitting diode of the color illumination unit 306, in response to each gain information of RGB value corresponding to each sound obtained by the algorithm, a brightness of each color light emitting diode is controlled.","Thus, in response to the bell sound, as shown in FIG.","19, an illumination color is outputted to be rhythmically changed to 65000 colors on an LCD display window.","4.Image Conversion Algorithm of Traffic Sound As shown in FIG.","20, according to an image conversion display of the traffic sound, a called party's voice in a portable phone is outputted as an image and simultaneously, a feeling status and a purity according to the called party's voice analysis are expressed as characters.","The vocoder 604 decodes data and generates ADPCM data.","The ADPCM data are provided to a voice processor 202.The sound/image converter 606 samples output data of the vocoder 604 in frames to generate 1024 samples and converts them into PCM data (step S130).","An average energy Ea ( E a = 1 N ∑ i = 1 N ( E si ) 2 ) by frames is obtained (step S132).","Es is energy of each sample value, and N is the total sample number (1024) in a frame.","It is judged whether an average energy Eaj of a current frame minus an average energy Ea(j−1) of a previous frame is greater than 6 db (step S134).","When a traffic sound is present, an average energy of the initial 3 frames is obtained to calculate a reference energy Er.","Then, an average energy AEk of the next 3 frames is obtained (step S136).","Sample values of each frame are fast-Fourier-transformed (step S138).","A pitch, that is, a fundamental frequency ff is obtained based on Fourier-transformed frequency data using a known algorithm which obtains a pitch (step S140).","4-1.Voice Image Formation A fundamental frequency ff, is extracted.","A circle proportional to an amplitude of energy Eff of the extracted fundamental frequency ff is arranged at a center of a voice screen area.","A waveform at a time axis of a voice overlaps a circumference of a circle to output a voice image.","A base color of a screen is displayed as a color corresponding to the fundamental frequency ff.","The color corresponding to the fundamental frequency ff is acquired using the equation 1.As mentioned above, traffic sound image data are formed (step S142).","4-2.Feeling Index Image Formation When a person is excited, the person speaks in a loud voice according to a feeling index to thereby change a volume and the fundamental frequency.","According to an embodiment of the present invention, the feeling index is calculated based on energy values and fundamental frequencies by frames.","A reference frequency ff is determined using fundamental frequencies ff1, ff2, and ff3 of initial 3 frames.","The reference frequency fr, is determined as an average fundamental frequency ffa of the initial 3 frames.","The average fundamental frequency is determined as an average of two fundamental frequencies except a fundamental frequency having a maximal value among the fundamental frequencies of the initial 3 frames.","When the determined average reference frequency is within the range from 80 Hz to 600 Hz, the determined average reference frequency is determined as a reference frequency.","When the determined average reference frequency is beyond the range from 80 Hz to 600 Hz, the greater one of the two fundamental frequency is determined as the reference frequency fr.","When the reference energy Er and the reference frequency fr are determined, an average energy AEk and an average fundamental frequency fak are obtained in the next 3 frames.","A change rate (E rate) of an average energy obtained with respect to the reference energy Er is obtained, that is, E rate=AEk/Er.","A change rate (f rate) of an average fundamental frequency obtained with respec to the reference frequency fr is obtained, that is, f rate=fak/fr.","Feeling indexes of 10 stages are determined as shown in table 2 according to the change rate (E rate) of an average energy and the change rate (f rate) of an average fundamental frequency.","When the feeling indexes are determined, as shown in FIG.","21, character data corresponding to the feeling indexes are read to form an image (step S144).","A kind of feeling index character can be determined by a user's selection.","TABLE 2 FREQUENCY FEELING INDEX ENERGY CHANGE RATE CHANGE RATE 10 GREATER THAN 13 1.2˜3 9 11˜13 8 9˜11 7 7˜9 6 6˜7 0.8˜2 5 5˜6 4 4˜5 3 3˜4 2 0.7˜3 0.7˜1.5 1 0˜0.7 0.1˜1.2 4-3.Purity Index Image Formation In an embodiment of the present invention, the purity of a sound is displayed proportional to a chroma of a color.","That is, a clear sound is displayed close to one fundamental frequency.","An impure sound has a fundamental frequency and a harmonic frequency.","Accordingly, in a frequency spectrum per frame, the clear sound has a sampled low frequency.","The impure sound has a sampled high frequency.","The number of corresponding colors becomes larger in each frequency by the equation 1.The more a mixture thereof, the smaller a chroma of a mixed color becomes.","RGB data f1 (RGB), f2 (RGB), f3 (RGB), .",".",".","of sampled peak values per frame are obtained by the equation 1.RGB data are multiplied by energy value of each peak as a gain to obtain a sum of RGB so that a total RGB value is obtained every frame.","T(RGB)=e1f1+e2f2+e3f3+... Energy value is regulated as a value from 0 to 1.A total RGB value has chroma information of a corresponding frame indicating all colors corresponding to each frequency are mixed with each other.","Accordingly, a maximal value (MAX) and a minimal value (MIN) among each R value, each G value, and each B value are obtained from the total RGB value.","A sum (SUM) and a difference (DIF) of the maximal value (MAX) and the minimal value (MIN) are obtained.","The sum (SUM) is divided by 510 to obtain a luminance (LUM).","When the maximal value (MAX) and the minimal value (MIN) are identical with each other, a chroma is 0.When the luminance (LUM) is equal to or greater than 0.5, the chroma is determined by a value which is obtained by dividing the difference (DIF) by the sum (SUM).","When the luminance (LUM) is less than 0.5, the chroma is determined by a value which is obtained by dividing the difference (DIF) by (510−SUM).","As shown in table 3, purity indexes of 10 grades are expressed corresponding to chromas of 10 grades.","TABLE 3 CHROMA PURITY DEGREE INDEX 0˜0.1 −2 0.1˜0.2 −1 0.2˜0.3 0 0.3˜0.4 1 0.4˜0.5 2 0.5˜0.6 3 0.6˜0.7 4 0.7˜0.8 5 0.8˜0.9 6 0.9˜1.0 7 When a purity index is determined, as shown in FIG.","22, character data corresponding to the purity index are read to form purity degree image data (step S146).","A kind of a character to be displayed can be determined by a user's selection.","When a traffic sound image, a feeling index image, and a purity degree image are formed, the sound/image converter 606 provides the traffic sound image, the feeling index image, and the purity degree image to the central processing unit 602 (step S148).","The central processing unit 602 synthesizes the traffic sound image, the feeling index image, and the purity degree image from the sound/image converter 606 to form one display screen.","The one display screen is displayed on the liquid crystal panel as shown in FIGS.","20 and 21.With reference to FIG.","23, in a traffic sound display area, a called party' voice waveform having a circle shape is displayed on bright violet corresponding to a fundamental frequency (5 octave B musical scale) ground.","A character corresponding to a feeling index 7 is displayed in a feeling index display area.","A character corresponding to a purity degree 0 is displayed in a purity degree index area.","With reference to FIG.","24, in a traffic sound display area, a called party' voice waveform having a circle shape is displayed on dark violet corresponding to a fundamental frequency (3 octave B musical scale) ground.","A character corresponding to a feeling index 7 is displayed in a feeling index display area.","A character corresponding to a purity degree +2 is displayed in a purity degree index area.","A sound waveform of FIG.","24 is greater than that of FIG.","23.Accordingly, a user can visibly see the called party's volume magnitude.","Luminances of FIGS.","24 and 25 are different from each other according to a difference of the 5 octave and 3 octave.","The user can recognize whether the call party's current feeling is in an excited or quiet state.","The user can recognize whether the called party's voice is clear or impure through a character and an index.","5.Another Example of an Image Conversion of a Traffic Sound Formant frequency information and pitch frequency information extracted in a decoding operation of the vocoder 104 are provided to a sound/image converter 606.A color corresponding to the formant frequency information and pitch frequency information is obtained by the equation 1.In the case, it is unnecessary for the sound/image converter 606 to perform a fast-Fourier transformation operation.","Further, a sound/image converter transplants a program in a controller and forms it with a software.","The sound/image converter can be integrated to a custom semiconductor.","A sound/image converter and a voice synthesizer can be hardware embodied by a custom semiconductor.","As mentioned above, according to the present invention, since a call sound of a portable phone, that is, a bell sound is audibly outputted and is visibly displayed on a screen, a newly added function is provided to the portable phone to improve a marketability.","Also, since a called party's voice is displayed as an image, the called party's feeling and voice purity degree are displayed, additional pleasure is provided to the portable phone user, and marketability of the product is improved.","While this invention has been particularly shown and described with reference to particular embodiments thereof, it will be understood by those skilled in the art that various changes in form and details may be effected therein without departing from the spirit and scope of the invention as defined by the appended claims."]],"string":"[\n [\n \"Telephone and method for converting sound to image and displaying on screen thereof\",\n \"A telephone and a method for converting sound to image and displaying on a screen thereof are disclosed.\",\n \"Receiver receives a call signal.\",\n \"Storage section stores call sound information.\",\n \"Call sound output section outputs a call sound.\",\n \"Color display device displays a color image.\",\n \"Controller reads out the call sound information when the receiver receives the call signal.\",\n \"Controller generates a call sound through call sound output section in response to read call sound information.\",\n \"Controller analyzes read call sound information in a frame unit and acquires sound analyzed information.\",\n \"Controller displays a corresponding call sound image in response to the sound analyzed information on color display device.\",\n \"Accordingly, the telephone and the method for convening sound to image and displaying on a screen thereof image and display a call sound on a screen to thereby give pleasure to user and improve features of manufactured goods.\"\n ],\n [\n \"1.A telephone comprising: a receiver for receiving a call signal; a storage section for storing call sound information; a call sound output section for outputting the call sound information; a color display device for displaying a color image; and a controller for reading the call sound information from the storage section when the receiver receives the call signal, the controller controlling an operation of the call sound output section in response to the read call sound information, the controller analyzing the read call sound information to acquire sound analyzed information, and the controller displaying a corresponding call sound image in response to the sound analyzed information on the color display device.\",\n \"2.The telephone as claimed in claim 1, wherein the call sound information is synthetic sound mobile audio 2 format information, the controller acquires sound analyzed information having a program change, a channel volume, a panorama, a note, and a channel message among the synthetic sound mobile audio 2 format information, and converts the synthetic sound mobile audio 2 format information into image information having an image shape, an image size, an x axis location, a color, a y axis location, and a numerical in response to the sound analyzed information.\",\n \"3.The telephone as claimed in claim 1, wherein the call sound information is voice information, the controller Fourier-transforms the voice information, acquires a pitch (fundamental frequency), a formant frequency, energy of each formant frequency from the Fourier-transformed information as sound analyzed information, and converts the Fourier-transformed information into image information having an image color, the number of images, a y axis location, and an image location in response to the sound analyzed information.\",\n \"4.A telephone comprising: a receiver for receiving a call signal; a storage section for storing call sound information; a call sound output section for outputting a call sound; a gradation display device including an illuminating unit and for displaying gradation information; and.\",\n \"a controller for reading the call sound information from the storage section when the receiver receives the call signal, the controller controlling an operation of the call sound output section in response to the read call sound information, the controller analyzing the read call sound information to acquire sound analyzed information, and the controller displaying a corresponding illuminating color of the illuminating unit in response to the sound analyzed information on the gradation display device.\",\n \"5.The telephone as claimed in claim 4, wherein the call sound information is synthetic sound mobile audio 2 format information, the controller acquires sound analyzed information having a program change, a channel volume, and a note among the synthetic sound mobile audio 2 format information, and converts the synthetic sound mobile audio 2 format information into illumination color control information of the illuminating unit in response to the sound analyzed information.\",\n \"6.The telephone as claimed in claim 4, wherein the call sound information is voice information, the controller Fourier-transforms the voice information, acquires sound analyzed information having a pitch (fundamental frequency), a formant frequency, energy of each formant frequency from the Fourier-transformed information, and converts the Fourier-transformed information into illumination color control information of the illuminating unit in response to the sound analyzed information.\",\n \"7.A telephone comprising: a handset for inputting/outputting a call sound signal; a transceiver for transmitting/receiving the call sound signal; a color display device for displaying a color image; and a controller for detecting an input of the traffic signal through the handset or the transceiver, for ouputting or transmitting the input traffic signal through the handset or the transceiver, for analyzing the input traffic sound signal to acquire sound analyzed information, and for displaying a traffic sound signal corresponding to the acquired sound analyzed information on the color display.\",\n \"8.The telephone as claimed in claim 7, wherein the controller Fourier-transforms the call sound information in frames, acquires a pitch (fundamental frequency), a formant frequency, energy of each formant frequency from the Fourier-transformed information as sound analyzed information, and converts the Fourier-transformed information into image information having an image color, the number of images, a y axis location, and an image size in response to the sound analyzed information 9.The telephone as claimed in claim 7, wherein the controller Fourier-transforms the call sound information in frames to obtain energy of each frame, acquires a pitch (fundamental frequency), a formant frequency, energy of each formant frequency from the Fourier-transformed information as sound analyzed information, judges a speaker's feeling index in response to the sound analyzed information, and displays character information corresponding to the judged feeling index on the color display device.\",\n \"10.The telephone as claimed in claim 9, wherein the controller outputs the speaker's fundamental and reference frequencies according to the energy and pitch, outputs an average energy and a fundamentally average frequency in a predetermined time unit, and judges the sneaker's feeling change according to change rates of the average energy and the fundamentally average frequency.\",\n \"11.The telephone as claimed in claim 10, wherein the average fundamental frequency is determined as an average of two fundamental frequencies except a fundamental frequency having a maximal value among the fundamental frequencies of the initial 3 frames, and when the determined average reference frequency is beyond the range from 80 Hz to 600.Hz, a greater one of the two fundamental frequency is determined as the reference frequency 12.The telephone as claimed in claim 10, wherein the predetermined time unit is three frames.\",\n \"13.The telephone as claimed in claim 7, wherein the controller Fourier-transforms the call sound signal in frames to obtain energy of each frame, acquires a pitch (fundamental frequency), a formant frequency, energy of each formant frequency from the Fourier-transformed information as sound analyzed information, judges a speaker's purity degree index in response to the sound analyzed information, and displays character information corresponding to the judged purity degree index on the color display device.\",\n \"14.The telephone as claimed in claim 13, wherein the controller obtains a chroma of a total color every frame and outputs a purity degree proportional to a chroma level.\",\n \"15.A sound/image converting method for a call sound in a telephone, wherein the telephone includes a color display device and generates a call sound when a call signal is received or in response to a call key input, the method comprising the steps of: reading call sound data; sampling sound analyzing information of a call sound according to a format of the read call sound data; forming color image data corresponding to the sampled sound analyzing information; and synchronizing the color image data with an output of the call sound and displaying a color image on the color display device of the telephone.\",\n \"16.A sound/image converting method for a call sound in a telephone, wherein the telephone includes an illuminating unit of a display device and generates a call sound when a call signal is received or in response to a call key input, the method comprising the steps of: reading call sound data; sampling sound analyzing information of a call sound according to a format of the read call sound data; forming illumination control data corresponding to the sampled sound analyzing information; and controlling an illuminating color of the illuminating unit of a display device in response to the illumination control data.\",\n \"17.A sound/image converting method for a traffic sound in a telephone including a color display device, the method comprising the steps of: receiving a called party's traffic sound data; sampling sound analyzing information of a traffic sound from the called party's traffic sound data received; forming a traffic sound image corresponding to the sampled sound analyzing information; and synchronizing the traffic sound image with an output of the call sound and displaying a color image on the color display device of the telephone.\",\n \"18.The method as claimed in claim 17, wherein the step of forming traffic sound image Fourier-transforming the traffic sound data in frames to obtain energy of each frame, acquiring a pitch (fundamental frequency), a formant frequency, energy of each formant frequency from the Fourier-transformed information as sound analyzed information, judging a speaker's feeling index in response to the sound analyzed information, and displaying character information corresponding to the judged feeling index on the color display device.\",\n \"19.The method as claimed in claim 17, wherein the step of forming traffic sound image Fourier-transforming the traffic sound data in frames to obtain energy of each frame, acquiring a pitch (fundamental frequency), a formant frequency, energy of each formant frequency from the Fourier-transformed information as sound analyzed information, judging a speaker's purity degree index in response to the sound analyzed information, and displaying character information corresponding to the judged purity degree index on the color display device.\",\n \"20.The method as claimed in claim 17, wherein the step of forming traffic sound image Fourier-transforming the traffic sound data in frames to obtain energy of each frame, obtain a pitch (fundamental frequency) from the Fourier-transformed information, displaying a color corresponding to the obtained fundamental frequency as a base color, and a waveform of a traffic sound having a circle shape on the base color through the color display device, a size of the circle shape is proportioned to a volume.\",\n \"21.A telephone comprising: a handset for inputting/outputting a traffic sound signal; a transeiver for transmitting/receiving the traffic sound signal; a color display device for displaying a color image; and a controller for detecting an input of the traffic sound signal through the transceiver, outputting the input traffic sound signal to the handset, decoding the input traffic sound signal to obtain formant frequency informaiton and pitch information, and displaying a traffic sound image corresponding to the obtained formant frequency informaiton and pitch information on the color display device.\"\n ],\n [\n \"<SOH> BACKGROUND ART <EOH>Currently, a mobile communication terminal (hereinafter, referred to as “portable phone”) such as a portable phone, a PCS phone, or an IMT-2000 phone has been spreading, and various functions of the portable phone have been introduced.\",\n \"As a market competition between a manufacturer of the portable phone and a communication service company has increased, research and development through marketability and competition of an additional service has been actively undertaken.\",\n \"Additional functions of the portable phone include a transmission of a character message, various bell sound services, and a game.\",\n \"Suppose that a plurality of portable phones having the same call sound, that is, the same bell sound, are located in an adjacent area.\",\n \"When a caller tries to call one of the portable phones, each of the portable phone users should confirm whether his/her own portable phone is being called.\",\n \"In response to a portable phone user's desire to have a specific bell sound, portable phones capable of selecting various bell sounds by a user have been introduced.\",\n \"Further, a portable phone receives and sets a download of a specific bell sound from a bell sound service provider or directly sets a person's voice as the bell sound.\",\n \"From the beginning, a buzzer sound of a single channel was generally used for the bell sound.\",\n \"Currently, an MIDI chord bell sound according to an SMAF (Synthetic music Mobile Application Format: data format for multi media contents classification provided from YAMAHA in Japan) such as 4poly YAMAHA MA1, 16poly YAMAHA MA2(MMF) has been provided.\",\n \"A color screen such as a full color moving picture, graphics, an animation, etc.\",\n \"is included in 2.5 generation portable phone of a CDMA 2000 1× and 3 generation portable phone of an IMT 2000.However, a portable phone user can aurally recognize a bell sound or a call sound of a portable phone but can not visibly recognize it.\"\n ],\n [\n \"<SOH> BRIEF DESCRIPTION OF DRAWINGS <EOH>This invention will be better understood and its various objects and advantages will be more fully apply appreciated from the following description taken in conjunction with the accompanying drawings, in which: FIG.\",\n \"1 is a block diagram for showing a configuration of a telephone according to an embodiment of the present invention; FIGS.\",\n \"2 through 6 are tables for showing corresponding relationships of each sound analyzed information and image information with respect to a composite call sound of an MA2 format; FIG.\",\n \"7 is a view for showing an image of a composite call sound in a portable phone according to an embodiment of the present invention; FIG.\",\n \"8 is a flow chart for illustrating a sound/image converting method for a composite call sound in a telephone according to an embodiment of the present invention; FIG.\",\n \"9 is a flow chart for illustrating a sound/image converting method for a voice call sound in a telephone according to an embodiment of the present invention; FIG.\",\n \"10 is a view for showing a corresponding relationship of each sound analyzed information and image information with respect to a voice call sound and an image display thereof; FIG.\",\n \"11 is an orthogonal color coordinate system for showing colors corresponding to each audio frequency when converting an audio frequency into a visible frequency according to an embodiment of the present invention; FIGS.\",\n \"12 through 18 are views for showing various image display statuses of a call sound according to an embodiment of the present invention; FIG.\",\n \"19 is a view for displaying the call sound by a change of a color illumination of a liquid crystal panel according to an embodiment of the present invention; FIG.\",\n \"20 is a flow chart for illustrating a sound/image converting method for a traffic sound in a telephone according to an embodiment of the present invention; FIG.\",\n \"21 is a view for showing an example of a corresponding relationship of emotional indexes and characters according to an embodiment of the present invention; FIG.\",\n \"22 is a view for showing an example of a corresponding relationship of purity indexes and characters according to an embodiment of the present invention; FIGS.\",\n \"23 and 24 are views for showing examples of displayed images with respect to the call sound according to an embodiment of the present invention.\",\n \"detailed-description description=\\\"Detailed Description\\\" end=\\\"lead\\\"?\"\n ],\n [\n \"TECHNICAL FIELD The present invention relates to a telephone and a method for converting a sound to an image and displaying on a screen thereof, and more particularly, to a telephone and a method for converting a sound to an image and displaying on a screen thereof for displaying a bell sound of a portable phone and a call sound between a caller and a called party on a screen as a color.\",\n \"BACKGROUND ART Currently, a mobile communication terminal (hereinafter, referred to as “portable phone”) such as a portable phone, a PCS phone, or an IMT-2000 phone has been spreading, and various functions of the portable phone have been introduced.\",\n \"As a market competition between a manufacturer of the portable phone and a communication service company has increased, research and development through marketability and competition of an additional service has been actively undertaken.\",\n \"Additional functions of the portable phone include a transmission of a character message, various bell sound services, and a game.\",\n \"Suppose that a plurality of portable phones having the same call sound, that is, the same bell sound, are located in an adjacent area.\",\n \"When a caller tries to call one of the portable phones, each of the portable phone users should confirm whether his/her own portable phone is being called.\",\n \"In response to a portable phone user's desire to have a specific bell sound, portable phones capable of selecting various bell sounds by a user have been introduced.\",\n \"Further, a portable phone receives and sets a download of a specific bell sound from a bell sound service provider or directly sets a person's voice as the bell sound.\",\n \"From the beginning, a buzzer sound of a single channel was generally used for the bell sound.\",\n \"Currently, an MIDI chord bell sound according to an SMAF (Synthetic music Mobile Application Format: data format for multi media contents classification provided from YAMAHA in Japan) such as 4poly YAMAHA MA1, 16poly YAMAHA MA2(MMF) has been provided.\",\n \"A color screen such as a full color moving picture, graphics, an animation, etc.\",\n \"is included in 2.5 generation portable phone of a CDMA 2000 1× and 3 generation portable phone of an IMT 2000.However, a portable phone user can aurally recognize a bell sound or a call sound of a portable phone but can not visibly recognize it.\",\n \"DISCLOSURE OF INVENTION Accordingly, the present invention has been made to solve the above-mentioned problems occurring in the prior art.\",\n \"Therefore, it is an object of the present invention to provide a telephone and a method for converting a sound to an image and displaying on a screen thereof capable of displaying a bell sound or a call sound of a telephone such as a portable phone.\",\n \"It is another object of the present invention to provide a telephone and a method for converting a sound to an image and displaying on a screen thereof capable of improving the marketability of the telephone by displaying a sound as an image in the telephone.\",\n \"It is another object of the present invention to provide a telephone and a method for converting a sound to an image and displaying on a screen thereof capable of displaying a called party's sound purity degree and feeling index as a character and a color image.\",\n \"In order to achieve the above object, the present invention provides a telephone comprising: a receiver for receiving a call signal; a storage section for storing call sound information; a call sound output section for outputting the call sound information; a color display device for displaying a color image; and a controller for reading the call sound information from the storage section when the receiver receives the call signal, the controller controlling an operation of the call sound output section in response to the read call sound information, the controller analyzing the read call sound information to acquire sound analyzed information, and the controller displaying a corresponding call sound image in response to the sound analyzed information on the color display device.\",\n \"Preferabley, the call sound information is synthetic sound mobile audio 2 format information, the controller acquires sound analyzed information having a program change, a channel volume, a panorama, a note, and a channel message among the synthetic sound mobile audio 2 format information, and converts the synthetic sound mobile audio 2 format information into image information having an image shape, an image size, an x axis location, a color, a y axis location, and a numeral in response to the sound analyzed information.\",\n \"More preferably, the call sound information is voice information, the controller Fourier-transforms the voice information, acquires a pitch (fundamental frequency), a format frequency, energy of each formant frequency from the Fourier-transformed information as sound analyzed information, and converts the Fourier-transformed information into image information having an image color, the number of images, a y axis location, an image, location in response to the sound analyzed information.\",\n \"Also.\",\n \"there is provided a telephone comprising: a receiver for receiving a call signal; a storage section for storing call sound information; a call sound output section for outputting a call sound; a gradation display device including an illuminating unit and for displaying gradation information; and a controller for reading the call sound information from the storage section when the receiver receives the call signal, the controller controlling an operation of the call sound output section in response to the read call sound information, the controller analyzing the read call sound information to acquire sound analyzed information, and the controller displaying a corresponding illuminating color of the illuminating unit in response to the sound analyzed information on the gradation display device.\",\n \"Further, there is provided a telephone comprising: a handset for inputting/outputting a call sound signal; a transceiver for transmitting/receiving the call sound signal; a color display device for displaying a color image; and a controller for detecting an input of the traffic signal through the handset or the transceiver, for ouputting or transmitting the input traffic signal through the handset or the transceiver, for analyzing the input traffic sound signal to acquire sound analyzed information, and for displaying a traffic sound signal corresponding to the acquired sound analyzed information on the color display.\",\n \"Preferably, the controller Fourier-transforms the call sound information in frames, acquires a pitch (fundamental frequency), a formant frequency, energy of each formant frequency from the Fourier-transformed information as sound analyzed information, and converts the Fourier-transformed information into image information having an image color, the number of images, a y axis location, and an image location in response to the sound analyzed information.\",\n \"More preferably, the controller Fourier-transforms the call sound information in frames to obtain energy of each frame, acquires a pitch (fundamental frequency), a formant frequency, energy of each formant frequency from the Fourier-transformed information as sound analyzed information, judges a speaker's feeling index in response to the sound analyzed information, and displays character information corresponding to the judged feeling index on the color display device.\",\n \"Most preferably, the controller outputs the speaker's fundamental and reference frequencies according to the energy and pitch, outputs an average energy and a fundamentally average frequency in a predetermined time unit, and judges the speaker's feeling change according to change rates of the average energy and the fundamentally average frequency.\",\n \"Also, the controller Fourier-transforms the call sound signal in frames to obtain energy of each frame, acquires a pitch (fundamental frequency), a formant frequency, energy of each formant frequency from the Fourier-transformed information as sound analyzed information, judges a speaker's purity degree index in response to the sound analyzed information, and displays character information corresponding to the judged purity degree index on the color display device.\",\n \"Furthermore, there is provided a sound/image converting method for a call sound in a telephone, wherein the telephone includes a color display device and generates a call sound when a call signal is received or in response to a call key input, the method comprising the steps of: reading call sound data; sampling sound analyzing information of a call sound according to a format of the read call sound data; forming color image data corresponding to the sampled sound analyzing information; and synchronizing the color image data with an out put of the call sound and displaying a color image on the color display device of the telephone.\",\n \"There is provided a sound/image converting method for a traffic sound in a telephone including a color display device, the method comprising the steps of: receiving a called party's traffic sound data; sampling sound analyzing information of a traffic sound from the called party's traffic sound data received; forming a traffic sound image corresponding to the sampled sound analyzing information; and synchronizing the traffic sound image with an output of the call sound and displaying a color image on the color display device of the telephone.\",\n \"In an embodiment of the present invention, a traffic sound display image includes a traffic sound image, a feeling index character, and a purity degree character.\",\n \"BRIEF DESCRIPTION OF DRAWINGS This invention will be better understood and its various objects and advantages will be more fully apply appreciated from the following description taken in conjunction with the accompanying drawings, in which: FIG.\",\n \"1 is a block diagram for showing a configuration of a telephone according to an embodiment of the present invention; FIGS.\",\n \"2 through 6 are tables for showing corresponding relationships of each sound analyzed information and image information with respect to a composite call sound of an MA2 format; FIG.\",\n \"7 is a view for showing an image of a composite call sound in a portable phone according to an embodiment of the present invention; FIG.\",\n \"8 is a flow chart for illustrating a sound/image converting method for a composite call sound in a telephone according to an embodiment of the present invention; FIG.\",\n \"9 is a flow chart for illustrating a sound/image converting method for a voice call sound in a telephone according to an embodiment of the present invention; FIG.\",\n \"10 is a view for showing a corresponding relationship of each sound analyzed information and image information with respect to a voice call sound and an image display thereof; FIG.\",\n \"11 is an orthogonal color coordinate system for showing colors corresponding to each audio frequency when converting an audio frequency into a visible frequency according to an embodiment of the present invention; FIGS.\",\n \"12 through 18 are views for showing various image display statuses of a call sound according to an embodiment of the present invention; FIG.\",\n \"19 is a view for displaying the call sound by a change of a color illumination of a liquid crystal panel according to an embodiment of the present invention; FIG.\",\n \"20 is a flow chart for illustrating a sound/image converting method for a traffic sound in a telephone according to an embodiment of the present invention; FIG.\",\n \"21 is a view for showing an example of a corresponding relationship of emotional indexes and characters according to an embodiment of the present invention; FIG.\",\n \"22 is a view for showing an example of a corresponding relationship of purity indexes and characters according to an embodiment of the present invention; FIGS.\",\n \"23 and 24 are views for showing examples of displayed images with respect to the call sound according to an embodiment of the present invention.\",\n \"BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, this invention will be described in detail with reference to the drawings.\",\n \"FIG.\",\n \"1 is a block diagram for showing a configuration of a portable phone according to an embodiment of the present invention.\",\n \"The portable phone includes a transceiver 100, a handset 200, a display device 300, a key pad 400, a memory 500, and a controller 600.The transceiver 100 receives an RF signal transmitted from a mobile communication base station through an antenna 102 and a duplexer 104 by a receiver 106.The receiver 106 mixes the received RF signal with a frequency signal provided from a frequency synthesizer 108 and extracts a CDMA signal.\",\n \"The CDMA signal is provided to a decoder 110.The decoder 110 demodulates the CDMA signal from the receiver 106 and restores the demodulated CDMA signal to channel coded data.\",\n \"The decoder 110 decodes the channel coded data by a CDMA decoding algorithm and outputs packet data.\",\n \"The packet data are provided to a controller 600.An encoder 112 channel-encodes the packet data provided from the controller by a CDMA encoding algorithm and provides the channel-encoded data to a transmitter 114.The transmitter 114 converts the channel-encoded data from the encoder 112 into a CDMA signal and modulates the CDMA signal.\",\n \"The transmitter 114 mixes the modulated CDMA signal with the frequency signal provided from a frequency synthesizer 108 to generate an RF signal.\",\n \"The RF signal is transmitted to the air through the duplexer 104 and the antenna 102.The handset 200 converts voice data and ADPCM data provided from the controller 600 into analog voice signals by a voice processor 202.A voice amplifier 204 and a speaker 206 outputs the analog voice signals from the voice processor 202 as a voice.\",\n \"The voice processor 202 of the handset 200 converts a voice signal inputted through a microphone 208 into digital voice data, namely, ADPCM data and provides the converted digital voice data to the controller 600.A voice synthesizer 210 of the handset 200 synthesizes call sound data provided from the controller 200 and generates an analog call sound signal.\",\n \"The analog call sound signal is outputted through the voice amplifier 204 and the speaker 206 as a bell sound.\",\n \"The voice synthesizer 210 includes a voice synthesizing chip (for example, YMU759 chip of YAMAHA Co., Ltd. Japan) which processes call sound data and MIDI call sound data and generates a voice signal.\",\n \"The call sound data are PCM or ADPCM for a voice and the MIDI call sound data are a computer synthesizing sound of an MA 1 or MA2 format.\",\n \"A display device 300 converts image data provided from the controller 600 into LCD drive data by an LCD driver 302.The LCD drive data are provided to an LCD panel 304 and are displayed through the LCD panel as an image.\",\n \"The LCD panel 304 displays the image using a gradation or color display method.\",\n \"In the gradation display method, a character, a number, and an icon, etc., are displayed on a screen, and a color illumination unit 306 is used.\",\n \"The color illumination unit 306 provides a color illumination to a crystal liquid panel by red and green lamps, or red, green, and blue lamps, for example, a combination of light emitting diodes.\",\n \"A key pad of a general portable phone is used for a key pad 400.The key pad 400 provides a key input signal to the controller 600.The key pad 400 selects a sound/image conversion and display function, downloads call sound data, or performs a function key input for selecting an image.\",\n \"A memory 500 includes a fresh memory, an random access memory (RAM), and an read only memory (ROM).\",\n \"Downloaded call sound data are stored in the fresh memory.\",\n \"An operating program of the portable phone is stored in the ROM.\",\n \"The controller 600 includes a central processing unit 602, a vocoder 604, and a sound/image converter 606.The central processing unit 602 includes a micro computer or an interface circuit and executes the operating system of the portable phone.\",\n \"The vocoder 602 receives the packet data provided from the transceiver 100.The vocoder 602 decodes compressed data by a Qualcomm code Exitation linear prediction (QCELP) decoding algorithm and provides extended sound data to the voice processor 202 of the handset 200.The vocoder 602 compresses voice data provided from the handset 200 by a QCELP encoding algorithm and generates the packet data.\",\n \"The packet data is provided to the encoder 112 of the transceiver 100.In a QCELP algorithm, formant frequency information and pitch information of a voice signal are extracted, compressed, and encoded as feature information.\",\n \"In a decoding operation, a voice signal similar to an original sound is generated based on received formant information and pitch information.\",\n \"The sound/image converter 606 receives call and traffic sound data and generates corresponding image data by a sound/image conversion algorithm according the present invention, and provides the image data to the display device 300 through the central processing unit 602.The sound/image conversion algorithm according to the present invention will now be described as follows.\",\n \"1.Image Conversion Algorithm of Composite Call Sound Having MA2 Format Sequence data of an MA2 format provided by YAMAHA Co., Ltd. are formed by a repetition of duration data of 1˜2 bytes and event data of 2 through N bytes.\",\n \"The event data are divided into a control message, a note message, an NOP message, and an exclusive message according a data type.\",\n \"The control message includes a channel code, a control identification (ID) a type code, and a set value, etc., in the second and third bytes.\",\n \"The control message is divided into a program change, an octave shift, a channel volume, a panorama (panpot), and an expression according to the type code.\",\n \"The program change is information which sets a musical instrument tone of a corresponding channel to any one among 128 kinds of musical instrument tones.\",\n \"The octave shift is information which sets an octave shift amount of a corresponding channel to any one among 10 kinds of shift mounts.\",\n \"The channel volume is information which sets a volume of the corresponding channel to any one grade among 32 volume grades.\",\n \"The panorama (panpot) sets each decibel value of L and R stereo sound source in a stereo sound field to any one grade among 44 grades.\",\n \"The expression is information which changes a volume of the corresponding channel while reproducing music.\",\n \"The note message includes a channel code, an octave code, a note code, and a gate time code.\",\n \"The octave code includes octave information of 4 grades having low, middle low, middle high, and high.\",\n \"The note code has 12 musical scale information.\",\n \"The gate time code has sound length information of a corresponding sound in a set musical scale.\",\n \"The exclusive message includes a head code of 2 bytes, a message size of 1 byte, a maker ID of 1 byte, a format ID of 1 byte, data of n bytes, and a data end of 1 byte.\",\n \"The exclusive message sets tone data or directly has access control information.\",\n \"Accordingly, an image conversion of a MIDI call sound corresponds the above musical information to the following image information.\",\n \"A Y axis of a liquid crystal display screen corresponds to a sound height, that is, a frequency of a sound.\",\n \"An X axis of the liquid crystal display screen corresponds to a decibel value of a stereo sound source.\",\n \"Sound analyzing information and image information are matched with each other as indicated in the following table 1.FIGS.\",\n \"2 through 6 are tables for showing corresponding relationships of each piece of sound analyzed information and image information with respect to a composite call sound of an MA2 format.\",\n \"With reference to FIG.\",\n \"2, 4 octave ranges correspond to 4 brightness values, respectively.\",\n \"With reference to FIG.\",\n \"3, each musical scale in a middle high octave range has corresponding RGB value and color.\",\n \"With reference to FIG.\",\n \"4, panorama (panpot) values having LR stereo values correspond to coordinate values on an X axis of a screen.\",\n \"With reference to FIG.\",\n \"5, channel volumes correspond to image sizes.\",\n \"With reference to FIG.\",\n \"6, program change values correspond to image shapes.\",\n \"FIG.\",\n \"7 is a view for showing an image of a composite call sound in a portable phone according to an embodiment of the present invention.\",\n \"With reference to FIG.\",\n \"7, when 4poly synthetic call sound is formed by an ensemble sound of 4 channel musical instruments having a drum, a grand piano, a saxophone, and violin, images corresponding to each musical instrument channel are displayed by a stereo sound field in a Y axis coordinate of a screen as a sound field feeling.\",\n \"Sizes of a color and an image shape with respect to sound frequency components in each channel are differently displayed according to a frequency height at the Y axis of the screen.\",\n \"The image is displayed only while a sound length (duration) of each MIDI sound lasts.\",\n \"With reference to FIG.\",\n \"8, a central processing unit 602 checks whether a call signal is received or a traffic key is inputted (step S100).\",\n \"When the call signal is not received and the traffic key is not inputted, a traffic standby status is maintained.\",\n \"When checking occurs in step S100 that the call signal is received or the traffic key is inputted, the central processing unit 602 reads call sound data from the memory 500 and provides the read call sound to the voice synthesizer 210 and sound/image converter 606.That is, the central processing unit 602 reads call sound data which a user sets among various call sound data downloaded and stored in the memory (step S102).\",\n \"The central processing unit 602 judges whether the read call sound data are call sound data of MA2 which a user has set among various call sound data which the user has downloaded and stored in the memory 500 (step S104).\",\n \"When the read call sound data are not the call sound data of MA2, the central processing unit 602 recognizes the read sound data as voice call sound data and a voice call sound process routine is performed (step S105).\",\n \"When the read call sound data are the call sound data of MA2, the image conversion algorithm of a composite call sound having an MA2 format is performed and sound analyzing information is extracted from the call sound data (step S106).\",\n \"Then, an image corresponding to the extracted sound-analyzing information is formed (step S108).\",\n \"TABLE 1 MA2 DATA SOUND INFORMATION IMAGE REFERENCE NOTE OCTAVE, SOUND HEIGHT Y AXIS MESSAGE NOTE VALUE LOCATION , COLOR GATE VALUE SOUND LENGTH IMAGE DISPLAY TIME CONTROL PANORAMA STEREO SOUND X AXIS MESSAGE SOURCE DECIBEL LOCATION CHANNEL VOLUME SOUND PRESSURE IMAGE DISPLAY AREA PROGRAM CHANGE INSTRUMENT NUMBER IMAGE KINF EXCLUSIVE VOICE DATA CHORD NUMBER IMAGE NUMBER 4POLY-4IMAGE MESSAGE 16POLY-16IMAGE When the formed image data is provided to the central processing unit 602 (step S110), the central processing unit 602 converts image data into RGB data.\",\n \"The central processing unit 602 synthesizes the RGB data with other display data to generate image data.\",\n \"The image data are provided to an LCD driver 302 and are displayed through an LCD panel 304 (step S112).\",\n \"The central processing unit 602 controls the LCD panel 304 to display the image data in synchronism with an output of a call sound.\",\n \"The sound/image converter 606 previously forms the tables shown in FIGS.\",\n \"2 through 6 as a look-up table and generates RGB data of corresponding image according to an addressing of the sound analyzing information.\",\n \"2.Image Conversion Algorithm of Voice Call Sound A person's voice or a natural sound is recorded as ADPCM or PCM data and the ADPCM or PCM data are stored in a memory.\",\n \"A voice call sound information is formed as digital data sampled in a time axis so that it is outputted as a call sound when responding to a call signal.\",\n \"A YMU759 chip of YAMAHA Co., Ltd. has a function which reproduces a voice call sound formed by ADPCM data and outputs a voice.\",\n \"With reference to FIG.\",\n \"9, when a voice call sound is received, the sound/image converter 606 receives and converts call sound information of ADPCM data into PCM data of 16 bits (step S116).\",\n \"The converted PCM data is fast Fourier transformed in frames and is converted into frequency data (step S118).\",\n \"Peak values and a pitch (fundamental frequency) of the frequency data are obtained every frame.\",\n \"The peak values of the frequency data is filtered by a filtering algorithm.\",\n \"The filtered peak values are classified in an order of energy magnitude and filtered to the predetermined number smaller than 30(step S120).\",\n \"Each of the filtered peak values of the frequency data includes a formant frequency of a voice signal of a corresponding frame.\",\n \"A pitch is detected from a frequency area to obtain a fundamental frequency (step S122).\",\n \"The fundamental frequency is sound/image converted by a sound/image conversion algorithm to obtain a visible frequency (step S124).\",\n \"The sound/image conversion algorithm in step S124 is defined by the following equation 1.Equation 1 F = F l × 2 x * ( 1 ) B F ∝ x x = log 2 ( F i F l ) + C , where, F is a visible frequency to be obtained, F1 is a reference visible frequency, an integer party of x is an octave value, BF is a brightness of a color, x is a party down a decimal point and a sound field value in one octave, fi is a sampled reference audible frequency, f1 is a reference audible frequency, and C is a constant and a real number which ranges 0≦C≦1.For example, the reference audible frequency f1 is 329.6 Hz corresponding to ‘mi’ of 4 octave.\",\n \"The constant C is 0.29 by a reference frequency corresponding to ‘red’.\",\n \"When a minimal audible frequency f, and a minimal visible frequency F1 are set to 20 Hz and 350 Hz, respectively, a visible frequency is 441 THz, a brightness of a color to be obtained is 4.33(43.3%) proportional to the calculated x value according the equation 1.When a color corresponding to the visible frequency obtained in step S124 and image size information corresponding to energy of each frequency are formed as image data and the formed image data are provided to the central processing unit 602 (step S126), the central processing unit 602 converts image data into RGB data.\",\n \"The central processing unit 602 synthesizes the RGB data with another display data to generate image data.\",\n \"The image data are provided to an LCD driver 302 and are displayed through an LCD panel 304 (step S128).\",\n \"The central processing unit 602 controls the LCD panel 304 to display the image data in synchronism with an output of a voice call sound.\",\n \"FIG.\",\n \"10 is a view for showing a corresponding relationship of each piece of sound analyzed information and image information with respect to a voice call sound and an image display thereof.\",\n \"With reference to FIG.\",\n \"10, FIG.\",\n \"0.10a) shows a sampled voice signal, FIG.\",\n \"10b) shows a frequency spectrum of a fast Fourier transformed voice signal, and circle images corresponding to peak values of frequency data are displayed on a screen in FIG.\",\n \"10c).\",\n \"In FIG.\",\n \"10c), each size of circles corresponding to peak values of frequency data is proportioned to the energy of each frequency.\",\n \"A color of each circle is determined by the equation 1.Parts in which circles overlap with each other indicate that colors of each circle are mixed.\",\n \"A color corresponding to each audible frequency has a color shown in the color coordinate system of FIG.\",\n \"11 according to an octave and a musical scale.\",\n \"FIGS.\",\n \"12 through 18 are views for showing various image display statuses of a call sound according to an embodiment of the present invention.\",\n \"In FIGS.\",\n \"12 and 13, images corresponding to each sound frequency are arranged corresponding to a y axis of a screen and are displayed as areas proportional to energy amplitude of each sound frequency.\",\n \"In FIG.\",\n \"14, mixed colors of all frequencies are displayed from a center of a screen.\",\n \"The mixed colors are displayed to be radially progressed to an edge of the screen as time goes by.\",\n \"FIG.\",\n \"15 shows a combination of squares having different areas according to energy magnitude of each sound frequency to be displayed as an image forming a total of a screen by a Mondrian picture form.\",\n \"FIG.\",\n \"16 shows sounds of 4 octaves on a sound/color conversion system circle (radial circle coordinate which corresponds a sound to a color).\",\n \"The lowest octave is displayed at a square band of an outer most side.\",\n \"The highest octave is displayed at a square band of an inner most side.\",\n \"A musical scale of each sound is displayed as a sound corresponding to a square band.\",\n \"In FIG.\",\n \"17, an image such as a butterfly, a flower, and a star having a color corresponding to a musical scale of a sound to be displayed is displayed at a random coordinate.\",\n \"A previously displayed image does not disappear and the next displaying image overlaps with the previously displayed image.\",\n \"In FIG.\",\n \"18, an image of a regular triangle having a color corresponding to a musical scale of a sound.\",\n \"Furthermore, various forms such as a color bar form and a mosaic form can be used without departing from the spirit and scope of the invention.\",\n \"Therefore, a portable phone user takes pleasure in a bell sound not only aurally but also visibly by a color image which rhythmically changes on a liquid crystal panel according to the bell sound.\",\n \"3.Color Display of Bell Sound of Portable Phone According to Color Illuminating Unit A portable phone having a color liquid crystal panel was described in the embodiment.\",\n \"However, in a portable phone having a gradation display liquid crystal panel, the bell sound can be visibly displayed.\",\n \"That is, the portable phone includes a color illumination unit 306.The color illumination unit 306 includes illumination sources having at least two different colors.\",\n \"The color illumination unit 306 provides a color illumination to the liquid crystal panel 304.For example, the color illumination unit 306 outputs the color illumination by a combination of a red light emitting diode, a green light emitting diode, and a blue light emitting diode.\",\n \"Accordingly, in an embodiment of the present invention, in driving a color light emitting diode of the color illumination unit 306, in response to each gain information of RGB value corresponding to each sound obtained by the algorithm, a brightness of each color light emitting diode is controlled.\",\n \"Thus, in response to the bell sound, as shown in FIG.\",\n \"19, an illumination color is outputted to be rhythmically changed to 65000 colors on an LCD display window.\",\n \"4.Image Conversion Algorithm of Traffic Sound As shown in FIG.\",\n \"20, according to an image conversion display of the traffic sound, a called party's voice in a portable phone is outputted as an image and simultaneously, a feeling status and a purity according to the called party's voice analysis are expressed as characters.\",\n \"The vocoder 604 decodes data and generates ADPCM data.\",\n \"The ADPCM data are provided to a voice processor 202.The sound/image converter 606 samples output data of the vocoder 604 in frames to generate 1024 samples and converts them into PCM data (step S130).\",\n \"An average energy Ea ( E a = 1 N ∑ i = 1 N ( E si ) 2 ) by frames is obtained (step S132).\",\n \"Es is energy of each sample value, and N is the total sample number (1024) in a frame.\",\n \"It is judged whether an average energy Eaj of a current frame minus an average energy Ea(j−1) of a previous frame is greater than 6 db (step S134).\",\n \"When a traffic sound is present, an average energy of the initial 3 frames is obtained to calculate a reference energy Er.\",\n \"Then, an average energy AEk of the next 3 frames is obtained (step S136).\",\n \"Sample values of each frame are fast-Fourier-transformed (step S138).\",\n \"A pitch, that is, a fundamental frequency ff is obtained based on Fourier-transformed frequency data using a known algorithm which obtains a pitch (step S140).\",\n \"4-1.Voice Image Formation A fundamental frequency ff, is extracted.\",\n \"A circle proportional to an amplitude of energy Eff of the extracted fundamental frequency ff is arranged at a center of a voice screen area.\",\n \"A waveform at a time axis of a voice overlaps a circumference of a circle to output a voice image.\",\n \"A base color of a screen is displayed as a color corresponding to the fundamental frequency ff.\",\n \"The color corresponding to the fundamental frequency ff is acquired using the equation 1.As mentioned above, traffic sound image data are formed (step S142).\",\n \"4-2.Feeling Index Image Formation When a person is excited, the person speaks in a loud voice according to a feeling index to thereby change a volume and the fundamental frequency.\",\n \"According to an embodiment of the present invention, the feeling index is calculated based on energy values and fundamental frequencies by frames.\",\n \"A reference frequency ff is determined using fundamental frequencies ff1, ff2, and ff3 of initial 3 frames.\",\n \"The reference frequency fr, is determined as an average fundamental frequency ffa of the initial 3 frames.\",\n \"The average fundamental frequency is determined as an average of two fundamental frequencies except a fundamental frequency having a maximal value among the fundamental frequencies of the initial 3 frames.\",\n \"When the determined average reference frequency is within the range from 80 Hz to 600 Hz, the determined average reference frequency is determined as a reference frequency.\",\n \"When the determined average reference frequency is beyond the range from 80 Hz to 600 Hz, the greater one of the two fundamental frequency is determined as the reference frequency fr.\",\n \"When the reference energy Er and the reference frequency fr are determined, an average energy AEk and an average fundamental frequency fak are obtained in the next 3 frames.\",\n \"A change rate (E rate) of an average energy obtained with respect to the reference energy Er is obtained, that is, E rate=AEk/Er.\",\n \"A change rate (f rate) of an average fundamental frequency obtained with respec to the reference frequency fr is obtained, that is, f rate=fak/fr.\",\n \"Feeling indexes of 10 stages are determined as shown in table 2 according to the change rate (E rate) of an average energy and the change rate (f rate) of an average fundamental frequency.\",\n \"When the feeling indexes are determined, as shown in FIG.\",\n \"21, character data corresponding to the feeling indexes are read to form an image (step S144).\",\n \"A kind of feeling index character can be determined by a user's selection.\",\n \"TABLE 2 FREQUENCY FEELING INDEX ENERGY CHANGE RATE CHANGE RATE 10 GREATER THAN 13 1.2˜3 9 11˜13 8 9˜11 7 7˜9 6 6˜7 0.8˜2 5 5˜6 4 4˜5 3 3˜4 2 0.7˜3 0.7˜1.5 1 0˜0.7 0.1˜1.2 4-3.Purity Index Image Formation In an embodiment of the present invention, the purity of a sound is displayed proportional to a chroma of a color.\",\n \"That is, a clear sound is displayed close to one fundamental frequency.\",\n \"An impure sound has a fundamental frequency and a harmonic frequency.\",\n \"Accordingly, in a frequency spectrum per frame, the clear sound has a sampled low frequency.\",\n \"The impure sound has a sampled high frequency.\",\n \"The number of corresponding colors becomes larger in each frequency by the equation 1.The more a mixture thereof, the smaller a chroma of a mixed color becomes.\",\n \"RGB data f1 (RGB), f2 (RGB), f3 (RGB), .\",\n \".\",\n \".\",\n \"of sampled peak values per frame are obtained by the equation 1.RGB data are multiplied by energy value of each peak as a gain to obtain a sum of RGB so that a total RGB value is obtained every frame.\",\n \"T(RGB)=e1f1+e2f2+e3f3+... Energy value is regulated as a value from 0 to 1.A total RGB value has chroma information of a corresponding frame indicating all colors corresponding to each frequency are mixed with each other.\",\n \"Accordingly, a maximal value (MAX) and a minimal value (MIN) among each R value, each G value, and each B value are obtained from the total RGB value.\",\n \"A sum (SUM) and a difference (DIF) of the maximal value (MAX) and the minimal value (MIN) are obtained.\",\n \"The sum (SUM) is divided by 510 to obtain a luminance (LUM).\",\n \"When the maximal value (MAX) and the minimal value (MIN) are identical with each other, a chroma is 0.When the luminance (LUM) is equal to or greater than 0.5, the chroma is determined by a value which is obtained by dividing the difference (DIF) by the sum (SUM).\",\n \"When the luminance (LUM) is less than 0.5, the chroma is determined by a value which is obtained by dividing the difference (DIF) by (510−SUM).\",\n \"As shown in table 3, purity indexes of 10 grades are expressed corresponding to chromas of 10 grades.\",\n \"TABLE 3 CHROMA PURITY DEGREE INDEX 0˜0.1 −2 0.1˜0.2 −1 0.2˜0.3 0 0.3˜0.4 1 0.4˜0.5 2 0.5˜0.6 3 0.6˜0.7 4 0.7˜0.8 5 0.8˜0.9 6 0.9˜1.0 7 When a purity index is determined, as shown in FIG.\",\n \"22, character data corresponding to the purity index are read to form purity degree image data (step S146).\",\n \"A kind of a character to be displayed can be determined by a user's selection.\",\n \"When a traffic sound image, a feeling index image, and a purity degree image are formed, the sound/image converter 606 provides the traffic sound image, the feeling index image, and the purity degree image to the central processing unit 602 (step S148).\",\n \"The central processing unit 602 synthesizes the traffic sound image, the feeling index image, and the purity degree image from the sound/image converter 606 to form one display screen.\",\n \"The one display screen is displayed on the liquid crystal panel as shown in FIGS.\",\n \"20 and 21.With reference to FIG.\",\n \"23, in a traffic sound display area, a called party' voice waveform having a circle shape is displayed on bright violet corresponding to a fundamental frequency (5 octave B musical scale) ground.\",\n \"A character corresponding to a feeling index 7 is displayed in a feeling index display area.\",\n \"A character corresponding to a purity degree 0 is displayed in a purity degree index area.\",\n \"With reference to FIG.\",\n \"24, in a traffic sound display area, a called party' voice waveform having a circle shape is displayed on dark violet corresponding to a fundamental frequency (3 octave B musical scale) ground.\",\n \"A character corresponding to a feeling index 7 is displayed in a feeling index display area.\",\n \"A character corresponding to a purity degree +2 is displayed in a purity degree index area.\",\n \"A sound waveform of FIG.\",\n \"24 is greater than that of FIG.\",\n \"23.Accordingly, a user can visibly see the called party's volume magnitude.\",\n \"Luminances of FIGS.\",\n \"24 and 25 are different from each other according to a difference of the 5 octave and 3 octave.\",\n \"The user can recognize whether the call party's current feeling is in an excited or quiet state.\",\n \"The user can recognize whether the called party's voice is clear or impure through a character and an index.\",\n \"5.Another Example of an Image Conversion of a Traffic Sound Formant frequency information and pitch frequency information extracted in a decoding operation of the vocoder 104 are provided to a sound/image converter 606.A color corresponding to the formant frequency information and pitch frequency information is obtained by the equation 1.In the case, it is unnecessary for the sound/image converter 606 to perform a fast-Fourier transformation operation.\",\n \"Further, a sound/image converter transplants a program in a controller and forms it with a software.\",\n \"The sound/image converter can be integrated to a custom semiconductor.\",\n \"A sound/image converter and a voice synthesizer can be hardware embodied by a custom semiconductor.\",\n \"As mentioned above, according to the present invention, since a call sound of a portable phone, that is, a bell sound is audibly outputted and is visibly displayed on a screen, a newly added function is provided to the portable phone to improve a marketability.\",\n \"Also, since a called party's voice is displayed as an image, the called party's feeling and voice purity degree are displayed, additional pleasure is provided to the portable phone user, and marketability of the product is improved.\",\n \"While this invention has been particularly shown and described with reference to particular embodiments thereof, it will be understood by those skilled in the art that various changes in form and details may be effected therein without departing from the spirit and scope of the invention as defined by the appended claims.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473872"}}},{"rowIdx":759,"cells":{"text":{"kind":"list like","value":[["Process for producing foarmed articles and the foamed article","It is an object of the present invention to provide a method for manufacturing a high-quality foam that can be produced stably on an industrial scale and that affords excellent thermal insulation as a foam with thin cell membranes, which method ensures uniform molten mixture fluidity, which is an essential condition for achieving consistent foam quality, in the manufacture of a foam by mixing starch as one molten polymer compound with another polymer compound having a different melt viscosity, and to provide a foam manufactured by this method.","Water is added to and mixed with a plant-derived fibrous material and starch to manufacture a foam of a plastic resin containing as compositional components starch and a plant-derived fibrous material such as pulverized paper, pulverized wood, or the like.","A foam is manufactured by a method comprising a step of producing pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination, and the step of foaming the pellets by mixing them with a plastic resin, and included in between these two steps is a step of drying the pellets at a temperature not higher than the starch agglutination temperature."],["1.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing the pellets with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature.","2.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower.","3.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component.","4.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component.","5.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the pulverized paper is made from high-grade waste paper, or from printing scraps or waste of postal cards, stamps such as postage stamps, currency, and so forth.","6.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the pulverized paper is made from high-grade waste paper, or from printing scraps or waste of postal cards, stamps such as postage stamps, currency, and so forth.","7.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized wood as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component.","8.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, the plant-derived fibrous material comprising pulverized wood as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component.","9.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized wood as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the pulverized wood is of reed or the like.","10.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, the plant-derived fibrous material comprising pulverized wood as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the pulverized wood is of reed or the like.","11.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing the pellets with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.","12.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.","13.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.","14.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.","15.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the pulverized paper is made from high-grade waste paper, or from printing scraps or waste of postal cards, stamps such as postage stamps, currency, and so forth, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.","16.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the pulverized paper is made from high-grade waste paper, or from printing scraps or waste of postal cards, stamps such as postage stamps, currency, and so forth, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.","17.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized wood as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.","18.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, the plant-derived fibrous material comprising pulverized wood as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.","19.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized wood as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the pulverized wood is of reed or the like, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.","20.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, the plant-derived fibrous material comprising pulverized wood as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the pulverized wood is of reed or the like, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.","21.A foam, manufactured by the foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing the pellets with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.","22.A foam, manufactured by the foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.","23.A foam, manufactured by the foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.","24.A foam, manufactured by the foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.","25.A foam, manufactured by the foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the pulverized paper is made from high-grade waste paper, or from printing scraps or waste of postal cards, stamps such as postage stamps, currency, and so forth, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.","26.A foam, manufactured by the foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the pulverized paper is made from high-grade waste paper, or from printing scraps or waste of postal cards, stamps such as postage stamps, currency, and so forth, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.","27.A foam, manufactured by the foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized wood as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.","28.A foam, manufactured by the foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, the plant-derived fibrous material comprising pulverized wood as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.","29.A foam, manufactured by the foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized wood as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the pulverized wood is of reed or the like, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.","30.A foam, manufactured by the foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, the plant-derived fibrous material comprising pulverized wood as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the pulverized wood is of reed or the like, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %."],["<SOH> BACKGROUND ART <EOH>There have been countless attempts at manufacturing a foam by mixing starch with another plant-derived material or with a plastic resin that would pose no danger of emitting dioxins if incinerated after use, for example, and at utilizing such foams as thermal insulation materials or as cushioning materials or other such shipping and packaging materials, and a number of methods for manufacturing foams have been disclosed.","Nevertheless, based on an examination of this disclosed information, it appears that no known technique has been established relating to the problem the present invention is intended to solve.","To clarify the problem the present invention is intended to solve, general concepts, beliefs, related examples, and so forth were examined in the technological field to which the present invention is related.","It is commonly held that when steam or the like is used as a foaming gas, the material is discharged under pressure from an extrusion molding machine, and reduced-pressure foaming is performed, for instance, a polymer raw material that becomes fluid when the temperature is raised will flow as shown in FIG.","6A , with symmetrical flow lines around the center line in the drawing, when the pressure is relatively low, and as the pressure is gradually raised, slip is observed between the material and the walls.","At even higher pressures the flow lines begin to be disarrayed, the material is extruded in a spiral shape, and this results in a lack of uniformity in the material.","What happens here is called stick and flow, in which the flow of the material comes to a halt (sticking), after which the material starts sliding again (slipping).","When this occurs, complete and intermittent slipping over the entire surface of the slip cone shown in FIG.","6B has been reported for high density polyethylene and other such polymer materials, and a similar phenomenon is seen not just with polymer compounds but also in the extrusion of bismuth-tellurium and other such metal materials that are thermoelectric semiconductors, for example.","This is manifested as cracks in the extruded semiconductor.","Various apparatus improvements have been attempted from a mechanical aspect in an effort to eliminate this problem, but the fact is that the fluid properties of polymer compounds are still not fully understood by researchers.","The materials that are the subject of the present invention are polymer compounds, and it is necessary to understand this phenomenon.","Since the present invention involves a plurality of polymer compounds, however, it is uncertain whether the phenomenon witnessed with a single compound such as polyethylene would be the same, and the problem is even more complicated because we are dealing with a mixture in which the temperature dependency of viscosity and the thermal deformation temperature characteristics vary with the different raw material polymer compounds, which probably makes it even more difficult to understand the situation in terms of fluid dynamics or tribology.","The present invention relates to a field of technology in which such scientific understanding is not necessarily perfect, but at the very least, of course, we must know something about the material characteristics of the raw material polymer compounds being used.","Let us now give a brief summary of the characteristics of a polypropylene resin, as an example of a plastic resin, and starch, which are believed to be the main contributors to fluidity out of the polymer compound materials used in the present invention.","Starch is usually in the form of macromolecular spherulites of a mixture of two homologues with different structures, amylose and amylopectin.","The size thereof can vary greatly with the type of starch, but the diameter is said to range from 1 to 20 microns, and the molecular weight up to a few thousand.","Amylose is a linear polymer to which D-glucose units are connected by α-1,4 linkages, whereas amylopectin is a branched polymer that includes α-1,6 linkages in addition to α-1,4 linkages, with branching occurring at these sites and the branches connected again with α-1,4 linkages.","When starch granules are suspended in water and heated to a certain temperature or higher, the starch granules undergo irreversible swelling and amylose is eluted.","This process is called agglutination, and this temperature is referred to as the agglutination temperature.","FIG.","7 shows published heating and viscosity curves for various kinds of starch (amylograms).","The agglutination temperature ranges from 60 to 90° C., and cornstarch is what is cited in the embodiments and so forth of the present invention.","FIG.","8 also gives published information, and is a comparison of the viscosity behavior of ordinary wheat resulting from agglutination, by variety of glutinous wheat (which has a relatively high amylopectin content), versus that of cornstarch.","For this comparison, the temperature of each starch was raised from 34° C. to 94° C. at a rate of 5° C. per minute, after which the starch was cooled at the same rate of 5° C. per minute and its viscosity was measured.","Of these varieties, Norin No.","61 had the same amylose content of 31.8% as ordinary wheat, and its agglutination temperature was close to that of wheat as shown in FIG.","8 .","The waxy cornstarch used as a control also has a relatively low agglutination temperature, similar to what is shown in FIG.","7 .","The purpose of providing FIGS.","7 and 8 here is to show the agglutination temperature of the starch that is one of the polymer compound materials used in the present invention in relation to the fluidity of these materials, and demonstrate that there is a rapid increase in viscosity as soon as the agglutination temperature is attained.","The polypropylene resin will now be discussed.","Polypropylene is a crystalline thermoplastic resin made by the polymerization of propylene (C 3 H 6 ), and is hard and tough, with good resistance to moisture, oils, and solvents.","Its heat resistance temperature is said to be 170° C. or lower.","Also, there is almost no possibility that a linear polymer compound will undergo cyclization during combustion, and these resins, along with polyethylene and the like, are called polyolefin resins and said to be resins that do not produce dioxins or other harmful substances.","According to disclosed reference materials from polypropylene manufacturers and so forth, the thermal deformation temperature of polypropylene varies somewhat with the structure of the compound, but is said to be roughly 100 to 120° C. FIG.","9 shows an example of the elongation viscosity of the linear (without branches) polypropylene resin used in the examples and so forth of the present invention, and it can be seen that the elongation viscosity at 180° C. quickly disappears in just a few seconds.","The reason a foam of a high expansion ratio cannot be produced from a linear polypropylene resin is said to be that the elongation viscosity drops off so quickly and the melt tension is low, and most polypropylene manufacturers produce polypropylene resins having a branched structure by subjecting a linear polypropylene to a special process, and then make a foam with a relatively high expansion ratio from this polypropylene resin with improved elongation viscosity and melt tension.","Higher cost is at the present time inevitable with a branched polypropylene because a linear polypropylene must be subjected to a post-treatment step such as electron beam irradiation.","When an attempt is made to produce a foam by mixing and melting a starch and a plastic resin such as polypropylene, the difference between the temperature dependence of the viscosity behavior and the melt temperature of the polymer compounds results in complex fluidity of the mixed materials in the mixing, melting, and extrusion steps during manufacture, and it is not hard to imagine the effect that this has on discharge uniformity.","Japanese Laid-Open Patent Application H9-111029 discusses a method for manufacturing a loose polypropylene foam by mixing a polypropylene resin and a starch-based additive into a plant-derived foaming agent such as nonfat powdered milk or bean-curd lees, putting this raw material mixture into an extruder comprising a high-temperature heated cylinder, and feeding in water.","This publication also discusses a method in which a plastic component, plant-derived foaming agent, and starch-based additive are simply mixed.","The inventors of the present invention conducted manufacturing tests in which they used an actual foam manufacturing apparatus similar to that in the disclosed information, used as their raw materials a plastic resin, starch, and pulverized waste paper (as the plant-derived fibrous material), and varied the amount of feed water (the foaming agent), the mixing time and temperature in the manufacturing process, the conditions of the material discharge orifice, and so forth.","As a result, with a simple mixture of raw materials as in the disclosed information, perhaps because variance in fluidity could not be suppressed, there was considerably fluctuation in the discharge amount per hour, and there was also variance in the foaming state, so it was found that a foam of stable quality could not be manufactured.","The specific manufacturing method and steps here are illustrated in FIG.","10 .","Pulverized paper (such as waste paper) was used as the plant-derived fibrous material, but first the material was finely pulverized in a mixer/cutter type of pulverizer and then in a ball mill type of pulverizer, this pulverized material was sifted through an industrial sieve to obtain a waste paper powder with particles averaging 20 to 30 microns in size, and this powder was used as the waste paper raw material.","This waste paper raw material was then mixed with a polypropylene resin to obtain a raw material mixture, to which cornstarch was further added and mixed, and these components were put in a mixing machine and mixed.","The sequence of this mixing was also varied so that the waste paper raw material was first mixed with the cornstarch to obtain a raw material mixture, to which the polypropylene resin was added and mixed in a mixing machine.","The mixing was also performed at various blend ratios, including those given in the disclosed information.","This kneaded raw material was put into,a biaxial extruder and foamed.","Specifically, the biaxial extruder shown in FIG.","11 comprises a raw material inlet 1 , a water inlet 2 , cylinders C 1 to C 5 , heaters 3 attached to the various cylinders, a raw material extrusion die (spinneret) 4 , and an extrusion opening 5 .","In manufacturing tests, the settings of the cylinder and extrusion die temperatures was suitably varied.","The test was conducted with the water supply varied between 10 and 30 liters per hour.","To evaluate the foam state and foam variance, a string-like piece of foam extruded and foamed from an extrusion die with a diameter of 2 mm was cut up by an automatic cutter set to a specific rotating speed, 50 pieces were randomly sampled from among the total of 4200 pieces of cut foam produced in one minute, whose combined length was 120 m, the length and diameter of the cut foam pieces were measured, and the extent of foaming was visually evaluated on a ten-point scale.","The mixing ratios of the materials for which the results are shown in FIG.","14 were 25% (weight ratio) pulverized waste paper, 30% starch, and 45% polypropylene resin.","As indicated by these results, the foam manufactured by the mixing process described above exhibited considerable variance in all measurement categories.","The length of cushioning material cut at regular time intervals varied from a minimum of 1.9 cm to a maximum of 8.5 cm.","The most serious problem was variance in the extent of foaming.","The ten-point scale assumes 10 to be the ideal foaming state, but because there was variance in the foaming, the cushioning material varied from thin to thick.","This indicates nothing other than a change over time in the fluidity of the raw material mixture during mixing and melting inside the cylinders.","This is similar to the stick and flow phenomenon described above, but it is unclear whether it occurred because a slip cone was formed, or just because of local variance in the fluidity of the molten mixture.","Admixed in this molten mixture was pulverized waste paper, whose characteristics remained almost unchanged over the temperature range in this process.","This pulverized paper consists of a fibrous material with a microscopic length of anywhere from a few microns to about 100 microns, which is stirred and mixed in the biaxial extruder, and it seems that nuclear cells that will serve as the nuclei for cells are produced here and there in an agitated fluid in which polymer compounds of different viscosity are admixed.","In this situation it is doubtful whether a nice slip cone is being formed as in the above-mentioned reports on polyethylene resins.","On the contrary, it seems likely that this nucleus formation is occurring as a result of local variance in the fluidity of the molten mixture.","At this point we will briefly discuss the correlation between the viscosity of the above-mentioned starch and polypropylene resin and the fluidity of the mixture of polymer compounds, since such a discussion will probably further clarify the points involved in the problems which the present invention is intended to solve and the means used to solve the problems.","A capillary type of viscosity measurement apparatus is the most commonly used in the measurement of the viscosity of polymers.","A sample is extruded from a capillary tube by applying pressure, and the viscosity is measured by measuring the difference in the applied pressure at a constant flow rate per unit of time.","We will let L be the length of the capillary tube, and r be its radius.","The change over time in the amount of sample flowing out of the capillary tube is found for when pressure is applied such that the sample contained in the reservoir flows at a specific rate, and this is termed Q (cm 3 /sec).","A relationship of in-line-formulae description=\"In-line Formulae\" end=\"lead\"?","γ=4 Q/πr 3 in-line-formulae description=\"In-line Formulae\" end=\"tail\"?","exists between the shear rate γ (sec −1 ) and the sample flowing through the capillary tube, and the shear stress τ (N/cm 2 ) on the inner walls of the capillary tube is expressed as τ=rP/2L.","The viscosity μ (N·sec/cm 2 ) is defined from the shear rate y and the shear stress τ as: in-line-formulae description=\"In-line Formulae\" end=\"lead\"?","μ=τ/γ.","in-line-formulae description=\"In-line Formulae\" end=\"tail\"?","If we assume that a starch and a polypropylene resin are mixed and the thermal characteristics of each are independently manifested, the intrinsic viscosity should be exhibited in the starch at the agglutination temperature and in the polypropylene resin at the thermal deformation temperature, with the viscosity subsequently varying according to temperature changes.","For instance, if the agglutination temperature of the starch is reached before the thermal deformation temperature is reached, the viscosity of the starch increases suddenly, and the entire system is governed by the viscosity of the starch.","If the thermal deformation temperature of the polypropylene resin is never reached, the polypropylene might have some kind of effect as an admixture, but the viscosity μ of the system will be relatively stable.","In other words, this is a state in which the fluidity of the system is governed by the viscosity of the starch, even though the system is a mixture.","However, this never happens for the purposes of manufacturing a foam.","Still, if the temperature continued to rise, it would eventually reach the thermal deformation temperature of the polypropylene resin, resulting in a fluid having both the viscosity of polypropylene and the viscosity of the starch, which varies with time and temperature.","It is very difficult to know what the mixing state is in a mixed fluid such as this.","Measurement was made at a certain specific temperature with the above-mentioned viscosity measurement method, but since the mixing is not completely uniform in a mixing state of different viscosities, the local shear rate γ should vary.","Furthermore, the shear stress τ on the inner walls of the capillary tube also varies according to the local mixing state.","In other words, the viscosity μ of a mixture of polymer compounds changes with how uniform the mixing is, and if we look at the viscosity definition of the above formula, we can see that there is extreme instability in which both the denominator and the numerator vary.","Put another way, there is no viscosity that governs the mixture as a system, and instead the system is in a state of constant change.","Japanese Laid-Open Patent Application 2000-143869, which is another source of disclosed information, discloses a technique of manufacturing a foam by pulverizing a plastic laminate and mixing with polypropylene, cornstarch, and calcium carbonate.","With this technique, the foaming expansion ratio is about 10 times with just the resin that is admixed in the laminate paper, and the admixture of another plastic is proposed in order to increase the foaming expansion ratio.","Close examination of this technique reveals that, as seen in the embodiments and examples of the invention, a foamable composition is made up of plastic from pulverized laminate paper and added calcium carbonate.","Cornstarch and the paper component of plastic laminate paper are added as fillers, the components are stirred and mixed, put into the hopper of an extrusion molding machine, kneaded, heated, melted, and pressurized while water is supplied, and then discharged into the air and expanded to obtain a continuous foam.","One way to increase the foaming expansion ratio is to add a plastic material that is miscible with the plastic component of the plastic laminate paper.","After the plastic material, organic plant component, and inorganic component are kneaded, water is added as a foaming agent, and it is considered important that the various components be uniformly dispersed if a uniform foam is to be obtained.","To this end, it is preferable if discarded plastic laminate paper is pulverized and made into pellets in a pelletizer, to which are added the required amounts of plastic, organic plant component, and inorganic component, the components are stirred and kneaded in a mixer, and the resulting composition is supplied to an extrusion molding machine.","With this technique, the manufacturing process is characterized in that the method for mixing the materials involves merely mixing the plastic component, plant component (such as cornstarch or paper), and inorganic component.","Even with this disclosed technique, there is no special technical description of the above mixing step other than to say that the goal is to obtain a uniform form by merely subjecting raw materials of different melting temperatures and viscosities to a kneading step, and achieving the uniform dispersion of the various components.","It is obvious that this disclosed information does not solve the problem related to viscosity and fluidity encountered by the inventors of the present invention.","A foam is basically a resource-conserving material in that the amount of materials consumed is extremely small.","Foams have excellent adiabatic properties and impact resistance, and are lightweight, which means that they can be used in a wide variety of situations, including applications as packaging materials, and for this reason resin manufacturers and others have conducted intensive research into foams.","The flip-side of being lightweight is that foams also have a large volume, so their development requires adequate consideration of their material characteristics as related to recyclability and disposal.","As already touched upon in prior art, if a foam could be made from pulverized waste paper or another plant-derived fibrous component or a starch or the like and, if needed, a plastic resin or the like that has been confirmed to be safe to humans and the environment, this would have considerable social significance as well, in view of the high cost of developing new resins and so forth.","The present invention solves the problems with viscosity and fluidity inevitably encountered with mixtures of a plurality of different types of polymer compound in the manufacture of a foam by mixing a plastic resin with the above-mentioned natural materials, and establishes a method for manufacturing a foam of uniform quality, with which stable manufacture on an industrial scale is possible."],["<SOH> BRIEF DESCRIPTION OF THE DRAWINGS <EOH>FIG.","1 is a concept diagram of the manufacturing method of the present invention; FIG.","2 is a cross sectional micrograph of the present invention; FIG.","3 is a cross sectional micrograph of the present invention; FIG.","4 is a micrograph of the interior of the foam of the present invention; FIG.","5 is a data table of the results of foam manufacture in the present invention; FIG.","6A is a flow state diagram of when the pressure is relatively low in the reservoir in prior art, and FIG.","6B is a flow state diagram of slip cone generation when the pressure is relatively high in prior art; FIG.","7 is a graph of the viscosity of various kinds of starch in prior art; FIG.","8 is a graph of starch viscosity characteristics in prior art; FIG.","9 is a graph of elongation viscosity characteristics in prior art; FIG.","10 is a flow chart of a foam manufacturing method in prior art; FIG.","11 is a simplified diagram of the main parts of a foam manufacturing apparatus in prior art; FIG.","12 is a cut-away oblique view of the raw material extrusion die in the manufacture of a foam board in prior art; FIG.","13 is a manufacturing state diagram of a foam board in prior art; and FIG.","14 is a data table of the results of foam manufacture in prior art.","detailed-description description=\"Detailed Description\" end=\"lead\"?"],["TECHNICAL FIELD This invention relates to a method for manufacturing a high-quality foam that can be produced stably on an industrial scale and that affords excellent thermal insulation as a foam with thin cell membranes, which method ensures uniform molten mixture fluidity, that is an essential condition for achieving consistent foam quality, in the manufacture of a foam by mixing starch as one molten polymer compound with another polymer compound having a different melt viscosity, and to a foam manufactured by this method.","BACKGROUND ART There have been countless attempts at manufacturing a foam by mixing starch with another plant-derived material or with a plastic resin that would pose no danger of emitting dioxins if incinerated after use, for example, and at utilizing such foams as thermal insulation materials or as cushioning materials or other such shipping and packaging materials, and a number of methods for manufacturing foams have been disclosed.","Nevertheless, based on an examination of this disclosed information, it appears that no known technique has been established relating to the problem the present invention is intended to solve.","To clarify the problem the present invention is intended to solve, general concepts, beliefs, related examples, and so forth were examined in the technological field to which the present invention is related.","It is commonly held that when steam or the like is used as a foaming gas, the material is discharged under pressure from an extrusion molding machine, and reduced-pressure foaming is performed, for instance, a polymer raw material that becomes fluid when the temperature is raised will flow as shown in FIG.","6A, with symmetrical flow lines around the center line in the drawing, when the pressure is relatively low, and as the pressure is gradually raised, slip is observed between the material and the walls.","At even higher pressures the flow lines begin to be disarrayed, the material is extruded in a spiral shape, and this results in a lack of uniformity in the material.","What happens here is called stick and flow, in which the flow of the material comes to a halt (sticking), after which the material starts sliding again (slipping).","When this occurs, complete and intermittent slipping over the entire surface of the slip cone shown in FIG.","6B has been reported for high density polyethylene and other such polymer materials, and a similar phenomenon is seen not just with polymer compounds but also in the extrusion of bismuth-tellurium and other such metal materials that are thermoelectric semiconductors, for example.","This is manifested as cracks in the extruded semiconductor.","Various apparatus improvements have been attempted from a mechanical aspect in an effort to eliminate this problem, but the fact is that the fluid properties of polymer compounds are still not fully understood by researchers.","The materials that are the subject of the present invention are polymer compounds, and it is necessary to understand this phenomenon.","Since the present invention involves a plurality of polymer compounds, however, it is uncertain whether the phenomenon witnessed with a single compound such as polyethylene would be the same, and the problem is even more complicated because we are dealing with a mixture in which the temperature dependency of viscosity and the thermal deformation temperature characteristics vary with the different raw material polymer compounds, which probably makes it even more difficult to understand the situation in terms of fluid dynamics or tribology.","The present invention relates to a field of technology in which such scientific understanding is not necessarily perfect, but at the very least, of course, we must know something about the material characteristics of the raw material polymer compounds being used.","Let us now give a brief summary of the characteristics of a polypropylene resin, as an example of a plastic resin, and starch, which are believed to be the main contributors to fluidity out of the polymer compound materials used in the present invention.","Starch is usually in the form of macromolecular spherulites of a mixture of two homologues with different structures, amylose and amylopectin.","The size thereof can vary greatly with the type of starch, but the diameter is said to range from 1 to 20 microns, and the molecular weight up to a few thousand.","Amylose is a linear polymer to which D-glucose units are connected by α-1,4 linkages, whereas amylopectin is a branched polymer that includes α-1,6 linkages in addition to α-1,4 linkages, with branching occurring at these sites and the branches connected again with α-1,4 linkages.","When starch granules are suspended in water and heated to a certain temperature or higher, the starch granules undergo irreversible swelling and amylose is eluted.","This process is called agglutination, and this temperature is referred to as the agglutination temperature.","FIG.","7 shows published heating and viscosity curves for various kinds of starch (amylograms).","The agglutination temperature ranges from 60 to 90° C., and cornstarch is what is cited in the embodiments and so forth of the present invention.","FIG.","8 also gives published information, and is a comparison of the viscosity behavior of ordinary wheat resulting from agglutination, by variety of glutinous wheat (which has a relatively high amylopectin content), versus that of cornstarch.","For this comparison, the temperature of each starch was raised from 34° C. to 94° C. at a rate of 5° C. per minute, after which the starch was cooled at the same rate of 5° C. per minute and its viscosity was measured.","Of these varieties, Norin No.","61 had the same amylose content of 31.8% as ordinary wheat, and its agglutination temperature was close to that of wheat as shown in FIG.","8.The waxy cornstarch used as a control also has a relatively low agglutination temperature, similar to what is shown in FIG.","7.The purpose of providing FIGS.","7 and 8 here is to show the agglutination temperature of the starch that is one of the polymer compound materials used in the present invention in relation to the fluidity of these materials, and demonstrate that there is a rapid increase in viscosity as soon as the agglutination temperature is attained.","The polypropylene resin will now be discussed.","Polypropylene is a crystalline thermoplastic resin made by the polymerization of propylene (C3H6), and is hard and tough, with good resistance to moisture, oils, and solvents.","Its heat resistance temperature is said to be 170° C. or lower.","Also, there is almost no possibility that a linear polymer compound will undergo cyclization during combustion, and these resins, along with polyethylene and the like, are called polyolefin resins and said to be resins that do not produce dioxins or other harmful substances.","According to disclosed reference materials from polypropylene manufacturers and so forth, the thermal deformation temperature of polypropylene varies somewhat with the structure of the compound, but is said to be roughly 100 to 120° C. FIG.","9 shows an example of the elongation viscosity of the linear (without branches) polypropylene resin used in the examples and so forth of the present invention, and it can be seen that the elongation viscosity at 180° C. quickly disappears in just a few seconds.","The reason a foam of a high expansion ratio cannot be produced from a linear polypropylene resin is said to be that the elongation viscosity drops off so quickly and the melt tension is low, and most polypropylene manufacturers produce polypropylene resins having a branched structure by subjecting a linear polypropylene to a special process, and then make a foam with a relatively high expansion ratio from this polypropylene resin with improved elongation viscosity and melt tension.","Higher cost is at the present time inevitable with a branched polypropylene because a linear polypropylene must be subjected to a post-treatment step such as electron beam irradiation.","When an attempt is made to produce a foam by mixing and melting a starch and a plastic resin such as polypropylene, the difference between the temperature dependence of the viscosity behavior and the melt temperature of the polymer compounds results in complex fluidity of the mixed materials in the mixing, melting, and extrusion steps during manufacture, and it is not hard to imagine the effect that this has on discharge uniformity.","Japanese Laid-Open Patent Application H9-111029 discusses a method for manufacturing a loose polypropylene foam by mixing a polypropylene resin and a starch-based additive into a plant-derived foaming agent such as nonfat powdered milk or bean-curd lees, putting this raw material mixture into an extruder comprising a high-temperature heated cylinder, and feeding in water.","This publication also discusses a method in which a plastic component, plant-derived foaming agent, and starch-based additive are simply mixed.","The inventors of the present invention conducted manufacturing tests in which they used an actual foam manufacturing apparatus similar to that in the disclosed information, used as their raw materials a plastic resin, starch, and pulverized waste paper (as the plant-derived fibrous material), and varied the amount of feed water (the foaming agent), the mixing time and temperature in the manufacturing process, the conditions of the material discharge orifice, and so forth.","As a result, with a simple mixture of raw materials as in the disclosed information, perhaps because variance in fluidity could not be suppressed, there was considerably fluctuation in the discharge amount per hour, and there was also variance in the foaming state, so it was found that a foam of stable quality could not be manufactured.","The specific manufacturing method and steps here are illustrated in FIG.","10.Pulverized paper (such as waste paper) was used as the plant-derived fibrous material, but first the material was finely pulverized in a mixer/cutter type of pulverizer and then in a ball mill type of pulverizer, this pulverized material was sifted through an industrial sieve to obtain a waste paper powder with particles averaging 20 to 30 microns in size, and this powder was used as the waste paper raw material.","This waste paper raw material was then mixed with a polypropylene resin to obtain a raw material mixture, to which cornstarch was further added and mixed, and these components were put in a mixing machine and mixed.","The sequence of this mixing was also varied so that the waste paper raw material was first mixed with the cornstarch to obtain a raw material mixture, to which the polypropylene resin was added and mixed in a mixing machine.","The mixing was also performed at various blend ratios, including those given in the disclosed information.","This kneaded raw material was put into,a biaxial extruder and foamed.","Specifically, the biaxial extruder shown in FIG.","11 comprises a raw material inlet 1, a water inlet 2, cylinders C1 to C5, heaters 3 attached to the various cylinders, a raw material extrusion die (spinneret) 4, and an extrusion opening 5.In manufacturing tests, the settings of the cylinder and extrusion die temperatures was suitably varied.","The test was conducted with the water supply varied between 10 and 30 liters per hour.","To evaluate the foam state and foam variance, a string-like piece of foam extruded and foamed from an extrusion die with a diameter of 2 mm was cut up by an automatic cutter set to a specific rotating speed, 50 pieces were randomly sampled from among the total of 4200 pieces of cut foam produced in one minute, whose combined length was 120 m, the length and diameter of the cut foam pieces were measured, and the extent of foaming was visually evaluated on a ten-point scale.","The mixing ratios of the materials for which the results are shown in FIG.","14 were 25% (weight ratio) pulverized waste paper, 30% starch, and 45% polypropylene resin.","As indicated by these results, the foam manufactured by the mixing process described above exhibited considerable variance in all measurement categories.","The length of cushioning material cut at regular time intervals varied from a minimum of 1.9 cm to a maximum of 8.5 cm.","The most serious problem was variance in the extent of foaming.","The ten-point scale assumes 10 to be the ideal foaming state, but because there was variance in the foaming, the cushioning material varied from thin to thick.","This indicates nothing other than a change over time in the fluidity of the raw material mixture during mixing and melting inside the cylinders.","This is similar to the stick and flow phenomenon described above, but it is unclear whether it occurred because a slip cone was formed, or just because of local variance in the fluidity of the molten mixture.","Admixed in this molten mixture was pulverized waste paper, whose characteristics remained almost unchanged over the temperature range in this process.","This pulverized paper consists of a fibrous material with a microscopic length of anywhere from a few microns to about 100 microns, which is stirred and mixed in the biaxial extruder, and it seems that nuclear cells that will serve as the nuclei for cells are produced here and there in an agitated fluid in which polymer compounds of different viscosity are admixed.","In this situation it is doubtful whether a nice slip cone is being formed as in the above-mentioned reports on polyethylene resins.","On the contrary, it seems likely that this nucleus formation is occurring as a result of local variance in the fluidity of the molten mixture.","At this point we will briefly discuss the correlation between the viscosity of the above-mentioned starch and polypropylene resin and the fluidity of the mixture of polymer compounds, since such a discussion will probably further clarify the points involved in the problems which the present invention is intended to solve and the means used to solve the problems.","A capillary type of viscosity measurement apparatus is the most commonly used in the measurement of the viscosity of polymers.","A sample is extruded from a capillary tube by applying pressure, and the viscosity is measured by measuring the difference in the applied pressure at a constant flow rate per unit of time.","We will let L be the length of the capillary tube, and r be its radius.","The change over time in the amount of sample flowing out of the capillary tube is found for when pressure is applied such that the sample contained in the reservoir flows at a specific rate, and this is termed Q (cm3/sec).","A relationship of γ=4Q/πr3 exists between the shear rate γ (sec−1) and the sample flowing through the capillary tube, and the shear stress τ (N/cm2) on the inner walls of the capillary tube is expressed as τ=rP/2L.","The viscosity μ (N·sec/cm2) is defined from the shear rate y and the shear stress τ as: μ=τ/γ.","If we assume that a starch and a polypropylene resin are mixed and the thermal characteristics of each are independently manifested, the intrinsic viscosity should be exhibited in the starch at the agglutination temperature and in the polypropylene resin at the thermal deformation temperature, with the viscosity subsequently varying according to temperature changes.","For instance, if the agglutination temperature of the starch is reached before the thermal deformation temperature is reached, the viscosity of the starch increases suddenly, and the entire system is governed by the viscosity of the starch.","If the thermal deformation temperature of the polypropylene resin is never reached, the polypropylene might have some kind of effect as an admixture, but the viscosity μ of the system will be relatively stable.","In other words, this is a state in which the fluidity of the system is governed by the viscosity of the starch, even though the system is a mixture.","However, this never happens for the purposes of manufacturing a foam.","Still, if the temperature continued to rise, it would eventually reach the thermal deformation temperature of the polypropylene resin, resulting in a fluid having both the viscosity of polypropylene and the viscosity of the starch, which varies with time and temperature.","It is very difficult to know what the mixing state is in a mixed fluid such as this.","Measurement was made at a certain specific temperature with the above-mentioned viscosity measurement method, but since the mixing is not completely uniform in a mixing state of different viscosities, the local shear rate γ should vary.","Furthermore, the shear stress τ on the inner walls of the capillary tube also varies according to the local mixing state.","In other words, the viscosity μ of a mixture of polymer compounds changes with how uniform the mixing is, and if we look at the viscosity definition of the above formula, we can see that there is extreme instability in which both the denominator and the numerator vary.","Put another way, there is no viscosity that governs the mixture as a system, and instead the system is in a state of constant change.","Japanese Laid-Open Patent Application 2000-143869, which is another source of disclosed information, discloses a technique of manufacturing a foam by pulverizing a plastic laminate and mixing with polypropylene, cornstarch, and calcium carbonate.","With this technique, the foaming expansion ratio is about 10 times with just the resin that is admixed in the laminate paper, and the admixture of another plastic is proposed in order to increase the foaming expansion ratio.","Close examination of this technique reveals that, as seen in the embodiments and examples of the invention, a foamable composition is made up of plastic from pulverized laminate paper and added calcium carbonate.","Cornstarch and the paper component of plastic laminate paper are added as fillers, the components are stirred and mixed, put into the hopper of an extrusion molding machine, kneaded, heated, melted, and pressurized while water is supplied, and then discharged into the air and expanded to obtain a continuous foam.","One way to increase the foaming expansion ratio is to add a plastic material that is miscible with the plastic component of the plastic laminate paper.","After the plastic material, organic plant component, and inorganic component are kneaded, water is added as a foaming agent, and it is considered important that the various components be uniformly dispersed if a uniform foam is to be obtained.","To this end, it is preferable if discarded plastic laminate paper is pulverized and made into pellets in a pelletizer, to which are added the required amounts of plastic, organic plant component, and inorganic component, the components are stirred and kneaded in a mixer, and the resulting composition is supplied to an extrusion molding machine.","With this technique, the manufacturing process is characterized in that the method for mixing the materials involves merely mixing the plastic component, plant component (such as cornstarch or paper), and inorganic component.","Even with this disclosed technique, there is no special technical description of the above mixing step other than to say that the goal is to obtain a uniform form by merely subjecting raw materials of different melting temperatures and viscosities to a kneading step, and achieving the uniform dispersion of the various components.","It is obvious that this disclosed information does not solve the problem related to viscosity and fluidity encountered by the inventors of the present invention.","A foam is basically a resource-conserving material in that the amount of materials consumed is extremely small.","Foams have excellent adiabatic properties and impact resistance, and are lightweight, which means that they can be used in a wide variety of situations, including applications as packaging materials, and for this reason resin manufacturers and others have conducted intensive research into foams.","The flip-side of being lightweight is that foams also have a large volume, so their development requires adequate consideration of their material characteristics as related to recyclability and disposal.","As already touched upon in prior art, if a foam could be made from pulverized waste paper or another plant-derived fibrous component or a starch or the like and, if needed, a plastic resin or the like that has been confirmed to be safe to humans and the environment, this would have considerable social significance as well, in view of the high cost of developing new resins and so forth.","The present invention solves the problems with viscosity and fluidity inevitably encountered with mixtures of a plurality of different types of polymer compound in the manufacture of a foam by mixing a plastic resin with the above-mentioned natural materials, and establishes a method for manufacturing a foam of uniform quality, with which stable manufacture on an industrial scale is possible.","DISCLOSURE OF THE INVENTION The present invention provides a high-quality foam that can be produced stably on an industrial scale and that affords excellent thermal insulation as a foam with thin cell membranes.","Also, efficiently utilizing a starch (a natural polymer compound) has great industrial significance in that even a linear plastic resin that is inexpensive and has low elongation viscosity can be used to manufacture a foam with a high foaming expansion ratio.","Furthermore, the present invention provides a novel way of thinking about the scientific phenomena of agglutination and mixed fluids of a plurality of polymer compounds.","Also, with the present invention, in the manufacture of a foam of a plastic resin containing as compositional components pulverized paper, pulverized wood, or another such plant-derived fibrous material and starch, there is a step of adding water to the plant-derived fibrous material and starch and mixing the components to manufacture pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination, and a step of foaming the pellets by mixing them with a plastic resin, and included in between these two steps is a step of drying the pellets at a temperature not higher than the starch agglutination temperature.","The result is that it is possible to manufacture a foam of uniform quality, and one that has a high foaming expansion ratio even though it is made from an inexpensive linear plastic resin with low elongation viscosity.","Also, with the present invention, the pellets are dried for a relatively long time at a low temperature of 50° C. or lower, the advantage of which is a higher foaming expansion ratio.","Also, with the present invention, in the manufacture of a foam of a plastic resin containing as compositional components pulverized paper, pulverized wood, or another such plant-derived fibrous material and starch, there is a step of adding water to the plant-derived fibrous material and starch and mixing the components to manufacture pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination, and a step of foaming the pellets by mixing them with a plastic resin, and included in between these two steps is a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, or the pulverized paper being made from high-grade waste paper or from printing scraps or waste of postal cards, stamps such as postage stamps, currency, and so forth.","The result is that it is possible to manufacture a foam with a high foaming expansion ratio from pulverized paper, and particularly high-grade waste paper, and this foam can be manufactured inexpensively, at a uniform level of quality, and will be even safer.","Also, with the present invention, in the manufacture of a foam of a plastic resin containing as compositional components pulverized paper, pulverized wood, or another such plant-derived fibrous material and starch, there is a step of adding water to the plant-derived fibrous material and starch and mixing the components to manufacture pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination, and a step of foaming the pellets by mixing them with a plastic resin, and included in between these two steps is a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized wood as a compositional component, the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, or the pulverized wood being of reed or the like, the advantage of which is that a foam of particularly high quality can be manufactured from pulverized wood, and reed in particular.","Also, with the present invention, in the above-mentioned foam manufacturing method, the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %, which allows a foam with a relatively high expansion ratio to be manufactured.","Also, the present invention provides relatively inexpensively a foam manufactured by the manufacturing methods described above, having a high foaming expansion ratio and excellent thermal insulation performance.","BRIEF DESCRIPTION OF THE DRAWINGS FIG.","1 is a concept diagram of the manufacturing method of the present invention; FIG.","2 is a cross sectional micrograph of the present invention; FIG.","3 is a cross sectional micrograph of the present invention; FIG.","4 is a micrograph of the interior of the foam of the present invention; FIG.","5 is a data table of the results of foam manufacture in the present invention; FIG.","6A is a flow state diagram of when the pressure is relatively low in the reservoir in prior art, and FIG.","6B is a flow state diagram of slip cone generation when the pressure is relatively high in prior art; FIG.","7 is a graph of the viscosity of various kinds of starch in prior art; FIG.","8 is a graph of starch viscosity characteristics in prior art; FIG.","9 is a graph of elongation viscosity characteristics in prior art; FIG.","10 is a flow chart of a foam manufacturing method in prior art; FIG.","11 is a simplified diagram of the main parts of a foam manufacturing apparatus in prior art; FIG.","12 is a cut-away oblique view of the raw material extrusion die in the manufacture of a foam board in prior art; FIG.","13 is a manufacturing state diagram of a foam board in prior art; and FIG.","14 is a data table of the results of foam manufacture in prior art.","BEST MODE FOR CARRYING OUT THE INVENTION The present invention will now be described through reference to the drawings.","It was discussed above that it is difficult to mix and melt a plurality of polymer compounds having different viscosities and then extrude and foam this mixture while keeping the fluidity as uniform as possible, if the raw materials are merely mixed.","For the present invention, investigation focused on the mixing of a pulverized paper component and a starch.","The present invention involves creating this mixture by an entirely new idea and manufacturing method that did not exist until now, supplying the mixture thus produced and a plastic resin to a biaxial extruder along with water, and mixing and melting the components to manufacture a foam of uniform quality.","The means for accomplishing this will now be described in specific terms.","Even if the raw material mixing step involves first mixing the pulverized paper with the starch, and then mixing in the propylene or other plastic resin, unless a special method is employed for the first mixing step, uniform fluidity cannot be ensured in the second mixing step that occurs when the components are put into the biaxial extruder, so even though the steps are separate, they will be referred to simply as mixing.","The phrases “mere mixing” and “merely mixing” as used in this Specification are in every case defined in this way.","The starch used in the embodiments is cornstarch, whose agglutination temperature is generally held to be between 80 and 90° C., but since agglutination actually begins as soon as the temperature goes over 60° C., it was found that the agglutination temperature varies with the processing performed by the starch manufacturer.","Further, since the thermal deformation temperature of a plastic resin is 100 to 120° C., the cornstarch agglutinates first and the viscosity increases suddenly, so particularly when the starch is contained in a sufficiently large quantity (weight ratio), it is almost unavoidable that the melt mixing state with the polypropylene resin will be heterogeneous.","This applies not only to polypropylene, but also to polyethylene and other resins, as long as the thermal deformation temperature of the plastic resin is higher than the agglutination temperature of the starch.","It is also obvious that the effect the starch has on the fluid will be completely different if the amount of starch is far too low.","However, the linear polypropylene resin used in the embodiments of the present invention has a low viscous stretching strength and stretching viscosity, which prevents cell growth, and it is said that with a single resin the foaming expansion ratio will be only a few times.","The blend proportion of starch and the function thereof are important if a linear polypropylene resin is to be used to manufacture a foam with a high foaming expansion ratio.","In an effort to solve the problems related to fluidity and viscosity that occur with a mixture having a sufficiently high starch content, the inventors researched the question of whether the agglomeration effect of the starch molecules on the surface of pulverized paper could be effectively utilized in the manufacture of a mixture of pulverized paper and starch.","This idea was the starting point for the present invention, resulting in an extremely useful invention and at the same time yielding a discovery that is significant in terms of understanding the phenomena of agglomeration and colloid formation, which still leave many scientifically unanswered questions.","Starches usually will not dissolve in water.","They disperse in a colloidal state.","The size of molecular spherulites of starch varies with the type of starch, but is said to be from 1 to 200 microns.","In the examples of the present invention, the pulverized paper and starch are stirred and mixed for approximately 60 minutes in a ribbon mixer, after which 1.5 liters of water is added to 3 kg of mixture and kneaded for about 5 minutes in a batch kneader.","This kneaded sol is granulated in a wet granulator.","These granules are called pellets.","After this a dryer is used to perform static drying for approximately 20 hours at 40° C. The pellets are stirred around once or twice during this time to ensure uniform drying.","The highest temperature throughout the kneading and drying steps is 40° C., at which no agglutination of the starch occurs at all.","If the drying temperature is far too high, the starch molecular spheres of the pellets may undergo partial agglutination.","As discussed below, temperature control is very important during the drying process, since it can affect fluidity or even affect the function of the starch as a binder, and hinder the formation of thin cell membranes.","The above-mentioned pulverized paper is high-grade waste paper or discarded postal cards (including postal cards made of materials equivalent to that of postal cards), postage stamps or other such stamps, currency, and so forth, or printing scraps thereof.","The high-grade waste paper referred to in this Specification must meet three conditions: (1) the fiber content must be at least about 8 times the total content of standard waste paper, (2) the material must be safe enough that any hazardous substances such as formaldehyde it contains pass the standards set forth in the test methods of the Japanese “Harmful Substance-Containing Household Article Control Act,” (3) no chlorine or other bleaching agents must be used in the processing of this waste paper.","An example of the above-mentioned pulverized wood is of reed (a perennial grass of the Gramineae family).","Wood of a substance equivalent to the above-mentioned reed is also encompassed.","The particles of pulverized paper or pulverized wood average about 10 to 20 microns in thickness and a few dozen microns in length.","In the case of starch molecular spheres about 20 microns in diameter, the surface area of these particles is sufficient for about 10 to 20 of these spheres to adhere.","The forces at work in agglomeration are said to include the van der Waals force, electrostatic attraction, and the solid crosslinking force, but with molecules adhering to the surface of pulverized paper, the force of mechanical adhesion to microscopic protrusions on the surface cannot be overlooked, and the adsorption strength of starch molecular spheres adhering to the surface is believed to be high.","Judging from the numbers of pulverized paper particles and starch molecular spheres, a number of layers of starch molecular spheres are agglomerated surrounding starch molecular spheres adsorbed to the pulverized paper surface.","Table 1 shows the results of putting specific amounts of two types of pellets that had undergone this agglomeration processing and the starch used as the raw material into a beaker, adding water, and heating and stirring while observing how the state changes with temperature.","TABLE 1 Temperature (° C.) Starch alone Pellets A Pellets B 50 no agglutination 75 agglutination 100 125 no agglutination 150 no agglutination agglutination Pellets A were produced by producing a raw material mixture of equal weights of pulverized paper and starch, kneading this with water in a weight ratio of 2 to 1, and extruding this product at a granulation and extrusion orifice diameter of 1 mm to produce pellets, then drying these for 20 hours at 40° C. Pellets B were produced under exactly the same conditions, except that the granulation and extrusion orifice diameter was 2 mm.","As shown in Table 1, with starch alone, agglutination begins and viscosity rises suddenly at 60° C. or higher, but with a starch that has agglomerated on the pulverized paper surface, no agglutination occurs at this temperature, although there is a colloidal state in which agglomerated starch on the pellet surfaces is partially dispersed.","The only difference between pellets A and B is the granulation orifice diameter.","The agglomeration force and the breakdown thereof are not yet fully understood, but one cannot help but think that the difference is that when the granulation orifice diameter is 1 mm, the extrusion pressure increases and the agglomeration force grows.","For example, there has been research indicating that ceramic firing varies with differences in the agglomeration force balance due to how the material components are agglomerated or with control of the agglomeration structure, and the inventors of the present invention have observed in investigations leading up to the present invention that agglomerated starch molecular spheres are eluted when the pellet drying temperature is varied between 30 and 50° C., that is, that there is a subtle difference in the colloidal state, which makes it extremely interesting that slight differences in conditions can produce differences in agglomeration.","It has been mentioned that many parts of the chemistry of agglomeration are still unclear, but in any case the fact that the agglomerated starch of pellets exhibits behavior completely distinct from that of the agglutination of starch alone is important when considering the fluidity of a mixture containing a plastic resin.","The present invention involves mixing a plastic resin with agglomerated pellets exhibiting the above properties, putting this mixture into a biaxial extruder or the like, and manufacturing a foam.","When a polypropylene resin is used as the plastic resin, since the thermal deformation temperature is only about 20° C. higher with respect to cornstarch, unless there is a sharp increase in agglutination and viscosity of the starch in this temperature region, the fluidity of the polypropylene is the main factor in determining the fluidity of the entire mixture.","This the problem to be solved, namely, the above-mentioned governing viscosity and fluidity of the system.","When the agglomerated pellets and the plastic resin are put into the biaxial extruder and while the mixture is proceeding to the discharge orifice, it is believed that the pellets begin to melt while containing relatively small amounts of starch colloid dispersed from the pellets, resulting in a state of being surrounded by polypropylene.","It is surmised that agglutination of the starch dispersed from the pellets occurs in the high-temperature cylinder at the end, and finally the starch molecular spheres near the pulverized paper surface undergo agglutination.","This incremental agglutination theory would seem to be the correct view, judging from the shape of the foam as discussed below.","FIG.","2 is a cross sectional micrograph of a foam produced using a linear polypropylene (one with no branches), pellets of cornstarch, and pulverized paper produced as above.","The compositional ratios (in weight percent) were 28%, 24%, and 48%, respectively.","It can be seen right away that the foaming expansion ratio is very high despite the use of a linear polypropylene, with which it is held that a foam with a high foaming expansion ratio cannot be obtained because the stretch viscosity is low.","Naturally, the higher the foaming expansion ratio, the thinner the cell membranes, and the membrane thickness is just a few microns.","With a linear polypropylene alone, it is said that the foaming expansion ratio will only be about 2 to 3 times, and cross sectional views provided by manufacturers reveal the membrane thickness to be several dozen microns.","Taking a look at why, in contrast, a foam with such thin membranes is formed from a polypropylene with the same low stretch viscosity when mixed with pulverized paper and cornstarch as in the present invention is also important in terms of understanding the incremental agglutination of starch and how the fluidity of the system is governed.","When a foam is manufactured from a single plastic resin, small melting-resistant particles commonly called frothers are added, mixed, and stirred, and it is said that a foam is produced through the growth of microscopic bubbles called nuclear cells, which are generated as a result of the different phases.","The growth of these cells is said to be determined exclusively by the stretch viscosity and surface tension of the resin.","However, it would seem to be impossible to explain the formation of the thin membranes in the present invention by this widely-held theory alone.","We probably have no choice but to conclude that the situation is somewhat different for the foam formation mechanism of a mixture that contains relatively large amounts of polymer materials of different viscosities and that contains pulverized paper, whose properties and shape remain basically unchanged at the temperatures in all of the steps.","Nuclear cells are generated when the flow of a fluid with different viscosities becomes turbid.","It is believed that nuclear cells are readily generated in many locations during the foam manufacturing process of the present invention, in which a fibrous material is admixed, and that the number of these cells is extremely large.","When these countless nuclear cells are discharged under reduced pressure from a biaxial extruder, they grow all at once.","As said in the case of a single resin comprising linear polypropylene, if the stretch viscosity of the resin is low, most of these nuclear cells will not grow completely, resulting in cell clusters, and as a result the foaming ratio is lower and the cell membranes are thicker.","However, with the foam of the present invention containing a starch, it seems that a major role is played by the starch, in which changes in the agglutination state arise from condensation, and the pulverized paper fibrous material.","The amount of starch component that mixes with the molten polypropylene is relatively small compared to the amount of raw materials added, and it is reasonable to conclude that most of the starch stops around the pulverized paper.","There should be a local increase in viscosity as a result of the agglutination of starch eluted while the polypropylene is molten, but since the amount is not very large, the mixture as an entire system seems to exhibit relatively stable fluidity.","As discussed previously regarding the colloid formation that accompanies an increase in temperature, the agglomerated starch adhering around the pulverized paper is not dispersed and eluted all at once, but instead gradually disperses from the agglomeration surface.","Near the last stage of the biaxial extruder, that is, the discharge orifice, all of the starch is believed to be agglutinated, but pellets are present in dispersed form in the completely melted polypropylene resin fluid, and the concentration distribution of amylose in the agglutinated starch is believed to have a gradient going outward from around the pulverized paper, that is, toward the main polypropylene fluid.","It is surmised that, despite their low stretch viscosity, the polypropylene membranes that have joined together with the partially agglutinated starch are given a cushioning effect by the elasticity and stretchability of the agglutinated starch surrounding the pulverized paper, and at the same time a balance is achieved in the extremely thin membrane state through the pressure of the adjacent cells that are similarly undergoing rapid growth.","The importance of temperature management in the pellet drying process has already been discussed, but if the pellets end up being partially agglutinated in the drying process, this will affect the cushioning effect resulting from the elasticity and stretchability of the starch.","FIG.","3 is a photograph of a foam that has been cut diagonally.","A relatively thick film can be seen surrounding the pulverized paper fibrous material, and we see that there is a continuous reduction in thickness.","FIG.","4 is a photograph at 10,000-times magnification of the interior of a foam, taken with a scanning electron microscope.","A film in which tiny holes a few dozen to a few hundred nanometers in size can be seen is formed between two objects that seem to be fibrous material about 1 micron in size, and that look thick in the photograph.","This film is not a cell membrane, but these microscopic observation results seem to bear out the foam membrane formation mechanism of the present invention that has been inferred up to now.","As expected, the primary means for solving the problem of the present invention does indeed solve the problem of too much variance in the raw material mixture inside the biaxial extruder by making use of pellets in which the agglomeration force of starch and pulverized paper is put to suitable use, but at the same time the function of starch as a binder, which up to now had only been described in vague terms, becomes more clear, and this is a groundbreaking discovery that is the key to forming extremely thin cell membranes.","As discussed above, an embodiment of the present invention relates to the flow stability of a mixed molten fluid, which is a problem in the manufacture of a foam of uniform quality using a plurality of polymer compounds with different melting temperatures or viscosities.","The components that make up the foam consist of a pulverized fibrous material that will not melt at the temperatures encountered during manufacture, such as wood or paper whose main component is cellulose, lignin, or the like; starch, which is necessary as a binder for the fibrous material and the plastic resin; a plastic resin having a thermal deformation temperature higher than the agglutination temperature of the starch; and water or the like as a foaming agent.","Furthermore, the present invention is the result of a finding that confirms the correctness of the theory based on typical experiments and the theory developed from the results of a number of experiments relating to the function of agglomerates in mixed fluids in which a plastic resin is melted, and that agglomerations of starch molecular spheres are produced, the core of which is made up of starch, which undergoes a major thermal reaction (a phenomenon called agglutination) at the lowest temperature of any of the constituent components other than water, and a pulverized fibrous material of paper, wood, or the like that has substantially no temperature dependence in terms of the fluidity of the mixture, and the reactivity of the agglomerated starch with respect to temperature is different from that of the raw material starch.","Therefore, in the foam constituent raw materials mentioned in the test and embodiments cited in this Specification, except for the starch, the fibrous material is not limited to pulverized paper as long as the thermal change defined in the present invention does not occur within the temperature range encountered in the foam manufacturing process.","Similarly, a linear polypropylene resin, which is an inexpensive and readily available raw material, is used in the present invention as a typical example of a safe plastic resin that does not give off dioxins or other such harmful substances when disposed of and incinerated, but the plastic resin is not limited to a polypropylene resin as long as it has fluidity and stretchability suited to the manufacture of a foam and has a thermal deformation temperature higher than the agglutination temperature of the starch.","Also, water is used as the medium in the present invention to manufacture an agglomerate of starch by adsorbing starch molecular spheres to the plant-derived fibrous material that serves as the core in the present invention, but there are other methods for producing an agglomerate as well, such as causing the substance particles to collide with each other without the use of water, so the agglomerated starch of the present invention may be obtained by any method as long as it yields an agglomerate that exhibits the behavior described in detail in the present invention when melted along with other polymer compounds.","For the same reason, regarding the foaming method, steam foaming in which water serves as the foaming agent is performed in the present invention, and to this end the tests and examples described herein make use of a biaxial extruder, but the present invention is not limited to this.","Furthermore, a foam of uniform quality can be manufactured in the present invention by having the plant-derived fibrous material (pulverized paper or pulverized wood) component account for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.","EXAMPLES Example 1 Example 1 relates to a cushioning material obtained by cutting a string of foam into pieces.","This cushioning material is used to protect objects during shipping.","A specific manufacturing method and steps will be described through reference to the flow chart of FIG.","1.Pellet Production Step Using postal card waste paper as the plant-derived fibrous material, first the paper was coarsely pulverized in a mixer/cutter type of pulverizer, and then it was finely pulverized in a ball mill type of pulverizer.","This product was sifted through an industrial sieve to obtain a waste paper raw material of the required size.","Next, this waste paper raw material was mixed in an amount of 40% (weight ratio) with cornstarch in a weight ratio of 60% for 60 minutes in a ribbon mixer.","3 kg of this mixture was combined with 1.5 liters of water and kneaded for 5 minutes in a batch kneader.","Pellets were produced in a wet granulator and allowed to dry for 12 hours in a dryer set at 50° C., after which the pellets were stirred around and dried for another 8 hours to ensure even drying.","Biaxial Extrusion Step The dry pellets were mixed with a linear polypropylene resin (as the plastic resin component), a suitable amount of water was supplied, and the components were kneaded well.","This kneaded raw material was fed into a biaxial extruder A and foamed.","The biaxial extruder shown in FIG.","11 comprises a raw material inlet 1, a water inlet 2, cylinders C1 to C5, heaters 3 attached to the various cylinders, a raw material extrusion die (spinneret) 4, and an extrusion opening 5.In manufacturing tests, the settings of the cylinder and extrusion die temperatures was suitably varied.","The test was conducted with the water supply varied between 10 and 30 liters per hour.","These results are given in FIG.","5.The results from FIG.","5 are also given in Table 2 in comparison with the results from FIG.","14 (prior art).","TABLE 2 Comparison table Simple Subject Compared category Pellets1 mixture2 Unit Average length 5.602 4.874 length (cm) Size of range 2.5 6.6 (cm) Maximum length 6.6 8.5 (cm) Minimum length 4.1 1.9 (cm) Standard 0.634 1.923 deviation Unit Average length 2.074 1.886 length (cm) Size of range 0.4 1.4 (cm) Maximum length 2.3 2.4 (cm) Minimum length 1.9 1.0 (cm) Standard 0.110 0.328 deviation [1present invention, 2prior art] It can be seen that there is far less variance in the foam manufactured by the technique of the present invention than in the foam manufactured by prior art.","Also, 29 out of 50 samples were within ±10% of the average length with the present invention, whereas the number was only 4 with the prior art.","What is even more noteworthy is the difference in regard to diameter.","In Table 2, the difference in the variance in foaming extent in FIGS.","5 and 14 is expressed directly as the size of the range of variance in diameter.","The uniformity of the extent of foaming and the small variance in diameter and length achieved with the technique of the present invention make it possible to manufacture a foam board, which was impossible in terms of quality with prior art.","Example 2 Next we will describe the manufacture of a foam board by proceeding through the same steps as in Example 1, but with the extrusion die (spinneret) of the biaxial extruder changed.","The extrusion die (spinneret) used in the prior art and Example 1 was intended for manufacturing a foam string, being equipped with two discharge orifices of 2 mm a suitable distance apart, but as shown in FIGS.","12 and 13, the structure of an extrusion die for manufacturing a foam board is provided with numerous 2-mm discharge orifices and has a box-like area to the rear thereof.","A foam board can be manufactured by passing the material through this die.","Foam boards can be applied to the cardboard insides of shipping containers that need to offer thermal insulation.","After use, the insulating foam boards can be removed, and can be treated as waste material that will not give off dioxins or other harmful substances when burned.","This makes such boards very useful as a thermal insulation material.","Since the size of the cardboard container is set, if the dimensional precision of the foam board is off by too much, the board will be too long to fit into the container, or it may be too short, so that it does not stay pressed against the cardboard container and falls over, or there may be gaps in the corners, which compromises the thermal insulation performance of the container, so automatic dimensional precision is important from the standpoint of industrial production.","In view of this, we wanted to test how much variance in cut length there was in the foam board of the present invention with respect to the cut length target, and to confirm the foam quality, and therefore examined the thermal insulation of a container in which foam boards were actually installed inside a cardboard container.","Table 3 shows the cutting variance test results.","In approximately 2 hours of manufacture, a foam board with an overall length of as large as 110 meters was manufactured, with the cutting target set at 1000 mm.","Therefore, 110 foam boards cut to 1000 mm were produced.","In this test, 20 random samples were collected and measured for length.","TABLE 3 Variation in length of foam boards Sample No.","Length (cm) 1 100.5 2 100.1 3 100.3 4 99.9 5 100 6 100 7 100.6 8 100.3 9 99.7 10 100 11 100.1 12 100.8 13 100.2 14 99.5 15 99.8 16 100 17 99.8 18 100.3 19 100.3 20 100.1 Average 100.115 Standard deviation 0.311659 As a result, the average length was 100.115 cm, the maximum length was 100.6 cm, and the minimum length was 99.5 cm, giving a standard deviation of 0.312, which means the results were extremely stable.","Thus, a foam board could be molded, which could not be obtained with prior art.","What is even more interesting is that while the average length of one of the foam strings produced by the technique of the present invention shown in Table 2 of Example 1 was 5.6 cm and the standard deviation was 0.634, but the average length of the foam boards was 100.1 cm, with a standard deviation of only 0.312.With the foam strings, the time it took for a foam of 5.6 cm to be produced was calculated to be a mere 0.028 second.","By contrast, with the boards, the foam discharge was slowed by the difference in pressure arising from the shape of the extrusion die (spinneret).","The time it took to produce a foam board with a length of 1 meter was approximately 1.2 seconds.","In other words, this indicates that inside the biaxial extruder the fluidity fluctuates from instant to instant (0.028 second), but when viewed at a time of 1.2 seconds the fluidity of the system is extremely stable.","This proves quite conclusively the correctness of the theory about the fluidity of a composite polymer compound discussed in detail in the prior art section of this Specification, and the correctness of the approach of the method for solving the targeted problem in the present invention.","Next, we examined the insulation performance of a cardboard container in which these foam boards were used.","For this evaluation we made a comparison with a foamed polystyrene container, which is the most common type of insulating container in use today.","The containers used here were as follows.","1.Cardboard Container in Which Foam Boards Were Used This was a cardboard container lined on the insides with insulating foam boards (panels) produced according to the present invention using a waste paper component.","Container outside dimensions: 290 mm wide×205 mm deep×222 mm high Insulation thickness: 20 mm of foam board Container inside dimensions: 245×160×170 mm 2.Commercially Available Foamed Polystyrene Container Container outside dimensions: 293 mm wide×215 mm deep×220 mm high Insulation thickness: 25 mm Container inside dimensions: 245×165×175 mm Measurement Principles and Method: 1) The container is placed under constant ambient temperature conditions, and then heated with a heater/fan unit inside the container.","2) 2.","[sic] Power is applied to the heater unit until there is a sufficient temperature differential between the container ambient temperature and the average temperature at three points inside the container.","3) Enough time is allowed to pass for the temperature inside the container to become constant (referred to as a steady state).","This means that the temperature inside the container and the ambient temperature (constant) reach a state of equilibrium via the insulated container.","4) The adiabatic coefficient (or heat leakage coefficient) of the container per 1° C. temperature differential is calculated from the power (W) applied to the heater unit and the difference between the ambient temperature and the temperature inside the container in a steady state.","This is expressed as W/° C. Measurement was conducted by the following method.","A temperature measurement jig was placed in the approximate center of the container, the internal[5] height of the container was divided into four equal parts, and the temperature at three height points thereof was measured with a copper constantan thermocouple.","Variance in the temperature inside the container was reduced by the fan of the heater unit.","The ambient temperature was set by the controller of a climatron or a thermostatic tank, but the reliability of the temperature measurement was enhanced by measuring the temperature near the container with a constantan thermocouple to confirm how well it matched the measured temperature inside the container.","The heater unit consisted of a heater mounted on a heat sink, and a fan used to diffuse the heat radiated from the heat sink.","The heater and the fan were driven by separate DC power supplies, the voltage and current of each were measured, the total applied power was calculated, and this was termed the amount of heat generated inside the container.","Thermal insulation performance was measured at two locations externally, and are termed measurement test sites A and B.","These results are given in Table 4.In all of these measurements, the cardboard container to which the foam boards had been applied (also called a “Super-Chilled Echo U Container”) exhibited insulating performance at least several percentage points higher than that of the foamed polystyrene container.","The thickness of the insulating material applied to the cardboard container to which the foam boards had been applied was 20 mm, whereas the thickness of the foamed polystyrene container was 25 mm, so if we consider materials of equal thickness, the results for the insulating material of the cardboard container to which the foam boards had been applied suggest that this foam has thermal insulation properties at least 25% better than those of standard foamed polystyrene.","TABLE 4 Table of measurement results and calculated adiabatic coefficients Measurement test location Measurement site A Measurement site B Test apparatus Thermostatic tank Climatron Measured container* foamed foamed board polystyrene board polystyrene Ambient temperature 19.9° C. 19.8° C. 15.1° C. 15.6° C. Average temperature 51.0° C. 48.2° C. 49.3° C. 47.2° C. inside container Temperature 31.1° C. 28.4° C. 34.2° C. 31.6° C. differential Heater input power 9.53 W 9.60 W 9.65 W 9.67 W Fan input power 0.148 W 0.148 W 0.161 W 0.163 W Total heater unit 9.68 W 9.75 W 9.81 W 9.83 W input power Container adiabatic 0.311 W/° C. 0.343 W/° C. 0.278 W/° C. 0.311 W/° C. coefficient *Measured containers: “Foamed board” is short for a cardboard container lined with foamed boards, while “polystyrene” is short for a foamed polystyrene container used as a control.","As seen in Table 4, the adiabatic coefficient of the cardboard container to which the foam boards had been applied was 0.311 W/° C. at measurement site A and 0.287 W/° C. at B, while the adiabatic coefficients of the control foamed polystyrene container were 0.343 W/° C. and 0.311 W/° C., respectively.","The result is that the insulating performance of the cardboard container to which the foam boards had been applied was excellent (10.3% at measurement site A and 8.4% at B).","However, the adiabatic coefficient varies by about 10% between the two measurement sites.","The main reason for this seems to be the difference in the amount of air circulating inside the thermostatic apparatus keeping the ambient temperature of the container constant.","In other words, at measurement site B the measurement was made in a relatively large climatron in which people can enter and work, whereas measurement site A was a much smaller thermostatic tank.","The amount of air moving in a thermostatic tank generally tends to be large, and if the amount of air moving around an insulated box is larger, this is equivalent to the insulated container having a larger surface area, so the analysis was that the apparent insulation performance generally decreases.","Therefore, a relative comparison under equal conditions is important in performing an evaluation such as this.","This measurement and evaluation involved measuring the insulation performance of the same container at difference measurement test locations, and since the results for both tended to be the same, with an extremely small difference therebetween, and with the above-mentioned effect of the amount of air movement factored in, it was concluded that the data were a good match.","In Example 2 above we discussed the manufacture of a foam board, but a foam sheet can also be manufactured by providing a roll molding machine comprising an upper roll and a lower roll to the rear of this manufacturing apparatus, and squeezing the above-mentioned foam board in its thickness direction by passing it through this roll molding machine.","Industrial Applicability With the present invention, when a foam is manufactured using a starch as one polymer compound to be melted and mixing it with another polymer compound having a different melt viscosity, uniformity is ensured in the fluidity of the molten mixture, which is an essential requirement for stabilizing foam quality, and the resulting foam will have thin cell membranes and excellent thermal insulation performance, allowing a high-quality foam to be manufactured stably on an industrial scale.","Also, this foam will be favorable in terms of protecting the global environment."]],"string":"[\n [\n \"Process for producing foarmed articles and the foamed article\",\n \"It is an object of the present invention to provide a method for manufacturing a high-quality foam that can be produced stably on an industrial scale and that affords excellent thermal insulation as a foam with thin cell membranes, which method ensures uniform molten mixture fluidity, which is an essential condition for achieving consistent foam quality, in the manufacture of a foam by mixing starch as one molten polymer compound with another polymer compound having a different melt viscosity, and to provide a foam manufactured by this method.\",\n \"Water is added to and mixed with a plant-derived fibrous material and starch to manufacture a foam of a plastic resin containing as compositional components starch and a plant-derived fibrous material such as pulverized paper, pulverized wood, or the like.\",\n \"A foam is manufactured by a method comprising a step of producing pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination, and the step of foaming the pellets by mixing them with a plastic resin, and included in between these two steps is a step of drying the pellets at a temperature not higher than the starch agglutination temperature.\"\n ],\n [\n \"1.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing the pellets with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature.\",\n \"2.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower.\",\n \"3.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component.\",\n \"4.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component.\",\n \"5.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the pulverized paper is made from high-grade waste paper, or from printing scraps or waste of postal cards, stamps such as postage stamps, currency, and so forth.\",\n \"6.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the pulverized paper is made from high-grade waste paper, or from printing scraps or waste of postal cards, stamps such as postage stamps, currency, and so forth.\",\n \"7.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized wood as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component.\",\n \"8.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, the plant-derived fibrous material comprising pulverized wood as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component.\",\n \"9.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized wood as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the pulverized wood is of reed or the like.\",\n \"10.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, the plant-derived fibrous material comprising pulverized wood as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the pulverized wood is of reed or the like.\",\n \"11.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing the pellets with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.\",\n \"12.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.\",\n \"13.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.\",\n \"14.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.\",\n \"15.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the pulverized paper is made from high-grade waste paper, or from printing scraps or waste of postal cards, stamps such as postage stamps, currency, and so forth, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.\",\n \"16.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the pulverized paper is made from high-grade waste paper, or from printing scraps or waste of postal cards, stamps such as postage stamps, currency, and so forth, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.\",\n \"17.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized wood as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.\",\n \"18.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, the plant-derived fibrous material comprising pulverized wood as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.\",\n \"19.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized wood as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the pulverized wood is of reed or the like, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.\",\n \"20.A foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, the plant-derived fibrous material comprising pulverized wood as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the pulverized wood is of reed or the like, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.\",\n \"21.A foam, manufactured by the foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing the pellets with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.\",\n \"22.A foam, manufactured by the foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.\",\n \"23.A foam, manufactured by the foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.\",\n \"24.A foam, manufactured by the foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.\",\n \"25.A foam, manufactured by the foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the pulverized paper is made from high-grade waste paper, or from printing scraps or waste of postal cards, stamps such as postage stamps, currency, and so forth, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.\",\n \"26.A foam, manufactured by the foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the pulverized paper is made from high-grade waste paper, or from printing scraps or waste of postal cards, stamps such as postage stamps, currency, and so forth, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.\",\n \"27.A foam, manufactured by the foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized wood as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.\",\n \"28.A foam, manufactured by the foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, the plant-derived fibrous material comprising pulverized wood as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.\",\n \"29.A foam, manufactured by the foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized wood as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the pulverized wood is of reed or the like, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.\",\n \"30.A foam, manufactured by the foam manufacturing method, for manufacturing a foam of a plastic resin containing, as compositional components, starch and a plant-derived fibrous material such as pulverized paper, pulverized wood or the like, comprising the steps of: adding water to the plant-derived fibrous material and starch and mixing the components to produce pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination; and foaming said pellets by mixing them with a plastic resin, and the method further comprising, between these two steps, a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the pellets being dried for a relatively long time at a low temperature of 50° C. or lower, the plant-derived fibrous material comprising pulverized wood as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, wherein the pulverized wood is of reed or the like, wherein the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.\"\n ],\n [\n \"<SOH> BACKGROUND ART <EOH>There have been countless attempts at manufacturing a foam by mixing starch with another plant-derived material or with a plastic resin that would pose no danger of emitting dioxins if incinerated after use, for example, and at utilizing such foams as thermal insulation materials or as cushioning materials or other such shipping and packaging materials, and a number of methods for manufacturing foams have been disclosed.\",\n \"Nevertheless, based on an examination of this disclosed information, it appears that no known technique has been established relating to the problem the present invention is intended to solve.\",\n \"To clarify the problem the present invention is intended to solve, general concepts, beliefs, related examples, and so forth were examined in the technological field to which the present invention is related.\",\n \"It is commonly held that when steam or the like is used as a foaming gas, the material is discharged under pressure from an extrusion molding machine, and reduced-pressure foaming is performed, for instance, a polymer raw material that becomes fluid when the temperature is raised will flow as shown in FIG.\",\n \"6A , with symmetrical flow lines around the center line in the drawing, when the pressure is relatively low, and as the pressure is gradually raised, slip is observed between the material and the walls.\",\n \"At even higher pressures the flow lines begin to be disarrayed, the material is extruded in a spiral shape, and this results in a lack of uniformity in the material.\",\n \"What happens here is called stick and flow, in which the flow of the material comes to a halt (sticking), after which the material starts sliding again (slipping).\",\n \"When this occurs, complete and intermittent slipping over the entire surface of the slip cone shown in FIG.\",\n \"6B has been reported for high density polyethylene and other such polymer materials, and a similar phenomenon is seen not just with polymer compounds but also in the extrusion of bismuth-tellurium and other such metal materials that are thermoelectric semiconductors, for example.\",\n \"This is manifested as cracks in the extruded semiconductor.\",\n \"Various apparatus improvements have been attempted from a mechanical aspect in an effort to eliminate this problem, but the fact is that the fluid properties of polymer compounds are still not fully understood by researchers.\",\n \"The materials that are the subject of the present invention are polymer compounds, and it is necessary to understand this phenomenon.\",\n \"Since the present invention involves a plurality of polymer compounds, however, it is uncertain whether the phenomenon witnessed with a single compound such as polyethylene would be the same, and the problem is even more complicated because we are dealing with a mixture in which the temperature dependency of viscosity and the thermal deformation temperature characteristics vary with the different raw material polymer compounds, which probably makes it even more difficult to understand the situation in terms of fluid dynamics or tribology.\",\n \"The present invention relates to a field of technology in which such scientific understanding is not necessarily perfect, but at the very least, of course, we must know something about the material characteristics of the raw material polymer compounds being used.\",\n \"Let us now give a brief summary of the characteristics of a polypropylene resin, as an example of a plastic resin, and starch, which are believed to be the main contributors to fluidity out of the polymer compound materials used in the present invention.\",\n \"Starch is usually in the form of macromolecular spherulites of a mixture of two homologues with different structures, amylose and amylopectin.\",\n \"The size thereof can vary greatly with the type of starch, but the diameter is said to range from 1 to 20 microns, and the molecular weight up to a few thousand.\",\n \"Amylose is a linear polymer to which D-glucose units are connected by α-1,4 linkages, whereas amylopectin is a branched polymer that includes α-1,6 linkages in addition to α-1,4 linkages, with branching occurring at these sites and the branches connected again with α-1,4 linkages.\",\n \"When starch granules are suspended in water and heated to a certain temperature or higher, the starch granules undergo irreversible swelling and amylose is eluted.\",\n \"This process is called agglutination, and this temperature is referred to as the agglutination temperature.\",\n \"FIG.\",\n \"7 shows published heating and viscosity curves for various kinds of starch (amylograms).\",\n \"The agglutination temperature ranges from 60 to 90° C., and cornstarch is what is cited in the embodiments and so forth of the present invention.\",\n \"FIG.\",\n \"8 also gives published information, and is a comparison of the viscosity behavior of ordinary wheat resulting from agglutination, by variety of glutinous wheat (which has a relatively high amylopectin content), versus that of cornstarch.\",\n \"For this comparison, the temperature of each starch was raised from 34° C. to 94° C. at a rate of 5° C. per minute, after which the starch was cooled at the same rate of 5° C. per minute and its viscosity was measured.\",\n \"Of these varieties, Norin No.\",\n \"61 had the same amylose content of 31.8% as ordinary wheat, and its agglutination temperature was close to that of wheat as shown in FIG.\",\n \"8 .\",\n \"The waxy cornstarch used as a control also has a relatively low agglutination temperature, similar to what is shown in FIG.\",\n \"7 .\",\n \"The purpose of providing FIGS.\",\n \"7 and 8 here is to show the agglutination temperature of the starch that is one of the polymer compound materials used in the present invention in relation to the fluidity of these materials, and demonstrate that there is a rapid increase in viscosity as soon as the agglutination temperature is attained.\",\n \"The polypropylene resin will now be discussed.\",\n \"Polypropylene is a crystalline thermoplastic resin made by the polymerization of propylene (C 3 H 6 ), and is hard and tough, with good resistance to moisture, oils, and solvents.\",\n \"Its heat resistance temperature is said to be 170° C. or lower.\",\n \"Also, there is almost no possibility that a linear polymer compound will undergo cyclization during combustion, and these resins, along with polyethylene and the like, are called polyolefin resins and said to be resins that do not produce dioxins or other harmful substances.\",\n \"According to disclosed reference materials from polypropylene manufacturers and so forth, the thermal deformation temperature of polypropylene varies somewhat with the structure of the compound, but is said to be roughly 100 to 120° C. FIG.\",\n \"9 shows an example of the elongation viscosity of the linear (without branches) polypropylene resin used in the examples and so forth of the present invention, and it can be seen that the elongation viscosity at 180° C. quickly disappears in just a few seconds.\",\n \"The reason a foam of a high expansion ratio cannot be produced from a linear polypropylene resin is said to be that the elongation viscosity drops off so quickly and the melt tension is low, and most polypropylene manufacturers produce polypropylene resins having a branched structure by subjecting a linear polypropylene to a special process, and then make a foam with a relatively high expansion ratio from this polypropylene resin with improved elongation viscosity and melt tension.\",\n \"Higher cost is at the present time inevitable with a branched polypropylene because a linear polypropylene must be subjected to a post-treatment step such as electron beam irradiation.\",\n \"When an attempt is made to produce a foam by mixing and melting a starch and a plastic resin such as polypropylene, the difference between the temperature dependence of the viscosity behavior and the melt temperature of the polymer compounds results in complex fluidity of the mixed materials in the mixing, melting, and extrusion steps during manufacture, and it is not hard to imagine the effect that this has on discharge uniformity.\",\n \"Japanese Laid-Open Patent Application H9-111029 discusses a method for manufacturing a loose polypropylene foam by mixing a polypropylene resin and a starch-based additive into a plant-derived foaming agent such as nonfat powdered milk or bean-curd lees, putting this raw material mixture into an extruder comprising a high-temperature heated cylinder, and feeding in water.\",\n \"This publication also discusses a method in which a plastic component, plant-derived foaming agent, and starch-based additive are simply mixed.\",\n \"The inventors of the present invention conducted manufacturing tests in which they used an actual foam manufacturing apparatus similar to that in the disclosed information, used as their raw materials a plastic resin, starch, and pulverized waste paper (as the plant-derived fibrous material), and varied the amount of feed water (the foaming agent), the mixing time and temperature in the manufacturing process, the conditions of the material discharge orifice, and so forth.\",\n \"As a result, with a simple mixture of raw materials as in the disclosed information, perhaps because variance in fluidity could not be suppressed, there was considerably fluctuation in the discharge amount per hour, and there was also variance in the foaming state, so it was found that a foam of stable quality could not be manufactured.\",\n \"The specific manufacturing method and steps here are illustrated in FIG.\",\n \"10 .\",\n \"Pulverized paper (such as waste paper) was used as the plant-derived fibrous material, but first the material was finely pulverized in a mixer/cutter type of pulverizer and then in a ball mill type of pulverizer, this pulverized material was sifted through an industrial sieve to obtain a waste paper powder with particles averaging 20 to 30 microns in size, and this powder was used as the waste paper raw material.\",\n \"This waste paper raw material was then mixed with a polypropylene resin to obtain a raw material mixture, to which cornstarch was further added and mixed, and these components were put in a mixing machine and mixed.\",\n \"The sequence of this mixing was also varied so that the waste paper raw material was first mixed with the cornstarch to obtain a raw material mixture, to which the polypropylene resin was added and mixed in a mixing machine.\",\n \"The mixing was also performed at various blend ratios, including those given in the disclosed information.\",\n \"This kneaded raw material was put into,a biaxial extruder and foamed.\",\n \"Specifically, the biaxial extruder shown in FIG.\",\n \"11 comprises a raw material inlet 1 , a water inlet 2 , cylinders C 1 to C 5 , heaters 3 attached to the various cylinders, a raw material extrusion die (spinneret) 4 , and an extrusion opening 5 .\",\n \"In manufacturing tests, the settings of the cylinder and extrusion die temperatures was suitably varied.\",\n \"The test was conducted with the water supply varied between 10 and 30 liters per hour.\",\n \"To evaluate the foam state and foam variance, a string-like piece of foam extruded and foamed from an extrusion die with a diameter of 2 mm was cut up by an automatic cutter set to a specific rotating speed, 50 pieces were randomly sampled from among the total of 4200 pieces of cut foam produced in one minute, whose combined length was 120 m, the length and diameter of the cut foam pieces were measured, and the extent of foaming was visually evaluated on a ten-point scale.\",\n \"The mixing ratios of the materials for which the results are shown in FIG.\",\n \"14 were 25% (weight ratio) pulverized waste paper, 30% starch, and 45% polypropylene resin.\",\n \"As indicated by these results, the foam manufactured by the mixing process described above exhibited considerable variance in all measurement categories.\",\n \"The length of cushioning material cut at regular time intervals varied from a minimum of 1.9 cm to a maximum of 8.5 cm.\",\n \"The most serious problem was variance in the extent of foaming.\",\n \"The ten-point scale assumes 10 to be the ideal foaming state, but because there was variance in the foaming, the cushioning material varied from thin to thick.\",\n \"This indicates nothing other than a change over time in the fluidity of the raw material mixture during mixing and melting inside the cylinders.\",\n \"This is similar to the stick and flow phenomenon described above, but it is unclear whether it occurred because a slip cone was formed, or just because of local variance in the fluidity of the molten mixture.\",\n \"Admixed in this molten mixture was pulverized waste paper, whose characteristics remained almost unchanged over the temperature range in this process.\",\n \"This pulverized paper consists of a fibrous material with a microscopic length of anywhere from a few microns to about 100 microns, which is stirred and mixed in the biaxial extruder, and it seems that nuclear cells that will serve as the nuclei for cells are produced here and there in an agitated fluid in which polymer compounds of different viscosity are admixed.\",\n \"In this situation it is doubtful whether a nice slip cone is being formed as in the above-mentioned reports on polyethylene resins.\",\n \"On the contrary, it seems likely that this nucleus formation is occurring as a result of local variance in the fluidity of the molten mixture.\",\n \"At this point we will briefly discuss the correlation between the viscosity of the above-mentioned starch and polypropylene resin and the fluidity of the mixture of polymer compounds, since such a discussion will probably further clarify the points involved in the problems which the present invention is intended to solve and the means used to solve the problems.\",\n \"A capillary type of viscosity measurement apparatus is the most commonly used in the measurement of the viscosity of polymers.\",\n \"A sample is extruded from a capillary tube by applying pressure, and the viscosity is measured by measuring the difference in the applied pressure at a constant flow rate per unit of time.\",\n \"We will let L be the length of the capillary tube, and r be its radius.\",\n \"The change over time in the amount of sample flowing out of the capillary tube is found for when pressure is applied such that the sample contained in the reservoir flows at a specific rate, and this is termed Q (cm 3 /sec).\",\n \"A relationship of in-line-formulae description=\\\"In-line Formulae\\\" end=\\\"lead\\\"?\",\n \"γ=4 Q/πr 3 in-line-formulae description=\\\"In-line Formulae\\\" end=\\\"tail\\\"?\",\n \"exists between the shear rate γ (sec −1 ) and the sample flowing through the capillary tube, and the shear stress τ (N/cm 2 ) on the inner walls of the capillary tube is expressed as τ=rP/2L.\",\n \"The viscosity μ (N·sec/cm 2 ) is defined from the shear rate y and the shear stress τ as: in-line-formulae description=\\\"In-line Formulae\\\" end=\\\"lead\\\"?\",\n \"μ=τ/γ.\",\n \"in-line-formulae description=\\\"In-line Formulae\\\" end=\\\"tail\\\"?\",\n \"If we assume that a starch and a polypropylene resin are mixed and the thermal characteristics of each are independently manifested, the intrinsic viscosity should be exhibited in the starch at the agglutination temperature and in the polypropylene resin at the thermal deformation temperature, with the viscosity subsequently varying according to temperature changes.\",\n \"For instance, if the agglutination temperature of the starch is reached before the thermal deformation temperature is reached, the viscosity of the starch increases suddenly, and the entire system is governed by the viscosity of the starch.\",\n \"If the thermal deformation temperature of the polypropylene resin is never reached, the polypropylene might have some kind of effect as an admixture, but the viscosity μ of the system will be relatively stable.\",\n \"In other words, this is a state in which the fluidity of the system is governed by the viscosity of the starch, even though the system is a mixture.\",\n \"However, this never happens for the purposes of manufacturing a foam.\",\n \"Still, if the temperature continued to rise, it would eventually reach the thermal deformation temperature of the polypropylene resin, resulting in a fluid having both the viscosity of polypropylene and the viscosity of the starch, which varies with time and temperature.\",\n \"It is very difficult to know what the mixing state is in a mixed fluid such as this.\",\n \"Measurement was made at a certain specific temperature with the above-mentioned viscosity measurement method, but since the mixing is not completely uniform in a mixing state of different viscosities, the local shear rate γ should vary.\",\n \"Furthermore, the shear stress τ on the inner walls of the capillary tube also varies according to the local mixing state.\",\n \"In other words, the viscosity μ of a mixture of polymer compounds changes with how uniform the mixing is, and if we look at the viscosity definition of the above formula, we can see that there is extreme instability in which both the denominator and the numerator vary.\",\n \"Put another way, there is no viscosity that governs the mixture as a system, and instead the system is in a state of constant change.\",\n \"Japanese Laid-Open Patent Application 2000-143869, which is another source of disclosed information, discloses a technique of manufacturing a foam by pulverizing a plastic laminate and mixing with polypropylene, cornstarch, and calcium carbonate.\",\n \"With this technique, the foaming expansion ratio is about 10 times with just the resin that is admixed in the laminate paper, and the admixture of another plastic is proposed in order to increase the foaming expansion ratio.\",\n \"Close examination of this technique reveals that, as seen in the embodiments and examples of the invention, a foamable composition is made up of plastic from pulverized laminate paper and added calcium carbonate.\",\n \"Cornstarch and the paper component of plastic laminate paper are added as fillers, the components are stirred and mixed, put into the hopper of an extrusion molding machine, kneaded, heated, melted, and pressurized while water is supplied, and then discharged into the air and expanded to obtain a continuous foam.\",\n \"One way to increase the foaming expansion ratio is to add a plastic material that is miscible with the plastic component of the plastic laminate paper.\",\n \"After the plastic material, organic plant component, and inorganic component are kneaded, water is added as a foaming agent, and it is considered important that the various components be uniformly dispersed if a uniform foam is to be obtained.\",\n \"To this end, it is preferable if discarded plastic laminate paper is pulverized and made into pellets in a pelletizer, to which are added the required amounts of plastic, organic plant component, and inorganic component, the components are stirred and kneaded in a mixer, and the resulting composition is supplied to an extrusion molding machine.\",\n \"With this technique, the manufacturing process is characterized in that the method for mixing the materials involves merely mixing the plastic component, plant component (such as cornstarch or paper), and inorganic component.\",\n \"Even with this disclosed technique, there is no special technical description of the above mixing step other than to say that the goal is to obtain a uniform form by merely subjecting raw materials of different melting temperatures and viscosities to a kneading step, and achieving the uniform dispersion of the various components.\",\n \"It is obvious that this disclosed information does not solve the problem related to viscosity and fluidity encountered by the inventors of the present invention.\",\n \"A foam is basically a resource-conserving material in that the amount of materials consumed is extremely small.\",\n \"Foams have excellent adiabatic properties and impact resistance, and are lightweight, which means that they can be used in a wide variety of situations, including applications as packaging materials, and for this reason resin manufacturers and others have conducted intensive research into foams.\",\n \"The flip-side of being lightweight is that foams also have a large volume, so their development requires adequate consideration of their material characteristics as related to recyclability and disposal.\",\n \"As already touched upon in prior art, if a foam could be made from pulverized waste paper or another plant-derived fibrous component or a starch or the like and, if needed, a plastic resin or the like that has been confirmed to be safe to humans and the environment, this would have considerable social significance as well, in view of the high cost of developing new resins and so forth.\",\n \"The present invention solves the problems with viscosity and fluidity inevitably encountered with mixtures of a plurality of different types of polymer compound in the manufacture of a foam by mixing a plastic resin with the above-mentioned natural materials, and establishes a method for manufacturing a foam of uniform quality, with which stable manufacture on an industrial scale is possible.\"\n ],\n [\n \"<SOH> BRIEF DESCRIPTION OF THE DRAWINGS <EOH>FIG.\",\n \"1 is a concept diagram of the manufacturing method of the present invention; FIG.\",\n \"2 is a cross sectional micrograph of the present invention; FIG.\",\n \"3 is a cross sectional micrograph of the present invention; FIG.\",\n \"4 is a micrograph of the interior of the foam of the present invention; FIG.\",\n \"5 is a data table of the results of foam manufacture in the present invention; FIG.\",\n \"6A is a flow state diagram of when the pressure is relatively low in the reservoir in prior art, and FIG.\",\n \"6B is a flow state diagram of slip cone generation when the pressure is relatively high in prior art; FIG.\",\n \"7 is a graph of the viscosity of various kinds of starch in prior art; FIG.\",\n \"8 is a graph of starch viscosity characteristics in prior art; FIG.\",\n \"9 is a graph of elongation viscosity characteristics in prior art; FIG.\",\n \"10 is a flow chart of a foam manufacturing method in prior art; FIG.\",\n \"11 is a simplified diagram of the main parts of a foam manufacturing apparatus in prior art; FIG.\",\n \"12 is a cut-away oblique view of the raw material extrusion die in the manufacture of a foam board in prior art; FIG.\",\n \"13 is a manufacturing state diagram of a foam board in prior art; and FIG.\",\n \"14 is a data table of the results of foam manufacture in prior art.\",\n \"detailed-description description=\\\"Detailed Description\\\" end=\\\"lead\\\"?\"\n ],\n [\n \"TECHNICAL FIELD This invention relates to a method for manufacturing a high-quality foam that can be produced stably on an industrial scale and that affords excellent thermal insulation as a foam with thin cell membranes, which method ensures uniform molten mixture fluidity, that is an essential condition for achieving consistent foam quality, in the manufacture of a foam by mixing starch as one molten polymer compound with another polymer compound having a different melt viscosity, and to a foam manufactured by this method.\",\n \"BACKGROUND ART There have been countless attempts at manufacturing a foam by mixing starch with another plant-derived material or with a plastic resin that would pose no danger of emitting dioxins if incinerated after use, for example, and at utilizing such foams as thermal insulation materials or as cushioning materials or other such shipping and packaging materials, and a number of methods for manufacturing foams have been disclosed.\",\n \"Nevertheless, based on an examination of this disclosed information, it appears that no known technique has been established relating to the problem the present invention is intended to solve.\",\n \"To clarify the problem the present invention is intended to solve, general concepts, beliefs, related examples, and so forth were examined in the technological field to which the present invention is related.\",\n \"It is commonly held that when steam or the like is used as a foaming gas, the material is discharged under pressure from an extrusion molding machine, and reduced-pressure foaming is performed, for instance, a polymer raw material that becomes fluid when the temperature is raised will flow as shown in FIG.\",\n \"6A, with symmetrical flow lines around the center line in the drawing, when the pressure is relatively low, and as the pressure is gradually raised, slip is observed between the material and the walls.\",\n \"At even higher pressures the flow lines begin to be disarrayed, the material is extruded in a spiral shape, and this results in a lack of uniformity in the material.\",\n \"What happens here is called stick and flow, in which the flow of the material comes to a halt (sticking), after which the material starts sliding again (slipping).\",\n \"When this occurs, complete and intermittent slipping over the entire surface of the slip cone shown in FIG.\",\n \"6B has been reported for high density polyethylene and other such polymer materials, and a similar phenomenon is seen not just with polymer compounds but also in the extrusion of bismuth-tellurium and other such metal materials that are thermoelectric semiconductors, for example.\",\n \"This is manifested as cracks in the extruded semiconductor.\",\n \"Various apparatus improvements have been attempted from a mechanical aspect in an effort to eliminate this problem, but the fact is that the fluid properties of polymer compounds are still not fully understood by researchers.\",\n \"The materials that are the subject of the present invention are polymer compounds, and it is necessary to understand this phenomenon.\",\n \"Since the present invention involves a plurality of polymer compounds, however, it is uncertain whether the phenomenon witnessed with a single compound such as polyethylene would be the same, and the problem is even more complicated because we are dealing with a mixture in which the temperature dependency of viscosity and the thermal deformation temperature characteristics vary with the different raw material polymer compounds, which probably makes it even more difficult to understand the situation in terms of fluid dynamics or tribology.\",\n \"The present invention relates to a field of technology in which such scientific understanding is not necessarily perfect, but at the very least, of course, we must know something about the material characteristics of the raw material polymer compounds being used.\",\n \"Let us now give a brief summary of the characteristics of a polypropylene resin, as an example of a plastic resin, and starch, which are believed to be the main contributors to fluidity out of the polymer compound materials used in the present invention.\",\n \"Starch is usually in the form of macromolecular spherulites of a mixture of two homologues with different structures, amylose and amylopectin.\",\n \"The size thereof can vary greatly with the type of starch, but the diameter is said to range from 1 to 20 microns, and the molecular weight up to a few thousand.\",\n \"Amylose is a linear polymer to which D-glucose units are connected by α-1,4 linkages, whereas amylopectin is a branched polymer that includes α-1,6 linkages in addition to α-1,4 linkages, with branching occurring at these sites and the branches connected again with α-1,4 linkages.\",\n \"When starch granules are suspended in water and heated to a certain temperature or higher, the starch granules undergo irreversible swelling and amylose is eluted.\",\n \"This process is called agglutination, and this temperature is referred to as the agglutination temperature.\",\n \"FIG.\",\n \"7 shows published heating and viscosity curves for various kinds of starch (amylograms).\",\n \"The agglutination temperature ranges from 60 to 90° C., and cornstarch is what is cited in the embodiments and so forth of the present invention.\",\n \"FIG.\",\n \"8 also gives published information, and is a comparison of the viscosity behavior of ordinary wheat resulting from agglutination, by variety of glutinous wheat (which has a relatively high amylopectin content), versus that of cornstarch.\",\n \"For this comparison, the temperature of each starch was raised from 34° C. to 94° C. at a rate of 5° C. per minute, after which the starch was cooled at the same rate of 5° C. per minute and its viscosity was measured.\",\n \"Of these varieties, Norin No.\",\n \"61 had the same amylose content of 31.8% as ordinary wheat, and its agglutination temperature was close to that of wheat as shown in FIG.\",\n \"8.The waxy cornstarch used as a control also has a relatively low agglutination temperature, similar to what is shown in FIG.\",\n \"7.The purpose of providing FIGS.\",\n \"7 and 8 here is to show the agglutination temperature of the starch that is one of the polymer compound materials used in the present invention in relation to the fluidity of these materials, and demonstrate that there is a rapid increase in viscosity as soon as the agglutination temperature is attained.\",\n \"The polypropylene resin will now be discussed.\",\n \"Polypropylene is a crystalline thermoplastic resin made by the polymerization of propylene (C3H6), and is hard and tough, with good resistance to moisture, oils, and solvents.\",\n \"Its heat resistance temperature is said to be 170° C. or lower.\",\n \"Also, there is almost no possibility that a linear polymer compound will undergo cyclization during combustion, and these resins, along with polyethylene and the like, are called polyolefin resins and said to be resins that do not produce dioxins or other harmful substances.\",\n \"According to disclosed reference materials from polypropylene manufacturers and so forth, the thermal deformation temperature of polypropylene varies somewhat with the structure of the compound, but is said to be roughly 100 to 120° C. FIG.\",\n \"9 shows an example of the elongation viscosity of the linear (without branches) polypropylene resin used in the examples and so forth of the present invention, and it can be seen that the elongation viscosity at 180° C. quickly disappears in just a few seconds.\",\n \"The reason a foam of a high expansion ratio cannot be produced from a linear polypropylene resin is said to be that the elongation viscosity drops off so quickly and the melt tension is low, and most polypropylene manufacturers produce polypropylene resins having a branched structure by subjecting a linear polypropylene to a special process, and then make a foam with a relatively high expansion ratio from this polypropylene resin with improved elongation viscosity and melt tension.\",\n \"Higher cost is at the present time inevitable with a branched polypropylene because a linear polypropylene must be subjected to a post-treatment step such as electron beam irradiation.\",\n \"When an attempt is made to produce a foam by mixing and melting a starch and a plastic resin such as polypropylene, the difference between the temperature dependence of the viscosity behavior and the melt temperature of the polymer compounds results in complex fluidity of the mixed materials in the mixing, melting, and extrusion steps during manufacture, and it is not hard to imagine the effect that this has on discharge uniformity.\",\n \"Japanese Laid-Open Patent Application H9-111029 discusses a method for manufacturing a loose polypropylene foam by mixing a polypropylene resin and a starch-based additive into a plant-derived foaming agent such as nonfat powdered milk or bean-curd lees, putting this raw material mixture into an extruder comprising a high-temperature heated cylinder, and feeding in water.\",\n \"This publication also discusses a method in which a plastic component, plant-derived foaming agent, and starch-based additive are simply mixed.\",\n \"The inventors of the present invention conducted manufacturing tests in which they used an actual foam manufacturing apparatus similar to that in the disclosed information, used as their raw materials a plastic resin, starch, and pulverized waste paper (as the plant-derived fibrous material), and varied the amount of feed water (the foaming agent), the mixing time and temperature in the manufacturing process, the conditions of the material discharge orifice, and so forth.\",\n \"As a result, with a simple mixture of raw materials as in the disclosed information, perhaps because variance in fluidity could not be suppressed, there was considerably fluctuation in the discharge amount per hour, and there was also variance in the foaming state, so it was found that a foam of stable quality could not be manufactured.\",\n \"The specific manufacturing method and steps here are illustrated in FIG.\",\n \"10.Pulverized paper (such as waste paper) was used as the plant-derived fibrous material, but first the material was finely pulverized in a mixer/cutter type of pulverizer and then in a ball mill type of pulverizer, this pulverized material was sifted through an industrial sieve to obtain a waste paper powder with particles averaging 20 to 30 microns in size, and this powder was used as the waste paper raw material.\",\n \"This waste paper raw material was then mixed with a polypropylene resin to obtain a raw material mixture, to which cornstarch was further added and mixed, and these components were put in a mixing machine and mixed.\",\n \"The sequence of this mixing was also varied so that the waste paper raw material was first mixed with the cornstarch to obtain a raw material mixture, to which the polypropylene resin was added and mixed in a mixing machine.\",\n \"The mixing was also performed at various blend ratios, including those given in the disclosed information.\",\n \"This kneaded raw material was put into,a biaxial extruder and foamed.\",\n \"Specifically, the biaxial extruder shown in FIG.\",\n \"11 comprises a raw material inlet 1, a water inlet 2, cylinders C1 to C5, heaters 3 attached to the various cylinders, a raw material extrusion die (spinneret) 4, and an extrusion opening 5.In manufacturing tests, the settings of the cylinder and extrusion die temperatures was suitably varied.\",\n \"The test was conducted with the water supply varied between 10 and 30 liters per hour.\",\n \"To evaluate the foam state and foam variance, a string-like piece of foam extruded and foamed from an extrusion die with a diameter of 2 mm was cut up by an automatic cutter set to a specific rotating speed, 50 pieces were randomly sampled from among the total of 4200 pieces of cut foam produced in one minute, whose combined length was 120 m, the length and diameter of the cut foam pieces were measured, and the extent of foaming was visually evaluated on a ten-point scale.\",\n \"The mixing ratios of the materials for which the results are shown in FIG.\",\n \"14 were 25% (weight ratio) pulverized waste paper, 30% starch, and 45% polypropylene resin.\",\n \"As indicated by these results, the foam manufactured by the mixing process described above exhibited considerable variance in all measurement categories.\",\n \"The length of cushioning material cut at regular time intervals varied from a minimum of 1.9 cm to a maximum of 8.5 cm.\",\n \"The most serious problem was variance in the extent of foaming.\",\n \"The ten-point scale assumes 10 to be the ideal foaming state, but because there was variance in the foaming, the cushioning material varied from thin to thick.\",\n \"This indicates nothing other than a change over time in the fluidity of the raw material mixture during mixing and melting inside the cylinders.\",\n \"This is similar to the stick and flow phenomenon described above, but it is unclear whether it occurred because a slip cone was formed, or just because of local variance in the fluidity of the molten mixture.\",\n \"Admixed in this molten mixture was pulverized waste paper, whose characteristics remained almost unchanged over the temperature range in this process.\",\n \"This pulverized paper consists of a fibrous material with a microscopic length of anywhere from a few microns to about 100 microns, which is stirred and mixed in the biaxial extruder, and it seems that nuclear cells that will serve as the nuclei for cells are produced here and there in an agitated fluid in which polymer compounds of different viscosity are admixed.\",\n \"In this situation it is doubtful whether a nice slip cone is being formed as in the above-mentioned reports on polyethylene resins.\",\n \"On the contrary, it seems likely that this nucleus formation is occurring as a result of local variance in the fluidity of the molten mixture.\",\n \"At this point we will briefly discuss the correlation between the viscosity of the above-mentioned starch and polypropylene resin and the fluidity of the mixture of polymer compounds, since such a discussion will probably further clarify the points involved in the problems which the present invention is intended to solve and the means used to solve the problems.\",\n \"A capillary type of viscosity measurement apparatus is the most commonly used in the measurement of the viscosity of polymers.\",\n \"A sample is extruded from a capillary tube by applying pressure, and the viscosity is measured by measuring the difference in the applied pressure at a constant flow rate per unit of time.\",\n \"We will let L be the length of the capillary tube, and r be its radius.\",\n \"The change over time in the amount of sample flowing out of the capillary tube is found for when pressure is applied such that the sample contained in the reservoir flows at a specific rate, and this is termed Q (cm3/sec).\",\n \"A relationship of γ=4Q/πr3 exists between the shear rate γ (sec−1) and the sample flowing through the capillary tube, and the shear stress τ (N/cm2) on the inner walls of the capillary tube is expressed as τ=rP/2L.\",\n \"The viscosity μ (N·sec/cm2) is defined from the shear rate y and the shear stress τ as: μ=τ/γ.\",\n \"If we assume that a starch and a polypropylene resin are mixed and the thermal characteristics of each are independently manifested, the intrinsic viscosity should be exhibited in the starch at the agglutination temperature and in the polypropylene resin at the thermal deformation temperature, with the viscosity subsequently varying according to temperature changes.\",\n \"For instance, if the agglutination temperature of the starch is reached before the thermal deformation temperature is reached, the viscosity of the starch increases suddenly, and the entire system is governed by the viscosity of the starch.\",\n \"If the thermal deformation temperature of the polypropylene resin is never reached, the polypropylene might have some kind of effect as an admixture, but the viscosity μ of the system will be relatively stable.\",\n \"In other words, this is a state in which the fluidity of the system is governed by the viscosity of the starch, even though the system is a mixture.\",\n \"However, this never happens for the purposes of manufacturing a foam.\",\n \"Still, if the temperature continued to rise, it would eventually reach the thermal deformation temperature of the polypropylene resin, resulting in a fluid having both the viscosity of polypropylene and the viscosity of the starch, which varies with time and temperature.\",\n \"It is very difficult to know what the mixing state is in a mixed fluid such as this.\",\n \"Measurement was made at a certain specific temperature with the above-mentioned viscosity measurement method, but since the mixing is not completely uniform in a mixing state of different viscosities, the local shear rate γ should vary.\",\n \"Furthermore, the shear stress τ on the inner walls of the capillary tube also varies according to the local mixing state.\",\n \"In other words, the viscosity μ of a mixture of polymer compounds changes with how uniform the mixing is, and if we look at the viscosity definition of the above formula, we can see that there is extreme instability in which both the denominator and the numerator vary.\",\n \"Put another way, there is no viscosity that governs the mixture as a system, and instead the system is in a state of constant change.\",\n \"Japanese Laid-Open Patent Application 2000-143869, which is another source of disclosed information, discloses a technique of manufacturing a foam by pulverizing a plastic laminate and mixing with polypropylene, cornstarch, and calcium carbonate.\",\n \"With this technique, the foaming expansion ratio is about 10 times with just the resin that is admixed in the laminate paper, and the admixture of another plastic is proposed in order to increase the foaming expansion ratio.\",\n \"Close examination of this technique reveals that, as seen in the embodiments and examples of the invention, a foamable composition is made up of plastic from pulverized laminate paper and added calcium carbonate.\",\n \"Cornstarch and the paper component of plastic laminate paper are added as fillers, the components are stirred and mixed, put into the hopper of an extrusion molding machine, kneaded, heated, melted, and pressurized while water is supplied, and then discharged into the air and expanded to obtain a continuous foam.\",\n \"One way to increase the foaming expansion ratio is to add a plastic material that is miscible with the plastic component of the plastic laminate paper.\",\n \"After the plastic material, organic plant component, and inorganic component are kneaded, water is added as a foaming agent, and it is considered important that the various components be uniformly dispersed if a uniform foam is to be obtained.\",\n \"To this end, it is preferable if discarded plastic laminate paper is pulverized and made into pellets in a pelletizer, to which are added the required amounts of plastic, organic plant component, and inorganic component, the components are stirred and kneaded in a mixer, and the resulting composition is supplied to an extrusion molding machine.\",\n \"With this technique, the manufacturing process is characterized in that the method for mixing the materials involves merely mixing the plastic component, plant component (such as cornstarch or paper), and inorganic component.\",\n \"Even with this disclosed technique, there is no special technical description of the above mixing step other than to say that the goal is to obtain a uniform form by merely subjecting raw materials of different melting temperatures and viscosities to a kneading step, and achieving the uniform dispersion of the various components.\",\n \"It is obvious that this disclosed information does not solve the problem related to viscosity and fluidity encountered by the inventors of the present invention.\",\n \"A foam is basically a resource-conserving material in that the amount of materials consumed is extremely small.\",\n \"Foams have excellent adiabatic properties and impact resistance, and are lightweight, which means that they can be used in a wide variety of situations, including applications as packaging materials, and for this reason resin manufacturers and others have conducted intensive research into foams.\",\n \"The flip-side of being lightweight is that foams also have a large volume, so their development requires adequate consideration of their material characteristics as related to recyclability and disposal.\",\n \"As already touched upon in prior art, if a foam could be made from pulverized waste paper or another plant-derived fibrous component or a starch or the like and, if needed, a plastic resin or the like that has been confirmed to be safe to humans and the environment, this would have considerable social significance as well, in view of the high cost of developing new resins and so forth.\",\n \"The present invention solves the problems with viscosity and fluidity inevitably encountered with mixtures of a plurality of different types of polymer compound in the manufacture of a foam by mixing a plastic resin with the above-mentioned natural materials, and establishes a method for manufacturing a foam of uniform quality, with which stable manufacture on an industrial scale is possible.\",\n \"DISCLOSURE OF THE INVENTION The present invention provides a high-quality foam that can be produced stably on an industrial scale and that affords excellent thermal insulation as a foam with thin cell membranes.\",\n \"Also, efficiently utilizing a starch (a natural polymer compound) has great industrial significance in that even a linear plastic resin that is inexpensive and has low elongation viscosity can be used to manufacture a foam with a high foaming expansion ratio.\",\n \"Furthermore, the present invention provides a novel way of thinking about the scientific phenomena of agglutination and mixed fluids of a plurality of polymer compounds.\",\n \"Also, with the present invention, in the manufacture of a foam of a plastic resin containing as compositional components pulverized paper, pulverized wood, or another such plant-derived fibrous material and starch, there is a step of adding water to the plant-derived fibrous material and starch and mixing the components to manufacture pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination, and a step of foaming the pellets by mixing them with a plastic resin, and included in between these two steps is a step of drying the pellets at a temperature not higher than the starch agglutination temperature.\",\n \"The result is that it is possible to manufacture a foam of uniform quality, and one that has a high foaming expansion ratio even though it is made from an inexpensive linear plastic resin with low elongation viscosity.\",\n \"Also, with the present invention, the pellets are dried for a relatively long time at a low temperature of 50° C. or lower, the advantage of which is a higher foaming expansion ratio.\",\n \"Also, with the present invention, in the manufacture of a foam of a plastic resin containing as compositional components pulverized paper, pulverized wood, or another such plant-derived fibrous material and starch, there is a step of adding water to the plant-derived fibrous material and starch and mixing the components to manufacture pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination, and a step of foaming the pellets by mixing them with a plastic resin, and included in between these two steps is a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized paper as a compositional component, and the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, or the pulverized paper being made from high-grade waste paper or from printing scraps or waste of postal cards, stamps such as postage stamps, currency, and so forth.\",\n \"The result is that it is possible to manufacture a foam with a high foaming expansion ratio from pulverized paper, and particularly high-grade waste paper, and this foam can be manufactured inexpensively, at a uniform level of quality, and will be even safer.\",\n \"Also, with the present invention, in the manufacture of a foam of a plastic resin containing as compositional components pulverized paper, pulverized wood, or another such plant-derived fibrous material and starch, there is a step of adding water to the plant-derived fibrous material and starch and mixing the components to manufacture pellets in which starch molecular spheres are agglomerated to each other or to the plant-derived fibrous material without the starch undergoing agglutination, and a step of foaming the pellets by mixing them with a plastic resin, and included in between these two steps is a step of drying the pellets at a temperature not higher than the starch agglutination temperature, the plant-derived fibrous material comprising pulverized wood as a compositional component, the plastic resin comprising a polyolefin resin such as a polypropylene resin or polyethylene resin as a compositional component, or the pulverized wood being of reed or the like, the advantage of which is that a foam of particularly high quality can be manufactured from pulverized wood, and reed in particular.\",\n \"Also, with the present invention, in the above-mentioned foam manufacturing method, the plant-derived fibrous material component accounts for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %, which allows a foam with a relatively high expansion ratio to be manufactured.\",\n \"Also, the present invention provides relatively inexpensively a foam manufactured by the manufacturing methods described above, having a high foaming expansion ratio and excellent thermal insulation performance.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS FIG.\",\n \"1 is a concept diagram of the manufacturing method of the present invention; FIG.\",\n \"2 is a cross sectional micrograph of the present invention; FIG.\",\n \"3 is a cross sectional micrograph of the present invention; FIG.\",\n \"4 is a micrograph of the interior of the foam of the present invention; FIG.\",\n \"5 is a data table of the results of foam manufacture in the present invention; FIG.\",\n \"6A is a flow state diagram of when the pressure is relatively low in the reservoir in prior art, and FIG.\",\n \"6B is a flow state diagram of slip cone generation when the pressure is relatively high in prior art; FIG.\",\n \"7 is a graph of the viscosity of various kinds of starch in prior art; FIG.\",\n \"8 is a graph of starch viscosity characteristics in prior art; FIG.\",\n \"9 is a graph of elongation viscosity characteristics in prior art; FIG.\",\n \"10 is a flow chart of a foam manufacturing method in prior art; FIG.\",\n \"11 is a simplified diagram of the main parts of a foam manufacturing apparatus in prior art; FIG.\",\n \"12 is a cut-away oblique view of the raw material extrusion die in the manufacture of a foam board in prior art; FIG.\",\n \"13 is a manufacturing state diagram of a foam board in prior art; and FIG.\",\n \"14 is a data table of the results of foam manufacture in prior art.\",\n \"BEST MODE FOR CARRYING OUT THE INVENTION The present invention will now be described through reference to the drawings.\",\n \"It was discussed above that it is difficult to mix and melt a plurality of polymer compounds having different viscosities and then extrude and foam this mixture while keeping the fluidity as uniform as possible, if the raw materials are merely mixed.\",\n \"For the present invention, investigation focused on the mixing of a pulverized paper component and a starch.\",\n \"The present invention involves creating this mixture by an entirely new idea and manufacturing method that did not exist until now, supplying the mixture thus produced and a plastic resin to a biaxial extruder along with water, and mixing and melting the components to manufacture a foam of uniform quality.\",\n \"The means for accomplishing this will now be described in specific terms.\",\n \"Even if the raw material mixing step involves first mixing the pulverized paper with the starch, and then mixing in the propylene or other plastic resin, unless a special method is employed for the first mixing step, uniform fluidity cannot be ensured in the second mixing step that occurs when the components are put into the biaxial extruder, so even though the steps are separate, they will be referred to simply as mixing.\",\n \"The phrases “mere mixing” and “merely mixing” as used in this Specification are in every case defined in this way.\",\n \"The starch used in the embodiments is cornstarch, whose agglutination temperature is generally held to be between 80 and 90° C., but since agglutination actually begins as soon as the temperature goes over 60° C., it was found that the agglutination temperature varies with the processing performed by the starch manufacturer.\",\n \"Further, since the thermal deformation temperature of a plastic resin is 100 to 120° C., the cornstarch agglutinates first and the viscosity increases suddenly, so particularly when the starch is contained in a sufficiently large quantity (weight ratio), it is almost unavoidable that the melt mixing state with the polypropylene resin will be heterogeneous.\",\n \"This applies not only to polypropylene, but also to polyethylene and other resins, as long as the thermal deformation temperature of the plastic resin is higher than the agglutination temperature of the starch.\",\n \"It is also obvious that the effect the starch has on the fluid will be completely different if the amount of starch is far too low.\",\n \"However, the linear polypropylene resin used in the embodiments of the present invention has a low viscous stretching strength and stretching viscosity, which prevents cell growth, and it is said that with a single resin the foaming expansion ratio will be only a few times.\",\n \"The blend proportion of starch and the function thereof are important if a linear polypropylene resin is to be used to manufacture a foam with a high foaming expansion ratio.\",\n \"In an effort to solve the problems related to fluidity and viscosity that occur with a mixture having a sufficiently high starch content, the inventors researched the question of whether the agglomeration effect of the starch molecules on the surface of pulverized paper could be effectively utilized in the manufacture of a mixture of pulverized paper and starch.\",\n \"This idea was the starting point for the present invention, resulting in an extremely useful invention and at the same time yielding a discovery that is significant in terms of understanding the phenomena of agglomeration and colloid formation, which still leave many scientifically unanswered questions.\",\n \"Starches usually will not dissolve in water.\",\n \"They disperse in a colloidal state.\",\n \"The size of molecular spherulites of starch varies with the type of starch, but is said to be from 1 to 200 microns.\",\n \"In the examples of the present invention, the pulverized paper and starch are stirred and mixed for approximately 60 minutes in a ribbon mixer, after which 1.5 liters of water is added to 3 kg of mixture and kneaded for about 5 minutes in a batch kneader.\",\n \"This kneaded sol is granulated in a wet granulator.\",\n \"These granules are called pellets.\",\n \"After this a dryer is used to perform static drying for approximately 20 hours at 40° C. The pellets are stirred around once or twice during this time to ensure uniform drying.\",\n \"The highest temperature throughout the kneading and drying steps is 40° C., at which no agglutination of the starch occurs at all.\",\n \"If the drying temperature is far too high, the starch molecular spheres of the pellets may undergo partial agglutination.\",\n \"As discussed below, temperature control is very important during the drying process, since it can affect fluidity or even affect the function of the starch as a binder, and hinder the formation of thin cell membranes.\",\n \"The above-mentioned pulverized paper is high-grade waste paper or discarded postal cards (including postal cards made of materials equivalent to that of postal cards), postage stamps or other such stamps, currency, and so forth, or printing scraps thereof.\",\n \"The high-grade waste paper referred to in this Specification must meet three conditions: (1) the fiber content must be at least about 8 times the total content of standard waste paper, (2) the material must be safe enough that any hazardous substances such as formaldehyde it contains pass the standards set forth in the test methods of the Japanese “Harmful Substance-Containing Household Article Control Act,” (3) no chlorine or other bleaching agents must be used in the processing of this waste paper.\",\n \"An example of the above-mentioned pulverized wood is of reed (a perennial grass of the Gramineae family).\",\n \"Wood of a substance equivalent to the above-mentioned reed is also encompassed.\",\n \"The particles of pulverized paper or pulverized wood average about 10 to 20 microns in thickness and a few dozen microns in length.\",\n \"In the case of starch molecular spheres about 20 microns in diameter, the surface area of these particles is sufficient for about 10 to 20 of these spheres to adhere.\",\n \"The forces at work in agglomeration are said to include the van der Waals force, electrostatic attraction, and the solid crosslinking force, but with molecules adhering to the surface of pulverized paper, the force of mechanical adhesion to microscopic protrusions on the surface cannot be overlooked, and the adsorption strength of starch molecular spheres adhering to the surface is believed to be high.\",\n \"Judging from the numbers of pulverized paper particles and starch molecular spheres, a number of layers of starch molecular spheres are agglomerated surrounding starch molecular spheres adsorbed to the pulverized paper surface.\",\n \"Table 1 shows the results of putting specific amounts of two types of pellets that had undergone this agglomeration processing and the starch used as the raw material into a beaker, adding water, and heating and stirring while observing how the state changes with temperature.\",\n \"TABLE 1 Temperature (° C.) Starch alone Pellets A Pellets B 50 no agglutination 75 agglutination 100 125 no agglutination 150 no agglutination agglutination Pellets A were produced by producing a raw material mixture of equal weights of pulverized paper and starch, kneading this with water in a weight ratio of 2 to 1, and extruding this product at a granulation and extrusion orifice diameter of 1 mm to produce pellets, then drying these for 20 hours at 40° C. Pellets B were produced under exactly the same conditions, except that the granulation and extrusion orifice diameter was 2 mm.\",\n \"As shown in Table 1, with starch alone, agglutination begins and viscosity rises suddenly at 60° C. or higher, but with a starch that has agglomerated on the pulverized paper surface, no agglutination occurs at this temperature, although there is a colloidal state in which agglomerated starch on the pellet surfaces is partially dispersed.\",\n \"The only difference between pellets A and B is the granulation orifice diameter.\",\n \"The agglomeration force and the breakdown thereof are not yet fully understood, but one cannot help but think that the difference is that when the granulation orifice diameter is 1 mm, the extrusion pressure increases and the agglomeration force grows.\",\n \"For example, there has been research indicating that ceramic firing varies with differences in the agglomeration force balance due to how the material components are agglomerated or with control of the agglomeration structure, and the inventors of the present invention have observed in investigations leading up to the present invention that agglomerated starch molecular spheres are eluted when the pellet drying temperature is varied between 30 and 50° C., that is, that there is a subtle difference in the colloidal state, which makes it extremely interesting that slight differences in conditions can produce differences in agglomeration.\",\n \"It has been mentioned that many parts of the chemistry of agglomeration are still unclear, but in any case the fact that the agglomerated starch of pellets exhibits behavior completely distinct from that of the agglutination of starch alone is important when considering the fluidity of a mixture containing a plastic resin.\",\n \"The present invention involves mixing a plastic resin with agglomerated pellets exhibiting the above properties, putting this mixture into a biaxial extruder or the like, and manufacturing a foam.\",\n \"When a polypropylene resin is used as the plastic resin, since the thermal deformation temperature is only about 20° C. higher with respect to cornstarch, unless there is a sharp increase in agglutination and viscosity of the starch in this temperature region, the fluidity of the polypropylene is the main factor in determining the fluidity of the entire mixture.\",\n \"This the problem to be solved, namely, the above-mentioned governing viscosity and fluidity of the system.\",\n \"When the agglomerated pellets and the plastic resin are put into the biaxial extruder and while the mixture is proceeding to the discharge orifice, it is believed that the pellets begin to melt while containing relatively small amounts of starch colloid dispersed from the pellets, resulting in a state of being surrounded by polypropylene.\",\n \"It is surmised that agglutination of the starch dispersed from the pellets occurs in the high-temperature cylinder at the end, and finally the starch molecular spheres near the pulverized paper surface undergo agglutination.\",\n \"This incremental agglutination theory would seem to be the correct view, judging from the shape of the foam as discussed below.\",\n \"FIG.\",\n \"2 is a cross sectional micrograph of a foam produced using a linear polypropylene (one with no branches), pellets of cornstarch, and pulverized paper produced as above.\",\n \"The compositional ratios (in weight percent) were 28%, 24%, and 48%, respectively.\",\n \"It can be seen right away that the foaming expansion ratio is very high despite the use of a linear polypropylene, with which it is held that a foam with a high foaming expansion ratio cannot be obtained because the stretch viscosity is low.\",\n \"Naturally, the higher the foaming expansion ratio, the thinner the cell membranes, and the membrane thickness is just a few microns.\",\n \"With a linear polypropylene alone, it is said that the foaming expansion ratio will only be about 2 to 3 times, and cross sectional views provided by manufacturers reveal the membrane thickness to be several dozen microns.\",\n \"Taking a look at why, in contrast, a foam with such thin membranes is formed from a polypropylene with the same low stretch viscosity when mixed with pulverized paper and cornstarch as in the present invention is also important in terms of understanding the incremental agglutination of starch and how the fluidity of the system is governed.\",\n \"When a foam is manufactured from a single plastic resin, small melting-resistant particles commonly called frothers are added, mixed, and stirred, and it is said that a foam is produced through the growth of microscopic bubbles called nuclear cells, which are generated as a result of the different phases.\",\n \"The growth of these cells is said to be determined exclusively by the stretch viscosity and surface tension of the resin.\",\n \"However, it would seem to be impossible to explain the formation of the thin membranes in the present invention by this widely-held theory alone.\",\n \"We probably have no choice but to conclude that the situation is somewhat different for the foam formation mechanism of a mixture that contains relatively large amounts of polymer materials of different viscosities and that contains pulverized paper, whose properties and shape remain basically unchanged at the temperatures in all of the steps.\",\n \"Nuclear cells are generated when the flow of a fluid with different viscosities becomes turbid.\",\n \"It is believed that nuclear cells are readily generated in many locations during the foam manufacturing process of the present invention, in which a fibrous material is admixed, and that the number of these cells is extremely large.\",\n \"When these countless nuclear cells are discharged under reduced pressure from a biaxial extruder, they grow all at once.\",\n \"As said in the case of a single resin comprising linear polypropylene, if the stretch viscosity of the resin is low, most of these nuclear cells will not grow completely, resulting in cell clusters, and as a result the foaming ratio is lower and the cell membranes are thicker.\",\n \"However, with the foam of the present invention containing a starch, it seems that a major role is played by the starch, in which changes in the agglutination state arise from condensation, and the pulverized paper fibrous material.\",\n \"The amount of starch component that mixes with the molten polypropylene is relatively small compared to the amount of raw materials added, and it is reasonable to conclude that most of the starch stops around the pulverized paper.\",\n \"There should be a local increase in viscosity as a result of the agglutination of starch eluted while the polypropylene is molten, but since the amount is not very large, the mixture as an entire system seems to exhibit relatively stable fluidity.\",\n \"As discussed previously regarding the colloid formation that accompanies an increase in temperature, the agglomerated starch adhering around the pulverized paper is not dispersed and eluted all at once, but instead gradually disperses from the agglomeration surface.\",\n \"Near the last stage of the biaxial extruder, that is, the discharge orifice, all of the starch is believed to be agglutinated, but pellets are present in dispersed form in the completely melted polypropylene resin fluid, and the concentration distribution of amylose in the agglutinated starch is believed to have a gradient going outward from around the pulverized paper, that is, toward the main polypropylene fluid.\",\n \"It is surmised that, despite their low stretch viscosity, the polypropylene membranes that have joined together with the partially agglutinated starch are given a cushioning effect by the elasticity and stretchability of the agglutinated starch surrounding the pulverized paper, and at the same time a balance is achieved in the extremely thin membrane state through the pressure of the adjacent cells that are similarly undergoing rapid growth.\",\n \"The importance of temperature management in the pellet drying process has already been discussed, but if the pellets end up being partially agglutinated in the drying process, this will affect the cushioning effect resulting from the elasticity and stretchability of the starch.\",\n \"FIG.\",\n \"3 is a photograph of a foam that has been cut diagonally.\",\n \"A relatively thick film can be seen surrounding the pulverized paper fibrous material, and we see that there is a continuous reduction in thickness.\",\n \"FIG.\",\n \"4 is a photograph at 10,000-times magnification of the interior of a foam, taken with a scanning electron microscope.\",\n \"A film in which tiny holes a few dozen to a few hundred nanometers in size can be seen is formed between two objects that seem to be fibrous material about 1 micron in size, and that look thick in the photograph.\",\n \"This film is not a cell membrane, but these microscopic observation results seem to bear out the foam membrane formation mechanism of the present invention that has been inferred up to now.\",\n \"As expected, the primary means for solving the problem of the present invention does indeed solve the problem of too much variance in the raw material mixture inside the biaxial extruder by making use of pellets in which the agglomeration force of starch and pulverized paper is put to suitable use, but at the same time the function of starch as a binder, which up to now had only been described in vague terms, becomes more clear, and this is a groundbreaking discovery that is the key to forming extremely thin cell membranes.\",\n \"As discussed above, an embodiment of the present invention relates to the flow stability of a mixed molten fluid, which is a problem in the manufacture of a foam of uniform quality using a plurality of polymer compounds with different melting temperatures or viscosities.\",\n \"The components that make up the foam consist of a pulverized fibrous material that will not melt at the temperatures encountered during manufacture, such as wood or paper whose main component is cellulose, lignin, or the like; starch, which is necessary as a binder for the fibrous material and the plastic resin; a plastic resin having a thermal deformation temperature higher than the agglutination temperature of the starch; and water or the like as a foaming agent.\",\n \"Furthermore, the present invention is the result of a finding that confirms the correctness of the theory based on typical experiments and the theory developed from the results of a number of experiments relating to the function of agglomerates in mixed fluids in which a plastic resin is melted, and that agglomerations of starch molecular spheres are produced, the core of which is made up of starch, which undergoes a major thermal reaction (a phenomenon called agglutination) at the lowest temperature of any of the constituent components other than water, and a pulverized fibrous material of paper, wood, or the like that has substantially no temperature dependence in terms of the fluidity of the mixture, and the reactivity of the agglomerated starch with respect to temperature is different from that of the raw material starch.\",\n \"Therefore, in the foam constituent raw materials mentioned in the test and embodiments cited in this Specification, except for the starch, the fibrous material is not limited to pulverized paper as long as the thermal change defined in the present invention does not occur within the temperature range encountered in the foam manufacturing process.\",\n \"Similarly, a linear polypropylene resin, which is an inexpensive and readily available raw material, is used in the present invention as a typical example of a safe plastic resin that does not give off dioxins or other such harmful substances when disposed of and incinerated, but the plastic resin is not limited to a polypropylene resin as long as it has fluidity and stretchability suited to the manufacture of a foam and has a thermal deformation temperature higher than the agglutination temperature of the starch.\",\n \"Also, water is used as the medium in the present invention to manufacture an agglomerate of starch by adsorbing starch molecular spheres to the plant-derived fibrous material that serves as the core in the present invention, but there are other methods for producing an agglomerate as well, such as causing the substance particles to collide with each other without the use of water, so the agglomerated starch of the present invention may be obtained by any method as long as it yields an agglomerate that exhibits the behavior described in detail in the present invention when melted along with other polymer compounds.\",\n \"For the same reason, regarding the foaming method, steam foaming in which water serves as the foaming agent is performed in the present invention, and to this end the tests and examples described herein make use of a biaxial extruder, but the present invention is not limited to this.\",\n \"Furthermore, a foam of uniform quality can be manufactured in the present invention by having the plant-derived fibrous material (pulverized paper or pulverized wood) component account for 20 to 80 wt %, the starch component for 10 to 30 wt %, and the plastic resin component for 10 to 50 wt %.\",\n \"EXAMPLES Example 1 Example 1 relates to a cushioning material obtained by cutting a string of foam into pieces.\",\n \"This cushioning material is used to protect objects during shipping.\",\n \"A specific manufacturing method and steps will be described through reference to the flow chart of FIG.\",\n \"1.Pellet Production Step Using postal card waste paper as the plant-derived fibrous material, first the paper was coarsely pulverized in a mixer/cutter type of pulverizer, and then it was finely pulverized in a ball mill type of pulverizer.\",\n \"This product was sifted through an industrial sieve to obtain a waste paper raw material of the required size.\",\n \"Next, this waste paper raw material was mixed in an amount of 40% (weight ratio) with cornstarch in a weight ratio of 60% for 60 minutes in a ribbon mixer.\",\n \"3 kg of this mixture was combined with 1.5 liters of water and kneaded for 5 minutes in a batch kneader.\",\n \"Pellets were produced in a wet granulator and allowed to dry for 12 hours in a dryer set at 50° C., after which the pellets were stirred around and dried for another 8 hours to ensure even drying.\",\n \"Biaxial Extrusion Step The dry pellets were mixed with a linear polypropylene resin (as the plastic resin component), a suitable amount of water was supplied, and the components were kneaded well.\",\n \"This kneaded raw material was fed into a biaxial extruder A and foamed.\",\n \"The biaxial extruder shown in FIG.\",\n \"11 comprises a raw material inlet 1, a water inlet 2, cylinders C1 to C5, heaters 3 attached to the various cylinders, a raw material extrusion die (spinneret) 4, and an extrusion opening 5.In manufacturing tests, the settings of the cylinder and extrusion die temperatures was suitably varied.\",\n \"The test was conducted with the water supply varied between 10 and 30 liters per hour.\",\n \"These results are given in FIG.\",\n \"5.The results from FIG.\",\n \"5 are also given in Table 2 in comparison with the results from FIG.\",\n \"14 (prior art).\",\n \"TABLE 2 Comparison table Simple Subject Compared category Pellets1 mixture2 Unit Average length 5.602 4.874 length (cm) Size of range 2.5 6.6 (cm) Maximum length 6.6 8.5 (cm) Minimum length 4.1 1.9 (cm) Standard 0.634 1.923 deviation Unit Average length 2.074 1.886 length (cm) Size of range 0.4 1.4 (cm) Maximum length 2.3 2.4 (cm) Minimum length 1.9 1.0 (cm) Standard 0.110 0.328 deviation [1present invention, 2prior art] It can be seen that there is far less variance in the foam manufactured by the technique of the present invention than in the foam manufactured by prior art.\",\n \"Also, 29 out of 50 samples were within ±10% of the average length with the present invention, whereas the number was only 4 with the prior art.\",\n \"What is even more noteworthy is the difference in regard to diameter.\",\n \"In Table 2, the difference in the variance in foaming extent in FIGS.\",\n \"5 and 14 is expressed directly as the size of the range of variance in diameter.\",\n \"The uniformity of the extent of foaming and the small variance in diameter and length achieved with the technique of the present invention make it possible to manufacture a foam board, which was impossible in terms of quality with prior art.\",\n \"Example 2 Next we will describe the manufacture of a foam board by proceeding through the same steps as in Example 1, but with the extrusion die (spinneret) of the biaxial extruder changed.\",\n \"The extrusion die (spinneret) used in the prior art and Example 1 was intended for manufacturing a foam string, being equipped with two discharge orifices of 2 mm a suitable distance apart, but as shown in FIGS.\",\n \"12 and 13, the structure of an extrusion die for manufacturing a foam board is provided with numerous 2-mm discharge orifices and has a box-like area to the rear thereof.\",\n \"A foam board can be manufactured by passing the material through this die.\",\n \"Foam boards can be applied to the cardboard insides of shipping containers that need to offer thermal insulation.\",\n \"After use, the insulating foam boards can be removed, and can be treated as waste material that will not give off dioxins or other harmful substances when burned.\",\n \"This makes such boards very useful as a thermal insulation material.\",\n \"Since the size of the cardboard container is set, if the dimensional precision of the foam board is off by too much, the board will be too long to fit into the container, or it may be too short, so that it does not stay pressed against the cardboard container and falls over, or there may be gaps in the corners, which compromises the thermal insulation performance of the container, so automatic dimensional precision is important from the standpoint of industrial production.\",\n \"In view of this, we wanted to test how much variance in cut length there was in the foam board of the present invention with respect to the cut length target, and to confirm the foam quality, and therefore examined the thermal insulation of a container in which foam boards were actually installed inside a cardboard container.\",\n \"Table 3 shows the cutting variance test results.\",\n \"In approximately 2 hours of manufacture, a foam board with an overall length of as large as 110 meters was manufactured, with the cutting target set at 1000 mm.\",\n \"Therefore, 110 foam boards cut to 1000 mm were produced.\",\n \"In this test, 20 random samples were collected and measured for length.\",\n \"TABLE 3 Variation in length of foam boards Sample No.\",\n \"Length (cm) 1 100.5 2 100.1 3 100.3 4 99.9 5 100 6 100 7 100.6 8 100.3 9 99.7 10 100 11 100.1 12 100.8 13 100.2 14 99.5 15 99.8 16 100 17 99.8 18 100.3 19 100.3 20 100.1 Average 100.115 Standard deviation 0.311659 As a result, the average length was 100.115 cm, the maximum length was 100.6 cm, and the minimum length was 99.5 cm, giving a standard deviation of 0.312, which means the results were extremely stable.\",\n \"Thus, a foam board could be molded, which could not be obtained with prior art.\",\n \"What is even more interesting is that while the average length of one of the foam strings produced by the technique of the present invention shown in Table 2 of Example 1 was 5.6 cm and the standard deviation was 0.634, but the average length of the foam boards was 100.1 cm, with a standard deviation of only 0.312.With the foam strings, the time it took for a foam of 5.6 cm to be produced was calculated to be a mere 0.028 second.\",\n \"By contrast, with the boards, the foam discharge was slowed by the difference in pressure arising from the shape of the extrusion die (spinneret).\",\n \"The time it took to produce a foam board with a length of 1 meter was approximately 1.2 seconds.\",\n \"In other words, this indicates that inside the biaxial extruder the fluidity fluctuates from instant to instant (0.028 second), but when viewed at a time of 1.2 seconds the fluidity of the system is extremely stable.\",\n \"This proves quite conclusively the correctness of the theory about the fluidity of a composite polymer compound discussed in detail in the prior art section of this Specification, and the correctness of the approach of the method for solving the targeted problem in the present invention.\",\n \"Next, we examined the insulation performance of a cardboard container in which these foam boards were used.\",\n \"For this evaluation we made a comparison with a foamed polystyrene container, which is the most common type of insulating container in use today.\",\n \"The containers used here were as follows.\",\n \"1.Cardboard Container in Which Foam Boards Were Used This was a cardboard container lined on the insides with insulating foam boards (panels) produced according to the present invention using a waste paper component.\",\n \"Container outside dimensions: 290 mm wide×205 mm deep×222 mm high Insulation thickness: 20 mm of foam board Container inside dimensions: 245×160×170 mm 2.Commercially Available Foamed Polystyrene Container Container outside dimensions: 293 mm wide×215 mm deep×220 mm high Insulation thickness: 25 mm Container inside dimensions: 245×165×175 mm Measurement Principles and Method: 1) The container is placed under constant ambient temperature conditions, and then heated with a heater/fan unit inside the container.\",\n \"2) 2.\",\n \"[sic] Power is applied to the heater unit until there is a sufficient temperature differential between the container ambient temperature and the average temperature at three points inside the container.\",\n \"3) Enough time is allowed to pass for the temperature inside the container to become constant (referred to as a steady state).\",\n \"This means that the temperature inside the container and the ambient temperature (constant) reach a state of equilibrium via the insulated container.\",\n \"4) The adiabatic coefficient (or heat leakage coefficient) of the container per 1° C. temperature differential is calculated from the power (W) applied to the heater unit and the difference between the ambient temperature and the temperature inside the container in a steady state.\",\n \"This is expressed as W/° C. Measurement was conducted by the following method.\",\n \"A temperature measurement jig was placed in the approximate center of the container, the internal[5] height of the container was divided into four equal parts, and the temperature at three height points thereof was measured with a copper constantan thermocouple.\",\n \"Variance in the temperature inside the container was reduced by the fan of the heater unit.\",\n \"The ambient temperature was set by the controller of a climatron or a thermostatic tank, but the reliability of the temperature measurement was enhanced by measuring the temperature near the container with a constantan thermocouple to confirm how well it matched the measured temperature inside the container.\",\n \"The heater unit consisted of a heater mounted on a heat sink, and a fan used to diffuse the heat radiated from the heat sink.\",\n \"The heater and the fan were driven by separate DC power supplies, the voltage and current of each were measured, the total applied power was calculated, and this was termed the amount of heat generated inside the container.\",\n \"Thermal insulation performance was measured at two locations externally, and are termed measurement test sites A and B.\",\n \"These results are given in Table 4.In all of these measurements, the cardboard container to which the foam boards had been applied (also called a “Super-Chilled Echo U Container”) exhibited insulating performance at least several percentage points higher than that of the foamed polystyrene container.\",\n \"The thickness of the insulating material applied to the cardboard container to which the foam boards had been applied was 20 mm, whereas the thickness of the foamed polystyrene container was 25 mm, so if we consider materials of equal thickness, the results for the insulating material of the cardboard container to which the foam boards had been applied suggest that this foam has thermal insulation properties at least 25% better than those of standard foamed polystyrene.\",\n \"TABLE 4 Table of measurement results and calculated adiabatic coefficients Measurement test location Measurement site A Measurement site B Test apparatus Thermostatic tank Climatron Measured container* foamed foamed board polystyrene board polystyrene Ambient temperature 19.9° C. 19.8° C. 15.1° C. 15.6° C. Average temperature 51.0° C. 48.2° C. 49.3° C. 47.2° C. inside container Temperature 31.1° C. 28.4° C. 34.2° C. 31.6° C. differential Heater input power 9.53 W 9.60 W 9.65 W 9.67 W Fan input power 0.148 W 0.148 W 0.161 W 0.163 W Total heater unit 9.68 W 9.75 W 9.81 W 9.83 W input power Container adiabatic 0.311 W/° C. 0.343 W/° C. 0.278 W/° C. 0.311 W/° C. coefficient *Measured containers: “Foamed board” is short for a cardboard container lined with foamed boards, while “polystyrene” is short for a foamed polystyrene container used as a control.\",\n \"As seen in Table 4, the adiabatic coefficient of the cardboard container to which the foam boards had been applied was 0.311 W/° C. at measurement site A and 0.287 W/° C. at B, while the adiabatic coefficients of the control foamed polystyrene container were 0.343 W/° C. and 0.311 W/° C., respectively.\",\n \"The result is that the insulating performance of the cardboard container to which the foam boards had been applied was excellent (10.3% at measurement site A and 8.4% at B).\",\n \"However, the adiabatic coefficient varies by about 10% between the two measurement sites.\",\n \"The main reason for this seems to be the difference in the amount of air circulating inside the thermostatic apparatus keeping the ambient temperature of the container constant.\",\n \"In other words, at measurement site B the measurement was made in a relatively large climatron in which people can enter and work, whereas measurement site A was a much smaller thermostatic tank.\",\n \"The amount of air moving in a thermostatic tank generally tends to be large, and if the amount of air moving around an insulated box is larger, this is equivalent to the insulated container having a larger surface area, so the analysis was that the apparent insulation performance generally decreases.\",\n \"Therefore, a relative comparison under equal conditions is important in performing an evaluation such as this.\",\n \"This measurement and evaluation involved measuring the insulation performance of the same container at difference measurement test locations, and since the results for both tended to be the same, with an extremely small difference therebetween, and with the above-mentioned effect of the amount of air movement factored in, it was concluded that the data were a good match.\",\n \"In Example 2 above we discussed the manufacture of a foam board, but a foam sheet can also be manufactured by providing a roll molding machine comprising an upper roll and a lower roll to the rear of this manufacturing apparatus, and squeezing the above-mentioned foam board in its thickness direction by passing it through this roll molding machine.\",\n \"Industrial Applicability With the present invention, when a foam is manufactured using a starch as one polymer compound to be melted and mixing it with another polymer compound having a different melt viscosity, uniformity is ensured in the fluidity of the molten mixture, which is an essential requirement for stabilizing foam quality, and the resulting foam will have thin cell membranes and excellent thermal insulation performance, allowing a high-quality foam to be manufactured stably on an industrial scale.\",\n \"Also, this foam will be favorable in terms of protecting the global environment.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473881"}}},{"rowIdx":760,"cells":{"text":{"kind":"list like","value":[["Protein analysis","A method of forming an array of proteins selected from antigens or antibodies; said method comprising the steps of (i) expressing in a recombinant cell, a fusion protein which comprises either (a) an antigen or (b) an antibody binding protein, fused to a peptide having up to 50 amino acids, which peptide comprises amino acid sequence of SEQ ID NO 1 LX1X2IX3X4X5X6KX7X8X9X10 (SEQ ID NO 1) where X1 is a naturally occurring amino acid, X2 is any naturally occurring amino acid other than leucine, valine, isoleucine, tryptophan, phenylalanine or tyrosine, X3 is phenylalanine or leucine, X4 is glutamine or asparagine, X5 is alanine, glycine, serine or threonine, X6 is glycine or methionine, X7 is isoleucine, methionine or valine, X8 is glutamine, leucine, valine, tyrosine or isoleucine, X9 is tryptophan, tyrosine, valine, phenylalanine, leucine or isoleucine and X10 is any naturally occurring amino acid other than asparagine or glutamine; where said peptide is capable of being biotinylated by a biotin ligase at the lysine residue adjacent to X6; (ii) biotinylating said peptide of the fusion protein at the lysine residue adjacent X6; (iii) isolating the biotinylated fusion protein; (iv) applying the biotinylated fusion protein to an avidin or streptavidin coated non-porous support; (v) forming an array of at least three different proteins on the support by either (a) where the fusion protein comprises an antigen, carrying out steps (i) to (iv) the desired number of times to form an antigen array; or (b) where the fusion protein comprises an antibody binding protein, applying to said protein, either prior to or after step (iv) a plurality of different antibodies or binding fragments thereof."],["1.A method of forming an array of proteins selected from antigens or antibodies; said method comprising the steps of (i) expressing in a recombinant cell, a fusion protein which comprises either (a) an antigen or (b) an antibody binding protein, fused to a peptide having up to 50 amino acids, which peptide comprises an amino acid sequence of SEQ ID NO:1 LX1X2IX3X4X5X6KX7X8X9X10 (SEQ ID NO: 1) where X1 is a naturally occurring amino acid, X2 is any naturally occurring amino acid other than leucine, valine, isoleucine, tryptophan, phenylalanine or tyrosine, X3 is phenylalanine or leucine, X4 is glutamic acid or aspartic acid, X5 is alanine, glycine, serine or threonine, X6 is glutamine or methionine, X7 is isoleucine, methionine or valine, X8 is glutamic acid, leucine, valine, tyrosine or isoleucine, X9 is tryptophan, tyrosine, valine, phenylalanine, leucine and isoleucine and X10 is any naturally occurring amino acid other than aspartic acid or glutamic acid; where said peptide is capable of being biotinylated by a biotin ligase at the lysine residue adjacent to X6; (ii) biotinylating said peptide of the fusion protein at the lysine residue adjacent X6; (iii) isolating the biotinylated fusion protein; (iv) applying the biotinylated fusion protein to an avidin or streptavidin coated non-porous support; (v) forming an array of at least three different proteins on the support by either (a) where the fusion protein comprises an antigen, carrying out steps (i) to (iv) the desired number of times to form an antigen array; or (b) where the fusion protein comprises an antibody binding protein, applying to said protein, either prior to or after step (iv), a plurality of different antibodies or binding fragments thereof.","2.A method according to claim 1 wherein the peptide of SEQ ID NO:1 is selected from Leu His His Ile Leu Asp Ala Gln (SEQ ID NO; 30) Lys Met Val Trp Asn His Arg; Leu Asn Ala Ile Phe Glu Ala Met (SEQ ID NO: 33) Lys Met glu Tyr Ser Gly; Leu Gly Gly Ile Phe Glu Ala Met (SEQ ID NO: 34) Lys Met Glu Leu Arg Asp; Leu Ser Asp Ile Phe Glu Ala Met (SEQ ID NO: 37) Lys Met Val Tyr Arg Pro Cys; Leu Ser Asp Ile Phe Asp Ala Met (SEQ ID NO: 39) Lys Met Val Tyr Arg Pro Gln; Leu Lys Gly Ile Phe Glu Ala Met (SEQ ID NO: 41) Lys Met Glu Tyr Thr Ala Met; Leu Glu Gly Ile Phe Glu Ala Met (SEQ ID NO: 42) Lys Met Glu Tyr Ser Asn Ser; Leu Lys Glu Ile Phe Glu Gly Met (SEQ ID NO: 47) Lys Met Glu Phe Val Lys Pro; Arg Pro Val Leu Glu Asn Ile Phe (SEQ ID NO: 50) Glu Ala Met Lys Met Glu Val Trp Lys Pro; Thr Arg Ala Leu Leu Glu Ile Phe (SEQ ID NO: 57) Asp Ala Gln Lys Met Leu Tyr Gln His Leu; Met Ala Ser Ser Leu Arg Gln Ile (SEQ ID NO: 73) Leu Asp Ser Gln Lys Met Glu Trp Arg Ser Asn Ala Gly Gly Ser; Met Ala His Ser Leu Val Pro Ile (SEQ ID NO: 75) Phe Asp Ala Gln Lys Ile Glu Trp Art Asp Pro Phe Gly Gly Ser; Met Gly Pro Asp Leu Val Asn Ile (SEQ ID NO: 76) Phe Glu Ala Gln Lys Ile Glu Trp His Pro Leu Thr Gly Gly Ser; Met Ala Phe Ser Leu Arg Ser Ile (SEQ ID NO: 77) Leu Glu Ala Gln Lys Met Glu Leu Arg Asn Thr Pro Gly Gly Ser; Met Ala Gly Gly Leu Asn Asp Ile (SEQ ID NO: 78) Phe Glu Ala Gln Lys Ile Glu Trp His Glu Asp Thr Gly Gly Ser; Met Ser Ser Tyr Leu Ala Pro Ile (SEQ ID NO: 79) Phe Glu Ala Gln Lys Ile Glu Trp His Ser Ala Tyr Gly Gly Ser; Met Ala Lys Ala Leu Gln Lys Ile (SEQ ID NO: 80) Leu Glu Ala Gln Lys Met Glu Trp Arg Ser His Pro Gly Gly Ser; Met Ala Gly Ser Leu Ser Thr Ile (SEQ ID NO: 82) Phe Asp Ala Gln Lys Ile Glu Trp His Val Gly Lys Gly Gly Ser; Met Ala Gln Gln Leu Pro Asp Ile (SEQ ID NO: 83) Phe Asp Ala Gln Lys Ile Glu Trp Arg Ile Ala Gly Gly Gly Ser; Met Ala Gln Arg Leu Phe His Ile (SEQ ID NO: 84) Leu Asp Ala Gln Lys Ile Glu Trp His Gly Pro Lys Gly Gly Ser; Met Ala Gly Cys Leu Gly Pro Ile (SEQ ID NO: 85) Phe Glu Ala Gln Lys Met Glu Trp Arg His Phe Val Gly Gly Ser; Met Ala Trp Ser Leu Lys Pro Ile (SEQ ID NO: 86) Phe Asp Ala Gln Lys Ile Glu Trp His Ser Pro Gly Gly Gly Ser; Met Ala Leu Gly Leu Thr Arg Ile (SEQ ID NO: 87) Leu Asp Ala Gln Lys Ile Glu Trp His Arg Asp Ser Gly Gly Ser; and Met Ala gly Ser Leu Arg Gln Ile (SEQ ID NO: 88) Leu Asp Ala Gln Lys Ile Glu Trp Arg Arg Pro Leu Gly Gly Ser.","3.A method of forming an array of proteins selected from antigens or antibodies; said method comprising the steps of (i) expressing in a recombinant cell, a fusion protein which comprises either (a) an antigen or (b) an antibody binding protein, fused to a peptide having up to 50 amino acids or a fragment thereof having at least 13 amino acids, which peptide comprises the sequence selected from Leu Glu Glu Val Asp Ser Thr Ser (SEQ ID NO: 14) Ser Ala Ile Phe Asp Ala Met Lys Met Val Trp Ile Ser Pro Thr Glu Phe Arg; Gln Gly Asp Arg Asp Glu Thr Leu (SEQ ID NO: 15) Pro Met Ile Leu Arg Ala Met Lys Met Glu Val Tyr Asn Pro Gly Gly His Glu Lys; Ser Lys Cys Ser Tyr Ser His Asp (SEQ ID NO: 16) Leu Lys Ile Phe Glu Ala Gln Lys Met Leu Val His Ser Tyr Leu Arg Val Met Tyr Asn Tyr; Met Ala Ser Ser Asp Asp Gly Leu (SEQ ID NO: 17) Leu Thr Ile Phe Asp Ala Thr Lys Met Met Phe Ile Arg Thr; Ser Tyr Met Asp Arg Thr Asp Val (SEQ ID NO: 18) Pro Thr Ile Leu Glu Ala Met Lys Met Glu Leu His Thr Thr Pro Trp Ala Cys Arg; Ser Phe Pro Pro Ser Leu Pro Asp (SEQ ID NO: 19) Lys Asn Ile Phe Glu Ala Met Lys Met Tyr Val Ile Thr; Ser Val Val Pro Glu Pro Gly Trp (SEQ ID NO: 20) Asp Gly Pro Phe Glu Ser Met Lys Met Val Tyr His Ser Gly Ala Gln Ser Gly Gln; Val Arg His Leu Pro Pro Pro Leu (SEQ ID NO: 21) Pro Ala Leu Phe Asp Ala Met Lys Met Glu Phe Val Thr Ser Val Gln Phe; Asp Met Thr Met Pro Thr Gly Met (SEQ ID NO: 22) Thr Lys Ile Phe Glu Ala Met Lys Met Glu Val Ser Thr; Ala Thr Ala Gly Pro Leu His Glu (SEQ ID NO: 23) Pro Asp Ile Phe Leu Ala Met Lys Met Glu Val Val Asp Val Thr Asn Lys Ala Gly Gln; Ser Met Trp Glu Thr Leu Asn Ala (SEQ ID NO: 24) Gln Lys Thr Val Leu Leu; Ser His Pro Ser Gln Leu Met Thr (SEQ ID NO: 25) Asn Asp Ile Phe Glu Gly Met Lys Met Leu Tyr His; Thr Ser Glu Leu Ser Lys Leu Asp (SEQ ID NO: 27) Ala Thr Ile Phe Ala Ala Met Lys Met Gln Trp Trp Asn Pro Gly; Val Met Glu Thr Gly Leu Asp Leu (SEQ ID NO: 28) Arg Pro Ile Leu Thr Gly Met Lys Met Asp Trp Ile Pro Lys; Pro Gln Gly Ile Phe Glu Ala Gln (SEQ ID NO: 31) Lys Met Leu Trp Arg Ser; Leu Ala Gly Thr Phe Glu Ala Leu (SEQ ID NO: 32) Lys Met Ala Trp His Glu His; Leu Leu Arg Thr Phe Glu Ala Met (SEQ ID NO: 35) Lys Met Asp Trp Arg Asn Gly; Leu Ser Thr Ile Met Glu Gly Met (SEQ ID NO: 36) Lys Met Tyr Ile Gln Arg Ser; Leu Glu Ser Met Leu Glu Ala Met (SEQ ID NO: 38) Lys Met Gln Trp Asn Pro Gln; Leu Ala Pro Phe Phe Glu Ser Met (SEQ ID NO: 40) Lys Met Val Trp Arg Glu His; Leu Leu Gln Thr Phe Asp Ala Met (SEQ ID NO: 43) Lys Met Glu Trp Leu Pro Lys; Val Phe Asp Ile Leu Glu Ala Gln (SEQ ID NO: 44) Lys Val Val Thr Leu Arg Phe; Leu Val Ser Met Phe Asp Gly Met (SEQ ID NO: 45) Lys Met Glu Trp Lys Thr Leu; Leu Glu Pro Ile Phe Glu Ala Met (SEQ ID NO: 46) Lys Met Asp Trp Arg Leu Glu; Leu Gly Gly Ile Glu Ala Gln Lys (SEQ ID NO: 48) Met Leu Leu Tyr Arg Gly Asn; Arg Ser Pro Ile Ala Glu Ile Phe (SEQ ID NO: 51) Glu Ala Met Lys Met Glu Tyr Arg Glu Thr; Gln Asp Ser Ile Met Pro Ile Phe (SEQ ID NO: 52) Glu Ala Met Lys Met Ser Trp His Val Asn; Asp Gly Val Leu Phe Pro Ile Phe (SEQ ID NO: 53) Glu Ala Met Lys Met Ile Arg Leu Glu Thr; Val Ser Arg Thr Met Thr Asn Phe (SEQ ID NO: 54) Glu Ala Met Lys Met Ile Tyr His Asp Leu; Asp Val Leu Leu Pro Thr Val Phe (SEQ ID NO: 55) Glu Ala Met Lys Met Tyr Ile Thr Lys; Pro Asn Asp Leu Glu Arg Ile Phe (SEQ ID NO: 56) Asp Ala Met Lys Ile Val Thr Val His Ser; Arg Asp Val His Val Gly Ile Phe (SEQ ID NO: 58) Glu Ala Met Lys Met Tyr Thr Val Glu Thr; Gly Asp Lys Leu Thr Glu Ile Phe (SEQ ID NO: 59) Glu Ala Met Lys Ile Gln Trp Thr Ser Gly; Leu Glu Gly Leu Arg Ala Val Phe (SEQ ID NO: 60) Glu Ser Met Lys Met Glu Leu Ala Asp Glu; Val Ala Asp Ser His Asp Thr Phe (SEQ ID NO: 61) Ala Ala Met Lys Met Val Trp Leu Asp Thr; Gly Leu Pro Leu Gln Asp Ile Leu (SEQ ID NO: 62) Glu Ser Met Lys Ile Val Met Thr Ser Gly; Arg Val Pro Leu Glu Ala Ile Phe (SEQ ID NO: 63) Glu Gly Ala Lys Met Ile Trp Val Pro Asn Asn; Pro Met Ile Ser His Lys Asn Phe (SEQ ID NO: 64) Glu Ala Met lys Met Lys Phe Val Pro Glu; Lys Leu Gly Leu Pro Ala Met Phe (SEQ ID NO: 65) Glu Ala Met Lys Met Glu Trp His Pro Ser; Gln Pro Ser Leu Leu Ser Ile Phe (SEQ ID NO: 66) Glu Ala Met Lys Met Gln Ala Ser Leu Met; Leu Leu Glu Leu Arg Ser Asn Phe (SEQ ID NO: 67) Glu Ala Met Lys Met Glu Trp Gln Ile Ser; Asp Glu Glu Leu Asn Gln Ile Phe (SEQ ID NO: 68) Glu Ala Met Lys Met Tyr Pro Leu Val His Val Thr Lys; Ser Asn Leu Val Ser Leu Leu His (SEQ ID NO: 70) Ser Gln Lys Ile Leu Trp Thr Asp Pro Gln Ser Phe Gly; Leu Phe Leu His Asp Phe Leu Asn (SEQ ID NO: 71) Ala Gln Lys Val Glu Leu Try Pro Val Thr Ser Ser Gly; Ser Asp Ile Asn Ala Leu Leu Ser (SEQ ID NO: 72) Thr Gln Lys Ile Tyr Trp Ala His; Met Ala Phe Gln Leu Cys Lys Ile (SEQ ID NO: 81) Phe Try Ala Gln Lys Met Clu Trp His Gly Val Gly Gly Gly Ser, and; Met Ala Asp Arg Leu Ala Tyr Ile (SEQ ID NO: 89) Leu Glu Ala Gln Lys Met Glu Trp His Pro His Lys Gly Gly Ser, where said peptide is capable of being biotinylated by a biotin ligase; (ii) biotinylating said peptide of the fusion protein; (iii) isolating the biotinylated fusion protein; (iv) applying the biotinylated fusion protein to an avidin or streptavidin coated non-porous support; (v) forming an array of at least three different proteins on the support by either (a) where the fusion protein comprises an antigen, carrying out steps (i) to (iv) the desired number of times to form an antigen array; or (b) where the fusion protein comprises an antibody binding protein, applying to said protein, either prior to or after step (iv), a plurality of different antibodies or binding fragments thereof.","4.A method according to claim 1 wherein the fusion protein further comprises a second peptide sequence capable of acting as an affinity or detection tag sequence to the fusion protein wherein the sequence comprises between 1 and 30 amino acids.","5.A method according to claim 4 wherein the second peptide sequence is fused to the end of the amino acid sequence of SEQ ID NO:1 or a peptide of 13-50 amino acids comprising a sequence selected from SEQ ID NOS: 14-25, 27, 28, 31, 32, 35, 36, 38, 40, 43-46, 48, 51-56, 58-68, 70-72, 81 and 89.6.A method according to claim 4 wherein the second peptide sequence is fused to the opposite end of the antigen or antibody binding protein to which the amino acid sequence of SEQ ID NO: 1 or a peptide of 13-50 amino acids comprising a sequence selected from SEQ ID NOS: 14-25, 27, 28, 31, 32, 35, 36, 38, 40, 43-46, 48, 51-56, 58-68, 70-72, 81 and 89 is fused.","7.A method according to claim 4 wherein at least one amino acid of the peptide sequence tag is histidine.","8.A method according to claim 7 wherein the peptide sequence tag has the formula His-X in which X is selected from -Gly-, -His-, -Tyr-, -Gly-, -Trp-, -Val-, -Leu-, -Ser-, -Lys-, -Phe-, -Met-, -Ala-, -Glu-, -Ile-, -Thr-, -Asp-, -Asn-, -Gln-, -Arg-, -Cys- and -Pro-.","9.A method according to claim 7 wherein the peptide sequence tag has the formula Y-His.","10.A method according to claim 9 wherein Y is selected from -Gly-, -Ala-, -His-, and -Tyr-.","11.A method according to claim 1 wherein the recombinant cell expresses biotin ligase and step (ii) is effected in the presence of biotin such that biotinylation occurs in vivo in said cell.","12.A method according to claim 11 wherein recombinant cell expresses biotin.","13.A method according to claim 1 wherein step (iii) is effected using a further antibody or a binding fragment thereof, which is specific for the peptide of SEQ ID NO:1 or a peptide of 13-50 amino acids comprising a sequence selected from SEQ ID NOS: 14-25, 27, 28, 31, 32, 35, 36, 38, 40, 43-46, 48, 51-56, 58-68, 70-72, 81 and 89.14.A method according claim 4 wherein step (iii) is effected using a further antibody or a binding fragment thereof, which is specific for the said second peptide sequence.","15.A method according to claim 13 wherein said further antibody or binding fragment thereof is immobilised on a column, magnetic bead or loaded into a pipette tip.","16.A method according to claim 15 wherein bound fusion protein is subsequently eluted by increasing the pH conditions.","17.A method according to claim claim 1 wherein in step (iii) the fusion protein is isolated using a separation material which releasably binds biotin.","18.A method according to claim 17 wherein the separation material is a modified version of avidin or streptavidin, which has lower affinity for biotin than native avidin or streptavidin.","19.A method according to claim 17 wherein the separation material is attached to magnetic beads or pipette tips.","20.A method according to claim 17 wherein the fusion protein is eluted from the separation material by changing the pH conditions.","21.A method according to claim 1 wherein some areas of the coated support used in step (iv) are blocked to prevent binding of the fusion protein thereto.","22.A method according to claim 1 wherein the peptide is a peptide of 15 amino acids in length.","23.A method according to claim 22 wherein the peptide is of SEQ ID NO:2 Gly Leu Asn Asp lie Phe Glu Ala Gln Lys Ile Glu Trp His Glu (SEQ ID NO:2).","24.A method according to any one of tho preceding claims claim 1 wherein the fusion protein comprises an antigen.","25.A method according to claim 24 wherein an antigen library is used to create the array.","26.A method according to claim 1 wherein the fusion protein comprises an antibody binding protein.","27.A method according to claim 26 wherein the antibody binding protein is one or more of Protein A, Protein G and Protein L. 28.A method according to claim 23 wherein the antibody binding protein comprises a mixture of Protein A, Protein G and Protein L. 29.A method according to claim 26 wherein the antibody binding protein may be fused to the said peptide at the N-terminus thereof or it may be fused to said peptide at the C-terminus thereof.","30.A method according to claim 1 wherein prior to step (iv), the identity of the expressed fusion protein is confirmed.","31.A method according to claim 30 wherein the identity is confirmed using mass spectrometry.","32.A method according to claim 1 wherein protein normalisation is carried out by detecting the peptide of SEQ ID NO:1 or a peptide of 13-50 amino acids comprising a sequence selected from SEQ ID NOS: 14-25, 27, 28, 31, 32, 35, 36, 38.40, 43-46, 48, 51-56, 58-68, 70-72, 81 and 89 in the fusion protein which acts as an internal control.","33.A method according to claim 1 wherein protein normalisation is carried out by detecting the peptide sequence tag comprising between 1 and 30 amino acids in the fusion protein which acts as an internal control.","34.A method according to claim 32 wherein the peptide is detected by an antibody with a high affinity for the said peptide.","35.A method according to claim 32 wherein the protein normalisation is effected by performing an immunoassay simultaneously with subsequent analysis of a biological sample using the array.","36.A method according to claim 1 wherein the avidin or streptavidin coated non-porous support used in step (iv) is a glass or plastics material.","37.A method according to claim 1 wherein a further acceptor layer is provided on top of the foundation of the streptavidin layer on the support.","38.A method according to claim 1 wherein the array comprises from 3-10,000 different fusion proteins.","39.A method according to claim 38 wherein each protein is present in a form in which the peptide including SEQ ID NO:1 or a peptide of 13-50 amino acids comprising a sequence selected from SEQ ID NOS: 14-25, 27, 28, 31, 32, 35.36.38, 40, 43-46, 48, 51-56, 58-68, 70-72, 81 and 89 is fused to the C-terminus, and also in a form in which the peptide including SEQ ID NO:1 or a peptide of 13-50 amino acids comprising a sequence selected from SEQ ID NOS: 14-25, 27, 28, 31, 32, 35, 36, 38, 40, 43-46.48, 51-56, 58-68, 70-72, 81 and 89 is fused to the N-terminus.","40.A protein array obtained by a method according to claim 1.41.A method of detecting binding between an antibody and an antigen, said method comprising the steps of: (vi) applying to the array according to claim 40 a sample which contains or is suspected of containing an antibody in the case of an array of step (v)(a), or an antigen in the case of the array of step (v) (b); and (vii) detecting bound antibody or antigen on the support.","42.A method according to claim 41 wherein step (vii) is carried out by ELISA methods.","43.A method according to claim 41 wherein the fusion protein array continues to be monitored for quality and/or the density of the protein during step (vi) and/or step (vii).","44.A method according to claim 43 wherein the monitoring is effected by detecting the peptide which comprises SEQ ID NO:1 or a peptide of 13-50 amino acids comprising a sequence selected from SEQ ID NOS: 14-25, 27, 28, 31, 32, 35, 36, 38, 40, 43-46, 48, 51-56, 58-68, 70-72, 81 and 89.45.A method according to claim 43 wherein array comprises fusion proteins which further comprise a second peptide sequence, and monitoring is effected by detecting the presence of the second peptide sequence, wherein the second peptide sequence comprises between 1 and 30 amino acids.","46.A method according to claim 1 wherein at least some of the steps are operated automatically.","47.A method according to claim 46 wherein all the steps of the method are operated automatically.","48.A fusion protein comprising an antibody binding protein fused at the N— or C-terminus to a peptide of 13 to 50 amino acids, which comprises SEQ ID NO:1 or a peptide of 13-50 amino acids comprising a sequence selected from SEQ ID NOS: 14-25, 27, 28, 31, 32, 35, 36, 38, 40, 43-46, 48, 51-56, 58-68, 70-72, 81 and 89.49.A fusion protein according to claim 48 wherein the peptide is a peptide of SEQ ID NO:2.50.A fusion protein according to claim 48 further comprising a second peptide sequence which acts as a tag sequence to the fusion protein wherein the sequence comprises between 1 and 20 amino acids.","51.A fusion protein according to claim 48 wherein the antibody binding protein is Protein A, G or L or a mixture thereof.","52.A nucleic acid sequence, which encodes the fusion protein according to claim 48.53.A nucleic acid according to claim 52 wherein the sequence which encodes the peptide is of SEQ ID NO:9 GGCCTGAACGACATCTTCGAGGCTCAGAAAATCGAATGGCACGAA (SEQ ID NO:9)."],["<SOH> BACKGROUND <EOH>The concept of attaching a number of different proteins to surface supports to form an “array” of proteins has been widely described in the literature (see for example EP0063810, WO84/03151, U.S. Pat.","No.","5,143,854).","Recently, there has been a growing interest in the concept of manufacturing devices whereby large numbers of proteins of various classes are arrayed onto different types of solid supports.","Examples include antigen, antibody, protein (protein-protein interaction) and functional enzymes arrays.","The background to the technology, and the potential uses for such devices, are thoroughly catalogued in the literature (Joos et al Electrophoresis 2000, 21, 2641-2650, Haab et al Genome Biology 2000 1(6), Borrebaeck Immunology Today, August 2000) and examples of potential utility can be found in a number of recent patent applications including WO 00/07024, WO 99/40434, WO 99/39210 and WO00/54046.The concept of creating antigen arrays was described in EP 0063810 in 1982.It was reported that antigens and antibodies could be bound to a porous solid support enabling an unlimited number of antibody-antigen interactions to be conducted simultaneously.","To make antigen arrays, antigens were simply aliquoted in very small volumes onto nitrocellulose membranes or similar supports, allowed to adsorb and then probed with the corresponding antibodies.","As with Enzyme Linked Immunosorbent Assay (ELISA) protocols performed in solution or in plastic plates, non-specific interactions were blocked with Bovine Serum Albumin (BSA), and this is now standard practise.","It was also reported that the elements (or spots) of the array did not diffuse and were adsorbed tightly onto the membrane.","It should be noted, however, that the dimensions for these elements were considerably larger than those obtained in micro array device systems.","EP 0063810 describes how the protein arrays could be made by aliquoting proteins by hand, using mechanical procedures including a “charged drop” or lithographic process.","In this manner elements with a diameter of less than 500 microns (compared with 100 microns that can be achieved with current automated array systems) were produced.","However, one of the main disadvantages associated with the use of membranes as opposed to non-porous surfaces is that the elements tend to diffuse through the support material unless there is immediate binding.","Attempts were made to overcome this problem.","U.S. Pat.","No.","4,496,654 describes use of porous surfaces such as paper disks which were treated with streptavidin (which is adsorbed onto the surface) enabling arrays of biotinylated antibodies to be arranged in any desired pattern.","Following blocking with BSA, the paper discs could be probed with the antigen (exemplified with human chorionic gonadotropin) which could then be detected with an enzyme assay.","The biotinylated antibody immediately bound very tightly to the surface of the paper reducing diffusion of the spots.","To achieve this using an “acceptor” surface such as an avidin or streptavidin coated surface, requires that each antibody and antigen, which is attached to the array, must be biotinylated prior to attachment to the array with no guarantee that this process will not impair its avidity (or antigenicity if an antigen is used) compared with the native protein.","Non-porous surfaces also have the disadvantage that they are not as robust as solid surfaces, including various types of glass or plastics, and so cannot be washed or treated as stringently.","For antigen and antibody arrays, it has been found however that attaching a protein to a solid surface generally leads to a reduction in antigenicity of the antigen and avidity of the antibody compared with that observed when the antigen or antibody is in free solution.","Previous attempts (see WO84/03 151 and Haab et al 2000 supra.)","to immobilise antigens and antibodies were not greatly successful.","WO84/03 151 describes that antibodies can be applied directly onto glass surfaces such as a microscope cover slip and dried.","When blocked and then exposed to antigens, in this case in the form of whole cells, the antigens were captured by the array.","However, WO84/03 151 further describes that these antigens needed to be added at a higher concentration compared with the equivalent ELISA performed in solution.","It was also noted that the antibodies had to be “highly enriched in order to achieve a sufficiently dense antibody coat for the desired cell adherence”.","It also took considerable time for the antibodies to be adsorbed onto the glass surface.","Other approaches for the direct immobilisation of antigens and antibodies have been reported.","One approach was to first adsorb calcium phosphate in the form of hydroxyapatite onto filter papers onto which proteins were bound by ionic interaction as described in U.S. Pat.","No.","5,827,669.These inventors reported that this was not effective for acidic proteins and that the antibodies suffered from “bad orientation” onto the Ca/phosphate layer.","Success was, however, reported when this method was used in immobilising streptavidin.","Another method for immobilising proteins to solid, non-porous surfaces included attaching them using an adhesive polyphenolic protein isolated from muscles as described in U.S. Pat.","No.","5,817,470.By coating solid surfaces, such as a polystyrene multi-well plate with polyphenolic protein, various antigens could be bound to the treated support and detected in an ELISA sandwich comprising of a primary antibody followed by a secondary antibody conjugated to an enzyme.","However, the inventors conceded that the procedure was limited by the amount of antigen bound or adsorbed to the solid surface.","The final amount of antigen strongly bound to the surface of the plate varied depending on a number of factors such as the molecular characteristics of the antigens, the properties of the solid support, the concentration of the antigen in the solution as well as the characteristics of the buffer used to dissolve the antigen used to coat or to activate the surface.","In general, only a small fraction of the antigen present in the coating solution was adsorbed to the surface.","Direct attachment of antibodies and antigens to non-porous surfaces was also been attempted with a collection of 113 antibodies and their corresponding antigens (Haab et al, 2000 Genome Biology 1(6)).","By exploiting technology developed for DNA microarrays, glass slides coated with poly-s-lysine were used to immobilise both antigens in one experiment and antibodies in another.","The results reported showed that only 50% of the arrayed antigens and 20% of the arrayed antibodies, provided specific and accurate measurements of their cognate ligands at or below concentrations of 1.60 μl/ml and 0.34 μg/ml respectively.","The high failure rate in binding antibodies to solid surfaces would not be acceptable for a large-scale antibody array manufacturing programme.","This supports the view that direct attachment of antigens and antibodies is an unsuitable technique to retain antibody/antigen functionality if protein arrays are to fulfil their potential.","The use of coatings such as avidin and streptavidin as binders for biotin labelled proteins is well known for use in conjunction with many proteins.","The proteins are generally isolated first, and then biotinylated.","Biotin can be conjugated to the protein at any or all active lysine sites contained within it.","Thus, when antigens or antibodies are biotinylated in this way, biotin groups may be present at their N-terminal groups and at any number of potential active lysine residues over their surface.","This means that they will adopt any number of different orientations once bound to the streptavidin layer and so the binding properties will be diverse.","Furthermore, access to the antigen or antibody immobilised via streptavidin will be reduced by steric hindrance, leading to generally inadequate assay.","It has been found that it is possible to reduce the steric hindrance and increase the sensitivity of the immunoassay by including a linker between the antigen/antibody and the biotinylated site.","U.S. Pat.","No.","5,811,246 describes how small synthetic peptides used in either immunoassays or for raising antisera can be linked to a “carrier” protein such as avidin or streptavidin via a linker such as various bradykinin derivatives.","This has several advantages.","Firstly, the condensation reaction between the free N-terminal group on the peptide and the linker preserves the charged residues essential for recognition by an antibody (immunoassay) or to elicit an immune response (immunisation).","Secondly, the bradykinin linker can then be biotinylated in such a way as to preserve the free charged groups on the small peptide.","In each case, the presence of the linker appears to promote a more sensitive immunoassay and an improved immune response when used as an immunising agent.","This use of a bradykinin derivative in this way however introduces further steps and complications into the process.","Biotinylated peptides fused to peptides or proteins of interest are described in U.S. Pat.","Nos.","5,723,584, 5,874,239 and 5,932,433, and further in Beckett et a!.","Protein Sci.","(1999) 8(4) 921-9.These peptides are used in order to biotinylate recombinant proteins, so as to allow rapid purification, immobilization, labelling and detection thereof.","It is not suggested that these peptides should be used in particular with antigens or antibody binding proteins, or that they should be formulated in arrays.","The present applicants have found that the peptides used in these patents allow the production of very good antigen or antibody arrays, which can be efficiently produced on non-porous supports whilst substantially retaining the binding avidity of these proteins."],["<SOH> SUMMARY OF THE INVENTION <EOH>According to a first aspect of the present invention there is provided a method of forming an array of proteins selected from antigens or antibodies; said method comprising the steps of (i) expressing in a recombinant cell, a fusion protein which comprises either (a) an antigen or (b) an antibody binding protein, fused to a peptide having up to 50 amino acids, which peptide comprised amino acid sequence of SEQ ID NO 1 LX 1 X 2 IX 3 X 4 X 5 X 6 KX 7 X 8 X 9 X 10 (SEQ ID NO 1) where X1 is a naturally occurring amino acid, X 2 is any naturally occurring amino acid other than leucine, valine, isoleucine, tryptophan, phenylalanine or tyrosine, X 3 is phenylalanine or leucine, X 4 is glutamine or asparagine, X 5 is alanine, glycine, serine or threonine, X 6 is glycine or methionine, X 7 is isoleucine, methionine or valine, X 8 is glutamine, leucine, valine, tyrosine or isoleucine, X 9 is tryptophan, tyrosine, valine, phenylalanine, leucine or isoleucine and X 10 is any naturally occurring amino acid other than asparagine or glutamine; where said peptide is capable of being biotinylated by a biotin ligase at the lysine residue adjacent to X 6 ; (ii) biotinylating said peptide of the fusion protein at the lysine residue adjacent X 6 ; (iii) isolating the biotinylated fusion protein; (iv) applying the biotinylated fusion protein to an avidin or streptavidin coated non-porous support; (v) forming an array of at least three different proteins on the support by either (a) where the fusion protein comprises an antigen, carrying out steps (i) to (iv) the desired number of times to form an antigen array; or (b) where the fusion protein comprises an antibody binding protein, applying to said protein, either prior to or after step (iv) a plurality of different antibodies or binding fragments thereof.","The applicants have found that by using a fusion of the antigen or antibody binding protein to a peptide of SEQ ID NO 1, these proteins may be immobilised onto solid surfaces, whilst substantially maintaining the antigenicity of proteins, or the binding capabilities of the antibody binding proteins.","This may be because the fusion peptide is biotinylated rather than the protein itself, and so there is less disruption of the protein's antigenicity when attached to the support surface.","In addition, the peptide including SEQ ID NO 1 appears to reduce steric hindrance to enable interaction between antigen and antibody.","By ensuring that the peptide linker is attached at a terminal region of the protein, and contains the biotinylation site, sites on the protein which are essential for function appear to be largely unaffected.","This combination is particularly advantageous in the context of methods of analysis using antigens or antibody arrays.","The method described herein represents the first time that the mode of attachment of proteins to non-porous surfaces (step vi), the mode of protein isolation from cell lysate (step iii) and the method of biotinylation (step ii) utilise the same fusion peptide.","Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.","As used herein the expression “antibody binding protein” refers to proteins which are known to bind to regions of antibodies, or to mixtures of these.","Examples of such proteins include Protein A, Protein L and Protein G Antibody binding proteins are used in accordance with the invention in the production of antibody arrays.","The antibodies are bound by antibody binding proteins, such as Proteins A, G and/or L or a mixture of one or more of these, which are themselves anchored via the linker to the streptavidin coating on the support surface.","While biotinylated versions of native Protein A, G and L are commercially available and can be attached to the streptavidin coating on the support surface, the applicants have found that by fusing these proteins to biotinylated tags in accordance with the present invention at the C and/or N-terminals, highly effective binding of antibodies of various types was achieved.","This may also be the result of reduced steric factors, or that the binding sites on the proteins are all readily available.","In addition, by using the method of the invention, the biotinylated fusion protein is immediately captured on application to the avidin or streptavidin coated support in step (iv) leading to very discrete spots of protein on the support, with minimal observable diffusion.","Particular examples of peptides having up to 50 amino acids, which peptide comprises an amino acid sequence of SEQ ID NO 1 are listed in U.S. Pat.","No.","5,723,584, U.S. Pat.","No.","5,874,239 and U.S. Pat.","No.","5,932,433, the content of which are incorporated herein by reference.","Examples of peptides provided in these references are listed below: Leu Glu Glu Val Asp Ser Thr Ser (SEQ ID NO: 14) Ser Ala Ile Phe Asp Ala Met Lys Met Val Trp Ile Ser Pro Thr Glu Phe Arg; Gln Gly Asp Arg Asp Glu Thr Leu (SEQ ID NO: 15) Pro Met Ile Leu Arg Ala Met Lys Met Glu Val Tyr Asn Pro Gly Gly His Glu Lys; Ser Lys Cys Ser Tyr Ser His Asp (SEQ ID NO: 16) Leu Lys Ile Phe Glu Ala Gln Lys Met Leu Val His Ser Tyr Leu Arg Val Met Tyr Asn Tyr; Met Ala Ser Ser Asp Asp Gly Leu (SEQ ID NO: 17) Leu Thr Ile Phe Asp Ala Thr Lys Met Met Phe Ile Arg Thr; Ser Tyr Met Asp Arg Thr Asp Val (SEQ ID NO: 18) Pro Thr Ile Leu Glu Ala Met Lys Met Glu Leu His Thr Thr Pro Trp Ala Cys Arg; Ser Phe Pro Pro Ser Leu Pro Asp (SEQ ID NO: 19) Lys Asn Ile Phe Glu Ala Met Lys Met Tyr Val Ile Thr; Ser Val Val Pro Glu Pro Gly Trp (SEQ ID NO: 20) Asp Gly Pro Phe Glu Ser Met Lys Met Val Tyr His Ser Gly Ala Gln Ser Gly Gln; Val Arg His Leu Pro Pro Pro Leu (SEQ ID NO: 21) Pro Ala Leu Phe Asp Ala Met Lys Met Glu Phe Val Thr Ser Val Gln Phe; Asp Met Thr Met Pro Thr Gly Met (SEQ ID NO: 22) Thr Lys Ile Phe Glu Ala Met Lys Met Glu Val Ser Thr; Ala Thr Ala Gly Pro Leu His Glu (SEQ ID NO: 23) Pro Asp Ile Phe Leu Ala Met Lys Met Glu Val Val Asp Val Thr Asn Lys Ala Gly Gln; Ser Met Trp Glu Thr Leu Asn Ala (SEQ ID NO: 24) Gln Lys Thr Val Leu Leu; Ser His Pro Ser Gln Leu Met Thr (SEQ ID NO: 25) Asn Asp Ile Phe Glu Gly Met Lys Met Leu Tyr His; Thr Ser Glu Leu Ser Lys Leu Asp (SEQ ID NO: 27) Ala Thr Ile Phe Ala Ala Met Lys Met Gln Trp Trp Asn Pro Gly; Val Met Glu Thr Gly Leu Asp Leu (SEQ ID NO: 28) Arg Pro Ile Leu Thr Gly Met Lys Met Asp Trp Ile Pro Lys; Leu His His Ile Leu Asp Ala Gln (SEQ ID NO: 30) Lys Met Val Trp Asn His Arg; Pro Gln Gly Ile Phe Glu Ala Gln (SEQ ID NO: 31) Lys Met Leu Trp Arg Ser; Leu Ala Gly Thr Phe Glu Ala Leu (SEQ ID NO: 32) Lys Met Ala Trp His Glu His; Leu Asn Ala Ile Phe Glu Ala Met (SEQ ID NO: 33) Lys Met Glu Tyr Ser Gly; Leu Gly Gly Ile Phe Glu Ala Met (SEQ ID NO: 34) Lys Met Glu Leu Arg Asp; Leu Leu Arg Thr Phe Glu Ala Met (SEQ ID NO: 35) Lys Met Asp Trp Arg Asn Gly; Leu Ser Thr Ile Met Glu Gly Met (SEQ ID NO: 36) Lys Met Tyr Ile Gln Arg Ser; Leu Ser Asp Ile Phe Glu Ala Met (SEQ ID NO: 37) Lys Met Val Tyr Arg Pro Cys; Leu Glu Ser Met Leu Glu Ala Met (SEQ ID NO: 38) Lys Met Gln Trp Asn Pro Gln; Leu Ser Asp Ile Phe Asp Ala Met (SEQ ID NO: 39) Lys Met Val Tyr Arg Pro Gln; Leu Ala Pro Phe Phe Glu Ser Met (SEQ ID NO: 40) Lys Met Val Trp Arg Glu His; Leu Lys Gly Ile Phe Glu Ala Met (SEQ ID NO: 41) Lys Met Glu Tyr Thr Ala Met; Leu Glu Gly Ile Phe Glu Ala Met (SEQ ID NO: 42) Lys Met Glu Tyr Ser Asn Ser; Leu Leu Gln Thr Phe Asp Ala Met (SEQ ID NO: 43) Lys Met Glu Trp Leu Pro Lys; Val Phe Asp Ile Leu Glu Ala Gln (SEQ ID NO: 44) Lys Val Val Thr Leu Arg Phe; Leu Val Ser Met Phe Asp Gly Met (SEQ ID NO: 45) Lys Met Glu Trp Lys Thr Leu; Leu Glu Pro Ile Phe Glu Ala Met (SEQ ID NO: 46) Lys Met Asp Trp Arg Leu Glu; Leu Lys Glu Ile Phe Glu Gly Met (SEQ ID NO: 47) Lys Met Glu Phe Val Lys Pro; Leu Gly Gly Ile Glu Ala Gln Lys (SEQ ID NO: 48) Met Leu Leu Tyr Arg Gly Asn; Arg Pro Val Leu Glu Asn Ile Phe (SEQ ID NO: 50) Glu Ala Met Lys Met Glu Val Trp Lys Pro; Arg Ser Pro Ile Ala Glu Ile Phe (SEQ ID NO: 51) Glu Ala Met Lys Met Glu Tyr Arg Glu Thr; Gln Asp Ser Ile Met Pro Ile Phe (SEQ ID NO: 52) Glu Ala Met Lys Met Ser Trp His Val Asn; Asp Gly Val Leu Phe Pro Ile Phe (SEQ ID NO: 53) Glu Ala Met Lys Met Ile Arg Leu Glu Thr; Val Ser Arg Thr Met Thr Asn Phe (SEQ ID NO: 54) Glu Ala Met Lys Met Ile Tyr His Asp Leu; Asp Val Leu Leu Pro Thr Val Phe (SEQ ID NO: 55) Glu Ala Met Lys Met Tyr Ile Thr Lys; Pro Asn Asp Leu Glu Arg Ile Phe (SEQ ID NO: 56) Asp Ala Met Lys Ile Val Thr Val His Ser; Thr Arg Ala Leu Leu Glu Ile Phe (SEQ ID NO: 57) Asp Ala Gln Lys Met Leu Tyr Gln His Leu; Arg Asp Val His Val Gly Ile Phe (SEQ ID NO: 58) Glu Ala Met Lys Met Tyr Thr Val Glu Thr; Gly Asp Lys Leu Thr Glu Ile Phe (SEQ ID NO: 59) Glu Ala Met Lys Ile Gln Trp Thr Ser Gly; Leu Glu Gly Leu Arg Ala Val Phe (SEQ ID NO: 60) Glu Ser Met Lys Met Glu Leu Ala Asp Glu; Val Ala Asp Ser His Asp Thr Phe (SEQ ID NO: 61) Ala Ala Met Lys Met Val Trp Leu Asp Thr; Gly Leu Pro Leu Gln Asp Ile Leu (SEQ ID NO: 62) Glu Ser Met Lys Ile Val Met Thr Ser Gly; Arg Val Pro Leu Glu Ala Ile Phe (SEQ ID NO: 63) Glu Gly Ala Lys Met Ile Trp Val Pro Asn Asn; Pro Met Ile Ser His Lys Asn Phe (SEQ ID NO: 64) Glu Ala Met Lys Met Lys Phe Val Pro Glu; Lys Leu Gly Leu Pro Ala Met Phe (SEQ ID NO: 65) Glu Ala Met Lys Met Glu Trp His Pro Ser; Gln Pro Ser Leu Leu Ser Ile Phe (SEQ ID NO: 66) Glu Ala Met Lys Met Gln Ala Ser Leu Met; Leu Leu Glu Leu Arg Ser Asn Phe (SEQ ID NO: 67) Glu Ala Met Lys Met Glu Trp Gln Ile Ser; Asp Glu Glu Leu Asn Gln Ile Phe (SEQ ID NO: 68) Glu Ala Met Lys Met Tyr Pro Leu Val His Val Thr Lys; Ser Asn Leu Val Ser Leu Leu His (SEQ ID NO: 70) Ser Gln Lys Ile Leu Trp Thr Asp Pro Gln Ser Phe Gly; Leu Phe Leu His Asp Phe Leu Asn (SEQ ID NO: 71) Ala Gln Lys Val Glu Leu Tyr Pro Val Thr Ser Ser Gly; Ser Asp Ile Asn Ala Leu Leu Ser (SEQ ID NO: 72) Thr Gln Lys Ile Tyr Trp Ala His; Met Ala Ser Ser Leu Arg Gln Ile (SEQ ID NO: 73) Leu Asp Ser Gln Lys Met Glu Trp Arg Ser Asn Ala Gly Gly Ser; Met Ala His Ser Leu Val Pro Ile (SEQ ID NO: 75) Phe Asp Ala Gln Lys Ile Glu Trp Arg Asp Pro Phe Gly Gly Ser; Met Gly Pro Asp Leu Val Asn Ile (SEQ ID NO: 76) Phe Glu Ala Gln Lys Ile Glu Trp His Pro Leu Thr Gly Gly Ser; Met Ala Phe Ser Leu Arg Ser Ile (SEQ ID NO: 77) Leu Glu Ala Gln Lys Met Glu Leu Arg Asn Thr Pro Gly Gly Ser; Met Ala Gly Gly Leu Asn Asp Ile (SEQ ID NO: 78) Phe Glu Ala Gln Lys Ile Glu Trp His Glu Asp Thr Gly Gly Ser; Met Ser Ser Tyr Leu Ala Pro Ile (SEQ ID NO: 79) Phe Glu Ala Gln Lys Ile Glu Trp His Ser Ala Tyr Gly Gly Ser; Met Ala Lys Ala Leu Gln Lys Ile (SEQ ID NO: 80) Leu Glu Ala Gln Lys Met Glu Trp Arg Ser His Pro Gly Gly Ser; Met Ala Phe Gln Leu Cys Lys Ile (SEQ ID NO: 81) Phe Tyr Ala Gln Lys Met Glu Trp His Gly Val Gly Gly Gly Ser; Met Ala Gly Ser Leu Ser Thr Ile (SEQ ID NO: 82) Phe Asp Ala Gln Lys Ile Glu Trp His Val Gly Lys Gly Gly Ser; Met Ala Gln Gln Leu Pro Asp Ile (SEQ ID NO: 83) Phe Asp Ala Gln Lys Ile Glu Trp Arg Ile Ala Gly Gly Gly Ser; Met Ala Gln Arg Leu Phe His Ile (SEQ ID NO: 84) Leu Asp Ala Gln Lys Ile Glu Trp His Gly Pro Lys Gly Gly Ser; Met Ala Gly Cys Leu Gly Pro Ile (SEQ ID NO: 85) Phe Glu Ala Gln Lys Met Glu Trp Arg His Phe Val Gly Gly Ser; Met Ala Trp Ser Leu Lys Pro Ile (SEQ ID NO: 86) Phe Asp Ala Gln Lys Ile Glu Trp His Ser Pro Gly Gly Gly Ser; Met Ala Leu Gly Leu Thr Arg Ile (SEQ ID NO: 87) Leu Asp Ala Gln Lys Ile Glu Trp His Arg Asp Ser Gly Gly Ser; Met Ala Gly Ser Leu Arg Gln Ile (SEQ ID NO: 88) Leu Asp Ala Gln Lys Ile Glu Trp Arg Arg Pro Leu Gly Gly Ser, and; Met Ala Asp Arg Leu Ala Tyr Ile (SEQ ID NO: 89) Leu Glu Ala Gln Lys Met Glu Trp His Pro His Lys Gly Gly Ser.","These peptides, or fragments thereof which include SEQ ID NO 1 are suitable examples of peptides for use in producing fusion proteins in step (i).","In particular, the peptides used in the method of the invention to form the fusion protein have from 13 to 20 amino acids, and preferably about 15 amino acids.","A particularly preferred peptide for use in the fusion protein of the invention is a 15 amino acid peptide fragment of SEQ ID NO 78 shown above.","Specifically, a preferred peptide is of amino acid sequence SEQ ID NO 2: Gly Leu Asn Asp Ile Phe Glu Ala Gln (SEQ ID NO 2) Lys Ile Glu Trp His Glu.","This peptide is known as AviTag™ and DNA vectors encoding this sequence are available from Avidity Inc., sold under the trade names pAN4, pAN-5 and pAN-6 (which are suitable for producing fusion proteins in which the peptide of SEQ ID NO 2 is attached at the N terminus of the protein) and pAC-4, pAC-5 and pAC-6 (which are suitable for producing fusion proteins in which the peptide of SEQ ID NO 2 is attached at the C terminus of the protein).","The sequence of these vectors are shown hereinafter as SEQ ID NO 3, SEQ ID NO 4, SEQ ID NO 5, SEQ ID NO 6, SEQ ID NO 7 and SEQ ID NO 8 in FIGS.","7-12 respectively.","These vectors further include the ampicillin resistance gene bla to assist in cloning.","However the AviTag™ sequence can also be transferred into other vector systems.","Biotinylation can be effected in various ways, either in vivo or in vitro, for example by by co-expressing biotin ligase in the expression host, by adding biotin ligase to the cell lysate or by adding the biotin ligase to the purified protein.","In a particularly preferred embodiment, the method utilises the ability to enzymatically biotinylate a lysine residue in the fusion peptide in vivo prior to protein isolation from the cell lysate, by co-expressing biotin ligase in the expression host.","Usually when in vitro techniques are used, the expressed protein must first be isolated from the cell lysate and then chemically biotinylated in vitro by means well known in the art.","This results in loss of material and random biotinylation.","Proteins with multiple biotinylated sites have an unpredictable orientation and degree of binding onto the capture surface.","The advantage of the method of the invention is that all expressed proteins will be uniformly attached via the same residue on the same linker to the array Thus, suitably, the recombinant cell used in step (i) of the invention is engineered such that it also expresses a biotinylating enzyme and also contains biotin, such that step (ii) is effected in vivo in said cell as illustrated diagrammatically hereinafter in FIG.","1 .","DNA (1), which is suitably a cloned gene encoding an antigen or an antibody binding protein is sub-cloned into a vector (2) (such as pAN4, pAN-5, pAN-6 or pAC4, pAC-5 or pAC-6) which includes a sequence (3) encoding a peptide of SEQ ID NO.","1.The subcloned gene is then expressed in an expression system such as E. coli, which has been transformed with the vector as a fusion protein (4) comprising the antigen or antibody binding protein (5) fused to a peptide (6) of SEQ ID NO.","1.When expressed in-vivo in the presence of constitutively expressed biotin ligase, the lysine residue on the fusion peptide (6) is enzymatically biotinylated.","If the cell does not produce biotin, then it may be added to the culture medium in order to produce the desired result.","This reduces the number of steps involved in the process.","A particularly suitable host cell for use in the method of the invention are the AVB100, AVB 101 and AVB99 E. coli strains available from Avidity Inc., Denver, Colo., USA.","These strains all have the birA gene stably integrated into the chromosome so that they express biotin ligase.","In the case of AVB 100, overexpression of of BirA protein may be achieved by induction with L-arabinose.","The AVB101 E. coli B strain contains the pACYC184 ColE1 compatible plasmid that over-expresses biotin ligase, the elevated levels of Biotin Ligase in the cells result in complete biotinylation of fusion proteins in vivo.","An alternative host cell is strain AVB99 (Avidity Inc) which is an E. coli strain (XL1-Blue) containing a pACYC184 plasmid with an IPTG-inducible birA gene to overexpress biotin ligase (pBirAcm).","Fusion proteins produced in step (i) may also be isolated and biotinylated in vitro in the usual way.","The structure of the peptide of SEQ ID NO 1 is such that biotinylation will occur reliably at lysine adjacent X 6 within SEQ ID NO 1.In a preferred embodiment of the invention, the peptide comprising the amino acid of SEQ ID NO 1 is also used as a means of isolating the fusion protein in step (iii) of the method.","The technology for protein expression using recombinant DNA technology is well known in the art.","However, each protein that is expressed has a different amino acid sequence and many sequences are either difficult to express in the host of choice or their sequence is hydrophobic, and therefore insoluble, or is toxic to the host.","Even in the simplest bacterial expression systems, inclusion bodies are often formed that are difficult to disrupt while leaving the target protein in its native active state.","It is now common practise to fuse the target protein with another protein/peptide sequence (tag) to aid the purification process subsequent to expression.","Examples of such fusion expression systems are now widely used and have been commercialised by several suppliers.","Such fusion peptide sequences are attached to the amino or carboxyl terminal end of a protein sequence and are recognised by specific antibodies or affinity resins.","The expressed proteins must be solubilised from the cellular debris sometimes requiring harsh conditions including unphysiological pH values or use of chaotropic reagents and therefore the affinity purification process must be robust enough to function under such conditions.","By using this sequence as a means of isolating or purifying the expressed fusion protein, the need for additional purification tags is eliminated.","Thus this sequence has a dual purpose.","In some cases it may be desirable to use a further peptide sequence tag as a means of isolating or purifying the expressed fusion protein.","The sequence is preferably between 1 and 30 amino acids in length.","The peptide sequence tag sequence (20) may be located at the N-terminal or C-terminal region of the antigen or antibody binding protein as shown in FIG.","13 .","It is, however, preferably located at the opposite end of the antigen or antibody binding protein to which SEQ ID NO 1 is fused.","Where the additional peptide sequence tag is located on the same terminal region as SEQ ID NO 1, it is preferably fused to the free end of SEQ ID NO 1.Many peptide sequence tags are known in the art.","Examples of suitable peptide sequence tags for the purposes of the present invention are described in U.S. Pat.","No.","4,569,794A, and EP0 282 042B, the contents of which are herein incorporated by reference.","Preferably, the peptide sequence tag comprises at least one histidine amino acid.","Even more preferably the peptide sequence tag has the formula His-X in which X is selected from the group consisting of -Gly-, -His-, -Tyr-, -Gly-, -Trp-, -Val-, -Leu-, -Ser-, -Lys-, -Phe-, -Met-, -Ala-, -Glu-, -Ile-, -Thr-, -Asp-, -Asn-, -Gln-, -Arg-, -Cys- and -Pro-.","Alternatively the peptide sequence tag has the formula Y-His wherein Y is selected from -Gly-, -Ala-, -His- and -Tyr-.","Particularly suitable peptide sequence tags are described in EP 0 282 042B, and a preferred example is a hexa His tag.","In one embodiment of the method of the invention, step (iii) is effected using a further antibody or a binding fragment thereof, which is specific for the peptide of amino acid sequence including SEQ ID NO 1.The said further antibody may be raised using conventional techniques to the peptide (7) which includes an amino acid of SEQ ID NO 1.This method is illustrated diagrammatically in FIG.","2 .","The said further antibody is an anti-fusion antibody (8), which may be immobilised on a column, magnetic bead (9) or pipette tip, for example using a secondary antibody which is suitably an anti-species antibody (10) or other methods described in the literature, such as using an antibody binding protein such as Protein A, Protein G or Protein L, bound to the bead (9).","This approach is highly suited to automation and to the isolation of large numbers, but small quantities, of novel fusion proteins in parallel.","After separation from the cell lysate residue, the bound fusion protein (4) can subsequently be eluted by increasing the pH from 7.0 to 9.0.In an alternative embodiment, the fusion protein is isolated using a separation material which has some affinity for biotin but which releases the biotin fairly readily.","Suitably the separation material is a modified version of avidin or streptavidin, which has lower affinity for biotin than native avidin or streptavidin.","A particular example of such a material is a modified version of avidin marketed as CaptAvidin™ by by Molecular Probes (Eugene, Oreg., USA).","In this embodiment, the fusion protein is isolated from the cellular debris, detergents and salts etc from the culture medium, by lowering the pH of the cell lysis mixture to pH 6.0 followed by affinity purification with CaptAvidin™ attached to (a) magnetic beads or (b) pipette tips using conventional methods.","Bound fusion protein may then be eluted from the magnetic beads or mini columns by subsequently increasing the pH from 6.0 to 9.5.Prior to the step (iv), it is preferable to confirm the identity of the expressed fusion protein.","In one particular technique, a very low volume (10 μl) of the isolated fusion protein is removed from the microtitre plate.","The sample is digested by trypsin (using methods well known in the art).","The resultant peptide extract is desalted and concentrated using a ZipTip™ (Millipore, Mass., USA) or equivalent, before analysis via mass spectrometry.","Since the sequence of the fusion protein is known, identification by MALDI spectrometry to identify the peptides is usually sufficient to confirm the identity of the fusion protein.","This technique is widely used in protein research and is summarised by T. Rabilloud (Editor) Proteome Research: 2D gel electrophoresis and identification methods.","Furthermore, this technique can be automated and there are a number of commercially available systems from companies including Amersham Pharmacia Biotech, Bio Rad, AbiMed and Genomic Solutions (WO074852A1) that will perform this function.","Similarly, prior to step (iv), it is preferable that the concentration of each expressed protein should be normalised where possible to eliminate variation between elements.","Large variations in protein density cause difficulties in interpreting the data derived from such arrays (see Ekins, Clinical Chemistry (1998) 44:9 2015-2030, U.S. Pat.","No 5,807,755 and U.S. Pat.","No.","5,432,099 for a detailed discussion on the quantitative aspects of protein immunoassays and protein arrays and definitions of assay sensitivity).","Protein normalisation can be achieved by either determining the total protein concentration and or by including internal controls in the protocols.","In a particularly preferred embodiment of the method of the invention, the fusion tag is used as an internal control and is detected by an antibody with a high affinity for the peptide of amino acid sequence which includes SEQ ID NO 1 within the fusion protein.","Alternatively, the fusion protein can be expressed with a further peptide sequence tag and this can be used as an internal control.","Such a tag may be the said further peptide sequence tag such as a hexa His tag as discussed above, which is expressed as part of the biotinylated fusion protein.","The tagged version of this fusion protein may be detected through the use of an appropriate antibody such as an anti His tag antibody.","This can be done by performing a classic immunoassay sandwich simultaneously with, or during, a subsequent analysis of a biological sample using the array.","Using the antibody to the biotinylated fusion peptide, it is possible to determine the content of the fusion protein per μl and as a ratio of total protein present.","The methodology may be performed using a sheep polyclonal primary antibody and secondary antibody sandwich in which the secondary antibody is conjugated with fluorescent dye (e.g.","goat anti-mouse antibody conjugated to Alexa 488, Molecular Probes, Eugene, USA).","The fluorescent dye used is spectrally distinct from any used with the secondary antibody for the biological sample.","Both processes have been optimised for automation.","The avidin or steptavidin coated non-porous support used in step (iv) of the method of the invention is suitably a glass or plastics material.","Such supports are well suited to the production of small concentrated arrays.","This is important, since biological samples are generally very limited in volume, and thus very valuable.","A minimal surface area containing the targets is required for protein arrays, while still enabling the ability to achieve the required sensitivity of the assay is desirable.","In addition, high density of either antigen or antibody in the array produces better signal to noise ratios when used in an assay.","Furthermore, as compared to supports with porous surfaces including membranes, non-porous supports are more physically robust, are well suited to automation and have a lower background when imaged on fluorescent scanners.","These may be coated with avidin or streptavidin using conventional methods.","For example, the immobilisation of streptavidin to non-porous surfaces such as polystyrene multi-well plates is well known in the art.","In its most basic form, a solution of streptavidin is left in contact with the surface for some hours.","Un-bound protein is then removed by washing and the residual active moieties on the plastic surface blocked with BSA or an equivalent.","Although this approach may be passive, it is effective.","The non-covalent binding of streptavidin to polystyrene or nitrocellulose surfaces appears to be highly stable and resistant to elevated temperatures and high concentrations of chaotropic reagents, as described in WO98/37236.Avidin can be chemically attached to glass using the N-hydroxysuccinamide active ester of avidin as described by Manning, et al.","Biochemistry 16: 1364-1370 (1977) and can be attached to nylon via carbodiimide based coupling methods as described by Jasiewicz, et al.","Exp.","Cell Res.","100: 213-217 (1976).","In another method, high molecular weight compounds such as biotin-N-hydroxy-succinimide ester, N-biotinyl-6-aminocaproyl-N-hydroxysulfosuccinimide ester, sulfosuccinimidyl-2-(biotinamido)ethyl-1,3-dithiopropionate were biotinylated and used to coat a suitable surface.","Avidin or streptavidin was then coated in a second layer and was retained through binding the biotin linker attached to the high molecular weight compound as described in EP0620438.Attachment of streptavidin via a layer of biotin on the support surface was further developed in WO98/59243, which describes how biotin can be attached to a surface by chemical means or by light activation at 365 nm.","The benefit that this provides is that regions of the surface can be masked.","The elegance of these approaches is that biotin can be covalently bound to glass surfaces and will, in turn, non-covalently bind streptavidin only in those areas of the support that have been treated.","This enables patterns of streptavidin “acceptor” protein on the support to be manufactured if required.","In a preferred embodiment of the invention however, the entire surface of the non-porous support is coated with avidin or streptavidin, and then areas which are not required for binding are blocked, for example by addition of bovine serum albumin (BSA).","In this way, any non-specific interaction of fusion protein with the support is reduced.","Proteins that have been applied directly onto glass or plastic surfaces become non-covalently bound through interactions with charged groups on the solid surface the active moieties of the solid surface (typically silanol groups in glass or charged surface residues on polystyrene).","Such non-specific adsorption of antigens or antibodies onto the surfaces of glass and plastic significantly reduces their antigenicity and antigen binding capacity respectively.","The avidin or streptavidin layer therefore fulfils a dual role of firstly attaching the biotinylated fusion protein (non-biotinylated proteins that co-purify do not bind enabling a further purification step) and secondly, the dense layer of streptavidin shields the biotinylated fusion protein from undesirable non-specific interactions with the support surface.","When the fusion protein is applied to the avidin or streptavidin coating on the support in step (iv), very tight but non-covalent bonding occurs.","Preferably, the non-porous support is coated with streptavidin.","Biotin attachment to streptavidin is multivalent, providing a binding of very high capacity when compared to that of the antigen bound directly to the support surface.","Once the proteins have been applied to form the array, the bonding is strong enough to withstand extensive and stringent washing without appreciable loss of fusion protein.","This is illustrated in FIG.","3 .","Biotinylated fusion protein (4) is attached to the surface of the array support (12) via tight, non-covalent interaction with streptavidin (14).","In the preferred example, streptavidin (14) is covalently bound to the support material.","Sites on the array support material to which no streptavidin molecules are bound, are blocked by BSA or other surface modifiers (13).","Fusion proteins bind the steptavidin (14) via the biotin label (7) on the fusion peptide (6).","Furthermore, the avidin or streptavidin layer, whether attached directly to the surface of the support or via a biotinylated linker, is highly stable, is capable of being stored dry and can be heated and or treated with aggressive reagents without apparent loss of function (unlike most antigens and antibodies).","Further “acceptor” layers can be constructed on top of the foundation of the streptavidin layer if required.","These may comprise other antibody binding proteins known in the art.","The array will have the advantage of using the concentrating effect of the streptavidin, which has multivalent sites for biotin attachment.","This enables four times the biotin interaction with both antigen arrays and antibody arrays.","This allows higher densities of either antigen or antibody in each element of the array which in turn means that more elements can be assembled per mm 2 while achieving the same signal (see U.S. Pat.","No.","5,807,755 and U.S. Pat.","No.","5,432,099 for a discussion of the quantitative aspects) and less surface area of the solid support is utilised.","The advantage is that less biological sample is thus required.","Where the array is to consist of antigens arrayed on a microscope format, it suitably contains a large number of these, for example from 3-10,000 different fusion proteins.","These may be generated or obtained from various sources, depending upon the intended nature and target for the analysis to be conducted using the array.","Preferably, however, each protein will be expressed in the form of two fusion proteins, one with the peptide including SEQ ID NO 1 attached to the C-terminal, and one with the peptide including SEQ ID NO 1 attached to the N-terminal.","In this way, the relative antigenicities of each version of the fusion protein to a complementary antibody can be assessed.","The uses of antibody binding proteins such as Proteins A, G and L have been extensively reported.","The binding of such proteins to antibodies is sufficiently tight to enable use in separation and detection techniques.","These proteins are known to bind to the conserved regions of various classes of antibody.","When the method of the invention is used to produce antibody arrays, antibody attachment is achieved by capture of the antibody via use of a layer of antibody binding proteins fused to a peptide which comprises SEQ ID NO 1.This layer preferably comprises of a mixture of biotinylated tagged Proteins A, G and L, and more preferably, some of which are fused at the C-terminal end, and some of which are labelled at the N-terminal end.","By creating a mixture of antibody-binding proteins, a universal acceptor is created enabling the attachment of virtually any antibody, polyclonal, monoclonal, full-chain fragments, single chain antibodies and phage with antibody activity into which a Protein A, G or L site is present or has been engineered.","Any antibody can be incorporated into the array without the need to pre-process or modify the antibody.","When such antibody binding proteins are applied to an avidin or streptavidin coated surface in step (iv) a very high density of bound protein (biotinylated tagged protein binding to multivalent streptavidin) results.","This method has the advantage that only one amino acid residue is biotinylated, and this is part of the fusion peptide, leaving the antibody binding sites on the lectins available.","These antibody binding proteins effectively create a second layer on the solid surface.","This layer comprising of bound fusion tagged Proteins A, G and L, acts as a universal acceptor surface for any antibody ( FIG.","6 ) without the need for direct biotinylation of the antibody.","This saves time, antibody and eliminates the possible degradation of the antibody's binding to its corresponding antigen.","In a preferred embodiment, a molar excess of antibodies is pre-mixed with biotinylated peptide antibody-binding protein (e.g.","Proteins A, G or L) fusion and incubated for up to 15 minutes.","This antibody-antibody binding protein mixture is then applied directly to the streptavidin covered array support.","Alternatively, the biotinylated antibody binding protein fusion may first be applied to the streptavidin covered array support.","Individual antibodies are then applied to the surface of the coated support to form an array.","The array produced by either method comprises very discrete spots with minimal observable diffusion, leading to a good array for assay purposes.","The array obtained using the method of the invention is suitably used in methods for detecting binding between antigens and antibodies.","Thus in a second aspect, the invention provides a method of detecting binding between an antibody and an antigen, said method comprising the steps of (vi) applying to the array obtained using a method of the first aspect a sample which contains or is suspected of containing an antibody in the case of an array of step (v)(a), or an antigen in the case of the array of step (v)(b); and (vii) detecting bound antibody or antigen on the support.","Steps (vi) and (vii) of the method of the invention are suitably carried out in a conventional manner, using well known immunlogical techniques such as ELISAs, including sandwich ELISAs using labelled and in particular fluorescently labelled antibodies.","This is illustrated in the case of an antigen array in FIG.","4 .","Antigens (4) bound to the array substrate (12) via streptavidin (14) are detected with a suitable primary antibody (15).","The signal is amplified using a suitable secondary antibody (16) conjugated to a label (17).","The label in the preferred embodiment is a fluorescent dye, such as Alexa 488, but may be any number of other types of label that are known in the art.","Suitably the protein array continues to be monitored for quality and in particular the density of the protein during use of protein analysis devices.","This is achieved in accordance with a preferred embodiment of the invention by using the peptide which comprises SEQ ID NO 1 or the further peptide sequence tag such as the hexa His Tag mentioned above, or any other suitable tag which performs this function as an internal standard, in a manner similar to that described above for pre-array protein normalisation.","Once the antigens from numerous protein preparations have been arrayed onto the support surface, the array can then be used to assess antibody quality (see WO 99/39210) or can be used to determine antibody titre in serum samples (Joos et al Electrophoresis 2000, 21, 2641-2650).","In these instances, the relative amounts of protein between the different elements in an array can be determined by adding an internal standard to the primary sample.","The internal standard that is preferred is a sheep polyclonal antibody raised against the fusion peptide.","This is spiked into the primary antibody solution (either antibody or serum) and is detected by an anti-sheep secondary antibody conjugated with a suitable fluorescent dye that is spectrally distinct from the labelled secondary antibody to the primary sample.","A two-colour image is generated using a commercially available slide imager and the signal for each element is normalised to the signal resulting from the fusion protein.","Combined with pre-array protein content normalisation, arrays of considerable consistency can be generated.","Preferably at least some and most preferably all of the steps of the process described above are operated automatically to increase throughput and reduce labour time and costs.","Creating antigen arrays with many novel proteins means proteins must be attached with the minimum number of steps if the process is to be viable.","The use of a peptide which is readily and specifically biotinylated and which can act not only as a binding protein for assay purposes, but also as a purification means and an internal control for monitoring quality, provides just such a method.","The method of the invention allows diverse collections of proteins to be attached with universal procedures, a minimum number of steps and maximum predictability of orientation.","The method is suitable for operation on a large-scale, for example in high-throughput screening.","In a third aspect the invention provides a protein array on a non-porous support, obtained using the method of the first aspect of the invention.","Some elements used in the above-described methods are novel and therefore form further aspects of the invention.","In particular, in a fourth aspect, the invention provides a fusion protein comprising an antibody binding protein fused at the N— or C-terminus to a peptide of 13 to 50 amino acids which comprises SEQ ID NO 1, such as a peptide of SEQ ID NO 2.In particular, the antibody binding protein is Protein A, G or L and preferably a mixture thereof.","The fusion protein may additionally comprise a further peptide sequence tag such as the hexa His tag or another suitable sequence tag, which are known to those skilled in the art as discussed above.","Such sequence tag may be located at the N or the C-terminus of the antigen or antibody binding protein.","It is, however, preferably located at the opposite end of the antigen or antibody binding protein to which the amino acid sequence of SEQ ID NO 1 is fused.","Where it is located at the same terminal region as SEQ ID NO 1, the sequence tag is fused to the free end of SEQ ID NO 1.A fifth aspect of the invention comprises a nucleic acid sequence which encodes the fusion protein of the fourth aspects.","In particular in this case, the sequence which encodes the peptide is suitably of SEQ ID NO 9.","(SEQ ID NO 9) GGCCTGAACGACATCTTCGAGGCTCAGAAAATCGAATGGCACGAA"],["FIELD OF THE INVENTION The present invention relates to a method of producing arrays for conducting protein analysis, in particular of antibodies, antigens or antibody binding proteins, to protein arrays produced, methods of conducting analysis using them and novel entities incorporated in them.","More specifically, the process relates to a method of producing a range of antibodies and/or antigens and immobilising these in an array, for use in protein or binding analysis.","BACKGROUND The concept of attaching a number of different proteins to surface supports to form an “array” of proteins has been widely described in the literature (see for example EP0063810, WO84/03151, U.S. Pat.","No.","5,143,854).","Recently, there has been a growing interest in the concept of manufacturing devices whereby large numbers of proteins of various classes are arrayed onto different types of solid supports.","Examples include antigen, antibody, protein (protein-protein interaction) and functional enzymes arrays.","The background to the technology, and the potential uses for such devices, are thoroughly catalogued in the literature (Joos et al Electrophoresis 2000, 21, 2641-2650, Haab et al Genome Biology 2000 1(6), Borrebaeck Immunology Today, August 2000) and examples of potential utility can be found in a number of recent patent applications including WO 00/07024, WO 99/40434, WO 99/39210 and WO00/54046.The concept of creating antigen arrays was described in EP 0063810 in 1982.It was reported that antigens and antibodies could be bound to a porous solid support enabling an unlimited number of antibody-antigen interactions to be conducted simultaneously.","To make antigen arrays, antigens were simply aliquoted in very small volumes onto nitrocellulose membranes or similar supports, allowed to adsorb and then probed with the corresponding antibodies.","As with Enzyme Linked Immunosorbent Assay (ELISA) protocols performed in solution or in plastic plates, non-specific interactions were blocked with Bovine Serum Albumin (BSA), and this is now standard practise.","It was also reported that the elements (or spots) of the array did not diffuse and were adsorbed tightly onto the membrane.","It should be noted, however, that the dimensions for these elements were considerably larger than those obtained in micro array device systems.","EP 0063810 describes how the protein arrays could be made by aliquoting proteins by hand, using mechanical procedures including a “charged drop” or lithographic process.","In this manner elements with a diameter of less than 500 microns (compared with 100 microns that can be achieved with current automated array systems) were produced.","However, one of the main disadvantages associated with the use of membranes as opposed to non-porous surfaces is that the elements tend to diffuse through the support material unless there is immediate binding.","Attempts were made to overcome this problem.","U.S. Pat.","No.","4,496,654 describes use of porous surfaces such as paper disks which were treated with streptavidin (which is adsorbed onto the surface) enabling arrays of biotinylated antibodies to be arranged in any desired pattern.","Following blocking with BSA, the paper discs could be probed with the antigen (exemplified with human chorionic gonadotropin) which could then be detected with an enzyme assay.","The biotinylated antibody immediately bound very tightly to the surface of the paper reducing diffusion of the spots.","To achieve this using an “acceptor” surface such as an avidin or streptavidin coated surface, requires that each antibody and antigen, which is attached to the array, must be biotinylated prior to attachment to the array with no guarantee that this process will not impair its avidity (or antigenicity if an antigen is used) compared with the native protein.","Non-porous surfaces also have the disadvantage that they are not as robust as solid surfaces, including various types of glass or plastics, and so cannot be washed or treated as stringently.","For antigen and antibody arrays, it has been found however that attaching a protein to a solid surface generally leads to a reduction in antigenicity of the antigen and avidity of the antibody compared with that observed when the antigen or antibody is in free solution.","Previous attempts (see WO84/03 151 and Haab et al 2000 supra.)","to immobilise antigens and antibodies were not greatly successful.","WO84/03 151 describes that antibodies can be applied directly onto glass surfaces such as a microscope cover slip and dried.","When blocked and then exposed to antigens, in this case in the form of whole cells, the antigens were captured by the array.","However, WO84/03 151 further describes that these antigens needed to be added at a higher concentration compared with the equivalent ELISA performed in solution.","It was also noted that the antibodies had to be “highly enriched in order to achieve a sufficiently dense antibody coat for the desired cell adherence”.","It also took considerable time for the antibodies to be adsorbed onto the glass surface.","Other approaches for the direct immobilisation of antigens and antibodies have been reported.","One approach was to first adsorb calcium phosphate in the form of hydroxyapatite onto filter papers onto which proteins were bound by ionic interaction as described in U.S. Pat.","No.","5,827,669.These inventors reported that this was not effective for acidic proteins and that the antibodies suffered from “bad orientation” onto the Ca/phosphate layer.","Success was, however, reported when this method was used in immobilising streptavidin.","Another method for immobilising proteins to solid, non-porous surfaces included attaching them using an adhesive polyphenolic protein isolated from muscles as described in U.S. Pat.","No.","5,817,470.By coating solid surfaces, such as a polystyrene multi-well plate with polyphenolic protein, various antigens could be bound to the treated support and detected in an ELISA sandwich comprising of a primary antibody followed by a secondary antibody conjugated to an enzyme.","However, the inventors conceded that the procedure was limited by the amount of antigen bound or adsorbed to the solid surface.","The final amount of antigen strongly bound to the surface of the plate varied depending on a number of factors such as the molecular characteristics of the antigens, the properties of the solid support, the concentration of the antigen in the solution as well as the characteristics of the buffer used to dissolve the antigen used to coat or to activate the surface.","In general, only a small fraction of the antigen present in the coating solution was adsorbed to the surface.","Direct attachment of antibodies and antigens to non-porous surfaces was also been attempted with a collection of 113 antibodies and their corresponding antigens (Haab et al, 2000 Genome Biology 1(6)).","By exploiting technology developed for DNA microarrays, glass slides coated with poly-s-lysine were used to immobilise both antigens in one experiment and antibodies in another.","The results reported showed that only 50% of the arrayed antigens and 20% of the arrayed antibodies, provided specific and accurate measurements of their cognate ligands at or below concentrations of 1.60 μl/ml and 0.34 μg/ml respectively.","The high failure rate in binding antibodies to solid surfaces would not be acceptable for a large-scale antibody array manufacturing programme.","This supports the view that direct attachment of antigens and antibodies is an unsuitable technique to retain antibody/antigen functionality if protein arrays are to fulfil their potential.","The use of coatings such as avidin and streptavidin as binders for biotin labelled proteins is well known for use in conjunction with many proteins.","The proteins are generally isolated first, and then biotinylated.","Biotin can be conjugated to the protein at any or all active lysine sites contained within it.","Thus, when antigens or antibodies are biotinylated in this way, biotin groups may be present at their N-terminal groups and at any number of potential active lysine residues over their surface.","This means that they will adopt any number of different orientations once bound to the streptavidin layer and so the binding properties will be diverse.","Furthermore, access to the antigen or antibody immobilised via streptavidin will be reduced by steric hindrance, leading to generally inadequate assay.","It has been found that it is possible to reduce the steric hindrance and increase the sensitivity of the immunoassay by including a linker between the antigen/antibody and the biotinylated site.","U.S. Pat.","No.","5,811,246 describes how small synthetic peptides used in either immunoassays or for raising antisera can be linked to a “carrier” protein such as avidin or streptavidin via a linker such as various bradykinin derivatives.","This has several advantages.","Firstly, the condensation reaction between the free N-terminal group on the peptide and the linker preserves the charged residues essential for recognition by an antibody (immunoassay) or to elicit an immune response (immunisation).","Secondly, the bradykinin linker can then be biotinylated in such a way as to preserve the free charged groups on the small peptide.","In each case, the presence of the linker appears to promote a more sensitive immunoassay and an improved immune response when used as an immunising agent.","This use of a bradykinin derivative in this way however introduces further steps and complications into the process.","Biotinylated peptides fused to peptides or proteins of interest are described in U.S. Pat.","Nos.","5,723,584, 5,874,239 and 5,932,433, and further in Beckett et a!.","Protein Sci.","(1999) 8(4) 921-9.These peptides are used in order to biotinylate recombinant proteins, so as to allow rapid purification, immobilization, labelling and detection thereof.","It is not suggested that these peptides should be used in particular with antigens or antibody binding proteins, or that they should be formulated in arrays.","The present applicants have found that the peptides used in these patents allow the production of very good antigen or antibody arrays, which can be efficiently produced on non-porous supports whilst substantially retaining the binding avidity of these proteins.","SUMMARY OF THE INVENTION According to a first aspect of the present invention there is provided a method of forming an array of proteins selected from antigens or antibodies; said method comprising the steps of (i) expressing in a recombinant cell, a fusion protein which comprises either (a) an antigen or (b) an antibody binding protein, fused to a peptide having up to 50 amino acids, which peptide comprised amino acid sequence of SEQ ID NO 1 LX1X2IX3X4X5X6KX7X8X9X10 (SEQ ID NO 1) where X1 is a naturally occurring amino acid, X2 is any naturally occurring amino acid other than leucine, valine, isoleucine, tryptophan, phenylalanine or tyrosine, X3 is phenylalanine or leucine, X4 is glutamine or asparagine, X5 is alanine, glycine, serine or threonine, X6 is glycine or methionine, X7 is isoleucine, methionine or valine, X8 is glutamine, leucine, valine, tyrosine or isoleucine, X9 is tryptophan, tyrosine, valine, phenylalanine, leucine or isoleucine and X10 is any naturally occurring amino acid other than asparagine or glutamine; where said peptide is capable of being biotinylated by a biotin ligase at the lysine residue adjacent to X6; (ii) biotinylating said peptide of the fusion protein at the lysine residue adjacent X6; (iii) isolating the biotinylated fusion protein; (iv) applying the biotinylated fusion protein to an avidin or streptavidin coated non-porous support; (v) forming an array of at least three different proteins on the support by either (a) where the fusion protein comprises an antigen, carrying out steps (i) to (iv) the desired number of times to form an antigen array; or (b) where the fusion protein comprises an antibody binding protein, applying to said protein, either prior to or after step (iv) a plurality of different antibodies or binding fragments thereof.","The applicants have found that by using a fusion of the antigen or antibody binding protein to a peptide of SEQ ID NO 1, these proteins may be immobilised onto solid surfaces, whilst substantially maintaining the antigenicity of proteins, or the binding capabilities of the antibody binding proteins.","This may be because the fusion peptide is biotinylated rather than the protein itself, and so there is less disruption of the protein's antigenicity when attached to the support surface.","In addition, the peptide including SEQ ID NO 1 appears to reduce steric hindrance to enable interaction between antigen and antibody.","By ensuring that the peptide linker is attached at a terminal region of the protein, and contains the biotinylation site, sites on the protein which are essential for function appear to be largely unaffected.","This combination is particularly advantageous in the context of methods of analysis using antigens or antibody arrays.","The method described herein represents the first time that the mode of attachment of proteins to non-porous surfaces (step vi), the mode of protein isolation from cell lysate (step iii) and the method of biotinylation (step ii) utilise the same fusion peptide.","Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.","As used herein the expression “antibody binding protein” refers to proteins which are known to bind to regions of antibodies, or to mixtures of these.","Examples of such proteins include Protein A, Protein L and Protein G Antibody binding proteins are used in accordance with the invention in the production of antibody arrays.","The antibodies are bound by antibody binding proteins, such as Proteins A, G and/or L or a mixture of one or more of these, which are themselves anchored via the linker to the streptavidin coating on the support surface.","While biotinylated versions of native Protein A, G and L are commercially available and can be attached to the streptavidin coating on the support surface, the applicants have found that by fusing these proteins to biotinylated tags in accordance with the present invention at the C and/or N-terminals, highly effective binding of antibodies of various types was achieved.","This may also be the result of reduced steric factors, or that the binding sites on the proteins are all readily available.","In addition, by using the method of the invention, the biotinylated fusion protein is immediately captured on application to the avidin or streptavidin coated support in step (iv) leading to very discrete spots of protein on the support, with minimal observable diffusion.","Particular examples of peptides having up to 50 amino acids, which peptide comprises an amino acid sequence of SEQ ID NO 1 are listed in U.S. Pat.","No.","5,723,584, U.S. Pat.","No.","5,874,239 and U.S. Pat.","No.","5,932,433, the content of which are incorporated herein by reference.","Examples of peptides provided in these references are listed below: Leu Glu Glu Val Asp Ser Thr Ser (SEQ ID NO: 14) Ser Ala Ile Phe Asp Ala Met Lys Met Val Trp Ile Ser Pro Thr Glu Phe Arg; Gln Gly Asp Arg Asp Glu Thr Leu (SEQ ID NO: 15) Pro Met Ile Leu Arg Ala Met Lys Met Glu Val Tyr Asn Pro Gly Gly His Glu Lys; Ser Lys Cys Ser Tyr Ser His Asp (SEQ ID NO: 16) Leu Lys Ile Phe Glu Ala Gln Lys Met Leu Val His Ser Tyr Leu Arg Val Met Tyr Asn Tyr; Met Ala Ser Ser Asp Asp Gly Leu (SEQ ID NO: 17) Leu Thr Ile Phe Asp Ala Thr Lys Met Met Phe Ile Arg Thr; Ser Tyr Met Asp Arg Thr Asp Val (SEQ ID NO: 18) Pro Thr Ile Leu Glu Ala Met Lys Met Glu Leu His Thr Thr Pro Trp Ala Cys Arg; Ser Phe Pro Pro Ser Leu Pro Asp (SEQ ID NO: 19) Lys Asn Ile Phe Glu Ala Met Lys Met Tyr Val Ile Thr; Ser Val Val Pro Glu Pro Gly Trp (SEQ ID NO: 20) Asp Gly Pro Phe Glu Ser Met Lys Met Val Tyr His Ser Gly Ala Gln Ser Gly Gln; Val Arg His Leu Pro Pro Pro Leu (SEQ ID NO: 21) Pro Ala Leu Phe Asp Ala Met Lys Met Glu Phe Val Thr Ser Val Gln Phe; Asp Met Thr Met Pro Thr Gly Met (SEQ ID NO: 22) Thr Lys Ile Phe Glu Ala Met Lys Met Glu Val Ser Thr; Ala Thr Ala Gly Pro Leu His Glu (SEQ ID NO: 23) Pro Asp Ile Phe Leu Ala Met Lys Met Glu Val Val Asp Val Thr Asn Lys Ala Gly Gln; Ser Met Trp Glu Thr Leu Asn Ala (SEQ ID NO: 24) Gln Lys Thr Val Leu Leu; Ser His Pro Ser Gln Leu Met Thr (SEQ ID NO: 25) Asn Asp Ile Phe Glu Gly Met Lys Met Leu Tyr His; Thr Ser Glu Leu Ser Lys Leu Asp (SEQ ID NO: 27) Ala Thr Ile Phe Ala Ala Met Lys Met Gln Trp Trp Asn Pro Gly; Val Met Glu Thr Gly Leu Asp Leu (SEQ ID NO: 28) Arg Pro Ile Leu Thr Gly Met Lys Met Asp Trp Ile Pro Lys; Leu His His Ile Leu Asp Ala Gln (SEQ ID NO: 30) Lys Met Val Trp Asn His Arg; Pro Gln Gly Ile Phe Glu Ala Gln (SEQ ID NO: 31) Lys Met Leu Trp Arg Ser; Leu Ala Gly Thr Phe Glu Ala Leu (SEQ ID NO: 32) Lys Met Ala Trp His Glu His; Leu Asn Ala Ile Phe Glu Ala Met (SEQ ID NO: 33) Lys Met Glu Tyr Ser Gly; Leu Gly Gly Ile Phe Glu Ala Met (SEQ ID NO: 34) Lys Met Glu Leu Arg Asp; Leu Leu Arg Thr Phe Glu Ala Met (SEQ ID NO: 35) Lys Met Asp Trp Arg Asn Gly; Leu Ser Thr Ile Met Glu Gly Met (SEQ ID NO: 36) Lys Met Tyr Ile Gln Arg Ser; Leu Ser Asp Ile Phe Glu Ala Met (SEQ ID NO: 37) Lys Met Val Tyr Arg Pro Cys; Leu Glu Ser Met Leu Glu Ala Met (SEQ ID NO: 38) Lys Met Gln Trp Asn Pro Gln; Leu Ser Asp Ile Phe Asp Ala Met (SEQ ID NO: 39) Lys Met Val Tyr Arg Pro Gln; Leu Ala Pro Phe Phe Glu Ser Met (SEQ ID NO: 40) Lys Met Val Trp Arg Glu His; Leu Lys Gly Ile Phe Glu Ala Met (SEQ ID NO: 41) Lys Met Glu Tyr Thr Ala Met; Leu Glu Gly Ile Phe Glu Ala Met (SEQ ID NO: 42) Lys Met Glu Tyr Ser Asn Ser; Leu Leu Gln Thr Phe Asp Ala Met (SEQ ID NO: 43) Lys Met Glu Trp Leu Pro Lys; Val Phe Asp Ile Leu Glu Ala Gln (SEQ ID NO: 44) Lys Val Val Thr Leu Arg Phe; Leu Val Ser Met Phe Asp Gly Met (SEQ ID NO: 45) Lys Met Glu Trp Lys Thr Leu; Leu Glu Pro Ile Phe Glu Ala Met (SEQ ID NO: 46) Lys Met Asp Trp Arg Leu Glu; Leu Lys Glu Ile Phe Glu Gly Met (SEQ ID NO: 47) Lys Met Glu Phe Val Lys Pro; Leu Gly Gly Ile Glu Ala Gln Lys (SEQ ID NO: 48) Met Leu Leu Tyr Arg Gly Asn; Arg Pro Val Leu Glu Asn Ile Phe (SEQ ID NO: 50) Glu Ala Met Lys Met Glu Val Trp Lys Pro; Arg Ser Pro Ile Ala Glu Ile Phe (SEQ ID NO: 51) Glu Ala Met Lys Met Glu Tyr Arg Glu Thr; Gln Asp Ser Ile Met Pro Ile Phe (SEQ ID NO: 52) Glu Ala Met Lys Met Ser Trp His Val Asn; Asp Gly Val Leu Phe Pro Ile Phe (SEQ ID NO: 53) Glu Ala Met Lys Met Ile Arg Leu Glu Thr; Val Ser Arg Thr Met Thr Asn Phe (SEQ ID NO: 54) Glu Ala Met Lys Met Ile Tyr His Asp Leu; Asp Val Leu Leu Pro Thr Val Phe (SEQ ID NO: 55) Glu Ala Met Lys Met Tyr Ile Thr Lys; Pro Asn Asp Leu Glu Arg Ile Phe (SEQ ID NO: 56) Asp Ala Met Lys Ile Val Thr Val His Ser; Thr Arg Ala Leu Leu Glu Ile Phe (SEQ ID NO: 57) Asp Ala Gln Lys Met Leu Tyr Gln His Leu; Arg Asp Val His Val Gly Ile Phe (SEQ ID NO: 58) Glu Ala Met Lys Met Tyr Thr Val Glu Thr; Gly Asp Lys Leu Thr Glu Ile Phe (SEQ ID NO: 59) Glu Ala Met Lys Ile Gln Trp Thr Ser Gly; Leu Glu Gly Leu Arg Ala Val Phe (SEQ ID NO: 60) Glu Ser Met Lys Met Glu Leu Ala Asp Glu; Val Ala Asp Ser His Asp Thr Phe (SEQ ID NO: 61) Ala Ala Met Lys Met Val Trp Leu Asp Thr; Gly Leu Pro Leu Gln Asp Ile Leu (SEQ ID NO: 62) Glu Ser Met Lys Ile Val Met Thr Ser Gly; Arg Val Pro Leu Glu Ala Ile Phe (SEQ ID NO: 63) Glu Gly Ala Lys Met Ile Trp Val Pro Asn Asn; Pro Met Ile Ser His Lys Asn Phe (SEQ ID NO: 64) Glu Ala Met Lys Met Lys Phe Val Pro Glu; Lys Leu Gly Leu Pro Ala Met Phe (SEQ ID NO: 65) Glu Ala Met Lys Met Glu Trp His Pro Ser; Gln Pro Ser Leu Leu Ser Ile Phe (SEQ ID NO: 66) Glu Ala Met Lys Met Gln Ala Ser Leu Met; Leu Leu Glu Leu Arg Ser Asn Phe (SEQ ID NO: 67) Glu Ala Met Lys Met Glu Trp Gln Ile Ser; Asp Glu Glu Leu Asn Gln Ile Phe (SEQ ID NO: 68) Glu Ala Met Lys Met Tyr Pro Leu Val His Val Thr Lys; Ser Asn Leu Val Ser Leu Leu His (SEQ ID NO: 70) Ser Gln Lys Ile Leu Trp Thr Asp Pro Gln Ser Phe Gly; Leu Phe Leu His Asp Phe Leu Asn (SEQ ID NO: 71) Ala Gln Lys Val Glu Leu Tyr Pro Val Thr Ser Ser Gly; Ser Asp Ile Asn Ala Leu Leu Ser (SEQ ID NO: 72) Thr Gln Lys Ile Tyr Trp Ala His; Met Ala Ser Ser Leu Arg Gln Ile (SEQ ID NO: 73) Leu Asp Ser Gln Lys Met Glu Trp Arg Ser Asn Ala Gly Gly Ser; Met Ala His Ser Leu Val Pro Ile (SEQ ID NO: 75) Phe Asp Ala Gln Lys Ile Glu Trp Arg Asp Pro Phe Gly Gly Ser; Met Gly Pro Asp Leu Val Asn Ile (SEQ ID NO: 76) Phe Glu Ala Gln Lys Ile Glu Trp His Pro Leu Thr Gly Gly Ser; Met Ala Phe Ser Leu Arg Ser Ile (SEQ ID NO: 77) Leu Glu Ala Gln Lys Met Glu Leu Arg Asn Thr Pro Gly Gly Ser; Met Ala Gly Gly Leu Asn Asp Ile (SEQ ID NO: 78) Phe Glu Ala Gln Lys Ile Glu Trp His Glu Asp Thr Gly Gly Ser; Met Ser Ser Tyr Leu Ala Pro Ile (SEQ ID NO: 79) Phe Glu Ala Gln Lys Ile Glu Trp His Ser Ala Tyr Gly Gly Ser; Met Ala Lys Ala Leu Gln Lys Ile (SEQ ID NO: 80) Leu Glu Ala Gln Lys Met Glu Trp Arg Ser His Pro Gly Gly Ser; Met Ala Phe Gln Leu Cys Lys Ile (SEQ ID NO: 81) Phe Tyr Ala Gln Lys Met Glu Trp His Gly Val Gly Gly Gly Ser; Met Ala Gly Ser Leu Ser Thr Ile (SEQ ID NO: 82) Phe Asp Ala Gln Lys Ile Glu Trp His Val Gly Lys Gly Gly Ser; Met Ala Gln Gln Leu Pro Asp Ile (SEQ ID NO: 83) Phe Asp Ala Gln Lys Ile Glu Trp Arg Ile Ala Gly Gly Gly Ser; Met Ala Gln Arg Leu Phe His Ile (SEQ ID NO: 84) Leu Asp Ala Gln Lys Ile Glu Trp His Gly Pro Lys Gly Gly Ser; Met Ala Gly Cys Leu Gly Pro Ile (SEQ ID NO: 85) Phe Glu Ala Gln Lys Met Glu Trp Arg His Phe Val Gly Gly Ser; Met Ala Trp Ser Leu Lys Pro Ile (SEQ ID NO: 86) Phe Asp Ala Gln Lys Ile Glu Trp His Ser Pro Gly Gly Gly Ser; Met Ala Leu Gly Leu Thr Arg Ile (SEQ ID NO: 87) Leu Asp Ala Gln Lys Ile Glu Trp His Arg Asp Ser Gly Gly Ser; Met Ala Gly Ser Leu Arg Gln Ile (SEQ ID NO: 88) Leu Asp Ala Gln Lys Ile Glu Trp Arg Arg Pro Leu Gly Gly Ser, and; Met Ala Asp Arg Leu Ala Tyr Ile (SEQ ID NO: 89) Leu Glu Ala Gln Lys Met Glu Trp His Pro His Lys Gly Gly Ser.","These peptides, or fragments thereof which include SEQ ID NO 1 are suitable examples of peptides for use in producing fusion proteins in step (i).","In particular, the peptides used in the method of the invention to form the fusion protein have from 13 to 20 amino acids, and preferably about 15 amino acids.","A particularly preferred peptide for use in the fusion protein of the invention is a 15 amino acid peptide fragment of SEQ ID NO 78 shown above.","Specifically, a preferred peptide is of amino acid sequence SEQ ID NO 2: Gly Leu Asn Asp Ile Phe Glu Ala Gln (SEQ ID NO 2) Lys Ile Glu Trp His Glu.","This peptide is known as AviTag™ and DNA vectors encoding this sequence are available from Avidity Inc., sold under the trade names pAN4, pAN-5 and pAN-6 (which are suitable for producing fusion proteins in which the peptide of SEQ ID NO 2 is attached at the N terminus of the protein) and pAC-4, pAC-5 and pAC-6 (which are suitable for producing fusion proteins in which the peptide of SEQ ID NO 2 is attached at the C terminus of the protein).","The sequence of these vectors are shown hereinafter as SEQ ID NO 3, SEQ ID NO 4, SEQ ID NO 5, SEQ ID NO 6, SEQ ID NO 7 and SEQ ID NO 8 in FIGS.","7-12 respectively.","These vectors further include the ampicillin resistance gene bla to assist in cloning.","However the AviTag™ sequence can also be transferred into other vector systems.","Biotinylation can be effected in various ways, either in vivo or in vitro, for example by by co-expressing biotin ligase in the expression host, by adding biotin ligase to the cell lysate or by adding the biotin ligase to the purified protein.","In a particularly preferred embodiment, the method utilises the ability to enzymatically biotinylate a lysine residue in the fusion peptide in vivo prior to protein isolation from the cell lysate, by co-expressing biotin ligase in the expression host.","Usually when in vitro techniques are used, the expressed protein must first be isolated from the cell lysate and then chemically biotinylated in vitro by means well known in the art.","This results in loss of material and random biotinylation.","Proteins with multiple biotinylated sites have an unpredictable orientation and degree of binding onto the capture surface.","The advantage of the method of the invention is that all expressed proteins will be uniformly attached via the same residue on the same linker to the array Thus, suitably, the recombinant cell used in step (i) of the invention is engineered such that it also expresses a biotinylating enzyme and also contains biotin, such that step (ii) is effected in vivo in said cell as illustrated diagrammatically hereinafter in FIG.","1.DNA (1), which is suitably a cloned gene encoding an antigen or an antibody binding protein is sub-cloned into a vector (2) (such as pAN4, pAN-5, pAN-6 or pAC4, pAC-5 or pAC-6) which includes a sequence (3) encoding a peptide of SEQ ID NO.","1.The subcloned gene is then expressed in an expression system such as E. coli, which has been transformed with the vector as a fusion protein (4) comprising the antigen or antibody binding protein (5) fused to a peptide (6) of SEQ ID NO.","1.When expressed in-vivo in the presence of constitutively expressed biotin ligase, the lysine residue on the fusion peptide (6) is enzymatically biotinylated.","If the cell does not produce biotin, then it may be added to the culture medium in order to produce the desired result.","This reduces the number of steps involved in the process.","A particularly suitable host cell for use in the method of the invention are the AVB100, AVB 101 and AVB99 E. coli strains available from Avidity Inc., Denver, Colo., USA.","These strains all have the birA gene stably integrated into the chromosome so that they express biotin ligase.","In the case of AVB 100, overexpression of of BirA protein may be achieved by induction with L-arabinose.","The AVB101 E. coli B strain contains the pACYC184 ColE1 compatible plasmid that over-expresses biotin ligase, the elevated levels of Biotin Ligase in the cells result in complete biotinylation of fusion proteins in vivo.","An alternative host cell is strain AVB99 (Avidity Inc) which is an E. coli strain (XL1-Blue) containing a pACYC184 plasmid with an IPTG-inducible birA gene to overexpress biotin ligase (pBirAcm).","Fusion proteins produced in step (i) may also be isolated and biotinylated in vitro in the usual way.","The structure of the peptide of SEQ ID NO 1 is such that biotinylation will occur reliably at lysine adjacent X6 within SEQ ID NO 1.In a preferred embodiment of the invention, the peptide comprising the amino acid of SEQ ID NO 1 is also used as a means of isolating the fusion protein in step (iii) of the method.","The technology for protein expression using recombinant DNA technology is well known in the art.","However, each protein that is expressed has a different amino acid sequence and many sequences are either difficult to express in the host of choice or their sequence is hydrophobic, and therefore insoluble, or is toxic to the host.","Even in the simplest bacterial expression systems, inclusion bodies are often formed that are difficult to disrupt while leaving the target protein in its native active state.","It is now common practise to fuse the target protein with another protein/peptide sequence (tag) to aid the purification process subsequent to expression.","Examples of such fusion expression systems are now widely used and have been commercialised by several suppliers.","Such fusion peptide sequences are attached to the amino or carboxyl terminal end of a protein sequence and are recognised by specific antibodies or affinity resins.","The expressed proteins must be solubilised from the cellular debris sometimes requiring harsh conditions including unphysiological pH values or use of chaotropic reagents and therefore the affinity purification process must be robust enough to function under such conditions.","By using this sequence as a means of isolating or purifying the expressed fusion protein, the need for additional purification tags is eliminated.","Thus this sequence has a dual purpose.","In some cases it may be desirable to use a further peptide sequence tag as a means of isolating or purifying the expressed fusion protein.","The sequence is preferably between 1 and 30 amino acids in length.","The peptide sequence tag sequence (20) may be located at the N-terminal or C-terminal region of the antigen or antibody binding protein as shown in FIG.","13.It is, however, preferably located at the opposite end of the antigen or antibody binding protein to which SEQ ID NO 1 is fused.","Where the additional peptide sequence tag is located on the same terminal region as SEQ ID NO 1, it is preferably fused to the free end of SEQ ID NO 1.Many peptide sequence tags are known in the art.","Examples of suitable peptide sequence tags for the purposes of the present invention are described in U.S. Pat.","No.","4,569,794A, and EP0 282 042B, the contents of which are herein incorporated by reference.","Preferably, the peptide sequence tag comprises at least one histidine amino acid.","Even more preferably the peptide sequence tag has the formula His-X in which X is selected from the group consisting of -Gly-, -His-, -Tyr-, -Gly-, -Trp-, -Val-, -Leu-, -Ser-, -Lys-, -Phe-, -Met-, -Ala-, -Glu-, -Ile-, -Thr-, -Asp-, -Asn-, -Gln-, -Arg-, -Cys- and -Pro-.","Alternatively the peptide sequence tag has the formula Y-His wherein Y is selected from -Gly-, -Ala-, -His- and -Tyr-.","Particularly suitable peptide sequence tags are described in EP 0 282 042B, and a preferred example is a hexa His tag.","In one embodiment of the method of the invention, step (iii) is effected using a further antibody or a binding fragment thereof, which is specific for the peptide of amino acid sequence including SEQ ID NO 1.The said further antibody may be raised using conventional techniques to the peptide (7) which includes an amino acid of SEQ ID NO 1.This method is illustrated diagrammatically in FIG.","2.The said further antibody is an anti-fusion antibody (8), which may be immobilised on a column, magnetic bead (9) or pipette tip, for example using a secondary antibody which is suitably an anti-species antibody (10) or other methods described in the literature, such as using an antibody binding protein such as Protein A, Protein G or Protein L, bound to the bead (9).","This approach is highly suited to automation and to the isolation of large numbers, but small quantities, of novel fusion proteins in parallel.","After separation from the cell lysate residue, the bound fusion protein (4) can subsequently be eluted by increasing the pH from 7.0 to 9.0.In an alternative embodiment, the fusion protein is isolated using a separation material which has some affinity for biotin but which releases the biotin fairly readily.","Suitably the separation material is a modified version of avidin or streptavidin, which has lower affinity for biotin than native avidin or streptavidin.","A particular example of such a material is a modified version of avidin marketed as CaptAvidin™ by by Molecular Probes (Eugene, Oreg., USA).","In this embodiment, the fusion protein is isolated from the cellular debris, detergents and salts etc from the culture medium, by lowering the pH of the cell lysis mixture to pH 6.0 followed by affinity purification with CaptAvidin™ attached to (a) magnetic beads or (b) pipette tips using conventional methods.","Bound fusion protein may then be eluted from the magnetic beads or mini columns by subsequently increasing the pH from 6.0 to 9.5.Prior to the step (iv), it is preferable to confirm the identity of the expressed fusion protein.","In one particular technique, a very low volume (10 μl) of the isolated fusion protein is removed from the microtitre plate.","The sample is digested by trypsin (using methods well known in the art).","The resultant peptide extract is desalted and concentrated using a ZipTip™ (Millipore, Mass., USA) or equivalent, before analysis via mass spectrometry.","Since the sequence of the fusion protein is known, identification by MALDI spectrometry to identify the peptides is usually sufficient to confirm the identity of the fusion protein.","This technique is widely used in protein research and is summarised by T. Rabilloud (Editor) Proteome Research: 2D gel electrophoresis and identification methods.","Furthermore, this technique can be automated and there are a number of commercially available systems from companies including Amersham Pharmacia Biotech, Bio Rad, AbiMed and Genomic Solutions (WO074852A1) that will perform this function.","Similarly, prior to step (iv), it is preferable that the concentration of each expressed protein should be normalised where possible to eliminate variation between elements.","Large variations in protein density cause difficulties in interpreting the data derived from such arrays (see Ekins, Clinical Chemistry (1998) 44:9 2015-2030, U.S. Pat.","No 5,807,755 and U.S. Pat.","No.","5,432,099 for a detailed discussion on the quantitative aspects of protein immunoassays and protein arrays and definitions of assay sensitivity).","Protein normalisation can be achieved by either determining the total protein concentration and or by including internal controls in the protocols.","In a particularly preferred embodiment of the method of the invention, the fusion tag is used as an internal control and is detected by an antibody with a high affinity for the peptide of amino acid sequence which includes SEQ ID NO 1 within the fusion protein.","Alternatively, the fusion protein can be expressed with a further peptide sequence tag and this can be used as an internal control.","Such a tag may be the said further peptide sequence tag such as a hexa His tag as discussed above, which is expressed as part of the biotinylated fusion protein.","The tagged version of this fusion protein may be detected through the use of an appropriate antibody such as an anti His tag antibody.","This can be done by performing a classic immunoassay sandwich simultaneously with, or during, a subsequent analysis of a biological sample using the array.","Using the antibody to the biotinylated fusion peptide, it is possible to determine the content of the fusion protein per μl and as a ratio of total protein present.","The methodology may be performed using a sheep polyclonal primary antibody and secondary antibody sandwich in which the secondary antibody is conjugated with fluorescent dye (e.g.","goat anti-mouse antibody conjugated to Alexa 488, Molecular Probes, Eugene, USA).","The fluorescent dye used is spectrally distinct from any used with the secondary antibody for the biological sample.","Both processes have been optimised for automation.","The avidin or steptavidin coated non-porous support used in step (iv) of the method of the invention is suitably a glass or plastics material.","Such supports are well suited to the production of small concentrated arrays.","This is important, since biological samples are generally very limited in volume, and thus very valuable.","A minimal surface area containing the targets is required for protein arrays, while still enabling the ability to achieve the required sensitivity of the assay is desirable.","In addition, high density of either antigen or antibody in the array produces better signal to noise ratios when used in an assay.","Furthermore, as compared to supports with porous surfaces including membranes, non-porous supports are more physically robust, are well suited to automation and have a lower background when imaged on fluorescent scanners.","These may be coated with avidin or streptavidin using conventional methods.","For example, the immobilisation of streptavidin to non-porous surfaces such as polystyrene multi-well plates is well known in the art.","In its most basic form, a solution of streptavidin is left in contact with the surface for some hours.","Un-bound protein is then removed by washing and the residual active moieties on the plastic surface blocked with BSA or an equivalent.","Although this approach may be passive, it is effective.","The non-covalent binding of streptavidin to polystyrene or nitrocellulose surfaces appears to be highly stable and resistant to elevated temperatures and high concentrations of chaotropic reagents, as described in WO98/37236.Avidin can be chemically attached to glass using the N-hydroxysuccinamide active ester of avidin as described by Manning, et al.","Biochemistry 16: 1364-1370 (1977) and can be attached to nylon via carbodiimide based coupling methods as described by Jasiewicz, et al.","Exp.","Cell Res.","100: 213-217 (1976).","In another method, high molecular weight compounds such as biotin-N-hydroxy-succinimide ester, N-biotinyl-6-aminocaproyl-N-hydroxysulfosuccinimide ester, sulfosuccinimidyl-2-(biotinamido)ethyl-1,3-dithiopropionate were biotinylated and used to coat a suitable surface.","Avidin or streptavidin was then coated in a second layer and was retained through binding the biotin linker attached to the high molecular weight compound as described in EP0620438.Attachment of streptavidin via a layer of biotin on the support surface was further developed in WO98/59243, which describes how biotin can be attached to a surface by chemical means or by light activation at 365 nm.","The benefit that this provides is that regions of the surface can be masked.","The elegance of these approaches is that biotin can be covalently bound to glass surfaces and will, in turn, non-covalently bind streptavidin only in those areas of the support that have been treated.","This enables patterns of streptavidin “acceptor” protein on the support to be manufactured if required.","In a preferred embodiment of the invention however, the entire surface of the non-porous support is coated with avidin or streptavidin, and then areas which are not required for binding are blocked, for example by addition of bovine serum albumin (BSA).","In this way, any non-specific interaction of fusion protein with the support is reduced.","Proteins that have been applied directly onto glass or plastic surfaces become non-covalently bound through interactions with charged groups on the solid surface the active moieties of the solid surface (typically silanol groups in glass or charged surface residues on polystyrene).","Such non-specific adsorption of antigens or antibodies onto the surfaces of glass and plastic significantly reduces their antigenicity and antigen binding capacity respectively.","The avidin or streptavidin layer therefore fulfils a dual role of firstly attaching the biotinylated fusion protein (non-biotinylated proteins that co-purify do not bind enabling a further purification step) and secondly, the dense layer of streptavidin shields the biotinylated fusion protein from undesirable non-specific interactions with the support surface.","When the fusion protein is applied to the avidin or streptavidin coating on the support in step (iv), very tight but non-covalent bonding occurs.","Preferably, the non-porous support is coated with streptavidin.","Biotin attachment to streptavidin is multivalent, providing a binding of very high capacity when compared to that of the antigen bound directly to the support surface.","Once the proteins have been applied to form the array, the bonding is strong enough to withstand extensive and stringent washing without appreciable loss of fusion protein.","This is illustrated in FIG.","3.Biotinylated fusion protein (4) is attached to the surface of the array support (12) via tight, non-covalent interaction with streptavidin (14).","In the preferred example, streptavidin (14) is covalently bound to the support material.","Sites on the array support material to which no streptavidin molecules are bound, are blocked by BSA or other surface modifiers (13).","Fusion proteins bind the steptavidin (14) via the biotin label (7) on the fusion peptide (6).","Furthermore, the avidin or streptavidin layer, whether attached directly to the surface of the support or via a biotinylated linker, is highly stable, is capable of being stored dry and can be heated and or treated with aggressive reagents without apparent loss of function (unlike most antigens and antibodies).","Further “acceptor” layers can be constructed on top of the foundation of the streptavidin layer if required.","These may comprise other antibody binding proteins known in the art.","The array will have the advantage of using the concentrating effect of the streptavidin, which has multivalent sites for biotin attachment.","This enables four times the biotin interaction with both antigen arrays and antibody arrays.","This allows higher densities of either antigen or antibody in each element of the array which in turn means that more elements can be assembled per mm2 while achieving the same signal (see U.S. Pat.","No.","5,807,755 and U.S. Pat.","No.","5,432,099 for a discussion of the quantitative aspects) and less surface area of the solid support is utilised.","The advantage is that less biological sample is thus required.","Where the array is to consist of antigens arrayed on a microscope format, it suitably contains a large number of these, for example from 3-10,000 different fusion proteins.","These may be generated or obtained from various sources, depending upon the intended nature and target for the analysis to be conducted using the array.","Preferably, however, each protein will be expressed in the form of two fusion proteins, one with the peptide including SEQ ID NO 1 attached to the C-terminal, and one with the peptide including SEQ ID NO 1 attached to the N-terminal.","In this way, the relative antigenicities of each version of the fusion protein to a complementary antibody can be assessed.","The uses of antibody binding proteins such as Proteins A, G and L have been extensively reported.","The binding of such proteins to antibodies is sufficiently tight to enable use in separation and detection techniques.","These proteins are known to bind to the conserved regions of various classes of antibody.","When the method of the invention is used to produce antibody arrays, antibody attachment is achieved by capture of the antibody via use of a layer of antibody binding proteins fused to a peptide which comprises SEQ ID NO 1.This layer preferably comprises of a mixture of biotinylated tagged Proteins A, G and L, and more preferably, some of which are fused at the C-terminal end, and some of which are labelled at the N-terminal end.","By creating a mixture of antibody-binding proteins, a universal acceptor is created enabling the attachment of virtually any antibody, polyclonal, monoclonal, full-chain fragments, single chain antibodies and phage with antibody activity into which a Protein A, G or L site is present or has been engineered.","Any antibody can be incorporated into the array without the need to pre-process or modify the antibody.","When such antibody binding proteins are applied to an avidin or streptavidin coated surface in step (iv) a very high density of bound protein (biotinylated tagged protein binding to multivalent streptavidin) results.","This method has the advantage that only one amino acid residue is biotinylated, and this is part of the fusion peptide, leaving the antibody binding sites on the lectins available.","These antibody binding proteins effectively create a second layer on the solid surface.","This layer comprising of bound fusion tagged Proteins A, G and L, acts as a universal acceptor surface for any antibody (FIG.","6) without the need for direct biotinylation of the antibody.","This saves time, antibody and eliminates the possible degradation of the antibody's binding to its corresponding antigen.","In a preferred embodiment, a molar excess of antibodies is pre-mixed with biotinylated peptide antibody-binding protein (e.g.","Proteins A, G or L) fusion and incubated for up to 15 minutes.","This antibody-antibody binding protein mixture is then applied directly to the streptavidin covered array support.","Alternatively, the biotinylated antibody binding protein fusion may first be applied to the streptavidin covered array support.","Individual antibodies are then applied to the surface of the coated support to form an array.","The array produced by either method comprises very discrete spots with minimal observable diffusion, leading to a good array for assay purposes.","The array obtained using the method of the invention is suitably used in methods for detecting binding between antigens and antibodies.","Thus in a second aspect, the invention provides a method of detecting binding between an antibody and an antigen, said method comprising the steps of (vi) applying to the array obtained using a method of the first aspect a sample which contains or is suspected of containing an antibody in the case of an array of step (v)(a), or an antigen in the case of the array of step (v)(b); and (vii) detecting bound antibody or antigen on the support.","Steps (vi) and (vii) of the method of the invention are suitably carried out in a conventional manner, using well known immunlogical techniques such as ELISAs, including sandwich ELISAs using labelled and in particular fluorescently labelled antibodies.","This is illustrated in the case of an antigen array in FIG.","4.Antigens (4) bound to the array substrate (12) via streptavidin (14) are detected with a suitable primary antibody (15).","The signal is amplified using a suitable secondary antibody (16) conjugated to a label (17).","The label in the preferred embodiment is a fluorescent dye, such as Alexa 488, but may be any number of other types of label that are known in the art.","Suitably the protein array continues to be monitored for quality and in particular the density of the protein during use of protein analysis devices.","This is achieved in accordance with a preferred embodiment of the invention by using the peptide which comprises SEQ ID NO 1 or the further peptide sequence tag such as the hexa His Tag mentioned above, or any other suitable tag which performs this function as an internal standard, in a manner similar to that described above for pre-array protein normalisation.","Once the antigens from numerous protein preparations have been arrayed onto the support surface, the array can then be used to assess antibody quality (see WO 99/39210) or can be used to determine antibody titre in serum samples (Joos et al Electrophoresis 2000, 21, 2641-2650).","In these instances, the relative amounts of protein between the different elements in an array can be determined by adding an internal standard to the primary sample.","The internal standard that is preferred is a sheep polyclonal antibody raised against the fusion peptide.","This is spiked into the primary antibody solution (either antibody or serum) and is detected by an anti-sheep secondary antibody conjugated with a suitable fluorescent dye that is spectrally distinct from the labelled secondary antibody to the primary sample.","A two-colour image is generated using a commercially available slide imager and the signal for each element is normalised to the signal resulting from the fusion protein.","Combined with pre-array protein content normalisation, arrays of considerable consistency can be generated.","Preferably at least some and most preferably all of the steps of the process described above are operated automatically to increase throughput and reduce labour time and costs.","Creating antigen arrays with many novel proteins means proteins must be attached with the minimum number of steps if the process is to be viable.","The use of a peptide which is readily and specifically biotinylated and which can act not only as a binding protein for assay purposes, but also as a purification means and an internal control for monitoring quality, provides just such a method.","The method of the invention allows diverse collections of proteins to be attached with universal procedures, a minimum number of steps and maximum predictability of orientation.","The method is suitable for operation on a large-scale, for example in high-throughput screening.","In a third aspect the invention provides a protein array on a non-porous support, obtained using the method of the first aspect of the invention.","Some elements used in the above-described methods are novel and therefore form further aspects of the invention.","In particular, in a fourth aspect, the invention provides a fusion protein comprising an antibody binding protein fused at the N— or C-terminus to a peptide of 13 to 50 amino acids which comprises SEQ ID NO 1, such as a peptide of SEQ ID NO 2.In particular, the antibody binding protein is Protein A, G or L and preferably a mixture thereof.","The fusion protein may additionally comprise a further peptide sequence tag such as the hexa His tag or another suitable sequence tag, which are known to those skilled in the art as discussed above.","Such sequence tag may be located at the N or the C-terminus of the antigen or antibody binding protein.","It is, however, preferably located at the opposite end of the antigen or antibody binding protein to which the amino acid sequence of SEQ ID NO 1 is fused.","Where it is located at the same terminal region as SEQ ID NO 1, the sequence tag is fused to the free end of SEQ ID NO 1.A fifth aspect of the invention comprises a nucleic acid sequence which encodes the fusion protein of the fourth aspects.","In particular in this case, the sequence which encodes the peptide is suitably of SEQ ID NO 9.","(SEQ ID NO 9) GGCCTGAACGACATCTTCGAGGCTCAGAAAATCGAATGGCACGAA DESCRIPTION OF THE FIGURES FIG.","1 illustrates diagrammatically the expression of a protein which is either an antigen or an antibody binding protein such as Protein A, G or L, in a form in which it can be used in the method of the invention.","FIG.","2 illustrates diagrammatically the isolation of fusion protein from cellular debris using an anti-tag antibody in accordance with an embodiment of the invention.","FIG.","3 illustrates diagrammatically the attachment of expressed fusion protein to a support surface coated with streptavidin in accordance with an embodiment of the invention.","FIG.","4 illustrates diagrammatically the detection of bound antigen with classic ELISA sandwich with the secondary conjugated to a fluorescent marker such as Alexa 488 in accordance with an embodiment of the invention.","FIG.","5 shows the results of a series of experiments in which a fusion protein comprising of GST fused to the fusion peptide was arrayed onto a streptavidin-coated microscope slide at several concentrations, the lowest in the above example being equivelent to 500 pg/spot.","Panel (a) shows the image produced by a commercially available scanner using an excitation wavelength of 485 nm and an emission wavelength of 520 nm.","In this example, the secondary antibody was conjugated to Alexa 488 as described in the text.","Panel (b) shows an inverted image of (a) for ease of viewing.","Pannel (c) shows an enlarged area of the array support with fusion protein at 500 pg/spot.","The signal: noise ratio for these spots indicates that detection limits (signal:noise ratios of 3:1) would give a detection limit in the order of 10-50 pg protein per feature.","FIG.","6 (a) illustrates diagrammatically how solutions of Protein A, G and L (18) expressed as both C— and N-terminal fusion proteins were incubated with the streptavidin-coated slide (12).","(a) Solutions of Proteins A, G and L (18) expressed as both C— and N-terminal fusion proteins were incubated with the streptavidin-coated slide (12).","Antibodies (19) can be attached to the slide in highly discrete spots by arraying with a solid pin device or similar.","The antibodies bind to the divalent protein binding proteins (Proteins A, G or L) at high densities.","This was exemplified by binding a goat anti-mouse Ab conjugated to Alexa 488 and the image was acquired by scanning the slide with an emission wavelength of 485 nm and an excitation wavelength of 520 nm (inset panel (b)) FIG.","7 shows the structure of the pAN-4 DNA vector obtainable from Avidity Inc. in which the S-D box (ASGGA) is shown in bold type, the start methionine codon is shown in italics and underlined, the sequence encoding the peptide of SEQ ID NO 2 is underlined, the ampicillin resistance bla is shown in bold type and the laclq is shown bold and underlined; FIG.","8 shows the structure of the pAN-5 DNA vector obtainable from Avidity Inc. with annotations similar to those used in FIG.","7; FIG.","9 shows the structure of the pAN-6 DNA vector obtainable from Avidity Inc. with annotations similar to those used in FIG.","7; FIG.","10 shows the structure of the pAC-4 DNA vector obtainable from Avidity Inc. with annotations similar to those used in FIG.","7; FIG.","11 shows the structure of the pAC-5 DNA vector obtainable from Avidity Inc. with annotations similar to those used in FIG.","7; FIG.","12 shows the structure of the pAC-6 DNA vector obtainable from Avidity Inc. with annotations similar to those used in FIG.","7.FIG.","13 illustrates diagrammatically the expression of a protein, which is either an antigen, or an antibody binding protein such as Protein A, G or L, in a form in which it can be used in the method of the invention wherein the two alternative positions of the second peptide sequence tag are shown.","DETAILED DESCRIPTION OF THE INVENTION Step 1: Cloning All genes expressed were cloned from cDNA preparations directly into each of the pAN and pAC series of vectors (Avidity Inc, USA).","These were used to express N-terminal and C-terminal fusion proteins respectively.","The fusion peptide sequence used was SEQ ID NO 2 shown above.","The insert sequences were confirmed by DNA sequencing performed on 377 (PE Corporation Inc) and MagaBase (Amersham Pharamcia Biotech) instruments using the manufacturer's methodologies.","Step 2: Expression All fusion proteins were expressed under the control of the tightly repressed Trc promoter and is IPTG-inducible.","All proteins were expressed in strain AVB100 (Avidity Inc, Colo., USA), an E. coli K12 strain [MC1061 ara D139 delta(ara-leu)7696 delta(lac)174 galU galK hsdR2(rK-mK+) mcrBl rpsL(Strr)] with a birA gene stably integrated into the chromosome.","Over expression of the BirA protein was accomplished by induction with L-arabinose.","The stably integrated birA gene does not require antibiotics to be maintained, and use of AVB100 with IPTG-inducible vectors such as pAC and pAN, vectors (Avidity Inc, USA) allowed independent control over the expressed gene of interest and the BirA levels.","Strain AVB99 (Avidity Inc) was also used and is an E. coli strain (XL1-Blue) containing a pACYC184 plasmid with an IPTG-inducible birA gene to overexpress biotin ligase (pBirAcm).","Strain AVB101 (Avidity Inc) was also used and is an E. coli B strain (hsdR, lon11, sul Al), containing a pACYC184 plasmid with an IPTG-inducible birA gene to overexpress biotin ligase (pBirAcm).","Expression of both biotin ligase and the fusion protein was induced with IPTG (1 mM).","Biotin was added at the time of induction to a concentration of 50 μM.","Step 3: Purification Biotinylated fusion proteins were isolated by two separate methods.","These methods can either be used as alternates or were combined as a two-stage process where ultra-pure preparations were required.","a) Purification Using Anti-Fusion Peptide Antibodies A partially purified mouse monoclonal antibody to the C-terminus fusion peptide was available and polyclonal antibodies to the C— and N-terminal fusion proteins were raised in rabbit.","i) In one of the methodologies, the anti C-terminal mouse monoclonal was attached directly to magnetic beads using 2.4 micron magnetic beads with a tosylated activated surface (Dynal Biotech ASA, Norway) as follows: Coating procedure.","Dynabeads M-280 Tosylactivated were resuspended by pipetting and vortexing for approximately 1 min and were immediately pipetted into the reaction tube.","Supernatant was removed from the beads using a magnet (Dynal MPC) to separate the beads from solution.","The supernatant was removed, leaving beads undisturbed.","The beads were resuspended in an ample volume of 0.I M Na-phosphate buffer pH 7.4 and mixed gently for 2 min.","After using the magnet again and pipetting off the supernatant, the washed beads were resuspended in the same volume of 0.1 M Na-phosphate buffer pH 7.4 to the required concentration.","The appropriate antibody was dialysed into 0.1 M Na-phosphate buffer pH 7.4.The amount of antibody was approximately 3 μg antibody per 1 Dynabeads (approximately 20 μg/mg) and the beads were resuspended by vortexing for 1 min.","The mixture was incubated for 16-24 h at 37° C. with slow tilt rotation.","After incubation, the magnet was used to separate the magnetic beads for 1-4 minutes and the supernatant was removed The coated beads were washed four times (twice in ×1 PBS pH 7.4 [phosphate buffered saline] with 0.1% [w/v] BSA for 5 minutes at 4° C.) once with 0.2 M Tris-HCl pH 8.5 with 0.1% (w/v) BSA for 24 hours at 20° C. or for 4 hours at 37° C. (Tris blocks free tosyl-groups) and finally once in ×1 PBS, pH 7.4 with 0.1% [w/v] BSA for 5 minutes at 4° C. The Dynabeads M-280 Tosylactivated are thereby coated with the antibody.","The cells expressing the fusion protein of interest were lysed for 15 minutes in ice-cold ×1 PBS, pH 7.4 with 1% NP-40 and protease inhibitors, after which the lysate was centrifuged at 2,000×g for 3 minutes.","The lysate was pre-cleared by incubation of the ice-old lysate (in 1.5 ml Eppendorf tubes) for 2 hours with Dynabeads pre-coated with the appropriate antibody (0.5 mg Dynabeads pr.","lysate from 1×106 cells).","The Dynabeads were washed 3 times in 1.5 ml ice-cold PBS/1% NP40 by using a Dynal Magnetic Particle Concentrator to collect the beads at the wall after each washing step.","The fusion protein-antibody magnetic bead complex was disrupted by adjusting the pH to above 9.0.Supernatant was separated from the magnetic beads with the Magnetic Particle Concentrator and assayed for total protein concentration, concentration of fusion peptide and the protein was identified by mass spectrometry using a PerSeptive Voyager MADLI (see below).","ii) In the second methodology, the antibody was attached indirectly to Dynal magnetic beads via Protein A and Protein G previously immobilised onto the surface of the bead by the manufacturer.","A mixture of Dynabeads-Protein G and Dynabeads-Protein A were resupended by vortexing for 1-2 minutes.","The supernatant was removed from the beads using a magnetic workstation as described above.","0.5 ml 0.1 M Na-phosphate buffer pH 7.0 containing 0.01% Tween 20 and 0.1% (w/v) BSA were added and the wash procedure repeated three times.","The antibody was added to the washed Dynabeads and incubated with gentle mixing for 10-40 minutes.","The supernatant was removed using the magnetic workstation.","The beads were twice resuspended in 0.5 ml 0.1 M Na-phosphate buffer pH 7.0 containing 0.01% Tween 20 and 0.1% (w/v) BSA for protein stability.","Supernatant was removed and the beads added to the lysate mixture as prepared above.","Binding of the fusion protein was performed at 2-8° C. for 10 minutes to 1 hour.","Approximately 25 μg target protein per μl of the initial Dynabeads Protein G volume was used to assure an excess of protein.","Incubation was performed while tilting and rotating the tube with incubation times as low as 10 minutes.","Supernatant containing detergents and cell lysate was removed from the fusion-protein-Ig Dynabeads-Protein G complex using the magnetic workstation and washed 3 times using ×1 PBS, pH 7.4 with 0.01% Tween 20.Bound fusion protein was best eluted from the fusion-protein-Ig Dynabeads-Protein G/A complexes by adjusting the pH to above 9.0 and removing the supernatant containing the now purified fusion protein.","Supernatant was separated from the magnetic beads with the Magnetic Particle Concentrator and assayed for total protein concentration, concentration of fusion peptide and the protein was identified by mass spectrometry using a PerSeptive Voyager MADLI (see below).","iii) In another example, Proteins A, G and L mixtures were immobilised on to suitably prepared pipette tips.","The antibody was incubated with the pipette tips in 50 mM Tris-HCl buffer, pH 8.0 containing 0.01% Tween 20 and 0.1% (w/v) BSA for 60 minutes at room temperature.","The coated pipette tips were then rinsed with 3 pipette volumes of 50 mM Tris-HCl buffer, pH 8.0 containing 0.01% Tween 20 and 0.1% (w/v) BSA.","200μl cell lysate was aspirated from the bottom of the tip either by hand or with a robotic workstation several times to ensure the extraction of the biotinylated fusion protein.","The cell lysate was discarded.","The pipette tips were rinsed with three volumes of 10 mM Tris-HCl buffer, pH 8.0 containing 0.01% Tween 20 and 0.1% (w/v) BSA.","Bound fusion protein was eluted in half a pipette volume of 50 mM sodium bicarbonate-HCl buffer, pH 10.0 containing 0.01% Tween 20 by gently aspirating this aliquot up through the bottom of the pipette tip.","The resulting solution containing the fusion protein was assayed as described below.","iv) In another preferred methodology an alternative version of the biotinylated fusion protein was constructed with the addition of a hexa His tag.","The hexa His fusion peptide is often used as a standard purification procedure and is well known to those skilled in the art.","Typically, cells were lysed in 5 ml buffer per gram wet weight of cells.","The lysis buffer comprised: ×1 NBB (20 mM Tris CL, 100 mM NaCl, 5 mM Imidazol, pH 8.0) with 1 in 100 volume of 10 mg/ml lysozyme, 1 in 100 volume protease inhibitor cocktail (Calbiochem protease inhibitopr cocktail set 3), 10 mM beta mercaptoethanol, supplemented with a ×1 detergent cocktail supplied by Novagen (Madison, USA).","The cells were lysed for 15 minutes at 30-37° C. Cellular proteins were denatured by adding Urea to a final concentration of 6M and 2M thiourea The solution was clarified by passing through a 0.22 micron filter, and then applied directly onto nickel agarose matrix (NTA supplied by Qiagen, Germany).","Proteins were incubated with the nickel agarose beads for 15 minutes and the non-binding protein removed by centrifugation.","The beads were washed three times in 10 volumes of the lysis buffer supplemented with 6M Urea and 2M Urea.","After the final wash, 50% of the wash buffer was removed and then diluted with a 20 mM Tris HCl, 100 mM NaCl, pH 8.0 buffer containing 10 mM beta mercaptoethanol.","This step was repeated three times.","Finally the beads were washed with 10 volumes of buffer, the composition of which was 20 mM Tris HCl, 100 mM NaCl, pH 8.0 buffer (without urea/thiourea).","The proteins were eluted several aliquots of buffer (20 mM Tris HCl, 100 mM NaCl, pH 8.0 buffer), supplemented with various concentrations of imidazole.","The typical concentration range of imidazole used to eluted the bound protein was between 20 mM to 500 mM.","The fractions containing the eluted protein were pooled.","b) Purification Using CaptAvidin™ (Molecular Probes Inc. Oregon.","USA) In another experiment, the biotinylated fusion protein was isolated using a novel form of streptavidin marketed as CaptAvidin™ (Molecular Probes, Oregon, USA) immobilised to a suitable surface.","In this modified form of streptavidin, the tyrosine residue in the biotin binding sites is nitrated, thereby reducing the very strong non-covalent bond with a Ka of 1015M−1 to a Ka of 109M−1.The association between biotin and CaptAvidin™ can therefore be disrupted by raising the pH to between 9-10 as described below: i) In one preferred embodiment, CaptAvidin™ protein was attached to tosylated magnetic beads (Dynal Biotech ASA, Norway) and was washed and prepared as described above.","The CaptAvidin™ coated beads were washed three times in 50 mM citrate phoasphate buffer, pH 4.0 containing 0.01% Tween 20 and 0.1% (w/v) BSA and the supernatant was discarded.","The cell lysate mixture was prepared as described above and the pH adjusted to 5.0.CaptAvidin™ coated beads were added at a ratio of 0.5 mg Dynabeads per lysate from 1×106 cells.","The solution was incubated with gentle agitation for 10-60 minutes.","The supernatant was removed from the magnetic beads using a magnetic workstation (Dyanl Biotech ASA, Norway) and washed with three aliquots of 10 mM Tris-HCL buffer, pH 8.0 containing 0.01% Tween 20, discarding the supernatant.","The biotinylated fusion protein is detached from the CaptAvidin™ coated magnetic beads by adding an aliquot of 50 mM sodium bicarbonate-HCl buffer, pH 10.0 containing 0.01% Tween 20 and gently agitating the slurry for 15 minutes at room temperature.","The magnetic beads were removed using the magnetic workstation and the supernatant containing the biotinylated fusion protein was retained.","ii) In another example, the magnetic beads were replaced by creating mini columns of CaptAvidin™ conjugated to agarose beads (Molecular Probes Inc, Oregon, USA) mixed with an equal volume of Sepharose® CL-4B agarose (Amersham Pharmacia Biotech Ltd, UK) to increase the bed volume with mini columns made by pouring the slurry into pipette tips in 50 mM citrate phosphate buffer, pH 4.0 containing 0.01% Tween 20.Biotinylated fusion protein was separated from cell lysate mixture by affinity chromatography.","Unbound material is eluted from the column with 10 column volumes of 10 mM Tris-HCl buffer, pH 8.0 containing 0.01% Tween 20.Biotinylated fusion protein was eluted from the column in two column volumes of 50 mM sodium bicarbonate-HCl buffer, pH 10.0 containing 0.01% Tween 20.iii) In yet another experiment, the CaptAvidin™ agarose beads were immobilised into a pipette tip and fusion protein binding and elution was performed as described above.","Step 4: Protein Identification Expressed and purified fusion proteins were identified by peptide finger printing.","Using methods as reviewed in Proteome Research (Edited by Rabilloud), the fusion protein was digested with trypsin, the resulting peptide solution was desalted and concentrated using a ZipTip™ (Millipore, Mass., USA) reverse phase column, diluted into matrix solution and applied to a target plated from a PerSeptive Voyager™ mass spectrometer and analysed by MADLI.","The resulting spectra of peptide masses were compared with the anticipated peptide finger print for the protein using the ExPASy search algorithms (GeneBio AG, Switzerland) via their website (www.expasy.com).","Step 5: Protein Assay (Normalisation) A 3-5 μl aliquot of the purified fusion protein was removed from the stock solution and assayed for total protein content using the BCA method in preference to Bradford assay due to the presence of detergents in the protein samples.","The concentration of biotinylated fusion protein was determined by immunoassay as follows; A 3-5 μl aliquot of the purified fusion protein was removed from the stock solution and incubated in a black, streptavidin-coated microtitre plate (Beckton Dickenson, USA).","The well was washed three times with 50 mM Tris-HCL buffer, pH 8.0 containing 0.01% Tween 20.The well was blocked using 1% (w/v) BSA in the same buffer for 30 minutes and then rinsed three times with 50 mM Tris-HCL buffer, pH 8.0 containing 0.01% Tween 20.The immobilised biotinylated fusion protein was incubated with either an anti N-terminal or anti C-terminal polyclonal antibody raised in rabbit diluted into 50 mM Tris-HCL buffer, pH 8.0 containing 0.01% Tween 20 and 0.1% (w/v) BSA.","The well was rinsed three times with buffer and then probed with a anti-rabbit, mouse monoclonal conjugated to Alexa 488 (Molecular Probes Inc, Oregon, USA) and the signal measured with a PerkinElmer Flight fluorescence plate reader.","A standard curve with known amounts of Glutathione S-transferase expressed using the expression system described in U.S. Pat.","No.","5,723,584, U.S. Pat.","No.","5,874,239 and U.S. Pat.","No.","5,932,433 was used for calibration in the range of 0.1-500 μg of fusion protein per well.","Step 6: Manufacture of Protein Arrays a) Creation of Streptavidin Coated Microscope Slides Microscope slides coated with streptavidin were first imaged on a variety of commercially available slide readers using an excitation wavelength of 480 nm and and emission wavelength of 520 nm to assess the evenness of the coating.","b) Manufacture of Antigen Arrays The streptavidin coated slides were rehydrated with ×1 phosphate buffered saline at pH 7.3.Purified biotinylated fusion proteins at a concentration of approximately 1 μg/μl were spotted onto the surface of the slide using a solid pin with a tip diameter of 100-150 microns (Biorobotics, Cambridge, UK) by hand and with a robotic system.","The slide was incubated at room temperature in a humidity-controlled environment for 30 minutes.","The slide was then typically washed with ×1 PBS, pH 7.3 containing 0.01% (v/v) Tween and then blocked by incubating the slide with 1% (w/v) BSA for 10 minutes.","The slide was rinsed with ×1 PBS, pH 7.3 containing 0.01% (w/v) Tween 20 and then incubated with the primary antibody of choice diluted 1:400 in ×1 PBS, pH 7.3 containing 0.01% (w/v) Tween 20 and 0.1% (w/v) BSA, or a complex biological mixture of proteins containing immunoglobulins, e.g.","diluted serum samples.","The slide was then rinsed in ×1 PBS, pH 7.3 containing 0.01% (w/v) Tween and 0.1% (w/v) BSA and incubated with an appropriate secondary (for example mouse anti-human IgG monoclonal conjugated to Alexa 488 (Molecular Probes Inc) for the detection of immunoglobulins in serum, for example).","The slides were then imaged at excitation/emission wavelengths of 480/520 nm, for the Alexa 488 conjugate, although one skilled in the art can appreciate that many such secondary Abs with a variety of labels (colorimetric, alternative fluorescent, radiolabelled or chemiluminescent) could be used in its place.","An example of the results obtained is illustrated in FIG.","5 hereinafter.","c) Manufacture of Antibody Arrays Creation of a Universal Antibody Acceptor Layer Proteins A, G and L from Streptococcus aureus were cloned into the expression vectors pAN-4, pAN-5 or pAN-6, pAC4, pAC-5 and pAC-6) and were expressed and purified as described above, resulting in both C— and N-terminal fusion proteins which were biotinylated in vivo, again as described above.","Streptavidin coated microscope slides were coated with a mixture of fusion proteins (both C— and N-terminal fusions) of Proteins A, G and L in ×1 PBS, pH 7.3 at a concentration of 1 mg/ml.","The slides were incubated at room temperature for a minimum of 30 minutes in a humidity-controlled environment.","The slides were washed with ×1 PBS, pH 7.3 containing 2 mM Sodium Azide and were stored in sealed containers in a moist atmosphere (to prevent drying) at 4° C. until required.","Printing Antibody Arrays The universal antibody acceptor layer was used to attach a variety of different classes of antibodies and those phage molecules engineered to include a Protein A, G or L binding site.","Antibody preparations are diluted in 1× PBS, pH 7.3 containing 0.01% Tween to a concentration of 0.2-10 mg/ml.","The antibody solutions were applied to the universal antibody acceptor layer with solid pins with a tip diameter of between 100-150 microns (Biorobotics, Cambridge, UK) by hand or with a robotic system.","The slides were then blocked with 1% BSA in xl PBS, pH 7.3 containing 0.01% Tween.","Slides were rinsed with the ×1 PBS, pH 7.3 containing 0.01% Tween and 2 mM Sodium Azide and were stored in sealed containers in a moist atmosphere (to prevent drying) at 4° C. until required.","Scanning as described above for antigen arrays produced the sort of results which are illustrated in FIG.","6.Step 7: Labelling Complex Mixtures of Proteins with Fluorescent Dyes Typically, protein samples were prepared by solubilising them in a variety of buffers and detergents, depending on the biological sample.","Many samples required aggressive solubilisation procedures requiring the use of non-ionic detergents and 8M urea, similar to those used in the preparation of proteins for the first dimension of 2D electrophoresis gels.","For example, the solublization methodology involved homogenization of the sample into solution containing 4% CHAPS, 50 mM PBS, pH 7.6 with either 7 M urea and 2 M thiourea or 8 M urea Buffers containing primary amino groups such as TRIS and glycine inhibit the conjugation reaction and were therefore avoided.","The presence of low concentrations (<2%) of biocides such as azide or thimerosal did not affect protein labelling.","The solubilised protein was centrifuged at 10,000 g to remove cellular debris and non-solubilised material and the mixture was immediately labeled.","Complex mixtures of proteins from biological samples were labelled with a fluorescent tag prior to incubation with the antibody array as prepared above.","Clearly, those skilled in the art will recognise that other forms of labels can be applied to the technique such as radiolabelling, chemiluminescent and visual dyes.","Further, other fluorecent dyes can also be applied to the process.","One preferred embodiment is the use of Cy3 and Cy5 mono reactive dyes (Amersham Pharmacia Biotech Ltd, UK).","Dye labelling of complex protein mixtures was unpredictable and had to be optimised for each type of biological sample.","Specifically, the binding of dye molecules to proteins via residues with amine groups often reduced the antigenicity of certain proteins such that they were no longer recognised by a functional antibody.","The manufacturer's recommended procedure is designed to label 1 mg protein to a final molar dye/protein (D/P) ratio between 4 and 12.This assumes an average protein molecular weight of 155,000 daltons.","In the present invention, an average dye/protein ratio above 2-3 was found to interfere with the antibody-antigen reaction for many of the proteins studied.","It was determined that the D/P ratios could be simply controlled by using different concentrations of protein and different buffer pH values.","Altering the protein concentration and reaction pH changed the labelling efficiency of the reaction significantly.","Optimal labelling occured at pH 9 and by reducing the pH to 7.6 reduced the dye/protein ratio to between 1-3.Higher protein concentrations increased labeling and so the control of protein concentration was also found to be critical.","Solutions of up to 10 μg/μl of a single protein species gave dye/protein ratios of 10-14, so more appropriate concentrations were found to be 0.1-1.0 μg/μl.","A typical method was as follows: complex protein mixtures prepared as described above, were diluted to several concentrations in ×1 PBS buffer, pH 7.6 containing 0.2% CHAPS to achieve an average protein species concentration of 1.0 μg/μl (total protein concentration was in the range of 50-100μg/μl) The protein solution was incubated at room temperature for 30 minutes with constant gentle agitation.","Labeled protein must be separated from the excess, unconjugated dye prior to incubation with the antibody arrays.","The manufacturer recommends separation from unbound protein by gel permeation, however, due to the presence of membrane-bound proteins with poor solubility this step was replaced by simply adding an excess of glycine to the solution to halt the reaction.","The labeled protein solution was incubated for a further 15 minutes to ensure the removal of residual free dye.","Labeled proteins were stored at 2-8° C. without further manipulation.","Free dye was also removed using the method of Ünlü et al (1997) in which free dye was removed by overnight incubation with SM-2 beads (Bio-Rad, CA, USA).","The final dye/protein (D/P) ratio was estimated as follows: a portion of the labeled protein solution was diluted so that the maximum absorbance was 0.5 to 1.5 AU.","Molar concentrations of dye and protein were calculated.","The extinction coefficient will vary for different proteins but is a reasonable average to use for complex mixtures.","The ratio of the average number of dye molecules coupled to each protein molecule was calculated as follows: Cy5/Protein ratios were calculated using molar extinction coefficients of 250,000 M− 1 cm−1 at 650 nm for Cy5, and 170,000 N−1 cm−1 at 280 nm for the protein mixture.","The calculation was corrected for the absorbance of the Cy5 dye at 280 nm (approximately 5% of the absorbance at 650 nm) as per the manufacturer's product data sheets.","[Cy5 dye]=(A650)/250000, [protein]=[A280−(0.05×A650)]/170000, (D/P) final=[dye]/[protein], (D/P) final=[0.68× (A650)]/[A 280−(0.05×A650)).","Cy3 /Protein ratios were calculated using molar extinction coefficients of 150,000 M−1 1 cm−1 at 552 nm for the Cy3 dye and 170000 M−1 cm−1 at 280 nm for the protein are used in this example.","The calculation was corrected for the absorbance of the dye at 280 nm (approximately 8% of the absorbance at 552 nm).","[Cy3 dye]=(A552)/150000, [antibody]=[A 280−(0.08×AS52)]/170000, (D/P) final=[dye]/[antibody], (D/P) final=[1.13×(A552)]/[A280−(0.08×A552)).","Step 8: Determination of Protein Expression Using Antibody Arrays Cy3-labelled and Cy5-labelled proteins were mixed in equimolar amounts based on the Dye/protein ratios determined above.","100 μl of the mixture was incubated with a antibody array that had previously been rinsed with several slide volumes of ×1 PBS, pH 7.6 containing 0.01% Tween.","The labelled protein mixture was incubated at 30° C. for one hour in an automated slide processor subject to UK Patent Application GB 0028647.6 (unpublished).","The slide was then rinsed with 10 slide volumes of ×1 PBS, pH 7.6 containing 0.01% Tween.","The slides were dried by centrifugation and imaged immediately on a commercially available slide imager using the manufacturer's operating procedures.","The Cy3 and Cy5 labelled protein ratios were analysed and normalised to a number of marker proteins such as actin and GAPDH.","While this approach is suitable for similarly prepared tissues or other biological samples, care must be taken on the applicability of this normalisation strategy between different tissue types and other biological samples, since the total cell content of all proteins vary considerably from tissue to tissue.","The potential of protein arrays has been discussed for many years and clearly is a much needed tool.","The problems with expressing, purifying, assaying and in particular, attaching proteins to solid, non-porous surfaces have all proved difficult problems to solve.","Through the novel exploitation of the vector technology described in patents U.S. Pat.","No.","5,723,584, U.S. Pat.","No.","5,874,239 and U.S. Pat.","No.","5,932,433, the present invention provides a method for the preparation of both antigen and antibody arrays that allow researchers to now apply these techniques with greater success.","All references mentioned in the above specification are herein incorporated by reference.","Other modifications of the present invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention.","Although the invention has been described in connection with the specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments.","Indeed, various modifications of the described modes for carrying out the invention, which are obvious to those skilled in the art, are intended to be within the scope of the following claims."]],"string":"[\n [\n \"Protein analysis\",\n \"A method of forming an array of proteins selected from antigens or antibodies; said method comprising the steps of (i) expressing in a recombinant cell, a fusion protein which comprises either (a) an antigen or (b) an antibody binding protein, fused to a peptide having up to 50 amino acids, which peptide comprises amino acid sequence of SEQ ID NO 1 LX1X2IX3X4X5X6KX7X8X9X10 (SEQ ID NO 1) where X1 is a naturally occurring amino acid, X2 is any naturally occurring amino acid other than leucine, valine, isoleucine, tryptophan, phenylalanine or tyrosine, X3 is phenylalanine or leucine, X4 is glutamine or asparagine, X5 is alanine, glycine, serine or threonine, X6 is glycine or methionine, X7 is isoleucine, methionine or valine, X8 is glutamine, leucine, valine, tyrosine or isoleucine, X9 is tryptophan, tyrosine, valine, phenylalanine, leucine or isoleucine and X10 is any naturally occurring amino acid other than asparagine or glutamine; where said peptide is capable of being biotinylated by a biotin ligase at the lysine residue adjacent to X6; (ii) biotinylating said peptide of the fusion protein at the lysine residue adjacent X6; (iii) isolating the biotinylated fusion protein; (iv) applying the biotinylated fusion protein to an avidin or streptavidin coated non-porous support; (v) forming an array of at least three different proteins on the support by either (a) where the fusion protein comprises an antigen, carrying out steps (i) to (iv) the desired number of times to form an antigen array; or (b) where the fusion protein comprises an antibody binding protein, applying to said protein, either prior to or after step (iv) a plurality of different antibodies or binding fragments thereof.\"\n ],\n [\n \"1.A method of forming an array of proteins selected from antigens or antibodies; said method comprising the steps of (i) expressing in a recombinant cell, a fusion protein which comprises either (a) an antigen or (b) an antibody binding protein, fused to a peptide having up to 50 amino acids, which peptide comprises an amino acid sequence of SEQ ID NO:1 LX1X2IX3X4X5X6KX7X8X9X10 (SEQ ID NO: 1) where X1 is a naturally occurring amino acid, X2 is any naturally occurring amino acid other than leucine, valine, isoleucine, tryptophan, phenylalanine or tyrosine, X3 is phenylalanine or leucine, X4 is glutamic acid or aspartic acid, X5 is alanine, glycine, serine or threonine, X6 is glutamine or methionine, X7 is isoleucine, methionine or valine, X8 is glutamic acid, leucine, valine, tyrosine or isoleucine, X9 is tryptophan, tyrosine, valine, phenylalanine, leucine and isoleucine and X10 is any naturally occurring amino acid other than aspartic acid or glutamic acid; where said peptide is capable of being biotinylated by a biotin ligase at the lysine residue adjacent to X6; (ii) biotinylating said peptide of the fusion protein at the lysine residue adjacent X6; (iii) isolating the biotinylated fusion protein; (iv) applying the biotinylated fusion protein to an avidin or streptavidin coated non-porous support; (v) forming an array of at least three different proteins on the support by either (a) where the fusion protein comprises an antigen, carrying out steps (i) to (iv) the desired number of times to form an antigen array; or (b) where the fusion protein comprises an antibody binding protein, applying to said protein, either prior to or after step (iv), a plurality of different antibodies or binding fragments thereof.\",\n \"2.A method according to claim 1 wherein the peptide of SEQ ID NO:1 is selected from Leu His His Ile Leu Asp Ala Gln (SEQ ID NO; 30) Lys Met Val Trp Asn His Arg; Leu Asn Ala Ile Phe Glu Ala Met (SEQ ID NO: 33) Lys Met glu Tyr Ser Gly; Leu Gly Gly Ile Phe Glu Ala Met (SEQ ID NO: 34) Lys Met Glu Leu Arg Asp; Leu Ser Asp Ile Phe Glu Ala Met (SEQ ID NO: 37) Lys Met Val Tyr Arg Pro Cys; Leu Ser Asp Ile Phe Asp Ala Met (SEQ ID NO: 39) Lys Met Val Tyr Arg Pro Gln; Leu Lys Gly Ile Phe Glu Ala Met (SEQ ID NO: 41) Lys Met Glu Tyr Thr Ala Met; Leu Glu Gly Ile Phe Glu Ala Met (SEQ ID NO: 42) Lys Met Glu Tyr Ser Asn Ser; Leu Lys Glu Ile Phe Glu Gly Met (SEQ ID NO: 47) Lys Met Glu Phe Val Lys Pro; Arg Pro Val Leu Glu Asn Ile Phe (SEQ ID NO: 50) Glu Ala Met Lys Met Glu Val Trp Lys Pro; Thr Arg Ala Leu Leu Glu Ile Phe (SEQ ID NO: 57) Asp Ala Gln Lys Met Leu Tyr Gln His Leu; Met Ala Ser Ser Leu Arg Gln Ile (SEQ ID NO: 73) Leu Asp Ser Gln Lys Met Glu Trp Arg Ser Asn Ala Gly Gly Ser; Met Ala His Ser Leu Val Pro Ile (SEQ ID NO: 75) Phe Asp Ala Gln Lys Ile Glu Trp Art Asp Pro Phe Gly Gly Ser; Met Gly Pro Asp Leu Val Asn Ile (SEQ ID NO: 76) Phe Glu Ala Gln Lys Ile Glu Trp His Pro Leu Thr Gly Gly Ser; Met Ala Phe Ser Leu Arg Ser Ile (SEQ ID NO: 77) Leu Glu Ala Gln Lys Met Glu Leu Arg Asn Thr Pro Gly Gly Ser; Met Ala Gly Gly Leu Asn Asp Ile (SEQ ID NO: 78) Phe Glu Ala Gln Lys Ile Glu Trp His Glu Asp Thr Gly Gly Ser; Met Ser Ser Tyr Leu Ala Pro Ile (SEQ ID NO: 79) Phe Glu Ala Gln Lys Ile Glu Trp His Ser Ala Tyr Gly Gly Ser; Met Ala Lys Ala Leu Gln Lys Ile (SEQ ID NO: 80) Leu Glu Ala Gln Lys Met Glu Trp Arg Ser His Pro Gly Gly Ser; Met Ala Gly Ser Leu Ser Thr Ile (SEQ ID NO: 82) Phe Asp Ala Gln Lys Ile Glu Trp His Val Gly Lys Gly Gly Ser; Met Ala Gln Gln Leu Pro Asp Ile (SEQ ID NO: 83) Phe Asp Ala Gln Lys Ile Glu Trp Arg Ile Ala Gly Gly Gly Ser; Met Ala Gln Arg Leu Phe His Ile (SEQ ID NO: 84) Leu Asp Ala Gln Lys Ile Glu Trp His Gly Pro Lys Gly Gly Ser; Met Ala Gly Cys Leu Gly Pro Ile (SEQ ID NO: 85) Phe Glu Ala Gln Lys Met Glu Trp Arg His Phe Val Gly Gly Ser; Met Ala Trp Ser Leu Lys Pro Ile (SEQ ID NO: 86) Phe Asp Ala Gln Lys Ile Glu Trp His Ser Pro Gly Gly Gly Ser; Met Ala Leu Gly Leu Thr Arg Ile (SEQ ID NO: 87) Leu Asp Ala Gln Lys Ile Glu Trp His Arg Asp Ser Gly Gly Ser; and Met Ala gly Ser Leu Arg Gln Ile (SEQ ID NO: 88) Leu Asp Ala Gln Lys Ile Glu Trp Arg Arg Pro Leu Gly Gly Ser.\",\n \"3.A method of forming an array of proteins selected from antigens or antibodies; said method comprising the steps of (i) expressing in a recombinant cell, a fusion protein which comprises either (a) an antigen or (b) an antibody binding protein, fused to a peptide having up to 50 amino acids or a fragment thereof having at least 13 amino acids, which peptide comprises the sequence selected from Leu Glu Glu Val Asp Ser Thr Ser (SEQ ID NO: 14) Ser Ala Ile Phe Asp Ala Met Lys Met Val Trp Ile Ser Pro Thr Glu Phe Arg; Gln Gly Asp Arg Asp Glu Thr Leu (SEQ ID NO: 15) Pro Met Ile Leu Arg Ala Met Lys Met Glu Val Tyr Asn Pro Gly Gly His Glu Lys; Ser Lys Cys Ser Tyr Ser His Asp (SEQ ID NO: 16) Leu Lys Ile Phe Glu Ala Gln Lys Met Leu Val His Ser Tyr Leu Arg Val Met Tyr Asn Tyr; Met Ala Ser Ser Asp Asp Gly Leu (SEQ ID NO: 17) Leu Thr Ile Phe Asp Ala Thr Lys Met Met Phe Ile Arg Thr; Ser Tyr Met Asp Arg Thr Asp Val (SEQ ID NO: 18) Pro Thr Ile Leu Glu Ala Met Lys Met Glu Leu His Thr Thr Pro Trp Ala Cys Arg; Ser Phe Pro Pro Ser Leu Pro Asp (SEQ ID NO: 19) Lys Asn Ile Phe Glu Ala Met Lys Met Tyr Val Ile Thr; Ser Val Val Pro Glu Pro Gly Trp (SEQ ID NO: 20) Asp Gly Pro Phe Glu Ser Met Lys Met Val Tyr His Ser Gly Ala Gln Ser Gly Gln; Val Arg His Leu Pro Pro Pro Leu (SEQ ID NO: 21) Pro Ala Leu Phe Asp Ala Met Lys Met Glu Phe Val Thr Ser Val Gln Phe; Asp Met Thr Met Pro Thr Gly Met (SEQ ID NO: 22) Thr Lys Ile Phe Glu Ala Met Lys Met Glu Val Ser Thr; Ala Thr Ala Gly Pro Leu His Glu (SEQ ID NO: 23) Pro Asp Ile Phe Leu Ala Met Lys Met Glu Val Val Asp Val Thr Asn Lys Ala Gly Gln; Ser Met Trp Glu Thr Leu Asn Ala (SEQ ID NO: 24) Gln Lys Thr Val Leu Leu; Ser His Pro Ser Gln Leu Met Thr (SEQ ID NO: 25) Asn Asp Ile Phe Glu Gly Met Lys Met Leu Tyr His; Thr Ser Glu Leu Ser Lys Leu Asp (SEQ ID NO: 27) Ala Thr Ile Phe Ala Ala Met Lys Met Gln Trp Trp Asn Pro Gly; Val Met Glu Thr Gly Leu Asp Leu (SEQ ID NO: 28) Arg Pro Ile Leu Thr Gly Met Lys Met Asp Trp Ile Pro Lys; Pro Gln Gly Ile Phe Glu Ala Gln (SEQ ID NO: 31) Lys Met Leu Trp Arg Ser; Leu Ala Gly Thr Phe Glu Ala Leu (SEQ ID NO: 32) Lys Met Ala Trp His Glu His; Leu Leu Arg Thr Phe Glu Ala Met (SEQ ID NO: 35) Lys Met Asp Trp Arg Asn Gly; Leu Ser Thr Ile Met Glu Gly Met (SEQ ID NO: 36) Lys Met Tyr Ile Gln Arg Ser; Leu Glu Ser Met Leu Glu Ala Met (SEQ ID NO: 38) Lys Met Gln Trp Asn Pro Gln; Leu Ala Pro Phe Phe Glu Ser Met (SEQ ID NO: 40) Lys Met Val Trp Arg Glu His; Leu Leu Gln Thr Phe Asp Ala Met (SEQ ID NO: 43) Lys Met Glu Trp Leu Pro Lys; Val Phe Asp Ile Leu Glu Ala Gln (SEQ ID NO: 44) Lys Val Val Thr Leu Arg Phe; Leu Val Ser Met Phe Asp Gly Met (SEQ ID NO: 45) Lys Met Glu Trp Lys Thr Leu; Leu Glu Pro Ile Phe Glu Ala Met (SEQ ID NO: 46) Lys Met Asp Trp Arg Leu Glu; Leu Gly Gly Ile Glu Ala Gln Lys (SEQ ID NO: 48) Met Leu Leu Tyr Arg Gly Asn; Arg Ser Pro Ile Ala Glu Ile Phe (SEQ ID NO: 51) Glu Ala Met Lys Met Glu Tyr Arg Glu Thr; Gln Asp Ser Ile Met Pro Ile Phe (SEQ ID NO: 52) Glu Ala Met Lys Met Ser Trp His Val Asn; Asp Gly Val Leu Phe Pro Ile Phe (SEQ ID NO: 53) Glu Ala Met Lys Met Ile Arg Leu Glu Thr; Val Ser Arg Thr Met Thr Asn Phe (SEQ ID NO: 54) Glu Ala Met Lys Met Ile Tyr His Asp Leu; Asp Val Leu Leu Pro Thr Val Phe (SEQ ID NO: 55) Glu Ala Met Lys Met Tyr Ile Thr Lys; Pro Asn Asp Leu Glu Arg Ile Phe (SEQ ID NO: 56) Asp Ala Met Lys Ile Val Thr Val His Ser; Arg Asp Val His Val Gly Ile Phe (SEQ ID NO: 58) Glu Ala Met Lys Met Tyr Thr Val Glu Thr; Gly Asp Lys Leu Thr Glu Ile Phe (SEQ ID NO: 59) Glu Ala Met Lys Ile Gln Trp Thr Ser Gly; Leu Glu Gly Leu Arg Ala Val Phe (SEQ ID NO: 60) Glu Ser Met Lys Met Glu Leu Ala Asp Glu; Val Ala Asp Ser His Asp Thr Phe (SEQ ID NO: 61) Ala Ala Met Lys Met Val Trp Leu Asp Thr; Gly Leu Pro Leu Gln Asp Ile Leu (SEQ ID NO: 62) Glu Ser Met Lys Ile Val Met Thr Ser Gly; Arg Val Pro Leu Glu Ala Ile Phe (SEQ ID NO: 63) Glu Gly Ala Lys Met Ile Trp Val Pro Asn Asn; Pro Met Ile Ser His Lys Asn Phe (SEQ ID NO: 64) Glu Ala Met lys Met Lys Phe Val Pro Glu; Lys Leu Gly Leu Pro Ala Met Phe (SEQ ID NO: 65) Glu Ala Met Lys Met Glu Trp His Pro Ser; Gln Pro Ser Leu Leu Ser Ile Phe (SEQ ID NO: 66) Glu Ala Met Lys Met Gln Ala Ser Leu Met; Leu Leu Glu Leu Arg Ser Asn Phe (SEQ ID NO: 67) Glu Ala Met Lys Met Glu Trp Gln Ile Ser; Asp Glu Glu Leu Asn Gln Ile Phe (SEQ ID NO: 68) Glu Ala Met Lys Met Tyr Pro Leu Val His Val Thr Lys; Ser Asn Leu Val Ser Leu Leu His (SEQ ID NO: 70) Ser Gln Lys Ile Leu Trp Thr Asp Pro Gln Ser Phe Gly; Leu Phe Leu His Asp Phe Leu Asn (SEQ ID NO: 71) Ala Gln Lys Val Glu Leu Try Pro Val Thr Ser Ser Gly; Ser Asp Ile Asn Ala Leu Leu Ser (SEQ ID NO: 72) Thr Gln Lys Ile Tyr Trp Ala His; Met Ala Phe Gln Leu Cys Lys Ile (SEQ ID NO: 81) Phe Try Ala Gln Lys Met Clu Trp His Gly Val Gly Gly Gly Ser, and; Met Ala Asp Arg Leu Ala Tyr Ile (SEQ ID NO: 89) Leu Glu Ala Gln Lys Met Glu Trp His Pro His Lys Gly Gly Ser, where said peptide is capable of being biotinylated by a biotin ligase; (ii) biotinylating said peptide of the fusion protein; (iii) isolating the biotinylated fusion protein; (iv) applying the biotinylated fusion protein to an avidin or streptavidin coated non-porous support; (v) forming an array of at least three different proteins on the support by either (a) where the fusion protein comprises an antigen, carrying out steps (i) to (iv) the desired number of times to form an antigen array; or (b) where the fusion protein comprises an antibody binding protein, applying to said protein, either prior to or after step (iv), a plurality of different antibodies or binding fragments thereof.\",\n \"4.A method according to claim 1 wherein the fusion protein further comprises a second peptide sequence capable of acting as an affinity or detection tag sequence to the fusion protein wherein the sequence comprises between 1 and 30 amino acids.\",\n \"5.A method according to claim 4 wherein the second peptide sequence is fused to the end of the amino acid sequence of SEQ ID NO:1 or a peptide of 13-50 amino acids comprising a sequence selected from SEQ ID NOS: 14-25, 27, 28, 31, 32, 35, 36, 38, 40, 43-46, 48, 51-56, 58-68, 70-72, 81 and 89.6.A method according to claim 4 wherein the second peptide sequence is fused to the opposite end of the antigen or antibody binding protein to which the amino acid sequence of SEQ ID NO: 1 or a peptide of 13-50 amino acids comprising a sequence selected from SEQ ID NOS: 14-25, 27, 28, 31, 32, 35, 36, 38, 40, 43-46, 48, 51-56, 58-68, 70-72, 81 and 89 is fused.\",\n \"7.A method according to claim 4 wherein at least one amino acid of the peptide sequence tag is histidine.\",\n \"8.A method according to claim 7 wherein the peptide sequence tag has the formula His-X in which X is selected from -Gly-, -His-, -Tyr-, -Gly-, -Trp-, -Val-, -Leu-, -Ser-, -Lys-, -Phe-, -Met-, -Ala-, -Glu-, -Ile-, -Thr-, -Asp-, -Asn-, -Gln-, -Arg-, -Cys- and -Pro-.\",\n \"9.A method according to claim 7 wherein the peptide sequence tag has the formula Y-His.\",\n \"10.A method according to claim 9 wherein Y is selected from -Gly-, -Ala-, -His-, and -Tyr-.\",\n \"11.A method according to claim 1 wherein the recombinant cell expresses biotin ligase and step (ii) is effected in the presence of biotin such that biotinylation occurs in vivo in said cell.\",\n \"12.A method according to claim 11 wherein recombinant cell expresses biotin.\",\n \"13.A method according to claim 1 wherein step (iii) is effected using a further antibody or a binding fragment thereof, which is specific for the peptide of SEQ ID NO:1 or a peptide of 13-50 amino acids comprising a sequence selected from SEQ ID NOS: 14-25, 27, 28, 31, 32, 35, 36, 38, 40, 43-46, 48, 51-56, 58-68, 70-72, 81 and 89.14.A method according claim 4 wherein step (iii) is effected using a further antibody or a binding fragment thereof, which is specific for the said second peptide sequence.\",\n \"15.A method according to claim 13 wherein said further antibody or binding fragment thereof is immobilised on a column, magnetic bead or loaded into a pipette tip.\",\n \"16.A method according to claim 15 wherein bound fusion protein is subsequently eluted by increasing the pH conditions.\",\n \"17.A method according to claim claim 1 wherein in step (iii) the fusion protein is isolated using a separation material which releasably binds biotin.\",\n \"18.A method according to claim 17 wherein the separation material is a modified version of avidin or streptavidin, which has lower affinity for biotin than native avidin or streptavidin.\",\n \"19.A method according to claim 17 wherein the separation material is attached to magnetic beads or pipette tips.\",\n \"20.A method according to claim 17 wherein the fusion protein is eluted from the separation material by changing the pH conditions.\",\n \"21.A method according to claim 1 wherein some areas of the coated support used in step (iv) are blocked to prevent binding of the fusion protein thereto.\",\n \"22.A method according to claim 1 wherein the peptide is a peptide of 15 amino acids in length.\",\n \"23.A method according to claim 22 wherein the peptide is of SEQ ID NO:2 Gly Leu Asn Asp lie Phe Glu Ala Gln Lys Ile Glu Trp His Glu (SEQ ID NO:2).\",\n \"24.A method according to any one of tho preceding claims claim 1 wherein the fusion protein comprises an antigen.\",\n \"25.A method according to claim 24 wherein an antigen library is used to create the array.\",\n \"26.A method according to claim 1 wherein the fusion protein comprises an antibody binding protein.\",\n \"27.A method according to claim 26 wherein the antibody binding protein is one or more of Protein A, Protein G and Protein L. 28.A method according to claim 23 wherein the antibody binding protein comprises a mixture of Protein A, Protein G and Protein L. 29.A method according to claim 26 wherein the antibody binding protein may be fused to the said peptide at the N-terminus thereof or it may be fused to said peptide at the C-terminus thereof.\",\n \"30.A method according to claim 1 wherein prior to step (iv), the identity of the expressed fusion protein is confirmed.\",\n \"31.A method according to claim 30 wherein the identity is confirmed using mass spectrometry.\",\n \"32.A method according to claim 1 wherein protein normalisation is carried out by detecting the peptide of SEQ ID NO:1 or a peptide of 13-50 amino acids comprising a sequence selected from SEQ ID NOS: 14-25, 27, 28, 31, 32, 35, 36, 38.40, 43-46, 48, 51-56, 58-68, 70-72, 81 and 89 in the fusion protein which acts as an internal control.\",\n \"33.A method according to claim 1 wherein protein normalisation is carried out by detecting the peptide sequence tag comprising between 1 and 30 amino acids in the fusion protein which acts as an internal control.\",\n \"34.A method according to claim 32 wherein the peptide is detected by an antibody with a high affinity for the said peptide.\",\n \"35.A method according to claim 32 wherein the protein normalisation is effected by performing an immunoassay simultaneously with subsequent analysis of a biological sample using the array.\",\n \"36.A method according to claim 1 wherein the avidin or streptavidin coated non-porous support used in step (iv) is a glass or plastics material.\",\n \"37.A method according to claim 1 wherein a further acceptor layer is provided on top of the foundation of the streptavidin layer on the support.\",\n \"38.A method according to claim 1 wherein the array comprises from 3-10,000 different fusion proteins.\",\n \"39.A method according to claim 38 wherein each protein is present in a form in which the peptide including SEQ ID NO:1 or a peptide of 13-50 amino acids comprising a sequence selected from SEQ ID NOS: 14-25, 27, 28, 31, 32, 35.36.38, 40, 43-46, 48, 51-56, 58-68, 70-72, 81 and 89 is fused to the C-terminus, and also in a form in which the peptide including SEQ ID NO:1 or a peptide of 13-50 amino acids comprising a sequence selected from SEQ ID NOS: 14-25, 27, 28, 31, 32, 35, 36, 38, 40, 43-46.48, 51-56, 58-68, 70-72, 81 and 89 is fused to the N-terminus.\",\n \"40.A protein array obtained by a method according to claim 1.41.A method of detecting binding between an antibody and an antigen, said method comprising the steps of: (vi) applying to the array according to claim 40 a sample which contains or is suspected of containing an antibody in the case of an array of step (v)(a), or an antigen in the case of the array of step (v) (b); and (vii) detecting bound antibody or antigen on the support.\",\n \"42.A method according to claim 41 wherein step (vii) is carried out by ELISA methods.\",\n \"43.A method according to claim 41 wherein the fusion protein array continues to be monitored for quality and/or the density of the protein during step (vi) and/or step (vii).\",\n \"44.A method according to claim 43 wherein the monitoring is effected by detecting the peptide which comprises SEQ ID NO:1 or a peptide of 13-50 amino acids comprising a sequence selected from SEQ ID NOS: 14-25, 27, 28, 31, 32, 35, 36, 38, 40, 43-46, 48, 51-56, 58-68, 70-72, 81 and 89.45.A method according to claim 43 wherein array comprises fusion proteins which further comprise a second peptide sequence, and monitoring is effected by detecting the presence of the second peptide sequence, wherein the second peptide sequence comprises between 1 and 30 amino acids.\",\n \"46.A method according to claim 1 wherein at least some of the steps are operated automatically.\",\n \"47.A method according to claim 46 wherein all the steps of the method are operated automatically.\",\n \"48.A fusion protein comprising an antibody binding protein fused at the N— or C-terminus to a peptide of 13 to 50 amino acids, which comprises SEQ ID NO:1 or a peptide of 13-50 amino acids comprising a sequence selected from SEQ ID NOS: 14-25, 27, 28, 31, 32, 35, 36, 38, 40, 43-46, 48, 51-56, 58-68, 70-72, 81 and 89.49.A fusion protein according to claim 48 wherein the peptide is a peptide of SEQ ID NO:2.50.A fusion protein according to claim 48 further comprising a second peptide sequence which acts as a tag sequence to the fusion protein wherein the sequence comprises between 1 and 20 amino acids.\",\n \"51.A fusion protein according to claim 48 wherein the antibody binding protein is Protein A, G or L or a mixture thereof.\",\n \"52.A nucleic acid sequence, which encodes the fusion protein according to claim 48.53.A nucleic acid according to claim 52 wherein the sequence which encodes the peptide is of SEQ ID NO:9 GGCCTGAACGACATCTTCGAGGCTCAGAAAATCGAATGGCACGAA (SEQ ID NO:9).\"\n ],\n [\n \"<SOH> BACKGROUND <EOH>The concept of attaching a number of different proteins to surface supports to form an “array” of proteins has been widely described in the literature (see for example EP0063810, WO84/03151, U.S. Pat.\",\n \"No.\",\n \"5,143,854).\",\n \"Recently, there has been a growing interest in the concept of manufacturing devices whereby large numbers of proteins of various classes are arrayed onto different types of solid supports.\",\n \"Examples include antigen, antibody, protein (protein-protein interaction) and functional enzymes arrays.\",\n \"The background to the technology, and the potential uses for such devices, are thoroughly catalogued in the literature (Joos et al Electrophoresis 2000, 21, 2641-2650, Haab et al Genome Biology 2000 1(6), Borrebaeck Immunology Today, August 2000) and examples of potential utility can be found in a number of recent patent applications including WO 00/07024, WO 99/40434, WO 99/39210 and WO00/54046.The concept of creating antigen arrays was described in EP 0063810 in 1982.It was reported that antigens and antibodies could be bound to a porous solid support enabling an unlimited number of antibody-antigen interactions to be conducted simultaneously.\",\n \"To make antigen arrays, antigens were simply aliquoted in very small volumes onto nitrocellulose membranes or similar supports, allowed to adsorb and then probed with the corresponding antibodies.\",\n \"As with Enzyme Linked Immunosorbent Assay (ELISA) protocols performed in solution or in plastic plates, non-specific interactions were blocked with Bovine Serum Albumin (BSA), and this is now standard practise.\",\n \"It was also reported that the elements (or spots) of the array did not diffuse and were adsorbed tightly onto the membrane.\",\n \"It should be noted, however, that the dimensions for these elements were considerably larger than those obtained in micro array device systems.\",\n \"EP 0063810 describes how the protein arrays could be made by aliquoting proteins by hand, using mechanical procedures including a “charged drop” or lithographic process.\",\n \"In this manner elements with a diameter of less than 500 microns (compared with 100 microns that can be achieved with current automated array systems) were produced.\",\n \"However, one of the main disadvantages associated with the use of membranes as opposed to non-porous surfaces is that the elements tend to diffuse through the support material unless there is immediate binding.\",\n \"Attempts were made to overcome this problem.\",\n \"U.S. Pat.\",\n \"No.\",\n \"4,496,654 describes use of porous surfaces such as paper disks which were treated with streptavidin (which is adsorbed onto the surface) enabling arrays of biotinylated antibodies to be arranged in any desired pattern.\",\n \"Following blocking with BSA, the paper discs could be probed with the antigen (exemplified with human chorionic gonadotropin) which could then be detected with an enzyme assay.\",\n \"The biotinylated antibody immediately bound very tightly to the surface of the paper reducing diffusion of the spots.\",\n \"To achieve this using an “acceptor” surface such as an avidin or streptavidin coated surface, requires that each antibody and antigen, which is attached to the array, must be biotinylated prior to attachment to the array with no guarantee that this process will not impair its avidity (or antigenicity if an antigen is used) compared with the native protein.\",\n \"Non-porous surfaces also have the disadvantage that they are not as robust as solid surfaces, including various types of glass or plastics, and so cannot be washed or treated as stringently.\",\n \"For antigen and antibody arrays, it has been found however that attaching a protein to a solid surface generally leads to a reduction in antigenicity of the antigen and avidity of the antibody compared with that observed when the antigen or antibody is in free solution.\",\n \"Previous attempts (see WO84/03 151 and Haab et al 2000 supra.)\",\n \"to immobilise antigens and antibodies were not greatly successful.\",\n \"WO84/03 151 describes that antibodies can be applied directly onto glass surfaces such as a microscope cover slip and dried.\",\n \"When blocked and then exposed to antigens, in this case in the form of whole cells, the antigens were captured by the array.\",\n \"However, WO84/03 151 further describes that these antigens needed to be added at a higher concentration compared with the equivalent ELISA performed in solution.\",\n \"It was also noted that the antibodies had to be “highly enriched in order to achieve a sufficiently dense antibody coat for the desired cell adherence”.\",\n \"It also took considerable time for the antibodies to be adsorbed onto the glass surface.\",\n \"Other approaches for the direct immobilisation of antigens and antibodies have been reported.\",\n \"One approach was to first adsorb calcium phosphate in the form of hydroxyapatite onto filter papers onto which proteins were bound by ionic interaction as described in U.S. Pat.\",\n \"No.\",\n \"5,827,669.These inventors reported that this was not effective for acidic proteins and that the antibodies suffered from “bad orientation” onto the Ca/phosphate layer.\",\n \"Success was, however, reported when this method was used in immobilising streptavidin.\",\n \"Another method for immobilising proteins to solid, non-porous surfaces included attaching them using an adhesive polyphenolic protein isolated from muscles as described in U.S. Pat.\",\n \"No.\",\n \"5,817,470.By coating solid surfaces, such as a polystyrene multi-well plate with polyphenolic protein, various antigens could be bound to the treated support and detected in an ELISA sandwich comprising of a primary antibody followed by a secondary antibody conjugated to an enzyme.\",\n \"However, the inventors conceded that the procedure was limited by the amount of antigen bound or adsorbed to the solid surface.\",\n \"The final amount of antigen strongly bound to the surface of the plate varied depending on a number of factors such as the molecular characteristics of the antigens, the properties of the solid support, the concentration of the antigen in the solution as well as the characteristics of the buffer used to dissolve the antigen used to coat or to activate the surface.\",\n \"In general, only a small fraction of the antigen present in the coating solution was adsorbed to the surface.\",\n \"Direct attachment of antibodies and antigens to non-porous surfaces was also been attempted with a collection of 113 antibodies and their corresponding antigens (Haab et al, 2000 Genome Biology 1(6)).\",\n \"By exploiting technology developed for DNA microarrays, glass slides coated with poly-s-lysine were used to immobilise both antigens in one experiment and antibodies in another.\",\n \"The results reported showed that only 50% of the arrayed antigens and 20% of the arrayed antibodies, provided specific and accurate measurements of their cognate ligands at or below concentrations of 1.60 μl/ml and 0.34 μg/ml respectively.\",\n \"The high failure rate in binding antibodies to solid surfaces would not be acceptable for a large-scale antibody array manufacturing programme.\",\n \"This supports the view that direct attachment of antigens and antibodies is an unsuitable technique to retain antibody/antigen functionality if protein arrays are to fulfil their potential.\",\n \"The use of coatings such as avidin and streptavidin as binders for biotin labelled proteins is well known for use in conjunction with many proteins.\",\n \"The proteins are generally isolated first, and then biotinylated.\",\n \"Biotin can be conjugated to the protein at any or all active lysine sites contained within it.\",\n \"Thus, when antigens or antibodies are biotinylated in this way, biotin groups may be present at their N-terminal groups and at any number of potential active lysine residues over their surface.\",\n \"This means that they will adopt any number of different orientations once bound to the streptavidin layer and so the binding properties will be diverse.\",\n \"Furthermore, access to the antigen or antibody immobilised via streptavidin will be reduced by steric hindrance, leading to generally inadequate assay.\",\n \"It has been found that it is possible to reduce the steric hindrance and increase the sensitivity of the immunoassay by including a linker between the antigen/antibody and the biotinylated site.\",\n \"U.S. Pat.\",\n \"No.\",\n \"5,811,246 describes how small synthetic peptides used in either immunoassays or for raising antisera can be linked to a “carrier” protein such as avidin or streptavidin via a linker such as various bradykinin derivatives.\",\n \"This has several advantages.\",\n \"Firstly, the condensation reaction between the free N-terminal group on the peptide and the linker preserves the charged residues essential for recognition by an antibody (immunoassay) or to elicit an immune response (immunisation).\",\n \"Secondly, the bradykinin linker can then be biotinylated in such a way as to preserve the free charged groups on the small peptide.\",\n \"In each case, the presence of the linker appears to promote a more sensitive immunoassay and an improved immune response when used as an immunising agent.\",\n \"This use of a bradykinin derivative in this way however introduces further steps and complications into the process.\",\n \"Biotinylated peptides fused to peptides or proteins of interest are described in U.S. Pat.\",\n \"Nos.\",\n \"5,723,584, 5,874,239 and 5,932,433, and further in Beckett et a!.\",\n \"Protein Sci.\",\n \"(1999) 8(4) 921-9.These peptides are used in order to biotinylate recombinant proteins, so as to allow rapid purification, immobilization, labelling and detection thereof.\",\n \"It is not suggested that these peptides should be used in particular with antigens or antibody binding proteins, or that they should be formulated in arrays.\",\n \"The present applicants have found that the peptides used in these patents allow the production of very good antigen or antibody arrays, which can be efficiently produced on non-porous supports whilst substantially retaining the binding avidity of these proteins.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>According to a first aspect of the present invention there is provided a method of forming an array of proteins selected from antigens or antibodies; said method comprising the steps of (i) expressing in a recombinant cell, a fusion protein which comprises either (a) an antigen or (b) an antibody binding protein, fused to a peptide having up to 50 amino acids, which peptide comprised amino acid sequence of SEQ ID NO 1 LX 1 X 2 IX 3 X 4 X 5 X 6 KX 7 X 8 X 9 X 10 (SEQ ID NO 1) where X1 is a naturally occurring amino acid, X 2 is any naturally occurring amino acid other than leucine, valine, isoleucine, tryptophan, phenylalanine or tyrosine, X 3 is phenylalanine or leucine, X 4 is glutamine or asparagine, X 5 is alanine, glycine, serine or threonine, X 6 is glycine or methionine, X 7 is isoleucine, methionine or valine, X 8 is glutamine, leucine, valine, tyrosine or isoleucine, X 9 is tryptophan, tyrosine, valine, phenylalanine, leucine or isoleucine and X 10 is any naturally occurring amino acid other than asparagine or glutamine; where said peptide is capable of being biotinylated by a biotin ligase at the lysine residue adjacent to X 6 ; (ii) biotinylating said peptide of the fusion protein at the lysine residue adjacent X 6 ; (iii) isolating the biotinylated fusion protein; (iv) applying the biotinylated fusion protein to an avidin or streptavidin coated non-porous support; (v) forming an array of at least three different proteins on the support by either (a) where the fusion protein comprises an antigen, carrying out steps (i) to (iv) the desired number of times to form an antigen array; or (b) where the fusion protein comprises an antibody binding protein, applying to said protein, either prior to or after step (iv) a plurality of different antibodies or binding fragments thereof.\",\n \"The applicants have found that by using a fusion of the antigen or antibody binding protein to a peptide of SEQ ID NO 1, these proteins may be immobilised onto solid surfaces, whilst substantially maintaining the antigenicity of proteins, or the binding capabilities of the antibody binding proteins.\",\n \"This may be because the fusion peptide is biotinylated rather than the protein itself, and so there is less disruption of the protein's antigenicity when attached to the support surface.\",\n \"In addition, the peptide including SEQ ID NO 1 appears to reduce steric hindrance to enable interaction between antigen and antibody.\",\n \"By ensuring that the peptide linker is attached at a terminal region of the protein, and contains the biotinylation site, sites on the protein which are essential for function appear to be largely unaffected.\",\n \"This combination is particularly advantageous in the context of methods of analysis using antigens or antibody arrays.\",\n \"The method described herein represents the first time that the mode of attachment of proteins to non-porous surfaces (step vi), the mode of protein isolation from cell lysate (step iii) and the method of biotinylation (step ii) utilise the same fusion peptide.\",\n \"Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.\",\n \"As used herein the expression “antibody binding protein” refers to proteins which are known to bind to regions of antibodies, or to mixtures of these.\",\n \"Examples of such proteins include Protein A, Protein L and Protein G Antibody binding proteins are used in accordance with the invention in the production of antibody arrays.\",\n \"The antibodies are bound by antibody binding proteins, such as Proteins A, G and/or L or a mixture of one or more of these, which are themselves anchored via the linker to the streptavidin coating on the support surface.\",\n \"While biotinylated versions of native Protein A, G and L are commercially available and can be attached to the streptavidin coating on the support surface, the applicants have found that by fusing these proteins to biotinylated tags in accordance with the present invention at the C and/or N-terminals, highly effective binding of antibodies of various types was achieved.\",\n \"This may also be the result of reduced steric factors, or that the binding sites on the proteins are all readily available.\",\n \"In addition, by using the method of the invention, the biotinylated fusion protein is immediately captured on application to the avidin or streptavidin coated support in step (iv) leading to very discrete spots of protein on the support, with minimal observable diffusion.\",\n \"Particular examples of peptides having up to 50 amino acids, which peptide comprises an amino acid sequence of SEQ ID NO 1 are listed in U.S. Pat.\",\n \"No.\",\n \"5,723,584, U.S. Pat.\",\n \"No.\",\n \"5,874,239 and U.S. Pat.\",\n \"No.\",\n \"5,932,433, the content of which are incorporated herein by reference.\",\n \"Examples of peptides provided in these references are listed below: Leu Glu Glu Val Asp Ser Thr Ser (SEQ ID NO: 14) Ser Ala Ile Phe Asp Ala Met Lys Met Val Trp Ile Ser Pro Thr Glu Phe Arg; Gln Gly Asp Arg Asp Glu Thr Leu (SEQ ID NO: 15) Pro Met Ile Leu Arg Ala Met Lys Met Glu Val Tyr Asn Pro Gly Gly His Glu Lys; Ser Lys Cys Ser Tyr Ser His Asp (SEQ ID NO: 16) Leu Lys Ile Phe Glu Ala Gln Lys Met Leu Val His Ser Tyr Leu Arg Val Met Tyr Asn Tyr; Met Ala Ser Ser Asp Asp Gly Leu (SEQ ID NO: 17) Leu Thr Ile Phe Asp Ala Thr Lys Met Met Phe Ile Arg Thr; Ser Tyr Met Asp Arg Thr Asp Val (SEQ ID NO: 18) Pro Thr Ile Leu Glu Ala Met Lys Met Glu Leu His Thr Thr Pro Trp Ala Cys Arg; Ser Phe Pro Pro Ser Leu Pro Asp (SEQ ID NO: 19) Lys Asn Ile Phe Glu Ala Met Lys Met Tyr Val Ile Thr; Ser Val Val Pro Glu Pro Gly Trp (SEQ ID NO: 20) Asp Gly Pro Phe Glu Ser Met Lys Met Val Tyr His Ser Gly Ala Gln Ser Gly Gln; Val Arg His Leu Pro Pro Pro Leu (SEQ ID NO: 21) Pro Ala Leu Phe Asp Ala Met Lys Met Glu Phe Val Thr Ser Val Gln Phe; Asp Met Thr Met Pro Thr Gly Met (SEQ ID NO: 22) Thr Lys Ile Phe Glu Ala Met Lys Met Glu Val Ser Thr; Ala Thr Ala Gly Pro Leu His Glu (SEQ ID NO: 23) Pro Asp Ile Phe Leu Ala Met Lys Met Glu Val Val Asp Val Thr Asn Lys Ala Gly Gln; Ser Met Trp Glu Thr Leu Asn Ala (SEQ ID NO: 24) Gln Lys Thr Val Leu Leu; Ser His Pro Ser Gln Leu Met Thr (SEQ ID NO: 25) Asn Asp Ile Phe Glu Gly Met Lys Met Leu Tyr His; Thr Ser Glu Leu Ser Lys Leu Asp (SEQ ID NO: 27) Ala Thr Ile Phe Ala Ala Met Lys Met Gln Trp Trp Asn Pro Gly; Val Met Glu Thr Gly Leu Asp Leu (SEQ ID NO: 28) Arg Pro Ile Leu Thr Gly Met Lys Met Asp Trp Ile Pro Lys; Leu His His Ile Leu Asp Ala Gln (SEQ ID NO: 30) Lys Met Val Trp Asn His Arg; Pro Gln Gly Ile Phe Glu Ala Gln (SEQ ID NO: 31) Lys Met Leu Trp Arg Ser; Leu Ala Gly Thr Phe Glu Ala Leu (SEQ ID NO: 32) Lys Met Ala Trp His Glu His; Leu Asn Ala Ile Phe Glu Ala Met (SEQ ID NO: 33) Lys Met Glu Tyr Ser Gly; Leu Gly Gly Ile Phe Glu Ala Met (SEQ ID NO: 34) Lys Met Glu Leu Arg Asp; Leu Leu Arg Thr Phe Glu Ala Met (SEQ ID NO: 35) Lys Met Asp Trp Arg Asn Gly; Leu Ser Thr Ile Met Glu Gly Met (SEQ ID NO: 36) Lys Met Tyr Ile Gln Arg Ser; Leu Ser Asp Ile Phe Glu Ala Met (SEQ ID NO: 37) Lys Met Val Tyr Arg Pro Cys; Leu Glu Ser Met Leu Glu Ala Met (SEQ ID NO: 38) Lys Met Gln Trp Asn Pro Gln; Leu Ser Asp Ile Phe Asp Ala Met (SEQ ID NO: 39) Lys Met Val Tyr Arg Pro Gln; Leu Ala Pro Phe Phe Glu Ser Met (SEQ ID NO: 40) Lys Met Val Trp Arg Glu His; Leu Lys Gly Ile Phe Glu Ala Met (SEQ ID NO: 41) Lys Met Glu Tyr Thr Ala Met; Leu Glu Gly Ile Phe Glu Ala Met (SEQ ID NO: 42) Lys Met Glu Tyr Ser Asn Ser; Leu Leu Gln Thr Phe Asp Ala Met (SEQ ID NO: 43) Lys Met Glu Trp Leu Pro Lys; Val Phe Asp Ile Leu Glu Ala Gln (SEQ ID NO: 44) Lys Val Val Thr Leu Arg Phe; Leu Val Ser Met Phe Asp Gly Met (SEQ ID NO: 45) Lys Met Glu Trp Lys Thr Leu; Leu Glu Pro Ile Phe Glu Ala Met (SEQ ID NO: 46) Lys Met Asp Trp Arg Leu Glu; Leu Lys Glu Ile Phe Glu Gly Met (SEQ ID NO: 47) Lys Met Glu Phe Val Lys Pro; Leu Gly Gly Ile Glu Ala Gln Lys (SEQ ID NO: 48) Met Leu Leu Tyr Arg Gly Asn; Arg Pro Val Leu Glu Asn Ile Phe (SEQ ID NO: 50) Glu Ala Met Lys Met Glu Val Trp Lys Pro; Arg Ser Pro Ile Ala Glu Ile Phe (SEQ ID NO: 51) Glu Ala Met Lys Met Glu Tyr Arg Glu Thr; Gln Asp Ser Ile Met Pro Ile Phe (SEQ ID NO: 52) Glu Ala Met Lys Met Ser Trp His Val Asn; Asp Gly Val Leu Phe Pro Ile Phe (SEQ ID NO: 53) Glu Ala Met Lys Met Ile Arg Leu Glu Thr; Val Ser Arg Thr Met Thr Asn Phe (SEQ ID NO: 54) Glu Ala Met Lys Met Ile Tyr His Asp Leu; Asp Val Leu Leu Pro Thr Val Phe (SEQ ID NO: 55) Glu Ala Met Lys Met Tyr Ile Thr Lys; Pro Asn Asp Leu Glu Arg Ile Phe (SEQ ID NO: 56) Asp Ala Met Lys Ile Val Thr Val His Ser; Thr Arg Ala Leu Leu Glu Ile Phe (SEQ ID NO: 57) Asp Ala Gln Lys Met Leu Tyr Gln His Leu; Arg Asp Val His Val Gly Ile Phe (SEQ ID NO: 58) Glu Ala Met Lys Met Tyr Thr Val Glu Thr; Gly Asp Lys Leu Thr Glu Ile Phe (SEQ ID NO: 59) Glu Ala Met Lys Ile Gln Trp Thr Ser Gly; Leu Glu Gly Leu Arg Ala Val Phe (SEQ ID NO: 60) Glu Ser Met Lys Met Glu Leu Ala Asp Glu; Val Ala Asp Ser His Asp Thr Phe (SEQ ID NO: 61) Ala Ala Met Lys Met Val Trp Leu Asp Thr; Gly Leu Pro Leu Gln Asp Ile Leu (SEQ ID NO: 62) Glu Ser Met Lys Ile Val Met Thr Ser Gly; Arg Val Pro Leu Glu Ala Ile Phe (SEQ ID NO: 63) Glu Gly Ala Lys Met Ile Trp Val Pro Asn Asn; Pro Met Ile Ser His Lys Asn Phe (SEQ ID NO: 64) Glu Ala Met Lys Met Lys Phe Val Pro Glu; Lys Leu Gly Leu Pro Ala Met Phe (SEQ ID NO: 65) Glu Ala Met Lys Met Glu Trp His Pro Ser; Gln Pro Ser Leu Leu Ser Ile Phe (SEQ ID NO: 66) Glu Ala Met Lys Met Gln Ala Ser Leu Met; Leu Leu Glu Leu Arg Ser Asn Phe (SEQ ID NO: 67) Glu Ala Met Lys Met Glu Trp Gln Ile Ser; Asp Glu Glu Leu Asn Gln Ile Phe (SEQ ID NO: 68) Glu Ala Met Lys Met Tyr Pro Leu Val His Val Thr Lys; Ser Asn Leu Val Ser Leu Leu His (SEQ ID NO: 70) Ser Gln Lys Ile Leu Trp Thr Asp Pro Gln Ser Phe Gly; Leu Phe Leu His Asp Phe Leu Asn (SEQ ID NO: 71) Ala Gln Lys Val Glu Leu Tyr Pro Val Thr Ser Ser Gly; Ser Asp Ile Asn Ala Leu Leu Ser (SEQ ID NO: 72) Thr Gln Lys Ile Tyr Trp Ala His; Met Ala Ser Ser Leu Arg Gln Ile (SEQ ID NO: 73) Leu Asp Ser Gln Lys Met Glu Trp Arg Ser Asn Ala Gly Gly Ser; Met Ala His Ser Leu Val Pro Ile (SEQ ID NO: 75) Phe Asp Ala Gln Lys Ile Glu Trp Arg Asp Pro Phe Gly Gly Ser; Met Gly Pro Asp Leu Val Asn Ile (SEQ ID NO: 76) Phe Glu Ala Gln Lys Ile Glu Trp His Pro Leu Thr Gly Gly Ser; Met Ala Phe Ser Leu Arg Ser Ile (SEQ ID NO: 77) Leu Glu Ala Gln Lys Met Glu Leu Arg Asn Thr Pro Gly Gly Ser; Met Ala Gly Gly Leu Asn Asp Ile (SEQ ID NO: 78) Phe Glu Ala Gln Lys Ile Glu Trp His Glu Asp Thr Gly Gly Ser; Met Ser Ser Tyr Leu Ala Pro Ile (SEQ ID NO: 79) Phe Glu Ala Gln Lys Ile Glu Trp His Ser Ala Tyr Gly Gly Ser; Met Ala Lys Ala Leu Gln Lys Ile (SEQ ID NO: 80) Leu Glu Ala Gln Lys Met Glu Trp Arg Ser His Pro Gly Gly Ser; Met Ala Phe Gln Leu Cys Lys Ile (SEQ ID NO: 81) Phe Tyr Ala Gln Lys Met Glu Trp His Gly Val Gly Gly Gly Ser; Met Ala Gly Ser Leu Ser Thr Ile (SEQ ID NO: 82) Phe Asp Ala Gln Lys Ile Glu Trp His Val Gly Lys Gly Gly Ser; Met Ala Gln Gln Leu Pro Asp Ile (SEQ ID NO: 83) Phe Asp Ala Gln Lys Ile Glu Trp Arg Ile Ala Gly Gly Gly Ser; Met Ala Gln Arg Leu Phe His Ile (SEQ ID NO: 84) Leu Asp Ala Gln Lys Ile Glu Trp His Gly Pro Lys Gly Gly Ser; Met Ala Gly Cys Leu Gly Pro Ile (SEQ ID NO: 85) Phe Glu Ala Gln Lys Met Glu Trp Arg His Phe Val Gly Gly Ser; Met Ala Trp Ser Leu Lys Pro Ile (SEQ ID NO: 86) Phe Asp Ala Gln Lys Ile Glu Trp His Ser Pro Gly Gly Gly Ser; Met Ala Leu Gly Leu Thr Arg Ile (SEQ ID NO: 87) Leu Asp Ala Gln Lys Ile Glu Trp His Arg Asp Ser Gly Gly Ser; Met Ala Gly Ser Leu Arg Gln Ile (SEQ ID NO: 88) Leu Asp Ala Gln Lys Ile Glu Trp Arg Arg Pro Leu Gly Gly Ser, and; Met Ala Asp Arg Leu Ala Tyr Ile (SEQ ID NO: 89) Leu Glu Ala Gln Lys Met Glu Trp His Pro His Lys Gly Gly Ser.\",\n \"These peptides, or fragments thereof which include SEQ ID NO 1 are suitable examples of peptides for use in producing fusion proteins in step (i).\",\n \"In particular, the peptides used in the method of the invention to form the fusion protein have from 13 to 20 amino acids, and preferably about 15 amino acids.\",\n \"A particularly preferred peptide for use in the fusion protein of the invention is a 15 amino acid peptide fragment of SEQ ID NO 78 shown above.\",\n \"Specifically, a preferred peptide is of amino acid sequence SEQ ID NO 2: Gly Leu Asn Asp Ile Phe Glu Ala Gln (SEQ ID NO 2) Lys Ile Glu Trp His Glu.\",\n \"This peptide is known as AviTag™ and DNA vectors encoding this sequence are available from Avidity Inc., sold under the trade names pAN4, pAN-5 and pAN-6 (which are suitable for producing fusion proteins in which the peptide of SEQ ID NO 2 is attached at the N terminus of the protein) and pAC-4, pAC-5 and pAC-6 (which are suitable for producing fusion proteins in which the peptide of SEQ ID NO 2 is attached at the C terminus of the protein).\",\n \"The sequence of these vectors are shown hereinafter as SEQ ID NO 3, SEQ ID NO 4, SEQ ID NO 5, SEQ ID NO 6, SEQ ID NO 7 and SEQ ID NO 8 in FIGS.\",\n \"7-12 respectively.\",\n \"These vectors further include the ampicillin resistance gene bla to assist in cloning.\",\n \"However the AviTag™ sequence can also be transferred into other vector systems.\",\n \"Biotinylation can be effected in various ways, either in vivo or in vitro, for example by by co-expressing biotin ligase in the expression host, by adding biotin ligase to the cell lysate or by adding the biotin ligase to the purified protein.\",\n \"In a particularly preferred embodiment, the method utilises the ability to enzymatically biotinylate a lysine residue in the fusion peptide in vivo prior to protein isolation from the cell lysate, by co-expressing biotin ligase in the expression host.\",\n \"Usually when in vitro techniques are used, the expressed protein must first be isolated from the cell lysate and then chemically biotinylated in vitro by means well known in the art.\",\n \"This results in loss of material and random biotinylation.\",\n \"Proteins with multiple biotinylated sites have an unpredictable orientation and degree of binding onto the capture surface.\",\n \"The advantage of the method of the invention is that all expressed proteins will be uniformly attached via the same residue on the same linker to the array Thus, suitably, the recombinant cell used in step (i) of the invention is engineered such that it also expresses a biotinylating enzyme and also contains biotin, such that step (ii) is effected in vivo in said cell as illustrated diagrammatically hereinafter in FIG.\",\n \"1 .\",\n \"DNA (1), which is suitably a cloned gene encoding an antigen or an antibody binding protein is sub-cloned into a vector (2) (such as pAN4, pAN-5, pAN-6 or pAC4, pAC-5 or pAC-6) which includes a sequence (3) encoding a peptide of SEQ ID NO.\",\n \"1.The subcloned gene is then expressed in an expression system such as E. coli, which has been transformed with the vector as a fusion protein (4) comprising the antigen or antibody binding protein (5) fused to a peptide (6) of SEQ ID NO.\",\n \"1.When expressed in-vivo in the presence of constitutively expressed biotin ligase, the lysine residue on the fusion peptide (6) is enzymatically biotinylated.\",\n \"If the cell does not produce biotin, then it may be added to the culture medium in order to produce the desired result.\",\n \"This reduces the number of steps involved in the process.\",\n \"A particularly suitable host cell for use in the method of the invention are the AVB100, AVB 101 and AVB99 E. coli strains available from Avidity Inc., Denver, Colo., USA.\",\n \"These strains all have the birA gene stably integrated into the chromosome so that they express biotin ligase.\",\n \"In the case of AVB 100, overexpression of of BirA protein may be achieved by induction with L-arabinose.\",\n \"The AVB101 E. coli B strain contains the pACYC184 ColE1 compatible plasmid that over-expresses biotin ligase, the elevated levels of Biotin Ligase in the cells result in complete biotinylation of fusion proteins in vivo.\",\n \"An alternative host cell is strain AVB99 (Avidity Inc) which is an E. coli strain (XL1-Blue) containing a pACYC184 plasmid with an IPTG-inducible birA gene to overexpress biotin ligase (pBirAcm).\",\n \"Fusion proteins produced in step (i) may also be isolated and biotinylated in vitro in the usual way.\",\n \"The structure of the peptide of SEQ ID NO 1 is such that biotinylation will occur reliably at lysine adjacent X 6 within SEQ ID NO 1.In a preferred embodiment of the invention, the peptide comprising the amino acid of SEQ ID NO 1 is also used as a means of isolating the fusion protein in step (iii) of the method.\",\n \"The technology for protein expression using recombinant DNA technology is well known in the art.\",\n \"However, each protein that is expressed has a different amino acid sequence and many sequences are either difficult to express in the host of choice or their sequence is hydrophobic, and therefore insoluble, or is toxic to the host.\",\n \"Even in the simplest bacterial expression systems, inclusion bodies are often formed that are difficult to disrupt while leaving the target protein in its native active state.\",\n \"It is now common practise to fuse the target protein with another protein/peptide sequence (tag) to aid the purification process subsequent to expression.\",\n \"Examples of such fusion expression systems are now widely used and have been commercialised by several suppliers.\",\n \"Such fusion peptide sequences are attached to the amino or carboxyl terminal end of a protein sequence and are recognised by specific antibodies or affinity resins.\",\n \"The expressed proteins must be solubilised from the cellular debris sometimes requiring harsh conditions including unphysiological pH values or use of chaotropic reagents and therefore the affinity purification process must be robust enough to function under such conditions.\",\n \"By using this sequence as a means of isolating or purifying the expressed fusion protein, the need for additional purification tags is eliminated.\",\n \"Thus this sequence has a dual purpose.\",\n \"In some cases it may be desirable to use a further peptide sequence tag as a means of isolating or purifying the expressed fusion protein.\",\n \"The sequence is preferably between 1 and 30 amino acids in length.\",\n \"The peptide sequence tag sequence (20) may be located at the N-terminal or C-terminal region of the antigen or antibody binding protein as shown in FIG.\",\n \"13 .\",\n \"It is, however, preferably located at the opposite end of the antigen or antibody binding protein to which SEQ ID NO 1 is fused.\",\n \"Where the additional peptide sequence tag is located on the same terminal region as SEQ ID NO 1, it is preferably fused to the free end of SEQ ID NO 1.Many peptide sequence tags are known in the art.\",\n \"Examples of suitable peptide sequence tags for the purposes of the present invention are described in U.S. Pat.\",\n \"No.\",\n \"4,569,794A, and EP0 282 042B, the contents of which are herein incorporated by reference.\",\n \"Preferably, the peptide sequence tag comprises at least one histidine amino acid.\",\n \"Even more preferably the peptide sequence tag has the formula His-X in which X is selected from the group consisting of -Gly-, -His-, -Tyr-, -Gly-, -Trp-, -Val-, -Leu-, -Ser-, -Lys-, -Phe-, -Met-, -Ala-, -Glu-, -Ile-, -Thr-, -Asp-, -Asn-, -Gln-, -Arg-, -Cys- and -Pro-.\",\n \"Alternatively the peptide sequence tag has the formula Y-His wherein Y is selected from -Gly-, -Ala-, -His- and -Tyr-.\",\n \"Particularly suitable peptide sequence tags are described in EP 0 282 042B, and a preferred example is a hexa His tag.\",\n \"In one embodiment of the method of the invention, step (iii) is effected using a further antibody or a binding fragment thereof, which is specific for the peptide of amino acid sequence including SEQ ID NO 1.The said further antibody may be raised using conventional techniques to the peptide (7) which includes an amino acid of SEQ ID NO 1.This method is illustrated diagrammatically in FIG.\",\n \"2 .\",\n \"The said further antibody is an anti-fusion antibody (8), which may be immobilised on a column, magnetic bead (9) or pipette tip, for example using a secondary antibody which is suitably an anti-species antibody (10) or other methods described in the literature, such as using an antibody binding protein such as Protein A, Protein G or Protein L, bound to the bead (9).\",\n \"This approach is highly suited to automation and to the isolation of large numbers, but small quantities, of novel fusion proteins in parallel.\",\n \"After separation from the cell lysate residue, the bound fusion protein (4) can subsequently be eluted by increasing the pH from 7.0 to 9.0.In an alternative embodiment, the fusion protein is isolated using a separation material which has some affinity for biotin but which releases the biotin fairly readily.\",\n \"Suitably the separation material is a modified version of avidin or streptavidin, which has lower affinity for biotin than native avidin or streptavidin.\",\n \"A particular example of such a material is a modified version of avidin marketed as CaptAvidin™ by by Molecular Probes (Eugene, Oreg., USA).\",\n \"In this embodiment, the fusion protein is isolated from the cellular debris, detergents and salts etc from the culture medium, by lowering the pH of the cell lysis mixture to pH 6.0 followed by affinity purification with CaptAvidin™ attached to (a) magnetic beads or (b) pipette tips using conventional methods.\",\n \"Bound fusion protein may then be eluted from the magnetic beads or mini columns by subsequently increasing the pH from 6.0 to 9.5.Prior to the step (iv), it is preferable to confirm the identity of the expressed fusion protein.\",\n \"In one particular technique, a very low volume (10 μl) of the isolated fusion protein is removed from the microtitre plate.\",\n \"The sample is digested by trypsin (using methods well known in the art).\",\n \"The resultant peptide extract is desalted and concentrated using a ZipTip™ (Millipore, Mass., USA) or equivalent, before analysis via mass spectrometry.\",\n \"Since the sequence of the fusion protein is known, identification by MALDI spectrometry to identify the peptides is usually sufficient to confirm the identity of the fusion protein.\",\n \"This technique is widely used in protein research and is summarised by T. Rabilloud (Editor) Proteome Research: 2D gel electrophoresis and identification methods.\",\n \"Furthermore, this technique can be automated and there are a number of commercially available systems from companies including Amersham Pharmacia Biotech, Bio Rad, AbiMed and Genomic Solutions (WO074852A1) that will perform this function.\",\n \"Similarly, prior to step (iv), it is preferable that the concentration of each expressed protein should be normalised where possible to eliminate variation between elements.\",\n \"Large variations in protein density cause difficulties in interpreting the data derived from such arrays (see Ekins, Clinical Chemistry (1998) 44:9 2015-2030, U.S. Pat.\",\n \"No 5,807,755 and U.S. Pat.\",\n \"No.\",\n \"5,432,099 for a detailed discussion on the quantitative aspects of protein immunoassays and protein arrays and definitions of assay sensitivity).\",\n \"Protein normalisation can be achieved by either determining the total protein concentration and or by including internal controls in the protocols.\",\n \"In a particularly preferred embodiment of the method of the invention, the fusion tag is used as an internal control and is detected by an antibody with a high affinity for the peptide of amino acid sequence which includes SEQ ID NO 1 within the fusion protein.\",\n \"Alternatively, the fusion protein can be expressed with a further peptide sequence tag and this can be used as an internal control.\",\n \"Such a tag may be the said further peptide sequence tag such as a hexa His tag as discussed above, which is expressed as part of the biotinylated fusion protein.\",\n \"The tagged version of this fusion protein may be detected through the use of an appropriate antibody such as an anti His tag antibody.\",\n \"This can be done by performing a classic immunoassay sandwich simultaneously with, or during, a subsequent analysis of a biological sample using the array.\",\n \"Using the antibody to the biotinylated fusion peptide, it is possible to determine the content of the fusion protein per μl and as a ratio of total protein present.\",\n \"The methodology may be performed using a sheep polyclonal primary antibody and secondary antibody sandwich in which the secondary antibody is conjugated with fluorescent dye (e.g.\",\n \"goat anti-mouse antibody conjugated to Alexa 488, Molecular Probes, Eugene, USA).\",\n \"The fluorescent dye used is spectrally distinct from any used with the secondary antibody for the biological sample.\",\n \"Both processes have been optimised for automation.\",\n \"The avidin or steptavidin coated non-porous support used in step (iv) of the method of the invention is suitably a glass or plastics material.\",\n \"Such supports are well suited to the production of small concentrated arrays.\",\n \"This is important, since biological samples are generally very limited in volume, and thus very valuable.\",\n \"A minimal surface area containing the targets is required for protein arrays, while still enabling the ability to achieve the required sensitivity of the assay is desirable.\",\n \"In addition, high density of either antigen or antibody in the array produces better signal to noise ratios when used in an assay.\",\n \"Furthermore, as compared to supports with porous surfaces including membranes, non-porous supports are more physically robust, are well suited to automation and have a lower background when imaged on fluorescent scanners.\",\n \"These may be coated with avidin or streptavidin using conventional methods.\",\n \"For example, the immobilisation of streptavidin to non-porous surfaces such as polystyrene multi-well plates is well known in the art.\",\n \"In its most basic form, a solution of streptavidin is left in contact with the surface for some hours.\",\n \"Un-bound protein is then removed by washing and the residual active moieties on the plastic surface blocked with BSA or an equivalent.\",\n \"Although this approach may be passive, it is effective.\",\n \"The non-covalent binding of streptavidin to polystyrene or nitrocellulose surfaces appears to be highly stable and resistant to elevated temperatures and high concentrations of chaotropic reagents, as described in WO98/37236.Avidin can be chemically attached to glass using the N-hydroxysuccinamide active ester of avidin as described by Manning, et al.\",\n \"Biochemistry 16: 1364-1370 (1977) and can be attached to nylon via carbodiimide based coupling methods as described by Jasiewicz, et al.\",\n \"Exp.\",\n \"Cell Res.\",\n \"100: 213-217 (1976).\",\n \"In another method, high molecular weight compounds such as biotin-N-hydroxy-succinimide ester, N-biotinyl-6-aminocaproyl-N-hydroxysulfosuccinimide ester, sulfosuccinimidyl-2-(biotinamido)ethyl-1,3-dithiopropionate were biotinylated and used to coat a suitable surface.\",\n \"Avidin or streptavidin was then coated in a second layer and was retained through binding the biotin linker attached to the high molecular weight compound as described in EP0620438.Attachment of streptavidin via a layer of biotin on the support surface was further developed in WO98/59243, which describes how biotin can be attached to a surface by chemical means or by light activation at 365 nm.\",\n \"The benefit that this provides is that regions of the surface can be masked.\",\n \"The elegance of these approaches is that biotin can be covalently bound to glass surfaces and will, in turn, non-covalently bind streptavidin only in those areas of the support that have been treated.\",\n \"This enables patterns of streptavidin “acceptor” protein on the support to be manufactured if required.\",\n \"In a preferred embodiment of the invention however, the entire surface of the non-porous support is coated with avidin or streptavidin, and then areas which are not required for binding are blocked, for example by addition of bovine serum albumin (BSA).\",\n \"In this way, any non-specific interaction of fusion protein with the support is reduced.\",\n \"Proteins that have been applied directly onto glass or plastic surfaces become non-covalently bound through interactions with charged groups on the solid surface the active moieties of the solid surface (typically silanol groups in glass or charged surface residues on polystyrene).\",\n \"Such non-specific adsorption of antigens or antibodies onto the surfaces of glass and plastic significantly reduces their antigenicity and antigen binding capacity respectively.\",\n \"The avidin or streptavidin layer therefore fulfils a dual role of firstly attaching the biotinylated fusion protein (non-biotinylated proteins that co-purify do not bind enabling a further purification step) and secondly, the dense layer of streptavidin shields the biotinylated fusion protein from undesirable non-specific interactions with the support surface.\",\n \"When the fusion protein is applied to the avidin or streptavidin coating on the support in step (iv), very tight but non-covalent bonding occurs.\",\n \"Preferably, the non-porous support is coated with streptavidin.\",\n \"Biotin attachment to streptavidin is multivalent, providing a binding of very high capacity when compared to that of the antigen bound directly to the support surface.\",\n \"Once the proteins have been applied to form the array, the bonding is strong enough to withstand extensive and stringent washing without appreciable loss of fusion protein.\",\n \"This is illustrated in FIG.\",\n \"3 .\",\n \"Biotinylated fusion protein (4) is attached to the surface of the array support (12) via tight, non-covalent interaction with streptavidin (14).\",\n \"In the preferred example, streptavidin (14) is covalently bound to the support material.\",\n \"Sites on the array support material to which no streptavidin molecules are bound, are blocked by BSA or other surface modifiers (13).\",\n \"Fusion proteins bind the steptavidin (14) via the biotin label (7) on the fusion peptide (6).\",\n \"Furthermore, the avidin or streptavidin layer, whether attached directly to the surface of the support or via a biotinylated linker, is highly stable, is capable of being stored dry and can be heated and or treated with aggressive reagents without apparent loss of function (unlike most antigens and antibodies).\",\n \"Further “acceptor” layers can be constructed on top of the foundation of the streptavidin layer if required.\",\n \"These may comprise other antibody binding proteins known in the art.\",\n \"The array will have the advantage of using the concentrating effect of the streptavidin, which has multivalent sites for biotin attachment.\",\n \"This enables four times the biotin interaction with both antigen arrays and antibody arrays.\",\n \"This allows higher densities of either antigen or antibody in each element of the array which in turn means that more elements can be assembled per mm 2 while achieving the same signal (see U.S. Pat.\",\n \"No.\",\n \"5,807,755 and U.S. Pat.\",\n \"No.\",\n \"5,432,099 for a discussion of the quantitative aspects) and less surface area of the solid support is utilised.\",\n \"The advantage is that less biological sample is thus required.\",\n \"Where the array is to consist of antigens arrayed on a microscope format, it suitably contains a large number of these, for example from 3-10,000 different fusion proteins.\",\n \"These may be generated or obtained from various sources, depending upon the intended nature and target for the analysis to be conducted using the array.\",\n \"Preferably, however, each protein will be expressed in the form of two fusion proteins, one with the peptide including SEQ ID NO 1 attached to the C-terminal, and one with the peptide including SEQ ID NO 1 attached to the N-terminal.\",\n \"In this way, the relative antigenicities of each version of the fusion protein to a complementary antibody can be assessed.\",\n \"The uses of antibody binding proteins such as Proteins A, G and L have been extensively reported.\",\n \"The binding of such proteins to antibodies is sufficiently tight to enable use in separation and detection techniques.\",\n \"These proteins are known to bind to the conserved regions of various classes of antibody.\",\n \"When the method of the invention is used to produce antibody arrays, antibody attachment is achieved by capture of the antibody via use of a layer of antibody binding proteins fused to a peptide which comprises SEQ ID NO 1.This layer preferably comprises of a mixture of biotinylated tagged Proteins A, G and L, and more preferably, some of which are fused at the C-terminal end, and some of which are labelled at the N-terminal end.\",\n \"By creating a mixture of antibody-binding proteins, a universal acceptor is created enabling the attachment of virtually any antibody, polyclonal, monoclonal, full-chain fragments, single chain antibodies and phage with antibody activity into which a Protein A, G or L site is present or has been engineered.\",\n \"Any antibody can be incorporated into the array without the need to pre-process or modify the antibody.\",\n \"When such antibody binding proteins are applied to an avidin or streptavidin coated surface in step (iv) a very high density of bound protein (biotinylated tagged protein binding to multivalent streptavidin) results.\",\n \"This method has the advantage that only one amino acid residue is biotinylated, and this is part of the fusion peptide, leaving the antibody binding sites on the lectins available.\",\n \"These antibody binding proteins effectively create a second layer on the solid surface.\",\n \"This layer comprising of bound fusion tagged Proteins A, G and L, acts as a universal acceptor surface for any antibody ( FIG.\",\n \"6 ) without the need for direct biotinylation of the antibody.\",\n \"This saves time, antibody and eliminates the possible degradation of the antibody's binding to its corresponding antigen.\",\n \"In a preferred embodiment, a molar excess of antibodies is pre-mixed with biotinylated peptide antibody-binding protein (e.g.\",\n \"Proteins A, G or L) fusion and incubated for up to 15 minutes.\",\n \"This antibody-antibody binding protein mixture is then applied directly to the streptavidin covered array support.\",\n \"Alternatively, the biotinylated antibody binding protein fusion may first be applied to the streptavidin covered array support.\",\n \"Individual antibodies are then applied to the surface of the coated support to form an array.\",\n \"The array produced by either method comprises very discrete spots with minimal observable diffusion, leading to a good array for assay purposes.\",\n \"The array obtained using the method of the invention is suitably used in methods for detecting binding between antigens and antibodies.\",\n \"Thus in a second aspect, the invention provides a method of detecting binding between an antibody and an antigen, said method comprising the steps of (vi) applying to the array obtained using a method of the first aspect a sample which contains or is suspected of containing an antibody in the case of an array of step (v)(a), or an antigen in the case of the array of step (v)(b); and (vii) detecting bound antibody or antigen on the support.\",\n \"Steps (vi) and (vii) of the method of the invention are suitably carried out in a conventional manner, using well known immunlogical techniques such as ELISAs, including sandwich ELISAs using labelled and in particular fluorescently labelled antibodies.\",\n \"This is illustrated in the case of an antigen array in FIG.\",\n \"4 .\",\n \"Antigens (4) bound to the array substrate (12) via streptavidin (14) are detected with a suitable primary antibody (15).\",\n \"The signal is amplified using a suitable secondary antibody (16) conjugated to a label (17).\",\n \"The label in the preferred embodiment is a fluorescent dye, such as Alexa 488, but may be any number of other types of label that are known in the art.\",\n \"Suitably the protein array continues to be monitored for quality and in particular the density of the protein during use of protein analysis devices.\",\n \"This is achieved in accordance with a preferred embodiment of the invention by using the peptide which comprises SEQ ID NO 1 or the further peptide sequence tag such as the hexa His Tag mentioned above, or any other suitable tag which performs this function as an internal standard, in a manner similar to that described above for pre-array protein normalisation.\",\n \"Once the antigens from numerous protein preparations have been arrayed onto the support surface, the array can then be used to assess antibody quality (see WO 99/39210) or can be used to determine antibody titre in serum samples (Joos et al Electrophoresis 2000, 21, 2641-2650).\",\n \"In these instances, the relative amounts of protein between the different elements in an array can be determined by adding an internal standard to the primary sample.\",\n \"The internal standard that is preferred is a sheep polyclonal antibody raised against the fusion peptide.\",\n \"This is spiked into the primary antibody solution (either antibody or serum) and is detected by an anti-sheep secondary antibody conjugated with a suitable fluorescent dye that is spectrally distinct from the labelled secondary antibody to the primary sample.\",\n \"A two-colour image is generated using a commercially available slide imager and the signal for each element is normalised to the signal resulting from the fusion protein.\",\n \"Combined with pre-array protein content normalisation, arrays of considerable consistency can be generated.\",\n \"Preferably at least some and most preferably all of the steps of the process described above are operated automatically to increase throughput and reduce labour time and costs.\",\n \"Creating antigen arrays with many novel proteins means proteins must be attached with the minimum number of steps if the process is to be viable.\",\n \"The use of a peptide which is readily and specifically biotinylated and which can act not only as a binding protein for assay purposes, but also as a purification means and an internal control for monitoring quality, provides just such a method.\",\n \"The method of the invention allows diverse collections of proteins to be attached with universal procedures, a minimum number of steps and maximum predictability of orientation.\",\n \"The method is suitable for operation on a large-scale, for example in high-throughput screening.\",\n \"In a third aspect the invention provides a protein array on a non-porous support, obtained using the method of the first aspect of the invention.\",\n \"Some elements used in the above-described methods are novel and therefore form further aspects of the invention.\",\n \"In particular, in a fourth aspect, the invention provides a fusion protein comprising an antibody binding protein fused at the N— or C-terminus to a peptide of 13 to 50 amino acids which comprises SEQ ID NO 1, such as a peptide of SEQ ID NO 2.In particular, the antibody binding protein is Protein A, G or L and preferably a mixture thereof.\",\n \"The fusion protein may additionally comprise a further peptide sequence tag such as the hexa His tag or another suitable sequence tag, which are known to those skilled in the art as discussed above.\",\n \"Such sequence tag may be located at the N or the C-terminus of the antigen or antibody binding protein.\",\n \"It is, however, preferably located at the opposite end of the antigen or antibody binding protein to which the amino acid sequence of SEQ ID NO 1 is fused.\",\n \"Where it is located at the same terminal region as SEQ ID NO 1, the sequence tag is fused to the free end of SEQ ID NO 1.A fifth aspect of the invention comprises a nucleic acid sequence which encodes the fusion protein of the fourth aspects.\",\n \"In particular in this case, the sequence which encodes the peptide is suitably of SEQ ID NO 9.\",\n \"(SEQ ID NO 9) GGCCTGAACGACATCTTCGAGGCTCAGAAAATCGAATGGCACGAA\"\n ],\n [\n \"FIELD OF THE INVENTION The present invention relates to a method of producing arrays for conducting protein analysis, in particular of antibodies, antigens or antibody binding proteins, to protein arrays produced, methods of conducting analysis using them and novel entities incorporated in them.\",\n \"More specifically, the process relates to a method of producing a range of antibodies and/or antigens and immobilising these in an array, for use in protein or binding analysis.\",\n \"BACKGROUND The concept of attaching a number of different proteins to surface supports to form an “array” of proteins has been widely described in the literature (see for example EP0063810, WO84/03151, U.S. Pat.\",\n \"No.\",\n \"5,143,854).\",\n \"Recently, there has been a growing interest in the concept of manufacturing devices whereby large numbers of proteins of various classes are arrayed onto different types of solid supports.\",\n \"Examples include antigen, antibody, protein (protein-protein interaction) and functional enzymes arrays.\",\n \"The background to the technology, and the potential uses for such devices, are thoroughly catalogued in the literature (Joos et al Electrophoresis 2000, 21, 2641-2650, Haab et al Genome Biology 2000 1(6), Borrebaeck Immunology Today, August 2000) and examples of potential utility can be found in a number of recent patent applications including WO 00/07024, WO 99/40434, WO 99/39210 and WO00/54046.The concept of creating antigen arrays was described in EP 0063810 in 1982.It was reported that antigens and antibodies could be bound to a porous solid support enabling an unlimited number of antibody-antigen interactions to be conducted simultaneously.\",\n \"To make antigen arrays, antigens were simply aliquoted in very small volumes onto nitrocellulose membranes or similar supports, allowed to adsorb and then probed with the corresponding antibodies.\",\n \"As with Enzyme Linked Immunosorbent Assay (ELISA) protocols performed in solution or in plastic plates, non-specific interactions were blocked with Bovine Serum Albumin (BSA), and this is now standard practise.\",\n \"It was also reported that the elements (or spots) of the array did not diffuse and were adsorbed tightly onto the membrane.\",\n \"It should be noted, however, that the dimensions for these elements were considerably larger than those obtained in micro array device systems.\",\n \"EP 0063810 describes how the protein arrays could be made by aliquoting proteins by hand, using mechanical procedures including a “charged drop” or lithographic process.\",\n \"In this manner elements with a diameter of less than 500 microns (compared with 100 microns that can be achieved with current automated array systems) were produced.\",\n \"However, one of the main disadvantages associated with the use of membranes as opposed to non-porous surfaces is that the elements tend to diffuse through the support material unless there is immediate binding.\",\n \"Attempts were made to overcome this problem.\",\n \"U.S. Pat.\",\n \"No.\",\n \"4,496,654 describes use of porous surfaces such as paper disks which were treated with streptavidin (which is adsorbed onto the surface) enabling arrays of biotinylated antibodies to be arranged in any desired pattern.\",\n \"Following blocking with BSA, the paper discs could be probed with the antigen (exemplified with human chorionic gonadotropin) which could then be detected with an enzyme assay.\",\n \"The biotinylated antibody immediately bound very tightly to the surface of the paper reducing diffusion of the spots.\",\n \"To achieve this using an “acceptor” surface such as an avidin or streptavidin coated surface, requires that each antibody and antigen, which is attached to the array, must be biotinylated prior to attachment to the array with no guarantee that this process will not impair its avidity (or antigenicity if an antigen is used) compared with the native protein.\",\n \"Non-porous surfaces also have the disadvantage that they are not as robust as solid surfaces, including various types of glass or plastics, and so cannot be washed or treated as stringently.\",\n \"For antigen and antibody arrays, it has been found however that attaching a protein to a solid surface generally leads to a reduction in antigenicity of the antigen and avidity of the antibody compared with that observed when the antigen or antibody is in free solution.\",\n \"Previous attempts (see WO84/03 151 and Haab et al 2000 supra.)\",\n \"to immobilise antigens and antibodies were not greatly successful.\",\n \"WO84/03 151 describes that antibodies can be applied directly onto glass surfaces such as a microscope cover slip and dried.\",\n \"When blocked and then exposed to antigens, in this case in the form of whole cells, the antigens were captured by the array.\",\n \"However, WO84/03 151 further describes that these antigens needed to be added at a higher concentration compared with the equivalent ELISA performed in solution.\",\n \"It was also noted that the antibodies had to be “highly enriched in order to achieve a sufficiently dense antibody coat for the desired cell adherence”.\",\n \"It also took considerable time for the antibodies to be adsorbed onto the glass surface.\",\n \"Other approaches for the direct immobilisation of antigens and antibodies have been reported.\",\n \"One approach was to first adsorb calcium phosphate in the form of hydroxyapatite onto filter papers onto which proteins were bound by ionic interaction as described in U.S. Pat.\",\n \"No.\",\n \"5,827,669.These inventors reported that this was not effective for acidic proteins and that the antibodies suffered from “bad orientation” onto the Ca/phosphate layer.\",\n \"Success was, however, reported when this method was used in immobilising streptavidin.\",\n \"Another method for immobilising proteins to solid, non-porous surfaces included attaching them using an adhesive polyphenolic protein isolated from muscles as described in U.S. Pat.\",\n \"No.\",\n \"5,817,470.By coating solid surfaces, such as a polystyrene multi-well plate with polyphenolic protein, various antigens could be bound to the treated support and detected in an ELISA sandwich comprising of a primary antibody followed by a secondary antibody conjugated to an enzyme.\",\n \"However, the inventors conceded that the procedure was limited by the amount of antigen bound or adsorbed to the solid surface.\",\n \"The final amount of antigen strongly bound to the surface of the plate varied depending on a number of factors such as the molecular characteristics of the antigens, the properties of the solid support, the concentration of the antigen in the solution as well as the characteristics of the buffer used to dissolve the antigen used to coat or to activate the surface.\",\n \"In general, only a small fraction of the antigen present in the coating solution was adsorbed to the surface.\",\n \"Direct attachment of antibodies and antigens to non-porous surfaces was also been attempted with a collection of 113 antibodies and their corresponding antigens (Haab et al, 2000 Genome Biology 1(6)).\",\n \"By exploiting technology developed for DNA microarrays, glass slides coated with poly-s-lysine were used to immobilise both antigens in one experiment and antibodies in another.\",\n \"The results reported showed that only 50% of the arrayed antigens and 20% of the arrayed antibodies, provided specific and accurate measurements of their cognate ligands at or below concentrations of 1.60 μl/ml and 0.34 μg/ml respectively.\",\n \"The high failure rate in binding antibodies to solid surfaces would not be acceptable for a large-scale antibody array manufacturing programme.\",\n \"This supports the view that direct attachment of antigens and antibodies is an unsuitable technique to retain antibody/antigen functionality if protein arrays are to fulfil their potential.\",\n \"The use of coatings such as avidin and streptavidin as binders for biotin labelled proteins is well known for use in conjunction with many proteins.\",\n \"The proteins are generally isolated first, and then biotinylated.\",\n \"Biotin can be conjugated to the protein at any or all active lysine sites contained within it.\",\n \"Thus, when antigens or antibodies are biotinylated in this way, biotin groups may be present at their N-terminal groups and at any number of potential active lysine residues over their surface.\",\n \"This means that they will adopt any number of different orientations once bound to the streptavidin layer and so the binding properties will be diverse.\",\n \"Furthermore, access to the antigen or antibody immobilised via streptavidin will be reduced by steric hindrance, leading to generally inadequate assay.\",\n \"It has been found that it is possible to reduce the steric hindrance and increase the sensitivity of the immunoassay by including a linker between the antigen/antibody and the biotinylated site.\",\n \"U.S. Pat.\",\n \"No.\",\n \"5,811,246 describes how small synthetic peptides used in either immunoassays or for raising antisera can be linked to a “carrier” protein such as avidin or streptavidin via a linker such as various bradykinin derivatives.\",\n \"This has several advantages.\",\n \"Firstly, the condensation reaction between the free N-terminal group on the peptide and the linker preserves the charged residues essential for recognition by an antibody (immunoassay) or to elicit an immune response (immunisation).\",\n \"Secondly, the bradykinin linker can then be biotinylated in such a way as to preserve the free charged groups on the small peptide.\",\n \"In each case, the presence of the linker appears to promote a more sensitive immunoassay and an improved immune response when used as an immunising agent.\",\n \"This use of a bradykinin derivative in this way however introduces further steps and complications into the process.\",\n \"Biotinylated peptides fused to peptides or proteins of interest are described in U.S. Pat.\",\n \"Nos.\",\n \"5,723,584, 5,874,239 and 5,932,433, and further in Beckett et a!.\",\n \"Protein Sci.\",\n \"(1999) 8(4) 921-9.These peptides are used in order to biotinylate recombinant proteins, so as to allow rapid purification, immobilization, labelling and detection thereof.\",\n \"It is not suggested that these peptides should be used in particular with antigens or antibody binding proteins, or that they should be formulated in arrays.\",\n \"The present applicants have found that the peptides used in these patents allow the production of very good antigen or antibody arrays, which can be efficiently produced on non-porous supports whilst substantially retaining the binding avidity of these proteins.\",\n \"SUMMARY OF THE INVENTION According to a first aspect of the present invention there is provided a method of forming an array of proteins selected from antigens or antibodies; said method comprising the steps of (i) expressing in a recombinant cell, a fusion protein which comprises either (a) an antigen or (b) an antibody binding protein, fused to a peptide having up to 50 amino acids, which peptide comprised amino acid sequence of SEQ ID NO 1 LX1X2IX3X4X5X6KX7X8X9X10 (SEQ ID NO 1) where X1 is a naturally occurring amino acid, X2 is any naturally occurring amino acid other than leucine, valine, isoleucine, tryptophan, phenylalanine or tyrosine, X3 is phenylalanine or leucine, X4 is glutamine or asparagine, X5 is alanine, glycine, serine or threonine, X6 is glycine or methionine, X7 is isoleucine, methionine or valine, X8 is glutamine, leucine, valine, tyrosine or isoleucine, X9 is tryptophan, tyrosine, valine, phenylalanine, leucine or isoleucine and X10 is any naturally occurring amino acid other than asparagine or glutamine; where said peptide is capable of being biotinylated by a biotin ligase at the lysine residue adjacent to X6; (ii) biotinylating said peptide of the fusion protein at the lysine residue adjacent X6; (iii) isolating the biotinylated fusion protein; (iv) applying the biotinylated fusion protein to an avidin or streptavidin coated non-porous support; (v) forming an array of at least three different proteins on the support by either (a) where the fusion protein comprises an antigen, carrying out steps (i) to (iv) the desired number of times to form an antigen array; or (b) where the fusion protein comprises an antibody binding protein, applying to said protein, either prior to or after step (iv) a plurality of different antibodies or binding fragments thereof.\",\n \"The applicants have found that by using a fusion of the antigen or antibody binding protein to a peptide of SEQ ID NO 1, these proteins may be immobilised onto solid surfaces, whilst substantially maintaining the antigenicity of proteins, or the binding capabilities of the antibody binding proteins.\",\n \"This may be because the fusion peptide is biotinylated rather than the protein itself, and so there is less disruption of the protein's antigenicity when attached to the support surface.\",\n \"In addition, the peptide including SEQ ID NO 1 appears to reduce steric hindrance to enable interaction between antigen and antibody.\",\n \"By ensuring that the peptide linker is attached at a terminal region of the protein, and contains the biotinylation site, sites on the protein which are essential for function appear to be largely unaffected.\",\n \"This combination is particularly advantageous in the context of methods of analysis using antigens or antibody arrays.\",\n \"The method described herein represents the first time that the mode of attachment of proteins to non-porous surfaces (step vi), the mode of protein isolation from cell lysate (step iii) and the method of biotinylation (step ii) utilise the same fusion peptide.\",\n \"Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.\",\n \"As used herein the expression “antibody binding protein” refers to proteins which are known to bind to regions of antibodies, or to mixtures of these.\",\n \"Examples of such proteins include Protein A, Protein L and Protein G Antibody binding proteins are used in accordance with the invention in the production of antibody arrays.\",\n \"The antibodies are bound by antibody binding proteins, such as Proteins A, G and/or L or a mixture of one or more of these, which are themselves anchored via the linker to the streptavidin coating on the support surface.\",\n \"While biotinylated versions of native Protein A, G and L are commercially available and can be attached to the streptavidin coating on the support surface, the applicants have found that by fusing these proteins to biotinylated tags in accordance with the present invention at the C and/or N-terminals, highly effective binding of antibodies of various types was achieved.\",\n \"This may also be the result of reduced steric factors, or that the binding sites on the proteins are all readily available.\",\n \"In addition, by using the method of the invention, the biotinylated fusion protein is immediately captured on application to the avidin or streptavidin coated support in step (iv) leading to very discrete spots of protein on the support, with minimal observable diffusion.\",\n \"Particular examples of peptides having up to 50 amino acids, which peptide comprises an amino acid sequence of SEQ ID NO 1 are listed in U.S. Pat.\",\n \"No.\",\n \"5,723,584, U.S. Pat.\",\n \"No.\",\n \"5,874,239 and U.S. Pat.\",\n \"No.\",\n \"5,932,433, the content of which are incorporated herein by reference.\",\n \"Examples of peptides provided in these references are listed below: Leu Glu Glu Val Asp Ser Thr Ser (SEQ ID NO: 14) Ser Ala Ile Phe Asp Ala Met Lys Met Val Trp Ile Ser Pro Thr Glu Phe Arg; Gln Gly Asp Arg Asp Glu Thr Leu (SEQ ID NO: 15) Pro Met Ile Leu Arg Ala Met Lys Met Glu Val Tyr Asn Pro Gly Gly His Glu Lys; Ser Lys Cys Ser Tyr Ser His Asp (SEQ ID NO: 16) Leu Lys Ile Phe Glu Ala Gln Lys Met Leu Val His Ser Tyr Leu Arg Val Met Tyr Asn Tyr; Met Ala Ser Ser Asp Asp Gly Leu (SEQ ID NO: 17) Leu Thr Ile Phe Asp Ala Thr Lys Met Met Phe Ile Arg Thr; Ser Tyr Met Asp Arg Thr Asp Val (SEQ ID NO: 18) Pro Thr Ile Leu Glu Ala Met Lys Met Glu Leu His Thr Thr Pro Trp Ala Cys Arg; Ser Phe Pro Pro Ser Leu Pro Asp (SEQ ID NO: 19) Lys Asn Ile Phe Glu Ala Met Lys Met Tyr Val Ile Thr; Ser Val Val Pro Glu Pro Gly Trp (SEQ ID NO: 20) Asp Gly Pro Phe Glu Ser Met Lys Met Val Tyr His Ser Gly Ala Gln Ser Gly Gln; Val Arg His Leu Pro Pro Pro Leu (SEQ ID NO: 21) Pro Ala Leu Phe Asp Ala Met Lys Met Glu Phe Val Thr Ser Val Gln Phe; Asp Met Thr Met Pro Thr Gly Met (SEQ ID NO: 22) Thr Lys Ile Phe Glu Ala Met Lys Met Glu Val Ser Thr; Ala Thr Ala Gly Pro Leu His Glu (SEQ ID NO: 23) Pro Asp Ile Phe Leu Ala Met Lys Met Glu Val Val Asp Val Thr Asn Lys Ala Gly Gln; Ser Met Trp Glu Thr Leu Asn Ala (SEQ ID NO: 24) Gln Lys Thr Val Leu Leu; Ser His Pro Ser Gln Leu Met Thr (SEQ ID NO: 25) Asn Asp Ile Phe Glu Gly Met Lys Met Leu Tyr His; Thr Ser Glu Leu Ser Lys Leu Asp (SEQ ID NO: 27) Ala Thr Ile Phe Ala Ala Met Lys Met Gln Trp Trp Asn Pro Gly; Val Met Glu Thr Gly Leu Asp Leu (SEQ ID NO: 28) Arg Pro Ile Leu Thr Gly Met Lys Met Asp Trp Ile Pro Lys; Leu His His Ile Leu Asp Ala Gln (SEQ ID NO: 30) Lys Met Val Trp Asn His Arg; Pro Gln Gly Ile Phe Glu Ala Gln (SEQ ID NO: 31) Lys Met Leu Trp Arg Ser; Leu Ala Gly Thr Phe Glu Ala Leu (SEQ ID NO: 32) Lys Met Ala Trp His Glu His; Leu Asn Ala Ile Phe Glu Ala Met (SEQ ID NO: 33) Lys Met Glu Tyr Ser Gly; Leu Gly Gly Ile Phe Glu Ala Met (SEQ ID NO: 34) Lys Met Glu Leu Arg Asp; Leu Leu Arg Thr Phe Glu Ala Met (SEQ ID NO: 35) Lys Met Asp Trp Arg Asn Gly; Leu Ser Thr Ile Met Glu Gly Met (SEQ ID NO: 36) Lys Met Tyr Ile Gln Arg Ser; Leu Ser Asp Ile Phe Glu Ala Met (SEQ ID NO: 37) Lys Met Val Tyr Arg Pro Cys; Leu Glu Ser Met Leu Glu Ala Met (SEQ ID NO: 38) Lys Met Gln Trp Asn Pro Gln; Leu Ser Asp Ile Phe Asp Ala Met (SEQ ID NO: 39) Lys Met Val Tyr Arg Pro Gln; Leu Ala Pro Phe Phe Glu Ser Met (SEQ ID NO: 40) Lys Met Val Trp Arg Glu His; Leu Lys Gly Ile Phe Glu Ala Met (SEQ ID NO: 41) Lys Met Glu Tyr Thr Ala Met; Leu Glu Gly Ile Phe Glu Ala Met (SEQ ID NO: 42) Lys Met Glu Tyr Ser Asn Ser; Leu Leu Gln Thr Phe Asp Ala Met (SEQ ID NO: 43) Lys Met Glu Trp Leu Pro Lys; Val Phe Asp Ile Leu Glu Ala Gln (SEQ ID NO: 44) Lys Val Val Thr Leu Arg Phe; Leu Val Ser Met Phe Asp Gly Met (SEQ ID NO: 45) Lys Met Glu Trp Lys Thr Leu; Leu Glu Pro Ile Phe Glu Ala Met (SEQ ID NO: 46) Lys Met Asp Trp Arg Leu Glu; Leu Lys Glu Ile Phe Glu Gly Met (SEQ ID NO: 47) Lys Met Glu Phe Val Lys Pro; Leu Gly Gly Ile Glu Ala Gln Lys (SEQ ID NO: 48) Met Leu Leu Tyr Arg Gly Asn; Arg Pro Val Leu Glu Asn Ile Phe (SEQ ID NO: 50) Glu Ala Met Lys Met Glu Val Trp Lys Pro; Arg Ser Pro Ile Ala Glu Ile Phe (SEQ ID NO: 51) Glu Ala Met Lys Met Glu Tyr Arg Glu Thr; Gln Asp Ser Ile Met Pro Ile Phe (SEQ ID NO: 52) Glu Ala Met Lys Met Ser Trp His Val Asn; Asp Gly Val Leu Phe Pro Ile Phe (SEQ ID NO: 53) Glu Ala Met Lys Met Ile Arg Leu Glu Thr; Val Ser Arg Thr Met Thr Asn Phe (SEQ ID NO: 54) Glu Ala Met Lys Met Ile Tyr His Asp Leu; Asp Val Leu Leu Pro Thr Val Phe (SEQ ID NO: 55) Glu Ala Met Lys Met Tyr Ile Thr Lys; Pro Asn Asp Leu Glu Arg Ile Phe (SEQ ID NO: 56) Asp Ala Met Lys Ile Val Thr Val His Ser; Thr Arg Ala Leu Leu Glu Ile Phe (SEQ ID NO: 57) Asp Ala Gln Lys Met Leu Tyr Gln His Leu; Arg Asp Val His Val Gly Ile Phe (SEQ ID NO: 58) Glu Ala Met Lys Met Tyr Thr Val Glu Thr; Gly Asp Lys Leu Thr Glu Ile Phe (SEQ ID NO: 59) Glu Ala Met Lys Ile Gln Trp Thr Ser Gly; Leu Glu Gly Leu Arg Ala Val Phe (SEQ ID NO: 60) Glu Ser Met Lys Met Glu Leu Ala Asp Glu; Val Ala Asp Ser His Asp Thr Phe (SEQ ID NO: 61) Ala Ala Met Lys Met Val Trp Leu Asp Thr; Gly Leu Pro Leu Gln Asp Ile Leu (SEQ ID NO: 62) Glu Ser Met Lys Ile Val Met Thr Ser Gly; Arg Val Pro Leu Glu Ala Ile Phe (SEQ ID NO: 63) Glu Gly Ala Lys Met Ile Trp Val Pro Asn Asn; Pro Met Ile Ser His Lys Asn Phe (SEQ ID NO: 64) Glu Ala Met Lys Met Lys Phe Val Pro Glu; Lys Leu Gly Leu Pro Ala Met Phe (SEQ ID NO: 65) Glu Ala Met Lys Met Glu Trp His Pro Ser; Gln Pro Ser Leu Leu Ser Ile Phe (SEQ ID NO: 66) Glu Ala Met Lys Met Gln Ala Ser Leu Met; Leu Leu Glu Leu Arg Ser Asn Phe (SEQ ID NO: 67) Glu Ala Met Lys Met Glu Trp Gln Ile Ser; Asp Glu Glu Leu Asn Gln Ile Phe (SEQ ID NO: 68) Glu Ala Met Lys Met Tyr Pro Leu Val His Val Thr Lys; Ser Asn Leu Val Ser Leu Leu His (SEQ ID NO: 70) Ser Gln Lys Ile Leu Trp Thr Asp Pro Gln Ser Phe Gly; Leu Phe Leu His Asp Phe Leu Asn (SEQ ID NO: 71) Ala Gln Lys Val Glu Leu Tyr Pro Val Thr Ser Ser Gly; Ser Asp Ile Asn Ala Leu Leu Ser (SEQ ID NO: 72) Thr Gln Lys Ile Tyr Trp Ala His; Met Ala Ser Ser Leu Arg Gln Ile (SEQ ID NO: 73) Leu Asp Ser Gln Lys Met Glu Trp Arg Ser Asn Ala Gly Gly Ser; Met Ala His Ser Leu Val Pro Ile (SEQ ID NO: 75) Phe Asp Ala Gln Lys Ile Glu Trp Arg Asp Pro Phe Gly Gly Ser; Met Gly Pro Asp Leu Val Asn Ile (SEQ ID NO: 76) Phe Glu Ala Gln Lys Ile Glu Trp His Pro Leu Thr Gly Gly Ser; Met Ala Phe Ser Leu Arg Ser Ile (SEQ ID NO: 77) Leu Glu Ala Gln Lys Met Glu Leu Arg Asn Thr Pro Gly Gly Ser; Met Ala Gly Gly Leu Asn Asp Ile (SEQ ID NO: 78) Phe Glu Ala Gln Lys Ile Glu Trp His Glu Asp Thr Gly Gly Ser; Met Ser Ser Tyr Leu Ala Pro Ile (SEQ ID NO: 79) Phe Glu Ala Gln Lys Ile Glu Trp His Ser Ala Tyr Gly Gly Ser; Met Ala Lys Ala Leu Gln Lys Ile (SEQ ID NO: 80) Leu Glu Ala Gln Lys Met Glu Trp Arg Ser His Pro Gly Gly Ser; Met Ala Phe Gln Leu Cys Lys Ile (SEQ ID NO: 81) Phe Tyr Ala Gln Lys Met Glu Trp His Gly Val Gly Gly Gly Ser; Met Ala Gly Ser Leu Ser Thr Ile (SEQ ID NO: 82) Phe Asp Ala Gln Lys Ile Glu Trp His Val Gly Lys Gly Gly Ser; Met Ala Gln Gln Leu Pro Asp Ile (SEQ ID NO: 83) Phe Asp Ala Gln Lys Ile Glu Trp Arg Ile Ala Gly Gly Gly Ser; Met Ala Gln Arg Leu Phe His Ile (SEQ ID NO: 84) Leu Asp Ala Gln Lys Ile Glu Trp His Gly Pro Lys Gly Gly Ser; Met Ala Gly Cys Leu Gly Pro Ile (SEQ ID NO: 85) Phe Glu Ala Gln Lys Met Glu Trp Arg His Phe Val Gly Gly Ser; Met Ala Trp Ser Leu Lys Pro Ile (SEQ ID NO: 86) Phe Asp Ala Gln Lys Ile Glu Trp His Ser Pro Gly Gly Gly Ser; Met Ala Leu Gly Leu Thr Arg Ile (SEQ ID NO: 87) Leu Asp Ala Gln Lys Ile Glu Trp His Arg Asp Ser Gly Gly Ser; Met Ala Gly Ser Leu Arg Gln Ile (SEQ ID NO: 88) Leu Asp Ala Gln Lys Ile Glu Trp Arg Arg Pro Leu Gly Gly Ser, and; Met Ala Asp Arg Leu Ala Tyr Ile (SEQ ID NO: 89) Leu Glu Ala Gln Lys Met Glu Trp His Pro His Lys Gly Gly Ser.\",\n \"These peptides, or fragments thereof which include SEQ ID NO 1 are suitable examples of peptides for use in producing fusion proteins in step (i).\",\n \"In particular, the peptides used in the method of the invention to form the fusion protein have from 13 to 20 amino acids, and preferably about 15 amino acids.\",\n \"A particularly preferred peptide for use in the fusion protein of the invention is a 15 amino acid peptide fragment of SEQ ID NO 78 shown above.\",\n \"Specifically, a preferred peptide is of amino acid sequence SEQ ID NO 2: Gly Leu Asn Asp Ile Phe Glu Ala Gln (SEQ ID NO 2) Lys Ile Glu Trp His Glu.\",\n \"This peptide is known as AviTag™ and DNA vectors encoding this sequence are available from Avidity Inc., sold under the trade names pAN4, pAN-5 and pAN-6 (which are suitable for producing fusion proteins in which the peptide of SEQ ID NO 2 is attached at the N terminus of the protein) and pAC-4, pAC-5 and pAC-6 (which are suitable for producing fusion proteins in which the peptide of SEQ ID NO 2 is attached at the C terminus of the protein).\",\n \"The sequence of these vectors are shown hereinafter as SEQ ID NO 3, SEQ ID NO 4, SEQ ID NO 5, SEQ ID NO 6, SEQ ID NO 7 and SEQ ID NO 8 in FIGS.\",\n \"7-12 respectively.\",\n \"These vectors further include the ampicillin resistance gene bla to assist in cloning.\",\n \"However the AviTag™ sequence can also be transferred into other vector systems.\",\n \"Biotinylation can be effected in various ways, either in vivo or in vitro, for example by by co-expressing biotin ligase in the expression host, by adding biotin ligase to the cell lysate or by adding the biotin ligase to the purified protein.\",\n \"In a particularly preferred embodiment, the method utilises the ability to enzymatically biotinylate a lysine residue in the fusion peptide in vivo prior to protein isolation from the cell lysate, by co-expressing biotin ligase in the expression host.\",\n \"Usually when in vitro techniques are used, the expressed protein must first be isolated from the cell lysate and then chemically biotinylated in vitro by means well known in the art.\",\n \"This results in loss of material and random biotinylation.\",\n \"Proteins with multiple biotinylated sites have an unpredictable orientation and degree of binding onto the capture surface.\",\n \"The advantage of the method of the invention is that all expressed proteins will be uniformly attached via the same residue on the same linker to the array Thus, suitably, the recombinant cell used in step (i) of the invention is engineered such that it also expresses a biotinylating enzyme and also contains biotin, such that step (ii) is effected in vivo in said cell as illustrated diagrammatically hereinafter in FIG.\",\n \"1.DNA (1), which is suitably a cloned gene encoding an antigen or an antibody binding protein is sub-cloned into a vector (2) (such as pAN4, pAN-5, pAN-6 or pAC4, pAC-5 or pAC-6) which includes a sequence (3) encoding a peptide of SEQ ID NO.\",\n \"1.The subcloned gene is then expressed in an expression system such as E. coli, which has been transformed with the vector as a fusion protein (4) comprising the antigen or antibody binding protein (5) fused to a peptide (6) of SEQ ID NO.\",\n \"1.When expressed in-vivo in the presence of constitutively expressed biotin ligase, the lysine residue on the fusion peptide (6) is enzymatically biotinylated.\",\n \"If the cell does not produce biotin, then it may be added to the culture medium in order to produce the desired result.\",\n \"This reduces the number of steps involved in the process.\",\n \"A particularly suitable host cell for use in the method of the invention are the AVB100, AVB 101 and AVB99 E. coli strains available from Avidity Inc., Denver, Colo., USA.\",\n \"These strains all have the birA gene stably integrated into the chromosome so that they express biotin ligase.\",\n \"In the case of AVB 100, overexpression of of BirA protein may be achieved by induction with L-arabinose.\",\n \"The AVB101 E. coli B strain contains the pACYC184 ColE1 compatible plasmid that over-expresses biotin ligase, the elevated levels of Biotin Ligase in the cells result in complete biotinylation of fusion proteins in vivo.\",\n \"An alternative host cell is strain AVB99 (Avidity Inc) which is an E. coli strain (XL1-Blue) containing a pACYC184 plasmid with an IPTG-inducible birA gene to overexpress biotin ligase (pBirAcm).\",\n \"Fusion proteins produced in step (i) may also be isolated and biotinylated in vitro in the usual way.\",\n \"The structure of the peptide of SEQ ID NO 1 is such that biotinylation will occur reliably at lysine adjacent X6 within SEQ ID NO 1.In a preferred embodiment of the invention, the peptide comprising the amino acid of SEQ ID NO 1 is also used as a means of isolating the fusion protein in step (iii) of the method.\",\n \"The technology for protein expression using recombinant DNA technology is well known in the art.\",\n \"However, each protein that is expressed has a different amino acid sequence and many sequences are either difficult to express in the host of choice or their sequence is hydrophobic, and therefore insoluble, or is toxic to the host.\",\n \"Even in the simplest bacterial expression systems, inclusion bodies are often formed that are difficult to disrupt while leaving the target protein in its native active state.\",\n \"It is now common practise to fuse the target protein with another protein/peptide sequence (tag) to aid the purification process subsequent to expression.\",\n \"Examples of such fusion expression systems are now widely used and have been commercialised by several suppliers.\",\n \"Such fusion peptide sequences are attached to the amino or carboxyl terminal end of a protein sequence and are recognised by specific antibodies or affinity resins.\",\n \"The expressed proteins must be solubilised from the cellular debris sometimes requiring harsh conditions including unphysiological pH values or use of chaotropic reagents and therefore the affinity purification process must be robust enough to function under such conditions.\",\n \"By using this sequence as a means of isolating or purifying the expressed fusion protein, the need for additional purification tags is eliminated.\",\n \"Thus this sequence has a dual purpose.\",\n \"In some cases it may be desirable to use a further peptide sequence tag as a means of isolating or purifying the expressed fusion protein.\",\n \"The sequence is preferably between 1 and 30 amino acids in length.\",\n \"The peptide sequence tag sequence (20) may be located at the N-terminal or C-terminal region of the antigen or antibody binding protein as shown in FIG.\",\n \"13.It is, however, preferably located at the opposite end of the antigen or antibody binding protein to which SEQ ID NO 1 is fused.\",\n \"Where the additional peptide sequence tag is located on the same terminal region as SEQ ID NO 1, it is preferably fused to the free end of SEQ ID NO 1.Many peptide sequence tags are known in the art.\",\n \"Examples of suitable peptide sequence tags for the purposes of the present invention are described in U.S. Pat.\",\n \"No.\",\n \"4,569,794A, and EP0 282 042B, the contents of which are herein incorporated by reference.\",\n \"Preferably, the peptide sequence tag comprises at least one histidine amino acid.\",\n \"Even more preferably the peptide sequence tag has the formula His-X in which X is selected from the group consisting of -Gly-, -His-, -Tyr-, -Gly-, -Trp-, -Val-, -Leu-, -Ser-, -Lys-, -Phe-, -Met-, -Ala-, -Glu-, -Ile-, -Thr-, -Asp-, -Asn-, -Gln-, -Arg-, -Cys- and -Pro-.\",\n \"Alternatively the peptide sequence tag has the formula Y-His wherein Y is selected from -Gly-, -Ala-, -His- and -Tyr-.\",\n \"Particularly suitable peptide sequence tags are described in EP 0 282 042B, and a preferred example is a hexa His tag.\",\n \"In one embodiment of the method of the invention, step (iii) is effected using a further antibody or a binding fragment thereof, which is specific for the peptide of amino acid sequence including SEQ ID NO 1.The said further antibody may be raised using conventional techniques to the peptide (7) which includes an amino acid of SEQ ID NO 1.This method is illustrated diagrammatically in FIG.\",\n \"2.The said further antibody is an anti-fusion antibody (8), which may be immobilised on a column, magnetic bead (9) or pipette tip, for example using a secondary antibody which is suitably an anti-species antibody (10) or other methods described in the literature, such as using an antibody binding protein such as Protein A, Protein G or Protein L, bound to the bead (9).\",\n \"This approach is highly suited to automation and to the isolation of large numbers, but small quantities, of novel fusion proteins in parallel.\",\n \"After separation from the cell lysate residue, the bound fusion protein (4) can subsequently be eluted by increasing the pH from 7.0 to 9.0.In an alternative embodiment, the fusion protein is isolated using a separation material which has some affinity for biotin but which releases the biotin fairly readily.\",\n \"Suitably the separation material is a modified version of avidin or streptavidin, which has lower affinity for biotin than native avidin or streptavidin.\",\n \"A particular example of such a material is a modified version of avidin marketed as CaptAvidin™ by by Molecular Probes (Eugene, Oreg., USA).\",\n \"In this embodiment, the fusion protein is isolated from the cellular debris, detergents and salts etc from the culture medium, by lowering the pH of the cell lysis mixture to pH 6.0 followed by affinity purification with CaptAvidin™ attached to (a) magnetic beads or (b) pipette tips using conventional methods.\",\n \"Bound fusion protein may then be eluted from the magnetic beads or mini columns by subsequently increasing the pH from 6.0 to 9.5.Prior to the step (iv), it is preferable to confirm the identity of the expressed fusion protein.\",\n \"In one particular technique, a very low volume (10 μl) of the isolated fusion protein is removed from the microtitre plate.\",\n \"The sample is digested by trypsin (using methods well known in the art).\",\n \"The resultant peptide extract is desalted and concentrated using a ZipTip™ (Millipore, Mass., USA) or equivalent, before analysis via mass spectrometry.\",\n \"Since the sequence of the fusion protein is known, identification by MALDI spectrometry to identify the peptides is usually sufficient to confirm the identity of the fusion protein.\",\n \"This technique is widely used in protein research and is summarised by T. Rabilloud (Editor) Proteome Research: 2D gel electrophoresis and identification methods.\",\n \"Furthermore, this technique can be automated and there are a number of commercially available systems from companies including Amersham Pharmacia Biotech, Bio Rad, AbiMed and Genomic Solutions (WO074852A1) that will perform this function.\",\n \"Similarly, prior to step (iv), it is preferable that the concentration of each expressed protein should be normalised where possible to eliminate variation between elements.\",\n \"Large variations in protein density cause difficulties in interpreting the data derived from such arrays (see Ekins, Clinical Chemistry (1998) 44:9 2015-2030, U.S. Pat.\",\n \"No 5,807,755 and U.S. Pat.\",\n \"No.\",\n \"5,432,099 for a detailed discussion on the quantitative aspects of protein immunoassays and protein arrays and definitions of assay sensitivity).\",\n \"Protein normalisation can be achieved by either determining the total protein concentration and or by including internal controls in the protocols.\",\n \"In a particularly preferred embodiment of the method of the invention, the fusion tag is used as an internal control and is detected by an antibody with a high affinity for the peptide of amino acid sequence which includes SEQ ID NO 1 within the fusion protein.\",\n \"Alternatively, the fusion protein can be expressed with a further peptide sequence tag and this can be used as an internal control.\",\n \"Such a tag may be the said further peptide sequence tag such as a hexa His tag as discussed above, which is expressed as part of the biotinylated fusion protein.\",\n \"The tagged version of this fusion protein may be detected through the use of an appropriate antibody such as an anti His tag antibody.\",\n \"This can be done by performing a classic immunoassay sandwich simultaneously with, or during, a subsequent analysis of a biological sample using the array.\",\n \"Using the antibody to the biotinylated fusion peptide, it is possible to determine the content of the fusion protein per μl and as a ratio of total protein present.\",\n \"The methodology may be performed using a sheep polyclonal primary antibody and secondary antibody sandwich in which the secondary antibody is conjugated with fluorescent dye (e.g.\",\n \"goat anti-mouse antibody conjugated to Alexa 488, Molecular Probes, Eugene, USA).\",\n \"The fluorescent dye used is spectrally distinct from any used with the secondary antibody for the biological sample.\",\n \"Both processes have been optimised for automation.\",\n \"The avidin or steptavidin coated non-porous support used in step (iv) of the method of the invention is suitably a glass or plastics material.\",\n \"Such supports are well suited to the production of small concentrated arrays.\",\n \"This is important, since biological samples are generally very limited in volume, and thus very valuable.\",\n \"A minimal surface area containing the targets is required for protein arrays, while still enabling the ability to achieve the required sensitivity of the assay is desirable.\",\n \"In addition, high density of either antigen or antibody in the array produces better signal to noise ratios when used in an assay.\",\n \"Furthermore, as compared to supports with porous surfaces including membranes, non-porous supports are more physically robust, are well suited to automation and have a lower background when imaged on fluorescent scanners.\",\n \"These may be coated with avidin or streptavidin using conventional methods.\",\n \"For example, the immobilisation of streptavidin to non-porous surfaces such as polystyrene multi-well plates is well known in the art.\",\n \"In its most basic form, a solution of streptavidin is left in contact with the surface for some hours.\",\n \"Un-bound protein is then removed by washing and the residual active moieties on the plastic surface blocked with BSA or an equivalent.\",\n \"Although this approach may be passive, it is effective.\",\n \"The non-covalent binding of streptavidin to polystyrene or nitrocellulose surfaces appears to be highly stable and resistant to elevated temperatures and high concentrations of chaotropic reagents, as described in WO98/37236.Avidin can be chemically attached to glass using the N-hydroxysuccinamide active ester of avidin as described by Manning, et al.\",\n \"Biochemistry 16: 1364-1370 (1977) and can be attached to nylon via carbodiimide based coupling methods as described by Jasiewicz, et al.\",\n \"Exp.\",\n \"Cell Res.\",\n \"100: 213-217 (1976).\",\n \"In another method, high molecular weight compounds such as biotin-N-hydroxy-succinimide ester, N-biotinyl-6-aminocaproyl-N-hydroxysulfosuccinimide ester, sulfosuccinimidyl-2-(biotinamido)ethyl-1,3-dithiopropionate were biotinylated and used to coat a suitable surface.\",\n \"Avidin or streptavidin was then coated in a second layer and was retained through binding the biotin linker attached to the high molecular weight compound as described in EP0620438.Attachment of streptavidin via a layer of biotin on the support surface was further developed in WO98/59243, which describes how biotin can be attached to a surface by chemical means or by light activation at 365 nm.\",\n \"The benefit that this provides is that regions of the surface can be masked.\",\n \"The elegance of these approaches is that biotin can be covalently bound to glass surfaces and will, in turn, non-covalently bind streptavidin only in those areas of the support that have been treated.\",\n \"This enables patterns of streptavidin “acceptor” protein on the support to be manufactured if required.\",\n \"In a preferred embodiment of the invention however, the entire surface of the non-porous support is coated with avidin or streptavidin, and then areas which are not required for binding are blocked, for example by addition of bovine serum albumin (BSA).\",\n \"In this way, any non-specific interaction of fusion protein with the support is reduced.\",\n \"Proteins that have been applied directly onto glass or plastic surfaces become non-covalently bound through interactions with charged groups on the solid surface the active moieties of the solid surface (typically silanol groups in glass or charged surface residues on polystyrene).\",\n \"Such non-specific adsorption of antigens or antibodies onto the surfaces of glass and plastic significantly reduces their antigenicity and antigen binding capacity respectively.\",\n \"The avidin or streptavidin layer therefore fulfils a dual role of firstly attaching the biotinylated fusion protein (non-biotinylated proteins that co-purify do not bind enabling a further purification step) and secondly, the dense layer of streptavidin shields the biotinylated fusion protein from undesirable non-specific interactions with the support surface.\",\n \"When the fusion protein is applied to the avidin or streptavidin coating on the support in step (iv), very tight but non-covalent bonding occurs.\",\n \"Preferably, the non-porous support is coated with streptavidin.\",\n \"Biotin attachment to streptavidin is multivalent, providing a binding of very high capacity when compared to that of the antigen bound directly to the support surface.\",\n \"Once the proteins have been applied to form the array, the bonding is strong enough to withstand extensive and stringent washing without appreciable loss of fusion protein.\",\n \"This is illustrated in FIG.\",\n \"3.Biotinylated fusion protein (4) is attached to the surface of the array support (12) via tight, non-covalent interaction with streptavidin (14).\",\n \"In the preferred example, streptavidin (14) is covalently bound to the support material.\",\n \"Sites on the array support material to which no streptavidin molecules are bound, are blocked by BSA or other surface modifiers (13).\",\n \"Fusion proteins bind the steptavidin (14) via the biotin label (7) on the fusion peptide (6).\",\n \"Furthermore, the avidin or streptavidin layer, whether attached directly to the surface of the support or via a biotinylated linker, is highly stable, is capable of being stored dry and can be heated and or treated with aggressive reagents without apparent loss of function (unlike most antigens and antibodies).\",\n \"Further “acceptor” layers can be constructed on top of the foundation of the streptavidin layer if required.\",\n \"These may comprise other antibody binding proteins known in the art.\",\n \"The array will have the advantage of using the concentrating effect of the streptavidin, which has multivalent sites for biotin attachment.\",\n \"This enables four times the biotin interaction with both antigen arrays and antibody arrays.\",\n \"This allows higher densities of either antigen or antibody in each element of the array which in turn means that more elements can be assembled per mm2 while achieving the same signal (see U.S. Pat.\",\n \"No.\",\n \"5,807,755 and U.S. Pat.\",\n \"No.\",\n \"5,432,099 for a discussion of the quantitative aspects) and less surface area of the solid support is utilised.\",\n \"The advantage is that less biological sample is thus required.\",\n \"Where the array is to consist of antigens arrayed on a microscope format, it suitably contains a large number of these, for example from 3-10,000 different fusion proteins.\",\n \"These may be generated or obtained from various sources, depending upon the intended nature and target for the analysis to be conducted using the array.\",\n \"Preferably, however, each protein will be expressed in the form of two fusion proteins, one with the peptide including SEQ ID NO 1 attached to the C-terminal, and one with the peptide including SEQ ID NO 1 attached to the N-terminal.\",\n \"In this way, the relative antigenicities of each version of the fusion protein to a complementary antibody can be assessed.\",\n \"The uses of antibody binding proteins such as Proteins A, G and L have been extensively reported.\",\n \"The binding of such proteins to antibodies is sufficiently tight to enable use in separation and detection techniques.\",\n \"These proteins are known to bind to the conserved regions of various classes of antibody.\",\n \"When the method of the invention is used to produce antibody arrays, antibody attachment is achieved by capture of the antibody via use of a layer of antibody binding proteins fused to a peptide which comprises SEQ ID NO 1.This layer preferably comprises of a mixture of biotinylated tagged Proteins A, G and L, and more preferably, some of which are fused at the C-terminal end, and some of which are labelled at the N-terminal end.\",\n \"By creating a mixture of antibody-binding proteins, a universal acceptor is created enabling the attachment of virtually any antibody, polyclonal, monoclonal, full-chain fragments, single chain antibodies and phage with antibody activity into which a Protein A, G or L site is present or has been engineered.\",\n \"Any antibody can be incorporated into the array without the need to pre-process or modify the antibody.\",\n \"When such antibody binding proteins are applied to an avidin or streptavidin coated surface in step (iv) a very high density of bound protein (biotinylated tagged protein binding to multivalent streptavidin) results.\",\n \"This method has the advantage that only one amino acid residue is biotinylated, and this is part of the fusion peptide, leaving the antibody binding sites on the lectins available.\",\n \"These antibody binding proteins effectively create a second layer on the solid surface.\",\n \"This layer comprising of bound fusion tagged Proteins A, G and L, acts as a universal acceptor surface for any antibody (FIG.\",\n \"6) without the need for direct biotinylation of the antibody.\",\n \"This saves time, antibody and eliminates the possible degradation of the antibody's binding to its corresponding antigen.\",\n \"In a preferred embodiment, a molar excess of antibodies is pre-mixed with biotinylated peptide antibody-binding protein (e.g.\",\n \"Proteins A, G or L) fusion and incubated for up to 15 minutes.\",\n \"This antibody-antibody binding protein mixture is then applied directly to the streptavidin covered array support.\",\n \"Alternatively, the biotinylated antibody binding protein fusion may first be applied to the streptavidin covered array support.\",\n \"Individual antibodies are then applied to the surface of the coated support to form an array.\",\n \"The array produced by either method comprises very discrete spots with minimal observable diffusion, leading to a good array for assay purposes.\",\n \"The array obtained using the method of the invention is suitably used in methods for detecting binding between antigens and antibodies.\",\n \"Thus in a second aspect, the invention provides a method of detecting binding between an antibody and an antigen, said method comprising the steps of (vi) applying to the array obtained using a method of the first aspect a sample which contains or is suspected of containing an antibody in the case of an array of step (v)(a), or an antigen in the case of the array of step (v)(b); and (vii) detecting bound antibody or antigen on the support.\",\n \"Steps (vi) and (vii) of the method of the invention are suitably carried out in a conventional manner, using well known immunlogical techniques such as ELISAs, including sandwich ELISAs using labelled and in particular fluorescently labelled antibodies.\",\n \"This is illustrated in the case of an antigen array in FIG.\",\n \"4.Antigens (4) bound to the array substrate (12) via streptavidin (14) are detected with a suitable primary antibody (15).\",\n \"The signal is amplified using a suitable secondary antibody (16) conjugated to a label (17).\",\n \"The label in the preferred embodiment is a fluorescent dye, such as Alexa 488, but may be any number of other types of label that are known in the art.\",\n \"Suitably the protein array continues to be monitored for quality and in particular the density of the protein during use of protein analysis devices.\",\n \"This is achieved in accordance with a preferred embodiment of the invention by using the peptide which comprises SEQ ID NO 1 or the further peptide sequence tag such as the hexa His Tag mentioned above, or any other suitable tag which performs this function as an internal standard, in a manner similar to that described above for pre-array protein normalisation.\",\n \"Once the antigens from numerous protein preparations have been arrayed onto the support surface, the array can then be used to assess antibody quality (see WO 99/39210) or can be used to determine antibody titre in serum samples (Joos et al Electrophoresis 2000, 21, 2641-2650).\",\n \"In these instances, the relative amounts of protein between the different elements in an array can be determined by adding an internal standard to the primary sample.\",\n \"The internal standard that is preferred is a sheep polyclonal antibody raised against the fusion peptide.\",\n \"This is spiked into the primary antibody solution (either antibody or serum) and is detected by an anti-sheep secondary antibody conjugated with a suitable fluorescent dye that is spectrally distinct from the labelled secondary antibody to the primary sample.\",\n \"A two-colour image is generated using a commercially available slide imager and the signal for each element is normalised to the signal resulting from the fusion protein.\",\n \"Combined with pre-array protein content normalisation, arrays of considerable consistency can be generated.\",\n \"Preferably at least some and most preferably all of the steps of the process described above are operated automatically to increase throughput and reduce labour time and costs.\",\n \"Creating antigen arrays with many novel proteins means proteins must be attached with the minimum number of steps if the process is to be viable.\",\n \"The use of a peptide which is readily and specifically biotinylated and which can act not only as a binding protein for assay purposes, but also as a purification means and an internal control for monitoring quality, provides just such a method.\",\n \"The method of the invention allows diverse collections of proteins to be attached with universal procedures, a minimum number of steps and maximum predictability of orientation.\",\n \"The method is suitable for operation on a large-scale, for example in high-throughput screening.\",\n \"In a third aspect the invention provides a protein array on a non-porous support, obtained using the method of the first aspect of the invention.\",\n \"Some elements used in the above-described methods are novel and therefore form further aspects of the invention.\",\n \"In particular, in a fourth aspect, the invention provides a fusion protein comprising an antibody binding protein fused at the N— or C-terminus to a peptide of 13 to 50 amino acids which comprises SEQ ID NO 1, such as a peptide of SEQ ID NO 2.In particular, the antibody binding protein is Protein A, G or L and preferably a mixture thereof.\",\n \"The fusion protein may additionally comprise a further peptide sequence tag such as the hexa His tag or another suitable sequence tag, which are known to those skilled in the art as discussed above.\",\n \"Such sequence tag may be located at the N or the C-terminus of the antigen or antibody binding protein.\",\n \"It is, however, preferably located at the opposite end of the antigen or antibody binding protein to which the amino acid sequence of SEQ ID NO 1 is fused.\",\n \"Where it is located at the same terminal region as SEQ ID NO 1, the sequence tag is fused to the free end of SEQ ID NO 1.A fifth aspect of the invention comprises a nucleic acid sequence which encodes the fusion protein of the fourth aspects.\",\n \"In particular in this case, the sequence which encodes the peptide is suitably of SEQ ID NO 9.\",\n \"(SEQ ID NO 9) GGCCTGAACGACATCTTCGAGGCTCAGAAAATCGAATGGCACGAA DESCRIPTION OF THE FIGURES FIG.\",\n \"1 illustrates diagrammatically the expression of a protein which is either an antigen or an antibody binding protein such as Protein A, G or L, in a form in which it can be used in the method of the invention.\",\n \"FIG.\",\n \"2 illustrates diagrammatically the isolation of fusion protein from cellular debris using an anti-tag antibody in accordance with an embodiment of the invention.\",\n \"FIG.\",\n \"3 illustrates diagrammatically the attachment of expressed fusion protein to a support surface coated with streptavidin in accordance with an embodiment of the invention.\",\n \"FIG.\",\n \"4 illustrates diagrammatically the detection of bound antigen with classic ELISA sandwich with the secondary conjugated to a fluorescent marker such as Alexa 488 in accordance with an embodiment of the invention.\",\n \"FIG.\",\n \"5 shows the results of a series of experiments in which a fusion protein comprising of GST fused to the fusion peptide was arrayed onto a streptavidin-coated microscope slide at several concentrations, the lowest in the above example being equivelent to 500 pg/spot.\",\n \"Panel (a) shows the image produced by a commercially available scanner using an excitation wavelength of 485 nm and an emission wavelength of 520 nm.\",\n \"In this example, the secondary antibody was conjugated to Alexa 488 as described in the text.\",\n \"Panel (b) shows an inverted image of (a) for ease of viewing.\",\n \"Pannel (c) shows an enlarged area of the array support with fusion protein at 500 pg/spot.\",\n \"The signal: noise ratio for these spots indicates that detection limits (signal:noise ratios of 3:1) would give a detection limit in the order of 10-50 pg protein per feature.\",\n \"FIG.\",\n \"6 (a) illustrates diagrammatically how solutions of Protein A, G and L (18) expressed as both C— and N-terminal fusion proteins were incubated with the streptavidin-coated slide (12).\",\n \"(a) Solutions of Proteins A, G and L (18) expressed as both C— and N-terminal fusion proteins were incubated with the streptavidin-coated slide (12).\",\n \"Antibodies (19) can be attached to the slide in highly discrete spots by arraying with a solid pin device or similar.\",\n \"The antibodies bind to the divalent protein binding proteins (Proteins A, G or L) at high densities.\",\n \"This was exemplified by binding a goat anti-mouse Ab conjugated to Alexa 488 and the image was acquired by scanning the slide with an emission wavelength of 485 nm and an excitation wavelength of 520 nm (inset panel (b)) FIG.\",\n \"7 shows the structure of the pAN-4 DNA vector obtainable from Avidity Inc. in which the S-D box (ASGGA) is shown in bold type, the start methionine codon is shown in italics and underlined, the sequence encoding the peptide of SEQ ID NO 2 is underlined, the ampicillin resistance bla is shown in bold type and the laclq is shown bold and underlined; FIG.\",\n \"8 shows the structure of the pAN-5 DNA vector obtainable from Avidity Inc. with annotations similar to those used in FIG.\",\n \"7; FIG.\",\n \"9 shows the structure of the pAN-6 DNA vector obtainable from Avidity Inc. with annotations similar to those used in FIG.\",\n \"7; FIG.\",\n \"10 shows the structure of the pAC-4 DNA vector obtainable from Avidity Inc. with annotations similar to those used in FIG.\",\n \"7; FIG.\",\n \"11 shows the structure of the pAC-5 DNA vector obtainable from Avidity Inc. with annotations similar to those used in FIG.\",\n \"7; FIG.\",\n \"12 shows the structure of the pAC-6 DNA vector obtainable from Avidity Inc. with annotations similar to those used in FIG.\",\n \"7.FIG.\",\n \"13 illustrates diagrammatically the expression of a protein, which is either an antigen, or an antibody binding protein such as Protein A, G or L, in a form in which it can be used in the method of the invention wherein the two alternative positions of the second peptide sequence tag are shown.\",\n \"DETAILED DESCRIPTION OF THE INVENTION Step 1: Cloning All genes expressed were cloned from cDNA preparations directly into each of the pAN and pAC series of vectors (Avidity Inc, USA).\",\n \"These were used to express N-terminal and C-terminal fusion proteins respectively.\",\n \"The fusion peptide sequence used was SEQ ID NO 2 shown above.\",\n \"The insert sequences were confirmed by DNA sequencing performed on 377 (PE Corporation Inc) and MagaBase (Amersham Pharamcia Biotech) instruments using the manufacturer's methodologies.\",\n \"Step 2: Expression All fusion proteins were expressed under the control of the tightly repressed Trc promoter and is IPTG-inducible.\",\n \"All proteins were expressed in strain AVB100 (Avidity Inc, Colo., USA), an E. coli K12 strain [MC1061 ara D139 delta(ara-leu)7696 delta(lac)174 galU galK hsdR2(rK-mK+) mcrBl rpsL(Strr)] with a birA gene stably integrated into the chromosome.\",\n \"Over expression of the BirA protein was accomplished by induction with L-arabinose.\",\n \"The stably integrated birA gene does not require antibiotics to be maintained, and use of AVB100 with IPTG-inducible vectors such as pAC and pAN, vectors (Avidity Inc, USA) allowed independent control over the expressed gene of interest and the BirA levels.\",\n \"Strain AVB99 (Avidity Inc) was also used and is an E. coli strain (XL1-Blue) containing a pACYC184 plasmid with an IPTG-inducible birA gene to overexpress biotin ligase (pBirAcm).\",\n \"Strain AVB101 (Avidity Inc) was also used and is an E. coli B strain (hsdR, lon11, sul Al), containing a pACYC184 plasmid with an IPTG-inducible birA gene to overexpress biotin ligase (pBirAcm).\",\n \"Expression of both biotin ligase and the fusion protein was induced with IPTG (1 mM).\",\n \"Biotin was added at the time of induction to a concentration of 50 μM.\",\n \"Step 3: Purification Biotinylated fusion proteins were isolated by two separate methods.\",\n \"These methods can either be used as alternates or were combined as a two-stage process where ultra-pure preparations were required.\",\n \"a) Purification Using Anti-Fusion Peptide Antibodies A partially purified mouse monoclonal antibody to the C-terminus fusion peptide was available and polyclonal antibodies to the C— and N-terminal fusion proteins were raised in rabbit.\",\n \"i) In one of the methodologies, the anti C-terminal mouse monoclonal was attached directly to magnetic beads using 2.4 micron magnetic beads with a tosylated activated surface (Dynal Biotech ASA, Norway) as follows: Coating procedure.\",\n \"Dynabeads M-280 Tosylactivated were resuspended by pipetting and vortexing for approximately 1 min and were immediately pipetted into the reaction tube.\",\n \"Supernatant was removed from the beads using a magnet (Dynal MPC) to separate the beads from solution.\",\n \"The supernatant was removed, leaving beads undisturbed.\",\n \"The beads were resuspended in an ample volume of 0.I M Na-phosphate buffer pH 7.4 and mixed gently for 2 min.\",\n \"After using the magnet again and pipetting off the supernatant, the washed beads were resuspended in the same volume of 0.1 M Na-phosphate buffer pH 7.4 to the required concentration.\",\n \"The appropriate antibody was dialysed into 0.1 M Na-phosphate buffer pH 7.4.The amount of antibody was approximately 3 μg antibody per 1 Dynabeads (approximately 20 μg/mg) and the beads were resuspended by vortexing for 1 min.\",\n \"The mixture was incubated for 16-24 h at 37° C. with slow tilt rotation.\",\n \"After incubation, the magnet was used to separate the magnetic beads for 1-4 minutes and the supernatant was removed The coated beads were washed four times (twice in ×1 PBS pH 7.4 [phosphate buffered saline] with 0.1% [w/v] BSA for 5 minutes at 4° C.) once with 0.2 M Tris-HCl pH 8.5 with 0.1% (w/v) BSA for 24 hours at 20° C. or for 4 hours at 37° C. (Tris blocks free tosyl-groups) and finally once in ×1 PBS, pH 7.4 with 0.1% [w/v] BSA for 5 minutes at 4° C. The Dynabeads M-280 Tosylactivated are thereby coated with the antibody.\",\n \"The cells expressing the fusion protein of interest were lysed for 15 minutes in ice-cold ×1 PBS, pH 7.4 with 1% NP-40 and protease inhibitors, after which the lysate was centrifuged at 2,000×g for 3 minutes.\",\n \"The lysate was pre-cleared by incubation of the ice-old lysate (in 1.5 ml Eppendorf tubes) for 2 hours with Dynabeads pre-coated with the appropriate antibody (0.5 mg Dynabeads pr.\",\n \"lysate from 1×106 cells).\",\n \"The Dynabeads were washed 3 times in 1.5 ml ice-cold PBS/1% NP40 by using a Dynal Magnetic Particle Concentrator to collect the beads at the wall after each washing step.\",\n \"The fusion protein-antibody magnetic bead complex was disrupted by adjusting the pH to above 9.0.Supernatant was separated from the magnetic beads with the Magnetic Particle Concentrator and assayed for total protein concentration, concentration of fusion peptide and the protein was identified by mass spectrometry using a PerSeptive Voyager MADLI (see below).\",\n \"ii) In the second methodology, the antibody was attached indirectly to Dynal magnetic beads via Protein A and Protein G previously immobilised onto the surface of the bead by the manufacturer.\",\n \"A mixture of Dynabeads-Protein G and Dynabeads-Protein A were resupended by vortexing for 1-2 minutes.\",\n \"The supernatant was removed from the beads using a magnetic workstation as described above.\",\n \"0.5 ml 0.1 M Na-phosphate buffer pH 7.0 containing 0.01% Tween 20 and 0.1% (w/v) BSA were added and the wash procedure repeated three times.\",\n \"The antibody was added to the washed Dynabeads and incubated with gentle mixing for 10-40 minutes.\",\n \"The supernatant was removed using the magnetic workstation.\",\n \"The beads were twice resuspended in 0.5 ml 0.1 M Na-phosphate buffer pH 7.0 containing 0.01% Tween 20 and 0.1% (w/v) BSA for protein stability.\",\n \"Supernatant was removed and the beads added to the lysate mixture as prepared above.\",\n \"Binding of the fusion protein was performed at 2-8° C. for 10 minutes to 1 hour.\",\n \"Approximately 25 μg target protein per μl of the initial Dynabeads Protein G volume was used to assure an excess of protein.\",\n \"Incubation was performed while tilting and rotating the tube with incubation times as low as 10 minutes.\",\n \"Supernatant containing detergents and cell lysate was removed from the fusion-protein-Ig Dynabeads-Protein G complex using the magnetic workstation and washed 3 times using ×1 PBS, pH 7.4 with 0.01% Tween 20.Bound fusion protein was best eluted from the fusion-protein-Ig Dynabeads-Protein G/A complexes by adjusting the pH to above 9.0 and removing the supernatant containing the now purified fusion protein.\",\n \"Supernatant was separated from the magnetic beads with the Magnetic Particle Concentrator and assayed for total protein concentration, concentration of fusion peptide and the protein was identified by mass spectrometry using a PerSeptive Voyager MADLI (see below).\",\n \"iii) In another example, Proteins A, G and L mixtures were immobilised on to suitably prepared pipette tips.\",\n \"The antibody was incubated with the pipette tips in 50 mM Tris-HCl buffer, pH 8.0 containing 0.01% Tween 20 and 0.1% (w/v) BSA for 60 minutes at room temperature.\",\n \"The coated pipette tips were then rinsed with 3 pipette volumes of 50 mM Tris-HCl buffer, pH 8.0 containing 0.01% Tween 20 and 0.1% (w/v) BSA.\",\n \"200μl cell lysate was aspirated from the bottom of the tip either by hand or with a robotic workstation several times to ensure the extraction of the biotinylated fusion protein.\",\n \"The cell lysate was discarded.\",\n \"The pipette tips were rinsed with three volumes of 10 mM Tris-HCl buffer, pH 8.0 containing 0.01% Tween 20 and 0.1% (w/v) BSA.\",\n \"Bound fusion protein was eluted in half a pipette volume of 50 mM sodium bicarbonate-HCl buffer, pH 10.0 containing 0.01% Tween 20 by gently aspirating this aliquot up through the bottom of the pipette tip.\",\n \"The resulting solution containing the fusion protein was assayed as described below.\",\n \"iv) In another preferred methodology an alternative version of the biotinylated fusion protein was constructed with the addition of a hexa His tag.\",\n \"The hexa His fusion peptide is often used as a standard purification procedure and is well known to those skilled in the art.\",\n \"Typically, cells were lysed in 5 ml buffer per gram wet weight of cells.\",\n \"The lysis buffer comprised: ×1 NBB (20 mM Tris CL, 100 mM NaCl, 5 mM Imidazol, pH 8.0) with 1 in 100 volume of 10 mg/ml lysozyme, 1 in 100 volume protease inhibitor cocktail (Calbiochem protease inhibitopr cocktail set 3), 10 mM beta mercaptoethanol, supplemented with a ×1 detergent cocktail supplied by Novagen (Madison, USA).\",\n \"The cells were lysed for 15 minutes at 30-37° C. Cellular proteins were denatured by adding Urea to a final concentration of 6M and 2M thiourea The solution was clarified by passing through a 0.22 micron filter, and then applied directly onto nickel agarose matrix (NTA supplied by Qiagen, Germany).\",\n \"Proteins were incubated with the nickel agarose beads for 15 minutes and the non-binding protein removed by centrifugation.\",\n \"The beads were washed three times in 10 volumes of the lysis buffer supplemented with 6M Urea and 2M Urea.\",\n \"After the final wash, 50% of the wash buffer was removed and then diluted with a 20 mM Tris HCl, 100 mM NaCl, pH 8.0 buffer containing 10 mM beta mercaptoethanol.\",\n \"This step was repeated three times.\",\n \"Finally the beads were washed with 10 volumes of buffer, the composition of which was 20 mM Tris HCl, 100 mM NaCl, pH 8.0 buffer (without urea/thiourea).\",\n \"The proteins were eluted several aliquots of buffer (20 mM Tris HCl, 100 mM NaCl, pH 8.0 buffer), supplemented with various concentrations of imidazole.\",\n \"The typical concentration range of imidazole used to eluted the bound protein was between 20 mM to 500 mM.\",\n \"The fractions containing the eluted protein were pooled.\",\n \"b) Purification Using CaptAvidin™ (Molecular Probes Inc. Oregon.\",\n \"USA) In another experiment, the biotinylated fusion protein was isolated using a novel form of streptavidin marketed as CaptAvidin™ (Molecular Probes, Oregon, USA) immobilised to a suitable surface.\",\n \"In this modified form of streptavidin, the tyrosine residue in the biotin binding sites is nitrated, thereby reducing the very strong non-covalent bond with a Ka of 1015M−1 to a Ka of 109M−1.The association between biotin and CaptAvidin™ can therefore be disrupted by raising the pH to between 9-10 as described below: i) In one preferred embodiment, CaptAvidin™ protein was attached to tosylated magnetic beads (Dynal Biotech ASA, Norway) and was washed and prepared as described above.\",\n \"The CaptAvidin™ coated beads were washed three times in 50 mM citrate phoasphate buffer, pH 4.0 containing 0.01% Tween 20 and 0.1% (w/v) BSA and the supernatant was discarded.\",\n \"The cell lysate mixture was prepared as described above and the pH adjusted to 5.0.CaptAvidin™ coated beads were added at a ratio of 0.5 mg Dynabeads per lysate from 1×106 cells.\",\n \"The solution was incubated with gentle agitation for 10-60 minutes.\",\n \"The supernatant was removed from the magnetic beads using a magnetic workstation (Dyanl Biotech ASA, Norway) and washed with three aliquots of 10 mM Tris-HCL buffer, pH 8.0 containing 0.01% Tween 20, discarding the supernatant.\",\n \"The biotinylated fusion protein is detached from the CaptAvidin™ coated magnetic beads by adding an aliquot of 50 mM sodium bicarbonate-HCl buffer, pH 10.0 containing 0.01% Tween 20 and gently agitating the slurry for 15 minutes at room temperature.\",\n \"The magnetic beads were removed using the magnetic workstation and the supernatant containing the biotinylated fusion protein was retained.\",\n \"ii) In another example, the magnetic beads were replaced by creating mini columns of CaptAvidin™ conjugated to agarose beads (Molecular Probes Inc, Oregon, USA) mixed with an equal volume of Sepharose® CL-4B agarose (Amersham Pharmacia Biotech Ltd, UK) to increase the bed volume with mini columns made by pouring the slurry into pipette tips in 50 mM citrate phosphate buffer, pH 4.0 containing 0.01% Tween 20.Biotinylated fusion protein was separated from cell lysate mixture by affinity chromatography.\",\n \"Unbound material is eluted from the column with 10 column volumes of 10 mM Tris-HCl buffer, pH 8.0 containing 0.01% Tween 20.Biotinylated fusion protein was eluted from the column in two column volumes of 50 mM sodium bicarbonate-HCl buffer, pH 10.0 containing 0.01% Tween 20.iii) In yet another experiment, the CaptAvidin™ agarose beads were immobilised into a pipette tip and fusion protein binding and elution was performed as described above.\",\n \"Step 4: Protein Identification Expressed and purified fusion proteins were identified by peptide finger printing.\",\n \"Using methods as reviewed in Proteome Research (Edited by Rabilloud), the fusion protein was digested with trypsin, the resulting peptide solution was desalted and concentrated using a ZipTip™ (Millipore, Mass., USA) reverse phase column, diluted into matrix solution and applied to a target plated from a PerSeptive Voyager™ mass spectrometer and analysed by MADLI.\",\n \"The resulting spectra of peptide masses were compared with the anticipated peptide finger print for the protein using the ExPASy search algorithms (GeneBio AG, Switzerland) via their website (www.expasy.com).\",\n \"Step 5: Protein Assay (Normalisation) A 3-5 μl aliquot of the purified fusion protein was removed from the stock solution and assayed for total protein content using the BCA method in preference to Bradford assay due to the presence of detergents in the protein samples.\",\n \"The concentration of biotinylated fusion protein was determined by immunoassay as follows; A 3-5 μl aliquot of the purified fusion protein was removed from the stock solution and incubated in a black, streptavidin-coated microtitre plate (Beckton Dickenson, USA).\",\n \"The well was washed three times with 50 mM Tris-HCL buffer, pH 8.0 containing 0.01% Tween 20.The well was blocked using 1% (w/v) BSA in the same buffer for 30 minutes and then rinsed three times with 50 mM Tris-HCL buffer, pH 8.0 containing 0.01% Tween 20.The immobilised biotinylated fusion protein was incubated with either an anti N-terminal or anti C-terminal polyclonal antibody raised in rabbit diluted into 50 mM Tris-HCL buffer, pH 8.0 containing 0.01% Tween 20 and 0.1% (w/v) BSA.\",\n \"The well was rinsed three times with buffer and then probed with a anti-rabbit, mouse monoclonal conjugated to Alexa 488 (Molecular Probes Inc, Oregon, USA) and the signal measured with a PerkinElmer Flight fluorescence plate reader.\",\n \"A standard curve with known amounts of Glutathione S-transferase expressed using the expression system described in U.S. Pat.\",\n \"No.\",\n \"5,723,584, U.S. Pat.\",\n \"No.\",\n \"5,874,239 and U.S. Pat.\",\n \"No.\",\n \"5,932,433 was used for calibration in the range of 0.1-500 μg of fusion protein per well.\",\n \"Step 6: Manufacture of Protein Arrays a) Creation of Streptavidin Coated Microscope Slides Microscope slides coated with streptavidin were first imaged on a variety of commercially available slide readers using an excitation wavelength of 480 nm and and emission wavelength of 520 nm to assess the evenness of the coating.\",\n \"b) Manufacture of Antigen Arrays The streptavidin coated slides were rehydrated with ×1 phosphate buffered saline at pH 7.3.Purified biotinylated fusion proteins at a concentration of approximately 1 μg/μl were spotted onto the surface of the slide using a solid pin with a tip diameter of 100-150 microns (Biorobotics, Cambridge, UK) by hand and with a robotic system.\",\n \"The slide was incubated at room temperature in a humidity-controlled environment for 30 minutes.\",\n \"The slide was then typically washed with ×1 PBS, pH 7.3 containing 0.01% (v/v) Tween and then blocked by incubating the slide with 1% (w/v) BSA for 10 minutes.\",\n \"The slide was rinsed with ×1 PBS, pH 7.3 containing 0.01% (w/v) Tween 20 and then incubated with the primary antibody of choice diluted 1:400 in ×1 PBS, pH 7.3 containing 0.01% (w/v) Tween 20 and 0.1% (w/v) BSA, or a complex biological mixture of proteins containing immunoglobulins, e.g.\",\n \"diluted serum samples.\",\n \"The slide was then rinsed in ×1 PBS, pH 7.3 containing 0.01% (w/v) Tween and 0.1% (w/v) BSA and incubated with an appropriate secondary (for example mouse anti-human IgG monoclonal conjugated to Alexa 488 (Molecular Probes Inc) for the detection of immunoglobulins in serum, for example).\",\n \"The slides were then imaged at excitation/emission wavelengths of 480/520 nm, for the Alexa 488 conjugate, although one skilled in the art can appreciate that many such secondary Abs with a variety of labels (colorimetric, alternative fluorescent, radiolabelled or chemiluminescent) could be used in its place.\",\n \"An example of the results obtained is illustrated in FIG.\",\n \"5 hereinafter.\",\n \"c) Manufacture of Antibody Arrays Creation of a Universal Antibody Acceptor Layer Proteins A, G and L from Streptococcus aureus were cloned into the expression vectors pAN-4, pAN-5 or pAN-6, pAC4, pAC-5 and pAC-6) and were expressed and purified as described above, resulting in both C— and N-terminal fusion proteins which were biotinylated in vivo, again as described above.\",\n \"Streptavidin coated microscope slides were coated with a mixture of fusion proteins (both C— and N-terminal fusions) of Proteins A, G and L in ×1 PBS, pH 7.3 at a concentration of 1 mg/ml.\",\n \"The slides were incubated at room temperature for a minimum of 30 minutes in a humidity-controlled environment.\",\n \"The slides were washed with ×1 PBS, pH 7.3 containing 2 mM Sodium Azide and were stored in sealed containers in a moist atmosphere (to prevent drying) at 4° C. until required.\",\n \"Printing Antibody Arrays The universal antibody acceptor layer was used to attach a variety of different classes of antibodies and those phage molecules engineered to include a Protein A, G or L binding site.\",\n \"Antibody preparations are diluted in 1× PBS, pH 7.3 containing 0.01% Tween to a concentration of 0.2-10 mg/ml.\",\n \"The antibody solutions were applied to the universal antibody acceptor layer with solid pins with a tip diameter of between 100-150 microns (Biorobotics, Cambridge, UK) by hand or with a robotic system.\",\n \"The slides were then blocked with 1% BSA in xl PBS, pH 7.3 containing 0.01% Tween.\",\n \"Slides were rinsed with the ×1 PBS, pH 7.3 containing 0.01% Tween and 2 mM Sodium Azide and were stored in sealed containers in a moist atmosphere (to prevent drying) at 4° C. until required.\",\n \"Scanning as described above for antigen arrays produced the sort of results which are illustrated in FIG.\",\n \"6.Step 7: Labelling Complex Mixtures of Proteins with Fluorescent Dyes Typically, protein samples were prepared by solubilising them in a variety of buffers and detergents, depending on the biological sample.\",\n \"Many samples required aggressive solubilisation procedures requiring the use of non-ionic detergents and 8M urea, similar to those used in the preparation of proteins for the first dimension of 2D electrophoresis gels.\",\n \"For example, the solublization methodology involved homogenization of the sample into solution containing 4% CHAPS, 50 mM PBS, pH 7.6 with either 7 M urea and 2 M thiourea or 8 M urea Buffers containing primary amino groups such as TRIS and glycine inhibit the conjugation reaction and were therefore avoided.\",\n \"The presence of low concentrations (<2%) of biocides such as azide or thimerosal did not affect protein labelling.\",\n \"The solubilised protein was centrifuged at 10,000 g to remove cellular debris and non-solubilised material and the mixture was immediately labeled.\",\n \"Complex mixtures of proteins from biological samples were labelled with a fluorescent tag prior to incubation with the antibody array as prepared above.\",\n \"Clearly, those skilled in the art will recognise that other forms of labels can be applied to the technique such as radiolabelling, chemiluminescent and visual dyes.\",\n \"Further, other fluorecent dyes can also be applied to the process.\",\n \"One preferred embodiment is the use of Cy3 and Cy5 mono reactive dyes (Amersham Pharmacia Biotech Ltd, UK).\",\n \"Dye labelling of complex protein mixtures was unpredictable and had to be optimised for each type of biological sample.\",\n \"Specifically, the binding of dye molecules to proteins via residues with amine groups often reduced the antigenicity of certain proteins such that they were no longer recognised by a functional antibody.\",\n \"The manufacturer's recommended procedure is designed to label 1 mg protein to a final molar dye/protein (D/P) ratio between 4 and 12.This assumes an average protein molecular weight of 155,000 daltons.\",\n \"In the present invention, an average dye/protein ratio above 2-3 was found to interfere with the antibody-antigen reaction for many of the proteins studied.\",\n \"It was determined that the D/P ratios could be simply controlled by using different concentrations of protein and different buffer pH values.\",\n \"Altering the protein concentration and reaction pH changed the labelling efficiency of the reaction significantly.\",\n \"Optimal labelling occured at pH 9 and by reducing the pH to 7.6 reduced the dye/protein ratio to between 1-3.Higher protein concentrations increased labeling and so the control of protein concentration was also found to be critical.\",\n \"Solutions of up to 10 μg/μl of a single protein species gave dye/protein ratios of 10-14, so more appropriate concentrations were found to be 0.1-1.0 μg/μl.\",\n \"A typical method was as follows: complex protein mixtures prepared as described above, were diluted to several concentrations in ×1 PBS buffer, pH 7.6 containing 0.2% CHAPS to achieve an average protein species concentration of 1.0 μg/μl (total protein concentration was in the range of 50-100μg/μl) The protein solution was incubated at room temperature for 30 minutes with constant gentle agitation.\",\n \"Labeled protein must be separated from the excess, unconjugated dye prior to incubation with the antibody arrays.\",\n \"The manufacturer recommends separation from unbound protein by gel permeation, however, due to the presence of membrane-bound proteins with poor solubility this step was replaced by simply adding an excess of glycine to the solution to halt the reaction.\",\n \"The labeled protein solution was incubated for a further 15 minutes to ensure the removal of residual free dye.\",\n \"Labeled proteins were stored at 2-8° C. without further manipulation.\",\n \"Free dye was also removed using the method of Ünlü et al (1997) in which free dye was removed by overnight incubation with SM-2 beads (Bio-Rad, CA, USA).\",\n \"The final dye/protein (D/P) ratio was estimated as follows: a portion of the labeled protein solution was diluted so that the maximum absorbance was 0.5 to 1.5 AU.\",\n \"Molar concentrations of dye and protein were calculated.\",\n \"The extinction coefficient will vary for different proteins but is a reasonable average to use for complex mixtures.\",\n \"The ratio of the average number of dye molecules coupled to each protein molecule was calculated as follows: Cy5/Protein ratios were calculated using molar extinction coefficients of 250,000 M− 1 cm−1 at 650 nm for Cy5, and 170,000 N−1 cm−1 at 280 nm for the protein mixture.\",\n \"The calculation was corrected for the absorbance of the Cy5 dye at 280 nm (approximately 5% of the absorbance at 650 nm) as per the manufacturer's product data sheets.\",\n \"[Cy5 dye]=(A650)/250000, [protein]=[A280−(0.05×A650)]/170000, (D/P) final=[dye]/[protein], (D/P) final=[0.68× (A650)]/[A 280−(0.05×A650)).\",\n \"Cy3 /Protein ratios were calculated using molar extinction coefficients of 150,000 M−1 1 cm−1 at 552 nm for the Cy3 dye and 170000 M−1 cm−1 at 280 nm for the protein are used in this example.\",\n \"The calculation was corrected for the absorbance of the dye at 280 nm (approximately 8% of the absorbance at 552 nm).\",\n \"[Cy3 dye]=(A552)/150000, [antibody]=[A 280−(0.08×AS52)]/170000, (D/P) final=[dye]/[antibody], (D/P) final=[1.13×(A552)]/[A280−(0.08×A552)).\",\n \"Step 8: Determination of Protein Expression Using Antibody Arrays Cy3-labelled and Cy5-labelled proteins were mixed in equimolar amounts based on the Dye/protein ratios determined above.\",\n \"100 μl of the mixture was incubated with a antibody array that had previously been rinsed with several slide volumes of ×1 PBS, pH 7.6 containing 0.01% Tween.\",\n \"The labelled protein mixture was incubated at 30° C. for one hour in an automated slide processor subject to UK Patent Application GB 0028647.6 (unpublished).\",\n \"The slide was then rinsed with 10 slide volumes of ×1 PBS, pH 7.6 containing 0.01% Tween.\",\n \"The slides were dried by centrifugation and imaged immediately on a commercially available slide imager using the manufacturer's operating procedures.\",\n \"The Cy3 and Cy5 labelled protein ratios were analysed and normalised to a number of marker proteins such as actin and GAPDH.\",\n \"While this approach is suitable for similarly prepared tissues or other biological samples, care must be taken on the applicability of this normalisation strategy between different tissue types and other biological samples, since the total cell content of all proteins vary considerably from tissue to tissue.\",\n \"The potential of protein arrays has been discussed for many years and clearly is a much needed tool.\",\n \"The problems with expressing, purifying, assaying and in particular, attaching proteins to solid, non-porous surfaces have all proved difficult problems to solve.\",\n \"Through the novel exploitation of the vector technology described in patents U.S. Pat.\",\n \"No.\",\n \"5,723,584, U.S. Pat.\",\n \"No.\",\n \"5,874,239 and U.S. Pat.\",\n \"No.\",\n \"5,932,433, the present invention provides a method for the preparation of both antigen and antibody arrays that allow researchers to now apply these techniques with greater success.\",\n \"All references mentioned in the above specification are herein incorporated by reference.\",\n \"Other modifications of the present invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention.\",\n \"Although the invention has been described in connection with the specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments.\",\n \"Indeed, various modifications of the described modes for carrying out the invention, which are obvious to those skilled in the art, are intended to be within the scope of the following claims.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473882"}}},{"rowIdx":761,"cells":{"text":{"kind":"list like","value":[["Medicament dispenser","Apparatus for dispensing medication to a patient, said apparatus comprising: (a) an electronic medicament dispenser (2) with one or more data-transfer elements; (b) a docking station (6) with one or more corresponding data-transfer elements; and (c) a wireless communications module (10) for connecting said docking station to a public communications network, whereby data may be transmitted remotely between said medicament dispenser and a remote processing node via said public communications network."],["1.Apparatus for dispensing medication to a patient, said apparatus comprising: (a) an electronic medicament dispenser with one or more data-transfer elements; (b) a docking station with one or more corresponding data-transfer elements; and (c) a wireless communications module for connecting said docking station to a public communications network, whereby data may be transmitted remotely between said medicament dispenser and a remote processing node via said public communications network.","2.Apparatus according to claim 1, wherein said wireless communications module forms an integral part of said docking station.","3.Apparatus according to claim 1, wherein said data-transfer elements comprise electrical contacts.","4.Apparatus according to claim 1, wherein said docking station comprises a display for displaying data relating to said medicament dispenser.","5.Apparatus according to claim 1, wherein said medicament dispenser comprises a display, said medicament dispenser display being smaller in area than said docking station display.","6.Apparatus according to claim 1, wherein said medicament dispenser comprises means for authenticating data received from said docking station 7.Apparatus according to claim 1, wherein said docking station comprises means for initiating communications sessions with a network entity in response to docking of said dispenser therewith.","8.Apparatus according to claim 1, wherein said dispenser comprises a removable refill container.","9.Apparatus according to claim 8, wherein said docking station is arranged to transmit data relating to any conflict between said dispenser and said refill to a network entity via a communications link.","10.Apparatus according to claim 1, wherein said docking station is arranged to form a link via said wireless communications module with one or more of an appropriately configured personal computer workstation, a digital television set top box, a digital television and/or a home communications hub.","11.A method of controlling the dispensing medication to a patient, the method comprising: (a) providing an electronic medicament dispenser with one or more data-transfer elements; (b) providing a docking station with one or more corresponding data-transfer elements; and (c) providing a wireless communications module for connecting said docking station to a public communications network, (d) transmitting data remotely between said medicament dispenser and a remote processing node via said public communications network, using said docking station and said wireless communications module.","12.A method according to claim 11, comprising transmitting data from said medicament dispenser to said remote processing node.","13.A method according to claim 11, comprising transmitting data from said remote processing node to said medicament dispenser."],["The present invention relates to medicament dispensers for use in the dispensing of medicament, to medicament containers for use in same, and to systems and methods for the dispensing of medicament.","Lack of patient compliance with medical treatment regimens is known to be a problem.","According to the United States Food and Drug Administration (FDA), between 30 and 50 percent of patients fail to use medicines as prescribed, see “FDA Proposes Program to Give Patients Better Medication Information”.","The main problems are known to be taking improper dose, failing to take doses on time and ceasing treatment prematurely.","If this problem could be addressed, the quality of care offered to patients could be improved considerably.","Medical dispensers are well known for the dispensing of various kinds of medicament.","In their most simple form, they may comprise a container of tablets with a removable lid or blister packs of tablets.","Inhalation devices, such as metered dose inhalers (MDI) and dry powder inhalers are known for the delivery of medicament for the treatment of respiratory disorders.","Syringes, including needleless syringes are also known for the delivery of injectable medicament to a patient.","Reloadable medicament dispensers are known.","These typically comprise a housing defining a cavity and a medicament container, referred to herein as a refill, which is reversibly receivable thereby.","The housing and the medicament container may be sold separately or as a kit of parts.","Currently, medicament dispensers are primarily manually operable.","A patient actuates the dispenser in order to receive a dose of the medicament.","If a variation in the basic dose, e.g.","more than one dose, is required, the patient typically manually administers two doses.","It would be desirable to provide a way in which the patient is relieved of sole responsibility for the correct administration of doses of medicament, and more preferably the variation of such doses, within a treatment regimen.","Further desirable features relating to medicament dispensers include ease of use, portability and ease of manufacture.","PCT patent application no.","WO92/17231 describes a metered dose inhaler having a microelectronic assembly thereon.","The medicament container includes a set of electrically conducting strips which represent information about the medicament container in digital form.","The housing of the device includes electrical contact fingers which are contactable with the strips to enable reading of the information to a microelectronic memory on the housing.","Read/write communication would not be possible between the fingers and the reader and the significant advantages of the present invention would therefore not be achievable with this inhaler.","Furthermore, contact between the strips on the container and the electrical contact fingers is required which requires physical tailoring of the container to the housing, thereby limiting product design options.","This document also describes an embodiment in which an active (i.e.","powered) microelectronic element is attached to the container.","PCT Patent Application No.","WO 00/25720 describes a medication container that organises several vials or cassettes of different types of medication by securing the vials to a unitary lid.","A machine readable memory strip is affixed to each vial.","Each memory contains prescription information and medication implementation pertaining to the medication in the vial.","The unitary lid is equipped with sensors that read each memory strip and transmit the information to a processor in the lid.","The processor determines when each medication is to be taken and signals the patient to take the appropriate medication from the appropriate vial at the appropriate time.","The automated lid also contains a receiver for obtaining updated medication dosing information based on current laboratory tests or physical observations of the physician regarding the patient.","It is an object of the present invention to provide improvements in relation to systems in which electronic medicament dispensers are used in a course of treatment.","In accordance with one aspect of the present invention there is provided apparatus for dispensing medication to a patient, said apparatus comprising: (a) an electronic medicament dispenser with one or more data-transfer elements; (b) a docking station with one or more corresponding data-transfer elements; and (c) a wireless communications module for connecting said docking station to a public communications network, whereby data may be transmitted remotely between said medicament dispenser and a remote processing node via said public communications network.","By providing an electronic medicament dispenser with one or more data-transfer elements, and a docking station with one or more corresponding data-transfer elements, functionality which is not essential to the short-term functioning of the medicament dispenser may be placed in the docking station.","By providing a separate docking station, features which are not essential to the short-term functioning of a dispenser may be instead implemented in the docking station, thereby increasing the versatility of the system.","These features may include a data transfer feature.","The provision of a wireless communication module for connecting the docking station to a public communications network allows the functioning of the docking station to be augmented by resources provided in a remote processing node, whilst allowing the location of the docking station within the user's environment to be selected by the user.","In contrast, requiring the user to locate a docking station at a public data communications access point, such as a telephone access point, would in many cases prevent the frequent and correct usage of the device which is desired in a system in accordance with the present invention.","Further features and advantages of the present invention will become apparent from the following description of preferred embodiments of the invention.","Embodiments arranged in accordance with the invention will now be described, by way of example only, with reference to the accompanying drawings in which: FIG.","1 is a schematic illustration of a healthcare management system in accordance with an embodiment of the invention; FIG.","2 is a schematic illustration of patient apparatus in accordance with an embodiment of the invention; FIGS.","3 and 4 illustrate look-up tables used in embodiments of the invention; FIGS.","5 to 8 are flow diagrams illustrating procedures carried out in accordance with embodiments of the invention; and FIGS.","8 to 11 illustrate embodiments of patient apparatus in greater detail.","FIG.","1 illustrates a healthcare management system arranged in accordance with an embodiment of the invention.","The system includes a patient healthcare management system A, a healthcare service provider data management system B, a physician's data management system C, a pharmacy data management system D, and manufacturer data management system E. Patient healthcare management system A includes a medicament dispenser 2 which includes an outer housing which takes a refill 4, containing medicament, which is inserted into a receiving inlet 5 of the dispenser and securely fastened therein.","The refill 4 is removable therefrom once the refill is empty, or near to empty, to be replaced by a further refill.","The dispenser includes an alphanumeric display 28 for the display of functional data and one or more man machine interface elements, for example buttons 30, for the control of the functions of the dispenser by the patient.","The dispenser 2 is adapted for insertion, in a drop-in fashion, into a docking station 6 for the transfer of data between the dispenser 2 and a service provider network server 8, and for the recharging of a battery and/or charge capacitors providing electrical power in the dispenser, in particular its various functions to be described in further detail below.","The docking station includes an electricity mains supply cable 9 and, internally, a voltage transformer for the supply of power at a required low voltage.","In case of mains supply failure or unavailability the docking station may further include a back-up battery, which may be rechargeable or non-rechargable.","The docking station 6 also includes a communications module 10 for communicating with a corresponding communications module 12 on a network interface device 14.The network interface device 14 includes a network interface module 16 for access to a public data communications network 18, such as the Internet.","Both the docking station and the network interface device 14 may be located in a patient's home.","The communications modules 10, 12 preferably communicate via a radio interface protocol, such as the Bluetooth™ radio protocol, thereby allowing the docking station 6 to be located at a selected point in the patient's home, such as a bedroom or bathroom, remote from the network interface device 14.The network interface module 16 may comprise one or more of a PSTN modem, a DSL modem, a cable modem, a wireless radio network modem.","The network interface device may itself take the form of an appropriately configured personal computer workstation, digital television set top box, home communications hub, etc.","FIG.","2 schematically illustrates the electronic components of the medicament dispenser 2, refill 4 and docking station 6 in greater detail.","The dispenser 2 includes a data processor, for example an integrated circuit chip containing a microprocessor 100 and non-volatile memory, in the form of for example ROM 108 and EEPROM 110.Some parts of the memory are reserved for writing to during the manufacture of the dispenser and are, following manufacture, read-only or effectively read-only by virtue of being protected by a secret authentication code, preferably known only by the manufacturer.","These read-only parts include a memory portions storing one or more of a unique identifier for the dispenser, a random or pseudo-random password related thereto, one or more type codes for the one or more different types of medicament the dispenser is intended to be used with, a usage lifetime indicator in the form of an expiry date and/or a prescribed maximum number of usages for the dispenser, manufacturing compliance check data and manufacturer details.","Other parts are writable and are preferably protected, requiring a predetermined authentication step to have been performed before being capable of being written to, or rewritten.","A first set of one or more records are intended to be written initially during dispensing of the dispenser, including one or more of a service provider identifier and/or service provider network address, activation flag, details of an initially prescribed treatment regimen, a version number thereof, and patient name and other patient details.","A second set of one or more records are intended to be written initially to by the docking station 6, including records indicating a variation from the original treatment regimen, a version number thereof, and a deactivation flag.","A third set of one or more records are intended to be written initially to by the dispenser during use, including compliance monitoring data records indicating when medicament has been taken by the patient and what amount thereof.","Some further parts of the memory, containing secret data and/or algorithms, are not externally readable and only for internal use by the microprocessor in performing authentication and encryption procedures.","The dispenser 2 comprises external electrical contacts 20, corresponding to contacts in the docking station, for writing data to and reading data from the dispenser, and for charging the battery and/or capacitors of the dispenser.","The refill 4 includes a data processor, for example an integrated circuit chip containing a microprocessor 116 and non-volatile memory, in the form of for example ROM 118 and EEPROM 120.Some parts of the memory are reserved for writing to during the manufacture of the refill and are, following manufacture, read-only or effectively read-only by virtue of being protected by a secret authentication code, preferably known only by the manufacturer.","These read-only parts include one or more memory portions storing one or more of a unique identifier for the refill, a type code for the stored medicament, a medicament name, an expiry date, an original dosage amount, manufacturing compliance check data and manufacturer details.","Other parts are writable and are preferably protected, requiring a predetermined authentication step to have been performed before being capable of being written to, or rewritten.","A first set of one or more records are intended to be written initially during dispensing of the refill, including one or more of an activation flag, details of a prescribed treatment regimen, a version number thereof, and patient name and other patient details.","A second set of one or more records are intended to be written initially to by the medicament dispenser, including one or more records indicating a current dosage amount remaining.","Some further parts of the memory, containing secret data and/or algorithms, are not externally readable and only for internal use by the microprocessor in performing authentication procedures.","The refill 4 comprises external electrical contacts 22, corresponding to contacts in the dispenser 2, for writing data to and reading data from the refill.","The docking station 6 includes a data processor in the form of for example an integrated circuit chip containing a microprocessor 122 and non-volatile memory, in the form of for example ROM 128 and EEPROM 130.Some parts of the memory are reserved for writing to during the manufacture of the docking station and are, following manufacture, read-only or effectively read-only by virtue of being protected by a secret authentication code, preferably known only by the manufacturer.","These read-only parts include one or more memory portions storing one or more of a unique identifier for the docking station, manufacturing compliance check data and manufacturer details.","Other parts are subsequently writable.","A first set of one or more records are intended to be written to from the dispenser when the dispenser 2 is docked with the docking station, including a medicament name, a current remaining dosage amount, details of a current treatment regimen and patient name and other patient details.","These first records contain data to be displayed on a screen 24.Screen 24 is larger than display 28, and is capable of displaying bitmap images, for example logos, still and video pictures, etc, as well as alphanumeric text, responsive to user interaction via an MMI such as buttons 26.A second set of one or more records are intended to store message data received from service provider network server 8, including one or more records for storing data for messages to be shown on screen 24.A third set of one or more records are for storing control messages to be transmitted to dispenser 2, such as control messages for performing authentication and control messages indicating a variation in treatment regiment, indicating a current dosage amount remaining.","A fourth set of records are for storing data for transmission to network server 8, including compliance monitoring data received from dispenser 2, and self-test health check data, and self-test health status data records, containing data such as peak flow rate data in the case of a respiratory inhaler or blood glucose level data in the case of an insulin dispenser, which is recorded by a sensor 132 in, or associated with, the docking station.","In an alternative, the self-test health check data may be replaced by patient self-assessment data, wherein the patient inputs, via MMI 26, a self-assessed health condition indicator, for example a figure from 1 to 10, indicative of how the patient feels.","The patient may also append the data with text to record additional data relating to their state of health.","The self-test health check data, condition indicator data and other data are automatically date stamped for subsequent analysis by network server 8 and presentation, for example via a patient-specific Web site page, via charts and other assessment tools.","Dispenser display 28, for example an LCD screen, is for displaying information to the patient including the name of the medicament contained in an inserted refill, a current dose remaining, a current time, etc.","Reminder messages may also be displayed thereon to indicate when a dose of medicament is due to be taken, in accordance with the current treatment regimen.","Such a reminder message may accompanied by a timed alarm indication, given for example by an audible (e.g.","tone), kinematic (e.g.","vibration) and/or visual (e.g.","LED) signal generator.","Patients may control display menu and functions by means of a user interface, such as buttons 30.When administering medicine, patient opens outlet 32 and presses a button 30 to initiate the dispensing of a dose, or multiple doses, of medicament.","The amount dispensed is controlled by dispenser 2 in accordance with a current treatment regimen stored in the memory of the dispenser 2 and/or the refill 4, along with the current date/time and/or stage of treatment.","Dispenser 2 further comprises a dispensing actuator 102, for the automated dispensing of a dose, or multiple doses of medicament.","In one embodiment, the medicament is held in refill 4 in powder, liquid and/or gaseous form and a dispensing actuator is adapted to meter out a required dose from refill 4.In another embodiment, refill 4 contains medicament in the form of elements containing discrete amounts of medicament, for example tablets, capsules, powder compartments, etc.","and dispensing actuator is adapted to automatically dispense one or more discrete doses of medicament.","Dispensing may be initiated by the patient, for example by means of MMI 30.Dispenser 2 further comprises dispensing sensor 104, for sensing the act of dispensing and recording, as monitoring data, the amount, time and date when dispensing occurs.","Clock 106 is used for time monitoring and recordal in the dispenser electronic system.","Data interface 22, consisting of two sets of corresponding electrical contacts, is used for the transfer of data between the refill 4 and the dispenser 2.In an alternative embodiment, the interface consists of a contactless radio interface between a passive tag in the refill and an active reader in the dispenser.","Data interface 114, consisting of two sets of corresponding electrical contacts, is used for the transfer of data between the docking station 6 and the dispenser 2.In an alternative embodiment, the interface consists of a contactless radio interface, for example a Bluetooth™ radio interface which is operable when dispenser is placed in the proximity of the docking station 6.Power interface 114 consists of two sets of corresponding electrical contacts for the transfer of electrical power between docking station power transformer 134, supplied by mains power source connection 9, and rechargeable battery 112 in the dispenser 2, which acts as a power source for all electronic components in the dispenser 2 and the refill 4.Transformer 134 acts as a power source for all electronic components in the docking station 6.Docking station 6 may also include a rechargeable back-up battery in case of mains power failure.","Docking station 6 includes a data processing system including a microprocessor 122, non-volatile memory in the form of ROM 128 and EEPROM 130, display 24, man machine interface elements 26, radio interface module 10, including radio antenna 126, a mains power source connection 9, and self-test module 132.Self-test module 132 includes a patient health sensor, such as a blood glucose level sensor, peak flow sensor, etc, and may be detachably connectable to the docking station.","In an alternative embodiment, self-test module 132 is included in dispenser 2, or may be detachably connectable to the dispenser 2.Service provider network server 8 is connected to patient record database 34, which stores long-term patient health records to be used by the health system for the treatment of patients under its care.","These records store historical patient health and treatment data, including prescription data, treatment regimen compliance data, and health check status data.","Service provider network server includes analysis and reporting functions for operating on data contained in the patient health records and for transmitting messages and reports to other healthcare management systems including patient healthcare management systems, physicians' data management systems and pharmacy data management systems.","Manufacturer data management system E includes a manufacturing system server 36, a data writing terminal 38 for writing data to a dispenser 40 during manufacture, and a data writing terminal 40 for writing data to refill 4 during manufacture.","During manufacture, the dispenser memory 108, 110 is written to contain permanent data in the form of one or more of a unique identifier for the dispenser, a random or pseudo-random password, one or more type codes for the one or more different types of medicament the dispenser is intended to be used with, a usage lifetime indicator in the form of an expiry date and/or a maximum number of usages for the dispenser, a geographical region name and/or code, manufacturing compliance check data and manufacturer details.","The dispenser memory is also written to with secret data and/or algorithms, which are not externally readable and only for internal use by the dispenser microprocessor in performing authentication and encryption procedures.","This secret data includes a secret authentication key, which may be of a shared secret type of a private key of a public/private key combination.","The data may also include a public key for one or more service providers, or a trusted third party providing service provider public key details, to authenticate and/or decrypt control messages received from service provider network server 8, which are encrypted and/or digitally signed by the server 8 using its private key and/or the appropriate shared secret key before transmission.","The data may also include a public key for one or more manufacturers, or a trusted third party providing manufacturer public key details, to authenticate and/or decrypt data received from refill 4, which are encrypted and/or digitally signed by the manufacturer or on the refill before transmission.","Following manufacture, the data written to the dispenser memory, including in particular the unique identifier and the corresponding password and/or shared secret key and/or public key, are transmitted in a secure fashion to service provider network server 8 and stored therein awaiting dispensing of the dispenser 2 to occur.","The dispensers are batched and distributed in batches to pharmacies for subsequent dispensing to patients.","During manufacture, the refill memory 118, 120 is written to contain permanent data in the form of one or more of a unique identifier for the refill, a type code for the stored medicament, a medicament name, an expiry date, an original dosage amount, a geographical region name and/or code, manufacturing compliance check data and manufacturer details.","The permanent data also includes a digital signature, generated by manufacturer using a secret authenticating key, of the unique identifier and/or other data, which is added to the refill memory.","The refill memory may also be written to with secret data and/or algorithms, which are not externally readable and only for internal use by the refill microprocessor in performing authentication procedures.","This secret data may include a secret authentication key, which may be of a shared secret type of a private key of a public/private key combination.","The data may also, or alternatively, include a public key for the service provider, to authenticate and/or decrypt control messages received from service provider network server 8, which are encrypted and/or digitally signed by the server 8 using its private key and/or the appropriate shared secret key before transmission.","Following manufacture, the data written to the refill memory, including in particular the unique identifier and the corresponding digital signature, shared secret key and/or public key, are transmitted in a secure fashion to service provider network server 8 and stored therein awaiting dispensing of the refill 4 to occur.","The refills are batched and distributed in batches to pharmacies for subsequent dispensing to patients.","Patient system A also contains a smart card 42 for identifying the patient and storing patient healthcare details, which preferably are replicated in patient record database 34, including general health data, prescription data, allergy data, etc.","Smart card 42 includes electrical contacts 43 whereby a smart card reader/writer accesses data records held on the smart card.","Physician data management system C includes a workstation personal computer 44, a smart card reader/writer 46 and a dispenser reader/writer 48.The dispenser reader/writer 48 may be a docking station similar to the patient docking station 6.On visiting a physician, a patient preferably carries their smart card 42.The smart card is inserted in reader/writer 46 to allow the physician to access patient data held thereon and/or patient data held in patient record database 34.The data is accessed via workstation 44.Each physician has a personal username and password providing the physician with authority to access such data, which details are verified by network server 8 before such access is granted.","Following consultation, the physician may issue a digital prescription, which is written to smart card 42 and/or patient record database 34.Pharmacy data management system D includes a server 50, a workstation personal computer 52, a dispenser reader/writer 54, a refill reader/writer 56 and a smart card reader/writer 58.The dispenser reader/writer 54 may be a docking station similar to the patient docking station 6.When a patient visits the pharmacy to obtain supplies, the patient carries their smart card 42, which is inserted in smart card reader 58 in order to identify the patient and the corresponding prescription.","The patient data is accessed via workstation 52.Each pharmacist has a personal username and password providing the pharmacist with authority to access such data, which details are verified by network server 8 before such access is granted.","The patient identity is transmitted to network server 8 in order to access and/or the prescription data.","The patient's data record indicates whether the patient currently holds a dispenser appropriate for the prescribed medicament.","If not, as in the case of a newly diagnosed patient, the network server instructs pharmacist system to dispense a new dispenser 2.In order to dispense a new dispenser 2, the pharmacist retrieves the appropriate dispenser from the pharmacy stores and inserts the dispenser into dispenser reader/writer 54.The reader/writer 54 reads the dispenser's unique identity and transmits same to network server 8.FIG.","3 shows a look-up table held database in 34, associating each unique dispenser ID, and the associated data fields, corresponding to the original data held in the dispenser memory, provided by manufacturer (illustrated as Field #1 etc.)","with a patient ID following dispensing thereof.","The reader/writer is then used to write data received from network server 8, containing one or more of a service provider identifier and/or service provider network address, activation message, details of an initially prescribed treatment regimen, a version number thereof, patient ID and name and other patient details, and a geographical region code, to the dispenser memory.","The written data is preferably encrypted and/or signed by the network server 8, thereby indicating to the dispenser that a pharmacist having the necessary authority is performing the dispensing operation.","Once activated, the dispenser 2 is ready for use with an appropriate refill 4.In order to dispense a new refill 4, the pharmacist retrieves the appropriate refill from the pharmacy stores and inserts the dispenser into dispenser reader/writer 54.The reader/writer 54 reads the refill's unique identity and transmits same to network server 8.FIG.","2 shows a look-up table held database in 34, associating each unique refill ID, and the associated data fields, corresponding to those originally held in the refill memory, provided by manufacturer (illustrated as Field #1 etc.)","with a patient ID following dispensing thereof.","The reader/writer is then used to write data provided by network server 8, containing one or more of an activation message, details of a prescribed treatment regimen, a version number thereof, and patient ID, name and other patient details, and a geographical region code, to the refill memory.","The written data is preferably encrypted and/or signed by the network server 8, thereby indicating to the refill that a pharmacist having the necessary authority is performing the dispensing operation.","Once activated, the refill 4 is ready for use with an appropriate dispenser 2.In one embodiment, the pharmacy procedures include reading the data stored on the dispenser and/or refill prior to dispensing, and modifying the dispensing operation accordingly, either in the pharmacy system alone or in combination with the network server system.","For example, the pharmacy may be prevented from dispensing a device having a geographical region name and/or code which does not correspond for refills and dispensers, or if the location of the pharmacy does not correspond therewith.","The look-up tables shown in FIGS.","3 and 4 are used for the correct identification of a patient by means of the corresponding dispenser and/or refill ID's.","When data is transferred from a dispenser 2 and/or a refill 4 to network server 8, the data includes the device ID and/or the refill ID.","Thereby, the patient is identified without requiring the patient to input username and/or password details each time data is sent to the network server 8.Furthermore, the associated authentication procedures may be performed by means of the devices themselves, rather than the patient having to identify and carry out authentication each time sensitive data is transferred.","The corresponding shared authentication data, such as shared secret keys, held on both the devices and in the device-related parameter fields illustrated in FIGS.","3 and 4, may be uniquely correlated by means of the look-up tables to the identity of the patient to whom the device was prescribed.","In an alternative embodiment, the unique patient ID is written at the pharmacist onto the dispenser and/or refill during dispensing, and the patient ID is sent with any data sent from dispenser to network server 8.However, in this case a device ID, which may be unique amongst all devices, or unique only in combination with the patient ID, is preferably also sent to allow the identification of the device currently being used by the patient at the network server 8.In any case, the data transmitted to network server 8 contains a unique identity, preferably device ID, whereby the corresponding patient record may be accessed.","FIG.","5 is a flow diagram illustrating procedures carried out in the dispenser 2 on insertion of a refill 4.On insertion of the pharmacy-dispensed refill 4 into the pharmacy-dispensed dispenser 2, the dispenser reads all available data held in refill memory, step 200.The dispenser then checks that no conflicts exist between the data read and that stored in its memory, steps 202 and 204, including ensuring that the refill has a matching patient ID, acceptable medicament type code, acceptable geographical region code, etc.","Furthermore, dispenser 2 checks that the any digital signature accompanying the parameter data held in refill is correct.","This check is carried out, in one embodiment, by reading the appropriate signature and decrypting it with the public key of the network server 8 to ensure that it corresponds with the original data, for example the unique identity of the refill 4, to which the signature is attached.","In other embodiments, other cryptographic methods are used to check the authenticity of the refill by means of public/private key or shared secret key encryption/decryption methods.","If a conflict is found, a conflict indicator signal is generated.","If no conflict is found, the dispenser checks if there are any updates to be received from the refill.","For example, the refill may contain a more recent treatment regimen, which may be detected by means of a later version number.","If an update is detected, it is first authenticated, by checking the accompanying digital signature, before updating the appropriate parameters in its memory.","If the update authentication fails, a conflict indicator signal is generated.","Otherwise, following any update, the dispenser 2 begins a normal treatment regimen in accordance with data held on the dispenser and/or the refill 4.If a conflict indicator signal is generated, the dispenser display 28 is activated to show an appropriate error message, for example “Refill not authorised: see pharmacist”, step 206.The dispenser 2 also inhibits any reminder programme which would otherwise have been followed, step 208.On generation of the conflict indicator signal, the dispenser may, in one embodiment, write a deactivation flag in the dispenser or refill memory to prevent the refill and/or the dispenser being used, at least until the conflict is resolved by means of an update message received from server 8.Alternatively, the dispenser may still be used in manual override mode to dispense medicament from the refill, even though the data management functions are not made available.","Further alternatively, the dispenser may continue to operate normally but store the conflict data to transmit same to network server 8 when the dispenser is next docked.","The action taken by dispenser may depend upon the nature of the conflict found.","For example, if the refill contains the wrong type of medicine, the refill may be fully deactivated.","On the other hand, if the refill was dispensed to a different patient, but contains the correct type of medicament, the dispenser may be used as normal, with the conflict data being sent to network server 8 to be written to the patient record in order that a healthcare professional, such as the physician or pharmacist, may be made aware of the conflict when the patient record is next accessed.","The dispenser then senses whether a docking connection is made to docking port 6, for example by means of a data read signal being received from docking port 6, at which point an message indicating the detected conflict is sent, via docking station 6, to network server 8, steps 210 and 212.Network server 8 may used the conflict data for monitoring purposes, and inform the appropriate healthcare professional as and when required of the detected conflict.","For example, when patient next visits a pharmacist, the pharmacist will be able to access, via the patient record, the conflict data and indicated the nature of the problem to the patient.","If the conflict is an internal conflict caused by an error in the network server system 8, the conflict may be resolved and updates may be sent to all other affected dispensers to avoid the conflict occurring unnecessarily elsewhere.","Preferably, the dispenser may still be used in manual override mode to dispense medicament from the refill, even thought the data management functions are not made available.","In any case, when a conflict is found, conflict data indicating the cause of the conflict, e.g.","both the data derived from the refill and the corresponding, incompatible, data held on dispenser, is transmitted to network server 8.Network server 8 analyses the nature of the conflict and can take remedial action, such as writing corresponding data to patient record for perusal by a healthcare professional having access to the system, transmitting informational messages to the patient via the patient docking station, and/or transmitting update messages to the dispenser via the docking stations to resolve the conflict by updating one or more fields of the dispenser memory as appropriate.","If no conflict is found, the unique identity of the refill and/or other details held on the refill are written to a record in the dispenser memory for transfer, via docking station 6, to the network server 8 when the dispenser is next docked.","Such data, derived from the refill, is then stored in the appropriate patient record by the network server 8, to provide a medication history, including details of the refill container used, for the patient.","Furthermore, in an alternative embodiment, any conflict checks are carried out by network server.","In this embodiment, the current refill ID and device ID are preferably transmitted to network server 8, and the corresponding parameter data, held in the lookup tables illustrated in FIGS.","3 and 4, is analysed by network server 8 to determine whether a conflict exists.","Alternatively, one or more of the other parameter data items held in the dispenser memory and/or the refill memory may be transmitted to network server 8.If a conflict is found, network server 8 analyses the nature of the conflict and can take remedial action, such as writing corresponding data to patient record for perusal by a healthcare professional having access to the system, transmitting informational messages to the patient via the patient docking station, and/or transmitting update messages to the dispenser via the docking stations to alter the functioning of the dispenser, to disable the dispenser until a new refill has been inserted, and/or to resolve the conflict by updating one or more of the previously incompatible fields of the dispenser memory as appropriate.","FIG.","6 illustrates a treatment procedure followed by dispenser 2 when loaded with a correct refill 4.The dispenser holds treatment regimen data from which it is determined when a medicament dose is due to be taken by the patient, step 300.A period is defined when the dose is defined to be correctly taken, by means of a start date/time and an end/date time, which may be defined relative to a last taken medication or in accordance with an absolute timing policy.","If the patient activates the dispenser prior to the start date/time, step 302, the dispenser acts in accordance with an early dispensing procedure, step 304, which is different to the normal dispensing procedure.","This may be to at least initially inhibit dispensing and display a message to the patient indicating that it is too early to take another dose.","An option may at this stage be given to allow the patient to confirm that a dose is required, in response to which the dispenser conducts dispensing as required.","Alternatively, dispenser may initially dispense the dose at the requested time, and update the subsequent reminder schedule to cancel the upcoming reminder and bring forward the subsequent reminders.","When a dosing reminder is due, the dispenser 2 emits a dosing reminder signal, step 306.The signal may be an audio, visual and/or kinematic signal, and may vary over time depending on whether dosing is carried out by the patient, for example to increase in intensity over time.","When patient actuates the dispenser to dispense a dose of medicament, step 310, the dispenser determines whether the dispensing operation is later than the predefined end date/time for the normal dosing period, step 310.If so, a late dispensing procedure, different to a normal dispensing procedure, is carried out, step 318.This may consist of indicating to the patient that the dose is being taken late and adjusting the subsequent reminder schedule accordingly, to delay subsequent reminders in accordance with the delay.","If within the normal dosing period, the normal dosage data is accessed, step 312, and the appropriate dosage is dispensed, step 314.Following all dispensing procedures, although only shown in relation to normal dispensing procedure, the monitoring data held in dispenser, subsequently to be sent to network server 8, is updated to store details of the latest dispensing event, including date/time, amount dispensed, and any other pertinent data such as patient health data (e.g.","peak flow rate) detected during the dispensing operation.","FIG.","7 illustrates a procedure carried out by docking station 6 when a dispenser is docked therein.","Note that in this preferred embodiment, the docking station 6 may be used with multiple different dispensers, including multiple dispensers owned by the same patient and/or multiple dispensers owned by different patients.","On receiving a dispenser 2, the docking station 6 first reads the patient name and medicament name, along with other pertinent data such as number of doses remaining, etc., and displays these details to the patient, steps 400, 402, via display 24.The patient may scroll through the relevant data held on dispenser by means of an MMI interaction.","Next, the docking station reads other stored data from the dispenser, including the dispenser identity, the refill identity, any conflict data resulting from a conflict check conducted by dispenser 2, and dispensing monitoring data stored when the patient has dispensed medication from the dispenser 2.This stored data may be provided by the dispenser in encrypted form, for example encrypted using its stored secret key, for transmission by the docking station in encrypted form, step 406, and decryption by the network server 8 with reference to the corresponding decryption key held in the details held for the identified dispenser.","Furthermore, authentication data, in the form of a digital signature produced by the dispenser 2 and/or the refill 4, may also be transmitted for checking by network server.","The docking station may form a suitable data connection, for example a TCP/IP socket connection, to the network server 8.The connection between the docking station 6 and the network server 8 may in any case be arranged to be secure, for example via secure socket layer (SSL) interactions.","The data sent by docking station is analysed at the network server 8 and one or more response messages are generated and sent to be received by docking station, step 408.These messages may be structured as extensible markup language (XML) documents.","The responses preferably comprise data to be displayed to user via the display screen 24 of the docking station, and update data to be passed by docking station.","Docking station 6 parses the response data, step 410, displays the display content, step 412, and transmits any update messages to the dispenser 2, step 414.The update messages are preferably encrypted by network server 8 using the corresponding encryption key, for example the dispenser's shared secret key or the public key of the dispenser, and/or contain a further authentication element, such as a digital signature authenticating the data as having been generated by, or with the authority of network server 8.FIG.","8 illustrates steps taken by dispenser 8 on receipt of an update message via docking station 6, step 500.Initially, the dispenser 2 conducts decryption of the data using a secret key stored in the dispenser memory, step 502.Next, the dispenser checks the accompanying digital signature, step 502.If either of these steps fails, the update message cannot be authenticated, step 506 by dispenser 2 and an error message is displayed, step 510.An error message may also be sent to network server 8, to allow a further, corrected, update to be sent.","If the update is authenticated by the dispenser, step 506, the corresponding data fields are updated in the dispenser memory and/or the refill memory, step 508.It is to be noted that any one or more of the writable fields of the dispenser and/or the refill, such as any of those previously mentioned, may be updated in this way.","If the dispenser 2 and/or the refill 4 fails to authenticate to network server 8, the network server can send an appropriate message to the docking station, such as “Authentication failed: please visit pharmacy”.","In a subsequent physician visit, if the patient has brought their dispenser 2, and the dispenser is inserted into reader/writer 48 in order for the most recent compliance and health status data to be downloaded to network server 8.The dispenser's unique identity is transmitted to network server 8, along with authentication data in the form of the corresponding password and/or, if stronger authentication is required, an encrypted response to a challenge message sent by the server 8, which response is verified by server 8 in order to validate the dispenser identity.","The corresponding patient identity is looked up by network server in order to access the correct patient record.","Not only does physician have access to patient data, but also the analysis and reporting functions provided by network server 8 following the period of patient compliance and health monitoring provided by dispenser 2.Following consultation with the network server functions and the patient, the physician may vary the treatment regimen, by input to workstation 44 which results in an update procedure whereby network server 8 transmits data to be written to the dispenser by reader/writer 48.The transmitted data is encrypted and/or digitally signed by the network server, using the appropriate secret and/or private key for the dispenser 2.On receipt, the dispenser decrypts and/or authenticates the control data and, if authentication is successful, updates its records accordingly.","The physician may also prescribe more or a new medicament, in which case a digital prescription is written to smart card 42 and/or to patient record database 46.If on docking with docking station 6, either the docking station or the network server 8 establishes that the patient is due for another prescription, a message may be displayed to the patient inquiring whether a further prescription should be ordered.","If the patient replies in the affirmative, a represcription order placed in the patient record, or is transmitted to a selected pharmacist, and the patient may pick up the appropriate refill by presenting at the pharmacist with their patient smart card 42.The network server-based system also monitors the lifetimes of dispensers and/or refills.","Namely, associated with each of the dispensers and refills, by means of parameter data held against the device identity in the look-up tables illustrated in FIG.","4, is an expiry date for the device, and when the device is due to expire, a suitable message may be sent to docking station instructing the patient to order the prescription of a new device.","On expiry an update message may be sent to the dispenser via the docking station to disable dispensing.","The lifetime of the dispenser may be based solely on an expiry date, a usage amount (e.g.","maximum number of allowed uses), or both.","In any case, such monitoring and messaging is useful in order to inhibit the at least inconvenient failure of any of the devices used by the patient.","In the above-described embodiments, the patient is identified primarily by means of the dispenser and/or the refill which they have been dispensed by a registered pharmacist.","Additional identity checks may be carried out by the system.","For example, the dispenser 2 or the docking station 6 may include a Biometric sensor such as a fingerprint sensor for positively identifying the patient before any actions based upon the patient identity are carried out.","In different embodiments of the invention, the dispenser is typically shaped to define a cavity within which the refill is receivable.","The dispenser and/or refill may be further shaped with grooves, indentations or other shaping or surface details to define a slidingly lockable/releasable relationship between the dispenser and the refill.","Accordingly, the dispenser may have a lockable cover which is releasable to expose the cavity within which the refill is held.","FIGS.","9a and 9b show a metered dose inhaler comprising a dispenser in the form of a tubular actuator housing 601 shaped for receipt of an aerosol refill 602.The actuator housing is open at one end and is closed at the other.","An outlet 603 leads laterally from the closed end of the housing 601.In the embodiment illustrated, the outlet 603 is in the form of a mouthpiece intended for insertion into the mouth of the patient but it may, if desired, be designed as a nozzle for insertion into the patient's nostril.","The aerosol refill 2 has an outlet valve 604 at one end.","This valve acts as a release means for release of a measured dose from the aerosol refill.","The release means is actuable by inward movement of the valve 4 relative to the aerosol refill 602.The metered dose inhaler of FIGS.","9a and 9b includes an electronic data management system 610, including all electronic components and electrical connections, and is adapted for docking with a corresponding docking station, as described above in detail, comprised within an extended part 606 of the housing 6.A visual display (not visible) allows for display of information from the electronic data management system 610 to the patient.","The electronic data management system connects to a sensor (not visible) for sensing the breathing pattern of the patient and an actuator (not visible) for actuating the release of aerosol from the refill 2.A data module in the form of a chip 620 is mounted on the side of the aerosol refill 602.Corresponding read/write contacts 630 for the chip 620 are moulded into the internal surface of the dispenser housing 601.In an alternative embodiment the data module 620 comprises a radiofrequency identification tag and the read/write module 630 is capable of reading data therefrom and writing data thereto by the use of interrogating radiofrequency energy.","In another embodiment the data module 620 comprises a magnetic label and the read/write module 30 is capable of reading data therefrom and writing data thereto by the use of interrogating magnetic field energy.","FIG.","10 shows a schematic representation of a breath-operable medicament dispensing system, which is an embodiment of the system shown in FIGS.","9a and 9b.","The system comprises a metered dose inhaler similar to that shown in FIGS.","9a and 9b, comprising tubular housing 710 having a dispensing outlet 712 in the form of a mouthpiece.","Within the housing 710 sits aerosol refill 720 which has a valve dispensing mechanism 722 in the form of a slide valve.","Valve stem 724 is supported by valve support 714.Outlet passage 716 is provided in the support 714 to enable passage of dispensed dose to the dispensing outlet 712.It may be seen that the upper part of the aerosol refill 720 abuts refill seat 730.The refill seat 730 comprises an insulating portion 732 which directly contacts the aerosol refill 720 and an upper conducting portion 734 (e.g.","comprised of aluminium).","It may also be seen that the valve support 774 connects with conducting valve seat 740.Plural shape memory alloy wires 750a, 750b connect the conducting portion 734 of the refill seat 730 to the conducting valve seat 740.The plural wires 750a, 750b comprise a nickel-titanium alloy which contracts in response to electrical current flow therethrough.","It may thus, be appreciated that on passage of electrical current through the plural wires 750a, 750b the refill seat 730 and valve seat 740 will be drawn towards each other as the wires 750a, 750b contract.","Actuation of the valve dispensing mechanism 722 and dispensing of medicament dose will thereby result.","Control of electrical current flow to the refill seat 730, valve seat 740 and wires 750a, 750b is achievable using the illustrated circuitry.","Refill seat 730 and valve seat 740 connect to actuation circuit 760 which includes a high current power supply 762 (e.g.","a voltaic cell or battery of voltaic cells) and a switch 764 in the form of a solid state relay.","The solid state relay 764 itself connects with control circuitry including a micro-controller 770 having an independent power supply 772.The micro-controller 770 itself connects with pressure transducer 780 which has an input in the form of a pressure tube 782 located within the dispensing outlet 772 of the inhaler housing 770.It may be appreciated that current flow to the refill seat 730, valve seat 740 and wires 750a, 750b, and hence actuation of the valve dispensing mechanism may be achievable as follows.","The patient inhales through the mouthpiece 772 resulting in a change in pressure within the housing 770 and pressure tube 782.The change in pressure is detected by the pressure transducer 780 which sends a signal to the micro-controller 770.The micro-controller 770, in turn sends a switching signal to the solid state relay 764 which results in closing of the actuation circuit and electrical current flow therethrough.","The resulting contraction of the shape memory alloy wires 750a, 750b causes actuation of the valve dispensing mechanism 722 and hence, dispensing of medicament to the inhaling patient.","It may also be seen in FIG.","10 that the micro-controller 770 is connected to a display 774 for display of information to the patient and also with a computer interface 776 for exchange of data therewith.","All circuitry and components thereof including the power supplies 762, 772 and display 774 may be arranged to be present on the housing 770 such that the system is in the form of a discrete, hand-held device.","FIG.","11 shows a medicament dispenser 800 containing a refill cartridge 802 in accord with a further embodiment of the present invention.","The medicament refill cartridge 802 comprises a flexible strip 804 defining a plurality of pockets each of which contains a discrete dose of medicament.","The doses may be in the form of tablets, capsules, doses of powder which can be inhaled, etc.","The strip 804 comprises a base sheet in which blisters are formed to define the pockets and a lid sheet which is hermetically sealed to the base sheet except in the region of the blisters in such a manner that the lid sheet and the base sheet can be peeled apart.","The lid and base sheets are each preferably formed of a plastics/aluminium laminate and are preferably adhered to one another by heat sealing.","FIG.","11 shows the internal mechanism of medicament dispenser 800 in the situation where the majority of the pockets of the refill strip 804 are still filled with medicament.","The internal mechanism comprises an index wheel 806 and a lid-winding spool 808 for winding the used portion of the lid sheet.","The index wheel 806 has a plurality of recesses extending parallel with the axis of the wheel.","The recesses are spaced at a pitch which is equal to the distance between the centre lines of adjacent pockets in the refill strip 804.The refill 802 includes an internal area for the medicament strip 802 to be coiled in prior to use of the doses contained inside it.","The dispenser comprises an area where the used base and lid of the medicament carrier strip 804 is collected.","After complete usage, the used base may be rewound back into refill automatically prior to removal of the refill.","The dispensing system 800, 802, includes an electronic data management system 810, including all electronic components and electrical connections, as described above in detail, and is adapted for docking with a corresponding docking station as described in detail above.","The refill 802 includes a data module 812 in the form of a chip, as described above, which interfaces with the data management system 810 when the refill 802 is inserted in the dispenser 800, via a hinged insertion door 814, which is secured in place by means of a catch 816.On insertion of the refill, the lid sheet is wrapped around spool 808 and the base sheet, containing the pockets of medicament, is inserted in transport passage 818 of the dispenser.","Spool 808 and/or index wheel 806 are revolved, by means of a motor controlled by data management system 810 a predetermined amount for each dispensing operation.","As the dispensing operation occurs, a new dose of medicament is presented at outlet 820 for consumption by the patient.","Medicament dispensers of the present invention are in one aspect suitable for dispensing medicament for the treatment of respiratory disorders such as disorders of the lungs and bronchial tracts including asthma and chronic obstructive pulmonary disorder (COPD).","Appropriate medicaments may thus be selected from, for example, analgesics, e.g., codeine, dihydromorphine, ergotamine, fentanyl or morphine; anginal preparations, e.g., diltiazem; antiallergics, e.g., cromoglycate (e.g.","s the sodium salt), ketotifen or nedocromil (e.g.","as the sodium salt); antiinfectives e.g., cephalosporins, penicillins, streptomycin, sulphonamides, tetracyclines and pentamidine; antihistamines, e.g., methapyrilene; anti-inflammatories, e.g., beclomethasone (e.g.","as the dipropionate ester), fluticasone (e.g.","as the propionate ester), flunisolide, budesonide, rofleponide, mometasone e.g.","as the furoate ester), ciclesonide, triamcinolone (e.g.","as the acetonide) or 6α,9α-difluoro-11β-hydroxy-16α-methyl-3-oxo-17α-propionyloxy-androsta-1,4-diene-17β-carbothioic acid S-(2-oxo-tetrahydro-furan-3-yl) ester; antitussives, e.g., noscapine; bronchodilators, e.g., albuterol (e.g.","as free base or sulphate), salmeterol (e.g.","as xinafoate), ephedrine, adrenaline, fenoterol (e.g.","as hydrobromide), formoterol (e.g.","as fumarate), isoprenaline, metaproterenol, phenylephrine, phenylpropanolamine, pirbuterol (e.g.","as acetate), reproterol (e.g.","as hydrochloride), rimiterol, terbutaline (e.g.","as sulphate), isoetharine, tulobuterol or 4-hydroxy-7-[2-[[2-[[3-(2-phenylethoxy)propyl]sulfonyl]ethyl]amino]-ethyl-2(3H)-benzothiazolone; adenosine 2a agonists, e.g.","2R,3R,4S,5R)-2-[6-Amino-2-(1S-hydroxymethyl-2-phenyl-ethylamino)-purin-9-yl]-5-(2-ethyl-2H-tetrazol-5-yl)-tetrahydro-furan-3,4-diol (e.g.","as maleate); α4 integrin inhibitors e.g.","(2S)-3-[4-({[4-(aminocarbonyl)-1-piperidinyl]carbonyl}oxy)-phenyl]-2-[((2S)-4-methyl-2-{[2-(2-methylphenoxy)acetyl]amino}pentanoyl)-amino]propanoic acid (e.g.","as free acid or potassium salt), diuretics, e.g., amiloride; anticholinergics, e.g., ipratropium (e.g.","as bromide), tiotropium, atropine or oxitropium; hormones, e.g., cortisone, hydrocortisone or prednisolone; xanthines, e.g., aminophylline, choline theophyllinate, lysine theophyllinate or theophylline; therapeutic proteins and peptides, e.g., insulin or glucagon; vaccines, diagnostics, and gene therapies.","It will be clear to a person skilled in the art that, where appropriate, the medicaments may be used in the form of salts, (e.g., as alkali metal or amine salts or as acid addition salts) or as esters (e.g., lower alkyl esters) or as solvates (e.g., hydrates) to optimise the activity and/or stability of the medicament.","Preferred medicaments are selected from albuterol, salmeterol, fluticasone propionate and beclomethasone dipropionate and salts or solvates thereof, e.g., the sulphate of albuterol and the xinafoate of salmeterol.","Medicaments can also be delivered in combinations.","Preferred formulations containing combinations of active ingredients contain salbutamol (e.g., as the free base or the sulphate salt) or salmeterol (e.g., as the xinafoate salt) or formoterol (e.g.","as the fumarate salt) in combination with an anti-inflammatory steroid such as a beclomethasone ester (e.g., the dipropionate) or a fluticasone ester (e.g., the propionate) or budesonide.","A particularly preferred combination is a combination of fluticasone propionate and salmeterol, or a salt thereof particularly the xinafoate salt).","A further combination of particular interest is budesonide and formoterol (e.g.","as the fumarate salt).","The medicament dispenser of the present invention in other embodiments comprises a syringe for the delivery of injectable medicament to a patient.","Traditional syringes rely on puncturing of the patient's skin by a hollow needle through which the injectable medicament (in solution or suspension form) is delivered to the muscle or tissue of the patient.","Recently developed needleless systems for the delivery of injectables employ high velocity injection of particle formulated drugs or vaccine through the skin and into any physically accessible tissue.","Other needleless systems employ similar high velocity injection of drug or vaccine coated on to a suitable carrier particle.","It will be understood that the present disclosure is for the purpose of illustration only and the invention extends to modifications, variations and improvements thereto.","In alternative embodiments, there is no docking station situated in the patient home or other environment.","However a docking station 6 as described may be provided in a health care provider location, such as in patient clinic, physician's surgery or pharmacist.","In this, and other alternative embodiments, the dispenser contains a sealed long-life battery.","In the above embodiments, the service provider authentication and decryption procedures for updates messages are carried out by the dispenser 2.In an alternative embodiment, the authentication and/or decryption is performed by the docking station 6.In the above embodiments, the refill is filled with medicament at the site of manufacture.","In an alternative embodiment, the refill may be filled in pharmacy, and its memory written to with data identifying its contents and expiry date therefore.","In the above embodiments, authentication is performed by means of encryption/decryption and by means of digital signature checking.","Other known methods of authentication may also, or alternatively be used.","Herein the term digital signature is intended to include digital certificates, which also provide means of authentication.","In the above embodiments, the docking station must first establish a connection with the network server in order for update and display messages to be sent to the patient.","Other means of delivery of messages, in particular push messages sent by the server at a time determined by the server, may be sent in other formats, for example by means of cellular telephony SMS messages, and to other devices, such as cellular radio devices.","In this description, and in the following claims, the devices involved in the present invention include a dispenser and a refill.","It is to be noted that the dispenser is not limited to a device comprising a medicament outlet; the outlet may be provided on the refill.","The dispenser is a device containing elements for the control of a dispensing operation in which medicament, originally contained in the refill, is dispensed."]],"string":"[\n [\n \"Medicament dispenser\",\n \"Apparatus for dispensing medication to a patient, said apparatus comprising: (a) an electronic medicament dispenser (2) with one or more data-transfer elements; (b) a docking station (6) with one or more corresponding data-transfer elements; and (c) a wireless communications module (10) for connecting said docking station to a public communications network, whereby data may be transmitted remotely between said medicament dispenser and a remote processing node via said public communications network.\"\n ],\n [\n \"1.Apparatus for dispensing medication to a patient, said apparatus comprising: (a) an electronic medicament dispenser with one or more data-transfer elements; (b) a docking station with one or more corresponding data-transfer elements; and (c) a wireless communications module for connecting said docking station to a public communications network, whereby data may be transmitted remotely between said medicament dispenser and a remote processing node via said public communications network.\",\n \"2.Apparatus according to claim 1, wherein said wireless communications module forms an integral part of said docking station.\",\n \"3.Apparatus according to claim 1, wherein said data-transfer elements comprise electrical contacts.\",\n \"4.Apparatus according to claim 1, wherein said docking station comprises a display for displaying data relating to said medicament dispenser.\",\n \"5.Apparatus according to claim 1, wherein said medicament dispenser comprises a display, said medicament dispenser display being smaller in area than said docking station display.\",\n \"6.Apparatus according to claim 1, wherein said medicament dispenser comprises means for authenticating data received from said docking station 7.Apparatus according to claim 1, wherein said docking station comprises means for initiating communications sessions with a network entity in response to docking of said dispenser therewith.\",\n \"8.Apparatus according to claim 1, wherein said dispenser comprises a removable refill container.\",\n \"9.Apparatus according to claim 8, wherein said docking station is arranged to transmit data relating to any conflict between said dispenser and said refill to a network entity via a communications link.\",\n \"10.Apparatus according to claim 1, wherein said docking station is arranged to form a link via said wireless communications module with one or more of an appropriately configured personal computer workstation, a digital television set top box, a digital television and/or a home communications hub.\",\n \"11.A method of controlling the dispensing medication to a patient, the method comprising: (a) providing an electronic medicament dispenser with one or more data-transfer elements; (b) providing a docking station with one or more corresponding data-transfer elements; and (c) providing a wireless communications module for connecting said docking station to a public communications network, (d) transmitting data remotely between said medicament dispenser and a remote processing node via said public communications network, using said docking station and said wireless communications module.\",\n \"12.A method according to claim 11, comprising transmitting data from said medicament dispenser to said remote processing node.\",\n \"13.A method according to claim 11, comprising transmitting data from said remote processing node to said medicament dispenser.\"\n ],\n [\n \"The present invention relates to medicament dispensers for use in the dispensing of medicament, to medicament containers for use in same, and to systems and methods for the dispensing of medicament.\",\n \"Lack of patient compliance with medical treatment regimens is known to be a problem.\",\n \"According to the United States Food and Drug Administration (FDA), between 30 and 50 percent of patients fail to use medicines as prescribed, see “FDA Proposes Program to Give Patients Better Medication Information”.\",\n \"The main problems are known to be taking improper dose, failing to take doses on time and ceasing treatment prematurely.\",\n \"If this problem could be addressed, the quality of care offered to patients could be improved considerably.\",\n \"Medical dispensers are well known for the dispensing of various kinds of medicament.\",\n \"In their most simple form, they may comprise a container of tablets with a removable lid or blister packs of tablets.\",\n \"Inhalation devices, such as metered dose inhalers (MDI) and dry powder inhalers are known for the delivery of medicament for the treatment of respiratory disorders.\",\n \"Syringes, including needleless syringes are also known for the delivery of injectable medicament to a patient.\",\n \"Reloadable medicament dispensers are known.\",\n \"These typically comprise a housing defining a cavity and a medicament container, referred to herein as a refill, which is reversibly receivable thereby.\",\n \"The housing and the medicament container may be sold separately or as a kit of parts.\",\n \"Currently, medicament dispensers are primarily manually operable.\",\n \"A patient actuates the dispenser in order to receive a dose of the medicament.\",\n \"If a variation in the basic dose, e.g.\",\n \"more than one dose, is required, the patient typically manually administers two doses.\",\n \"It would be desirable to provide a way in which the patient is relieved of sole responsibility for the correct administration of doses of medicament, and more preferably the variation of such doses, within a treatment regimen.\",\n \"Further desirable features relating to medicament dispensers include ease of use, portability and ease of manufacture.\",\n \"PCT patent application no.\",\n \"WO92/17231 describes a metered dose inhaler having a microelectronic assembly thereon.\",\n \"The medicament container includes a set of electrically conducting strips which represent information about the medicament container in digital form.\",\n \"The housing of the device includes electrical contact fingers which are contactable with the strips to enable reading of the information to a microelectronic memory on the housing.\",\n \"Read/write communication would not be possible between the fingers and the reader and the significant advantages of the present invention would therefore not be achievable with this inhaler.\",\n \"Furthermore, contact between the strips on the container and the electrical contact fingers is required which requires physical tailoring of the container to the housing, thereby limiting product design options.\",\n \"This document also describes an embodiment in which an active (i.e.\",\n \"powered) microelectronic element is attached to the container.\",\n \"PCT Patent Application No.\",\n \"WO 00/25720 describes a medication container that organises several vials or cassettes of different types of medication by securing the vials to a unitary lid.\",\n \"A machine readable memory strip is affixed to each vial.\",\n \"Each memory contains prescription information and medication implementation pertaining to the medication in the vial.\",\n \"The unitary lid is equipped with sensors that read each memory strip and transmit the information to a processor in the lid.\",\n \"The processor determines when each medication is to be taken and signals the patient to take the appropriate medication from the appropriate vial at the appropriate time.\",\n \"The automated lid also contains a receiver for obtaining updated medication dosing information based on current laboratory tests or physical observations of the physician regarding the patient.\",\n \"It is an object of the present invention to provide improvements in relation to systems in which electronic medicament dispensers are used in a course of treatment.\",\n \"In accordance with one aspect of the present invention there is provided apparatus for dispensing medication to a patient, said apparatus comprising: (a) an electronic medicament dispenser with one or more data-transfer elements; (b) a docking station with one or more corresponding data-transfer elements; and (c) a wireless communications module for connecting said docking station to a public communications network, whereby data may be transmitted remotely between said medicament dispenser and a remote processing node via said public communications network.\",\n \"By providing an electronic medicament dispenser with one or more data-transfer elements, and a docking station with one or more corresponding data-transfer elements, functionality which is not essential to the short-term functioning of the medicament dispenser may be placed in the docking station.\",\n \"By providing a separate docking station, features which are not essential to the short-term functioning of a dispenser may be instead implemented in the docking station, thereby increasing the versatility of the system.\",\n \"These features may include a data transfer feature.\",\n \"The provision of a wireless communication module for connecting the docking station to a public communications network allows the functioning of the docking station to be augmented by resources provided in a remote processing node, whilst allowing the location of the docking station within the user's environment to be selected by the user.\",\n \"In contrast, requiring the user to locate a docking station at a public data communications access point, such as a telephone access point, would in many cases prevent the frequent and correct usage of the device which is desired in a system in accordance with the present invention.\",\n \"Further features and advantages of the present invention will become apparent from the following description of preferred embodiments of the invention.\",\n \"Embodiments arranged in accordance with the invention will now be described, by way of example only, with reference to the accompanying drawings in which: FIG.\",\n \"1 is a schematic illustration of a healthcare management system in accordance with an embodiment of the invention; FIG.\",\n \"2 is a schematic illustration of patient apparatus in accordance with an embodiment of the invention; FIGS.\",\n \"3 and 4 illustrate look-up tables used in embodiments of the invention; FIGS.\",\n \"5 to 8 are flow diagrams illustrating procedures carried out in accordance with embodiments of the invention; and FIGS.\",\n \"8 to 11 illustrate embodiments of patient apparatus in greater detail.\",\n \"FIG.\",\n \"1 illustrates a healthcare management system arranged in accordance with an embodiment of the invention.\",\n \"The system includes a patient healthcare management system A, a healthcare service provider data management system B, a physician's data management system C, a pharmacy data management system D, and manufacturer data management system E. Patient healthcare management system A includes a medicament dispenser 2 which includes an outer housing which takes a refill 4, containing medicament, which is inserted into a receiving inlet 5 of the dispenser and securely fastened therein.\",\n \"The refill 4 is removable therefrom once the refill is empty, or near to empty, to be replaced by a further refill.\",\n \"The dispenser includes an alphanumeric display 28 for the display of functional data and one or more man machine interface elements, for example buttons 30, for the control of the functions of the dispenser by the patient.\",\n \"The dispenser 2 is adapted for insertion, in a drop-in fashion, into a docking station 6 for the transfer of data between the dispenser 2 and a service provider network server 8, and for the recharging of a battery and/or charge capacitors providing electrical power in the dispenser, in particular its various functions to be described in further detail below.\",\n \"The docking station includes an electricity mains supply cable 9 and, internally, a voltage transformer for the supply of power at a required low voltage.\",\n \"In case of mains supply failure or unavailability the docking station may further include a back-up battery, which may be rechargeable or non-rechargable.\",\n \"The docking station 6 also includes a communications module 10 for communicating with a corresponding communications module 12 on a network interface device 14.The network interface device 14 includes a network interface module 16 for access to a public data communications network 18, such as the Internet.\",\n \"Both the docking station and the network interface device 14 may be located in a patient's home.\",\n \"The communications modules 10, 12 preferably communicate via a radio interface protocol, such as the Bluetooth™ radio protocol, thereby allowing the docking station 6 to be located at a selected point in the patient's home, such as a bedroom or bathroom, remote from the network interface device 14.The network interface module 16 may comprise one or more of a PSTN modem, a DSL modem, a cable modem, a wireless radio network modem.\",\n \"The network interface device may itself take the form of an appropriately configured personal computer workstation, digital television set top box, home communications hub, etc.\",\n \"FIG.\",\n \"2 schematically illustrates the electronic components of the medicament dispenser 2, refill 4 and docking station 6 in greater detail.\",\n \"The dispenser 2 includes a data processor, for example an integrated circuit chip containing a microprocessor 100 and non-volatile memory, in the form of for example ROM 108 and EEPROM 110.Some parts of the memory are reserved for writing to during the manufacture of the dispenser and are, following manufacture, read-only or effectively read-only by virtue of being protected by a secret authentication code, preferably known only by the manufacturer.\",\n \"These read-only parts include a memory portions storing one or more of a unique identifier for the dispenser, a random or pseudo-random password related thereto, one or more type codes for the one or more different types of medicament the dispenser is intended to be used with, a usage lifetime indicator in the form of an expiry date and/or a prescribed maximum number of usages for the dispenser, manufacturing compliance check data and manufacturer details.\",\n \"Other parts are writable and are preferably protected, requiring a predetermined authentication step to have been performed before being capable of being written to, or rewritten.\",\n \"A first set of one or more records are intended to be written initially during dispensing of the dispenser, including one or more of a service provider identifier and/or service provider network address, activation flag, details of an initially prescribed treatment regimen, a version number thereof, and patient name and other patient details.\",\n \"A second set of one or more records are intended to be written initially to by the docking station 6, including records indicating a variation from the original treatment regimen, a version number thereof, and a deactivation flag.\",\n \"A third set of one or more records are intended to be written initially to by the dispenser during use, including compliance monitoring data records indicating when medicament has been taken by the patient and what amount thereof.\",\n \"Some further parts of the memory, containing secret data and/or algorithms, are not externally readable and only for internal use by the microprocessor in performing authentication and encryption procedures.\",\n \"The dispenser 2 comprises external electrical contacts 20, corresponding to contacts in the docking station, for writing data to and reading data from the dispenser, and for charging the battery and/or capacitors of the dispenser.\",\n \"The refill 4 includes a data processor, for example an integrated circuit chip containing a microprocessor 116 and non-volatile memory, in the form of for example ROM 118 and EEPROM 120.Some parts of the memory are reserved for writing to during the manufacture of the refill and are, following manufacture, read-only or effectively read-only by virtue of being protected by a secret authentication code, preferably known only by the manufacturer.\",\n \"These read-only parts include one or more memory portions storing one or more of a unique identifier for the refill, a type code for the stored medicament, a medicament name, an expiry date, an original dosage amount, manufacturing compliance check data and manufacturer details.\",\n \"Other parts are writable and are preferably protected, requiring a predetermined authentication step to have been performed before being capable of being written to, or rewritten.\",\n \"A first set of one or more records are intended to be written initially during dispensing of the refill, including one or more of an activation flag, details of a prescribed treatment regimen, a version number thereof, and patient name and other patient details.\",\n \"A second set of one or more records are intended to be written initially to by the medicament dispenser, including one or more records indicating a current dosage amount remaining.\",\n \"Some further parts of the memory, containing secret data and/or algorithms, are not externally readable and only for internal use by the microprocessor in performing authentication procedures.\",\n \"The refill 4 comprises external electrical contacts 22, corresponding to contacts in the dispenser 2, for writing data to and reading data from the refill.\",\n \"The docking station 6 includes a data processor in the form of for example an integrated circuit chip containing a microprocessor 122 and non-volatile memory, in the form of for example ROM 128 and EEPROM 130.Some parts of the memory are reserved for writing to during the manufacture of the docking station and are, following manufacture, read-only or effectively read-only by virtue of being protected by a secret authentication code, preferably known only by the manufacturer.\",\n \"These read-only parts include one or more memory portions storing one or more of a unique identifier for the docking station, manufacturing compliance check data and manufacturer details.\",\n \"Other parts are subsequently writable.\",\n \"A first set of one or more records are intended to be written to from the dispenser when the dispenser 2 is docked with the docking station, including a medicament name, a current remaining dosage amount, details of a current treatment regimen and patient name and other patient details.\",\n \"These first records contain data to be displayed on a screen 24.Screen 24 is larger than display 28, and is capable of displaying bitmap images, for example logos, still and video pictures, etc, as well as alphanumeric text, responsive to user interaction via an MMI such as buttons 26.A second set of one or more records are intended to store message data received from service provider network server 8, including one or more records for storing data for messages to be shown on screen 24.A third set of one or more records are for storing control messages to be transmitted to dispenser 2, such as control messages for performing authentication and control messages indicating a variation in treatment regiment, indicating a current dosage amount remaining.\",\n \"A fourth set of records are for storing data for transmission to network server 8, including compliance monitoring data received from dispenser 2, and self-test health check data, and self-test health status data records, containing data such as peak flow rate data in the case of a respiratory inhaler or blood glucose level data in the case of an insulin dispenser, which is recorded by a sensor 132 in, or associated with, the docking station.\",\n \"In an alternative, the self-test health check data may be replaced by patient self-assessment data, wherein the patient inputs, via MMI 26, a self-assessed health condition indicator, for example a figure from 1 to 10, indicative of how the patient feels.\",\n \"The patient may also append the data with text to record additional data relating to their state of health.\",\n \"The self-test health check data, condition indicator data and other data are automatically date stamped for subsequent analysis by network server 8 and presentation, for example via a patient-specific Web site page, via charts and other assessment tools.\",\n \"Dispenser display 28, for example an LCD screen, is for displaying information to the patient including the name of the medicament contained in an inserted refill, a current dose remaining, a current time, etc.\",\n \"Reminder messages may also be displayed thereon to indicate when a dose of medicament is due to be taken, in accordance with the current treatment regimen.\",\n \"Such a reminder message may accompanied by a timed alarm indication, given for example by an audible (e.g.\",\n \"tone), kinematic (e.g.\",\n \"vibration) and/or visual (e.g.\",\n \"LED) signal generator.\",\n \"Patients may control display menu and functions by means of a user interface, such as buttons 30.When administering medicine, patient opens outlet 32 and presses a button 30 to initiate the dispensing of a dose, or multiple doses, of medicament.\",\n \"The amount dispensed is controlled by dispenser 2 in accordance with a current treatment regimen stored in the memory of the dispenser 2 and/or the refill 4, along with the current date/time and/or stage of treatment.\",\n \"Dispenser 2 further comprises a dispensing actuator 102, for the automated dispensing of a dose, or multiple doses of medicament.\",\n \"In one embodiment, the medicament is held in refill 4 in powder, liquid and/or gaseous form and a dispensing actuator is adapted to meter out a required dose from refill 4.In another embodiment, refill 4 contains medicament in the form of elements containing discrete amounts of medicament, for example tablets, capsules, powder compartments, etc.\",\n \"and dispensing actuator is adapted to automatically dispense one or more discrete doses of medicament.\",\n \"Dispensing may be initiated by the patient, for example by means of MMI 30.Dispenser 2 further comprises dispensing sensor 104, for sensing the act of dispensing and recording, as monitoring data, the amount, time and date when dispensing occurs.\",\n \"Clock 106 is used for time monitoring and recordal in the dispenser electronic system.\",\n \"Data interface 22, consisting of two sets of corresponding electrical contacts, is used for the transfer of data between the refill 4 and the dispenser 2.In an alternative embodiment, the interface consists of a contactless radio interface between a passive tag in the refill and an active reader in the dispenser.\",\n \"Data interface 114, consisting of two sets of corresponding electrical contacts, is used for the transfer of data between the docking station 6 and the dispenser 2.In an alternative embodiment, the interface consists of a contactless radio interface, for example a Bluetooth™ radio interface which is operable when dispenser is placed in the proximity of the docking station 6.Power interface 114 consists of two sets of corresponding electrical contacts for the transfer of electrical power between docking station power transformer 134, supplied by mains power source connection 9, and rechargeable battery 112 in the dispenser 2, which acts as a power source for all electronic components in the dispenser 2 and the refill 4.Transformer 134 acts as a power source for all electronic components in the docking station 6.Docking station 6 may also include a rechargeable back-up battery in case of mains power failure.\",\n \"Docking station 6 includes a data processing system including a microprocessor 122, non-volatile memory in the form of ROM 128 and EEPROM 130, display 24, man machine interface elements 26, radio interface module 10, including radio antenna 126, a mains power source connection 9, and self-test module 132.Self-test module 132 includes a patient health sensor, such as a blood glucose level sensor, peak flow sensor, etc, and may be detachably connectable to the docking station.\",\n \"In an alternative embodiment, self-test module 132 is included in dispenser 2, or may be detachably connectable to the dispenser 2.Service provider network server 8 is connected to patient record database 34, which stores long-term patient health records to be used by the health system for the treatment of patients under its care.\",\n \"These records store historical patient health and treatment data, including prescription data, treatment regimen compliance data, and health check status data.\",\n \"Service provider network server includes analysis and reporting functions for operating on data contained in the patient health records and for transmitting messages and reports to other healthcare management systems including patient healthcare management systems, physicians' data management systems and pharmacy data management systems.\",\n \"Manufacturer data management system E includes a manufacturing system server 36, a data writing terminal 38 for writing data to a dispenser 40 during manufacture, and a data writing terminal 40 for writing data to refill 4 during manufacture.\",\n \"During manufacture, the dispenser memory 108, 110 is written to contain permanent data in the form of one or more of a unique identifier for the dispenser, a random or pseudo-random password, one or more type codes for the one or more different types of medicament the dispenser is intended to be used with, a usage lifetime indicator in the form of an expiry date and/or a maximum number of usages for the dispenser, a geographical region name and/or code, manufacturing compliance check data and manufacturer details.\",\n \"The dispenser memory is also written to with secret data and/or algorithms, which are not externally readable and only for internal use by the dispenser microprocessor in performing authentication and encryption procedures.\",\n \"This secret data includes a secret authentication key, which may be of a shared secret type of a private key of a public/private key combination.\",\n \"The data may also include a public key for one or more service providers, or a trusted third party providing service provider public key details, to authenticate and/or decrypt control messages received from service provider network server 8, which are encrypted and/or digitally signed by the server 8 using its private key and/or the appropriate shared secret key before transmission.\",\n \"The data may also include a public key for one or more manufacturers, or a trusted third party providing manufacturer public key details, to authenticate and/or decrypt data received from refill 4, which are encrypted and/or digitally signed by the manufacturer or on the refill before transmission.\",\n \"Following manufacture, the data written to the dispenser memory, including in particular the unique identifier and the corresponding password and/or shared secret key and/or public key, are transmitted in a secure fashion to service provider network server 8 and stored therein awaiting dispensing of the dispenser 2 to occur.\",\n \"The dispensers are batched and distributed in batches to pharmacies for subsequent dispensing to patients.\",\n \"During manufacture, the refill memory 118, 120 is written to contain permanent data in the form of one or more of a unique identifier for the refill, a type code for the stored medicament, a medicament name, an expiry date, an original dosage amount, a geographical region name and/or code, manufacturing compliance check data and manufacturer details.\",\n \"The permanent data also includes a digital signature, generated by manufacturer using a secret authenticating key, of the unique identifier and/or other data, which is added to the refill memory.\",\n \"The refill memory may also be written to with secret data and/or algorithms, which are not externally readable and only for internal use by the refill microprocessor in performing authentication procedures.\",\n \"This secret data may include a secret authentication key, which may be of a shared secret type of a private key of a public/private key combination.\",\n \"The data may also, or alternatively, include a public key for the service provider, to authenticate and/or decrypt control messages received from service provider network server 8, which are encrypted and/or digitally signed by the server 8 using its private key and/or the appropriate shared secret key before transmission.\",\n \"Following manufacture, the data written to the refill memory, including in particular the unique identifier and the corresponding digital signature, shared secret key and/or public key, are transmitted in a secure fashion to service provider network server 8 and stored therein awaiting dispensing of the refill 4 to occur.\",\n \"The refills are batched and distributed in batches to pharmacies for subsequent dispensing to patients.\",\n \"Patient system A also contains a smart card 42 for identifying the patient and storing patient healthcare details, which preferably are replicated in patient record database 34, including general health data, prescription data, allergy data, etc.\",\n \"Smart card 42 includes electrical contacts 43 whereby a smart card reader/writer accesses data records held on the smart card.\",\n \"Physician data management system C includes a workstation personal computer 44, a smart card reader/writer 46 and a dispenser reader/writer 48.The dispenser reader/writer 48 may be a docking station similar to the patient docking station 6.On visiting a physician, a patient preferably carries their smart card 42.The smart card is inserted in reader/writer 46 to allow the physician to access patient data held thereon and/or patient data held in patient record database 34.The data is accessed via workstation 44.Each physician has a personal username and password providing the physician with authority to access such data, which details are verified by network server 8 before such access is granted.\",\n \"Following consultation, the physician may issue a digital prescription, which is written to smart card 42 and/or patient record database 34.Pharmacy data management system D includes a server 50, a workstation personal computer 52, a dispenser reader/writer 54, a refill reader/writer 56 and a smart card reader/writer 58.The dispenser reader/writer 54 may be a docking station similar to the patient docking station 6.When a patient visits the pharmacy to obtain supplies, the patient carries their smart card 42, which is inserted in smart card reader 58 in order to identify the patient and the corresponding prescription.\",\n \"The patient data is accessed via workstation 52.Each pharmacist has a personal username and password providing the pharmacist with authority to access such data, which details are verified by network server 8 before such access is granted.\",\n \"The patient identity is transmitted to network server 8 in order to access and/or the prescription data.\",\n \"The patient's data record indicates whether the patient currently holds a dispenser appropriate for the prescribed medicament.\",\n \"If not, as in the case of a newly diagnosed patient, the network server instructs pharmacist system to dispense a new dispenser 2.In order to dispense a new dispenser 2, the pharmacist retrieves the appropriate dispenser from the pharmacy stores and inserts the dispenser into dispenser reader/writer 54.The reader/writer 54 reads the dispenser's unique identity and transmits same to network server 8.FIG.\",\n \"3 shows a look-up table held database in 34, associating each unique dispenser ID, and the associated data fields, corresponding to the original data held in the dispenser memory, provided by manufacturer (illustrated as Field #1 etc.)\",\n \"with a patient ID following dispensing thereof.\",\n \"The reader/writer is then used to write data received from network server 8, containing one or more of a service provider identifier and/or service provider network address, activation message, details of an initially prescribed treatment regimen, a version number thereof, patient ID and name and other patient details, and a geographical region code, to the dispenser memory.\",\n \"The written data is preferably encrypted and/or signed by the network server 8, thereby indicating to the dispenser that a pharmacist having the necessary authority is performing the dispensing operation.\",\n \"Once activated, the dispenser 2 is ready for use with an appropriate refill 4.In order to dispense a new refill 4, the pharmacist retrieves the appropriate refill from the pharmacy stores and inserts the dispenser into dispenser reader/writer 54.The reader/writer 54 reads the refill's unique identity and transmits same to network server 8.FIG.\",\n \"2 shows a look-up table held database in 34, associating each unique refill ID, and the associated data fields, corresponding to those originally held in the refill memory, provided by manufacturer (illustrated as Field #1 etc.)\",\n \"with a patient ID following dispensing thereof.\",\n \"The reader/writer is then used to write data provided by network server 8, containing one or more of an activation message, details of a prescribed treatment regimen, a version number thereof, and patient ID, name and other patient details, and a geographical region code, to the refill memory.\",\n \"The written data is preferably encrypted and/or signed by the network server 8, thereby indicating to the refill that a pharmacist having the necessary authority is performing the dispensing operation.\",\n \"Once activated, the refill 4 is ready for use with an appropriate dispenser 2.In one embodiment, the pharmacy procedures include reading the data stored on the dispenser and/or refill prior to dispensing, and modifying the dispensing operation accordingly, either in the pharmacy system alone or in combination with the network server system.\",\n \"For example, the pharmacy may be prevented from dispensing a device having a geographical region name and/or code which does not correspond for refills and dispensers, or if the location of the pharmacy does not correspond therewith.\",\n \"The look-up tables shown in FIGS.\",\n \"3 and 4 are used for the correct identification of a patient by means of the corresponding dispenser and/or refill ID's.\",\n \"When data is transferred from a dispenser 2 and/or a refill 4 to network server 8, the data includes the device ID and/or the refill ID.\",\n \"Thereby, the patient is identified without requiring the patient to input username and/or password details each time data is sent to the network server 8.Furthermore, the associated authentication procedures may be performed by means of the devices themselves, rather than the patient having to identify and carry out authentication each time sensitive data is transferred.\",\n \"The corresponding shared authentication data, such as shared secret keys, held on both the devices and in the device-related parameter fields illustrated in FIGS.\",\n \"3 and 4, may be uniquely correlated by means of the look-up tables to the identity of the patient to whom the device was prescribed.\",\n \"In an alternative embodiment, the unique patient ID is written at the pharmacist onto the dispenser and/or refill during dispensing, and the patient ID is sent with any data sent from dispenser to network server 8.However, in this case a device ID, which may be unique amongst all devices, or unique only in combination with the patient ID, is preferably also sent to allow the identification of the device currently being used by the patient at the network server 8.In any case, the data transmitted to network server 8 contains a unique identity, preferably device ID, whereby the corresponding patient record may be accessed.\",\n \"FIG.\",\n \"5 is a flow diagram illustrating procedures carried out in the dispenser 2 on insertion of a refill 4.On insertion of the pharmacy-dispensed refill 4 into the pharmacy-dispensed dispenser 2, the dispenser reads all available data held in refill memory, step 200.The dispenser then checks that no conflicts exist between the data read and that stored in its memory, steps 202 and 204, including ensuring that the refill has a matching patient ID, acceptable medicament type code, acceptable geographical region code, etc.\",\n \"Furthermore, dispenser 2 checks that the any digital signature accompanying the parameter data held in refill is correct.\",\n \"This check is carried out, in one embodiment, by reading the appropriate signature and decrypting it with the public key of the network server 8 to ensure that it corresponds with the original data, for example the unique identity of the refill 4, to which the signature is attached.\",\n \"In other embodiments, other cryptographic methods are used to check the authenticity of the refill by means of public/private key or shared secret key encryption/decryption methods.\",\n \"If a conflict is found, a conflict indicator signal is generated.\",\n \"If no conflict is found, the dispenser checks if there are any updates to be received from the refill.\",\n \"For example, the refill may contain a more recent treatment regimen, which may be detected by means of a later version number.\",\n \"If an update is detected, it is first authenticated, by checking the accompanying digital signature, before updating the appropriate parameters in its memory.\",\n \"If the update authentication fails, a conflict indicator signal is generated.\",\n \"Otherwise, following any update, the dispenser 2 begins a normal treatment regimen in accordance with data held on the dispenser and/or the refill 4.If a conflict indicator signal is generated, the dispenser display 28 is activated to show an appropriate error message, for example “Refill not authorised: see pharmacist”, step 206.The dispenser 2 also inhibits any reminder programme which would otherwise have been followed, step 208.On generation of the conflict indicator signal, the dispenser may, in one embodiment, write a deactivation flag in the dispenser or refill memory to prevent the refill and/or the dispenser being used, at least until the conflict is resolved by means of an update message received from server 8.Alternatively, the dispenser may still be used in manual override mode to dispense medicament from the refill, even though the data management functions are not made available.\",\n \"Further alternatively, the dispenser may continue to operate normally but store the conflict data to transmit same to network server 8 when the dispenser is next docked.\",\n \"The action taken by dispenser may depend upon the nature of the conflict found.\",\n \"For example, if the refill contains the wrong type of medicine, the refill may be fully deactivated.\",\n \"On the other hand, if the refill was dispensed to a different patient, but contains the correct type of medicament, the dispenser may be used as normal, with the conflict data being sent to network server 8 to be written to the patient record in order that a healthcare professional, such as the physician or pharmacist, may be made aware of the conflict when the patient record is next accessed.\",\n \"The dispenser then senses whether a docking connection is made to docking port 6, for example by means of a data read signal being received from docking port 6, at which point an message indicating the detected conflict is sent, via docking station 6, to network server 8, steps 210 and 212.Network server 8 may used the conflict data for monitoring purposes, and inform the appropriate healthcare professional as and when required of the detected conflict.\",\n \"For example, when patient next visits a pharmacist, the pharmacist will be able to access, via the patient record, the conflict data and indicated the nature of the problem to the patient.\",\n \"If the conflict is an internal conflict caused by an error in the network server system 8, the conflict may be resolved and updates may be sent to all other affected dispensers to avoid the conflict occurring unnecessarily elsewhere.\",\n \"Preferably, the dispenser may still be used in manual override mode to dispense medicament from the refill, even thought the data management functions are not made available.\",\n \"In any case, when a conflict is found, conflict data indicating the cause of the conflict, e.g.\",\n \"both the data derived from the refill and the corresponding, incompatible, data held on dispenser, is transmitted to network server 8.Network server 8 analyses the nature of the conflict and can take remedial action, such as writing corresponding data to patient record for perusal by a healthcare professional having access to the system, transmitting informational messages to the patient via the patient docking station, and/or transmitting update messages to the dispenser via the docking stations to resolve the conflict by updating one or more fields of the dispenser memory as appropriate.\",\n \"If no conflict is found, the unique identity of the refill and/or other details held on the refill are written to a record in the dispenser memory for transfer, via docking station 6, to the network server 8 when the dispenser is next docked.\",\n \"Such data, derived from the refill, is then stored in the appropriate patient record by the network server 8, to provide a medication history, including details of the refill container used, for the patient.\",\n \"Furthermore, in an alternative embodiment, any conflict checks are carried out by network server.\",\n \"In this embodiment, the current refill ID and device ID are preferably transmitted to network server 8, and the corresponding parameter data, held in the lookup tables illustrated in FIGS.\",\n \"3 and 4, is analysed by network server 8 to determine whether a conflict exists.\",\n \"Alternatively, one or more of the other parameter data items held in the dispenser memory and/or the refill memory may be transmitted to network server 8.If a conflict is found, network server 8 analyses the nature of the conflict and can take remedial action, such as writing corresponding data to patient record for perusal by a healthcare professional having access to the system, transmitting informational messages to the patient via the patient docking station, and/or transmitting update messages to the dispenser via the docking stations to alter the functioning of the dispenser, to disable the dispenser until a new refill has been inserted, and/or to resolve the conflict by updating one or more of the previously incompatible fields of the dispenser memory as appropriate.\",\n \"FIG.\",\n \"6 illustrates a treatment procedure followed by dispenser 2 when loaded with a correct refill 4.The dispenser holds treatment regimen data from which it is determined when a medicament dose is due to be taken by the patient, step 300.A period is defined when the dose is defined to be correctly taken, by means of a start date/time and an end/date time, which may be defined relative to a last taken medication or in accordance with an absolute timing policy.\",\n \"If the patient activates the dispenser prior to the start date/time, step 302, the dispenser acts in accordance with an early dispensing procedure, step 304, which is different to the normal dispensing procedure.\",\n \"This may be to at least initially inhibit dispensing and display a message to the patient indicating that it is too early to take another dose.\",\n \"An option may at this stage be given to allow the patient to confirm that a dose is required, in response to which the dispenser conducts dispensing as required.\",\n \"Alternatively, dispenser may initially dispense the dose at the requested time, and update the subsequent reminder schedule to cancel the upcoming reminder and bring forward the subsequent reminders.\",\n \"When a dosing reminder is due, the dispenser 2 emits a dosing reminder signal, step 306.The signal may be an audio, visual and/or kinematic signal, and may vary over time depending on whether dosing is carried out by the patient, for example to increase in intensity over time.\",\n \"When patient actuates the dispenser to dispense a dose of medicament, step 310, the dispenser determines whether the dispensing operation is later than the predefined end date/time for the normal dosing period, step 310.If so, a late dispensing procedure, different to a normal dispensing procedure, is carried out, step 318.This may consist of indicating to the patient that the dose is being taken late and adjusting the subsequent reminder schedule accordingly, to delay subsequent reminders in accordance with the delay.\",\n \"If within the normal dosing period, the normal dosage data is accessed, step 312, and the appropriate dosage is dispensed, step 314.Following all dispensing procedures, although only shown in relation to normal dispensing procedure, the monitoring data held in dispenser, subsequently to be sent to network server 8, is updated to store details of the latest dispensing event, including date/time, amount dispensed, and any other pertinent data such as patient health data (e.g.\",\n \"peak flow rate) detected during the dispensing operation.\",\n \"FIG.\",\n \"7 illustrates a procedure carried out by docking station 6 when a dispenser is docked therein.\",\n \"Note that in this preferred embodiment, the docking station 6 may be used with multiple different dispensers, including multiple dispensers owned by the same patient and/or multiple dispensers owned by different patients.\",\n \"On receiving a dispenser 2, the docking station 6 first reads the patient name and medicament name, along with other pertinent data such as number of doses remaining, etc., and displays these details to the patient, steps 400, 402, via display 24.The patient may scroll through the relevant data held on dispenser by means of an MMI interaction.\",\n \"Next, the docking station reads other stored data from the dispenser, including the dispenser identity, the refill identity, any conflict data resulting from a conflict check conducted by dispenser 2, and dispensing monitoring data stored when the patient has dispensed medication from the dispenser 2.This stored data may be provided by the dispenser in encrypted form, for example encrypted using its stored secret key, for transmission by the docking station in encrypted form, step 406, and decryption by the network server 8 with reference to the corresponding decryption key held in the details held for the identified dispenser.\",\n \"Furthermore, authentication data, in the form of a digital signature produced by the dispenser 2 and/or the refill 4, may also be transmitted for checking by network server.\",\n \"The docking station may form a suitable data connection, for example a TCP/IP socket connection, to the network server 8.The connection between the docking station 6 and the network server 8 may in any case be arranged to be secure, for example via secure socket layer (SSL) interactions.\",\n \"The data sent by docking station is analysed at the network server 8 and one or more response messages are generated and sent to be received by docking station, step 408.These messages may be structured as extensible markup language (XML) documents.\",\n \"The responses preferably comprise data to be displayed to user via the display screen 24 of the docking station, and update data to be passed by docking station.\",\n \"Docking station 6 parses the response data, step 410, displays the display content, step 412, and transmits any update messages to the dispenser 2, step 414.The update messages are preferably encrypted by network server 8 using the corresponding encryption key, for example the dispenser's shared secret key or the public key of the dispenser, and/or contain a further authentication element, such as a digital signature authenticating the data as having been generated by, or with the authority of network server 8.FIG.\",\n \"8 illustrates steps taken by dispenser 8 on receipt of an update message via docking station 6, step 500.Initially, the dispenser 2 conducts decryption of the data using a secret key stored in the dispenser memory, step 502.Next, the dispenser checks the accompanying digital signature, step 502.If either of these steps fails, the update message cannot be authenticated, step 506 by dispenser 2 and an error message is displayed, step 510.An error message may also be sent to network server 8, to allow a further, corrected, update to be sent.\",\n \"If the update is authenticated by the dispenser, step 506, the corresponding data fields are updated in the dispenser memory and/or the refill memory, step 508.It is to be noted that any one or more of the writable fields of the dispenser and/or the refill, such as any of those previously mentioned, may be updated in this way.\",\n \"If the dispenser 2 and/or the refill 4 fails to authenticate to network server 8, the network server can send an appropriate message to the docking station, such as “Authentication failed: please visit pharmacy”.\",\n \"In a subsequent physician visit, if the patient has brought their dispenser 2, and the dispenser is inserted into reader/writer 48 in order for the most recent compliance and health status data to be downloaded to network server 8.The dispenser's unique identity is transmitted to network server 8, along with authentication data in the form of the corresponding password and/or, if stronger authentication is required, an encrypted response to a challenge message sent by the server 8, which response is verified by server 8 in order to validate the dispenser identity.\",\n \"The corresponding patient identity is looked up by network server in order to access the correct patient record.\",\n \"Not only does physician have access to patient data, but also the analysis and reporting functions provided by network server 8 following the period of patient compliance and health monitoring provided by dispenser 2.Following consultation with the network server functions and the patient, the physician may vary the treatment regimen, by input to workstation 44 which results in an update procedure whereby network server 8 transmits data to be written to the dispenser by reader/writer 48.The transmitted data is encrypted and/or digitally signed by the network server, using the appropriate secret and/or private key for the dispenser 2.On receipt, the dispenser decrypts and/or authenticates the control data and, if authentication is successful, updates its records accordingly.\",\n \"The physician may also prescribe more or a new medicament, in which case a digital prescription is written to smart card 42 and/or to patient record database 46.If on docking with docking station 6, either the docking station or the network server 8 establishes that the patient is due for another prescription, a message may be displayed to the patient inquiring whether a further prescription should be ordered.\",\n \"If the patient replies in the affirmative, a represcription order placed in the patient record, or is transmitted to a selected pharmacist, and the patient may pick up the appropriate refill by presenting at the pharmacist with their patient smart card 42.The network server-based system also monitors the lifetimes of dispensers and/or refills.\",\n \"Namely, associated with each of the dispensers and refills, by means of parameter data held against the device identity in the look-up tables illustrated in FIG.\",\n \"4, is an expiry date for the device, and when the device is due to expire, a suitable message may be sent to docking station instructing the patient to order the prescription of a new device.\",\n \"On expiry an update message may be sent to the dispenser via the docking station to disable dispensing.\",\n \"The lifetime of the dispenser may be based solely on an expiry date, a usage amount (e.g.\",\n \"maximum number of allowed uses), or both.\",\n \"In any case, such monitoring and messaging is useful in order to inhibit the at least inconvenient failure of any of the devices used by the patient.\",\n \"In the above-described embodiments, the patient is identified primarily by means of the dispenser and/or the refill which they have been dispensed by a registered pharmacist.\",\n \"Additional identity checks may be carried out by the system.\",\n \"For example, the dispenser 2 or the docking station 6 may include a Biometric sensor such as a fingerprint sensor for positively identifying the patient before any actions based upon the patient identity are carried out.\",\n \"In different embodiments of the invention, the dispenser is typically shaped to define a cavity within which the refill is receivable.\",\n \"The dispenser and/or refill may be further shaped with grooves, indentations or other shaping or surface details to define a slidingly lockable/releasable relationship between the dispenser and the refill.\",\n \"Accordingly, the dispenser may have a lockable cover which is releasable to expose the cavity within which the refill is held.\",\n \"FIGS.\",\n \"9a and 9b show a metered dose inhaler comprising a dispenser in the form of a tubular actuator housing 601 shaped for receipt of an aerosol refill 602.The actuator housing is open at one end and is closed at the other.\",\n \"An outlet 603 leads laterally from the closed end of the housing 601.In the embodiment illustrated, the outlet 603 is in the form of a mouthpiece intended for insertion into the mouth of the patient but it may, if desired, be designed as a nozzle for insertion into the patient's nostril.\",\n \"The aerosol refill 2 has an outlet valve 604 at one end.\",\n \"This valve acts as a release means for release of a measured dose from the aerosol refill.\",\n \"The release means is actuable by inward movement of the valve 4 relative to the aerosol refill 602.The metered dose inhaler of FIGS.\",\n \"9a and 9b includes an electronic data management system 610, including all electronic components and electrical connections, and is adapted for docking with a corresponding docking station, as described above in detail, comprised within an extended part 606 of the housing 6.A visual display (not visible) allows for display of information from the electronic data management system 610 to the patient.\",\n \"The electronic data management system connects to a sensor (not visible) for sensing the breathing pattern of the patient and an actuator (not visible) for actuating the release of aerosol from the refill 2.A data module in the form of a chip 620 is mounted on the side of the aerosol refill 602.Corresponding read/write contacts 630 for the chip 620 are moulded into the internal surface of the dispenser housing 601.In an alternative embodiment the data module 620 comprises a radiofrequency identification tag and the read/write module 630 is capable of reading data therefrom and writing data thereto by the use of interrogating radiofrequency energy.\",\n \"In another embodiment the data module 620 comprises a magnetic label and the read/write module 30 is capable of reading data therefrom and writing data thereto by the use of interrogating magnetic field energy.\",\n \"FIG.\",\n \"10 shows a schematic representation of a breath-operable medicament dispensing system, which is an embodiment of the system shown in FIGS.\",\n \"9a and 9b.\",\n \"The system comprises a metered dose inhaler similar to that shown in FIGS.\",\n \"9a and 9b, comprising tubular housing 710 having a dispensing outlet 712 in the form of a mouthpiece.\",\n \"Within the housing 710 sits aerosol refill 720 which has a valve dispensing mechanism 722 in the form of a slide valve.\",\n \"Valve stem 724 is supported by valve support 714.Outlet passage 716 is provided in the support 714 to enable passage of dispensed dose to the dispensing outlet 712.It may be seen that the upper part of the aerosol refill 720 abuts refill seat 730.The refill seat 730 comprises an insulating portion 732 which directly contacts the aerosol refill 720 and an upper conducting portion 734 (e.g.\",\n \"comprised of aluminium).\",\n \"It may also be seen that the valve support 774 connects with conducting valve seat 740.Plural shape memory alloy wires 750a, 750b connect the conducting portion 734 of the refill seat 730 to the conducting valve seat 740.The plural wires 750a, 750b comprise a nickel-titanium alloy which contracts in response to electrical current flow therethrough.\",\n \"It may thus, be appreciated that on passage of electrical current through the plural wires 750a, 750b the refill seat 730 and valve seat 740 will be drawn towards each other as the wires 750a, 750b contract.\",\n \"Actuation of the valve dispensing mechanism 722 and dispensing of medicament dose will thereby result.\",\n \"Control of electrical current flow to the refill seat 730, valve seat 740 and wires 750a, 750b is achievable using the illustrated circuitry.\",\n \"Refill seat 730 and valve seat 740 connect to actuation circuit 760 which includes a high current power supply 762 (e.g.\",\n \"a voltaic cell or battery of voltaic cells) and a switch 764 in the form of a solid state relay.\",\n \"The solid state relay 764 itself connects with control circuitry including a micro-controller 770 having an independent power supply 772.The micro-controller 770 itself connects with pressure transducer 780 which has an input in the form of a pressure tube 782 located within the dispensing outlet 772 of the inhaler housing 770.It may be appreciated that current flow to the refill seat 730, valve seat 740 and wires 750a, 750b, and hence actuation of the valve dispensing mechanism may be achievable as follows.\",\n \"The patient inhales through the mouthpiece 772 resulting in a change in pressure within the housing 770 and pressure tube 782.The change in pressure is detected by the pressure transducer 780 which sends a signal to the micro-controller 770.The micro-controller 770, in turn sends a switching signal to the solid state relay 764 which results in closing of the actuation circuit and electrical current flow therethrough.\",\n \"The resulting contraction of the shape memory alloy wires 750a, 750b causes actuation of the valve dispensing mechanism 722 and hence, dispensing of medicament to the inhaling patient.\",\n \"It may also be seen in FIG.\",\n \"10 that the micro-controller 770 is connected to a display 774 for display of information to the patient and also with a computer interface 776 for exchange of data therewith.\",\n \"All circuitry and components thereof including the power supplies 762, 772 and display 774 may be arranged to be present on the housing 770 such that the system is in the form of a discrete, hand-held device.\",\n \"FIG.\",\n \"11 shows a medicament dispenser 800 containing a refill cartridge 802 in accord with a further embodiment of the present invention.\",\n \"The medicament refill cartridge 802 comprises a flexible strip 804 defining a plurality of pockets each of which contains a discrete dose of medicament.\",\n \"The doses may be in the form of tablets, capsules, doses of powder which can be inhaled, etc.\",\n \"The strip 804 comprises a base sheet in which blisters are formed to define the pockets and a lid sheet which is hermetically sealed to the base sheet except in the region of the blisters in such a manner that the lid sheet and the base sheet can be peeled apart.\",\n \"The lid and base sheets are each preferably formed of a plastics/aluminium laminate and are preferably adhered to one another by heat sealing.\",\n \"FIG.\",\n \"11 shows the internal mechanism of medicament dispenser 800 in the situation where the majority of the pockets of the refill strip 804 are still filled with medicament.\",\n \"The internal mechanism comprises an index wheel 806 and a lid-winding spool 808 for winding the used portion of the lid sheet.\",\n \"The index wheel 806 has a plurality of recesses extending parallel with the axis of the wheel.\",\n \"The recesses are spaced at a pitch which is equal to the distance between the centre lines of adjacent pockets in the refill strip 804.The refill 802 includes an internal area for the medicament strip 802 to be coiled in prior to use of the doses contained inside it.\",\n \"The dispenser comprises an area where the used base and lid of the medicament carrier strip 804 is collected.\",\n \"After complete usage, the used base may be rewound back into refill automatically prior to removal of the refill.\",\n \"The dispensing system 800, 802, includes an electronic data management system 810, including all electronic components and electrical connections, as described above in detail, and is adapted for docking with a corresponding docking station as described in detail above.\",\n \"The refill 802 includes a data module 812 in the form of a chip, as described above, which interfaces with the data management system 810 when the refill 802 is inserted in the dispenser 800, via a hinged insertion door 814, which is secured in place by means of a catch 816.On insertion of the refill, the lid sheet is wrapped around spool 808 and the base sheet, containing the pockets of medicament, is inserted in transport passage 818 of the dispenser.\",\n \"Spool 808 and/or index wheel 806 are revolved, by means of a motor controlled by data management system 810 a predetermined amount for each dispensing operation.\",\n \"As the dispensing operation occurs, a new dose of medicament is presented at outlet 820 for consumption by the patient.\",\n \"Medicament dispensers of the present invention are in one aspect suitable for dispensing medicament for the treatment of respiratory disorders such as disorders of the lungs and bronchial tracts including asthma and chronic obstructive pulmonary disorder (COPD).\",\n \"Appropriate medicaments may thus be selected from, for example, analgesics, e.g., codeine, dihydromorphine, ergotamine, fentanyl or morphine; anginal preparations, e.g., diltiazem; antiallergics, e.g., cromoglycate (e.g.\",\n \"s the sodium salt), ketotifen or nedocromil (e.g.\",\n \"as the sodium salt); antiinfectives e.g., cephalosporins, penicillins, streptomycin, sulphonamides, tetracyclines and pentamidine; antihistamines, e.g., methapyrilene; anti-inflammatories, e.g., beclomethasone (e.g.\",\n \"as the dipropionate ester), fluticasone (e.g.\",\n \"as the propionate ester), flunisolide, budesonide, rofleponide, mometasone e.g.\",\n \"as the furoate ester), ciclesonide, triamcinolone (e.g.\",\n \"as the acetonide) or 6α,9α-difluoro-11β-hydroxy-16α-methyl-3-oxo-17α-propionyloxy-androsta-1,4-diene-17β-carbothioic acid S-(2-oxo-tetrahydro-furan-3-yl) ester; antitussives, e.g., noscapine; bronchodilators, e.g., albuterol (e.g.\",\n \"as free base or sulphate), salmeterol (e.g.\",\n \"as xinafoate), ephedrine, adrenaline, fenoterol (e.g.\",\n \"as hydrobromide), formoterol (e.g.\",\n \"as fumarate), isoprenaline, metaproterenol, phenylephrine, phenylpropanolamine, pirbuterol (e.g.\",\n \"as acetate), reproterol (e.g.\",\n \"as hydrochloride), rimiterol, terbutaline (e.g.\",\n \"as sulphate), isoetharine, tulobuterol or 4-hydroxy-7-[2-[[2-[[3-(2-phenylethoxy)propyl]sulfonyl]ethyl]amino]-ethyl-2(3H)-benzothiazolone; adenosine 2a agonists, e.g.\",\n \"2R,3R,4S,5R)-2-[6-Amino-2-(1S-hydroxymethyl-2-phenyl-ethylamino)-purin-9-yl]-5-(2-ethyl-2H-tetrazol-5-yl)-tetrahydro-furan-3,4-diol (e.g.\",\n \"as maleate); α4 integrin inhibitors e.g.\",\n \"(2S)-3-[4-({[4-(aminocarbonyl)-1-piperidinyl]carbonyl}oxy)-phenyl]-2-[((2S)-4-methyl-2-{[2-(2-methylphenoxy)acetyl]amino}pentanoyl)-amino]propanoic acid (e.g.\",\n \"as free acid or potassium salt), diuretics, e.g., amiloride; anticholinergics, e.g., ipratropium (e.g.\",\n \"as bromide), tiotropium, atropine or oxitropium; hormones, e.g., cortisone, hydrocortisone or prednisolone; xanthines, e.g., aminophylline, choline theophyllinate, lysine theophyllinate or theophylline; therapeutic proteins and peptides, e.g., insulin or glucagon; vaccines, diagnostics, and gene therapies.\",\n \"It will be clear to a person skilled in the art that, where appropriate, the medicaments may be used in the form of salts, (e.g., as alkali metal or amine salts or as acid addition salts) or as esters (e.g., lower alkyl esters) or as solvates (e.g., hydrates) to optimise the activity and/or stability of the medicament.\",\n \"Preferred medicaments are selected from albuterol, salmeterol, fluticasone propionate and beclomethasone dipropionate and salts or solvates thereof, e.g., the sulphate of albuterol and the xinafoate of salmeterol.\",\n \"Medicaments can also be delivered in combinations.\",\n \"Preferred formulations containing combinations of active ingredients contain salbutamol (e.g., as the free base or the sulphate salt) or salmeterol (e.g., as the xinafoate salt) or formoterol (e.g.\",\n \"as the fumarate salt) in combination with an anti-inflammatory steroid such as a beclomethasone ester (e.g., the dipropionate) or a fluticasone ester (e.g., the propionate) or budesonide.\",\n \"A particularly preferred combination is a combination of fluticasone propionate and salmeterol, or a salt thereof particularly the xinafoate salt).\",\n \"A further combination of particular interest is budesonide and formoterol (e.g.\",\n \"as the fumarate salt).\",\n \"The medicament dispenser of the present invention in other embodiments comprises a syringe for the delivery of injectable medicament to a patient.\",\n \"Traditional syringes rely on puncturing of the patient's skin by a hollow needle through which the injectable medicament (in solution or suspension form) is delivered to the muscle or tissue of the patient.\",\n \"Recently developed needleless systems for the delivery of injectables employ high velocity injection of particle formulated drugs or vaccine through the skin and into any physically accessible tissue.\",\n \"Other needleless systems employ similar high velocity injection of drug or vaccine coated on to a suitable carrier particle.\",\n \"It will be understood that the present disclosure is for the purpose of illustration only and the invention extends to modifications, variations and improvements thereto.\",\n \"In alternative embodiments, there is no docking station situated in the patient home or other environment.\",\n \"However a docking station 6 as described may be provided in a health care provider location, such as in patient clinic, physician's surgery or pharmacist.\",\n \"In this, and other alternative embodiments, the dispenser contains a sealed long-life battery.\",\n \"In the above embodiments, the service provider authentication and decryption procedures for updates messages are carried out by the dispenser 2.In an alternative embodiment, the authentication and/or decryption is performed by the docking station 6.In the above embodiments, the refill is filled with medicament at the site of manufacture.\",\n \"In an alternative embodiment, the refill may be filled in pharmacy, and its memory written to with data identifying its contents and expiry date therefore.\",\n \"In the above embodiments, authentication is performed by means of encryption/decryption and by means of digital signature checking.\",\n \"Other known methods of authentication may also, or alternatively be used.\",\n \"Herein the term digital signature is intended to include digital certificates, which also provide means of authentication.\",\n \"In the above embodiments, the docking station must first establish a connection with the network server in order for update and display messages to be sent to the patient.\",\n \"Other means of delivery of messages, in particular push messages sent by the server at a time determined by the server, may be sent in other formats, for example by means of cellular telephony SMS messages, and to other devices, such as cellular radio devices.\",\n \"In this description, and in the following claims, the devices involved in the present invention include a dispenser and a refill.\",\n \"It is to be noted that the dispenser is not limited to a device comprising a medicament outlet; the outlet may be provided on the refill.\",\n \"The dispenser is a device containing elements for the control of a dispensing operation in which medicament, originally contained in the refill, is dispensed.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473899"}}},{"rowIdx":762,"cells":{"text":{"kind":"list like","value":[["Method for pressure-and flow-preventive fixing of pipes in a well","A method for pressure- and flow-prevetive fixing of well pipes, preferably liners and casings (14, 54, 60, 66) in a well when drilling the well, wherein the method also may be used in a completed well in order to place a pressure- and flow-preventive barrier in an annulus (16) surrounded by at least one leaking well pipe.","The method comprises the use of granular particles of unconsolidated matter which, by means of their particle sorting, are arranged with a suitably small permeability, and wherein the particles of unconsolidated matter thereafter are mixed with water and potential other additives to become a fluidised mixture of unconsolidated matter (22) subsequently being placed, preferably by pumping, as a pressure- and flow-preventive barrier of unconsolidated matter (38, 56, 62, 68) in the pertinent annulus (16).","The barrier of unconsolidated matter (38, 56, 62, 68) is placed in the annulus (16) in such a way that inflowing fluids are brought into contact with, and are prevented from flowing by, the barrier of unconsolidated matter (38, 56, 62, 68) which, owing to the method, thus also is arranged with a suitably small permeability."],["1.A method for pressure- and flow-preventive fixing of at lease one size of well pipe (14, 54, 60, 66) in a well, including a petroleum well, during the drilling phase thereof, the method preventing fluids from flowing out of the well and also making cementation unnecessary for fixing said at least one size of well pipe (14, 54, 60, 66) in the well, wherein the method comprises: using granular particles of unconsolidated matter composed of a low-permeability-generating particle sorting; then mixing the particles of unconsolidated matter with at least water, possibly also other additives, including plasticizers, gelling agents and stabilisers, to form a fluidised mixture (22) thereof, characterised in that the method further comprises: placing said fluidised mixture (22) in at least a section of an annulus (16) positioned immediately external to said well pipe (14, 54, 60, 66), said annulus (16) being located between two casing sizes (14, 54, 60, 66) or, alternatively, between one casing size (14, 54, 60, 66) and a surrounding underground formation (12); and then allowing the particulate unconsolidated matter of the mixture (22) to set in said annulus (16), thereby fixing said at least one size of well pipe (14, 54, 60, 66) in the well, and thereby forming a pressure- and flow-preventive barrier (38, 56, 62, 68) of particulate unconsolidated matter therein preventing fluids from flowing out of the well.","2.A method according to claim 1, characterised in that the method further comprises: pumping said fluidised mixture (22) of particulate unconsolidated matter into said annulus (16), the mixture (22) being provided with properties allowing pumping thereof.","3.A method according to claim 2, characterised in that the method also comprises: concurrently pumping into said annulus (16) a cement slurry (26) immediately following said fluidised mixture (22) of particulate unconsolidated matter, said cement slurry (26), when set, forming a cement barrier (40, 58, 64, 70) placed between said barrier (38, 56, 62, 68) of unconsolidated matter and a bottom of the annulus (16), thereby preventing particles from the barrier (38, 56, 62, 68) of unconsolidated matter from falling down and into a potential new hole section (10) of the well.","4.Use of a fluidised mixture (22) of particulate unconsolidated matter, which is composed of a low-permeability-generating particle sorting, and a method according to one of the claims 1-3 for placing and setting a pressure- and flow-preventive barrier (38, 56, 62, 68) of said particulate unconsolidated matter in at least a section of an annulus (16) positioned immediately external to at least one size of well pipe (14, 54, 60, 66) in a well, including a petroleum well, during the drilling phase thereof, said barrier (38, 56, 62, 68) preventing fluids from, flowing out of the well and also making cementation unnecessary for fixing said at least one size of well pipe (14, 54, 60, 66) in the well."],["<SOH> BACKGROUND OF THE INVENTION <EOH>1.Field of the Invention The invention concerns a method for pressure- and flow-preventive fixing of pipes, for example casings and liners and possibly accompanying equipment, in a well when drilling the well.","The method may also be employed in a well, for example a completed well, in order to place one or several pressure- and flow-preventive barriers in one or several cavities/voids of the well, preferably annuli, wherein at least one adjoining pipe of the cavity/void/annulus is leaking.","The method according to the invention has developed mainly as is a consequence of a large and increasing need existing among authorities and industry, primarily the petroleum industry, to improve and eventually replace prior art methods for fixing casings in a well, prior art methods being encumbered with a series of severe problems and disadvantages, and cement being the primary prior art means for fixing casings and liners in the well.","2.Prior Art In connection with the drilling of a well, for example a petroleum well, and after having drilled a borehole down to a desired depth in the subsurface, it is customary to case the borehole with pipe(s).","Usually the well consists of several such boreholes, or hole sections, that sectionally and consecutively run with diminishing hole diameter into the subsurface.","It is therefore customary to provide the consecutive hole sections with casings of sectionally diminishing pipe diameters, wherein one casing size is placed within the preceding casing size etc.","Each casing size usually runs up to, and is connected to, the wellhead of the well.","So-called liners represent one exception to this which, on the other hand, do not run up to the wellhead of the well, and liners usually being employed to case one or several of the deepest hole sections of the well.","Such liners are usually fixedly cemented within and to a lower part of a preceding casing in such a way that the upper part of the liner overlaps the lower part of the preceding casing only.","Most casings, including liners, are fixed by cementing to the relevant borehole wall and usually also to the preceding casing.","In this context it is customary first to compute the amount of external annular volume of the pertinent casing to be filled with cement slurry, thereafter placing into said annulus/annuli a volume of cement slurry corresponding to at least that of the computed annular volume.","With the exception of liners, cementing of most casing sizes is carried out by pumping said volume of cement slurry down through the pertinent casing, thereafter forcing the cement slurry out/up into the annulus between the pertinent casing and the hole wall of the well and, eventually, usually also up into at least a lower part of the annulus between the pertinent casing and the preceding casing.","The cement slurry may be pumped in one or several stages, and into all or parts of the pertinent casing length, after which the cement slurry in principle shall harden into cement.","In the well, in order to avoid mixing, and thereby contaminating, the cement slurry with other liquids, usually drilling fluid, it is customary to place the cement slurry between two movable plugs, so-called wiper plugs, placed in the particular casing in order to facilitate the displacement of the cement slurry.","The lower and foremost of said plugs is a leading plug, while the upper and hindmost plug is a trailing plug.","Subsequently, and by means of pumping, the cement slurry and said plugs are displaced down through said casing.","The leading plug is arranged with a through-going hole which is covered by a diaphragm (a membrane), while the trailing plug usually is a solid and is substantially stronger than the leading plug.","By means of a fluid displacement column, usually a column of drilling fluid, placed on top of said trailing plug and arranged with necessary pumping equipment, the cement slurry and said plugs are subsequently pumped dorm through the casing until the leading plug is brought into contact with, and is arrested by, an associated seat or stopping device at the bottom of the casing.","Subsequently the pump pressure is sufficiently increased for said diaphragm to rupture, after which the cement slurry is pumped through said hole in the leading plug and is further displaced out/up into said annulus/annuli.","The pumping of cement slurry down through the casing continues until the trailing plug is brought into contact with, and is arrested by, the leading plug.","The displacement of cement slurry out/up into said annulus/annuli is thereby completed, but a sufficiently large liquid pressure is maintained in the overlying fluid displacement column for the cement slurry to harden without introducing movements in the cement slurry during the curing process.","In connection with fixedly cementing a liner in a well, however, a cementing pipe must be connected between cementing equipment at the surface of the well, for example at/on a drilling rig, and a lower part of said liner.","Usually, such a cementing pipe is comprised by a string consisting of connected drill pipes, the lower end part of the drill string being provided with an open and suitably adapted pipe, a so-called stinger, the stinger first being introduced into the well and being connected to a valve device located in the lower part of said liner.","Analogous to the above-described method, cement slurry and associated leading- and trailing plugs may subsequently be pumped down through the cementing pipe and onwards to said valve device, after which the cement slurry is displaced out/up into the external annulus of the liner.","In the hardened condition, the cement constitutes a fixed mass which, among other things, shall function as a pressure- and flow-preventive barrier in said annulus/annuli of the well.","In the event of potential fluid pressure differentials existing in the well, the cement shall prevent formation fluids from flowing between various formation layers and/or prevent formation fluids from flowing further upward in the well and possibly entirely to the surface.","Also, the cement shall maintain the casings fixed to the borehole wall of the well and usually also within and to a preceding casing.","For example, a surface casing of a well will largely support the weight of the other and smaller casing sizes of the well and also a wellhead or a blow-out preventer (“BOP”), and in this regard it is therefore necessary to establish a shear sustainable bond between the surface casing and the surrounding rocks, and in such a way that said loads may be transferred to the surrounding rocks.","Thus, the shear sustainable and load transferring bond often consists of cement.","Moreover, and upon commencing the drilling of the subsequent hole section, cement underlying and surrounding a casing shoe may contribute to stabilise a potentially fractured or unconsolidated rock in the hole wall of the well.","This stabilisation of said hole wall contributes to prevent or reduce the falling of rock fragments from the hole wall of said well region and into the subsequent hole section while the drilling thereof is carried out.","In order to drill a well down to a drilling objective, for example an oil/gas reservoir, usually it is of absolute necessity to place in the annulus of the well, a pressure- and flow-preventive mass, for example cement and/or a pressure- and flow-preventive device, potentially a sealing arrangement, for example a mechanical packer.","This particularly applies when drilling deep wells and/or when drilling wells down into subsurface layers wherein large fluid overpressures exist, simplistically denoted as overpressure.","An overpressure exists if the pores of a subsurface rock layer are exposed to a fluid pressure exceeding the liquid pressure which otherwise would exist if the layer was exposed to a normal hydrostatic pressure gradient from the surface and down to the subsurface layer of interest.","Upon drilling down through the various subsurface layers, a drilling fluid with a specific gravity, and thereby a hydrostatic liquid pressure, which is arranged to counteract the fluid pressure in the rock pores being penetrated, is used in the borehole.","This is done to prevent a potential and undesired inflow of formation fluids into the well.","When, during drilling at ambient conditions, a normal hydrostatic gradient exists in the subsurface pore fluids, normally the pressure gradient which may be observed in water-filled upper layers of the subsurface, said hydrostatic pore fluid pressure may be counteracted by arranging the drilling fluid with a slightly larger specific gravity/pressure gradient.","The various subsurface formation layers may also exhibit different properties of strength, wherein the rock strength largely may be related to lithological composition, particle distribution, particle cementation and degree of compaction of the subject rock.","Generally, the rock strength increases with increasing depth into the subsurface.","This implies that rocks being penetrated by a well, may be exposed to, and may resist, a gradually increasing fluid pressure without fracturing being initiated in the rocks.","A further increase of said fluid pressure will, however, result in fracturing of one or several of the penetrated rocks, this fracturing pressure commonly being denoted as the fracturing pressure of the subject rock(s), and the fracturing pressure commonly being recalculated, and expressed in terms of, an equivalent fracture gradient of the subject rock(s).","During drilling, upon approaching one or several formation layers with expected overpressures), the specific gravity/pressure gradient of the drilling fluid is increased to an extent necessary to withstand said overpressure(s).","Thus, potentially overpressured formation fluids are prevented from flowing into the well upon drilling into, potentially after having drilled into, said layer(s).","If said increase in the pressure gradient of the drilling fluid exceeds the fracture gradient of one or more of the penetrated rocks, the rocks(s) will be fractured and fractures develop in the rock(s).","Then, drilling fluid may flow unobstructedly out (leak) from the well and into the fractures, thereby causing the height of the drilling fluid column, and thus the liquid pressure in the liquid column, to be lowered.","By so doing, the formation pressure barrier brought about by the drilling fluid pressure exerted in the well is impaired, and this results in the establishment of an undesired, and potentially very dangerous, situation in the well.","In order to prevent such fracturing it is often absolutely necessary to isolate the penetrated formation layers from pressures that may fracture the rocks contained therein.","As mentioned, such a fracturing pressure may be exerted by the pressure, of the drilling fluid column, but the fracturing pressure may also be exerted by the overpressured formation fluids of other formation layers, usually deeper formation layers, which are being penetrated by the well during drilling.","Moreover, and pertaining to an open hole-section, it is the rock(s) of the shallowest part of the section, immediately underlying the casing shoe of the preceding casing, that generally, but not necessarily, is/are the weakest by strength, and thus being the one(s) which may first be fractured.","After having started the drilling of a new hole section in a well, it is for this reason common practise to undertake a so-called formation strength test of the shallowest rocks in said hole section.","Such a formation strength test is usually carried out immediately after having drilled the uppermost rocks along a 5-10 metre hole length of the new hole section.","For example, the formation strength test may consist in supplying said rocks with drilling fluid under a gradually increasing liquid pressure, and increasing the liquid pressure until an incipient fracturing of, and an accompanying leakage of drilling fluid into, the rocks is observed, which determines the fracturing pressure/fracture gradient of the rocks.","In the petroleum industry such a formation strength test is usually called a “leak-off test”.","In another commonly occurring formation strength test, a so-called formation integrity test, said rocks are also supplied with drilling fluid under a gradually increasing liquid pressure, limiting however the fluid pressure increase to a predefined maximum liquid pressure, and where this liquid pressure is considered to be the maximum required drilling fluid pressure to be applied for the new hole section to be drilled down to the desired drilling depths.","This maximum liquid pressure is usually smaller than the fracturing pressure of said rocks, thus not fracturing the rocks during this formation strength test.","Therefore, a formation integrity test is usually gentler on said rocks and the subsequent drilling operations than a fracturing test.","Such formation strength tests therefore provide a good indication as to the magnitude of liquid pressure, or magnitude of the liquid pressure gradient, whereby the drilling fluid may be arranged during the drilling of a hole section in order to avoid fracturing of the accompanying rocks.","Said maximum liquid pressure/liquid pressure gradient also limits the further drilling of a hole section to end at a depth at which the fluid pressure of a formation layer approaches said liquid pressure/liquid pressure gradient.","Cementing is also employed as a corrective method to prevent/reduce undesired inflow, and thereby also undesired pressure build-up, of a fluid in one or several regions of a well, including undesired fluid inflow through one or several leaking casings surrounding uncemented annuli of the well, the annulus/annuli possibly extending entirely up to the wellhead of the well.","The method consists in injecting cement slurry, possibly with the addition of plasticizing agents, gelling agents, stabilisers or other additives, into a relatively short annular interval covering said inflow region(s), whereupon the cement slurry or agent hardens or sets in such a way that it forms a pressure- and flow-preventive barrier which, in principle, shall prevent/reduce such fluid inflows."],["<SOH> BRIEF DESCRIPTION OF THE DRAWINGS <EOH>In the following part of the description, referring to FIGS.","1-5 , three non-limiting embodiments of the method according to the invention will be shown, one specific reference numeral referring to the same detail in all figures is where this detail is indicated, wherein: FIG.","1 and FIG.","2 show schematic vertical sections through a hole section of a well, in which hole section a casing is placed, and FIG.","1 shows a fluidised mixture of unconsolidated matter placed in said casing pending displacing of the mixture of unconsolidated matter out and up into an annulus surrounding the casing, while FIG.","2 shows the casing fixed in the hole section by means of the mixture of unconsolidated matter after it having been displaced out and up into said annulus, the mixture being placed as a pressure- and flow-preventive barrier of unconsolidated matter in the annulus; FIG.","3 and FIG.","4 also show schematic vertical sections through a segment of the hole section shown in FIG.","2 , the casing of the hole section being fixed in the well by means of the said barrier of unconsolidated matter in the surrounding annulus of the casing, and the figures show measures necessary in order to make a sidetrack of the well departing from said hole section, FIG.","3 showing perforation of the casing prior to a subsequent injection of cement slurry, while FIG.","4 shows a cut-through casing through which an introductory and new sidetrack hole section of the well is shown also; and FIG.","5 shows a schematic vertical section through several consecutive hole sections of a well, each hole section being provided with each their own casing size, and all casing sizes being fixed in the well by means of a barrier of unconsolidated matter placed in the surrounding annulus of each casing size.","detailed-description description=\"Detailed Description\" end=\"lead\"?"],["BACKGROUND OF THE INVENTION 1.Field of the Invention The invention concerns a method for pressure- and flow-preventive fixing of pipes, for example casings and liners and possibly accompanying equipment, in a well when drilling the well.","The method may also be employed in a well, for example a completed well, in order to place one or several pressure- and flow-preventive barriers in one or several cavities/voids of the well, preferably annuli, wherein at least one adjoining pipe of the cavity/void/annulus is leaking.","The method according to the invention has developed mainly as is a consequence of a large and increasing need existing among authorities and industry, primarily the petroleum industry, to improve and eventually replace prior art methods for fixing casings in a well, prior art methods being encumbered with a series of severe problems and disadvantages, and cement being the primary prior art means for fixing casings and liners in the well.","2.Prior Art In connection with the drilling of a well, for example a petroleum well, and after having drilled a borehole down to a desired depth in the subsurface, it is customary to case the borehole with pipe(s).","Usually the well consists of several such boreholes, or hole sections, that sectionally and consecutively run with diminishing hole diameter into the subsurface.","It is therefore customary to provide the consecutive hole sections with casings of sectionally diminishing pipe diameters, wherein one casing size is placed within the preceding casing size etc.","Each casing size usually runs up to, and is connected to, the wellhead of the well.","So-called liners represent one exception to this which, on the other hand, do not run up to the wellhead of the well, and liners usually being employed to case one or several of the deepest hole sections of the well.","Such liners are usually fixedly cemented within and to a lower part of a preceding casing in such a way that the upper part of the liner overlaps the lower part of the preceding casing only.","Most casings, including liners, are fixed by cementing to the relevant borehole wall and usually also to the preceding casing.","In this context it is customary first to compute the amount of external annular volume of the pertinent casing to be filled with cement slurry, thereafter placing into said annulus/annuli a volume of cement slurry corresponding to at least that of the computed annular volume.","With the exception of liners, cementing of most casing sizes is carried out by pumping said volume of cement slurry down through the pertinent casing, thereafter forcing the cement slurry out/up into the annulus between the pertinent casing and the hole wall of the well and, eventually, usually also up into at least a lower part of the annulus between the pertinent casing and the preceding casing.","The cement slurry may be pumped in one or several stages, and into all or parts of the pertinent casing length, after which the cement slurry in principle shall harden into cement.","In the well, in order to avoid mixing, and thereby contaminating, the cement slurry with other liquids, usually drilling fluid, it is customary to place the cement slurry between two movable plugs, so-called wiper plugs, placed in the particular casing in order to facilitate the displacement of the cement slurry.","The lower and foremost of said plugs is a leading plug, while the upper and hindmost plug is a trailing plug.","Subsequently, and by means of pumping, the cement slurry and said plugs are displaced down through said casing.","The leading plug is arranged with a through-going hole which is covered by a diaphragm (a membrane), while the trailing plug usually is a solid and is substantially stronger than the leading plug.","By means of a fluid displacement column, usually a column of drilling fluid, placed on top of said trailing plug and arranged with necessary pumping equipment, the cement slurry and said plugs are subsequently pumped dorm through the casing until the leading plug is brought into contact with, and is arrested by, an associated seat or stopping device at the bottom of the casing.","Subsequently the pump pressure is sufficiently increased for said diaphragm to rupture, after which the cement slurry is pumped through said hole in the leading plug and is further displaced out/up into said annulus/annuli.","The pumping of cement slurry down through the casing continues until the trailing plug is brought into contact with, and is arrested by, the leading plug.","The displacement of cement slurry out/up into said annulus/annuli is thereby completed, but a sufficiently large liquid pressure is maintained in the overlying fluid displacement column for the cement slurry to harden without introducing movements in the cement slurry during the curing process.","In connection with fixedly cementing a liner in a well, however, a cementing pipe must be connected between cementing equipment at the surface of the well, for example at/on a drilling rig, and a lower part of said liner.","Usually, such a cementing pipe is comprised by a string consisting of connected drill pipes, the lower end part of the drill string being provided with an open and suitably adapted pipe, a so-called stinger, the stinger first being introduced into the well and being connected to a valve device located in the lower part of said liner.","Analogous to the above-described method, cement slurry and associated leading- and trailing plugs may subsequently be pumped down through the cementing pipe and onwards to said valve device, after which the cement slurry is displaced out/up into the external annulus of the liner.","In the hardened condition, the cement constitutes a fixed mass which, among other things, shall function as a pressure- and flow-preventive barrier in said annulus/annuli of the well.","In the event of potential fluid pressure differentials existing in the well, the cement shall prevent formation fluids from flowing between various formation layers and/or prevent formation fluids from flowing further upward in the well and possibly entirely to the surface.","Also, the cement shall maintain the casings fixed to the borehole wall of the well and usually also within and to a preceding casing.","For example, a surface casing of a well will largely support the weight of the other and smaller casing sizes of the well and also a wellhead or a blow-out preventer (“BOP”), and in this regard it is therefore necessary to establish a shear sustainable bond between the surface casing and the surrounding rocks, and in such a way that said loads may be transferred to the surrounding rocks.","Thus, the shear sustainable and load transferring bond often consists of cement.","Moreover, and upon commencing the drilling of the subsequent hole section, cement underlying and surrounding a casing shoe may contribute to stabilise a potentially fractured or unconsolidated rock in the hole wall of the well.","This stabilisation of said hole wall contributes to prevent or reduce the falling of rock fragments from the hole wall of said well region and into the subsequent hole section while the drilling thereof is carried out.","In order to drill a well down to a drilling objective, for example an oil/gas reservoir, usually it is of absolute necessity to place in the annulus of the well, a pressure- and flow-preventive mass, for example cement and/or a pressure- and flow-preventive device, potentially a sealing arrangement, for example a mechanical packer.","This particularly applies when drilling deep wells and/or when drilling wells down into subsurface layers wherein large fluid overpressures exist, simplistically denoted as overpressure.","An overpressure exists if the pores of a subsurface rock layer are exposed to a fluid pressure exceeding the liquid pressure which otherwise would exist if the layer was exposed to a normal hydrostatic pressure gradient from the surface and down to the subsurface layer of interest.","Upon drilling down through the various subsurface layers, a drilling fluid with a specific gravity, and thereby a hydrostatic liquid pressure, which is arranged to counteract the fluid pressure in the rock pores being penetrated, is used in the borehole.","This is done to prevent a potential and undesired inflow of formation fluids into the well.","When, during drilling at ambient conditions, a normal hydrostatic gradient exists in the subsurface pore fluids, normally the pressure gradient which may be observed in water-filled upper layers of the subsurface, said hydrostatic pore fluid pressure may be counteracted by arranging the drilling fluid with a slightly larger specific gravity/pressure gradient.","The various subsurface formation layers may also exhibit different properties of strength, wherein the rock strength largely may be related to lithological composition, particle distribution, particle cementation and degree of compaction of the subject rock.","Generally, the rock strength increases with increasing depth into the subsurface.","This implies that rocks being penetrated by a well, may be exposed to, and may resist, a gradually increasing fluid pressure without fracturing being initiated in the rocks.","A further increase of said fluid pressure will, however, result in fracturing of one or several of the penetrated rocks, this fracturing pressure commonly being denoted as the fracturing pressure of the subject rock(s), and the fracturing pressure commonly being recalculated, and expressed in terms of, an equivalent fracture gradient of the subject rock(s).","During drilling, upon approaching one or several formation layers with expected overpressures), the specific gravity/pressure gradient of the drilling fluid is increased to an extent necessary to withstand said overpressure(s).","Thus, potentially overpressured formation fluids are prevented from flowing into the well upon drilling into, potentially after having drilled into, said layer(s).","If said increase in the pressure gradient of the drilling fluid exceeds the fracture gradient of one or more of the penetrated rocks, the rocks(s) will be fractured and fractures develop in the rock(s).","Then, drilling fluid may flow unobstructedly out (leak) from the well and into the fractures, thereby causing the height of the drilling fluid column, and thus the liquid pressure in the liquid column, to be lowered.","By so doing, the formation pressure barrier brought about by the drilling fluid pressure exerted in the well is impaired, and this results in the establishment of an undesired, and potentially very dangerous, situation in the well.","In order to prevent such fracturing it is often absolutely necessary to isolate the penetrated formation layers from pressures that may fracture the rocks contained therein.","As mentioned, such a fracturing pressure may be exerted by the pressure, of the drilling fluid column, but the fracturing pressure may also be exerted by the overpressured formation fluids of other formation layers, usually deeper formation layers, which are being penetrated by the well during drilling.","Moreover, and pertaining to an open hole-section, it is the rock(s) of the shallowest part of the section, immediately underlying the casing shoe of the preceding casing, that generally, but not necessarily, is/are the weakest by strength, and thus being the one(s) which may first be fractured.","After having started the drilling of a new hole section in a well, it is for this reason common practise to undertake a so-called formation strength test of the shallowest rocks in said hole section.","Such a formation strength test is usually carried out immediately after having drilled the uppermost rocks along a 5-10 metre hole length of the new hole section.","For example, the formation strength test may consist in supplying said rocks with drilling fluid under a gradually increasing liquid pressure, and increasing the liquid pressure until an incipient fracturing of, and an accompanying leakage of drilling fluid into, the rocks is observed, which determines the fracturing pressure/fracture gradient of the rocks.","In the petroleum industry such a formation strength test is usually called a “leak-off test”.","In another commonly occurring formation strength test, a so-called formation integrity test, said rocks are also supplied with drilling fluid under a gradually increasing liquid pressure, limiting however the fluid pressure increase to a predefined maximum liquid pressure, and where this liquid pressure is considered to be the maximum required drilling fluid pressure to be applied for the new hole section to be drilled down to the desired drilling depths.","This maximum liquid pressure is usually smaller than the fracturing pressure of said rocks, thus not fracturing the rocks during this formation strength test.","Therefore, a formation integrity test is usually gentler on said rocks and the subsequent drilling operations than a fracturing test.","Such formation strength tests therefore provide a good indication as to the magnitude of liquid pressure, or magnitude of the liquid pressure gradient, whereby the drilling fluid may be arranged during the drilling of a hole section in order to avoid fracturing of the accompanying rocks.","Said maximum liquid pressure/liquid pressure gradient also limits the further drilling of a hole section to end at a depth at which the fluid pressure of a formation layer approaches said liquid pressure/liquid pressure gradient.","Cementing is also employed as a corrective method to prevent/reduce undesired inflow, and thereby also undesired pressure build-up, of a fluid in one or several regions of a well, including undesired fluid inflow through one or several leaking casings surrounding uncemented annuli of the well, the annulus/annuli possibly extending entirely up to the wellhead of the well.","The method consists in injecting cement slurry, possibly with the addition of plasticizing agents, gelling agents, stabilisers or other additives, into a relatively short annular interval covering said inflow region(s), whereupon the cement slurry or agent hardens or sets in such a way that it forms a pressure- and flow-preventive barrier which, in principle, shall prevent/reduce such fluid inflows.","Disadvantages of the Prior Art Cementing jobs in a well are often encumbered with problems and disadvantages associated with the physical and chemical properties of the cement.","At the start of a cementing job, the cement exists in a liquid state as a slurry.","Later, and through a time-adapted curing process, the cement slurry is transformed to firm, or hardened, cement.","It is therefore of paramount importance that the cement slurry is placed into the intended cavity/void, usually an annulus, of the well while the cement slurry is sufficiently fluidised to enable it to displace onwards into this cavity/void/annulus.","Therefore, the placing of cement slurry into the well must be carried out before a significant thickening or hardening or the cement slurry has taken place.","If, during the placing into the well, the cement slurry is thickened or hardens prematurely, or if the cement slurry is introduced into and is thickened/hardened in an incorrect region/interval of the well, the cement will easily cause more problems than what it solves.","During the placing into the well, such a premature thickening/hardening of the cement slurry may develop if the slurry unintentionally is supplied with saline water, for example sea water or saline formation water.","Upon placing the slurry against a permeable formation layer of a surrounding formation hole wall, a premature thickening/hardening of the cement slurry may also develop in the event that the slurry water phase is being filtered and flows into said permeable formation layer.","If the cement slurry is thickened/hardened earlier than planned, thickened/hardened cement unintentionally may be placed in pipes and/or equipment otherwise intended to be open throughout.","For example, a premature thickening/hardening of the cement slurry in a cementing pipe and/or in a surrounding casing to be fixedly cemented, may result in the unintentional clogging of said pipes.","Correspondingly, a cement slurry being pumped down through, possibly in or around, (a) leaking pipe(s), may result in the unintentional and firm cementing of pipe(s) and/or equipment not to be fixedly cemented, hence resulting in said pipe(s) and/or equipment not working as intended, and possibly in not being able to remove the pipe/equipment from the well if or when this should become necessary.","For example, leakages in said cementation pipe and/or in the surrounding casing, may result in the cement slurry unintentionally being conducted onwards to the annulus between the outside of the cementing pipe and the surrounding casing, resulting in the cementing pipe unintentionally being fixed in said annulus upon thickening/hardening of the cement slurry.","In the worst case, such unintentional occurrences may result in having to re-drill all or parts of the well.","Said pipe leakages may also result in the cement slurry not displacing sufficiently far out/up into the relevant annulus to be fixedly cemented, which subsequently may result in the cement not exhibiting the desired pressure- and flow-preventive effect in the annulus.","In connection with such cementing jobs, channel-formed cavities/voids in the cement, so-called channelling in the cement, commonly develop, especially when cementing long pipe sections.","Such channelling in the cement represents an undesired effect that may result from the cement slurry and an accompanying liquid front between the cement slurry and an overlying drilling fluid, are being exposed, among other factors, to an uneven laminar flow while the slurry is being displaced out/up into an annulus in the well.","Such an uneven laminar flow often results in an insubstantially uniform and inefficient displacement of said liquid front in the annulus, drilling fluid channels thus being formed in the inflowing cement slurry as it flows out/up into the annulus, and resulting in said channels being maintained permanently in the annulus after hardening of the cement slurry.","Said annulus may be an annulus between two casings and/or an annulus between one casing and a surrounding formation hole wall.","Such channel-shaped cavities/voids in the cement often cause pressure- and fluid leakages.","Such pressure- and fluid leakages may also develop in connection with the curing process of the cement slurry.","initially, during the curing process, cement nuclei are formed, increasing gradually into a sufficiently large number to form a continuous lattice structure of cement nuclei, the lattice structure being sufficiently strong to carry the weight of newly formed cement nuclei.","At this stage of the curing process, when the load supporting lattice structure is established, and before consuming and chemically bonding the water phase of the cement slurry during the curing process, said water phase exists as an independent liquid in said lattice structure, the water phase at this stage of the curing process being exposed only to hydrostatic liquid pressures of its own and overlaying liquids.","However, the hydrostatic liquid pressure of the water phase is substantially less than the hydrostatic liquid pressure of the original cement slurry.","This reduction in hydrostatic liquid pressure may be sufficiently large for potential overpressured formation fluids of fluid-communicating formation layers to flow into the hardening slurry, causing subsequent pressure- and fluid leakages through it.","The presence of such formation fluids in the hardening cement slurry may prevent a further chemical reaction between cement and water in such a way that the function of the cement as a pressure- and flow-preventive barrier in the well, is impaired or destroyed.","It is obvious, however, that such impairments of the cement in a well may result in the overlying rocks thus not being sufficiently protected against pressure conditions that may cause fracturing of the rocks.","The fluid pressure from overpressured formation layers may thus propagate, via one or several annuli in the well, further upward in the well and, for example, cause an unintentional pressure build-up at the wellhead of the well.","At worst, such pressure- and fluid leakages may lead to an uncontrolled outflow of overpressured formation fluids at the surface of the well, a so-called surface blow-out; or to overpressured formation fluids flowing, via the well, between different formation layers, a so-called underground blow-out.","Furthermore, hardened cement, in the manner used in a well, constitutes a stiff, brittle and substantially inflexible material possessing a relatively large shear strength.","Advantageously, in some areas of utilisation, such material properties may be exploited.","For example, cement may be used as a load transferring connection between a surface casing and its surrounding formation hole wall.","As mentioned, in a borehole wall consisting of fractured or unconsolidated rocks, cement may be used as a shear-sustaining binding material binding together loose rocks and preventing the rocks from falling out from the hole wall and into the accompanying hole.","During the drilling of a hole section, such a falling out of loose rock fragments may cause large technical drilling problems if, for example, such unfastened rock fragments firmly pack around a drill string and prevent or stop any further drilling.","In other areas of utilisation, however, such material properties may appear less advantageous.","Some reservoirs consist, for example, of very porous sedimentary rocks, for example chalk or unconsolidated sand, such rocks often being soft and exhibiting very little material strength.","In deeper layers of the subsurface, such porous rocks are usually overpressured, which overpressure is and, through geologic time, has been a necessary prerequisite in order to preserve the porosity of a rock during its course of compaction.","In the process of recovering formation fluids from such a porous and weak reservoir rock, the formation pressure decreases gradually.","Consequently, a corresponding and gradual compression (compaction) of the rock pores also will take place, resulting in accompanying vertical movements in the reservoir rock and in the overlaying rocks.","However, well pipes, for example casings and/or liners, being placed in and throughout such compacting reservoir rocks are relatively.","rigid and not of a physical state such that they, as for the rock pores, may be pressed together, thereby compensating for vertical movements in the reservoir.","Consequently, relative movements take place between the well pipes and the surrounding rocks, and where the relative movements will tend to bend out/deflect, buckle/break and/or twist the pipes.","Moreover, and due to the stiffness, shear strength and compressive strength of the cement, cementing such well pipes to the surrounding rocks will further tend to prevent this bending/deflection, buckling and/or twisting.","Sufficiently large stress concentrations may thus be generated in the well pipes for one or several well pipes, in one or several places, to be torn to pieces or to be severely deformed.","Such a destruction or deformation of one or several well pipes may result in a production well becoming completely or partially abandoned, or in having to drill a new production well, thus incurring large technical and economical disadvantages.","The method of injecting cement slurry, possibly plasticizers, gelling agents, stabilisers or other additives into a relatively short annulus interval covering one or several undesired inflow area(s) in a well, is also encumbered with channels being formed and subsequent accompanying pressure- and fluid leakages in the cement.","Furthermore, production-related and relative pipe movements may also cause the cement to fracture or to unfasten from surrounding well pipes, and causing thus the cement to begin leaking.","Usually, such a cementing procedure will therefore only provide a usable pressure- and flow seal for a short period of time, after which problems of pressure build-up and potential fluid leakages may reappear in the well.","THE OBJECTIVES OF THE INVENTION One definite objective of this invention is to provide a new method for pressure- and flow-preventive fixing of pipes, for example casings and liners, and possibly accompanying equipment in a well.","Another definite objective of the invention is to be able to use the method in a completed well for the purpose of placing one or several pressure- and flow-preventive barriers in one or several annuli of which at least one pipe thereon is leaking.","The primary objective, however, is to be able to use the method, completely or partially, to replace the prior art functions of cement in a well, concurrently avoiding or reducing the above-mentioned problems and disadvantages associated with well cementing.","Achieving the Objectives Instead of placing cement slurry in the relevant cavity/void of the well, usually an annulus, the objectives are achieved by placing along a sufficient length of said cavity/void/annulus, a fluidised mixture of unconsolidated matter.","During the placing, the mixture of unconsolidated matter must be sufficiently fluidised for the mixture to displace onward to, and sufficiently far into, the cavity/void/annulus of interest.","In most applications, the placing may be carried out in the simplest and most efficient way by pumping the mixture of unconsolidated matter, as for cement slurry, through a connecting pipe onward to and into said cavity/void/annulus of the well.","In this respect, pipes and equipment of the types known in the art for fixedly cementing pipes in a well, may be used.","Largely, prior art methods for fixedly cementing such well pipes, may also be used for placing said mixture of unconsolidated matter into said cavity/void/annulus of the well.","Furthermore, and for the purpose of providing the fluidised mixture of unconsolidated matter with rheological properties enabling the mixture to be placed in the well, knowledge in the field of rheology together with devices, methods and additives which, for example, are used in the preparation and handling of drilling fluids/well cement, may be employed.","In order for such a mixture of unconsolidated matter to work as a pressure- and flow-preventive barrier in the well, the mixture of unconsolidated matter, being a substitute for cement, must be arranged in such a way in the well that it, when the fluidised unconsolidated matter has set in its operational position in the cavity/void/annulus, exhibits sufficiently good pressure- and flow-preventive properties.","In the method according to this invention, a mixture of unconsolidated matter comprised by naturally occurring and/or synthetically manufactured granular material, is therefore used in said barrier.","In the operation position in said cavity/void/annulus, the granular particles are assembled in such a way that they exhibit a very small permeability towards a fluid flowing through the mixture of unconsolidated matter.","Consequently, this method presupposes that the pressure- and flow-preventive barrier of unconsolidated matter is permeable, and that said fluid thereby leaks through the barrier of unconsolidated matter.","If the barrier of unconsolidated matter is arranged with a sufficiently small permeability over a sufficiently long length interval in the well, and a fluid is flowing through the barrier of unconsolidated matter, the fluid in the barrier of unconsolidated matter, however, will be exposed to a large flow resistance (flowing pressure drop) and thereby move very slowly (very small flow velocity) through the barrier of unconsolidated matter, and in such a way that the corresponding through-put flow time of the fluid theoretically may extend to several tens of thousands of years or more.","This course of flow is influenced by different parameters in accordance with Darcy's law which expresses a relation between several parameters and the flow velocity of a fluid when the fluid flows through a porous and permeable material; in which: v=k(Pin−Pout)/(μ·L); wherein ‘v’—flow velocity of the fluid (cm/sec), ‘k’—permeability of the material (Darcy), ‘Pin’—upstream pressure potential of the fluid (atmospheres), ‘Pout’—downstream pressure potential of the fluid (atmospheres), ‘(Pin−Pout)’—pressure loss through the material (atmospheres), ‘μ’—viscosity of the fluid (centipoise) ‘L’—length of permeable material (cm).","Considering that the fluid flow time through the barrier of unconsolidated matter theoretically is in the order of thousands of years, it is evident that the subsequent fluid leakage (amount of fluid leaking through the barrier) will be extremely small and, for practical purposes, negligible.","On the other hand, by using a cement barrier in a well, large pressure- and fluid leakages through the cement barrier often appear and are observed.","In the abovementioned perspective of time, however, such a cement barrier may constitute a substantially poorer, less enduring and insubstantially ductile/flexible barrier against pressure and flow than that of a barrier of unconsolidated matter.","The permeability ‘k’ of the mixture of unconsolidated matter and the extent or length ‘L’ of the barrier of unconsolidated matter in the well represent those parameters of Darcy's law which most readily may be influenced and controlled for the purpose of obtaining a sufficiently small fluid flow velocity ‘v’ through the barrier of unconsolidated matter.","Also, and to a lesser degree, the flow velocity ‘v’ may be influenced and controlled by selecting a suitable downstream pressure potential ‘Pout’ for the flowing fluid.","In practice, ‘Pout’ is comprised by the hydrostatic pressure being exerted on the barrier of unconsolidated matter by an overlaying/shallower liquid column, for example a water column, which hydrostatic pressure may be adapted, to some extent, by changing the specific gravity of the liquid column.","The upstream pressure potential ‘Pin’ of the fluid, however, usually is comprised by the formation pressure being exerted on the barrier of unconsolidated matter from an underlying/deeper reservoir layer, which pressure substantially may not be influenced/controlled, or may not be desired to be influenced/controlled, in consideration of said fluid flow velocity ‘v’ through the barrier of unconsolidated matter.","On the other hand, there may exist a desire to influence/control said formation pressure ‘Pin’ in consideration of the exploitation progress and degree of recovery of a reservoir, for example by implementing in the pertinent reservoir(s), actions of artificial stimulation, including water flooding.","According to Darcy's law, the permeability ‘k’ of the mixture of unconsolidated matter is proportional to the fluid flow velocity ‘v’ and, consequently, inversely proportional to the fluid through-put flow time, while the length ‘L’ of the barrier of unconsolidated matter is inversely proportional to the flow velocity ‘v’ and, consequently, proportional to the fluid through-put flow time.","By so doing, the fluid flow velocity ‘v’ and also through-put flow time may be controlled by selecting a suitable permeability ‘k’ and/or barrier length ‘L’.","In practice, considering that the maximum barrier length ‘L’ is limited to the length of the pertinent cavity/void/annulus of a well, the largest scope of influence/control on the flow velocity ‘v’/through-put flow time is gained by arranging the mixture of unconsolidated matter in such a way that it, in the operational position, exhibits a suitable permeability ‘k’.","The physical and chemical conditions prevailing in the subsurface layers of the individual well may vary from well to well.","Among other things, such physical and chemical conditions include reservoir depth, formation pressure(s) and temperature(s), type(s) of formation fluid(s) inclusive of its/their chemical compositions and physical properties, including properties or conditions influencing the viscosity of the fluid(s).","Considering that the prevailing physical and chemical conditions may vary from well to well, the permeability that is considered to be suitable for the pertinent well, also may vary from well to well.","Darcy's law shows, among other points, that the fluid viscosity ‘μ’ is inversely proportional to the fluid velocity ‘v’.","In one specific barrier of unconsolidated matter being arranged with a one specific permeability, a gas possessing a small viscosity, for example, will flow much faster through the barrier of unconsolidated matter than a heavy crude oil possessing a large viscosity is able to do in the same barrier.","In the event of wanting the gas and the crude oil to flow with equal flow velocity through each their own barrier of unconsolidated matter of equal (flow through-put) length, the barrier of unconsolidated matter for the gas must therefore be arranged with a substantially smaller permeability than that of the barrier of unconsolidated matter for the crude oil.","A barrier of unconsolidated matter should be arranged such that it, in the operational position, exhibits a permeability in the order of preferably, but not necessarily, a few milliDarcy (mD) and down to a level of microdarcy (μD), for example 0.001 mD (=1 μD).","In most wells, these are permeability values that will provide the desired pressure- and flow-preventive effect.","Nevertheless, for reasons mentioned above, the specific permeability of the barrier should be evaluated and determined based on the prevailing conditions in the pertinent well.","The mixture of unconsolidated matter is arranged with the desired permeability by it being composed of, and in the operational position consisting thereof, mixed granular particles of at least one particle size and, preferably, of several particle sizes.","Packed together, the permeability of the mixture of unconsolidated matter is determined by the geometric shape of a pore network comprised by the pores of the mixture of unconsolidated matter and their mutual pore connections.","The degree of variation in particle sizes has a large impact on the tightness at which the unconsolidated matter particles may be packed, which largely influences how the pore network will appear, hence also what the permeability of the mixture of unconsolidated matter will be.","Also, the general particle size of the mixture of unconsolidated matter are of great importance in determining how large the said pores and their mutual pore connections will be, which directly influences the permeability of the mixture of unconsolidated matter.","Consequently, one of two methods may be used to affect the permeability of such a mixture of unconsolidated matter; either by the mixture of unconsolidated matter being comprised of different particle sizes, or by the mixture of unconsolidated matter being comprised of small particles of a relatively homogenous size.","The distribution of particle sizes in such a mixture of unconsolidated matter is often expressed by the concept of sorting.","The concept of sorting is a qualitative measure of the degree of variation, or the range of variation, of different particle sizes in the mixture of unconsolidated matter.","A poorly sorted mixture of unconsolidated matter may include a large spectrum of particle sizes, for example particles in the size ranges of gravel, sand, silt and clay.","In comparison, a moderately sorted mixture of unconsolidated matter may include a small spectrum of particle sizes, for example medium sand and fine sand, while a very well sorted mass may include only one relatively homogenous particle size, for example coarse silt.","In the packed condition, such a poorly sorted mixture of unconsolidated matter may exhibit a very small permeability.","A very well sorted mixture of unconsolidated matter consisting of coarse silt may exhibit an equivalent small permeability, while a very well sorted mixture of unconsolidated matter consisting of very coarse sand may exhibit a very large permeability.","Such a specification of sorting, however, is imperfect for quantifying, or for specifying the quantities of, the different particle sizes comprising the mixture of unconsolidated matter.","On the other hand, the distribution of particle sizes in the mixture of unconsolidated matter may better be described and quantified by means of, for example, statistical concepts, wherein the distribution of particle sizes in the mixture of unconsolidated matter may be described by means of a cumulative distribution function.","In practice, different particle sizes may be provided by, for example, sieving and grouping naturally occurring granular unconsolidated matter into several different particle size categories.","Each such category is comprised of particles of a particular particle size range, the particle size range of each category differing from particle size ranges of possibly other categories.","Alternatively, synthetically manufactured granular material made of particle sizes within the pertinent particle size categories may be used.","Subsequently, certain amounts of particles of each of the pertinent particle size categories are assembled and mixed together, the mixture of unconsolidated matter thus being arranged with one particular distribution of particle sizes, hence one particular pore network shape of mixture of unconsolidated matter, which provides one particular permeability for the mixture of unconsolidated matter when placed in the operational position as a pressure- and flow-preventive barrier in the well.","Several scales exist specifying the different particle size categories, and the preferred scale may be related, largely, to particular trade disciplines.","The so-called Udden-Wentworth particle size scale and the so-called Krumbein phi (φ) particle size scale are generally known and are used, for one thing, in geologic disciplines, for example in sedimentology.","In the construction industry and in geotechnical environments, among other-matters, it is common, however, to use a scale referring to the mesh size (grate size) of a sieve device, for example the commonly used and so-called American Society of Testing and Materials (A.S.T.M.)","Sieve Scale.","The scale specifies particle size categories referring to so-called “mesh” sizes.","For example, a 200 mesh size represents 0.074 mm large sieve openings in an accompanying screen-cloth or grate of the sieve device.","Similar scales and/or concepts also exist which, to varying degrees, are used in different geographical regions and/or trade disciplines.","In the Udden-Wentworth scale, particles are grouped in particle size categories on the basis of average particle diameter specified in millimetres.","Examples of such size categories are fine gravel/granules (2-4 mm), very coarse sand (1-2 mm), coarse sand (0.5-1 mm), medium sand (0.25-0.5 mm), fine sand (0.125-0.25 mm), very fine sand (0.0625-0.125 mm), four categories of silt (0.0039-0.0625 mm), and also clay particles (<0.0039 mm).","In the Krumbein phi (φ) scale, the particle sizes are converted to φ-values, in which: φ=log2 d; wherein ‘d’—average particle diameter (mm).","Expressed in Krumbein φ-values, the preceding Udden-Wentworth examples of particle size categories may be specified as fine gravel/granules (φ=−2 to −1), very coarse sand (φ=−1 to 0), coarse sand (φ=0 to +1), medium sand (φ=+1 to +2), fine sand (φ=+2 to +3), very fine sand (φ=+3 to +4), four categories of silt (φ=+4 to +8), and also clay particles (φ=+8 or more).","Each particle size category being specified as integer φ-values, and not in fractions or decimal numerals, as in the Udden-Wentworth scale, such Krumbein φ-values are more easily treated statistically.","When using the Krumbein phi (φ) scale, the distribution of particle sizes in a mixture of unconsolidated matter (the sorting of the mixture of unconsolidated matter) is commonly specified as the range of variation (in φ-values) which includes an amount of particles comprising approximately ⅔ of all particles in the mixture of unconsolidated matter.","Statistically, this range of variation constitutes two times the standard deviation of the unconsolidated matter particles, and the standard deviation is therefore a commonly accepted measure of the sorting of a sediment or a mixture of unconsolidated matter.","In the construction industry and in geo-technical environments, among other things, it is customary to quantify a particular distribution of particle sizes (sorting) of a mixture of unconsolidated matter by means of a so-called sieve curve.","Specified in sieve- or mesh sizes, the sieve curve specifies the relative amounts, or the mass ratio, of the pertinent particle size categories which constitute, or is to constitute, the mixture of unconsolidated matter.","For example, the patent publication U.S. Pat.","No.","5,417,285 describes the use of a short plug of particulate material in connection with a partition or obstruction member in a well, the partition/obstruction member typically consisting of a mechanical plug, for example an inflatable packer or a so-called bridge plug.","Concurrently, and concerning the composition of the short particulate material plug, the publication describes seven different mixtures of particulate material and their specific and different particle compositions, each particle composition being expressed by means of “mesh” particle size categories and fractions by weight in percentages of the total weight of each mixture.","At the same time, the permeability of each mixture of particulate material has been determined by testing and specified in the publication.","Three of the said mixtures of particulate material showed especially a small permeability, and their particle compositions and permeabilities are of such a nature that they are considered suitable in a barrier of unconsolidated matter of the type comprised by this invention.","The particle composition and permeability of the three mixtures are stated in the following table summary: Test mixture - ref.","test-number cited in the publication Particle size 7 8 9 category Fraction by weight of the (mesh sizes) total weight of the mixture(%) 20/40 mesh sand 60 60 60 100 mesh sand 20 15 20 200 mesh sand 15 15 15 Bentonite “gel” 5 10 5 (clay fraction) Permeability (mD) 0.064 0.063 0.081 Said patent publication describes a 20/40 mesh sand as a conventional coarse sand, a 100 mesh sand as a conventional medium (“intermediate”) sand and a 200 mesh sand as a conventional fine sand, the test mixtures 7-9 also containing a particle fraction of fines described as a gel of bentonite/clay particles.","Chemically, the sand particles are described as consisting of preferably silica (silicon dioxide), mineralogically denoted as quartz.","Also, this is a suitable choice in that quartz (silicon dioxide) is one of the most weathering resistant minerals to be found in nature, and quartz/silica (silicon dioxide) therefore ought to provide excellent time- and weathering resistance in a well.","Moreover, the proprietors of the present invention have carried out laboratory experiments involving a similar mixture of unconsolidated matter.","Within a period of time of ca.","1.5 months, and by means of measurements, the permeability of the mixture of unconsolidated matter, among other things, was calculated as a function of the settling, or compaction, of the unconsolidated matter particles.","Also, the experiments confirmed that it is possible, in practice, to produce a fluidised mixture of unconsolidated matter which is easy to pump.","In the mixture of unconsolidated matter (predominantly silica/quartz), ca.","80 percent by weight of the mass consisted of sand size particles in the particle is size categories coarse sand (0.5-1 mm), medium sand (0.25-0.5 mm), fine sand (0.125-0.25 mm) and very fine-sand (0.0625-0.125 mm), while ca.","20 percent by weight of the mass consisted of silt size particles in the particle size range of 0.0039-0.0625 mm, half of which (ca.","10 percent by weight) in the size range of 0.005 mm (fine silt).","The last-mentioned fine silt particles were added to the mixture of unconsolidated matter exclusively to act as a permeability-reducing filler of the pores in the mixture since this fraction of mixture fines only contains negligible amounts of clay particles.","This differentiates this mixture of unconsolidated matter from the three mixtures specified in the preceding table, wherein relatively large fractions by weight of clay particles, so-called bentonite gel, are used in the mixtures, such suspended clay particles in the pores of the mixture acting as a binder between the particles.","Initially, a 1 metre length of mixture of unconsolidated matter was placed in the bottom of a vertically positioned plastic pipe, 6 metres long in total, after which the entire pipe was filled with fresh water.","During the subsequent time period of ca.","1.5 months, regular measurements were taken whereupon the permeability of the mixture of unconsolidated matter was calculated for the time period, observing during the time period diminishing permeability values.","At expiry of the time period, and after settling in the plastic pipe, the mixture could exhibit a permeability of 0.001 mD (=1 μD).","Furthermore, when being placed in the pipe, the mixture of unconsolidated matter was fluidised and contained ca.","83 percent by weight of unconsolidated matter particles and ca.","17 percent by weight of liquid, which consisted of ca.","11 percent by weight of water and ca.","6 percent by weight of a suitable plasticizer.","The plasticizer was used in order to avoid uneven settling of the fine-grained and coarse-grained particle fractions of the fluidised mixture of unconsolidated matter, but also in order to maintain the largest possible fraction of unconsolidated matter, hence the smallest possible fraction of liquid, in the fluidised mixture of unconsolidated matter.","Lignosulphonate represents one example of a common plasticizer/viscosity-regulating agent being used, for instance, in the petroleum industry, for example when preparing drilling fluids.","Moreover, these are merely examples of how such a mixture of unconsolidated matter may be composed, and of how the mixture of unconsolidated matter may be fluidised.","Further specifications of compositions of mixtures of unconsolidated matter, and also specifications of specific substances and agents, devices and methods known in the art for fluidising the mixture, are considered to be of prior art technical nature provided the method according to this invention is brought forward to a person skilled in art.","After having assembled and mixed in specific quantities of the pertinent particle size categories, for example as specified in the table mentioned above, and in such a way that the mixture of unconsolidated matter thus has been arranged with a distribution of particle sizes which, in the operational position, is to provide the desired permeability, the mixture of unconsolidated matter is fluidised prior to placing it in the relevant cavity/void/annulus of the well, this fluidisation simplifying the placing of the mixture of unconsolidated matter in the well.","For example, the fluidisation may be carried out by means of prior art devices and methods for stirring and mixing fluids and/or solids.","In this fluidisation process, the mixture of unconsolidated matter is mixed together with a suitable carrier fluid to become a fluidised mixture of unconsolidated matter, the fluidised mixture of unconsolidated matter being arranged in such a way that it thereafter, and preferably, may be pumped, for example by means of powerful cementing pumps and -equipment of the kind typically being used during the cementing of well pipes.","Most simplistically, the carrier fluid may be comprised of water.","On the other hand, plasticizers, gelling agents, stabilisers, weighting materials or other additives may be added in order to arrange the fluidised mixture of unconsolidated matter with appropriate physical and/or chemical properties, including rheological properties, to enable the mixture of unconsolidated matter to be placed and used as intended in the well.","For example, the fluidised mixture of unconsolidated matter must be arranged with a viscosity enabling the pumping of the mixture through, for example, said cementing pumps/-equipment and connected pipes in the well so that the fluidised mixture of unconsolidated matter may be displaced further out/up into the pertinent cavity/void/annulus of the well.","Also, and by example, the mixture may be arranged with suitable thixotropic properties.","Furthermore, the carrier fluid should constitute a minimal weight fraction, suitable for the purpose, of the mixture of unconsolidated matter.","Consequently, the fluidised mixture is comprised of a maximum weight fraction of unconsolidated matter particles forming the barrier of unconsolidated matter in the well, and this measure prevents or restricts a potential formation of surplus liquid originating from the carrier fluid.","During pumping, possibly after the pumping and in connection with the settling of the mixture of unconsolidated matter in said cavity/void/annulus, such measures advantageously result in avoiding or reducing premature settling (segregation) of possible coarse-grained fractions and, hence, in segregating these from remaining suspended finer-grained particle fractions in the fluidised mixture of unconsolidated matter.","A mixture of unconsolidated matter may thus be placed in the operational position in the well, a mixture which, after said settling, still is provided with the desired distribution and packing of particle sizes, thus also being provided with the desired permeability.","Such a potential uneven settling of particle sizes in the mixture of unconsolidated matter, as a barrier of unconsolidated matter in the well, may result in it exhibiting an uneven permeability distribution along its longitudinal extension in the well, and in the barrier of unconsolidated matter not exhibiting the desired pressure- and flow-preventive effect in the well.","Moreover, and in connection with the placing of the unconsolidated matter, care must be exercised in ensuring that the fluidised mixture of unconsolidated matter is arranged with a specific gravity not exceeding the fracturing pressure/fracture gradient of the relevant hole section of the well.","In this context, the fluidised and unconsolidated mixture possibly may be arranged with a specific gravity in the order of 2.1, which specific gravity does not differ from typical specific gravity values of a cement slurry.","Contrary to cement, such a barrier of unconsolidated matter may not harden in the cavity/void/annulus of the well.","In the operational position, the barrier of unconsolidated matter therefore may exhibit plastic properties, inasmuch as the barrier of unconsolidated matter is flexible and ductile and, simultaneously, possibly may exhibit insignificant tensile- and shear strength.","After having pressure- and flow-preventively fixed by means of the barrier of unconsolidated matter a specific casing size, these properties of such a barrier of unconsolidated matter must be considered in the event of commencing thereafter the drilling of a subsequent and deeper hole section.","If said casing size, entirely from the casing shoe and up throughout the well, is fixed by means of such a ductile mixture of unconsolidated matter, and due to its insignificant tensile- and shear strength, the unconsolidated matter may easily fall down and into the subsequent hole section when being drilled.","For instance, this problem may easily be avoided by placing in a length interval immediately underlying said barrier of unconsolidated matter, and in the same cavity/void/annulus, cement and/or another material possessing tensile- and shear strength, for example a mechanical annulus packer, and which prevents unconsolidated matter from falling down and into said hole section.","In practice, and in the very same well pipes, this may be carried out by pumping down into the well, and also out/up into said cavity/void/annulus, a volume of cement slurry concurrent with, and immediately following, the fluidised mixture of unconsolidated matter.","Afterwards, when the cement slurry is placed in its operational position and is hardening to form a cement barrier in the well, the mixture of unconsolidated matter will be prevented from falling down and into the subsequent hole section when being drilled.","This is shown in the following embodiments of the invention.","However, such a cement barrier need not be placed in the deepest casing/liner of the well, inasmuch as no subsequent hole section will be drilled into which the mixture of unconsolidated matter may fall down.","Advantags Achieved by the Invention Using one or several such barriers of unconsolidated matter in a well offers considerable advantages relative to the prior art, and relative especially to cementing of well pipes.","The placing of such a barrier of unconsolidated matter in a well does not involve, for example, a curing process that may create the above-described accompanying problems and disadvantages of cement, in which premature hardening/thickening of a cement slurry unintentionally may plug well pipes and -equipment, or possibly in unintentionally misplacing cement in the well.","Hence, the potential problems that may develop in a cement slurry while, during the curing process, forming a continuous lattice structure of cement nuclei therein, are avoided, the formation of this lattice structure possibly resulting eventually in potential overpressured formation fluids originating from fluid-communicating formation layers flowing into the hardening slurry and inflicting subsequent pressure- and fluid leakages through the resulting cement barrier, hence weakening or destroying a further chemical reaction between water and cement, which results in the pressure- and flow preventive function of the cement in the well being weakened or destroyed.","Such effects may not develop in a non-hardening barrier of unconsolidated matter because such a fluidised mixture of unconsolidated matter, during settling, will maintain its original liquid pressure/pressure gradient.","The absence of such a curing process, as well as said plastic properties of such a barrier of unconsolidated matter, also may result in the channelling in the fluidised mixture of unconsolidated matter not occurring, or occurring unsubstantially, when placed in the well.","Due to the ductility of the mixture of unconsolidated matter, potential channels being formed in the mixture of unconsolidated matter may be pressed together and disappear, completely or partially, during the ensuing settling period of the unconsolidated matter particles.","Potential fluids, for example drilling fluid, that have been trapped in such channels may thereby be displaced, completely or partially, out of the mixture of unconsolidated matter and not, as for a hardening cement slurry, inflict functional disturbance on the barrier in the well.","Also in posterity, awhile being placed as barrier of unconsolidated matter in a well, such a mixture of unconsolidated matter may retain its plastic properties and ductility.","Any movements and displacements that, during time, may take place in the surrounding rocks of the well, for example earth quake movements or movements in compacting reservoir rocks, may thus, as for a cement barrier, inflict onto a barrier of unconsolidated matter and accompanying well pipes stresses and accompanying relative movements.","As opposed to a cement barrier or a mechanical plug, the ductile barrier of unconsolidated matter, however, may be shaped according to, and adapt to, said relative movements without fractures and subsequent pressure- and fluid leakages being formed therein, and without substantially changing the small permeability of the barrier of unconsolidated matter.","Such movements potentially may cause the permeability of the barrier of unconsolidated matter to be further lowered, inasmuch as such influencing forces, in addition to the Earth's force of gravity, may contribute to packing more closely the particles of unconsolidated matter, and thus causing said permeability to be lowered.","Hence, the ductility and relative mobility of the barrier of unconsolidated matter cause well pipes to move in such a ductile mixture of unconsolidated matter, the well pipes being fixed by means of such a barrier of unconsolidated matter in the well wherein the well pipes are exposed to said stresses and relative movements.","By so doing, such well pipes may be exposed to a substantially larger relative movement in the form of bending, buckling and/or torsion than that or a cement barrier or a mechanical plug before one or several well pipes are torn to pieces or become severely deformed.","Advantageously, such a fluidised mixture of unconsolidated matter also may be injected into uncemented cavities/voids/annuli of a well being exposed to undesired inflow of fluids through one or several surrounding and leaking casings/liners.","The injection may be carried out, for example, through suitable perforations in a lower part of the casing/liner of the well, or via coiled tubing placed in an upper part of the pertinent annulus of the well.","The fluidised mixture of unconsolidated matter is placed in a suitable position in, and in a sufficient well length of, the pertinent cavity/annulus, for example in the entire length of the cavity/void/annulus, thus preventing/reducing pressure- and fluid leakages through said cavity/void/annulus, and without using, for example, cement and/or mechanical packers in the cavity/void/annulus.","By so doing, the lifetime of a leaking production tubing in a well may be extended in stead of having to re-complete or abandon the well.","Also, well pipes fixed by means of one or several such barriers of unconsolidated matter in a well provide for a significantly easier sidetracking or plugging, permanently or temporarily, of the well.","This is due to the unconsolidated matter of the barrier readily being removed in posterity, for example by flushing or circulating out the unconsolidated matter by means of a suitable liquid.","This substantially differs from the time-, equipment- and work consuming efforts being initiated in connection with the removal or drill-through of cement emplaced in connection with casing(s)/liner(s) of a well.","This is illustrated further in the following embodiments of the invention.","Relative to the prior art, including well cementing, the above-mentioned advantages show that the method according to the present invention provides a substantially cheaper technical solution which, moreover, is considerably simpler, more flexible and durable with time with respect to preventing/reducing pressure- and fluid leakages in a well, primarily in connection with the fixing of casing(s)/liner(s) of the well.","Also, the method may be used both in vertical, deviated and in horizontal wells.","BRIEF DESCRIPTION OF THE DRAWINGS In the following part of the description, referring to FIGS.","1-5, three non-limiting embodiments of the method according to the invention will be shown, one specific reference numeral referring to the same detail in all figures is where this detail is indicated, wherein: FIG.","1 and FIG.","2 show schematic vertical sections through a hole section of a well, in which hole section a casing is placed, and FIG.","1 shows a fluidised mixture of unconsolidated matter placed in said casing pending displacing of the mixture of unconsolidated matter out and up into an annulus surrounding the casing, while FIG.","2 shows the casing fixed in the hole section by means of the mixture of unconsolidated matter after it having been displaced out and up into said annulus, the mixture being placed as a pressure- and flow-preventive barrier of unconsolidated matter in the annulus; FIG.","3 and FIG.","4 also show schematic vertical sections through a segment of the hole section shown in FIG.","2, the casing of the hole section being fixed in the well by means of the said barrier of unconsolidated matter in the surrounding annulus of the casing, and the figures show measures necessary in order to make a sidetrack of the well departing from said hole section, FIG.","3 showing perforation of the casing prior to a subsequent injection of cement slurry, while FIG.","4 shows a cut-through casing through which an introductory and new sidetrack hole section of the well is shown also; and FIG.","5 shows a schematic vertical section through several consecutive hole sections of a well, each hole section being provided with each their own casing size, and all casing sizes being fixed in the well by means of a barrier of unconsolidated matter placed in the surrounding annulus of each casing size.","DESCRIPTION OF EMBODIMENTS OF THE INVENTION Knowledges, devices, appliances, equipment, agents, substances and/or methods known in the art not relating to the actual invention, but which nevertheless is/are, or may be, necessary prerequisites in order to practise the invention, will not be described in any detail in the following three embodiments.","Among other things, this includes pumping devices/-equipment and accompanying pipes being placed suitably in the well when practicing the invention.","Moreover, the figures only show details which are necessary in order to understand and practise the invention, and therefore the figures do not show, for example, a drilling arrangement and accompanying drilling equipment/well equipment etc.","The first embodiment is represented by FIG.","1 and FIG.","2, FIG.","1 showing a lower part of a hole section 10 in a subsurface well, the hole section 10 penetrating an underground formation 12.A casing 14 is placed in the hole section 10, and between the casing 14 and the hole section 10 an annulus 16 exists being filled with drilling fluid 18, the drilling fluid 18 also filling a volume at the bottom of the casing 14.Immediately overlying this volume are placed, in consecutive order, a first leading plug 20, a predefined volume of a fluidised mixture of unconsolidated matter 22 according to the previous description, a second leading plug 24, a predefined volume of cement slurry 26 and a trailing plug 28, the overlaying volume of the casing 14 being filled with drilling fluid 18.All plugs 20, 24 and 28 are placed in a pressure-sealing manner against the casing 14.Furthermore, the leading plugs 20 and 24 are arranged with each their own diaphragm 30 and 32 which, in connection with the subsequent pumping and an accompanying displacement of the mixture of unconsolidated matter 22, cement slurry 26 and the plugs 20, 24 and 28 down through the casing 14 and out/up into the annulus 16, are arranged to rupture when the diaphragms 30 and 32 are exposed to a sufficiently large pump pressure.","Furthermore, and associated with the diaphragms 30 and 32, each leading plug 20 and 24 is arranged with each their own through-going hole 34 and 36, through which the mixture of unconsolidated matter 22 and the cement slurry 26 may flow when the diaphragms 30 and 32 rupture due to said pump pressure.","These conditions have been further described in the preceding description.","However, the trailing plug 28 is massive and seats itself onto the second leading plug 24 at the end of the displacement, the second leading plug 24 being placed onto the first leading plug 20, and both leading plugs 20 and 24 being placed with ruptured diaphragms 30 and 32 in these positions.","FIG.","2 shows the plugs 20, 24 and 28 placed in these positions, and this figure also shows said mixture of unconsolidated matter 22 placed as a pressure- and flow-preventive barrier of unconsolidated matter 38 in the annulus 16, the unconsolidated matter being arranged with a suitable permeability by means of the distribution of particles described above, is arranged with a suitably small permeability, the annulus in a bottom interval of the well also being filled with an underlying hardened cement in the form of a cement barrier 40.The second embodiment is represented by FIG.","3 and FIG.","4.The embodiment describes measures necessary for sidetracking in the underground formation 12, and by means of prior art drilling equipment, a hole section 42 extending out from the well hole section 10 in the event of the casing 14 thereof being fixed in the annulus 16 by means of pressure- and flow-preventive particles of unconsolidated matter, cf.","the barrier of unconsolidated matter 38, FIG.","4 showing a segment of the sidetracked hole section 42.Moreover, this figure shows a hole 44 which, in order to enable sidetracking of the well, has been drilled through the casing 14 and the barrier of unconsolidated matter 38.The barrier of unconsolidated matter 38 surrounding the casing 14 exhibits insignificant tensile- and shear strength, and the drilling of a hole 44 through this barrier of unconsolidated matter 38 possibly may cause particles to unfasten and fall into the casing 14.By so doing, a section of length of the barrier or unconsolidated matter 38 being placed overlying the hole 44, may become destroyed, completely or partially, thus discontinuing, completely or partially, the function of the barrier of unconsolidated matter 38 as a pressure- and flow-preventive barrier.","This is, however, a problem which may be solved through simple means.","Prior to sidetracking, the casing 14 may be arranged with through-going perforations 46, cf.","FIG.","3, in an area of the hole section 10 overlying the region in which the drilling of the hole 44 for the ensuing sidetrack is desired.","Afterwards, a predefined volume of cement slurry is injected through the perforations 46 and into the barrier of unconsolidated matter 38.During the following hardening of the cement slurry, a cement plug 48 possessing substantial tensile- and shear strength is formed in the annulus 16.Thereafter, in a suitable position underlying the cement plug 48, the hole 44 through the casing 14 and the barrier of unconsolidated matter 38 may be drilled, inasmuch as the cement plug 48 prevents particles of unconsolidated matter from unfastening from an overlying part of the barrier of unconsolidated matter 38 and thereinafter falling down and into the casing 14.Then, the sidetracking of the hole section 42 may be carried out, the casing of the section 42 (casing not shown in the figures) possibly also being fixed by means of an equivalent barrier of unconsolidated matter.","If the last-mentioned casing is fixed by means of an equivalent barrier of unconsolidated matter, in which this barrier of unconsolidated matter also is to be placed in an associated annulus section overlying the hole 44, it is important to ensure that the corresponding mixture of unconsolidated matter, during the placing thereof in the annulus of the section 42, does not flow out through the hole 44 and down into the underlying pipe volume of the casing 14.Possibly, this problem may be solved by said pipe volume, prior to sidetracking, either being filled by a corresponding mixture of unconsolidated matter (not shown in the figures), or by placing, in a position immediately underlying the hole 44, a mechanical packer plug (not shown in the figure), possibly also by filling a corresponding mixture of unconsolidated matter between the mechanical packer plug and the hole 44, and in the casing 14.The third and last embodiment, represented by FIG.","5, shows a sidetracked well in the underground formation 12, the consecutive casing sizes of the well being pressure- and flow-preventively fixed by means of barriers of unconsolidated matter placed in respective and surrounding annuli thereof.","In this embodiment, the surface casing 50 of the well is fixed in the underground formation 12 by means of cement 52.Meanwhile, the subsequent first intermediate casing 54 of the well is fixed by means of a barrier of unconsolidated matter 56 and a short underlying cement barrier 58, and the subsequent second intermediate casing 60 is fixed by means of a barrier of unconsolidated matter 62 and a short underlying cement barrier 64, the barriers of unconsolidated matter 56 and 62 extending upward to, or near, the wellhead of the well (not shown in the figure).","The production casing 66 of the well is fixed by means of a barrier of unconsolidated matter 68 and a short underlying cement barrier 70, the barrier of unconsolidated matter 68 of this embodiment not extending upward to, or near, the wellhead of the well, but overlapping only a lower length interval of the preceding second intermediate casing 60.If the well is not drilled any deeper than that shown in FIG.","5, placing a cement barrier where the cement barrier 70 is shown in the figure is not required, strictly speaking, inasmuch as such a cement barrier 70 is placed in the well merely to prevent particles of unconsolidated matter from falling down and into a subsequent hole section (not shown in the figure)."]],"string":"[\n [\n \"Method for pressure-and flow-preventive fixing of pipes in a well\",\n \"A method for pressure- and flow-prevetive fixing of well pipes, preferably liners and casings (14, 54, 60, 66) in a well when drilling the well, wherein the method also may be used in a completed well in order to place a pressure- and flow-preventive barrier in an annulus (16) surrounded by at least one leaking well pipe.\",\n \"The method comprises the use of granular particles of unconsolidated matter which, by means of their particle sorting, are arranged with a suitably small permeability, and wherein the particles of unconsolidated matter thereafter are mixed with water and potential other additives to become a fluidised mixture of unconsolidated matter (22) subsequently being placed, preferably by pumping, as a pressure- and flow-preventive barrier of unconsolidated matter (38, 56, 62, 68) in the pertinent annulus (16).\",\n \"The barrier of unconsolidated matter (38, 56, 62, 68) is placed in the annulus (16) in such a way that inflowing fluids are brought into contact with, and are prevented from flowing by, the barrier of unconsolidated matter (38, 56, 62, 68) which, owing to the method, thus also is arranged with a suitably small permeability.\"\n ],\n [\n \"1.A method for pressure- and flow-preventive fixing of at lease one size of well pipe (14, 54, 60, 66) in a well, including a petroleum well, during the drilling phase thereof, the method preventing fluids from flowing out of the well and also making cementation unnecessary for fixing said at least one size of well pipe (14, 54, 60, 66) in the well, wherein the method comprises: using granular particles of unconsolidated matter composed of a low-permeability-generating particle sorting; then mixing the particles of unconsolidated matter with at least water, possibly also other additives, including plasticizers, gelling agents and stabilisers, to form a fluidised mixture (22) thereof, characterised in that the method further comprises: placing said fluidised mixture (22) in at least a section of an annulus (16) positioned immediately external to said well pipe (14, 54, 60, 66), said annulus (16) being located between two casing sizes (14, 54, 60, 66) or, alternatively, between one casing size (14, 54, 60, 66) and a surrounding underground formation (12); and then allowing the particulate unconsolidated matter of the mixture (22) to set in said annulus (16), thereby fixing said at least one size of well pipe (14, 54, 60, 66) in the well, and thereby forming a pressure- and flow-preventive barrier (38, 56, 62, 68) of particulate unconsolidated matter therein preventing fluids from flowing out of the well.\",\n \"2.A method according to claim 1, characterised in that the method further comprises: pumping said fluidised mixture (22) of particulate unconsolidated matter into said annulus (16), the mixture (22) being provided with properties allowing pumping thereof.\",\n \"3.A method according to claim 2, characterised in that the method also comprises: concurrently pumping into said annulus (16) a cement slurry (26) immediately following said fluidised mixture (22) of particulate unconsolidated matter, said cement slurry (26), when set, forming a cement barrier (40, 58, 64, 70) placed between said barrier (38, 56, 62, 68) of unconsolidated matter and a bottom of the annulus (16), thereby preventing particles from the barrier (38, 56, 62, 68) of unconsolidated matter from falling down and into a potential new hole section (10) of the well.\",\n \"4.Use of a fluidised mixture (22) of particulate unconsolidated matter, which is composed of a low-permeability-generating particle sorting, and a method according to one of the claims 1-3 for placing and setting a pressure- and flow-preventive barrier (38, 56, 62, 68) of said particulate unconsolidated matter in at least a section of an annulus (16) positioned immediately external to at least one size of well pipe (14, 54, 60, 66) in a well, including a petroleum well, during the drilling phase thereof, said barrier (38, 56, 62, 68) preventing fluids from, flowing out of the well and also making cementation unnecessary for fixing said at least one size of well pipe (14, 54, 60, 66) in the well.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>1.Field of the Invention The invention concerns a method for pressure- and flow-preventive fixing of pipes, for example casings and liners and possibly accompanying equipment, in a well when drilling the well.\",\n \"The method may also be employed in a well, for example a completed well, in order to place one or several pressure- and flow-preventive barriers in one or several cavities/voids of the well, preferably annuli, wherein at least one adjoining pipe of the cavity/void/annulus is leaking.\",\n \"The method according to the invention has developed mainly as is a consequence of a large and increasing need existing among authorities and industry, primarily the petroleum industry, to improve and eventually replace prior art methods for fixing casings in a well, prior art methods being encumbered with a series of severe problems and disadvantages, and cement being the primary prior art means for fixing casings and liners in the well.\",\n \"2.Prior Art In connection with the drilling of a well, for example a petroleum well, and after having drilled a borehole down to a desired depth in the subsurface, it is customary to case the borehole with pipe(s).\",\n \"Usually the well consists of several such boreholes, or hole sections, that sectionally and consecutively run with diminishing hole diameter into the subsurface.\",\n \"It is therefore customary to provide the consecutive hole sections with casings of sectionally diminishing pipe diameters, wherein one casing size is placed within the preceding casing size etc.\",\n \"Each casing size usually runs up to, and is connected to, the wellhead of the well.\",\n \"So-called liners represent one exception to this which, on the other hand, do not run up to the wellhead of the well, and liners usually being employed to case one or several of the deepest hole sections of the well.\",\n \"Such liners are usually fixedly cemented within and to a lower part of a preceding casing in such a way that the upper part of the liner overlaps the lower part of the preceding casing only.\",\n \"Most casings, including liners, are fixed by cementing to the relevant borehole wall and usually also to the preceding casing.\",\n \"In this context it is customary first to compute the amount of external annular volume of the pertinent casing to be filled with cement slurry, thereafter placing into said annulus/annuli a volume of cement slurry corresponding to at least that of the computed annular volume.\",\n \"With the exception of liners, cementing of most casing sizes is carried out by pumping said volume of cement slurry down through the pertinent casing, thereafter forcing the cement slurry out/up into the annulus between the pertinent casing and the hole wall of the well and, eventually, usually also up into at least a lower part of the annulus between the pertinent casing and the preceding casing.\",\n \"The cement slurry may be pumped in one or several stages, and into all or parts of the pertinent casing length, after which the cement slurry in principle shall harden into cement.\",\n \"In the well, in order to avoid mixing, and thereby contaminating, the cement slurry with other liquids, usually drilling fluid, it is customary to place the cement slurry between two movable plugs, so-called wiper plugs, placed in the particular casing in order to facilitate the displacement of the cement slurry.\",\n \"The lower and foremost of said plugs is a leading plug, while the upper and hindmost plug is a trailing plug.\",\n \"Subsequently, and by means of pumping, the cement slurry and said plugs are displaced down through said casing.\",\n \"The leading plug is arranged with a through-going hole which is covered by a diaphragm (a membrane), while the trailing plug usually is a solid and is substantially stronger than the leading plug.\",\n \"By means of a fluid displacement column, usually a column of drilling fluid, placed on top of said trailing plug and arranged with necessary pumping equipment, the cement slurry and said plugs are subsequently pumped dorm through the casing until the leading plug is brought into contact with, and is arrested by, an associated seat or stopping device at the bottom of the casing.\",\n \"Subsequently the pump pressure is sufficiently increased for said diaphragm to rupture, after which the cement slurry is pumped through said hole in the leading plug and is further displaced out/up into said annulus/annuli.\",\n \"The pumping of cement slurry down through the casing continues until the trailing plug is brought into contact with, and is arrested by, the leading plug.\",\n \"The displacement of cement slurry out/up into said annulus/annuli is thereby completed, but a sufficiently large liquid pressure is maintained in the overlying fluid displacement column for the cement slurry to harden without introducing movements in the cement slurry during the curing process.\",\n \"In connection with fixedly cementing a liner in a well, however, a cementing pipe must be connected between cementing equipment at the surface of the well, for example at/on a drilling rig, and a lower part of said liner.\",\n \"Usually, such a cementing pipe is comprised by a string consisting of connected drill pipes, the lower end part of the drill string being provided with an open and suitably adapted pipe, a so-called stinger, the stinger first being introduced into the well and being connected to a valve device located in the lower part of said liner.\",\n \"Analogous to the above-described method, cement slurry and associated leading- and trailing plugs may subsequently be pumped down through the cementing pipe and onwards to said valve device, after which the cement slurry is displaced out/up into the external annulus of the liner.\",\n \"In the hardened condition, the cement constitutes a fixed mass which, among other things, shall function as a pressure- and flow-preventive barrier in said annulus/annuli of the well.\",\n \"In the event of potential fluid pressure differentials existing in the well, the cement shall prevent formation fluids from flowing between various formation layers and/or prevent formation fluids from flowing further upward in the well and possibly entirely to the surface.\",\n \"Also, the cement shall maintain the casings fixed to the borehole wall of the well and usually also within and to a preceding casing.\",\n \"For example, a surface casing of a well will largely support the weight of the other and smaller casing sizes of the well and also a wellhead or a blow-out preventer (“BOP”), and in this regard it is therefore necessary to establish a shear sustainable bond between the surface casing and the surrounding rocks, and in such a way that said loads may be transferred to the surrounding rocks.\",\n \"Thus, the shear sustainable and load transferring bond often consists of cement.\",\n \"Moreover, and upon commencing the drilling of the subsequent hole section, cement underlying and surrounding a casing shoe may contribute to stabilise a potentially fractured or unconsolidated rock in the hole wall of the well.\",\n \"This stabilisation of said hole wall contributes to prevent or reduce the falling of rock fragments from the hole wall of said well region and into the subsequent hole section while the drilling thereof is carried out.\",\n \"In order to drill a well down to a drilling objective, for example an oil/gas reservoir, usually it is of absolute necessity to place in the annulus of the well, a pressure- and flow-preventive mass, for example cement and/or a pressure- and flow-preventive device, potentially a sealing arrangement, for example a mechanical packer.\",\n \"This particularly applies when drilling deep wells and/or when drilling wells down into subsurface layers wherein large fluid overpressures exist, simplistically denoted as overpressure.\",\n \"An overpressure exists if the pores of a subsurface rock layer are exposed to a fluid pressure exceeding the liquid pressure which otherwise would exist if the layer was exposed to a normal hydrostatic pressure gradient from the surface and down to the subsurface layer of interest.\",\n \"Upon drilling down through the various subsurface layers, a drilling fluid with a specific gravity, and thereby a hydrostatic liquid pressure, which is arranged to counteract the fluid pressure in the rock pores being penetrated, is used in the borehole.\",\n \"This is done to prevent a potential and undesired inflow of formation fluids into the well.\",\n \"When, during drilling at ambient conditions, a normal hydrostatic gradient exists in the subsurface pore fluids, normally the pressure gradient which may be observed in water-filled upper layers of the subsurface, said hydrostatic pore fluid pressure may be counteracted by arranging the drilling fluid with a slightly larger specific gravity/pressure gradient.\",\n \"The various subsurface formation layers may also exhibit different properties of strength, wherein the rock strength largely may be related to lithological composition, particle distribution, particle cementation and degree of compaction of the subject rock.\",\n \"Generally, the rock strength increases with increasing depth into the subsurface.\",\n \"This implies that rocks being penetrated by a well, may be exposed to, and may resist, a gradually increasing fluid pressure without fracturing being initiated in the rocks.\",\n \"A further increase of said fluid pressure will, however, result in fracturing of one or several of the penetrated rocks, this fracturing pressure commonly being denoted as the fracturing pressure of the subject rock(s), and the fracturing pressure commonly being recalculated, and expressed in terms of, an equivalent fracture gradient of the subject rock(s).\",\n \"During drilling, upon approaching one or several formation layers with expected overpressures), the specific gravity/pressure gradient of the drilling fluid is increased to an extent necessary to withstand said overpressure(s).\",\n \"Thus, potentially overpressured formation fluids are prevented from flowing into the well upon drilling into, potentially after having drilled into, said layer(s).\",\n \"If said increase in the pressure gradient of the drilling fluid exceeds the fracture gradient of one or more of the penetrated rocks, the rocks(s) will be fractured and fractures develop in the rock(s).\",\n \"Then, drilling fluid may flow unobstructedly out (leak) from the well and into the fractures, thereby causing the height of the drilling fluid column, and thus the liquid pressure in the liquid column, to be lowered.\",\n \"By so doing, the formation pressure barrier brought about by the drilling fluid pressure exerted in the well is impaired, and this results in the establishment of an undesired, and potentially very dangerous, situation in the well.\",\n \"In order to prevent such fracturing it is often absolutely necessary to isolate the penetrated formation layers from pressures that may fracture the rocks contained therein.\",\n \"As mentioned, such a fracturing pressure may be exerted by the pressure, of the drilling fluid column, but the fracturing pressure may also be exerted by the overpressured formation fluids of other formation layers, usually deeper formation layers, which are being penetrated by the well during drilling.\",\n \"Moreover, and pertaining to an open hole-section, it is the rock(s) of the shallowest part of the section, immediately underlying the casing shoe of the preceding casing, that generally, but not necessarily, is/are the weakest by strength, and thus being the one(s) which may first be fractured.\",\n \"After having started the drilling of a new hole section in a well, it is for this reason common practise to undertake a so-called formation strength test of the shallowest rocks in said hole section.\",\n \"Such a formation strength test is usually carried out immediately after having drilled the uppermost rocks along a 5-10 metre hole length of the new hole section.\",\n \"For example, the formation strength test may consist in supplying said rocks with drilling fluid under a gradually increasing liquid pressure, and increasing the liquid pressure until an incipient fracturing of, and an accompanying leakage of drilling fluid into, the rocks is observed, which determines the fracturing pressure/fracture gradient of the rocks.\",\n \"In the petroleum industry such a formation strength test is usually called a “leak-off test”.\",\n \"In another commonly occurring formation strength test, a so-called formation integrity test, said rocks are also supplied with drilling fluid under a gradually increasing liquid pressure, limiting however the fluid pressure increase to a predefined maximum liquid pressure, and where this liquid pressure is considered to be the maximum required drilling fluid pressure to be applied for the new hole section to be drilled down to the desired drilling depths.\",\n \"This maximum liquid pressure is usually smaller than the fracturing pressure of said rocks, thus not fracturing the rocks during this formation strength test.\",\n \"Therefore, a formation integrity test is usually gentler on said rocks and the subsequent drilling operations than a fracturing test.\",\n \"Such formation strength tests therefore provide a good indication as to the magnitude of liquid pressure, or magnitude of the liquid pressure gradient, whereby the drilling fluid may be arranged during the drilling of a hole section in order to avoid fracturing of the accompanying rocks.\",\n \"Said maximum liquid pressure/liquid pressure gradient also limits the further drilling of a hole section to end at a depth at which the fluid pressure of a formation layer approaches said liquid pressure/liquid pressure gradient.\",\n \"Cementing is also employed as a corrective method to prevent/reduce undesired inflow, and thereby also undesired pressure build-up, of a fluid in one or several regions of a well, including undesired fluid inflow through one or several leaking casings surrounding uncemented annuli of the well, the annulus/annuli possibly extending entirely up to the wellhead of the well.\",\n \"The method consists in injecting cement slurry, possibly with the addition of plasticizing agents, gelling agents, stabilisers or other additives, into a relatively short annular interval covering said inflow region(s), whereupon the cement slurry or agent hardens or sets in such a way that it forms a pressure- and flow-preventive barrier which, in principle, shall prevent/reduce such fluid inflows.\"\n ],\n [\n \"<SOH> BRIEF DESCRIPTION OF THE DRAWINGS <EOH>In the following part of the description, referring to FIGS.\",\n \"1-5 , three non-limiting embodiments of the method according to the invention will be shown, one specific reference numeral referring to the same detail in all figures is where this detail is indicated, wherein: FIG.\",\n \"1 and FIG.\",\n \"2 show schematic vertical sections through a hole section of a well, in which hole section a casing is placed, and FIG.\",\n \"1 shows a fluidised mixture of unconsolidated matter placed in said casing pending displacing of the mixture of unconsolidated matter out and up into an annulus surrounding the casing, while FIG.\",\n \"2 shows the casing fixed in the hole section by means of the mixture of unconsolidated matter after it having been displaced out and up into said annulus, the mixture being placed as a pressure- and flow-preventive barrier of unconsolidated matter in the annulus; FIG.\",\n \"3 and FIG.\",\n \"4 also show schematic vertical sections through a segment of the hole section shown in FIG.\",\n \"2 , the casing of the hole section being fixed in the well by means of the said barrier of unconsolidated matter in the surrounding annulus of the casing, and the figures show measures necessary in order to make a sidetrack of the well departing from said hole section, FIG.\",\n \"3 showing perforation of the casing prior to a subsequent injection of cement slurry, while FIG.\",\n \"4 shows a cut-through casing through which an introductory and new sidetrack hole section of the well is shown also; and FIG.\",\n \"5 shows a schematic vertical section through several consecutive hole sections of a well, each hole section being provided with each their own casing size, and all casing sizes being fixed in the well by means of a barrier of unconsolidated matter placed in the surrounding annulus of each casing size.\",\n \"detailed-description description=\\\"Detailed Description\\\" end=\\\"lead\\\"?\"\n ],\n [\n \"BACKGROUND OF THE INVENTION 1.Field of the Invention The invention concerns a method for pressure- and flow-preventive fixing of pipes, for example casings and liners and possibly accompanying equipment, in a well when drilling the well.\",\n \"The method may also be employed in a well, for example a completed well, in order to place one or several pressure- and flow-preventive barriers in one or several cavities/voids of the well, preferably annuli, wherein at least one adjoining pipe of the cavity/void/annulus is leaking.\",\n \"The method according to the invention has developed mainly as is a consequence of a large and increasing need existing among authorities and industry, primarily the petroleum industry, to improve and eventually replace prior art methods for fixing casings in a well, prior art methods being encumbered with a series of severe problems and disadvantages, and cement being the primary prior art means for fixing casings and liners in the well.\",\n \"2.Prior Art In connection with the drilling of a well, for example a petroleum well, and after having drilled a borehole down to a desired depth in the subsurface, it is customary to case the borehole with pipe(s).\",\n \"Usually the well consists of several such boreholes, or hole sections, that sectionally and consecutively run with diminishing hole diameter into the subsurface.\",\n \"It is therefore customary to provide the consecutive hole sections with casings of sectionally diminishing pipe diameters, wherein one casing size is placed within the preceding casing size etc.\",\n \"Each casing size usually runs up to, and is connected to, the wellhead of the well.\",\n \"So-called liners represent one exception to this which, on the other hand, do not run up to the wellhead of the well, and liners usually being employed to case one or several of the deepest hole sections of the well.\",\n \"Such liners are usually fixedly cemented within and to a lower part of a preceding casing in such a way that the upper part of the liner overlaps the lower part of the preceding casing only.\",\n \"Most casings, including liners, are fixed by cementing to the relevant borehole wall and usually also to the preceding casing.\",\n \"In this context it is customary first to compute the amount of external annular volume of the pertinent casing to be filled with cement slurry, thereafter placing into said annulus/annuli a volume of cement slurry corresponding to at least that of the computed annular volume.\",\n \"With the exception of liners, cementing of most casing sizes is carried out by pumping said volume of cement slurry down through the pertinent casing, thereafter forcing the cement slurry out/up into the annulus between the pertinent casing and the hole wall of the well and, eventually, usually also up into at least a lower part of the annulus between the pertinent casing and the preceding casing.\",\n \"The cement slurry may be pumped in one or several stages, and into all or parts of the pertinent casing length, after which the cement slurry in principle shall harden into cement.\",\n \"In the well, in order to avoid mixing, and thereby contaminating, the cement slurry with other liquids, usually drilling fluid, it is customary to place the cement slurry between two movable plugs, so-called wiper plugs, placed in the particular casing in order to facilitate the displacement of the cement slurry.\",\n \"The lower and foremost of said plugs is a leading plug, while the upper and hindmost plug is a trailing plug.\",\n \"Subsequently, and by means of pumping, the cement slurry and said plugs are displaced down through said casing.\",\n \"The leading plug is arranged with a through-going hole which is covered by a diaphragm (a membrane), while the trailing plug usually is a solid and is substantially stronger than the leading plug.\",\n \"By means of a fluid displacement column, usually a column of drilling fluid, placed on top of said trailing plug and arranged with necessary pumping equipment, the cement slurry and said plugs are subsequently pumped dorm through the casing until the leading plug is brought into contact with, and is arrested by, an associated seat or stopping device at the bottom of the casing.\",\n \"Subsequently the pump pressure is sufficiently increased for said diaphragm to rupture, after which the cement slurry is pumped through said hole in the leading plug and is further displaced out/up into said annulus/annuli.\",\n \"The pumping of cement slurry down through the casing continues until the trailing plug is brought into contact with, and is arrested by, the leading plug.\",\n \"The displacement of cement slurry out/up into said annulus/annuli is thereby completed, but a sufficiently large liquid pressure is maintained in the overlying fluid displacement column for the cement slurry to harden without introducing movements in the cement slurry during the curing process.\",\n \"In connection with fixedly cementing a liner in a well, however, a cementing pipe must be connected between cementing equipment at the surface of the well, for example at/on a drilling rig, and a lower part of said liner.\",\n \"Usually, such a cementing pipe is comprised by a string consisting of connected drill pipes, the lower end part of the drill string being provided with an open and suitably adapted pipe, a so-called stinger, the stinger first being introduced into the well and being connected to a valve device located in the lower part of said liner.\",\n \"Analogous to the above-described method, cement slurry and associated leading- and trailing plugs may subsequently be pumped down through the cementing pipe and onwards to said valve device, after which the cement slurry is displaced out/up into the external annulus of the liner.\",\n \"In the hardened condition, the cement constitutes a fixed mass which, among other things, shall function as a pressure- and flow-preventive barrier in said annulus/annuli of the well.\",\n \"In the event of potential fluid pressure differentials existing in the well, the cement shall prevent formation fluids from flowing between various formation layers and/or prevent formation fluids from flowing further upward in the well and possibly entirely to the surface.\",\n \"Also, the cement shall maintain the casings fixed to the borehole wall of the well and usually also within and to a preceding casing.\",\n \"For example, a surface casing of a well will largely support the weight of the other and smaller casing sizes of the well and also a wellhead or a blow-out preventer (“BOP”), and in this regard it is therefore necessary to establish a shear sustainable bond between the surface casing and the surrounding rocks, and in such a way that said loads may be transferred to the surrounding rocks.\",\n \"Thus, the shear sustainable and load transferring bond often consists of cement.\",\n \"Moreover, and upon commencing the drilling of the subsequent hole section, cement underlying and surrounding a casing shoe may contribute to stabilise a potentially fractured or unconsolidated rock in the hole wall of the well.\",\n \"This stabilisation of said hole wall contributes to prevent or reduce the falling of rock fragments from the hole wall of said well region and into the subsequent hole section while the drilling thereof is carried out.\",\n \"In order to drill a well down to a drilling objective, for example an oil/gas reservoir, usually it is of absolute necessity to place in the annulus of the well, a pressure- and flow-preventive mass, for example cement and/or a pressure- and flow-preventive device, potentially a sealing arrangement, for example a mechanical packer.\",\n \"This particularly applies when drilling deep wells and/or when drilling wells down into subsurface layers wherein large fluid overpressures exist, simplistically denoted as overpressure.\",\n \"An overpressure exists if the pores of a subsurface rock layer are exposed to a fluid pressure exceeding the liquid pressure which otherwise would exist if the layer was exposed to a normal hydrostatic pressure gradient from the surface and down to the subsurface layer of interest.\",\n \"Upon drilling down through the various subsurface layers, a drilling fluid with a specific gravity, and thereby a hydrostatic liquid pressure, which is arranged to counteract the fluid pressure in the rock pores being penetrated, is used in the borehole.\",\n \"This is done to prevent a potential and undesired inflow of formation fluids into the well.\",\n \"When, during drilling at ambient conditions, a normal hydrostatic gradient exists in the subsurface pore fluids, normally the pressure gradient which may be observed in water-filled upper layers of the subsurface, said hydrostatic pore fluid pressure may be counteracted by arranging the drilling fluid with a slightly larger specific gravity/pressure gradient.\",\n \"The various subsurface formation layers may also exhibit different properties of strength, wherein the rock strength largely may be related to lithological composition, particle distribution, particle cementation and degree of compaction of the subject rock.\",\n \"Generally, the rock strength increases with increasing depth into the subsurface.\",\n \"This implies that rocks being penetrated by a well, may be exposed to, and may resist, a gradually increasing fluid pressure without fracturing being initiated in the rocks.\",\n \"A further increase of said fluid pressure will, however, result in fracturing of one or several of the penetrated rocks, this fracturing pressure commonly being denoted as the fracturing pressure of the subject rock(s), and the fracturing pressure commonly being recalculated, and expressed in terms of, an equivalent fracture gradient of the subject rock(s).\",\n \"During drilling, upon approaching one or several formation layers with expected overpressures), the specific gravity/pressure gradient of the drilling fluid is increased to an extent necessary to withstand said overpressure(s).\",\n \"Thus, potentially overpressured formation fluids are prevented from flowing into the well upon drilling into, potentially after having drilled into, said layer(s).\",\n \"If said increase in the pressure gradient of the drilling fluid exceeds the fracture gradient of one or more of the penetrated rocks, the rocks(s) will be fractured and fractures develop in the rock(s).\",\n \"Then, drilling fluid may flow unobstructedly out (leak) from the well and into the fractures, thereby causing the height of the drilling fluid column, and thus the liquid pressure in the liquid column, to be lowered.\",\n \"By so doing, the formation pressure barrier brought about by the drilling fluid pressure exerted in the well is impaired, and this results in the establishment of an undesired, and potentially very dangerous, situation in the well.\",\n \"In order to prevent such fracturing it is often absolutely necessary to isolate the penetrated formation layers from pressures that may fracture the rocks contained therein.\",\n \"As mentioned, such a fracturing pressure may be exerted by the pressure, of the drilling fluid column, but the fracturing pressure may also be exerted by the overpressured formation fluids of other formation layers, usually deeper formation layers, which are being penetrated by the well during drilling.\",\n \"Moreover, and pertaining to an open hole-section, it is the rock(s) of the shallowest part of the section, immediately underlying the casing shoe of the preceding casing, that generally, but not necessarily, is/are the weakest by strength, and thus being the one(s) which may first be fractured.\",\n \"After having started the drilling of a new hole section in a well, it is for this reason common practise to undertake a so-called formation strength test of the shallowest rocks in said hole section.\",\n \"Such a formation strength test is usually carried out immediately after having drilled the uppermost rocks along a 5-10 metre hole length of the new hole section.\",\n \"For example, the formation strength test may consist in supplying said rocks with drilling fluid under a gradually increasing liquid pressure, and increasing the liquid pressure until an incipient fracturing of, and an accompanying leakage of drilling fluid into, the rocks is observed, which determines the fracturing pressure/fracture gradient of the rocks.\",\n \"In the petroleum industry such a formation strength test is usually called a “leak-off test”.\",\n \"In another commonly occurring formation strength test, a so-called formation integrity test, said rocks are also supplied with drilling fluid under a gradually increasing liquid pressure, limiting however the fluid pressure increase to a predefined maximum liquid pressure, and where this liquid pressure is considered to be the maximum required drilling fluid pressure to be applied for the new hole section to be drilled down to the desired drilling depths.\",\n \"This maximum liquid pressure is usually smaller than the fracturing pressure of said rocks, thus not fracturing the rocks during this formation strength test.\",\n \"Therefore, a formation integrity test is usually gentler on said rocks and the subsequent drilling operations than a fracturing test.\",\n \"Such formation strength tests therefore provide a good indication as to the magnitude of liquid pressure, or magnitude of the liquid pressure gradient, whereby the drilling fluid may be arranged during the drilling of a hole section in order to avoid fracturing of the accompanying rocks.\",\n \"Said maximum liquid pressure/liquid pressure gradient also limits the further drilling of a hole section to end at a depth at which the fluid pressure of a formation layer approaches said liquid pressure/liquid pressure gradient.\",\n \"Cementing is also employed as a corrective method to prevent/reduce undesired inflow, and thereby also undesired pressure build-up, of a fluid in one or several regions of a well, including undesired fluid inflow through one or several leaking casings surrounding uncemented annuli of the well, the annulus/annuli possibly extending entirely up to the wellhead of the well.\",\n \"The method consists in injecting cement slurry, possibly with the addition of plasticizing agents, gelling agents, stabilisers or other additives, into a relatively short annular interval covering said inflow region(s), whereupon the cement slurry or agent hardens or sets in such a way that it forms a pressure- and flow-preventive barrier which, in principle, shall prevent/reduce such fluid inflows.\",\n \"Disadvantages of the Prior Art Cementing jobs in a well are often encumbered with problems and disadvantages associated with the physical and chemical properties of the cement.\",\n \"At the start of a cementing job, the cement exists in a liquid state as a slurry.\",\n \"Later, and through a time-adapted curing process, the cement slurry is transformed to firm, or hardened, cement.\",\n \"It is therefore of paramount importance that the cement slurry is placed into the intended cavity/void, usually an annulus, of the well while the cement slurry is sufficiently fluidised to enable it to displace onwards into this cavity/void/annulus.\",\n \"Therefore, the placing of cement slurry into the well must be carried out before a significant thickening or hardening or the cement slurry has taken place.\",\n \"If, during the placing into the well, the cement slurry is thickened or hardens prematurely, or if the cement slurry is introduced into and is thickened/hardened in an incorrect region/interval of the well, the cement will easily cause more problems than what it solves.\",\n \"During the placing into the well, such a premature thickening/hardening of the cement slurry may develop if the slurry unintentionally is supplied with saline water, for example sea water or saline formation water.\",\n \"Upon placing the slurry against a permeable formation layer of a surrounding formation hole wall, a premature thickening/hardening of the cement slurry may also develop in the event that the slurry water phase is being filtered and flows into said permeable formation layer.\",\n \"If the cement slurry is thickened/hardened earlier than planned, thickened/hardened cement unintentionally may be placed in pipes and/or equipment otherwise intended to be open throughout.\",\n \"For example, a premature thickening/hardening of the cement slurry in a cementing pipe and/or in a surrounding casing to be fixedly cemented, may result in the unintentional clogging of said pipes.\",\n \"Correspondingly, a cement slurry being pumped down through, possibly in or around, (a) leaking pipe(s), may result in the unintentional and firm cementing of pipe(s) and/or equipment not to be fixedly cemented, hence resulting in said pipe(s) and/or equipment not working as intended, and possibly in not being able to remove the pipe/equipment from the well if or when this should become necessary.\",\n \"For example, leakages in said cementation pipe and/or in the surrounding casing, may result in the cement slurry unintentionally being conducted onwards to the annulus between the outside of the cementing pipe and the surrounding casing, resulting in the cementing pipe unintentionally being fixed in said annulus upon thickening/hardening of the cement slurry.\",\n \"In the worst case, such unintentional occurrences may result in having to re-drill all or parts of the well.\",\n \"Said pipe leakages may also result in the cement slurry not displacing sufficiently far out/up into the relevant annulus to be fixedly cemented, which subsequently may result in the cement not exhibiting the desired pressure- and flow-preventive effect in the annulus.\",\n \"In connection with such cementing jobs, channel-formed cavities/voids in the cement, so-called channelling in the cement, commonly develop, especially when cementing long pipe sections.\",\n \"Such channelling in the cement represents an undesired effect that may result from the cement slurry and an accompanying liquid front between the cement slurry and an overlying drilling fluid, are being exposed, among other factors, to an uneven laminar flow while the slurry is being displaced out/up into an annulus in the well.\",\n \"Such an uneven laminar flow often results in an insubstantially uniform and inefficient displacement of said liquid front in the annulus, drilling fluid channels thus being formed in the inflowing cement slurry as it flows out/up into the annulus, and resulting in said channels being maintained permanently in the annulus after hardening of the cement slurry.\",\n \"Said annulus may be an annulus between two casings and/or an annulus between one casing and a surrounding formation hole wall.\",\n \"Such channel-shaped cavities/voids in the cement often cause pressure- and fluid leakages.\",\n \"Such pressure- and fluid leakages may also develop in connection with the curing process of the cement slurry.\",\n \"initially, during the curing process, cement nuclei are formed, increasing gradually into a sufficiently large number to form a continuous lattice structure of cement nuclei, the lattice structure being sufficiently strong to carry the weight of newly formed cement nuclei.\",\n \"At this stage of the curing process, when the load supporting lattice structure is established, and before consuming and chemically bonding the water phase of the cement slurry during the curing process, said water phase exists as an independent liquid in said lattice structure, the water phase at this stage of the curing process being exposed only to hydrostatic liquid pressures of its own and overlaying liquids.\",\n \"However, the hydrostatic liquid pressure of the water phase is substantially less than the hydrostatic liquid pressure of the original cement slurry.\",\n \"This reduction in hydrostatic liquid pressure may be sufficiently large for potential overpressured formation fluids of fluid-communicating formation layers to flow into the hardening slurry, causing subsequent pressure- and fluid leakages through it.\",\n \"The presence of such formation fluids in the hardening cement slurry may prevent a further chemical reaction between cement and water in such a way that the function of the cement as a pressure- and flow-preventive barrier in the well, is impaired or destroyed.\",\n \"It is obvious, however, that such impairments of the cement in a well may result in the overlying rocks thus not being sufficiently protected against pressure conditions that may cause fracturing of the rocks.\",\n \"The fluid pressure from overpressured formation layers may thus propagate, via one or several annuli in the well, further upward in the well and, for example, cause an unintentional pressure build-up at the wellhead of the well.\",\n \"At worst, such pressure- and fluid leakages may lead to an uncontrolled outflow of overpressured formation fluids at the surface of the well, a so-called surface blow-out; or to overpressured formation fluids flowing, via the well, between different formation layers, a so-called underground blow-out.\",\n \"Furthermore, hardened cement, in the manner used in a well, constitutes a stiff, brittle and substantially inflexible material possessing a relatively large shear strength.\",\n \"Advantageously, in some areas of utilisation, such material properties may be exploited.\",\n \"For example, cement may be used as a load transferring connection between a surface casing and its surrounding formation hole wall.\",\n \"As mentioned, in a borehole wall consisting of fractured or unconsolidated rocks, cement may be used as a shear-sustaining binding material binding together loose rocks and preventing the rocks from falling out from the hole wall and into the accompanying hole.\",\n \"During the drilling of a hole section, such a falling out of loose rock fragments may cause large technical drilling problems if, for example, such unfastened rock fragments firmly pack around a drill string and prevent or stop any further drilling.\",\n \"In other areas of utilisation, however, such material properties may appear less advantageous.\",\n \"Some reservoirs consist, for example, of very porous sedimentary rocks, for example chalk or unconsolidated sand, such rocks often being soft and exhibiting very little material strength.\",\n \"In deeper layers of the subsurface, such porous rocks are usually overpressured, which overpressure is and, through geologic time, has been a necessary prerequisite in order to preserve the porosity of a rock during its course of compaction.\",\n \"In the process of recovering formation fluids from such a porous and weak reservoir rock, the formation pressure decreases gradually.\",\n \"Consequently, a corresponding and gradual compression (compaction) of the rock pores also will take place, resulting in accompanying vertical movements in the reservoir rock and in the overlaying rocks.\",\n \"However, well pipes, for example casings and/or liners, being placed in and throughout such compacting reservoir rocks are relatively.\",\n \"rigid and not of a physical state such that they, as for the rock pores, may be pressed together, thereby compensating for vertical movements in the reservoir.\",\n \"Consequently, relative movements take place between the well pipes and the surrounding rocks, and where the relative movements will tend to bend out/deflect, buckle/break and/or twist the pipes.\",\n \"Moreover, and due to the stiffness, shear strength and compressive strength of the cement, cementing such well pipes to the surrounding rocks will further tend to prevent this bending/deflection, buckling and/or twisting.\",\n \"Sufficiently large stress concentrations may thus be generated in the well pipes for one or several well pipes, in one or several places, to be torn to pieces or to be severely deformed.\",\n \"Such a destruction or deformation of one or several well pipes may result in a production well becoming completely or partially abandoned, or in having to drill a new production well, thus incurring large technical and economical disadvantages.\",\n \"The method of injecting cement slurry, possibly plasticizers, gelling agents, stabilisers or other additives into a relatively short annulus interval covering one or several undesired inflow area(s) in a well, is also encumbered with channels being formed and subsequent accompanying pressure- and fluid leakages in the cement.\",\n \"Furthermore, production-related and relative pipe movements may also cause the cement to fracture or to unfasten from surrounding well pipes, and causing thus the cement to begin leaking.\",\n \"Usually, such a cementing procedure will therefore only provide a usable pressure- and flow seal for a short period of time, after which problems of pressure build-up and potential fluid leakages may reappear in the well.\",\n \"THE OBJECTIVES OF THE INVENTION One definite objective of this invention is to provide a new method for pressure- and flow-preventive fixing of pipes, for example casings and liners, and possibly accompanying equipment in a well.\",\n \"Another definite objective of the invention is to be able to use the method in a completed well for the purpose of placing one or several pressure- and flow-preventive barriers in one or several annuli of which at least one pipe thereon is leaking.\",\n \"The primary objective, however, is to be able to use the method, completely or partially, to replace the prior art functions of cement in a well, concurrently avoiding or reducing the above-mentioned problems and disadvantages associated with well cementing.\",\n \"Achieving the Objectives Instead of placing cement slurry in the relevant cavity/void of the well, usually an annulus, the objectives are achieved by placing along a sufficient length of said cavity/void/annulus, a fluidised mixture of unconsolidated matter.\",\n \"During the placing, the mixture of unconsolidated matter must be sufficiently fluidised for the mixture to displace onward to, and sufficiently far into, the cavity/void/annulus of interest.\",\n \"In most applications, the placing may be carried out in the simplest and most efficient way by pumping the mixture of unconsolidated matter, as for cement slurry, through a connecting pipe onward to and into said cavity/void/annulus of the well.\",\n \"In this respect, pipes and equipment of the types known in the art for fixedly cementing pipes in a well, may be used.\",\n \"Largely, prior art methods for fixedly cementing such well pipes, may also be used for placing said mixture of unconsolidated matter into said cavity/void/annulus of the well.\",\n \"Furthermore, and for the purpose of providing the fluidised mixture of unconsolidated matter with rheological properties enabling the mixture to be placed in the well, knowledge in the field of rheology together with devices, methods and additives which, for example, are used in the preparation and handling of drilling fluids/well cement, may be employed.\",\n \"In order for such a mixture of unconsolidated matter to work as a pressure- and flow-preventive barrier in the well, the mixture of unconsolidated matter, being a substitute for cement, must be arranged in such a way in the well that it, when the fluidised unconsolidated matter has set in its operational position in the cavity/void/annulus, exhibits sufficiently good pressure- and flow-preventive properties.\",\n \"In the method according to this invention, a mixture of unconsolidated matter comprised by naturally occurring and/or synthetically manufactured granular material, is therefore used in said barrier.\",\n \"In the operation position in said cavity/void/annulus, the granular particles are assembled in such a way that they exhibit a very small permeability towards a fluid flowing through the mixture of unconsolidated matter.\",\n \"Consequently, this method presupposes that the pressure- and flow-preventive barrier of unconsolidated matter is permeable, and that said fluid thereby leaks through the barrier of unconsolidated matter.\",\n \"If the barrier of unconsolidated matter is arranged with a sufficiently small permeability over a sufficiently long length interval in the well, and a fluid is flowing through the barrier of unconsolidated matter, the fluid in the barrier of unconsolidated matter, however, will be exposed to a large flow resistance (flowing pressure drop) and thereby move very slowly (very small flow velocity) through the barrier of unconsolidated matter, and in such a way that the corresponding through-put flow time of the fluid theoretically may extend to several tens of thousands of years or more.\",\n \"This course of flow is influenced by different parameters in accordance with Darcy's law which expresses a relation between several parameters and the flow velocity of a fluid when the fluid flows through a porous and permeable material; in which: v=k(Pin−Pout)/(μ·L); wherein ‘v’—flow velocity of the fluid (cm/sec), ‘k’—permeability of the material (Darcy), ‘Pin’—upstream pressure potential of the fluid (atmospheres), ‘Pout’—downstream pressure potential of the fluid (atmospheres), ‘(Pin−Pout)’—pressure loss through the material (atmospheres), ‘μ’—viscosity of the fluid (centipoise) ‘L’—length of permeable material (cm).\",\n \"Considering that the fluid flow time through the barrier of unconsolidated matter theoretically is in the order of thousands of years, it is evident that the subsequent fluid leakage (amount of fluid leaking through the barrier) will be extremely small and, for practical purposes, negligible.\",\n \"On the other hand, by using a cement barrier in a well, large pressure- and fluid leakages through the cement barrier often appear and are observed.\",\n \"In the abovementioned perspective of time, however, such a cement barrier may constitute a substantially poorer, less enduring and insubstantially ductile/flexible barrier against pressure and flow than that of a barrier of unconsolidated matter.\",\n \"The permeability ‘k’ of the mixture of unconsolidated matter and the extent or length ‘L’ of the barrier of unconsolidated matter in the well represent those parameters of Darcy's law which most readily may be influenced and controlled for the purpose of obtaining a sufficiently small fluid flow velocity ‘v’ through the barrier of unconsolidated matter.\",\n \"Also, and to a lesser degree, the flow velocity ‘v’ may be influenced and controlled by selecting a suitable downstream pressure potential ‘Pout’ for the flowing fluid.\",\n \"In practice, ‘Pout’ is comprised by the hydrostatic pressure being exerted on the barrier of unconsolidated matter by an overlaying/shallower liquid column, for example a water column, which hydrostatic pressure may be adapted, to some extent, by changing the specific gravity of the liquid column.\",\n \"The upstream pressure potential ‘Pin’ of the fluid, however, usually is comprised by the formation pressure being exerted on the barrier of unconsolidated matter from an underlying/deeper reservoir layer, which pressure substantially may not be influenced/controlled, or may not be desired to be influenced/controlled, in consideration of said fluid flow velocity ‘v’ through the barrier of unconsolidated matter.\",\n \"On the other hand, there may exist a desire to influence/control said formation pressure ‘Pin’ in consideration of the exploitation progress and degree of recovery of a reservoir, for example by implementing in the pertinent reservoir(s), actions of artificial stimulation, including water flooding.\",\n \"According to Darcy's law, the permeability ‘k’ of the mixture of unconsolidated matter is proportional to the fluid flow velocity ‘v’ and, consequently, inversely proportional to the fluid through-put flow time, while the length ‘L’ of the barrier of unconsolidated matter is inversely proportional to the flow velocity ‘v’ and, consequently, proportional to the fluid through-put flow time.\",\n \"By so doing, the fluid flow velocity ‘v’ and also through-put flow time may be controlled by selecting a suitable permeability ‘k’ and/or barrier length ‘L’.\",\n \"In practice, considering that the maximum barrier length ‘L’ is limited to the length of the pertinent cavity/void/annulus of a well, the largest scope of influence/control on the flow velocity ‘v’/through-put flow time is gained by arranging the mixture of unconsolidated matter in such a way that it, in the operational position, exhibits a suitable permeability ‘k’.\",\n \"The physical and chemical conditions prevailing in the subsurface layers of the individual well may vary from well to well.\",\n \"Among other things, such physical and chemical conditions include reservoir depth, formation pressure(s) and temperature(s), type(s) of formation fluid(s) inclusive of its/their chemical compositions and physical properties, including properties or conditions influencing the viscosity of the fluid(s).\",\n \"Considering that the prevailing physical and chemical conditions may vary from well to well, the permeability that is considered to be suitable for the pertinent well, also may vary from well to well.\",\n \"Darcy's law shows, among other points, that the fluid viscosity ‘μ’ is inversely proportional to the fluid velocity ‘v’.\",\n \"In one specific barrier of unconsolidated matter being arranged with a one specific permeability, a gas possessing a small viscosity, for example, will flow much faster through the barrier of unconsolidated matter than a heavy crude oil possessing a large viscosity is able to do in the same barrier.\",\n \"In the event of wanting the gas and the crude oil to flow with equal flow velocity through each their own barrier of unconsolidated matter of equal (flow through-put) length, the barrier of unconsolidated matter for the gas must therefore be arranged with a substantially smaller permeability than that of the barrier of unconsolidated matter for the crude oil.\",\n \"A barrier of unconsolidated matter should be arranged such that it, in the operational position, exhibits a permeability in the order of preferably, but not necessarily, a few milliDarcy (mD) and down to a level of microdarcy (μD), for example 0.001 mD (=1 μD).\",\n \"In most wells, these are permeability values that will provide the desired pressure- and flow-preventive effect.\",\n \"Nevertheless, for reasons mentioned above, the specific permeability of the barrier should be evaluated and determined based on the prevailing conditions in the pertinent well.\",\n \"The mixture of unconsolidated matter is arranged with the desired permeability by it being composed of, and in the operational position consisting thereof, mixed granular particles of at least one particle size and, preferably, of several particle sizes.\",\n \"Packed together, the permeability of the mixture of unconsolidated matter is determined by the geometric shape of a pore network comprised by the pores of the mixture of unconsolidated matter and their mutual pore connections.\",\n \"The degree of variation in particle sizes has a large impact on the tightness at which the unconsolidated matter particles may be packed, which largely influences how the pore network will appear, hence also what the permeability of the mixture of unconsolidated matter will be.\",\n \"Also, the general particle size of the mixture of unconsolidated matter are of great importance in determining how large the said pores and their mutual pore connections will be, which directly influences the permeability of the mixture of unconsolidated matter.\",\n \"Consequently, one of two methods may be used to affect the permeability of such a mixture of unconsolidated matter; either by the mixture of unconsolidated matter being comprised of different particle sizes, or by the mixture of unconsolidated matter being comprised of small particles of a relatively homogenous size.\",\n \"The distribution of particle sizes in such a mixture of unconsolidated matter is often expressed by the concept of sorting.\",\n \"The concept of sorting is a qualitative measure of the degree of variation, or the range of variation, of different particle sizes in the mixture of unconsolidated matter.\",\n \"A poorly sorted mixture of unconsolidated matter may include a large spectrum of particle sizes, for example particles in the size ranges of gravel, sand, silt and clay.\",\n \"In comparison, a moderately sorted mixture of unconsolidated matter may include a small spectrum of particle sizes, for example medium sand and fine sand, while a very well sorted mass may include only one relatively homogenous particle size, for example coarse silt.\",\n \"In the packed condition, such a poorly sorted mixture of unconsolidated matter may exhibit a very small permeability.\",\n \"A very well sorted mixture of unconsolidated matter consisting of coarse silt may exhibit an equivalent small permeability, while a very well sorted mixture of unconsolidated matter consisting of very coarse sand may exhibit a very large permeability.\",\n \"Such a specification of sorting, however, is imperfect for quantifying, or for specifying the quantities of, the different particle sizes comprising the mixture of unconsolidated matter.\",\n \"On the other hand, the distribution of particle sizes in the mixture of unconsolidated matter may better be described and quantified by means of, for example, statistical concepts, wherein the distribution of particle sizes in the mixture of unconsolidated matter may be described by means of a cumulative distribution function.\",\n \"In practice, different particle sizes may be provided by, for example, sieving and grouping naturally occurring granular unconsolidated matter into several different particle size categories.\",\n \"Each such category is comprised of particles of a particular particle size range, the particle size range of each category differing from particle size ranges of possibly other categories.\",\n \"Alternatively, synthetically manufactured granular material made of particle sizes within the pertinent particle size categories may be used.\",\n \"Subsequently, certain amounts of particles of each of the pertinent particle size categories are assembled and mixed together, the mixture of unconsolidated matter thus being arranged with one particular distribution of particle sizes, hence one particular pore network shape of mixture of unconsolidated matter, which provides one particular permeability for the mixture of unconsolidated matter when placed in the operational position as a pressure- and flow-preventive barrier in the well.\",\n \"Several scales exist specifying the different particle size categories, and the preferred scale may be related, largely, to particular trade disciplines.\",\n \"The so-called Udden-Wentworth particle size scale and the so-called Krumbein phi (φ) particle size scale are generally known and are used, for one thing, in geologic disciplines, for example in sedimentology.\",\n \"In the construction industry and in geotechnical environments, among other-matters, it is common, however, to use a scale referring to the mesh size (grate size) of a sieve device, for example the commonly used and so-called American Society of Testing and Materials (A.S.T.M.)\",\n \"Sieve Scale.\",\n \"The scale specifies particle size categories referring to so-called “mesh” sizes.\",\n \"For example, a 200 mesh size represents 0.074 mm large sieve openings in an accompanying screen-cloth or grate of the sieve device.\",\n \"Similar scales and/or concepts also exist which, to varying degrees, are used in different geographical regions and/or trade disciplines.\",\n \"In the Udden-Wentworth scale, particles are grouped in particle size categories on the basis of average particle diameter specified in millimetres.\",\n \"Examples of such size categories are fine gravel/granules (2-4 mm), very coarse sand (1-2 mm), coarse sand (0.5-1 mm), medium sand (0.25-0.5 mm), fine sand (0.125-0.25 mm), very fine sand (0.0625-0.125 mm), four categories of silt (0.0039-0.0625 mm), and also clay particles (<0.0039 mm).\",\n \"In the Krumbein phi (φ) scale, the particle sizes are converted to φ-values, in which: φ=log2 d; wherein ‘d’—average particle diameter (mm).\",\n \"Expressed in Krumbein φ-values, the preceding Udden-Wentworth examples of particle size categories may be specified as fine gravel/granules (φ=−2 to −1), very coarse sand (φ=−1 to 0), coarse sand (φ=0 to +1), medium sand (φ=+1 to +2), fine sand (φ=+2 to +3), very fine sand (φ=+3 to +4), four categories of silt (φ=+4 to +8), and also clay particles (φ=+8 or more).\",\n \"Each particle size category being specified as integer φ-values, and not in fractions or decimal numerals, as in the Udden-Wentworth scale, such Krumbein φ-values are more easily treated statistically.\",\n \"When using the Krumbein phi (φ) scale, the distribution of particle sizes in a mixture of unconsolidated matter (the sorting of the mixture of unconsolidated matter) is commonly specified as the range of variation (in φ-values) which includes an amount of particles comprising approximately ⅔ of all particles in the mixture of unconsolidated matter.\",\n \"Statistically, this range of variation constitutes two times the standard deviation of the unconsolidated matter particles, and the standard deviation is therefore a commonly accepted measure of the sorting of a sediment or a mixture of unconsolidated matter.\",\n \"In the construction industry and in geo-technical environments, among other things, it is customary to quantify a particular distribution of particle sizes (sorting) of a mixture of unconsolidated matter by means of a so-called sieve curve.\",\n \"Specified in sieve- or mesh sizes, the sieve curve specifies the relative amounts, or the mass ratio, of the pertinent particle size categories which constitute, or is to constitute, the mixture of unconsolidated matter.\",\n \"For example, the patent publication U.S. Pat.\",\n \"No.\",\n \"5,417,285 describes the use of a short plug of particulate material in connection with a partition or obstruction member in a well, the partition/obstruction member typically consisting of a mechanical plug, for example an inflatable packer or a so-called bridge plug.\",\n \"Concurrently, and concerning the composition of the short particulate material plug, the publication describes seven different mixtures of particulate material and their specific and different particle compositions, each particle composition being expressed by means of “mesh” particle size categories and fractions by weight in percentages of the total weight of each mixture.\",\n \"At the same time, the permeability of each mixture of particulate material has been determined by testing and specified in the publication.\",\n \"Three of the said mixtures of particulate material showed especially a small permeability, and their particle compositions and permeabilities are of such a nature that they are considered suitable in a barrier of unconsolidated matter of the type comprised by this invention.\",\n \"The particle composition and permeability of the three mixtures are stated in the following table summary: Test mixture - ref.\",\n \"test-number cited in the publication Particle size 7 8 9 category Fraction by weight of the (mesh sizes) total weight of the mixture(%) 20/40 mesh sand 60 60 60 100 mesh sand 20 15 20 200 mesh sand 15 15 15 Bentonite “gel” 5 10 5 (clay fraction) Permeability (mD) 0.064 0.063 0.081 Said patent publication describes a 20/40 mesh sand as a conventional coarse sand, a 100 mesh sand as a conventional medium (“intermediate”) sand and a 200 mesh sand as a conventional fine sand, the test mixtures 7-9 also containing a particle fraction of fines described as a gel of bentonite/clay particles.\",\n \"Chemically, the sand particles are described as consisting of preferably silica (silicon dioxide), mineralogically denoted as quartz.\",\n \"Also, this is a suitable choice in that quartz (silicon dioxide) is one of the most weathering resistant minerals to be found in nature, and quartz/silica (silicon dioxide) therefore ought to provide excellent time- and weathering resistance in a well.\",\n \"Moreover, the proprietors of the present invention have carried out laboratory experiments involving a similar mixture of unconsolidated matter.\",\n \"Within a period of time of ca.\",\n \"1.5 months, and by means of measurements, the permeability of the mixture of unconsolidated matter, among other things, was calculated as a function of the settling, or compaction, of the unconsolidated matter particles.\",\n \"Also, the experiments confirmed that it is possible, in practice, to produce a fluidised mixture of unconsolidated matter which is easy to pump.\",\n \"In the mixture of unconsolidated matter (predominantly silica/quartz), ca.\",\n \"80 percent by weight of the mass consisted of sand size particles in the particle is size categories coarse sand (0.5-1 mm), medium sand (0.25-0.5 mm), fine sand (0.125-0.25 mm) and very fine-sand (0.0625-0.125 mm), while ca.\",\n \"20 percent by weight of the mass consisted of silt size particles in the particle size range of 0.0039-0.0625 mm, half of which (ca.\",\n \"10 percent by weight) in the size range of 0.005 mm (fine silt).\",\n \"The last-mentioned fine silt particles were added to the mixture of unconsolidated matter exclusively to act as a permeability-reducing filler of the pores in the mixture since this fraction of mixture fines only contains negligible amounts of clay particles.\",\n \"This differentiates this mixture of unconsolidated matter from the three mixtures specified in the preceding table, wherein relatively large fractions by weight of clay particles, so-called bentonite gel, are used in the mixtures, such suspended clay particles in the pores of the mixture acting as a binder between the particles.\",\n \"Initially, a 1 metre length of mixture of unconsolidated matter was placed in the bottom of a vertically positioned plastic pipe, 6 metres long in total, after which the entire pipe was filled with fresh water.\",\n \"During the subsequent time period of ca.\",\n \"1.5 months, regular measurements were taken whereupon the permeability of the mixture of unconsolidated matter was calculated for the time period, observing during the time period diminishing permeability values.\",\n \"At expiry of the time period, and after settling in the plastic pipe, the mixture could exhibit a permeability of 0.001 mD (=1 μD).\",\n \"Furthermore, when being placed in the pipe, the mixture of unconsolidated matter was fluidised and contained ca.\",\n \"83 percent by weight of unconsolidated matter particles and ca.\",\n \"17 percent by weight of liquid, which consisted of ca.\",\n \"11 percent by weight of water and ca.\",\n \"6 percent by weight of a suitable plasticizer.\",\n \"The plasticizer was used in order to avoid uneven settling of the fine-grained and coarse-grained particle fractions of the fluidised mixture of unconsolidated matter, but also in order to maintain the largest possible fraction of unconsolidated matter, hence the smallest possible fraction of liquid, in the fluidised mixture of unconsolidated matter.\",\n \"Lignosulphonate represents one example of a common plasticizer/viscosity-regulating agent being used, for instance, in the petroleum industry, for example when preparing drilling fluids.\",\n \"Moreover, these are merely examples of how such a mixture of unconsolidated matter may be composed, and of how the mixture of unconsolidated matter may be fluidised.\",\n \"Further specifications of compositions of mixtures of unconsolidated matter, and also specifications of specific substances and agents, devices and methods known in the art for fluidising the mixture, are considered to be of prior art technical nature provided the method according to this invention is brought forward to a person skilled in art.\",\n \"After having assembled and mixed in specific quantities of the pertinent particle size categories, for example as specified in the table mentioned above, and in such a way that the mixture of unconsolidated matter thus has been arranged with a distribution of particle sizes which, in the operational position, is to provide the desired permeability, the mixture of unconsolidated matter is fluidised prior to placing it in the relevant cavity/void/annulus of the well, this fluidisation simplifying the placing of the mixture of unconsolidated matter in the well.\",\n \"For example, the fluidisation may be carried out by means of prior art devices and methods for stirring and mixing fluids and/or solids.\",\n \"In this fluidisation process, the mixture of unconsolidated matter is mixed together with a suitable carrier fluid to become a fluidised mixture of unconsolidated matter, the fluidised mixture of unconsolidated matter being arranged in such a way that it thereafter, and preferably, may be pumped, for example by means of powerful cementing pumps and -equipment of the kind typically being used during the cementing of well pipes.\",\n \"Most simplistically, the carrier fluid may be comprised of water.\",\n \"On the other hand, plasticizers, gelling agents, stabilisers, weighting materials or other additives may be added in order to arrange the fluidised mixture of unconsolidated matter with appropriate physical and/or chemical properties, including rheological properties, to enable the mixture of unconsolidated matter to be placed and used as intended in the well.\",\n \"For example, the fluidised mixture of unconsolidated matter must be arranged with a viscosity enabling the pumping of the mixture through, for example, said cementing pumps/-equipment and connected pipes in the well so that the fluidised mixture of unconsolidated matter may be displaced further out/up into the pertinent cavity/void/annulus of the well.\",\n \"Also, and by example, the mixture may be arranged with suitable thixotropic properties.\",\n \"Furthermore, the carrier fluid should constitute a minimal weight fraction, suitable for the purpose, of the mixture of unconsolidated matter.\",\n \"Consequently, the fluidised mixture is comprised of a maximum weight fraction of unconsolidated matter particles forming the barrier of unconsolidated matter in the well, and this measure prevents or restricts a potential formation of surplus liquid originating from the carrier fluid.\",\n \"During pumping, possibly after the pumping and in connection with the settling of the mixture of unconsolidated matter in said cavity/void/annulus, such measures advantageously result in avoiding or reducing premature settling (segregation) of possible coarse-grained fractions and, hence, in segregating these from remaining suspended finer-grained particle fractions in the fluidised mixture of unconsolidated matter.\",\n \"A mixture of unconsolidated matter may thus be placed in the operational position in the well, a mixture which, after said settling, still is provided with the desired distribution and packing of particle sizes, thus also being provided with the desired permeability.\",\n \"Such a potential uneven settling of particle sizes in the mixture of unconsolidated matter, as a barrier of unconsolidated matter in the well, may result in it exhibiting an uneven permeability distribution along its longitudinal extension in the well, and in the barrier of unconsolidated matter not exhibiting the desired pressure- and flow-preventive effect in the well.\",\n \"Moreover, and in connection with the placing of the unconsolidated matter, care must be exercised in ensuring that the fluidised mixture of unconsolidated matter is arranged with a specific gravity not exceeding the fracturing pressure/fracture gradient of the relevant hole section of the well.\",\n \"In this context, the fluidised and unconsolidated mixture possibly may be arranged with a specific gravity in the order of 2.1, which specific gravity does not differ from typical specific gravity values of a cement slurry.\",\n \"Contrary to cement, such a barrier of unconsolidated matter may not harden in the cavity/void/annulus of the well.\",\n \"In the operational position, the barrier of unconsolidated matter therefore may exhibit plastic properties, inasmuch as the barrier of unconsolidated matter is flexible and ductile and, simultaneously, possibly may exhibit insignificant tensile- and shear strength.\",\n \"After having pressure- and flow-preventively fixed by means of the barrier of unconsolidated matter a specific casing size, these properties of such a barrier of unconsolidated matter must be considered in the event of commencing thereafter the drilling of a subsequent and deeper hole section.\",\n \"If said casing size, entirely from the casing shoe and up throughout the well, is fixed by means of such a ductile mixture of unconsolidated matter, and due to its insignificant tensile- and shear strength, the unconsolidated matter may easily fall down and into the subsequent hole section when being drilled.\",\n \"For instance, this problem may easily be avoided by placing in a length interval immediately underlying said barrier of unconsolidated matter, and in the same cavity/void/annulus, cement and/or another material possessing tensile- and shear strength, for example a mechanical annulus packer, and which prevents unconsolidated matter from falling down and into said hole section.\",\n \"In practice, and in the very same well pipes, this may be carried out by pumping down into the well, and also out/up into said cavity/void/annulus, a volume of cement slurry concurrent with, and immediately following, the fluidised mixture of unconsolidated matter.\",\n \"Afterwards, when the cement slurry is placed in its operational position and is hardening to form a cement barrier in the well, the mixture of unconsolidated matter will be prevented from falling down and into the subsequent hole section when being drilled.\",\n \"This is shown in the following embodiments of the invention.\",\n \"However, such a cement barrier need not be placed in the deepest casing/liner of the well, inasmuch as no subsequent hole section will be drilled into which the mixture of unconsolidated matter may fall down.\",\n \"Advantags Achieved by the Invention Using one or several such barriers of unconsolidated matter in a well offers considerable advantages relative to the prior art, and relative especially to cementing of well pipes.\",\n \"The placing of such a barrier of unconsolidated matter in a well does not involve, for example, a curing process that may create the above-described accompanying problems and disadvantages of cement, in which premature hardening/thickening of a cement slurry unintentionally may plug well pipes and -equipment, or possibly in unintentionally misplacing cement in the well.\",\n \"Hence, the potential problems that may develop in a cement slurry while, during the curing process, forming a continuous lattice structure of cement nuclei therein, are avoided, the formation of this lattice structure possibly resulting eventually in potential overpressured formation fluids originating from fluid-communicating formation layers flowing into the hardening slurry and inflicting subsequent pressure- and fluid leakages through the resulting cement barrier, hence weakening or destroying a further chemical reaction between water and cement, which results in the pressure- and flow preventive function of the cement in the well being weakened or destroyed.\",\n \"Such effects may not develop in a non-hardening barrier of unconsolidated matter because such a fluidised mixture of unconsolidated matter, during settling, will maintain its original liquid pressure/pressure gradient.\",\n \"The absence of such a curing process, as well as said plastic properties of such a barrier of unconsolidated matter, also may result in the channelling in the fluidised mixture of unconsolidated matter not occurring, or occurring unsubstantially, when placed in the well.\",\n \"Due to the ductility of the mixture of unconsolidated matter, potential channels being formed in the mixture of unconsolidated matter may be pressed together and disappear, completely or partially, during the ensuing settling period of the unconsolidated matter particles.\",\n \"Potential fluids, for example drilling fluid, that have been trapped in such channels may thereby be displaced, completely or partially, out of the mixture of unconsolidated matter and not, as for a hardening cement slurry, inflict functional disturbance on the barrier in the well.\",\n \"Also in posterity, awhile being placed as barrier of unconsolidated matter in a well, such a mixture of unconsolidated matter may retain its plastic properties and ductility.\",\n \"Any movements and displacements that, during time, may take place in the surrounding rocks of the well, for example earth quake movements or movements in compacting reservoir rocks, may thus, as for a cement barrier, inflict onto a barrier of unconsolidated matter and accompanying well pipes stresses and accompanying relative movements.\",\n \"As opposed to a cement barrier or a mechanical plug, the ductile barrier of unconsolidated matter, however, may be shaped according to, and adapt to, said relative movements without fractures and subsequent pressure- and fluid leakages being formed therein, and without substantially changing the small permeability of the barrier of unconsolidated matter.\",\n \"Such movements potentially may cause the permeability of the barrier of unconsolidated matter to be further lowered, inasmuch as such influencing forces, in addition to the Earth's force of gravity, may contribute to packing more closely the particles of unconsolidated matter, and thus causing said permeability to be lowered.\",\n \"Hence, the ductility and relative mobility of the barrier of unconsolidated matter cause well pipes to move in such a ductile mixture of unconsolidated matter, the well pipes being fixed by means of such a barrier of unconsolidated matter in the well wherein the well pipes are exposed to said stresses and relative movements.\",\n \"By so doing, such well pipes may be exposed to a substantially larger relative movement in the form of bending, buckling and/or torsion than that or a cement barrier or a mechanical plug before one or several well pipes are torn to pieces or become severely deformed.\",\n \"Advantageously, such a fluidised mixture of unconsolidated matter also may be injected into uncemented cavities/voids/annuli of a well being exposed to undesired inflow of fluids through one or several surrounding and leaking casings/liners.\",\n \"The injection may be carried out, for example, through suitable perforations in a lower part of the casing/liner of the well, or via coiled tubing placed in an upper part of the pertinent annulus of the well.\",\n \"The fluidised mixture of unconsolidated matter is placed in a suitable position in, and in a sufficient well length of, the pertinent cavity/annulus, for example in the entire length of the cavity/void/annulus, thus preventing/reducing pressure- and fluid leakages through said cavity/void/annulus, and without using, for example, cement and/or mechanical packers in the cavity/void/annulus.\",\n \"By so doing, the lifetime of a leaking production tubing in a well may be extended in stead of having to re-complete or abandon the well.\",\n \"Also, well pipes fixed by means of one or several such barriers of unconsolidated matter in a well provide for a significantly easier sidetracking or plugging, permanently or temporarily, of the well.\",\n \"This is due to the unconsolidated matter of the barrier readily being removed in posterity, for example by flushing or circulating out the unconsolidated matter by means of a suitable liquid.\",\n \"This substantially differs from the time-, equipment- and work consuming efforts being initiated in connection with the removal or drill-through of cement emplaced in connection with casing(s)/liner(s) of a well.\",\n \"This is illustrated further in the following embodiments of the invention.\",\n \"Relative to the prior art, including well cementing, the above-mentioned advantages show that the method according to the present invention provides a substantially cheaper technical solution which, moreover, is considerably simpler, more flexible and durable with time with respect to preventing/reducing pressure- and fluid leakages in a well, primarily in connection with the fixing of casing(s)/liner(s) of the well.\",\n \"Also, the method may be used both in vertical, deviated and in horizontal wells.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS In the following part of the description, referring to FIGS.\",\n \"1-5, three non-limiting embodiments of the method according to the invention will be shown, one specific reference numeral referring to the same detail in all figures is where this detail is indicated, wherein: FIG.\",\n \"1 and FIG.\",\n \"2 show schematic vertical sections through a hole section of a well, in which hole section a casing is placed, and FIG.\",\n \"1 shows a fluidised mixture of unconsolidated matter placed in said casing pending displacing of the mixture of unconsolidated matter out and up into an annulus surrounding the casing, while FIG.\",\n \"2 shows the casing fixed in the hole section by means of the mixture of unconsolidated matter after it having been displaced out and up into said annulus, the mixture being placed as a pressure- and flow-preventive barrier of unconsolidated matter in the annulus; FIG.\",\n \"3 and FIG.\",\n \"4 also show schematic vertical sections through a segment of the hole section shown in FIG.\",\n \"2, the casing of the hole section being fixed in the well by means of the said barrier of unconsolidated matter in the surrounding annulus of the casing, and the figures show measures necessary in order to make a sidetrack of the well departing from said hole section, FIG.\",\n \"3 showing perforation of the casing prior to a subsequent injection of cement slurry, while FIG.\",\n \"4 shows a cut-through casing through which an introductory and new sidetrack hole section of the well is shown also; and FIG.\",\n \"5 shows a schematic vertical section through several consecutive hole sections of a well, each hole section being provided with each their own casing size, and all casing sizes being fixed in the well by means of a barrier of unconsolidated matter placed in the surrounding annulus of each casing size.\",\n \"DESCRIPTION OF EMBODIMENTS OF THE INVENTION Knowledges, devices, appliances, equipment, agents, substances and/or methods known in the art not relating to the actual invention, but which nevertheless is/are, or may be, necessary prerequisites in order to practise the invention, will not be described in any detail in the following three embodiments.\",\n \"Among other things, this includes pumping devices/-equipment and accompanying pipes being placed suitably in the well when practicing the invention.\",\n \"Moreover, the figures only show details which are necessary in order to understand and practise the invention, and therefore the figures do not show, for example, a drilling arrangement and accompanying drilling equipment/well equipment etc.\",\n \"The first embodiment is represented by FIG.\",\n \"1 and FIG.\",\n \"2, FIG.\",\n \"1 showing a lower part of a hole section 10 in a subsurface well, the hole section 10 penetrating an underground formation 12.A casing 14 is placed in the hole section 10, and between the casing 14 and the hole section 10 an annulus 16 exists being filled with drilling fluid 18, the drilling fluid 18 also filling a volume at the bottom of the casing 14.Immediately overlying this volume are placed, in consecutive order, a first leading plug 20, a predefined volume of a fluidised mixture of unconsolidated matter 22 according to the previous description, a second leading plug 24, a predefined volume of cement slurry 26 and a trailing plug 28, the overlaying volume of the casing 14 being filled with drilling fluid 18.All plugs 20, 24 and 28 are placed in a pressure-sealing manner against the casing 14.Furthermore, the leading plugs 20 and 24 are arranged with each their own diaphragm 30 and 32 which, in connection with the subsequent pumping and an accompanying displacement of the mixture of unconsolidated matter 22, cement slurry 26 and the plugs 20, 24 and 28 down through the casing 14 and out/up into the annulus 16, are arranged to rupture when the diaphragms 30 and 32 are exposed to a sufficiently large pump pressure.\",\n \"Furthermore, and associated with the diaphragms 30 and 32, each leading plug 20 and 24 is arranged with each their own through-going hole 34 and 36, through which the mixture of unconsolidated matter 22 and the cement slurry 26 may flow when the diaphragms 30 and 32 rupture due to said pump pressure.\",\n \"These conditions have been further described in the preceding description.\",\n \"However, the trailing plug 28 is massive and seats itself onto the second leading plug 24 at the end of the displacement, the second leading plug 24 being placed onto the first leading plug 20, and both leading plugs 20 and 24 being placed with ruptured diaphragms 30 and 32 in these positions.\",\n \"FIG.\",\n \"2 shows the plugs 20, 24 and 28 placed in these positions, and this figure also shows said mixture of unconsolidated matter 22 placed as a pressure- and flow-preventive barrier of unconsolidated matter 38 in the annulus 16, the unconsolidated matter being arranged with a suitable permeability by means of the distribution of particles described above, is arranged with a suitably small permeability, the annulus in a bottom interval of the well also being filled with an underlying hardened cement in the form of a cement barrier 40.The second embodiment is represented by FIG.\",\n \"3 and FIG.\",\n \"4.The embodiment describes measures necessary for sidetracking in the underground formation 12, and by means of prior art drilling equipment, a hole section 42 extending out from the well hole section 10 in the event of the casing 14 thereof being fixed in the annulus 16 by means of pressure- and flow-preventive particles of unconsolidated matter, cf.\",\n \"the barrier of unconsolidated matter 38, FIG.\",\n \"4 showing a segment of the sidetracked hole section 42.Moreover, this figure shows a hole 44 which, in order to enable sidetracking of the well, has been drilled through the casing 14 and the barrier of unconsolidated matter 38.The barrier of unconsolidated matter 38 surrounding the casing 14 exhibits insignificant tensile- and shear strength, and the drilling of a hole 44 through this barrier of unconsolidated matter 38 possibly may cause particles to unfasten and fall into the casing 14.By so doing, a section of length of the barrier or unconsolidated matter 38 being placed overlying the hole 44, may become destroyed, completely or partially, thus discontinuing, completely or partially, the function of the barrier of unconsolidated matter 38 as a pressure- and flow-preventive barrier.\",\n \"This is, however, a problem which may be solved through simple means.\",\n \"Prior to sidetracking, the casing 14 may be arranged with through-going perforations 46, cf.\",\n \"FIG.\",\n \"3, in an area of the hole section 10 overlying the region in which the drilling of the hole 44 for the ensuing sidetrack is desired.\",\n \"Afterwards, a predefined volume of cement slurry is injected through the perforations 46 and into the barrier of unconsolidated matter 38.During the following hardening of the cement slurry, a cement plug 48 possessing substantial tensile- and shear strength is formed in the annulus 16.Thereafter, in a suitable position underlying the cement plug 48, the hole 44 through the casing 14 and the barrier of unconsolidated matter 38 may be drilled, inasmuch as the cement plug 48 prevents particles of unconsolidated matter from unfastening from an overlying part of the barrier of unconsolidated matter 38 and thereinafter falling down and into the casing 14.Then, the sidetracking of the hole section 42 may be carried out, the casing of the section 42 (casing not shown in the figures) possibly also being fixed by means of an equivalent barrier of unconsolidated matter.\",\n \"If the last-mentioned casing is fixed by means of an equivalent barrier of unconsolidated matter, in which this barrier of unconsolidated matter also is to be placed in an associated annulus section overlying the hole 44, it is important to ensure that the corresponding mixture of unconsolidated matter, during the placing thereof in the annulus of the section 42, does not flow out through the hole 44 and down into the underlying pipe volume of the casing 14.Possibly, this problem may be solved by said pipe volume, prior to sidetracking, either being filled by a corresponding mixture of unconsolidated matter (not shown in the figures), or by placing, in a position immediately underlying the hole 44, a mechanical packer plug (not shown in the figure), possibly also by filling a corresponding mixture of unconsolidated matter between the mechanical packer plug and the hole 44, and in the casing 14.The third and last embodiment, represented by FIG.\",\n \"5, shows a sidetracked well in the underground formation 12, the consecutive casing sizes of the well being pressure- and flow-preventively fixed by means of barriers of unconsolidated matter placed in respective and surrounding annuli thereof.\",\n \"In this embodiment, the surface casing 50 of the well is fixed in the underground formation 12 by means of cement 52.Meanwhile, the subsequent first intermediate casing 54 of the well is fixed by means of a barrier of unconsolidated matter 56 and a short underlying cement barrier 58, and the subsequent second intermediate casing 60 is fixed by means of a barrier of unconsolidated matter 62 and a short underlying cement barrier 64, the barriers of unconsolidated matter 56 and 62 extending upward to, or near, the wellhead of the well (not shown in the figure).\",\n \"The production casing 66 of the well is fixed by means of a barrier of unconsolidated matter 68 and a short underlying cement barrier 70, the barrier of unconsolidated matter 68 of this embodiment not extending upward to, or near, the wellhead of the well, but overlapping only a lower length interval of the preceding second intermediate casing 60.If the well is not drilled any deeper than that shown in FIG.\",\n \"5, placing a cement barrier where the cement barrier 70 is shown in the figure is not required, strictly speaking, inasmuch as such a cement barrier 70 is placed in the well merely to prevent particles of unconsolidated matter from falling down and into a subsequent hole section (not shown in the figure).\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473907"}}},{"rowIdx":763,"cells":{"text":{"kind":"list like","value":[["Tire components having improved modulus","A tire component including a tire tread that comprises a vulcanized rubber, a rubber filler, and a modified rubber, where the modified rubber i) contains pendent or functional groups that contain carboxylic acid or anhydride groups, ii) has pendently grafted thereto a polymerized metal salt of an unsaturated carboxylic acid, or iii) contains both i and ii.","The use of such modified rubbers in tire components advantageously increases the G′ of such components."],["1.A tire component including a tire tread that comprises: a vulcanized rubber, an inorganic filler; and a modified rubber that contains i) pendent or terminal functional groups containing carboxylic acid or anhydride groups or ii) a polymerized metal salt of an unsaturated carboxylic acid.","2.A tire component including a tire tread that comprises: a vulcanized rubber, an inorganic filler; a modified polyolefin that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups; and a modified rubber that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups.","3.The tire component of claim 2, wherein the tire component is devoid of zinc oxide.","4.A tire component including a tire tread that comprises: a vulcanized rubber; an inorganic filler; a modified rubber that contains a polymerized metal salt of an unsaturated carboxylic acid, and wherein the tire component is devoid of zinc oxide 5.A tire tread comprising: a vulcanized rubber; from about 10 to about 100 parts by weight inorganic filler per 100 parts by weight rubber; and from about 0.2 to about 10 parts by weight of modified rubber per 100 parts by weight rubber; and wherein said modified rubber contains i) pendent or terminal functional groups containing carboxylic acid or anhydride groups or ii) a polymerized metal salt of an unsaturated carboxylic acid.","6.A tire prepared by a process comprising the steps of: vulcanizing a rubber composition that has been prepared by compounding ingredients comprising: a rubber, an inorganic filler; a modified rubber that contains i) pendent or terminal functional groups containing carboxylic acid or anhydride groups or ii) a polymerized metal salt of an unsaturated carboxylic acid; and optionally a modified polyolefin that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups.","7.The tire component of tread of claims 1-5 or the tire of claim 6, wherein the modified rubber is vulcanized.","8.The tire component or tread of claims 1-5 or the tire of claim 6, where the rubber composition further comprises carbon black.","9.The tire component or tread of claims 1-5 or tire of claim 6, where the inorganic filler comprises silica, aluminum hydroxide, or both.","10.The tire component of claims 1 or 4, where the polymerized metal salt of an unsaturated carboxylic acid derives from an α,β-ethylenically unsaturated carboxylic acid salt having from 3 to 8 carbon atoms.","11.The tire component of claim 10, where the unsaturated carboxylic acid salt contains sodium, potassium, magnesium, calcium, zinc, barium, aluminum, tin, zirconium, lithium, or carbonium.","12.The tire component or tread of claims 1-3 an 5, or the tire of claim 6, where the modified rubber includes functional groups deriving from citraconic acid, cinnamic acid, methacrylic acid, itaconic acid, or mixtures thereof, and maleic anhydride, citraconic anhydride, itaconic anhydride, or mixtures thereof 13.The tire component or tread or claims 1-5 or the tire of claim 6, where the rubber composition further comprises a coupling agent, a dispersing agent, or both.","14.The tire component or treads of claims 1, 4-5, or the tire of claim 6, where the modified rubber is a macromolecule comprising an unsaturated hydrocarbon rubber backbone having pendently grafted thereto a polymerized metal salt of an unsaturated carboxylic acid.","15.The tire component or tread or tire of claim 14, where the unsaturated rubber backbone is a homopolymer of a conjugated diene, a copolymer of a conjugated diene and a vinyl aromatic monomer, or a terpolymer of ethylene, propylene, and a diene.","16.The tire component or tread or tire of claim 12, where the modified rubber contains from about 0.02 to about 28 milliequivalents of functional groups per gram of modified polymer.","17.The tire of claim 6, where the rubber composition comprises from about 1 to about 100 parts by weight inorganic filler phr, from about 0.1 to about 25 parts by weight modified rubber phr, and from about 0.1 to about 25 parts by weight of the modified polyolefin phr.","18.The tire component or tread of claims 1-5, or the tire of claim 6, where the tire is devoid of particulate nylon.","19.The tire component or tread or claims 1-4 or the tire of claim 6, where, the poly(metal carboxylate) graft rubber is prepared by i) dissolving an unsaturated hydrocarbon rubber in a solvent, ii) adding to the dissolved rubber a metal salt of an unsaturated carboxylic acid to form a rubber/carboxylic acid blend, iii) adding to the rubber/carboxylic acid blend a free radical initiator, and iv) heating the reaction mixture to a temperature of from about 40 to about 150° C. for about 0.1 to about 100 hours.","20.A rubber composition comprising: a vulcanizable rubber, a filler; and a modified rubber that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups.","21.The rubber composition of claim 20, where the rubber composition further comprises a modified polyolefin that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups.","22.The rubber composition of claims 20 or 21, wherein the filler is carbon black.","23.The rubber composition of claims 20 or 21, wherein the filler is a mixture of carbon black and silica."],["<SOH> BACKGROUND OF THE INVENTION <EOH>Hysteresis loss and storage modulus are properties of rubber that have a significant impact on tire performance.","Hysteresis loss can be defined by the formula tan δ=G″/G′, where G″ is a measure of the dynamic loss modulus and G′ is the measure of storage modulus.","Generally, as the temperature of a tire increase, G′ decreases.","As a result, there may be a decrease in handling.","Inorganic fillers, such as silica, impart improved wet traction (as demonstrated by a high tan δ at 0° C.) and improved snow traction (as demonstrated by a low tan δ at −20° C.) when used as filler within tire treads.","Mixing silica into a tire stock, however, is difficult because silica particles agglomerate and therefore they are not easily dispersed.","In response, processing and dispersing aids are used during compounding.","Unfortunately, the use of these processing and dispersing aids enhances the decrease in modulus that is typically observed at high temperatures.","Because inorganic fillers are technologically useful, there is a need to overcome the problems associated with the loss of G′ at elevated temperatures in tire treads containing inorganic fillers."],["<SOH> SUMMARY OF THE INVENTION <EOH>In general the present invention provides a tire component including a tire tread that comprises a vulcanized rubber, an inorganic filler, and a modified rubber, where the modified rubber i) contains pendent or functional groups that contain carboxylic acid or anhydride groups, ii) has pendently grafted thereto a polymerized metal salt of an unsaturated carboxylic acid, or iii) contains both i and ii.","The present invention also includes a tire tread comprising a vulcanized rubber, from about 10 to about 100 parts by weight inorganic filler per 100 parts by weight rubber, and from about 0.2 to about 10 parts by weight modified rubber per 100 parts by weight rubber.","The present invention also provides a tire component including a tire tread that comprises a vulcanized rubber, an inorganic filler, a modified polyolefin that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups and a modified rubber that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups.","The present invention also includes a tire component including a tire tread that comprises a vulcanized rubber, an inorganic filler, a modified rubber that contains a polymerized metal salt of an unsaturated carboxylic acid, and wherein the tire component is devoid of zinc oxide.","Additionally, the present invention also provides a tire prepared by a process comprising the steps of vulcanizing a rubber composition that has been prepared by compounding ingredients comprising i) a rubber, ii) an inorganic filler, iii) a modified rubber that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups or a polymerized metal salt of an unsaturated carboxylic acid and iv) optionally a modified polyolefin that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups.","The present invention also provides a tire component including a tire tread that comprises a vulcanized rubber, an inorganic filler, and a modified rubber, where the modified rubber is prepared by polymerizing i) an unsaturated carboxylic acid, ii) an unsaturated anhydride, iii) a metal salt of an unsaturated carboxylic acid or iv) combinations of i, ii or iii, to an elastomeric backbone via free-radical polymerization.","The addition of poly(metal carboxylate) grafted rubbers to tire formulations including inorganic fillers has surprisingly been found to stabilize the dynamic modulus of the tire rubber at high temperatures.","Also, the use of these poly(metal carboxylate) grafted rubbers has surprisingly lead to an increased tan δ at higher temperature, as well as decreased tire wear in tires filled with inorganic filler.","This invention advantageously increases G′ of tire treads containing rubber fillers.","Among other advantages, tire treads containing the modified rubbers of this invention also have improved dry traction.","Another advantage of certain embodiments of this invention is the elimination of zinc oxide as an additive in tire rubber formulations."],["FIELD OF THE INVENTION This invention relates to rubber compositions and tire components that have increased dynamic moduli, as well as tire components having improved stable modulus at higher temperatures.","BACKGROUND OF THE INVENTION Hysteresis loss and storage modulus are properties of rubber that have a significant impact on tire performance.","Hysteresis loss can be defined by the formula tan δ=G″/G′, where G″ is a measure of the dynamic loss modulus and G′ is the measure of storage modulus.","Generally, as the temperature of a tire increase, G′ decreases.","As a result, there may be a decrease in handling.","Inorganic fillers, such as silica, impart improved wet traction (as demonstrated by a high tan δ at 0° C.) and improved snow traction (as demonstrated by a low tan δ at −20° C.) when used as filler within tire treads.","Mixing silica into a tire stock, however, is difficult because silica particles agglomerate and therefore they are not easily dispersed.","In response, processing and dispersing aids are used during compounding.","Unfortunately, the use of these processing and dispersing aids enhances the decrease in modulus that is typically observed at high temperatures.","Because inorganic fillers are technologically useful, there is a need to overcome the problems associated with the loss of G′ at elevated temperatures in tire treads containing inorganic fillers.","SUMMARY OF THE INVENTION In general the present invention provides a tire component including a tire tread that comprises a vulcanized rubber, an inorganic filler, and a modified rubber, where the modified rubber i) contains pendent or functional groups that contain carboxylic acid or anhydride groups, ii) has pendently grafted thereto a polymerized metal salt of an unsaturated carboxylic acid, or iii) contains both i and ii.","The present invention also includes a tire tread comprising a vulcanized rubber, from about 10 to about 100 parts by weight inorganic filler per 100 parts by weight rubber, and from about 0.2 to about 10 parts by weight modified rubber per 100 parts by weight rubber.","The present invention also provides a tire component including a tire tread that comprises a vulcanized rubber, an inorganic filler, a modified polyolefin that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups and a modified rubber that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups.","The present invention also includes a tire component including a tire tread that comprises a vulcanized rubber, an inorganic filler, a modified rubber that contains a polymerized metal salt of an unsaturated carboxylic acid, and wherein the tire component is devoid of zinc oxide.","Additionally, the present invention also provides a tire prepared by a process comprising the steps of vulcanizing a rubber composition that has been prepared by compounding ingredients comprising i) a rubber, ii) an inorganic filler, iii) a modified rubber that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups or a polymerized metal salt of an unsaturated carboxylic acid and iv) optionally a modified polyolefin that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups.","The present invention also provides a tire component including a tire tread that comprises a vulcanized rubber, an inorganic filler, and a modified rubber, where the modified rubber is prepared by polymerizing i) an unsaturated carboxylic acid, ii) an unsaturated anhydride, iii) a metal salt of an unsaturated carboxylic acid or iv) combinations of i, ii or iii, to an elastomeric backbone via free-radical polymerization.","The addition of poly(metal carboxylate) grafted rubbers to tire formulations including inorganic fillers has surprisingly been found to stabilize the dynamic modulus of the tire rubber at high temperatures.","Also, the use of these poly(metal carboxylate) grafted rubbers has surprisingly lead to an increased tan δ at higher temperature, as well as decreased tire wear in tires filled with inorganic filler.","This invention advantageously increases G′ of tire treads containing rubber fillers.","Among other advantages, tire treads containing the modified rubbers of this invention also have improved dry traction.","Another advantage of certain embodiments of this invention is the elimination of zinc oxide as an additive in tire rubber formulations.","BRIEF DESCRIPTION OF THE DRAWINGS FIG.","1 is a graphical plot of the temperature dependence of G′ of a vulcanizate prepared according to the present invention as compared to a control.","FIG.","2 is a graphical plot of the strain dependence of tan δ of a vulcanizate prepared according to the present invention as compared to a control.","FIG.","3 is a graphical plot of the strain dependence of G′ of a vulcanizate according to the present invention as compared to controls, where one control employs carbon black as a filler.","DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS Tires and tire components are prepared from rubber compositions that include a) a rubber, b) a modified rubber, wherein said modified rubber i) contains pendent or functional groups that contain carboxylic acid or anhydride groups, ii) contains pendently grafted thereto a polymerized metal salt of an unsaturated carboxylic acid, or iii) both i and ii, c) a inorganic filler, and d) an optional modified polyolefin that contains pendent or terminal functional groups that contain carboxylic acid or anhydride groups.","Other additives that are typically employed in rubber compositions, especially those used to fabricate tires, may also be included.","Both synthetic and natural rubber may be employed within the rubber compositions.","These rubbers, which may also be referred to as elastomers, include, without limitation, natural rubber, synthetic poly(isoprene), poly(styrene-co-butadiene), poly(butadiene), poly(styrene-co-butadiene-co-isoprene), poly(styrene-co-isoprene), and mixtures thereof.","In the first embodiment, the modified rubber additive is an elastomeric polymer that contains terminal or pendent functional groups that contain carboxylic acid or anhydride groups.","The terminal or pendent functional groups that contain the carboxylic acid or anhydride groups may be obtained by grafting functional groups to a polymeric chain or by preparing a copolymer by using at least one monomer that will yield the desired functional group The modified rubber additive can be obtained by polymerizing unsaturated carboxylic acids or unsaturated anhydrides from a graft point on an elastomeric polymer.","For example, functional groups may be obtained by the free radical polymerization of methacrylic acid in the presence of the diene rubber.","Examples of elastomers from which this grafting reaction may take place include poly(butadiene), poly(styreneco-butadiene), poly(isoprene), poly(styrene-co-butadiene-co-isoprene), poly(styrene-co-isoprene), and terpolymers of ethylene, an a-olefin, and diene monomers.","Examples of saturated carboxylic acids that can undergo polymerization and graft to an elastomer include citraconic acid, cinnamic acid, methacrylic acid, itaconic acid, and acrylic acid.","Examples of unsaturated anhydrides that can undergo polymerization and graft to an elastomer include maleic anhydride, citraconic anhydride and itaconic anhydride.","Free radical polymerization is the preferred reaction for grafting these monomers to an elastomer.","Preferably, this technique employs an initiator such as a thermo-decomposition initiator.","Examples of these initiators include but are not limited to, di-sec-butyl peroxydicarbonate, t-amyl peroxy pivalate, 2,5-dimethyl-2,5-di-(2-ethylhexanoyl-peroxy) hexane, t-amylperoxy-2-ethylhexanoate, t-butyl-2-ethylhexanoate, 2,2-azo-bis-(2-methyl propionitrile), azo-bis-isobutyronitrile (AIBN) and the like.","This grafting reaction preferably takes place in an inert solvent such as a hydrocarbon solvent.","Where the functional group is grafted to an elastomeric polymer to form the modified rubber additive, the resulting grafted copolymer may contain from about 1 to about 50 percent by weight, preferably from about 2 to about 35 percent by weight, and even more preferably from about 3 to about 25 percent by weight of the grafted functional groups.","Alternatively, the modified rubber can be obtained by copolymerizing unsaturated carboxylic acid monomers or unsaturated anhydride monomers with diene or other monomers that will provide a rubbery polymer.","Examples of unsaturated carboxylic acids and unsaturated anhydrides are provided above.","Examples of diene monomers that will yield a rubbery polymer include diene monomers such as 1,3-butadiene, and isoprene.","Other monomer that may be copolymerized with these diene monomers include vinyl aromatic monomer such as styrene.","This copolymerization technique is well-known, and it typically takes place in an emulsion polymerization with the use of a radical source such as a peroxide redox system.","The number average molecular weight of the modified rubber additive of this embodiment can vary greatly, although it is preferred that the number average molecular weight (Mn) be from about 200,000 to about 500,000.The modified rubber additive of the first embodiment generally contains from about 0.02 to about 28 milliequivalents of functional groups (e.g., units deriving from methacrylic acid) per gram of modified polymer, preferably from about 0.1 to about 25, and more preferably from about 0.5 to about 20 milliquivalents of functional groups per gram of modified polymer.","The second embodiment of the present invention is the poly(metal carboxylate) graft rubber, which is a macromolecular structure that comprises an unsaturated hydrocarbon rubber backbone having pendently grafted thereto a polymerized metal salt of an unsaturated carboxylic acid.","Each macromolecular structure will contain at least one pendent graft, preferably at least 2 pendent grafts, and more preferably at least 5 pendent grafts per mole of rubber polymer chain.","This grafted rubber is described in U.S. Pat.","No.","5,962,593, which is incorporated herein by reference.","Any unsaturated hydrocarbon rubber may be utilized to form the polymeric backbone of the graft copolymer rubber.","Preferred unsaturated hydrocarbon rubbers include homopolymers of conjugated dienes and copolymers of conjugated dienes and other monomers such as vinyl aromatics.","Exemplary conjugated diene monomers include 1,3-butadiene, isoprene, and 2,3-dimethyl-1,3-butadiene.","Exemplary vinyl aromatic monomers include styrene, α-methylstyrene, and p-methylstyrene.","Other preferred hydrocarbon rubbers include terpolymers of ethylene, propylene, and dienes such as, but not limited to, 5-ethylidene-2-norbornene and 1,4-hexadiene.","Some exemplary polymers include poly(butadiene), poly(styrene-co-butadiene), poly(ethylene-co-propylene-co-diene), poly(isoprene), poly(styrene-co-isoprene), and poly(butadiene-co-styrene-co-isoprene).","Any unsaturated carboxylic acid may be used to form the metal salt.","The unsaturated carboxylic acids include α,β-ethylenically unsaturated carboxylic acids having from 3 to 8 carbon atoms.","Exemplary carboxylic acids include acrylic, methacrylic, cinnamic, and crotonic acids, with acrylic and methacrylic acids being preferred.","Suitable metal ions that may be used to form the metal salts include sodium, potassium, magnesium, calcium, zinc, barium, aluminum, tin, zirconium, lithium and cadmium, with zinc and magnesium being preferred.","The metal ion is preferably introduced in the form of a salt of the carboxylic acid.","A preferred metal salt is zinc dimethacrylate.","The poly(metal carboxylate) graft rubber can be prepared by (i) dissolving an unsaturated hydrocarbon rubber in a solvent, (ii) adding to the dissolved rubber a metal salt of an unsaturated carboxylic acid to form a rubber/carboxylic acid blend, (iii) adding to the rubber/carboxylic acid blend a free radical initiator, and (iv) heating the reaction mixture to a temperature of from about 40 to about 150° C. for about 0.1 to about 100 hours.","Preferably, the reactor containing the rubber/carboxylic acid blend is sealed prior to adding the initiator.","Once sealed, the initiator is preferably added in liquid form under pressure.","The metal salt is preferably added as a fine powder with stirring and under a nitrogen atmosphere to form a dispersion of the metal salt in the polymer solution.","Preferably, a surfactant is added along with the metal salt in order to provide a more stable dispersion of the metal salt in the polymer solution.","Preferred surfactants include nonionic octylphenoxy polyethoxy ethanol surfactants, which available as Triton X-15, Triton X-45, and Triton X-100 (Rohm and Haas Co.; Philadelphia, Pa.).","Preferably, any solvent in which both the rubber and the free radical initiator are soluble may be used.","The preferred solvents are the hydrocarbon solvents in which the original polymers were prepared.","Exemplary solvents are aromatic and aliphatic hydrocarbons including, but not limited to, hexane, heptane, pentane, octane, cyclohexane, cycloheptane, cyclopentane, methyl cyclohexane, benzene, and toluene.","Hexane and toluene are preferred.","Suitable free radical initiators include, but are not limited to, di-sec-butyl peroxydicarbonate, t-amyl peroxy pivalate, 2,5-dimethyl-2,5-di-(2-ethylhexanoyl-peroxy) hexane, t-amylperoxy-2-ethylhexanoate, t-butyl-2-ethylhexanoate, 2,2-azo-bis-(2-methyl propionitrile), azo-bis-isobutyronitrile (AIBN) and the like.","The optional modified polyolefin additive includes an α-olefin polymer that contains terminal or pendent moieties containing acid or anhydride groups.","The terminal or pendent moieties typically derive from unsaturated carboxylic acids or unsaturated anhydrides.","Examples of unsaturated carboxylic acids include citraconic acid, cinnamic acid, methacrylic acid, and itaconic acid.","Examples of unsaturated anhydrides include maleic anhydride, citraconic anhydride, and itaconic anhydride.","The preferred terminal or pendent moieties are succinic anhydride groups, or the corresponding acid from a ring opening structure, that derives from maleic anhydride.","The α-olefin polymer includes an α-olefin homopolymer, a copolymer of two or more α-olefins, or a copolymer of an α-olefin with a compatible monomer.","The α-olefins can include from about 2 to about 8 carbon atoms, and more preferably from 3 to about 5 carbon atoms.","Exemplary α-olefin monomers include ethylene, propylene, butene-1, and pentene-1.Exemplary monomers that can be copolymerized with α-olefins include vinyl aromatic monomers and diene monomers.","An exemplary copolymer is poly(propylene-co-ethylene) that contains polyethylene crystals.","The modified polyolefins should contain from about 0.01 to about 3 pbw of the functional moiety based upon the weight of the entire polymer.","More preferably, the modified polyolefin should contain from about 0.1 to about 2 pbw of the functional moiety, an even more preferably from about 0.15 to about 1.0 pbw of the functional moiety.","The number average molecular weight of the modified polyolefins can vary greatly, although it is preferred that the number average molecular weight (Mn) be from about 20,000 to about 500,000, more preferably from about 100,000 to about 400,000, and even more preferably from about 150,000 to about 400,000, as determined by using standard GPC analysis with polystyrene as a standard.","Generally, the molecular weight distribution (Mw/Mn) should be less than about 4.5, preferably less than about 4.0, and even more preferably less than about 3.8.The modified polyolefin additives are typically prepared by grafting unsaturated carboxylic acids or unsaturated anhydrides to a polyolefin polymer.","The techniques employed to attach the terminal or pendent moieties that contain carboxylic acid or anhydride groups to a polyolefin polymer are well known in the art.","For example, grafting maleic anhydride to a polyolefin is disclosed in U. S. Pat.","No.","6,046,279, which is incorporated herein by reference.","The α-olefin polymers can be synthesized by using a number of polymerization techniques such as, but not limited to, the “Phillips catalyzed reactions” conventional Ziegler-Natta type polymerizations, and metallocene catalysis including, but not limited to, metallocene-aluminoxane and metallocene-ionic activator catalysis.","Exemplary α-olefin polymers include polyethylene, polypropylene, poly(ethylene-co-propylene), poly(propylene-co-butene-1), and poly(butene-1-co-ethylene).","These α-olefin polymers can be either amorphous, semi-crystalline, or crystalline polymers.","The preferred polyolefins include crystalline or stereoregular polypropylene.","Most polypropylene homopolymers that are commercially produced have an isotactic microstructure.","The poly(propylene-co-ethylene) copolymers can be random or block copolymers.","Preferably, these copolymers will contain some polyethylene crystals, although they should include a major amount of propylene units and only a minor amount of ethylene units.","Preferably, these copolymers should contain less than about 40 percent by weight (pbw) ethylene units, more preferably from about 1 to about 30 pbw ethylene units, and more preferably from about 1.5 to about 25 pbw ethylene units.","Modified polyolefins are commercially available.","For example, maleic anhydride functionalized polypropylene is available under the tradename EXXELOR™ P01015 & 1020 (Exxon Mobil Chemical Company; Houston, Tex.","), under the tradename PP-C™, CA1000, or 18707 (Elf Atochem; Philadelphia, Pa.), or under the tradename Polybond™ 3001, 3002, or 3150 (Uniroyal Chemical Company; Middlebury, Conn.).","The rubber compositions preferably include an inorganic filler as well as optional organic fillers.","The inorganic fillers may include silica, aluminum hydroxide, magnesium hydroxide, and clays (hydrated aluminum silicates).","The optional organic fillers includes carbon black.","Useful silica (silicon dioxide) includes wet-process, hydrated silica produced by a chemical reaction in water, and precipitated as ultrafine spherical particles.","These particles strongly associate into aggregates that in turn combine less strongly into agglomerates.","The surface area, as measured by the BET method, gives the best measure of the reinforcing character of different silicas.","Useful silicas preferably have a surface area of about 32 to about 400 m2/g, preferably about 100 to about 250 m2/g, and more preferably about 150 to about 220 m2/g.","The pH of the silica filler is generally about 5.5 to about 7 and preferably about 5.5 to about 6.8.Commercially available silicas include Hi-Sil™ 215, Hi-Sil™ 233, and Hi-Sil™ 190 (PPG Industries; Pittsburgh, Pa.).","Useful commercial grades of different silicas are also available from other sources including Rhone Poulenc.","Useful aluminum hydroxide fillers include any aluminum hydroxide filler including those that have conventionally been employed to reinforce rubber compositions and tire components.","Aluminum hydroxide filler preferably has an average diameter of about 20 nanometers (nm) to about 2,000 nm, more preferably from about 25 nm to about 1,000 nm or smaller, and even more preferably from about 30 nm to about 50 nm.","Useful aluminum hydroxide preferably has a BET specific surface area of from about 0.5 to about 500 m2/g, more preferably from about 1 to about 400 m2/g, and even more preferably from about 2 to about 300 m2/g.","Aluminum hydroxide filler may be treated with various surface treating agents.","Non-limiting examples of surface treating agents include organic treating agents such as fatty acids and alcohols, and inorganic dispersants such as coupling agents and fatty acid metal salts.","Useful aluminum hydroxide filler is commercially available from a number of sources including that available under the tradenames Hygilite™ (Showa Dendo K.K.","; Japan), Baikalox™(Baikowski Chimie; France), and RF22™ (Sumitono Chemical Co.; Japan).","Useful carbon black may include any commonly available carbon black, but those having a surface area (EMSA) of at least 20 m2/g, and more preferably at least 35 m2/g up to 200 m2/g or higher, are preferred.","Surface area values used in this application are those determined by ASTM test D-1765 by using the cetyltrimethyl-amnmonium bromide (CTAB) technique.","A coupling agent may optionally be added when an inorganic filler such as silica is used.","Coupling agents include those compounds that are believed to react with both the rubber and the inorganic filler.","One coupling agent conventionally used is bis-[3(triethoxysilyl) propyl]-tetrasulfide, which is commercially available under the tradename S169 (Degussa, Inc.; New York, N.Y.).","Other coupling agents include bis-[3(tdethoxysilyl) propyl]-disulfide, which is commercially available under the tradename Silquest™ (Witco; Greenwich, Conn.), mercapto propyl alkoxy silane, which is commercially available under the tradename Ciptane™ (Dow Corning; Midland, Mich.), and dithio diproprionic acid or carboxylic acid disulfides, which are commercially available from Aldrich Chemical Company.","In general, these coupling agents should be used in an amount from about 0.1 to 20% by weight based upon the weight of the inorganic filler.","Shielding and dispersing agents, which prevent or alleviate the agglomeration of inorganic filler particles such as silica, may also be used.","Typically, these agents react or interact with the filler.","Exemplary dispersing or shielding agents include silanes, amines, diols, polyethers, amides, and sugar fatty acid esters.","U.S. Pat.","Nos.","5,719,207, 5,780,538, 5,717,022, and EP 0890606 are incorporated herein by reference in this regard.","Specific examples of these agents include sugar, fatty acids such as sorbitan fatty acids which are available from BASF (Mount Olive, N.J.), and octyl triethoxy silane, which is available from Dow Corning (Midland, Mich.).","Generally, these shielding or dispersing agents may be used in an amount from about 0.1 to about 20% by weight based on the weight of the inorganic filler.","In preferred embodiments, the coupling agents, shielding agents, and dispersing agents may be used in combination.","The rubber compositions of this invention can be cured in a conventional manner with known vulcanizing agents.","For example, sulfur or peroxide-based curing systems may be employed.","Also, see Kirk-Othmer, ENCYCLOPEDIA OF CHEMICAL TECHNOLOGY, 3rd Edition, Wiley Interscience, N.Y. 1982, Vol.","20, pp.","365-468, particularly VULCANIZATION AGENTS AND AUXILIARY MATERIALS pp.","390-402, or Vulcanization by A. Y. Coran, ENCYCLOPEDIA OF POLYMER SCIENCE AND ENGINEERING, 2nd Edition, John Wiley & Sons, Inc., 1989, which are incorporated herein by reference.","Vulcanizing agents may be used alone or in combination.","This invention does not appreciably affect cure times.","Typically, vulcanization is effected by heating the vulcanizable composition; e.g., it is heated to about 170° C.. Cured or crosslinked polymers may be referred to as vulcanizates, which are generally three-dimensional polymeric networks that are thermoset.","Depending on the degree of unsaturation of the modified rubber, it may become part of the crosslinked network comprising the vulcanizate.","Alternatively, if the modified rubber is highly saturated, it will be dispersed within the crosslinked polymeric network.","The other ingredients, such as the modified polyolefin and fillers, are generally dispersed throughout this thermoset network.","The rubber compositions and tire components may also include other compounding additives such as accelerators, oils, waxes, scorch inhibiting agents, and processing aids.","Compositions containing synthetic rubbers typically include antidegradants, processing oils, zinc oxide, optional tackifying resins, optional reinforcing resins, optional fatty acids, optional peptizers, and optional scorch inhibiting agents.","The use of the poly(metal carboxylate) graft rubber according to this invention can, advantageously, eliminate the need for ingredients such as zinc oxide and stearic acid.","The use of the terminal or pendent functional groups that contain the carboxylic acid or anhydride groups with the modified polyolefin can also eliminate the need for ingredients such as zinc oxide.","Accordingly, the rubber compositions of certain embodiments of this invention may be essentially devoid of zinc oxide and stearic acid, where essentially devoid refers to the absence of any amount that is conventionally required to impact compounding and curing.","The rubber compositions and tire components of the present invention will generally contain from about 0.1 to about 25 parts by weight of the modified rubber additive per 100 parts by weight rubber (phr).","Preferably, the rubber compositions and tire components will contain from about 0.2 to about 20 parts by weight phr, even more preferably from about 0.5 to about 15 parts by weight phr, and still more preferably from about 1 to about 10 parts by weight phr of the modified rubber additive.","The rubber compositions and tire components of the present invention will optionally contain from about 0.1 to about 25 parts by weight of the modified polyolefin additive per 100 parts by weight rubber (phr).","Preferably, the rubber compositions and tire components will contain from about 0.2 to about 20 parts by weight phr, even more preferably from about 0.5 to about 15 parts by weight phr, and still more preferably from about 1 to about 10 parts by weight phr of the modified polyolefin additive.","Fillers, such as carbon black, silica or aluminum hydroxide, are typically employed in an amount from about 1 to about 100 parts by weight phr, and preferably from about 20 to about 90 parts by weight phr, and more preferably from about 40 to about 80 parts by weight phr.","Silica may optionally be used in an amount from about 10 to about 100 parts by weight phr, preferably from about 15 to about 90 parts by weight phr, and more preferably from about 20 to about 80 parts by weight phr.","Carbon black may optionally be used in an amount from about 0.5 to about 80 parts by weight phr, preferably from about 1 to about 40 parts by weight phr, and more preferably from about 2 to about 30 parts by weight phr.","Optionally, aluminum hydroxide filler should be employed in an amount from about 1 to about 25 parts by weight phr, preferably from about 2 to about 20 parts by weight phr, and more preferably from about 5 to about 15 parts by weight phr.","In a preferred embodiment, aluminum hydroxide, silica, and carbon black are used in combination.","In this embodiment, the silica:carbon black ratio is from about 1:10 to about 99:1, more preferably from about 1:5 to about 5:1, and even more preferably from about 1:3 to about 3:1.And, the silica:aluminum hydroxide ratio is from about 3:1 to about 30:1, more preferably from about 5:1 to about 20:1, and even more preferably from about 6:1 to about 15:1.Those skilled in the art will be able to choose a useful amount of the other ingredients that may be employed in practicing the present invention.","For example, it is generally known in the art of making tire components, such as treads, that sulfur is typically employed in an amount from about 0.5 to about 10 parts by weight phr, and preferably from about 1 to about 6 parts by weight phr.","Oils are typically employed in an amount from about 1 to about 60 parts by weight phr, and preferably in an amount from about 1 to about 50 parts by weight phr.","Zinc oxide is typically employed in an amount from about 0.5 to about 8, and preferably from about 1 to about 5 parts by weight phr, however, compositions which contain the poly(metal carboxylate) graft rubber according to this invention, or modified rubber having terminal or pendent functional groups that contain the carboxylic acid or anhydride groups and the modified polyolefin of the present invention may eliminate the need for ingredients such as zinc oxide and stearic acid.","Tire formulations are compounded by using mixing equipment and procedures conventionally employed in the art.","Preferably, an initial masterbatch is prepared that includes the elastomer component and the reinforcing fillers, as well as other optional additives such as processing oil and antioxidants.","The modified rubbers are preferably added during preparation of the initial masterbatch.","Once this initial masterbatch is prepared, the vulcanizing agents are blended into the composition.","Rubber compounding techniques and the additives employed therein are generally known as disclosed in The Compounding and Vulcanization of Rubber, by Stevens in RUBBER TECHNOLOGY SECOND EDITION (1973 Van Nostrand Reihold Company).","The mixing conditions and procedures applicable to silica-filled tire formulations are also well known as described in U.S. Pat.","Nos.","5,227,425, 5,719,207, 5,717,022, as well as EP 0890606, all of which are incorporated herein by reference.","The composition can then be processed into tire components according to ordinary tire manufacturing techniques including standard rubber curing techniques.","Tire components of this invention preferably include tire treads.","The compositions, however, can also be used to form other elastomeric tire components such as subtreads, black sidewalls, body ply skims, bead fillers and the like.","Pneumatic tires can be made according to U.S. Pat.","Nos.","5,866,171; 5,876,527; 5,931,211; and 5,971,046, which are incorporated herein by reference.","Tire components of this invention preferably include tire treads.","The composition can also be used to form other elastomeric tire components such as subtreads, sidewalls, body ply skims, bead fillers and the like.","The use of polyolefin additives within tires treads that contain the modified rubbers of the present invention leads to both reduced hysteresis loss while improving handling performance without the need for other additives such as particulate nylons.","In order to demonstrate the practice of the present invention, the following examples have been prepared and tested as described in the General Experimentation Section disclosed hereinbelow.","The examples should not, however, be viewed as limiting the scope of the invention.","The claims will serve to define the invention.","EXAMPLES Example 1 Stocks 1-2 Two tire stocks were prepared from the formulations in Table I by using the compounding protocol set forth in Table II within a 310 gram Brabender mixer set at 60 r.p.m.","TABLE I Stock 1 Stock 2 Polybutadiene 100 90 Poly(butadiene-g-methacrylic acid) 0 10 Precipitated silica 65.00 65.00 Aromatic Process oil 16.25 16.25 Wax 1.5 1.5 Antioxidant [N-(1,3 dimethylbutyl)-N′-phenyl-p- 1.70 1.70 phenylene-diamine] Si69 6.5 6.5 Sulfur 2.85 2.85 Steric acid 2.00 2.00 Accelerator [N-Cyclohexyl-2-benzothiazyl- 1.50 1.50 sulfenamide] Diphenyl Guanidine 0.50 0.50 Zinc oxide 2.5 2.5 The polybutadiene was obtained under the tradename HD55™ (Firestone Polymers; Akron, Ohio).","The poly(butadiene-g-methacrylic acid) was prepared by grafting unsaturated methacrylic acid to polybutadiene by using azobisisobutyronitrile (AIBN) as an initiator.","This grafting took place in a hydrocarbon solvent at 60-80° C. with 10 parts by weight methacrylic acid per 100 parts by weight polybutadiene.","The polybutadiene had a Mooney Viscosity (ML1+4 at 100° C.) of 52 and a cis microstructure of about 33 weight percent.","The poly(butadiene-g-methacrylic acid) had a Mooney Viscosity (ML1+4 at 100° C.) of 56, a cis microstructure of about 33 weight percent, and contained about 10 weight percent units deriving from the methacrylic acid.","TABLE II Master Batch Stage Initial Temperature 100° C. 0 sec charging polymers 30 sec charging silica and all pigments 5 min drop Drop Temperature 155° C. Remill 1 Batch Stage Initial Temperature 70° C. 0 sec charging remilled stock 30 sec charging Si69 Drop Temperature 155° C. Final Batch Stage Initial Temperature 90° C. 0 sec charging remilled stock 30 sec charging curing agent and accelerators Drop Temperature 105° C. The final stocks were sheeted and then subsequently molded at 171° C. for 15 minutes.","The Mooney Viscosity, δ(ΔG′), and curing characteristics of the uncured stocks were evaluated.","Cured samples of each stock were also evaluated for tensile mechanical properties at 25° C. Further, the dynamic viscoelastic properties of cured samples of each stock were also evaluated.","The results of this analysis are set forth in Table II.","TABLE III Stock No.","1 2 Uncured Analysis Mooney Viscosity (ML1+4 @ 100° C.) 80.8 81.7 T5 Scorch @ 130° C. (sec) 951 1,037 tS2 @ 171° C. (min) 1.07 1.18 T90 @ 171° C. (min) 6.82 6.77 δ(ΔG′) (kPa) 2,062 1,106 Tensile Properties Modulus @ 50% (MPa)) 1.63 1.79 Modulus @ 300% (MPa) 9.86 10.74 Tensile Strength @ Break (MPa) 10.90 12.64 Elongation @ Break (%) 314 333 Toughness (MPa) 14.82 17.97 Viscoelastic Properties Tanδ @ 0° C. (temperature sweep) 0.1733 0.1612 Tanδ @ 50° C. (temperature sweep) 0.1245 0.1131 ΔG′ (G′@.25%-G′@14.75%) 0.253 0.2414 @ 65° C. (strain sweep) (MPa) Tanδ @ 7% strain and 65° C. 0.1055 0.1026 (strain sweep) British Portable Skid Tester 42 42 Abrasion Resistance (%) 100 111 The Mooney viscosity measurement was conducted at 130° C. by using a larger rotor, and was recorded as the torque when the rotor has rotated for 4 minutes.","Each sample was preheated at 130° C. for one minute before the rotor started.","T5 is the time required to increase 5 Mooney units during the Mooney-scorch measurement.","This index is used to predict how fast the compound viscosity will rise during processing.","tS2 and T90 are the times required for torque rises to 2% and 90% of the total torque increase during curing characterization experiments at 171° C. These measurements are typically used to predict how fast the viscosity build-up (tS2) and the curing rate during the curing process.","The G′ of the uncured stocks was obtained from a strain sweep measurement by using an RPA2000 Rubber Process Analyzer (Alpha Technologies).","These strain sweep experiments were conducted at 50° C. at a frequency of 0.1 Hz with strain sweeping from 0.25% to 1,000%.","The difference in G′ at strains of 0.25% and 1,000% were recorded as the ΔG′.","ΔG′ was also determined after the stocks were annealed at 171° C. for 15 minutes.","δ(ΔG′), therefore, is the difference between ΔG′ before annealing and ΔG′ after annealing (ΔG′ after heat treatment−ΔG′ before heat treatment).","The tensile mechanical properties were measured by using the standard procedure described in ASTM-D 412 at 25° C. The tensile test specimens were round rings with a dimension of 1.27 mm in width and 1.905 mm in thickness.","A specific gauge length of 25.4 mm was used for the tensile test.","Temperature sweep experiments were conducted with a frequency of 31.4 rad/sec by using 0.5% stain for temperatures ranging from −100° C. to −10° C., and 2% strain for temperatures ranging from −10° C. to 100° C. Payne effect (ΔG′) and tanδ at 7% strain were obtained from the strain sweep experiments.","A frequency of 3.14 rad/sec was used for strain sweep experiments conducted at 65° C. with strain sweeps from 0.25% to 14.75%.","The wet traction of the rubber was evaluated by using the British Portable Skid Tester.","The details of this test are described in ASTM E-303, Vol.","04.03.According to this test, a specimen is attached to the base of a pendulum arm, and the specimen contacts the opposing surface during a swing of the pendulum.","The weighted pendulum head is free to move vertically on the pendulum arm so that the swing amplitude determines the friction of the rubber against the pavement surface.","The lower the amplitude that the pendulum swings up after contacting the surface (recorded as a higher value on the scale of the tester), the higher friction of the rubber against the surface.","The wear resistance of the cured samples were evaluated by weighing the amount of wear according to the Lambourne test.","The wearing index was obtained from the ratio of the weight loss of the control to that of the testing sample.","Samples with higher wear indices have better wear resistance properties.","Samples used for this Lambourne test are circular donuts with the following approximate dimensions: 22.86 mm and 48.26 mm for the inside and outside diameters, and 4.95 mm in thickness.","The test specimens were place on an axil and run at a slip ratio of 25% against a driven abrasive surface.","Stocks 3-6 Four tire stocks were prepared from the formulations in Table IV by using the compounding protocol set forth in Table V within a 310 gram Brabender mixer set at 60 r.p.m.","TABLE IV Stock 3 Stock 4 Stock 5 Stock 6 Polybutadiene 90 90 100 90 poly(butadiene-g-methacrylic 10 10 0 10 acid) Polypropylene 0 10 0 00 poly((propylene-co-ethylene)- 0 0 10 10 g-maleic anhydride) Precipitated silica 65 65 65 65 Aromatic Process oil 16.25 16.25 16.25 16.25 Wax 1.5 1.5 1.5 1.5 Antioxidant [N- 0.95 0.95 0.95 0.95 (1,3 dimethylbutyl)- N′-phenyl-p-phenylene- diamine] Si69 6.50 6.50 6.50 6.50 Sulfur 2.85 2.85 2.85 2.85 Steric acid 1.50 1.50 1.50 1.50 Accelerator [N-Cyclohexyl-2- 1.50 1.50 1.50 1.50 benzothiazyl-sulfenamide] Diphenyl Guanidine 0.5 0.5 0.5 0.5 Zinc oxide 2.5 2.5 2.5 2.5 The polybutadiene and poly(butadiene-g-methacrylic acid) were the same as those from Table I.","This grafting took place under substantially the same conditions as used for Samples 1 and 2.The polypropylene was obtained under the tradename PP340™ (Aldrich) and was characterized by having a weight average molecular weight (Mw) of about 340,000, a melt temperature (Tm) of about 161° C., and a crystallinity of greater than about 75%.","The poly((propylene-co-ethylene)-g-maleic acid) was obtained under the tradename Orevac™ 18707 (Elf Atochem) and was characterized by having a Mw of about 360,000, a Tm of about 120° C. and about 150° C., a crystallinity that was greater than about 75%, an ethylene content of about 21 weight percent, and contained about 0.48 weight percent of moieties deriving from maleic anhydride.","TABLE V Master Batch Stage Initial Temperature 100° C. 0 sec charging polymers and polyolefin (if added) 30 sec charging silica and all pigments 5 min drop Drop Temperature 175° C. Remill 1 Batch Stage Initial Temperature 70° C. 0 sec charging remilled stock charging rest of the silica 30 sec charging Si69 Drop Temperature 155° C. Final Batch Stage Initial Temperature 90° C. 0 sec charging remilled stock 30 sec charging curing agent and accelerators Drop Temperature 105° C. The final stocks were sheeted and then subsequently molded at 171° C. for 15 minutes.","The Mooney Viscosity, δ(ΔG′), and curing characteristics of the uncured stocks were evaluated.","Cured samples of each stock were also evaluated for tensile mechanical properties at 25° C. and after heat aging at 100° C. for 24 hours.","Further, the dynamic viscoelastic properties of cured samples of each stock were also evaluated.","The results of this analysis are set forth in Table VI.","TABLE VI Stock No.","3 4 5 6 Uncured Analysis Mooney Viscosity (ML1+4 @ 100° C.) 81.7 110.7 98.4 101.1 T5 Scorch @ 130° C. (min) 1,037 874 926 930 tS2 @ 171° C. (min) 1.18 1.37 1.27 1.23 T90 @ 171° C. (min) 6.77 6.47 6.79 6.90 Δ(ΔG′) (kPa) 1,106 3,224 1,622 987 Tensile Properties (ring-shaped samples) Modulus @ 50% (MPa) 1.79 2.56 2.10 2.12 Modulus @ 300% (MPa) 10.74 13.56 11.09 11.16 Tensile Strength @ Break (MPa) 12.64 16.55 14.24 16.02 Elongation @ Break (%) 333 353 361 389 Toughness (MPa) 17.97 27.54 23.74 27.98 Tensile Properties (dumbbell-shaped samples) Modulus @ 50% (MPa) 1.79 3.08 2.37 2.33 Modulus @ 300% (MPa) 7.52 11.75 9.43 9.25 Tensile Strength @ Break (MPa) 13.73 16.35 14.15 16.28 Elongation @ Break (%) 511 432 448 518 Toughness (MPa) 33.90 38.00 33.00 42.46 Viscoelastic Properties Tanδ @ 0° C. (temperature sweep) 0.1612 0.1754 0.1744 0.1689 Tanδ @ 50° C. (temperature sweep) 0.1131 0.1255 0.1206 0.1217 ΔG′ (G′@.25%-G′@14.75%) @ 65° C. (strain sweep) (MPa) 0.79 1.37 0.98 1.085 Tanδ @ 7% strain and 65° C. (strain sweep) 0.2414 0.476 0.3813 0.3762 British Portable Skid Tester 0.1026 0.1115 0.1106 0.1120 Abrasion Resistance (%) 100 100 109 110.4 TABLE VII Stock No.","3 4 5 6 Tensile Properties @ 25° C. (after thermal aging) Modulus @ 50% (MPa) 2.37 3.61 2.85 2.79 Tensile Strength @ Break 14.15 16.86 15.79 17.24 (MPa) Elongation @ Break (%) 249 241 258 286 Toughness (MPa) 15.34 19.71 18.24 22.21 Tear Properties @ 100° C. Strength (MPa) 1.84 1.70 1.98 2.22 Elongation @ Break (%) 284 192 290 320 Stocks 7-8 Two tire stocks were prepared from the formulations of Table VIII (expressed in parts by weight) by using the compounding protocol set forth in Table II within a 310 gram Brabender mixer set at 60 r.p.m.","TABLE VIII Stock 7 Stock 8 Natural Rubber 20.0 20.0 Solution SBR 50.0 50.0 Poly(metal carboxylate) grafted rubber 0.0 5.0 Oil extended solution (poly(styrene-co- 27.5 27.5 butadiene) Poly(butadiene) 10.0 10.0 Carbon Black 33.3 33.3 Precipitated silica 33.0 33.0 Wax 1.0 1.0 Antioxidant [N-(1,3 dimethylbutyl)-N′-phenyl- 0.95 0.95 p-phenylene-diamine Sulfur 2.7 2.7 Accelerator [N-Cyclohexyl-2- 0.5 0.5 benzothiazolsulfenamine] Zinc oxide 1.5 1.5 Diphenyl Guanidine 0.5 0.5 Dioctyl phthalate 6.0 6.0 Aromatic processing oil 5.0 5.0 N-octyl triethoxy silane 1.8 1.8 Sorbitan Monooleate 5.0 5.0 Si69 0.28 0.28 The solution SBR was obtained by polymerizing 1,3-butadiene monomer and styrene monomer in hexane solvent with an alkyllithium initiator.","The living polymers were then coupled with tin tetrachloride.","The resulting polymers had a Mooney Viscosity (ML1+4 at 100° C.) of about 50, a vinyl content of about 58%, and a styrene content of about 20%.","The oil extended solution poly(styrene-co-butadiene) was similarly prepared and extended in oil.","The poly(metal carboxylate) grafted rubber was prepared by grafting zinc methacrylate to a poly(butadiene) polymer that was obtained under the tradename DIENE 55™ (Firestone Synthetic Polymers; Akron, Ohio) by using an AIBN initiator.","The resulting grafted rubber had about 33% by weight poly(zinc methacrylate) graphs.","The final compounds were sheeted and cured.","Test specimens for dynamic storage moduli experiments were formed.","The dynamic storage moduli were measured by temperature sweep experiments.","These measurements were conducted with a frequency of 31.4 rad/sec by using 0.5 percent strain for temperatures ranging from −100° C. to −10° C., and 2 percent strain for the temperature ranging from −10° C. to 100° C. A frequency of 3.14 rad/sec was used for strain sweep, which was conducted at 65° C. with strain sweeping from 0.25 percent to 14.75 percent.","The temperature dependence of the G′ of stocks 1 and 2 are shown in FIG.","1.The vulcanizates of stocks 7 and 8 were also subjected to strain sweep experiments.","These experiments were conducted at a frequency of 3.14 rad/sec was used for strain sweep, which was conducted at 65° C. with strain sweeping from 0.25 percent to 14.75 percent.","From this data, tan δ was calculated and plotted over the strain sweep as shown in FIG.","2.The vulcanizates were then tested for tensile properties according to ASTM D 412 at 25° C. The tensile test specimens were round rings with a diameter of 0.127 cm and a thickness of 0.191 cm.","A specific gauge length of 2.54 cm was used.","Table III provides the results of the tests.","The vulcanizates of stocks 7 and 8 were also evaluated for wear resistance by the Lambourne test.","Samples of this test were circular donuts with the following approximate dimensions: 22.86.mm inner diameter and 48.26 mm outer diameter, and 4.95 mm thick.","Test specimens were placed on an axle and run at a slip ratio of 25% against a driven abrasive surface.","The wear indices were obtained from the ratio of the weight loss of the control to that of the tested sample.","Samples with higher wear indices have better wear resistance properties.","Table IX provides the results of these tests.","TABLE IX Stock 7 Stock 8 50% Modulus (MPa) 1.39 1.30 300% Modulus (MPa) 10.36 9.302 Tensile at Break (MPa) 16.27 18.14 Elongation at break (%) 419 485 Toughness (MPa) 29.60 39.96 Lambourne Weight loss (g) 0.1108 0.0808 Lambourne Wear Index 100 127 Stocks 9-11 Three tire stocks were prepared from the formulations in Table X by using the compounding protocol set forth in Table II within a 310 g Brabender mixer set at 60 rpm.","TABLE X Stock 9 Stock 10 Stock 11 Natural Rubber 20.0 20.0 20.0 Solution SBR with modified chain ends 50.0 50.0 50.0 Poly(metal carboxylate) grafted rubber 0.0 5.0 5.0 Oil extended solution (poly(styrene-co- 27.5 27.5 27.5 butadiene) Poly(butadiene) 10.0 10.0 10.0 Carbon Black (SAF) 33.3 33.3 66.4 Precipitated silica 33.0 33.0 0 Wax 1.0 1.0 1.0 Antioxidant [N-(1,3 dimethylbutyl)-N′- 0.95 0.95 .95 phenyl-p-phenylene-diamine Sulfur 2.7 2.7 2.7 Accelerator [N-Cyclohexyl-2- 0.5 0.5 0.5 benzothiazolsulfenamine] Zinc oxide 1.5 1.5 1.5 Diphenyl Guanidine 0.5 0.5 0.5 Dioctyl phthalate 6.0 6.0 6.0 Aromatic processing oil 5.0 5.0 5.0 Octyl triethyl silane 1.8 1.8 1.8 Sorbitan Monooleate 5.0 5.0 5.0 Si69 0.28 0.28 0.28 The ingredients set forth in Table X were essentially the same as the ingredients set forth in Table VIII.","TABLE XI Master Batch Stage 1 Initial Temperature 100° C. 0 sec charging polymers 30 sec charging carbon black, silica, shielding agent or Silanes (if added) and all pigments.","5 min.","drop Drop temperature 175-180° C. Master Batch Stage 2 Initial Temperature 70° C. 0 sec charging remilled stock 30 sec charging Si69 (if added) and remaining silica.","Drop Temperature 155° C. Test specimens for dynamic storage moduli experiments were prepared in a similar fashion to those prepared for Stocks 7 and 8, and they were likewise analyzed in a similar fashion.","The temperature dependence of G′ for Stocks 9-11 is shown in FIG.","3.Various modifications and alterations that do not depart from the scope and spirit of this invention will become apparent to those skilled in the art.","This invention is not to be duly limited to the illustrative embodiments set forth herein"]],"string":"[\n [\n \"Tire components having improved modulus\",\n \"A tire component including a tire tread that comprises a vulcanized rubber, a rubber filler, and a modified rubber, where the modified rubber i) contains pendent or functional groups that contain carboxylic acid or anhydride groups, ii) has pendently grafted thereto a polymerized metal salt of an unsaturated carboxylic acid, or iii) contains both i and ii.\",\n \"The use of such modified rubbers in tire components advantageously increases the G′ of such components.\"\n ],\n [\n \"1.A tire component including a tire tread that comprises: a vulcanized rubber, an inorganic filler; and a modified rubber that contains i) pendent or terminal functional groups containing carboxylic acid or anhydride groups or ii) a polymerized metal salt of an unsaturated carboxylic acid.\",\n \"2.A tire component including a tire tread that comprises: a vulcanized rubber, an inorganic filler; a modified polyolefin that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups; and a modified rubber that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups.\",\n \"3.The tire component of claim 2, wherein the tire component is devoid of zinc oxide.\",\n \"4.A tire component including a tire tread that comprises: a vulcanized rubber; an inorganic filler; a modified rubber that contains a polymerized metal salt of an unsaturated carboxylic acid, and wherein the tire component is devoid of zinc oxide 5.A tire tread comprising: a vulcanized rubber; from about 10 to about 100 parts by weight inorganic filler per 100 parts by weight rubber; and from about 0.2 to about 10 parts by weight of modified rubber per 100 parts by weight rubber; and wherein said modified rubber contains i) pendent or terminal functional groups containing carboxylic acid or anhydride groups or ii) a polymerized metal salt of an unsaturated carboxylic acid.\",\n \"6.A tire prepared by a process comprising the steps of: vulcanizing a rubber composition that has been prepared by compounding ingredients comprising: a rubber, an inorganic filler; a modified rubber that contains i) pendent or terminal functional groups containing carboxylic acid or anhydride groups or ii) a polymerized metal salt of an unsaturated carboxylic acid; and optionally a modified polyolefin that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups.\",\n \"7.The tire component of tread of claims 1-5 or the tire of claim 6, wherein the modified rubber is vulcanized.\",\n \"8.The tire component or tread of claims 1-5 or the tire of claim 6, where the rubber composition further comprises carbon black.\",\n \"9.The tire component or tread of claims 1-5 or tire of claim 6, where the inorganic filler comprises silica, aluminum hydroxide, or both.\",\n \"10.The tire component of claims 1 or 4, where the polymerized metal salt of an unsaturated carboxylic acid derives from an α,β-ethylenically unsaturated carboxylic acid salt having from 3 to 8 carbon atoms.\",\n \"11.The tire component of claim 10, where the unsaturated carboxylic acid salt contains sodium, potassium, magnesium, calcium, zinc, barium, aluminum, tin, zirconium, lithium, or carbonium.\",\n \"12.The tire component or tread of claims 1-3 an 5, or the tire of claim 6, where the modified rubber includes functional groups deriving from citraconic acid, cinnamic acid, methacrylic acid, itaconic acid, or mixtures thereof, and maleic anhydride, citraconic anhydride, itaconic anhydride, or mixtures thereof 13.The tire component or tread or claims 1-5 or the tire of claim 6, where the rubber composition further comprises a coupling agent, a dispersing agent, or both.\",\n \"14.The tire component or treads of claims 1, 4-5, or the tire of claim 6, where the modified rubber is a macromolecule comprising an unsaturated hydrocarbon rubber backbone having pendently grafted thereto a polymerized metal salt of an unsaturated carboxylic acid.\",\n \"15.The tire component or tread or tire of claim 14, where the unsaturated rubber backbone is a homopolymer of a conjugated diene, a copolymer of a conjugated diene and a vinyl aromatic monomer, or a terpolymer of ethylene, propylene, and a diene.\",\n \"16.The tire component or tread or tire of claim 12, where the modified rubber contains from about 0.02 to about 28 milliequivalents of functional groups per gram of modified polymer.\",\n \"17.The tire of claim 6, where the rubber composition comprises from about 1 to about 100 parts by weight inorganic filler phr, from about 0.1 to about 25 parts by weight modified rubber phr, and from about 0.1 to about 25 parts by weight of the modified polyolefin phr.\",\n \"18.The tire component or tread of claims 1-5, or the tire of claim 6, where the tire is devoid of particulate nylon.\",\n \"19.The tire component or tread or claims 1-4 or the tire of claim 6, where, the poly(metal carboxylate) graft rubber is prepared by i) dissolving an unsaturated hydrocarbon rubber in a solvent, ii) adding to the dissolved rubber a metal salt of an unsaturated carboxylic acid to form a rubber/carboxylic acid blend, iii) adding to the rubber/carboxylic acid blend a free radical initiator, and iv) heating the reaction mixture to a temperature of from about 40 to about 150° C. for about 0.1 to about 100 hours.\",\n \"20.A rubber composition comprising: a vulcanizable rubber, a filler; and a modified rubber that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups.\",\n \"21.The rubber composition of claim 20, where the rubber composition further comprises a modified polyolefin that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups.\",\n \"22.The rubber composition of claims 20 or 21, wherein the filler is carbon black.\",\n \"23.The rubber composition of claims 20 or 21, wherein the filler is a mixture of carbon black and silica.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>Hysteresis loss and storage modulus are properties of rubber that have a significant impact on tire performance.\",\n \"Hysteresis loss can be defined by the formula tan δ=G″/G′, where G″ is a measure of the dynamic loss modulus and G′ is the measure of storage modulus.\",\n \"Generally, as the temperature of a tire increase, G′ decreases.\",\n \"As a result, there may be a decrease in handling.\",\n \"Inorganic fillers, such as silica, impart improved wet traction (as demonstrated by a high tan δ at 0° C.) and improved snow traction (as demonstrated by a low tan δ at −20° C.) when used as filler within tire treads.\",\n \"Mixing silica into a tire stock, however, is difficult because silica particles agglomerate and therefore they are not easily dispersed.\",\n \"In response, processing and dispersing aids are used during compounding.\",\n \"Unfortunately, the use of these processing and dispersing aids enhances the decrease in modulus that is typically observed at high temperatures.\",\n \"Because inorganic fillers are technologically useful, there is a need to overcome the problems associated with the loss of G′ at elevated temperatures in tire treads containing inorganic fillers.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>In general the present invention provides a tire component including a tire tread that comprises a vulcanized rubber, an inorganic filler, and a modified rubber, where the modified rubber i) contains pendent or functional groups that contain carboxylic acid or anhydride groups, ii) has pendently grafted thereto a polymerized metal salt of an unsaturated carboxylic acid, or iii) contains both i and ii.\",\n \"The present invention also includes a tire tread comprising a vulcanized rubber, from about 10 to about 100 parts by weight inorganic filler per 100 parts by weight rubber, and from about 0.2 to about 10 parts by weight modified rubber per 100 parts by weight rubber.\",\n \"The present invention also provides a tire component including a tire tread that comprises a vulcanized rubber, an inorganic filler, a modified polyolefin that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups and a modified rubber that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups.\",\n \"The present invention also includes a tire component including a tire tread that comprises a vulcanized rubber, an inorganic filler, a modified rubber that contains a polymerized metal salt of an unsaturated carboxylic acid, and wherein the tire component is devoid of zinc oxide.\",\n \"Additionally, the present invention also provides a tire prepared by a process comprising the steps of vulcanizing a rubber composition that has been prepared by compounding ingredients comprising i) a rubber, ii) an inorganic filler, iii) a modified rubber that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups or a polymerized metal salt of an unsaturated carboxylic acid and iv) optionally a modified polyolefin that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups.\",\n \"The present invention also provides a tire component including a tire tread that comprises a vulcanized rubber, an inorganic filler, and a modified rubber, where the modified rubber is prepared by polymerizing i) an unsaturated carboxylic acid, ii) an unsaturated anhydride, iii) a metal salt of an unsaturated carboxylic acid or iv) combinations of i, ii or iii, to an elastomeric backbone via free-radical polymerization.\",\n \"The addition of poly(metal carboxylate) grafted rubbers to tire formulations including inorganic fillers has surprisingly been found to stabilize the dynamic modulus of the tire rubber at high temperatures.\",\n \"Also, the use of these poly(metal carboxylate) grafted rubbers has surprisingly lead to an increased tan δ at higher temperature, as well as decreased tire wear in tires filled with inorganic filler.\",\n \"This invention advantageously increases G′ of tire treads containing rubber fillers.\",\n \"Among other advantages, tire treads containing the modified rubbers of this invention also have improved dry traction.\",\n \"Another advantage of certain embodiments of this invention is the elimination of zinc oxide as an additive in tire rubber formulations.\"\n ],\n [\n \"FIELD OF THE INVENTION This invention relates to rubber compositions and tire components that have increased dynamic moduli, as well as tire components having improved stable modulus at higher temperatures.\",\n \"BACKGROUND OF THE INVENTION Hysteresis loss and storage modulus are properties of rubber that have a significant impact on tire performance.\",\n \"Hysteresis loss can be defined by the formula tan δ=G″/G′, where G″ is a measure of the dynamic loss modulus and G′ is the measure of storage modulus.\",\n \"Generally, as the temperature of a tire increase, G′ decreases.\",\n \"As a result, there may be a decrease in handling.\",\n \"Inorganic fillers, such as silica, impart improved wet traction (as demonstrated by a high tan δ at 0° C.) and improved snow traction (as demonstrated by a low tan δ at −20° C.) when used as filler within tire treads.\",\n \"Mixing silica into a tire stock, however, is difficult because silica particles agglomerate and therefore they are not easily dispersed.\",\n \"In response, processing and dispersing aids are used during compounding.\",\n \"Unfortunately, the use of these processing and dispersing aids enhances the decrease in modulus that is typically observed at high temperatures.\",\n \"Because inorganic fillers are technologically useful, there is a need to overcome the problems associated with the loss of G′ at elevated temperatures in tire treads containing inorganic fillers.\",\n \"SUMMARY OF THE INVENTION In general the present invention provides a tire component including a tire tread that comprises a vulcanized rubber, an inorganic filler, and a modified rubber, where the modified rubber i) contains pendent or functional groups that contain carboxylic acid or anhydride groups, ii) has pendently grafted thereto a polymerized metal salt of an unsaturated carboxylic acid, or iii) contains both i and ii.\",\n \"The present invention also includes a tire tread comprising a vulcanized rubber, from about 10 to about 100 parts by weight inorganic filler per 100 parts by weight rubber, and from about 0.2 to about 10 parts by weight modified rubber per 100 parts by weight rubber.\",\n \"The present invention also provides a tire component including a tire tread that comprises a vulcanized rubber, an inorganic filler, a modified polyolefin that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups and a modified rubber that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups.\",\n \"The present invention also includes a tire component including a tire tread that comprises a vulcanized rubber, an inorganic filler, a modified rubber that contains a polymerized metal salt of an unsaturated carboxylic acid, and wherein the tire component is devoid of zinc oxide.\",\n \"Additionally, the present invention also provides a tire prepared by a process comprising the steps of vulcanizing a rubber composition that has been prepared by compounding ingredients comprising i) a rubber, ii) an inorganic filler, iii) a modified rubber that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups or a polymerized metal salt of an unsaturated carboxylic acid and iv) optionally a modified polyolefin that contains pendent or terminal functional groups containing carboxylic acid or anhydride groups.\",\n \"The present invention also provides a tire component including a tire tread that comprises a vulcanized rubber, an inorganic filler, and a modified rubber, where the modified rubber is prepared by polymerizing i) an unsaturated carboxylic acid, ii) an unsaturated anhydride, iii) a metal salt of an unsaturated carboxylic acid or iv) combinations of i, ii or iii, to an elastomeric backbone via free-radical polymerization.\",\n \"The addition of poly(metal carboxylate) grafted rubbers to tire formulations including inorganic fillers has surprisingly been found to stabilize the dynamic modulus of the tire rubber at high temperatures.\",\n \"Also, the use of these poly(metal carboxylate) grafted rubbers has surprisingly lead to an increased tan δ at higher temperature, as well as decreased tire wear in tires filled with inorganic filler.\",\n \"This invention advantageously increases G′ of tire treads containing rubber fillers.\",\n \"Among other advantages, tire treads containing the modified rubbers of this invention also have improved dry traction.\",\n \"Another advantage of certain embodiments of this invention is the elimination of zinc oxide as an additive in tire rubber formulations.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS FIG.\",\n \"1 is a graphical plot of the temperature dependence of G′ of a vulcanizate prepared according to the present invention as compared to a control.\",\n \"FIG.\",\n \"2 is a graphical plot of the strain dependence of tan δ of a vulcanizate prepared according to the present invention as compared to a control.\",\n \"FIG.\",\n \"3 is a graphical plot of the strain dependence of G′ of a vulcanizate according to the present invention as compared to controls, where one control employs carbon black as a filler.\",\n \"DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS Tires and tire components are prepared from rubber compositions that include a) a rubber, b) a modified rubber, wherein said modified rubber i) contains pendent or functional groups that contain carboxylic acid or anhydride groups, ii) contains pendently grafted thereto a polymerized metal salt of an unsaturated carboxylic acid, or iii) both i and ii, c) a inorganic filler, and d) an optional modified polyolefin that contains pendent or terminal functional groups that contain carboxylic acid or anhydride groups.\",\n \"Other additives that are typically employed in rubber compositions, especially those used to fabricate tires, may also be included.\",\n \"Both synthetic and natural rubber may be employed within the rubber compositions.\",\n \"These rubbers, which may also be referred to as elastomers, include, without limitation, natural rubber, synthetic poly(isoprene), poly(styrene-co-butadiene), poly(butadiene), poly(styrene-co-butadiene-co-isoprene), poly(styrene-co-isoprene), and mixtures thereof.\",\n \"In the first embodiment, the modified rubber additive is an elastomeric polymer that contains terminal or pendent functional groups that contain carboxylic acid or anhydride groups.\",\n \"The terminal or pendent functional groups that contain the carboxylic acid or anhydride groups may be obtained by grafting functional groups to a polymeric chain or by preparing a copolymer by using at least one monomer that will yield the desired functional group The modified rubber additive can be obtained by polymerizing unsaturated carboxylic acids or unsaturated anhydrides from a graft point on an elastomeric polymer.\",\n \"For example, functional groups may be obtained by the free radical polymerization of methacrylic acid in the presence of the diene rubber.\",\n \"Examples of elastomers from which this grafting reaction may take place include poly(butadiene), poly(styreneco-butadiene), poly(isoprene), poly(styrene-co-butadiene-co-isoprene), poly(styrene-co-isoprene), and terpolymers of ethylene, an a-olefin, and diene monomers.\",\n \"Examples of saturated carboxylic acids that can undergo polymerization and graft to an elastomer include citraconic acid, cinnamic acid, methacrylic acid, itaconic acid, and acrylic acid.\",\n \"Examples of unsaturated anhydrides that can undergo polymerization and graft to an elastomer include maleic anhydride, citraconic anhydride and itaconic anhydride.\",\n \"Free radical polymerization is the preferred reaction for grafting these monomers to an elastomer.\",\n \"Preferably, this technique employs an initiator such as a thermo-decomposition initiator.\",\n \"Examples of these initiators include but are not limited to, di-sec-butyl peroxydicarbonate, t-amyl peroxy pivalate, 2,5-dimethyl-2,5-di-(2-ethylhexanoyl-peroxy) hexane, t-amylperoxy-2-ethylhexanoate, t-butyl-2-ethylhexanoate, 2,2-azo-bis-(2-methyl propionitrile), azo-bis-isobutyronitrile (AIBN) and the like.\",\n \"This grafting reaction preferably takes place in an inert solvent such as a hydrocarbon solvent.\",\n \"Where the functional group is grafted to an elastomeric polymer to form the modified rubber additive, the resulting grafted copolymer may contain from about 1 to about 50 percent by weight, preferably from about 2 to about 35 percent by weight, and even more preferably from about 3 to about 25 percent by weight of the grafted functional groups.\",\n \"Alternatively, the modified rubber can be obtained by copolymerizing unsaturated carboxylic acid monomers or unsaturated anhydride monomers with diene or other monomers that will provide a rubbery polymer.\",\n \"Examples of unsaturated carboxylic acids and unsaturated anhydrides are provided above.\",\n \"Examples of diene monomers that will yield a rubbery polymer include diene monomers such as 1,3-butadiene, and isoprene.\",\n \"Other monomer that may be copolymerized with these diene monomers include vinyl aromatic monomer such as styrene.\",\n \"This copolymerization technique is well-known, and it typically takes place in an emulsion polymerization with the use of a radical source such as a peroxide redox system.\",\n \"The number average molecular weight of the modified rubber additive of this embodiment can vary greatly, although it is preferred that the number average molecular weight (Mn) be from about 200,000 to about 500,000.The modified rubber additive of the first embodiment generally contains from about 0.02 to about 28 milliequivalents of functional groups (e.g., units deriving from methacrylic acid) per gram of modified polymer, preferably from about 0.1 to about 25, and more preferably from about 0.5 to about 20 milliquivalents of functional groups per gram of modified polymer.\",\n \"The second embodiment of the present invention is the poly(metal carboxylate) graft rubber, which is a macromolecular structure that comprises an unsaturated hydrocarbon rubber backbone having pendently grafted thereto a polymerized metal salt of an unsaturated carboxylic acid.\",\n \"Each macromolecular structure will contain at least one pendent graft, preferably at least 2 pendent grafts, and more preferably at least 5 pendent grafts per mole of rubber polymer chain.\",\n \"This grafted rubber is described in U.S. Pat.\",\n \"No.\",\n \"5,962,593, which is incorporated herein by reference.\",\n \"Any unsaturated hydrocarbon rubber may be utilized to form the polymeric backbone of the graft copolymer rubber.\",\n \"Preferred unsaturated hydrocarbon rubbers include homopolymers of conjugated dienes and copolymers of conjugated dienes and other monomers such as vinyl aromatics.\",\n \"Exemplary conjugated diene monomers include 1,3-butadiene, isoprene, and 2,3-dimethyl-1,3-butadiene.\",\n \"Exemplary vinyl aromatic monomers include styrene, α-methylstyrene, and p-methylstyrene.\",\n \"Other preferred hydrocarbon rubbers include terpolymers of ethylene, propylene, and dienes such as, but not limited to, 5-ethylidene-2-norbornene and 1,4-hexadiene.\",\n \"Some exemplary polymers include poly(butadiene), poly(styrene-co-butadiene), poly(ethylene-co-propylene-co-diene), poly(isoprene), poly(styrene-co-isoprene), and poly(butadiene-co-styrene-co-isoprene).\",\n \"Any unsaturated carboxylic acid may be used to form the metal salt.\",\n \"The unsaturated carboxylic acids include α,β-ethylenically unsaturated carboxylic acids having from 3 to 8 carbon atoms.\",\n \"Exemplary carboxylic acids include acrylic, methacrylic, cinnamic, and crotonic acids, with acrylic and methacrylic acids being preferred.\",\n \"Suitable metal ions that may be used to form the metal salts include sodium, potassium, magnesium, calcium, zinc, barium, aluminum, tin, zirconium, lithium and cadmium, with zinc and magnesium being preferred.\",\n \"The metal ion is preferably introduced in the form of a salt of the carboxylic acid.\",\n \"A preferred metal salt is zinc dimethacrylate.\",\n \"The poly(metal carboxylate) graft rubber can be prepared by (i) dissolving an unsaturated hydrocarbon rubber in a solvent, (ii) adding to the dissolved rubber a metal salt of an unsaturated carboxylic acid to form a rubber/carboxylic acid blend, (iii) adding to the rubber/carboxylic acid blend a free radical initiator, and (iv) heating the reaction mixture to a temperature of from about 40 to about 150° C. for about 0.1 to about 100 hours.\",\n \"Preferably, the reactor containing the rubber/carboxylic acid blend is sealed prior to adding the initiator.\",\n \"Once sealed, the initiator is preferably added in liquid form under pressure.\",\n \"The metal salt is preferably added as a fine powder with stirring and under a nitrogen atmosphere to form a dispersion of the metal salt in the polymer solution.\",\n \"Preferably, a surfactant is added along with the metal salt in order to provide a more stable dispersion of the metal salt in the polymer solution.\",\n \"Preferred surfactants include nonionic octylphenoxy polyethoxy ethanol surfactants, which available as Triton X-15, Triton X-45, and Triton X-100 (Rohm and Haas Co.; Philadelphia, Pa.).\",\n \"Preferably, any solvent in which both the rubber and the free radical initiator are soluble may be used.\",\n \"The preferred solvents are the hydrocarbon solvents in which the original polymers were prepared.\",\n \"Exemplary solvents are aromatic and aliphatic hydrocarbons including, but not limited to, hexane, heptane, pentane, octane, cyclohexane, cycloheptane, cyclopentane, methyl cyclohexane, benzene, and toluene.\",\n \"Hexane and toluene are preferred.\",\n \"Suitable free radical initiators include, but are not limited to, di-sec-butyl peroxydicarbonate, t-amyl peroxy pivalate, 2,5-dimethyl-2,5-di-(2-ethylhexanoyl-peroxy) hexane, t-amylperoxy-2-ethylhexanoate, t-butyl-2-ethylhexanoate, 2,2-azo-bis-(2-methyl propionitrile), azo-bis-isobutyronitrile (AIBN) and the like.\",\n \"The optional modified polyolefin additive includes an α-olefin polymer that contains terminal or pendent moieties containing acid or anhydride groups.\",\n \"The terminal or pendent moieties typically derive from unsaturated carboxylic acids or unsaturated anhydrides.\",\n \"Examples of unsaturated carboxylic acids include citraconic acid, cinnamic acid, methacrylic acid, and itaconic acid.\",\n \"Examples of unsaturated anhydrides include maleic anhydride, citraconic anhydride, and itaconic anhydride.\",\n \"The preferred terminal or pendent moieties are succinic anhydride groups, or the corresponding acid from a ring opening structure, that derives from maleic anhydride.\",\n \"The α-olefin polymer includes an α-olefin homopolymer, a copolymer of two or more α-olefins, or a copolymer of an α-olefin with a compatible monomer.\",\n \"The α-olefins can include from about 2 to about 8 carbon atoms, and more preferably from 3 to about 5 carbon atoms.\",\n \"Exemplary α-olefin monomers include ethylene, propylene, butene-1, and pentene-1.Exemplary monomers that can be copolymerized with α-olefins include vinyl aromatic monomers and diene monomers.\",\n \"An exemplary copolymer is poly(propylene-co-ethylene) that contains polyethylene crystals.\",\n \"The modified polyolefins should contain from about 0.01 to about 3 pbw of the functional moiety based upon the weight of the entire polymer.\",\n \"More preferably, the modified polyolefin should contain from about 0.1 to about 2 pbw of the functional moiety, an even more preferably from about 0.15 to about 1.0 pbw of the functional moiety.\",\n \"The number average molecular weight of the modified polyolefins can vary greatly, although it is preferred that the number average molecular weight (Mn) be from about 20,000 to about 500,000, more preferably from about 100,000 to about 400,000, and even more preferably from about 150,000 to about 400,000, as determined by using standard GPC analysis with polystyrene as a standard.\",\n \"Generally, the molecular weight distribution (Mw/Mn) should be less than about 4.5, preferably less than about 4.0, and even more preferably less than about 3.8.The modified polyolefin additives are typically prepared by grafting unsaturated carboxylic acids or unsaturated anhydrides to a polyolefin polymer.\",\n \"The techniques employed to attach the terminal or pendent moieties that contain carboxylic acid or anhydride groups to a polyolefin polymer are well known in the art.\",\n \"For example, grafting maleic anhydride to a polyolefin is disclosed in U. S. Pat.\",\n \"No.\",\n \"6,046,279, which is incorporated herein by reference.\",\n \"The α-olefin polymers can be synthesized by using a number of polymerization techniques such as, but not limited to, the “Phillips catalyzed reactions” conventional Ziegler-Natta type polymerizations, and metallocene catalysis including, but not limited to, metallocene-aluminoxane and metallocene-ionic activator catalysis.\",\n \"Exemplary α-olefin polymers include polyethylene, polypropylene, poly(ethylene-co-propylene), poly(propylene-co-butene-1), and poly(butene-1-co-ethylene).\",\n \"These α-olefin polymers can be either amorphous, semi-crystalline, or crystalline polymers.\",\n \"The preferred polyolefins include crystalline or stereoregular polypropylene.\",\n \"Most polypropylene homopolymers that are commercially produced have an isotactic microstructure.\",\n \"The poly(propylene-co-ethylene) copolymers can be random or block copolymers.\",\n \"Preferably, these copolymers will contain some polyethylene crystals, although they should include a major amount of propylene units and only a minor amount of ethylene units.\",\n \"Preferably, these copolymers should contain less than about 40 percent by weight (pbw) ethylene units, more preferably from about 1 to about 30 pbw ethylene units, and more preferably from about 1.5 to about 25 pbw ethylene units.\",\n \"Modified polyolefins are commercially available.\",\n \"For example, maleic anhydride functionalized polypropylene is available under the tradename EXXELOR™ P01015 & 1020 (Exxon Mobil Chemical Company; Houston, Tex.\",\n \"), under the tradename PP-C™, CA1000, or 18707 (Elf Atochem; Philadelphia, Pa.), or under the tradename Polybond™ 3001, 3002, or 3150 (Uniroyal Chemical Company; Middlebury, Conn.).\",\n \"The rubber compositions preferably include an inorganic filler as well as optional organic fillers.\",\n \"The inorganic fillers may include silica, aluminum hydroxide, magnesium hydroxide, and clays (hydrated aluminum silicates).\",\n \"The optional organic fillers includes carbon black.\",\n \"Useful silica (silicon dioxide) includes wet-process, hydrated silica produced by a chemical reaction in water, and precipitated as ultrafine spherical particles.\",\n \"These particles strongly associate into aggregates that in turn combine less strongly into agglomerates.\",\n \"The surface area, as measured by the BET method, gives the best measure of the reinforcing character of different silicas.\",\n \"Useful silicas preferably have a surface area of about 32 to about 400 m2/g, preferably about 100 to about 250 m2/g, and more preferably about 150 to about 220 m2/g.\",\n \"The pH of the silica filler is generally about 5.5 to about 7 and preferably about 5.5 to about 6.8.Commercially available silicas include Hi-Sil™ 215, Hi-Sil™ 233, and Hi-Sil™ 190 (PPG Industries; Pittsburgh, Pa.).\",\n \"Useful commercial grades of different silicas are also available from other sources including Rhone Poulenc.\",\n \"Useful aluminum hydroxide fillers include any aluminum hydroxide filler including those that have conventionally been employed to reinforce rubber compositions and tire components.\",\n \"Aluminum hydroxide filler preferably has an average diameter of about 20 nanometers (nm) to about 2,000 nm, more preferably from about 25 nm to about 1,000 nm or smaller, and even more preferably from about 30 nm to about 50 nm.\",\n \"Useful aluminum hydroxide preferably has a BET specific surface area of from about 0.5 to about 500 m2/g, more preferably from about 1 to about 400 m2/g, and even more preferably from about 2 to about 300 m2/g.\",\n \"Aluminum hydroxide filler may be treated with various surface treating agents.\",\n \"Non-limiting examples of surface treating agents include organic treating agents such as fatty acids and alcohols, and inorganic dispersants such as coupling agents and fatty acid metal salts.\",\n \"Useful aluminum hydroxide filler is commercially available from a number of sources including that available under the tradenames Hygilite™ (Showa Dendo K.K.\",\n \"; Japan), Baikalox™(Baikowski Chimie; France), and RF22™ (Sumitono Chemical Co.; Japan).\",\n \"Useful carbon black may include any commonly available carbon black, but those having a surface area (EMSA) of at least 20 m2/g, and more preferably at least 35 m2/g up to 200 m2/g or higher, are preferred.\",\n \"Surface area values used in this application are those determined by ASTM test D-1765 by using the cetyltrimethyl-amnmonium bromide (CTAB) technique.\",\n \"A coupling agent may optionally be added when an inorganic filler such as silica is used.\",\n \"Coupling agents include those compounds that are believed to react with both the rubber and the inorganic filler.\",\n \"One coupling agent conventionally used is bis-[3(triethoxysilyl) propyl]-tetrasulfide, which is commercially available under the tradename S169 (Degussa, Inc.; New York, N.Y.).\",\n \"Other coupling agents include bis-[3(tdethoxysilyl) propyl]-disulfide, which is commercially available under the tradename Silquest™ (Witco; Greenwich, Conn.), mercapto propyl alkoxy silane, which is commercially available under the tradename Ciptane™ (Dow Corning; Midland, Mich.), and dithio diproprionic acid or carboxylic acid disulfides, which are commercially available from Aldrich Chemical Company.\",\n \"In general, these coupling agents should be used in an amount from about 0.1 to 20% by weight based upon the weight of the inorganic filler.\",\n \"Shielding and dispersing agents, which prevent or alleviate the agglomeration of inorganic filler particles such as silica, may also be used.\",\n \"Typically, these agents react or interact with the filler.\",\n \"Exemplary dispersing or shielding agents include silanes, amines, diols, polyethers, amides, and sugar fatty acid esters.\",\n \"U.S. Pat.\",\n \"Nos.\",\n \"5,719,207, 5,780,538, 5,717,022, and EP 0890606 are incorporated herein by reference in this regard.\",\n \"Specific examples of these agents include sugar, fatty acids such as sorbitan fatty acids which are available from BASF (Mount Olive, N.J.), and octyl triethoxy silane, which is available from Dow Corning (Midland, Mich.).\",\n \"Generally, these shielding or dispersing agents may be used in an amount from about 0.1 to about 20% by weight based on the weight of the inorganic filler.\",\n \"In preferred embodiments, the coupling agents, shielding agents, and dispersing agents may be used in combination.\",\n \"The rubber compositions of this invention can be cured in a conventional manner with known vulcanizing agents.\",\n \"For example, sulfur or peroxide-based curing systems may be employed.\",\n \"Also, see Kirk-Othmer, ENCYCLOPEDIA OF CHEMICAL TECHNOLOGY, 3rd Edition, Wiley Interscience, N.Y. 1982, Vol.\",\n \"20, pp.\",\n \"365-468, particularly VULCANIZATION AGENTS AND AUXILIARY MATERIALS pp.\",\n \"390-402, or Vulcanization by A. Y. Coran, ENCYCLOPEDIA OF POLYMER SCIENCE AND ENGINEERING, 2nd Edition, John Wiley & Sons, Inc., 1989, which are incorporated herein by reference.\",\n \"Vulcanizing agents may be used alone or in combination.\",\n \"This invention does not appreciably affect cure times.\",\n \"Typically, vulcanization is effected by heating the vulcanizable composition; e.g., it is heated to about 170° C.. Cured or crosslinked polymers may be referred to as vulcanizates, which are generally three-dimensional polymeric networks that are thermoset.\",\n \"Depending on the degree of unsaturation of the modified rubber, it may become part of the crosslinked network comprising the vulcanizate.\",\n \"Alternatively, if the modified rubber is highly saturated, it will be dispersed within the crosslinked polymeric network.\",\n \"The other ingredients, such as the modified polyolefin and fillers, are generally dispersed throughout this thermoset network.\",\n \"The rubber compositions and tire components may also include other compounding additives such as accelerators, oils, waxes, scorch inhibiting agents, and processing aids.\",\n \"Compositions containing synthetic rubbers typically include antidegradants, processing oils, zinc oxide, optional tackifying resins, optional reinforcing resins, optional fatty acids, optional peptizers, and optional scorch inhibiting agents.\",\n \"The use of the poly(metal carboxylate) graft rubber according to this invention can, advantageously, eliminate the need for ingredients such as zinc oxide and stearic acid.\",\n \"The use of the terminal or pendent functional groups that contain the carboxylic acid or anhydride groups with the modified polyolefin can also eliminate the need for ingredients such as zinc oxide.\",\n \"Accordingly, the rubber compositions of certain embodiments of this invention may be essentially devoid of zinc oxide and stearic acid, where essentially devoid refers to the absence of any amount that is conventionally required to impact compounding and curing.\",\n \"The rubber compositions and tire components of the present invention will generally contain from about 0.1 to about 25 parts by weight of the modified rubber additive per 100 parts by weight rubber (phr).\",\n \"Preferably, the rubber compositions and tire components will contain from about 0.2 to about 20 parts by weight phr, even more preferably from about 0.5 to about 15 parts by weight phr, and still more preferably from about 1 to about 10 parts by weight phr of the modified rubber additive.\",\n \"The rubber compositions and tire components of the present invention will optionally contain from about 0.1 to about 25 parts by weight of the modified polyolefin additive per 100 parts by weight rubber (phr).\",\n \"Preferably, the rubber compositions and tire components will contain from about 0.2 to about 20 parts by weight phr, even more preferably from about 0.5 to about 15 parts by weight phr, and still more preferably from about 1 to about 10 parts by weight phr of the modified polyolefin additive.\",\n \"Fillers, such as carbon black, silica or aluminum hydroxide, are typically employed in an amount from about 1 to about 100 parts by weight phr, and preferably from about 20 to about 90 parts by weight phr, and more preferably from about 40 to about 80 parts by weight phr.\",\n \"Silica may optionally be used in an amount from about 10 to about 100 parts by weight phr, preferably from about 15 to about 90 parts by weight phr, and more preferably from about 20 to about 80 parts by weight phr.\",\n \"Carbon black may optionally be used in an amount from about 0.5 to about 80 parts by weight phr, preferably from about 1 to about 40 parts by weight phr, and more preferably from about 2 to about 30 parts by weight phr.\",\n \"Optionally, aluminum hydroxide filler should be employed in an amount from about 1 to about 25 parts by weight phr, preferably from about 2 to about 20 parts by weight phr, and more preferably from about 5 to about 15 parts by weight phr.\",\n \"In a preferred embodiment, aluminum hydroxide, silica, and carbon black are used in combination.\",\n \"In this embodiment, the silica:carbon black ratio is from about 1:10 to about 99:1, more preferably from about 1:5 to about 5:1, and even more preferably from about 1:3 to about 3:1.And, the silica:aluminum hydroxide ratio is from about 3:1 to about 30:1, more preferably from about 5:1 to about 20:1, and even more preferably from about 6:1 to about 15:1.Those skilled in the art will be able to choose a useful amount of the other ingredients that may be employed in practicing the present invention.\",\n \"For example, it is generally known in the art of making tire components, such as treads, that sulfur is typically employed in an amount from about 0.5 to about 10 parts by weight phr, and preferably from about 1 to about 6 parts by weight phr.\",\n \"Oils are typically employed in an amount from about 1 to about 60 parts by weight phr, and preferably in an amount from about 1 to about 50 parts by weight phr.\",\n \"Zinc oxide is typically employed in an amount from about 0.5 to about 8, and preferably from about 1 to about 5 parts by weight phr, however, compositions which contain the poly(metal carboxylate) graft rubber according to this invention, or modified rubber having terminal or pendent functional groups that contain the carboxylic acid or anhydride groups and the modified polyolefin of the present invention may eliminate the need for ingredients such as zinc oxide and stearic acid.\",\n \"Tire formulations are compounded by using mixing equipment and procedures conventionally employed in the art.\",\n \"Preferably, an initial masterbatch is prepared that includes the elastomer component and the reinforcing fillers, as well as other optional additives such as processing oil and antioxidants.\",\n \"The modified rubbers are preferably added during preparation of the initial masterbatch.\",\n \"Once this initial masterbatch is prepared, the vulcanizing agents are blended into the composition.\",\n \"Rubber compounding techniques and the additives employed therein are generally known as disclosed in The Compounding and Vulcanization of Rubber, by Stevens in RUBBER TECHNOLOGY SECOND EDITION (1973 Van Nostrand Reihold Company).\",\n \"The mixing conditions and procedures applicable to silica-filled tire formulations are also well known as described in U.S. Pat.\",\n \"Nos.\",\n \"5,227,425, 5,719,207, 5,717,022, as well as EP 0890606, all of which are incorporated herein by reference.\",\n \"The composition can then be processed into tire components according to ordinary tire manufacturing techniques including standard rubber curing techniques.\",\n \"Tire components of this invention preferably include tire treads.\",\n \"The compositions, however, can also be used to form other elastomeric tire components such as subtreads, black sidewalls, body ply skims, bead fillers and the like.\",\n \"Pneumatic tires can be made according to U.S. Pat.\",\n \"Nos.\",\n \"5,866,171; 5,876,527; 5,931,211; and 5,971,046, which are incorporated herein by reference.\",\n \"Tire components of this invention preferably include tire treads.\",\n \"The composition can also be used to form other elastomeric tire components such as subtreads, sidewalls, body ply skims, bead fillers and the like.\",\n \"The use of polyolefin additives within tires treads that contain the modified rubbers of the present invention leads to both reduced hysteresis loss while improving handling performance without the need for other additives such as particulate nylons.\",\n \"In order to demonstrate the practice of the present invention, the following examples have been prepared and tested as described in the General Experimentation Section disclosed hereinbelow.\",\n \"The examples should not, however, be viewed as limiting the scope of the invention.\",\n \"The claims will serve to define the invention.\",\n \"EXAMPLES Example 1 Stocks 1-2 Two tire stocks were prepared from the formulations in Table I by using the compounding protocol set forth in Table II within a 310 gram Brabender mixer set at 60 r.p.m.\",\n \"TABLE I Stock 1 Stock 2 Polybutadiene 100 90 Poly(butadiene-g-methacrylic acid) 0 10 Precipitated silica 65.00 65.00 Aromatic Process oil 16.25 16.25 Wax 1.5 1.5 Antioxidant [N-(1,3 dimethylbutyl)-N′-phenyl-p- 1.70 1.70 phenylene-diamine] Si69 6.5 6.5 Sulfur 2.85 2.85 Steric acid 2.00 2.00 Accelerator [N-Cyclohexyl-2-benzothiazyl- 1.50 1.50 sulfenamide] Diphenyl Guanidine 0.50 0.50 Zinc oxide 2.5 2.5 The polybutadiene was obtained under the tradename HD55™ (Firestone Polymers; Akron, Ohio).\",\n \"The poly(butadiene-g-methacrylic acid) was prepared by grafting unsaturated methacrylic acid to polybutadiene by using azobisisobutyronitrile (AIBN) as an initiator.\",\n \"This grafting took place in a hydrocarbon solvent at 60-80° C. with 10 parts by weight methacrylic acid per 100 parts by weight polybutadiene.\",\n \"The polybutadiene had a Mooney Viscosity (ML1+4 at 100° C.) of 52 and a cis microstructure of about 33 weight percent.\",\n \"The poly(butadiene-g-methacrylic acid) had a Mooney Viscosity (ML1+4 at 100° C.) of 56, a cis microstructure of about 33 weight percent, and contained about 10 weight percent units deriving from the methacrylic acid.\",\n \"TABLE II Master Batch Stage Initial Temperature 100° C. 0 sec charging polymers 30 sec charging silica and all pigments 5 min drop Drop Temperature 155° C. Remill 1 Batch Stage Initial Temperature 70° C. 0 sec charging remilled stock 30 sec charging Si69 Drop Temperature 155° C. Final Batch Stage Initial Temperature 90° C. 0 sec charging remilled stock 30 sec charging curing agent and accelerators Drop Temperature 105° C. The final stocks were sheeted and then subsequently molded at 171° C. for 15 minutes.\",\n \"The Mooney Viscosity, δ(ΔG′), and curing characteristics of the uncured stocks were evaluated.\",\n \"Cured samples of each stock were also evaluated for tensile mechanical properties at 25° C. Further, the dynamic viscoelastic properties of cured samples of each stock were also evaluated.\",\n \"The results of this analysis are set forth in Table II.\",\n \"TABLE III Stock No.\",\n \"1 2 Uncured Analysis Mooney Viscosity (ML1+4 @ 100° C.) 80.8 81.7 T5 Scorch @ 130° C. (sec) 951 1,037 tS2 @ 171° C. (min) 1.07 1.18 T90 @ 171° C. (min) 6.82 6.77 δ(ΔG′) (kPa) 2,062 1,106 Tensile Properties Modulus @ 50% (MPa)) 1.63 1.79 Modulus @ 300% (MPa) 9.86 10.74 Tensile Strength @ Break (MPa) 10.90 12.64 Elongation @ Break (%) 314 333 Toughness (MPa) 14.82 17.97 Viscoelastic Properties Tanδ @ 0° C. (temperature sweep) 0.1733 0.1612 Tanδ @ 50° C. (temperature sweep) 0.1245 0.1131 ΔG′ (G′@.25%-G′@14.75%) 0.253 0.2414 @ 65° C. (strain sweep) (MPa) Tanδ @ 7% strain and 65° C. 0.1055 0.1026 (strain sweep) British Portable Skid Tester 42 42 Abrasion Resistance (%) 100 111 The Mooney viscosity measurement was conducted at 130° C. by using a larger rotor, and was recorded as the torque when the rotor has rotated for 4 minutes.\",\n \"Each sample was preheated at 130° C. for one minute before the rotor started.\",\n \"T5 is the time required to increase 5 Mooney units during the Mooney-scorch measurement.\",\n \"This index is used to predict how fast the compound viscosity will rise during processing.\",\n \"tS2 and T90 are the times required for torque rises to 2% and 90% of the total torque increase during curing characterization experiments at 171° C. These measurements are typically used to predict how fast the viscosity build-up (tS2) and the curing rate during the curing process.\",\n \"The G′ of the uncured stocks was obtained from a strain sweep measurement by using an RPA2000 Rubber Process Analyzer (Alpha Technologies).\",\n \"These strain sweep experiments were conducted at 50° C. at a frequency of 0.1 Hz with strain sweeping from 0.25% to 1,000%.\",\n \"The difference in G′ at strains of 0.25% and 1,000% were recorded as the ΔG′.\",\n \"ΔG′ was also determined after the stocks were annealed at 171° C. for 15 minutes.\",\n \"δ(ΔG′), therefore, is the difference between ΔG′ before annealing and ΔG′ after annealing (ΔG′ after heat treatment−ΔG′ before heat treatment).\",\n \"The tensile mechanical properties were measured by using the standard procedure described in ASTM-D 412 at 25° C. The tensile test specimens were round rings with a dimension of 1.27 mm in width and 1.905 mm in thickness.\",\n \"A specific gauge length of 25.4 mm was used for the tensile test.\",\n \"Temperature sweep experiments were conducted with a frequency of 31.4 rad/sec by using 0.5% stain for temperatures ranging from −100° C. to −10° C., and 2% strain for temperatures ranging from −10° C. to 100° C. Payne effect (ΔG′) and tanδ at 7% strain were obtained from the strain sweep experiments.\",\n \"A frequency of 3.14 rad/sec was used for strain sweep experiments conducted at 65° C. with strain sweeps from 0.25% to 14.75%.\",\n \"The wet traction of the rubber was evaluated by using the British Portable Skid Tester.\",\n \"The details of this test are described in ASTM E-303, Vol.\",\n \"04.03.According to this test, a specimen is attached to the base of a pendulum arm, and the specimen contacts the opposing surface during a swing of the pendulum.\",\n \"The weighted pendulum head is free to move vertically on the pendulum arm so that the swing amplitude determines the friction of the rubber against the pavement surface.\",\n \"The lower the amplitude that the pendulum swings up after contacting the surface (recorded as a higher value on the scale of the tester), the higher friction of the rubber against the surface.\",\n \"The wear resistance of the cured samples were evaluated by weighing the amount of wear according to the Lambourne test.\",\n \"The wearing index was obtained from the ratio of the weight loss of the control to that of the testing sample.\",\n \"Samples with higher wear indices have better wear resistance properties.\",\n \"Samples used for this Lambourne test are circular donuts with the following approximate dimensions: 22.86 mm and 48.26 mm for the inside and outside diameters, and 4.95 mm in thickness.\",\n \"The test specimens were place on an axil and run at a slip ratio of 25% against a driven abrasive surface.\",\n \"Stocks 3-6 Four tire stocks were prepared from the formulations in Table IV by using the compounding protocol set forth in Table V within a 310 gram Brabender mixer set at 60 r.p.m.\",\n \"TABLE IV Stock 3 Stock 4 Stock 5 Stock 6 Polybutadiene 90 90 100 90 poly(butadiene-g-methacrylic 10 10 0 10 acid) Polypropylene 0 10 0 00 poly((propylene-co-ethylene)- 0 0 10 10 g-maleic anhydride) Precipitated silica 65 65 65 65 Aromatic Process oil 16.25 16.25 16.25 16.25 Wax 1.5 1.5 1.5 1.5 Antioxidant [N- 0.95 0.95 0.95 0.95 (1,3 dimethylbutyl)- N′-phenyl-p-phenylene- diamine] Si69 6.50 6.50 6.50 6.50 Sulfur 2.85 2.85 2.85 2.85 Steric acid 1.50 1.50 1.50 1.50 Accelerator [N-Cyclohexyl-2- 1.50 1.50 1.50 1.50 benzothiazyl-sulfenamide] Diphenyl Guanidine 0.5 0.5 0.5 0.5 Zinc oxide 2.5 2.5 2.5 2.5 The polybutadiene and poly(butadiene-g-methacrylic acid) were the same as those from Table I.\",\n \"This grafting took place under substantially the same conditions as used for Samples 1 and 2.The polypropylene was obtained under the tradename PP340™ (Aldrich) and was characterized by having a weight average molecular weight (Mw) of about 340,000, a melt temperature (Tm) of about 161° C., and a crystallinity of greater than about 75%.\",\n \"The poly((propylene-co-ethylene)-g-maleic acid) was obtained under the tradename Orevac™ 18707 (Elf Atochem) and was characterized by having a Mw of about 360,000, a Tm of about 120° C. and about 150° C., a crystallinity that was greater than about 75%, an ethylene content of about 21 weight percent, and contained about 0.48 weight percent of moieties deriving from maleic anhydride.\",\n \"TABLE V Master Batch Stage Initial Temperature 100° C. 0 sec charging polymers and polyolefin (if added) 30 sec charging silica and all pigments 5 min drop Drop Temperature 175° C. Remill 1 Batch Stage Initial Temperature 70° C. 0 sec charging remilled stock charging rest of the silica 30 sec charging Si69 Drop Temperature 155° C. Final Batch Stage Initial Temperature 90° C. 0 sec charging remilled stock 30 sec charging curing agent and accelerators Drop Temperature 105° C. The final stocks were sheeted and then subsequently molded at 171° C. for 15 minutes.\",\n \"The Mooney Viscosity, δ(ΔG′), and curing characteristics of the uncured stocks were evaluated.\",\n \"Cured samples of each stock were also evaluated for tensile mechanical properties at 25° C. and after heat aging at 100° C. for 24 hours.\",\n \"Further, the dynamic viscoelastic properties of cured samples of each stock were also evaluated.\",\n \"The results of this analysis are set forth in Table VI.\",\n \"TABLE VI Stock No.\",\n \"3 4 5 6 Uncured Analysis Mooney Viscosity (ML1+4 @ 100° C.) 81.7 110.7 98.4 101.1 T5 Scorch @ 130° C. (min) 1,037 874 926 930 tS2 @ 171° C. (min) 1.18 1.37 1.27 1.23 T90 @ 171° C. (min) 6.77 6.47 6.79 6.90 Δ(ΔG′) (kPa) 1,106 3,224 1,622 987 Tensile Properties (ring-shaped samples) Modulus @ 50% (MPa) 1.79 2.56 2.10 2.12 Modulus @ 300% (MPa) 10.74 13.56 11.09 11.16 Tensile Strength @ Break (MPa) 12.64 16.55 14.24 16.02 Elongation @ Break (%) 333 353 361 389 Toughness (MPa) 17.97 27.54 23.74 27.98 Tensile Properties (dumbbell-shaped samples) Modulus @ 50% (MPa) 1.79 3.08 2.37 2.33 Modulus @ 300% (MPa) 7.52 11.75 9.43 9.25 Tensile Strength @ Break (MPa) 13.73 16.35 14.15 16.28 Elongation @ Break (%) 511 432 448 518 Toughness (MPa) 33.90 38.00 33.00 42.46 Viscoelastic Properties Tanδ @ 0° C. (temperature sweep) 0.1612 0.1754 0.1744 0.1689 Tanδ @ 50° C. (temperature sweep) 0.1131 0.1255 0.1206 0.1217 ΔG′ (G′@.25%-G′@14.75%) @ 65° C. (strain sweep) (MPa) 0.79 1.37 0.98 1.085 Tanδ @ 7% strain and 65° C. (strain sweep) 0.2414 0.476 0.3813 0.3762 British Portable Skid Tester 0.1026 0.1115 0.1106 0.1120 Abrasion Resistance (%) 100 100 109 110.4 TABLE VII Stock No.\",\n \"3 4 5 6 Tensile Properties @ 25° C. (after thermal aging) Modulus @ 50% (MPa) 2.37 3.61 2.85 2.79 Tensile Strength @ Break 14.15 16.86 15.79 17.24 (MPa) Elongation @ Break (%) 249 241 258 286 Toughness (MPa) 15.34 19.71 18.24 22.21 Tear Properties @ 100° C. Strength (MPa) 1.84 1.70 1.98 2.22 Elongation @ Break (%) 284 192 290 320 Stocks 7-8 Two tire stocks were prepared from the formulations of Table VIII (expressed in parts by weight) by using the compounding protocol set forth in Table II within a 310 gram Brabender mixer set at 60 r.p.m.\",\n \"TABLE VIII Stock 7 Stock 8 Natural Rubber 20.0 20.0 Solution SBR 50.0 50.0 Poly(metal carboxylate) grafted rubber 0.0 5.0 Oil extended solution (poly(styrene-co- 27.5 27.5 butadiene) Poly(butadiene) 10.0 10.0 Carbon Black 33.3 33.3 Precipitated silica 33.0 33.0 Wax 1.0 1.0 Antioxidant [N-(1,3 dimethylbutyl)-N′-phenyl- 0.95 0.95 p-phenylene-diamine Sulfur 2.7 2.7 Accelerator [N-Cyclohexyl-2- 0.5 0.5 benzothiazolsulfenamine] Zinc oxide 1.5 1.5 Diphenyl Guanidine 0.5 0.5 Dioctyl phthalate 6.0 6.0 Aromatic processing oil 5.0 5.0 N-octyl triethoxy silane 1.8 1.8 Sorbitan Monooleate 5.0 5.0 Si69 0.28 0.28 The solution SBR was obtained by polymerizing 1,3-butadiene monomer and styrene monomer in hexane solvent with an alkyllithium initiator.\",\n \"The living polymers were then coupled with tin tetrachloride.\",\n \"The resulting polymers had a Mooney Viscosity (ML1+4 at 100° C.) of about 50, a vinyl content of about 58%, and a styrene content of about 20%.\",\n \"The oil extended solution poly(styrene-co-butadiene) was similarly prepared and extended in oil.\",\n \"The poly(metal carboxylate) grafted rubber was prepared by grafting zinc methacrylate to a poly(butadiene) polymer that was obtained under the tradename DIENE 55™ (Firestone Synthetic Polymers; Akron, Ohio) by using an AIBN initiator.\",\n \"The resulting grafted rubber had about 33% by weight poly(zinc methacrylate) graphs.\",\n \"The final compounds were sheeted and cured.\",\n \"Test specimens for dynamic storage moduli experiments were formed.\",\n \"The dynamic storage moduli were measured by temperature sweep experiments.\",\n \"These measurements were conducted with a frequency of 31.4 rad/sec by using 0.5 percent strain for temperatures ranging from −100° C. to −10° C., and 2 percent strain for the temperature ranging from −10° C. to 100° C. A frequency of 3.14 rad/sec was used for strain sweep, which was conducted at 65° C. with strain sweeping from 0.25 percent to 14.75 percent.\",\n \"The temperature dependence of the G′ of stocks 1 and 2 are shown in FIG.\",\n \"1.The vulcanizates of stocks 7 and 8 were also subjected to strain sweep experiments.\",\n \"These experiments were conducted at a frequency of 3.14 rad/sec was used for strain sweep, which was conducted at 65° C. with strain sweeping from 0.25 percent to 14.75 percent.\",\n \"From this data, tan δ was calculated and plotted over the strain sweep as shown in FIG.\",\n \"2.The vulcanizates were then tested for tensile properties according to ASTM D 412 at 25° C. The tensile test specimens were round rings with a diameter of 0.127 cm and a thickness of 0.191 cm.\",\n \"A specific gauge length of 2.54 cm was used.\",\n \"Table III provides the results of the tests.\",\n \"The vulcanizates of stocks 7 and 8 were also evaluated for wear resistance by the Lambourne test.\",\n \"Samples of this test were circular donuts with the following approximate dimensions: 22.86.mm inner diameter and 48.26 mm outer diameter, and 4.95 mm thick.\",\n \"Test specimens were placed on an axle and run at a slip ratio of 25% against a driven abrasive surface.\",\n \"The wear indices were obtained from the ratio of the weight loss of the control to that of the tested sample.\",\n \"Samples with higher wear indices have better wear resistance properties.\",\n \"Table IX provides the results of these tests.\",\n \"TABLE IX Stock 7 Stock 8 50% Modulus (MPa) 1.39 1.30 300% Modulus (MPa) 10.36 9.302 Tensile at Break (MPa) 16.27 18.14 Elongation at break (%) 419 485 Toughness (MPa) 29.60 39.96 Lambourne Weight loss (g) 0.1108 0.0808 Lambourne Wear Index 100 127 Stocks 9-11 Three tire stocks were prepared from the formulations in Table X by using the compounding protocol set forth in Table II within a 310 g Brabender mixer set at 60 rpm.\",\n \"TABLE X Stock 9 Stock 10 Stock 11 Natural Rubber 20.0 20.0 20.0 Solution SBR with modified chain ends 50.0 50.0 50.0 Poly(metal carboxylate) grafted rubber 0.0 5.0 5.0 Oil extended solution (poly(styrene-co- 27.5 27.5 27.5 butadiene) Poly(butadiene) 10.0 10.0 10.0 Carbon Black (SAF) 33.3 33.3 66.4 Precipitated silica 33.0 33.0 0 Wax 1.0 1.0 1.0 Antioxidant [N-(1,3 dimethylbutyl)-N′- 0.95 0.95 .95 phenyl-p-phenylene-diamine Sulfur 2.7 2.7 2.7 Accelerator [N-Cyclohexyl-2- 0.5 0.5 0.5 benzothiazolsulfenamine] Zinc oxide 1.5 1.5 1.5 Diphenyl Guanidine 0.5 0.5 0.5 Dioctyl phthalate 6.0 6.0 6.0 Aromatic processing oil 5.0 5.0 5.0 Octyl triethyl silane 1.8 1.8 1.8 Sorbitan Monooleate 5.0 5.0 5.0 Si69 0.28 0.28 0.28 The ingredients set forth in Table X were essentially the same as the ingredients set forth in Table VIII.\",\n \"TABLE XI Master Batch Stage 1 Initial Temperature 100° C. 0 sec charging polymers 30 sec charging carbon black, silica, shielding agent or Silanes (if added) and all pigments.\",\n \"5 min.\",\n \"drop Drop temperature 175-180° C. Master Batch Stage 2 Initial Temperature 70° C. 0 sec charging remilled stock 30 sec charging Si69 (if added) and remaining silica.\",\n \"Drop Temperature 155° C. Test specimens for dynamic storage moduli experiments were prepared in a similar fashion to those prepared for Stocks 7 and 8, and they were likewise analyzed in a similar fashion.\",\n \"The temperature dependence of G′ for Stocks 9-11 is shown in FIG.\",\n \"3.Various modifications and alterations that do not depart from the scope and spirit of this invention will become apparent to those skilled in the art.\",\n \"This invention is not to be duly limited to the illustrative embodiments set forth herein\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473912"}}},{"rowIdx":764,"cells":{"text":{"kind":"list like","value":[["Method and apparatus for fast w-cdma acquisition","Apparatus for classifying a ray is disclosed, wherein the ray is received at a code division multiple access receiver.","The apparatus comprises: means for receiving a neighbouring cell list comprising at least information concerning scrambling codes used by at least one neighbouring cell; means for selecting a scrambling code used by a given neighbouring cell; means for mixing a received signal with the selected scrambling code to provide a mixed signal; and means for examining the mixed signal to determine if the ray originates from the given neighbouring cell."],["1.Apparatus for classifying a ray received at a code division multiple access receiver, the apparatus comprising; means for receiving a neighbouring cell list comprising at least information concerning scrambling codes used by at least one neighbouring cell; means for determining whether a received unclassified ray has a timing characteristic falling within a predetermined range of a timing characteristic of a ray which has been classified to a selected neighbouring cell; means for mixing the signal corresponding to the received unclassified ray with a scrambling code used by said selected neighbouring cell thereby to provide a mixed signal, the scrambling code being received from the neighbouring cell list; and means for examining the mixed signal to determine if the ray originates from said selected neighbouring cell, the mixing and examining means being arranged to operate only if it is determined that the timing characteristic of the unclassified ray falls within the predetermined range.","2.Apparatus according to claim 1, further comprising means arranged to select a different cell from the neighbouring cell list in the event that it is determined that the timing characteristic of the unclassified ray falls outside the predetermined range.","3.Apparatus according to claim 1 or claim 2, comprising means to set the timing of the selected scrambling code relative to the received signal on the basis of timing information forming part of the neighbouring cell list.","4.A method of classifying a ray received at a code division multiple access receiver, the method comprising: receiving a neighbouring cell list comprising at least information concerning scrambling codes used by at least one neighbouring cell, selecting a neighbouring cell from the neighbouring cell list; determining whether an unclassified ray has a timing characteristic falling within a predetermined range of a timing characteristic of a ray classified to the selected cell; and in the event that the timing characteristic of the unclassified ray fails within said predetermined range, mixing a received signal corresponding to the unclassified ray with a scrambling code used by the selected cell and examining the mixed signal to determine if the ray originates from the given neighbouring cell.","5.A method according to claim 4, further comprising setting the timing of the scrambling code relative to the received signal on the basis of timing information forming part of the neighbouring cell list.","6.A method of classifying a ray received at a code division multiple access receiver, the method comprising: (a) receiving a neighbouring cell list comprising at least information concerning scrambling codes used by at least one neighbouring cell; (b) selecting a cell from the neighbouring cell list; (c) determining whether an unclassified ray has a timing reference falling within a predetermined range of a timing reference of a ray which has been classified to the selected neighbouring cell; and (d) in the event that the timing reference is with said predetermined range, correlating a received signal corresponding to the unclassified ray with a scrambling code used by the selected neighbouring cell thereby to determine whether the received signal originates from the selected neighbouring cell.","7.A method according to claim 6, wherein in the event that the timing reference is outside said predetermined range, the method further comprises selecting a different cell from the neighbouring cell list and repeating steps (c) and (d).","8.A method according to claim 6 or claim 7, wherein step (c) comprises determining whether a synchronisation signal of the unclassified ray falls within a predetermined range of a synchronisation signal of the classified ray, the synchronisation signals repeating for each chip of the classified and unclassified ray.","9.A method according to any of claims 6 to 8, wherein step (d) comprises setting the timing of the scrambling code relative to the received signal on the basis of timing information forming part of the neighbouring cell list, the relative timing being modified and step (d) repeated a predetermined number of times if the correlation indicates that the received signal does not originate from the selected neighbouring cell."],["This invention relates to apparatus for, and methods of, classifying a ray received at a code division multiple access receiver.","The proposed universal mobile telephone system (UMTS) will use code division multiple access (CDMA) to communicate on both uplink and downlink communications between node Bs and user equipment (UE).","Downlink communications (node B to UE) will have a scrambling code, specific to the particular node B, modulated thereon.","However, each slot of each frame (there will be fifteen slots in each frame) of downlink communications is proposed to commence with a 256 chip universal word, which is not modulated with the scrambling code.","This universal word is the same for every slot of every frame, and its repeated transmission is hereafter referred to as the primary synchronisation channel (PSCH).","UEs can detect node Bs, or cells, by detecting the PSCH without any knowledge of the identity or location of the node B, its scrambling code or its timing.","However, all the information that can be gleaned from the PSCH is the slot timing.","Frame timing, node B identity and scrambling code information is not available from the PSCH channel.","It is proposed to transmit a secondary synchronisation channel (SSCH), on which a sequence of secondary synchronisation codes (SSCs) are transmitted.","The SSCs are transmitted for the first 256 chips of the fifteen slots in a frame.","The sixteen different SSCs are constructed from position wise multiplication of a Hadamard sequence with a given sequence Z.","The SSCs are orthogonal to the primary synchronisation code (PSC) and are almost orthogonal to the primary scrambling codes.","There are 64 different SSC sequences proposed to be used in UMTS.","A UE will be able, for a ray corresponding to a signal received from a cell, to detect the SSC sequences of a received signal corresponding to the ray for fifteen consecutive slots, and from this determine the scrambling code group and the frame timing using a look-up table.","The received signal will then be correlated against each of eight scrambling codes which form the scrambling code group, and the scrambling code identified by detecting a high degree of correlation from one of the resulting signals.","Determination of the scrambling code and frame and slot timing is known as ray classification.","This method of ray classification, hereafter termed blind ray classification since it uses no knowledge of the cells, is slow since data must be accumulated over at least one frame.","It also requires a relatively large amount of processor time, because the look-up table is large, which is undesirable in terms of power consumption and in terms of maximising processor sharing flexibility.","This method is repeated for each ray which is required to be classified, and is indicated schematically in FIG.","1.Referring to FIG.","1, the method 100 comprises obtaining frame timing and code group information at step 101, and obtaining the scrambling code at step 102.Steps 103 and 104 cause steps 101 and 102 to be repeated for each unclassified ray.","It is proposed for each cell to broadcast a neighbouring cell list on a broadcast channel (BCH).","The neighbouring cell list will contain the identity of the scrambling code and the approximate slot timing (to within one slot) of each cell which is in the area surrounding the cell transmitting the BCH.","Implicit in this information is the scrambling code group.","The BCH channel will have a low data rate and carry large amounts of data, so the neighbouring cell list is not expected to be repeated very often, for example once every five seconds or so.","It is proposed at page 144 of “WCDMA for UMTS” by Antti Toskala and Harri Holma, ISBN 0 471 720518, to assist the blind ray classification method using information concerning the relative timing between cells.","In accordance with a first aspect of the invention, there is provided apparatus for classifying a ray received at a code division multiple access receiver, the apparatus comprising: means for receiving a neighbouring cell list comprising at least information concerning scrambling codes used by at least one neighbouring cell; means for determining whether a received unclassified ray has a timing characteristic falling within a predetermined range of a timing characteristic of a ray which has been classified to a selected neighbouring cell; means for mixing the signal corresponding to the received unclassified ray with a scrambling code used by said selected neighbouring cell thereby to provide a mixed signals the scrambling code being received from the neighbouring cell list; and means for examining the mixed signal to determine if the ray originates from said selected neighbouring cell, the mixing and examining means being arranged to operate only if it is determined that the timing characteristic of the unclassified ray falls within the predetermined range.","According to a second aspect of the invention, there is provided a method of classifying a ray received at a code division multiple access receiver, the method comprising: receiving a neighbouring cell list comprising at least information concerning scrambling codes used by at least one neighbouring cell; selecting a neighbouring cell from the neighbouring cell list; determining whether an unclassified ray has a timing characteristic falling within a predetermined range of a timing characteristic of a ray classified to the selected cell; and in the event that the timing characteristic of the unclassified ray falls within said predetermined range, mixing a received signal corresponding to the unclassified ray with a scrambling code used by the selected cell and examining the mixed signal to determine if the ray originates from the given neighbouring cell.","According to a third aspect of the invention, there is provided a method of classifying a ray received at a code division multiple access receiver, the method comprising: (a) receiving a neighbouring cell list comprising at least information concerning scrambling codes used by at least one neighbouring cell; (b) selecting a cell from the neighbouring cell list; (c) determining whether an unclassified ray has a timing reference filling within a predetermined range of a timing reference of a ray which has been classified to the selected neighbouring cell; and (d) in the event that the timing reference is within said predetermined range, correlating a received signal corresponding to the unclassified ray with a scrambling code used by the selected neighbouring cell thereby to determine whether the received signal originates from the selected neighbouring cell.","The following statements do not define the invention but are nevertheless useful for understanding the invention.","There may be provided apparatus for classifying a ray received at a code division multiple access receiver, the apparatus comprising: means for receiving a neighbouring cell list comprising at least information concerning scrambling codes used by, and timing information concerning at least one neighboring cell; means for selecting a cell from the neighbouring cell list, the cell having associated therewith one or more scrambling codes; means for determining whether the number of cells in the neighbouring cell list exceeds a threshold; means responsive to a positive determination for attempting to classify the ray to the selected cell without utilising means for mixing a received signal with a scrambling code; and means responsive to a negative determination for mixing a received signal with a scrambling code used by the selected cell to provide a mixed signal, and for examining the mixed signal to determine if the ray originates from the cell.","There is also described a method of classifying a ray received at a code division multiple access receiver, the method comprising: receiving a neighbouring cell list comprising at least information concerning scrambling codes used by and timing information concerning at least one neighbouring cell; selecting a cell from the neighbouring cell list, the cell having associated therewith one or more scrambling codes; determining whether the number of cells in the neighbouring cell list exceeds a threshold; in response to a positive determination, attempting to classify the ray to the selected cell using scrambling code group information and without nixing a received signal with a scrambling code; and in response to a negative determination, mixing a received signal with a scrambling code used by the selected cell to provide a mixed signal, and examining the mixed signal to determine if the ray originates from the cell.","There is also described a method of classifying a ray received at a code division multiple access receiver, the method comprising: determining if a timing characteristic of the ray falls within a predetermined range of a timing characteristic of a ray classified to a given cell; in response to a positive determination, mixing a received signal with a scrambling code used by the given cell to provide a mixed signal; and examining the mixed signal to determine if the ray originates from the given cell.","There is also described apparatus for classifying a ray received at a code division multiple access receiver, the apparatus comprising: means for determining if a timing characteristic of the ray falls within a predetermined range of a timing characteristic of a ray classified to a given cell; means responsive to a positive determination for mixing a received signal with a scrambling code used by the given cell to provide a mixed signal; and means for examining the mixed signal to determine if the ray originates from the given cell.","There is also described apparatus for classifying a ray received at a code division multiple access receiver, the apparatus comprising: means for receiving a neighbouring cell list comprising, for each of a plurality of cells, at least timing information and one of a) cell synchronisation code information, and b) information from which cell synchronisation code information can be derived; means for deriving for at least one cell, if necessary, cell synchronisation code information from the neighbouring cell list; a demodulator for demodulating a received synchronisation code from the received ray; means for determining a possible relative phase of a cell synchronisation code, derived from the cell synchronisation code information, for a given cell to the received synchronisation code on the basis of the timing information; a comparator for comparing at least part of the received synchronisation code with at least a part of the cell synchronisation code from the given cell at the relative phase; and means for determining, on the basis of an output of the comparator, whether the received ray originates from the given cell.","There is also described a method of classifying a ray received at a code division multiple access receiver, the method comprising: receiving a neighbouring cell list, comprising for each of a plurality of cells at least timing information and one of a) cell synchronisation code information, and b) information from which cell synchronisation code information can be derived; deriving for at least one cell, if necessary, cell synchronisation code information from the neighbouring cell list; demodulating a received synchronisation code from the received ray; determining a possible relative phase of a cell synchronisation code, derived from the cell synchronisation code information, for a given cell to the received synchronisation code on the basis of the timing information; comparing at least a part of the received synchronisation code with at least a part of the cell synchronisation code for the given cell at the relative phase; determining, on the basis of the result of the comparison, whether the received ray originates from the given cell.","Embodiments of the present invention will now be described, by way of example only; with reference to the accompanying drawings, of which: FIG.","1 is a flow chart illustrating a first, prior art, ray classification method; FIGS.","2 and 3 are flow charts illustrating second and third ray classification methods which are useful for understanding the invention; FIG.","4 is a flow chart illustrating a ray classification method in accordance with the invention; and FIG.","5 is a flow chart illustrating a ray classification method which is useful for understanding the invention.","Referring to FIG.","2, a second ray classification method 200 commences with step 201, which comprises obtaining the SSCH Hadamard code sequences received for a candidate ray in a conventional manner, i.e.","by demodulation of the received signal.","In step 202, the neighbouring cell list received from a cell is examined, and the scrambling code group and slot time are obtained for a cell selected from that list.","In step 203, the obtained sequence of SSCs corresponding to the ray is separately compared against the SSCs for the code group of the scrambling code of the selected cell at the central timing offset and at timing offsets one slot either side of the central timing offset.","If this does not result in classification of the ray, steps 204 and 205 cause the procedure to be repeated for a different cell.","This is performed for each of the cells identified in the neighbouring cell list.","The second method 200 is substantially quicker than the blind ray classification method described above with reference to FIG.","1.Once the SSC sequence has been determined, the blind classification search requires checking the sequence against 64 code groups at fifteen different frame timings, which amounts to 960 checks.","Clearly, far fewer checks are required for each cell in the neighbouring cell list using the second method 200.Also, since the number of cells against which a signal is checked is reduced, the second method 200 is more likely to result in correct classification than the blind method.","In a modification of the second method 200, extra confidence in the cell classification is provided.","Here, a CPICH check is performed at step 204 only when the SSCH search step 203 yields positive result, to confirm that the scrambling code of the selected cell is the same as that used to modulate the received signal.","This CPICH check, put simply, involves correlating a received signal with the scrambling code used by the selected cell, and examining the result.","The relative timing of the two signals is set to isolate the unclassified ray using the detected commencement of PSCH bursts to identify the start of a slot, and aligning the scrambling code accordingly.","Since CPICH channels carry a known data sequence—it is currently anticipated to be all +1s—it is a simple matter to determine whether the scrambling code corresponding to the selected cell is the same as and is synchronised with the scrambling code on the received signal.","Since the frame timing of the ray classified to the selected cell is known, the correlation need only be performed at one of the fifteen possible slot positions.","It will be appreciated that in this embodiment the sequence of SSCs is determined with reference to a look-up table, which relates SSCH sequences to scrambling code groups, from the identification of the scrambling code transmitted as part of the neighbouring cell list.","In an alternative embodiment (not shown) the neighbouring cell list includes the sequence of SSCs, or information from which the sequence of SSCs may be determined.","Such information may be, for example, the identity of the scrambling code group or similar.","Referring now to FIG.","3, the third method 300 starts with step 301 by selecting the unclassified ray which has the highest power.","Step 302 then determines whether the unclassified ray has been checked against all classified rays.","On first performing the method, the answer to step 302 necessarily is no, so step 303 selects one cell from which a ray has been classified.","It is a pre-condition of the third method 300 that at least one ray has been classified, although the method performs better the larger the number of cells from which rays have been classified.","At step 304, the rays classified to the selected cell are examined in turn to determine if one of the classified rays has a timing characteristic which falls within 80 chips of the selected unclassified ray.","In this embodiment, the timing of the start of PSCH bursts is used to determine the relative timing of the unclassified ray and the classified rays, although other references could be used instead.","80 chips correspond to 21 μs or 6.25 km.","If there is a negative determination, the method returns to step 302.If it is determined in step 302 that the unclassified ray has not been checked against all classified rays, step 303 selects a different cell for use by step 304.If step 304 yields a positive determination, step 305 performs a CPICH check using the scrambling code associated with the selected cell.","This CPICH check, put simply, involves correlating a received signal with the scrambling code used by the selected cell, and examining the result.","The relative timing of the two signals is set to isolate the unclassified ray using the detected commencement of PSCH bursts to identify the start of a slot, and aligning the scrambling code accordingly.","Since CPICH channels carry a known data sequence—it is currently anticipated to be all +1s—it is a simple matter to determine whether the scrambling code corresponding to the selected cell is the same as and is synchronised with the scrambling code on the received signal.","Since the frame timing of the ray classified to the selected cell is known, the correlation need only be performed at one of the fifteen possible slot positions.","Step 306 detects whether the CPICH check produced a positive result, and returns to step 302 or advances to step 307 accordingly.","The ray is classified according to scrambling code and frame timing at step 307, then step 308 determines if there are any more unclassified rays.","If there are more unclassified rays, the third method 300 returns to step 301.Otherwise, the method ends.","When step 302 determines that an unclassified ray has been checked against all classified rays, steps 309 and 310 are invoked to classify the ray.","Steps 309 and 310 constitute a blind ray classification, as is described above with reference to FIG.","1, after which the ray is classified in step 307.The third method results in ray classification in a shorter time than the blind ray classification method described above, since with the third method it is not necessary to decode the SSCH for fifteen consecutive slots, which equates to 36,096 chips.","Performing a CPICH check, on the other hand, can require only the first 512 chips of a slot.","The third method also requires less processor time, since the complexity of an SSCH search and subsequent code group identification is significantly higher than the complexity of a single CPICH check.","Furthermore, since the CPICH channel is transmitted at a higher power level than the PSCH and SSCH channels, the third method is more likely to classify a ray to its cell than the blind ray classification method or the second method described above.","Numerous variations on the third method 300 are possible.","For example, a signal corresponding to a candidate ray could be checked against a channel other than the CPICH channel.","The broadcast channel (BCH), for example is suitable for this, since data modulated onto this channel at a low rate and the channel is modulated with a known OVSF code (OVSF code number 1).","CPICH checking is preferred since the CPICH channel is expected to be transmitted at a higher power level than the BCH channel.","Using the results from two channels, such as a BCH channel and a CPICH channel, checked simultaneously or sequentially, provides an increased probability of ray classification if the ray does originate from the cell being checked against.","The third method may be implemented in hardware, in software or in a combination of hardware and software.","Referring now to FIG.","4, the method in accordance with the invention, and hereafter referred to as the fourth method 400 starts with a number of unclassified rays, arranged in descending order according to power, and a number of classified rays.","A neighbouring cell list is decoded from the BCH channel received from a cell which the UE is listening to.","It is not necessary for the UE to be in communication with a node B, although this is likely to be the case in practise.","The candidate ray having the highest power is selected, and a cell is then selected from the neighbouring cell list in step 401.It is then determined whether or not one of the classified rays originates from the cell selected in step 402.If the answer is affirmative, step 403 examines the relative timing of the strongest ray which has been classified to the selected cell to the candidate ray, and determines whether the two rays are separated by more than 80 chips of the scrambling code, which equates to 21 μs or 6.25 km.","In this embodiment, the timing of the start of PSCH bursts is used to determine the relative timing of the unclassified ray to the classified ray, although other references could be used instead.","If the answer to step 403 is affirmative, it is assumed that the candidate ray is not from the selected cell, and the method 400 proceeds to step 404.If in step 402 it is determined that no rays from the selected cell have been classified, a CPICH check is performed on the candidate ray in stop 405 with the scrambling code being one slot earlier than the expected timing of the scrambling code.","The CPICH check put simply involves correlating a received signal with the scrambling code used by the selected cell, and examining the result.","The relative timing of the two signals is set to isolate the unclassified ray using the detected commencement of PSCH bursts as the start of a slot, and aligning the scrambling code accordingly.","Since CPICH channels carry a known data sequence—it is currently anticipated to be all +1s—it is a simple matter to determine whether the scrambling code corresponding to the selected cell is the same as and is synchronised with the scrambling code on the received signal.","If the result of step 405 is affirmative, i.e.","the scrambling code and its timing is correct, then the candidate ray is classified according to that code and that frame timing in step 406, and the method 400 then proceeds to the end.","If the result of step 405 is negative, then a CPICH check is performed in step 407 with relative timing equal to that expected.","If in step 408 it is determined that the scrambling code and its timing is correct, then the candidate ray is classified, and the method 400 proceeds to the end.","Otherwise, a third CPICH check is performed in step 409 with the scrambling code delayed one slot compared to the expected timing of the scrambling code.","If the result is affirmative, the ray is classified accordingly in step 410, and the method proceeds to the end.","Otherwise, it is determined in step 404 if there are any more cells in the neighbouring cell list which have not been tried for the candidate ray.","If there are more cells, the method is recommenced at step 401 where a new cell is selected from the neighbouring cell list.","If there are no more cells, then the method proceeds to the end, and the candidate ray requires classification by another method if it is to be classified.","The fourth method results in ray classification in a shorter time than the blind ray classification method described above, since with the fourth method it is not necessary to decode the SSCH for fifteen consecutive slots, which equates to 36,096 chips.","Performing a CPICH check, on the other hand, can require only the first 512 chips of a slot.","The fourth method also requires less processor time, since the complexity of an SSCH search and subsequent code group identification is significantly higher than the complexity of a CPICH check.","Furthermore, since the CPICH channel is transmitted at a higher power level than the PSCH and SSCH channels, the fourth method is more likely to classify a ray to its cell than the blind ray classification method or the second method described above.","Numerous variations on the fourth method 400 are possible.","For example, a signal corresponding to a candidate ray could be checked against a channel other than the CPICH channel.","The BCH channel, for example, is suitable for this since data modulated onto this channel at a low data rate and the channel is modulated with a known OVSF code (OVSF code number 1).","CPICH checking is preferred since the CPICH channel is expected to be transmitted at a higher power level than the BCH channel.","Using the results from two channels, such as a BCH channel and a CPICH channel, checked simultaneously or sequentially provides an increased probability of ray classification if the ray does originate from the cell being checked against.","If it is not possible to identify the scrambling code used by a cell from the neighbouring cell list, the method 400 may be required to be repeated using different scrambling codes.","If the neighbouring cell list does not contain timing information, then the method 400 may need to be modified to allow CPICH checks at each of the fifteen possible relative timings.","The fourth method may be implemented in hardware, in software or in a combination of hardware and software.","FIG.","5 is a flow chart illustrating a flier ray classification method.","Referring to FIG.","5, the method 500 commence, then selects at step 501 the strongest unclassified ray.","On subsequent runs, the step 501 comprises selecting the next strongest unclassified ray.","At step 502, it is determined whether any rays have so far been classified.","If the answer is yes, then the method 500 proceeds to step 503.If the answer is no, which may occur for example on power-up of a mobile telephone, step 504 causes a blind ray classification check to be made on the ray.","Blind ray classification is described above with reference to FIG.","1.If the ray is not able to be classified in this way, step 506 asks if there are any more unclassified rays, and returns to step 501 or ends the method 500 accordingly.","If the ray is classified, the classification data is stored at step 507 before the method proceeds to step 506.At step 503, the attempt is made to classify the ray using steps 302, 303, 304, 305 and 306 of the third method 300, described above with reference to FIG.","3.If in step 508 it is determined that the ray was classified by this, then the method 500 proceeds to step 507, where the classification data is stored.","Otherwise, the method 500 proceeds to step 509.At step 509, the method 500 detects the number of cells that appear in a neighbouring cell list which is received from a cell over a BCH channel.","The number of neighbouring cells is compared to a threshold N, and the method 500 proceeds to step 510 if the threshold is exceeded, or proceeds to step 511 otherwise.","Step 510 comprises the second method 200, described above with reference to FIG.","2.Step 511 comprises the fourth method 400 described above with reference to FIG.","4.If the ray becomes classified by whichever of steps 510 and 511 were implemented, then step 512 causes the method 500 to pass to step 507, where the classification data is stored.","Otherwise, the method passes to step 504 where blind classification of the ray is attempted.","The value of the threshold N is hardwired into the apparatus which implements the method.","This invention may be implemented in hardware, software or in a combination of hardware and software.","The value of the threshold is selected dependent on a number of factors including the required speed of ray classification, processor speed and processor availability.","It will be appreciated that the fourth method 400 requires less time to check a ray than the second method 200, unless there are a large number of cells in the neighbouring cell list since less time is required to acquire the necessary data.","Also, the amount of processing required to implement the fourth method 400 increases greatly as the number of cells in the neighbouring cell list increases, although this is not true of the second method.","In another embodiment, the value of the threshold is dynamically adjustable by a controller (not shown)."]],"string":"[\n [\n \"Method and apparatus for fast w-cdma acquisition\",\n \"Apparatus for classifying a ray is disclosed, wherein the ray is received at a code division multiple access receiver.\",\n \"The apparatus comprises: means for receiving a neighbouring cell list comprising at least information concerning scrambling codes used by at least one neighbouring cell; means for selecting a scrambling code used by a given neighbouring cell; means for mixing a received signal with the selected scrambling code to provide a mixed signal; and means for examining the mixed signal to determine if the ray originates from the given neighbouring cell.\"\n ],\n [\n \"1.Apparatus for classifying a ray received at a code division multiple access receiver, the apparatus comprising; means for receiving a neighbouring cell list comprising at least information concerning scrambling codes used by at least one neighbouring cell; means for determining whether a received unclassified ray has a timing characteristic falling within a predetermined range of a timing characteristic of a ray which has been classified to a selected neighbouring cell; means for mixing the signal corresponding to the received unclassified ray with a scrambling code used by said selected neighbouring cell thereby to provide a mixed signal, the scrambling code being received from the neighbouring cell list; and means for examining the mixed signal to determine if the ray originates from said selected neighbouring cell, the mixing and examining means being arranged to operate only if it is determined that the timing characteristic of the unclassified ray falls within the predetermined range.\",\n \"2.Apparatus according to claim 1, further comprising means arranged to select a different cell from the neighbouring cell list in the event that it is determined that the timing characteristic of the unclassified ray falls outside the predetermined range.\",\n \"3.Apparatus according to claim 1 or claim 2, comprising means to set the timing of the selected scrambling code relative to the received signal on the basis of timing information forming part of the neighbouring cell list.\",\n \"4.A method of classifying a ray received at a code division multiple access receiver, the method comprising: receiving a neighbouring cell list comprising at least information concerning scrambling codes used by at least one neighbouring cell, selecting a neighbouring cell from the neighbouring cell list; determining whether an unclassified ray has a timing characteristic falling within a predetermined range of a timing characteristic of a ray classified to the selected cell; and in the event that the timing characteristic of the unclassified ray fails within said predetermined range, mixing a received signal corresponding to the unclassified ray with a scrambling code used by the selected cell and examining the mixed signal to determine if the ray originates from the given neighbouring cell.\",\n \"5.A method according to claim 4, further comprising setting the timing of the scrambling code relative to the received signal on the basis of timing information forming part of the neighbouring cell list.\",\n \"6.A method of classifying a ray received at a code division multiple access receiver, the method comprising: (a) receiving a neighbouring cell list comprising at least information concerning scrambling codes used by at least one neighbouring cell; (b) selecting a cell from the neighbouring cell list; (c) determining whether an unclassified ray has a timing reference falling within a predetermined range of a timing reference of a ray which has been classified to the selected neighbouring cell; and (d) in the event that the timing reference is with said predetermined range, correlating a received signal corresponding to the unclassified ray with a scrambling code used by the selected neighbouring cell thereby to determine whether the received signal originates from the selected neighbouring cell.\",\n \"7.A method according to claim 6, wherein in the event that the timing reference is outside said predetermined range, the method further comprises selecting a different cell from the neighbouring cell list and repeating steps (c) and (d).\",\n \"8.A method according to claim 6 or claim 7, wherein step (c) comprises determining whether a synchronisation signal of the unclassified ray falls within a predetermined range of a synchronisation signal of the classified ray, the synchronisation signals repeating for each chip of the classified and unclassified ray.\",\n \"9.A method according to any of claims 6 to 8, wherein step (d) comprises setting the timing of the scrambling code relative to the received signal on the basis of timing information forming part of the neighbouring cell list, the relative timing being modified and step (d) repeated a predetermined number of times if the correlation indicates that the received signal does not originate from the selected neighbouring cell.\"\n ],\n [\n \"This invention relates to apparatus for, and methods of, classifying a ray received at a code division multiple access receiver.\",\n \"The proposed universal mobile telephone system (UMTS) will use code division multiple access (CDMA) to communicate on both uplink and downlink communications between node Bs and user equipment (UE).\",\n \"Downlink communications (node B to UE) will have a scrambling code, specific to the particular node B, modulated thereon.\",\n \"However, each slot of each frame (there will be fifteen slots in each frame) of downlink communications is proposed to commence with a 256 chip universal word, which is not modulated with the scrambling code.\",\n \"This universal word is the same for every slot of every frame, and its repeated transmission is hereafter referred to as the primary synchronisation channel (PSCH).\",\n \"UEs can detect node Bs, or cells, by detecting the PSCH without any knowledge of the identity or location of the node B, its scrambling code or its timing.\",\n \"However, all the information that can be gleaned from the PSCH is the slot timing.\",\n \"Frame timing, node B identity and scrambling code information is not available from the PSCH channel.\",\n \"It is proposed to transmit a secondary synchronisation channel (SSCH), on which a sequence of secondary synchronisation codes (SSCs) are transmitted.\",\n \"The SSCs are transmitted for the first 256 chips of the fifteen slots in a frame.\",\n \"The sixteen different SSCs are constructed from position wise multiplication of a Hadamard sequence with a given sequence Z.\",\n \"The SSCs are orthogonal to the primary synchronisation code (PSC) and are almost orthogonal to the primary scrambling codes.\",\n \"There are 64 different SSC sequences proposed to be used in UMTS.\",\n \"A UE will be able, for a ray corresponding to a signal received from a cell, to detect the SSC sequences of a received signal corresponding to the ray for fifteen consecutive slots, and from this determine the scrambling code group and the frame timing using a look-up table.\",\n \"The received signal will then be correlated against each of eight scrambling codes which form the scrambling code group, and the scrambling code identified by detecting a high degree of correlation from one of the resulting signals.\",\n \"Determination of the scrambling code and frame and slot timing is known as ray classification.\",\n \"This method of ray classification, hereafter termed blind ray classification since it uses no knowledge of the cells, is slow since data must be accumulated over at least one frame.\",\n \"It also requires a relatively large amount of processor time, because the look-up table is large, which is undesirable in terms of power consumption and in terms of maximising processor sharing flexibility.\",\n \"This method is repeated for each ray which is required to be classified, and is indicated schematically in FIG.\",\n \"1.Referring to FIG.\",\n \"1, the method 100 comprises obtaining frame timing and code group information at step 101, and obtaining the scrambling code at step 102.Steps 103 and 104 cause steps 101 and 102 to be repeated for each unclassified ray.\",\n \"It is proposed for each cell to broadcast a neighbouring cell list on a broadcast channel (BCH).\",\n \"The neighbouring cell list will contain the identity of the scrambling code and the approximate slot timing (to within one slot) of each cell which is in the area surrounding the cell transmitting the BCH.\",\n \"Implicit in this information is the scrambling code group.\",\n \"The BCH channel will have a low data rate and carry large amounts of data, so the neighbouring cell list is not expected to be repeated very often, for example once every five seconds or so.\",\n \"It is proposed at page 144 of “WCDMA for UMTS” by Antti Toskala and Harri Holma, ISBN 0 471 720518, to assist the blind ray classification method using information concerning the relative timing between cells.\",\n \"In accordance with a first aspect of the invention, there is provided apparatus for classifying a ray received at a code division multiple access receiver, the apparatus comprising: means for receiving a neighbouring cell list comprising at least information concerning scrambling codes used by at least one neighbouring cell; means for determining whether a received unclassified ray has a timing characteristic falling within a predetermined range of a timing characteristic of a ray which has been classified to a selected neighbouring cell; means for mixing the signal corresponding to the received unclassified ray with a scrambling code used by said selected neighbouring cell thereby to provide a mixed signals the scrambling code being received from the neighbouring cell list; and means for examining the mixed signal to determine if the ray originates from said selected neighbouring cell, the mixing and examining means being arranged to operate only if it is determined that the timing characteristic of the unclassified ray falls within the predetermined range.\",\n \"According to a second aspect of the invention, there is provided a method of classifying a ray received at a code division multiple access receiver, the method comprising: receiving a neighbouring cell list comprising at least information concerning scrambling codes used by at least one neighbouring cell; selecting a neighbouring cell from the neighbouring cell list; determining whether an unclassified ray has a timing characteristic falling within a predetermined range of a timing characteristic of a ray classified to the selected cell; and in the event that the timing characteristic of the unclassified ray falls within said predetermined range, mixing a received signal corresponding to the unclassified ray with a scrambling code used by the selected cell and examining the mixed signal to determine if the ray originates from the given neighbouring cell.\",\n \"According to a third aspect of the invention, there is provided a method of classifying a ray received at a code division multiple access receiver, the method comprising: (a) receiving a neighbouring cell list comprising at least information concerning scrambling codes used by at least one neighbouring cell; (b) selecting a cell from the neighbouring cell list; (c) determining whether an unclassified ray has a timing reference filling within a predetermined range of a timing reference of a ray which has been classified to the selected neighbouring cell; and (d) in the event that the timing reference is within said predetermined range, correlating a received signal corresponding to the unclassified ray with a scrambling code used by the selected neighbouring cell thereby to determine whether the received signal originates from the selected neighbouring cell.\",\n \"The following statements do not define the invention but are nevertheless useful for understanding the invention.\",\n \"There may be provided apparatus for classifying a ray received at a code division multiple access receiver, the apparatus comprising: means for receiving a neighbouring cell list comprising at least information concerning scrambling codes used by, and timing information concerning at least one neighboring cell; means for selecting a cell from the neighbouring cell list, the cell having associated therewith one or more scrambling codes; means for determining whether the number of cells in the neighbouring cell list exceeds a threshold; means responsive to a positive determination for attempting to classify the ray to the selected cell without utilising means for mixing a received signal with a scrambling code; and means responsive to a negative determination for mixing a received signal with a scrambling code used by the selected cell to provide a mixed signal, and for examining the mixed signal to determine if the ray originates from the cell.\",\n \"There is also described a method of classifying a ray received at a code division multiple access receiver, the method comprising: receiving a neighbouring cell list comprising at least information concerning scrambling codes used by and timing information concerning at least one neighbouring cell; selecting a cell from the neighbouring cell list, the cell having associated therewith one or more scrambling codes; determining whether the number of cells in the neighbouring cell list exceeds a threshold; in response to a positive determination, attempting to classify the ray to the selected cell using scrambling code group information and without nixing a received signal with a scrambling code; and in response to a negative determination, mixing a received signal with a scrambling code used by the selected cell to provide a mixed signal, and examining the mixed signal to determine if the ray originates from the cell.\",\n \"There is also described a method of classifying a ray received at a code division multiple access receiver, the method comprising: determining if a timing characteristic of the ray falls within a predetermined range of a timing characteristic of a ray classified to a given cell; in response to a positive determination, mixing a received signal with a scrambling code used by the given cell to provide a mixed signal; and examining the mixed signal to determine if the ray originates from the given cell.\",\n \"There is also described apparatus for classifying a ray received at a code division multiple access receiver, the apparatus comprising: means for determining if a timing characteristic of the ray falls within a predetermined range of a timing characteristic of a ray classified to a given cell; means responsive to a positive determination for mixing a received signal with a scrambling code used by the given cell to provide a mixed signal; and means for examining the mixed signal to determine if the ray originates from the given cell.\",\n \"There is also described apparatus for classifying a ray received at a code division multiple access receiver, the apparatus comprising: means for receiving a neighbouring cell list comprising, for each of a plurality of cells, at least timing information and one of a) cell synchronisation code information, and b) information from which cell synchronisation code information can be derived; means for deriving for at least one cell, if necessary, cell synchronisation code information from the neighbouring cell list; a demodulator for demodulating a received synchronisation code from the received ray; means for determining a possible relative phase of a cell synchronisation code, derived from the cell synchronisation code information, for a given cell to the received synchronisation code on the basis of the timing information; a comparator for comparing at least part of the received synchronisation code with at least a part of the cell synchronisation code from the given cell at the relative phase; and means for determining, on the basis of an output of the comparator, whether the received ray originates from the given cell.\",\n \"There is also described a method of classifying a ray received at a code division multiple access receiver, the method comprising: receiving a neighbouring cell list, comprising for each of a plurality of cells at least timing information and one of a) cell synchronisation code information, and b) information from which cell synchronisation code information can be derived; deriving for at least one cell, if necessary, cell synchronisation code information from the neighbouring cell list; demodulating a received synchronisation code from the received ray; determining a possible relative phase of a cell synchronisation code, derived from the cell synchronisation code information, for a given cell to the received synchronisation code on the basis of the timing information; comparing at least a part of the received synchronisation code with at least a part of the cell synchronisation code for the given cell at the relative phase; determining, on the basis of the result of the comparison, whether the received ray originates from the given cell.\",\n \"Embodiments of the present invention will now be described, by way of example only; with reference to the accompanying drawings, of which: FIG.\",\n \"1 is a flow chart illustrating a first, prior art, ray classification method; FIGS.\",\n \"2 and 3 are flow charts illustrating second and third ray classification methods which are useful for understanding the invention; FIG.\",\n \"4 is a flow chart illustrating a ray classification method in accordance with the invention; and FIG.\",\n \"5 is a flow chart illustrating a ray classification method which is useful for understanding the invention.\",\n \"Referring to FIG.\",\n \"2, a second ray classification method 200 commences with step 201, which comprises obtaining the SSCH Hadamard code sequences received for a candidate ray in a conventional manner, i.e.\",\n \"by demodulation of the received signal.\",\n \"In step 202, the neighbouring cell list received from a cell is examined, and the scrambling code group and slot time are obtained for a cell selected from that list.\",\n \"In step 203, the obtained sequence of SSCs corresponding to the ray is separately compared against the SSCs for the code group of the scrambling code of the selected cell at the central timing offset and at timing offsets one slot either side of the central timing offset.\",\n \"If this does not result in classification of the ray, steps 204 and 205 cause the procedure to be repeated for a different cell.\",\n \"This is performed for each of the cells identified in the neighbouring cell list.\",\n \"The second method 200 is substantially quicker than the blind ray classification method described above with reference to FIG.\",\n \"1.Once the SSC sequence has been determined, the blind classification search requires checking the sequence against 64 code groups at fifteen different frame timings, which amounts to 960 checks.\",\n \"Clearly, far fewer checks are required for each cell in the neighbouring cell list using the second method 200.Also, since the number of cells against which a signal is checked is reduced, the second method 200 is more likely to result in correct classification than the blind method.\",\n \"In a modification of the second method 200, extra confidence in the cell classification is provided.\",\n \"Here, a CPICH check is performed at step 204 only when the SSCH search step 203 yields positive result, to confirm that the scrambling code of the selected cell is the same as that used to modulate the received signal.\",\n \"This CPICH check, put simply, involves correlating a received signal with the scrambling code used by the selected cell, and examining the result.\",\n \"The relative timing of the two signals is set to isolate the unclassified ray using the detected commencement of PSCH bursts to identify the start of a slot, and aligning the scrambling code accordingly.\",\n \"Since CPICH channels carry a known data sequence—it is currently anticipated to be all +1s—it is a simple matter to determine whether the scrambling code corresponding to the selected cell is the same as and is synchronised with the scrambling code on the received signal.\",\n \"Since the frame timing of the ray classified to the selected cell is known, the correlation need only be performed at one of the fifteen possible slot positions.\",\n \"It will be appreciated that in this embodiment the sequence of SSCs is determined with reference to a look-up table, which relates SSCH sequences to scrambling code groups, from the identification of the scrambling code transmitted as part of the neighbouring cell list.\",\n \"In an alternative embodiment (not shown) the neighbouring cell list includes the sequence of SSCs, or information from which the sequence of SSCs may be determined.\",\n \"Such information may be, for example, the identity of the scrambling code group or similar.\",\n \"Referring now to FIG.\",\n \"3, the third method 300 starts with step 301 by selecting the unclassified ray which has the highest power.\",\n \"Step 302 then determines whether the unclassified ray has been checked against all classified rays.\",\n \"On first performing the method, the answer to step 302 necessarily is no, so step 303 selects one cell from which a ray has been classified.\",\n \"It is a pre-condition of the third method 300 that at least one ray has been classified, although the method performs better the larger the number of cells from which rays have been classified.\",\n \"At step 304, the rays classified to the selected cell are examined in turn to determine if one of the classified rays has a timing characteristic which falls within 80 chips of the selected unclassified ray.\",\n \"In this embodiment, the timing of the start of PSCH bursts is used to determine the relative timing of the unclassified ray and the classified rays, although other references could be used instead.\",\n \"80 chips correspond to 21 μs or 6.25 km.\",\n \"If there is a negative determination, the method returns to step 302.If it is determined in step 302 that the unclassified ray has not been checked against all classified rays, step 303 selects a different cell for use by step 304.If step 304 yields a positive determination, step 305 performs a CPICH check using the scrambling code associated with the selected cell.\",\n \"This CPICH check, put simply, involves correlating a received signal with the scrambling code used by the selected cell, and examining the result.\",\n \"The relative timing of the two signals is set to isolate the unclassified ray using the detected commencement of PSCH bursts to identify the start of a slot, and aligning the scrambling code accordingly.\",\n \"Since CPICH channels carry a known data sequence—it is currently anticipated to be all +1s—it is a simple matter to determine whether the scrambling code corresponding to the selected cell is the same as and is synchronised with the scrambling code on the received signal.\",\n \"Since the frame timing of the ray classified to the selected cell is known, the correlation need only be performed at one of the fifteen possible slot positions.\",\n \"Step 306 detects whether the CPICH check produced a positive result, and returns to step 302 or advances to step 307 accordingly.\",\n \"The ray is classified according to scrambling code and frame timing at step 307, then step 308 determines if there are any more unclassified rays.\",\n \"If there are more unclassified rays, the third method 300 returns to step 301.Otherwise, the method ends.\",\n \"When step 302 determines that an unclassified ray has been checked against all classified rays, steps 309 and 310 are invoked to classify the ray.\",\n \"Steps 309 and 310 constitute a blind ray classification, as is described above with reference to FIG.\",\n \"1, after which the ray is classified in step 307.The third method results in ray classification in a shorter time than the blind ray classification method described above, since with the third method it is not necessary to decode the SSCH for fifteen consecutive slots, which equates to 36,096 chips.\",\n \"Performing a CPICH check, on the other hand, can require only the first 512 chips of a slot.\",\n \"The third method also requires less processor time, since the complexity of an SSCH search and subsequent code group identification is significantly higher than the complexity of a single CPICH check.\",\n \"Furthermore, since the CPICH channel is transmitted at a higher power level than the PSCH and SSCH channels, the third method is more likely to classify a ray to its cell than the blind ray classification method or the second method described above.\",\n \"Numerous variations on the third method 300 are possible.\",\n \"For example, a signal corresponding to a candidate ray could be checked against a channel other than the CPICH channel.\",\n \"The broadcast channel (BCH), for example is suitable for this, since data modulated onto this channel at a low rate and the channel is modulated with a known OVSF code (OVSF code number 1).\",\n \"CPICH checking is preferred since the CPICH channel is expected to be transmitted at a higher power level than the BCH channel.\",\n \"Using the results from two channels, such as a BCH channel and a CPICH channel, checked simultaneously or sequentially, provides an increased probability of ray classification if the ray does originate from the cell being checked against.\",\n \"The third method may be implemented in hardware, in software or in a combination of hardware and software.\",\n \"Referring now to FIG.\",\n \"4, the method in accordance with the invention, and hereafter referred to as the fourth method 400 starts with a number of unclassified rays, arranged in descending order according to power, and a number of classified rays.\",\n \"A neighbouring cell list is decoded from the BCH channel received from a cell which the UE is listening to.\",\n \"It is not necessary for the UE to be in communication with a node B, although this is likely to be the case in practise.\",\n \"The candidate ray having the highest power is selected, and a cell is then selected from the neighbouring cell list in step 401.It is then determined whether or not one of the classified rays originates from the cell selected in step 402.If the answer is affirmative, step 403 examines the relative timing of the strongest ray which has been classified to the selected cell to the candidate ray, and determines whether the two rays are separated by more than 80 chips of the scrambling code, which equates to 21 μs or 6.25 km.\",\n \"In this embodiment, the timing of the start of PSCH bursts is used to determine the relative timing of the unclassified ray to the classified ray, although other references could be used instead.\",\n \"If the answer to step 403 is affirmative, it is assumed that the candidate ray is not from the selected cell, and the method 400 proceeds to step 404.If in step 402 it is determined that no rays from the selected cell have been classified, a CPICH check is performed on the candidate ray in stop 405 with the scrambling code being one slot earlier than the expected timing of the scrambling code.\",\n \"The CPICH check put simply involves correlating a received signal with the scrambling code used by the selected cell, and examining the result.\",\n \"The relative timing of the two signals is set to isolate the unclassified ray using the detected commencement of PSCH bursts as the start of a slot, and aligning the scrambling code accordingly.\",\n \"Since CPICH channels carry a known data sequence—it is currently anticipated to be all +1s—it is a simple matter to determine whether the scrambling code corresponding to the selected cell is the same as and is synchronised with the scrambling code on the received signal.\",\n \"If the result of step 405 is affirmative, i.e.\",\n \"the scrambling code and its timing is correct, then the candidate ray is classified according to that code and that frame timing in step 406, and the method 400 then proceeds to the end.\",\n \"If the result of step 405 is negative, then a CPICH check is performed in step 407 with relative timing equal to that expected.\",\n \"If in step 408 it is determined that the scrambling code and its timing is correct, then the candidate ray is classified, and the method 400 proceeds to the end.\",\n \"Otherwise, a third CPICH check is performed in step 409 with the scrambling code delayed one slot compared to the expected timing of the scrambling code.\",\n \"If the result is affirmative, the ray is classified accordingly in step 410, and the method proceeds to the end.\",\n \"Otherwise, it is determined in step 404 if there are any more cells in the neighbouring cell list which have not been tried for the candidate ray.\",\n \"If there are more cells, the method is recommenced at step 401 where a new cell is selected from the neighbouring cell list.\",\n \"If there are no more cells, then the method proceeds to the end, and the candidate ray requires classification by another method if it is to be classified.\",\n \"The fourth method results in ray classification in a shorter time than the blind ray classification method described above, since with the fourth method it is not necessary to decode the SSCH for fifteen consecutive slots, which equates to 36,096 chips.\",\n \"Performing a CPICH check, on the other hand, can require only the first 512 chips of a slot.\",\n \"The fourth method also requires less processor time, since the complexity of an SSCH search and subsequent code group identification is significantly higher than the complexity of a CPICH check.\",\n \"Furthermore, since the CPICH channel is transmitted at a higher power level than the PSCH and SSCH channels, the fourth method is more likely to classify a ray to its cell than the blind ray classification method or the second method described above.\",\n \"Numerous variations on the fourth method 400 are possible.\",\n \"For example, a signal corresponding to a candidate ray could be checked against a channel other than the CPICH channel.\",\n \"The BCH channel, for example, is suitable for this since data modulated onto this channel at a low data rate and the channel is modulated with a known OVSF code (OVSF code number 1).\",\n \"CPICH checking is preferred since the CPICH channel is expected to be transmitted at a higher power level than the BCH channel.\",\n \"Using the results from two channels, such as a BCH channel and a CPICH channel, checked simultaneously or sequentially provides an increased probability of ray classification if the ray does originate from the cell being checked against.\",\n \"If it is not possible to identify the scrambling code used by a cell from the neighbouring cell list, the method 400 may be required to be repeated using different scrambling codes.\",\n \"If the neighbouring cell list does not contain timing information, then the method 400 may need to be modified to allow CPICH checks at each of the fifteen possible relative timings.\",\n \"The fourth method may be implemented in hardware, in software or in a combination of hardware and software.\",\n \"FIG.\",\n \"5 is a flow chart illustrating a flier ray classification method.\",\n \"Referring to FIG.\",\n \"5, the method 500 commence, then selects at step 501 the strongest unclassified ray.\",\n \"On subsequent runs, the step 501 comprises selecting the next strongest unclassified ray.\",\n \"At step 502, it is determined whether any rays have so far been classified.\",\n \"If the answer is yes, then the method 500 proceeds to step 503.If the answer is no, which may occur for example on power-up of a mobile telephone, step 504 causes a blind ray classification check to be made on the ray.\",\n \"Blind ray classification is described above with reference to FIG.\",\n \"1.If the ray is not able to be classified in this way, step 506 asks if there are any more unclassified rays, and returns to step 501 or ends the method 500 accordingly.\",\n \"If the ray is classified, the classification data is stored at step 507 before the method proceeds to step 506.At step 503, the attempt is made to classify the ray using steps 302, 303, 304, 305 and 306 of the third method 300, described above with reference to FIG.\",\n \"3.If in step 508 it is determined that the ray was classified by this, then the method 500 proceeds to step 507, where the classification data is stored.\",\n \"Otherwise, the method 500 proceeds to step 509.At step 509, the method 500 detects the number of cells that appear in a neighbouring cell list which is received from a cell over a BCH channel.\",\n \"The number of neighbouring cells is compared to a threshold N, and the method 500 proceeds to step 510 if the threshold is exceeded, or proceeds to step 511 otherwise.\",\n \"Step 510 comprises the second method 200, described above with reference to FIG.\",\n \"2.Step 511 comprises the fourth method 400 described above with reference to FIG.\",\n \"4.If the ray becomes classified by whichever of steps 510 and 511 were implemented, then step 512 causes the method 500 to pass to step 507, where the classification data is stored.\",\n \"Otherwise, the method passes to step 504 where blind classification of the ray is attempted.\",\n \"The value of the threshold N is hardwired into the apparatus which implements the method.\",\n \"This invention may be implemented in hardware, software or in a combination of hardware and software.\",\n \"The value of the threshold is selected dependent on a number of factors including the required speed of ray classification, processor speed and processor availability.\",\n \"It will be appreciated that the fourth method 400 requires less time to check a ray than the second method 200, unless there are a large number of cells in the neighbouring cell list since less time is required to acquire the necessary data.\",\n \"Also, the amount of processing required to implement the fourth method 400 increases greatly as the number of cells in the neighbouring cell list increases, although this is not true of the second method.\",\n \"In another embodiment, the value of the threshold is dynamically adjustable by a controller (not shown).\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473922"}}},{"rowIdx":765,"cells":{"text":{"kind":"list like","value":[["Endosteal electrode","An implantable tissue-stimulating device comprising an elongate electrode carrier member (11) having a plurality of electrodes thereon.","The electrodes are preferably disposed in a linear array on the carrier member (11) and are adapted to apply a preselected tissue stimulation to the cochlea.","The carrier member (11) is preformed from a resiliently flexible biocompatible silicone and extends from a distal end (12) to a stop member (13).","The carrier member (11) is adapted for intracochlear but extraluminar insertion within the cochlea of an implantee.","In particular, the carrier member (11) is adapted to be implanted in the crevice (21) between the spiral ligament (22) and the endosteum (23) of the lateral wall of the cochlea (20).","This is a quite different location to the normal placement of the cochlear implant electrode array in the scala tympani (24) of the cochlea (20).","The placement of the carrier member (11) is designed to avoid any breach of the internal ducts of the cochlea (20), such as the scala tympani (24) and scala vestibuli (25) so that the normal hydrodynamic behaviour of the cochlea (20) is not affected by any intrusive device.","By preservng the normal hydrodynamic behaviour of the cochlea (20), use of the carrier member (11) maximises the possibility of also preserving any hearing of the implantee that is offered by the cochlea (20).","Use of the device in a system for masking or treating the symptoms of tinnitus is also described."],["1.An implantable tissue-stimulating device having a carrier member having at least one electrode thereon, the carrier member being adapted for intracochlear but extraluminar insertion within the cochlea of an implantee.","2.An implantable tissue-stimulating device for use in the masking or treatment of the symptoms of tinnitus, the device having a carrier member having at least one electrode thereon, the carrier member being adapted for intracochlear but extraluminar insertion within the cochlea of an implantee.","3.An implantable tissue-stimulating device when used in the masking or treatment of the symptoms of tinnitus, the device having a carrier member having at least one electrode thereon, the device being adapted for intracochlear but extraluminar insertion within the cochlea of an implantee.","4.An implantable tissue-stimulating device of claim 1 wherein the carrier member has a body having a plurality of electrodes mounted thereon.","5.An implantable tissue-stimulating device of claim 4 wherein the electrodes are disposed in an array on the carrier member and are adapted to apply a preselected tissue stimulation.","6.An implantable tissue-stimulating device of claim 1 wherein the carrier member is implantable in a crevice between the spiral ligament and the endosteum of the lateral wall of the cochlea.","7.An implantable tissue-stimulating device of claim 6 wherein the carrier member has a maximum length of about 7 mm-10 mm, a width of about 0.6 mm and a thickness no greater than about 0.2 mm and, more preferably, about 0.1 mm.","8.An implantable tissue-stimulating device of claim 6 wherein the carrier member has an inner surface and an outer surface, the inner surface being adapted on implantation to face inwardly into the cochlea.","9.An implantable tissue-stimulating device of claim 8 wherein the inner face has a concave surface and the outer face has a convex surface.","10.An implantable tissue-stimulating device of claim 1 wherein a stop member extends outwardly from the carrier member substantially at right angles to the longitudinal axis of the carrier member.","11.An implantable tissue-stimulating device of claim 10 wherein the stop member has a length of between about 1.5 and 2.0 mm.","12.An implantable tissue-stimulating device of claim 1 wherein the electrodes of the carrier member receive stimulation signals from an implantable stimulator means.","13.An implantable tissue-stimulating device of claim 12 wherein the stimulator means is housed within a housing that also houses a receiver means that is adapted to receive signals from a controller means.","14.An implantable tissue-stimulating device of claim 13 wherein the controller means comprises a processor means adapted to output one or more stimulation regimes to the stimulator.","15.An implantable tissue-stimulating device of claim 14 wherein, where the device is being used to mask or treat the symptoms of tinnitus, the stimulation regime is selected from the group comprising a random continuous sub-threshold stimulation regime, a random continuous supra-threshold stimulation regime, and a random discontinuous supra-threshold stimulation regime.","16.An implantable tissue-stimulating device of claim 14 wherein, where the device is being used to mask or treat the symptoms of tinnitus, the stimulation regime comprises a treatment-on-demand regime.","17.An implantable tissue-stimulating device of claim 16 wherein the controller means further has an activation means that allows the implantee or a third person to activate the processor means when required.","18.An implantable tissue-stimulating device of claim 14 wherein, where the device is being used to provide a hearing sensation, the controller means comprises a speech processor adapted to receive signals output by a microphone and encode the sound detected by the microphone into a sequence of electrical stimuli following given algorithms.","19.An implantable tissue-stimulating device of claim 18 wherein the speech processor is adapted to receive signals from the microphone when external sounds, such as speech, are present and to output a stimulation regime, when no external sounds are present, that is adapted to mask or treat the symptoms of tinnitus.","20.An implantable tissue-stimulating device of claim 14 wherein the controller means is implantable and contained within a hermetically sealed housing or a housing used for the stimulator means.","21.An implantable tissue-stimulating device of claim 20 wherein the controller means has an activation means that provides the implantee or a third person with a means to activate a stimulation regime when required.","22.A method of inserting a tissue-stimulating device as defined in claim 1 into a cochlea of an implantee, the method comprising the steps of: performing a fenestration to access the crevice of the cochlea between the spiral ligament and the endosteum of the lateral wall of the cochlea; placing the device in the crevice; and sealing the fenestration.","23.A method of treating the symptoms of tinnitus comprising inserting a device as defined in claim 1 into a cochlea of an implantee, the method comprising the steps of: performing a fenestration to access the crevice of the cochlea between the spiral ligament and the endosteum of the lateral wall of the cochlea; placing the device in the crevice; and sealing the fenestration."],["<SOH> BACKGROUND OF THE INVENTION <EOH>Hearing loss, which may be due to many different causes, is generally of two types, conductive and sensorineural.","In some cases, a person may have hearing loss of both types.","Of them, conductive hearing loss occurs where the normal mechanical pathways for sound to reach the hair cells in the cochlea are impeded, for example, by damage to the ossicdes.","Conductive hearing loss may often be helped by use of conventional hearing aids, which amplify sound so that acoustic information does reach the cochlea and the hair cells.","In many people who are profoundly deaf, however, the reason for their deafness is sensorineural hearing loss.","This type of hearing loss is due to the absence of, or destruction of, the hair cells in the cochlea which transduce acoustic signals into nerve impulses.","These people are thus unable to derive suitable benefit from conventional hearing aid systems, no matter how loud the acoustic stimulus is made, because there is damage to or absence of the mechanism for nerve impulses to be generated from sound in the normal manner.","It is for this purpose that cochlear implant systems have been developed.","Such systems bypass the hair cells in the cochlea and directly deliver electrical stimulation to the auditory nerve fibres, thereby allowing the brain to perceive a hearing sensation resembling the natural hearing sensation normally delivered to the auditory nerve.","U.S. Pat.","No.","4,532,930, the contents of which are incorporated herein by reference, provides a description of one type of traditional cochlear implant system.","Typically, cochlear implant systems have consisted of essentially two components, an external component commonly referred to as a processor unit and an internal implanted component commonly referred to as a receiver/stimulator unit.","Traditionally, both of these components have cooperated together to provide the sound sensation to a user.","The external component has traditionally consisted of a microphone for detecting sounds, such as speech and environmental sounds, a speech processor that converts speech into a coded signal, a power source such as a battery, and an external transmitter coil.","The coded signal output by the sound processor is transmitted transcutaneously to the implanted receiver/stimulator unit situated within a recess of the temporal bone of the user.","This transcutaneous transmission occurs via the external transmitter coil which is positioned to communicate with an implanted receiver coil provided with the stimulator/receiver unit.","This communication serves two essential purposes, firstly to transcutaneously transmit the coded sound signal and secondly to provide power to the implanted receiver/stimulator unit Conventionally, this link has been in the form of a radio frequency (RF) link, but other such links have been proposed and implemented with varying degrees of success.","The implanted receiver/stimulator unit traditionally includes a receiver coil that receives the coded signal and power from the external processor component, and a stimulator that processes the coded signal and outputs a stimulation signal to an intracochlea electrode assembly which applies the electrical stimulation directly to the auditory nerve producing a hearing sensation corresponding to the original detected sound.","Traditionally, at least the speech processor of the external componentry has been carried on the body of the user, such as in a pocket of the user's clothing, a belt pouch or in a harness, while the microphone has been mounted on a clip mounted behind the ear or on the lapel of the user.","More recently, due in the main to improvements in technology, the physical dimensions of the sound processor have been able to be reduced allowing for the external componentry to be housed in a small unit capable of being worn behind the ear of the user.","This unit allows the microphone, power unit and the sound processor to be housed in a single unit capable of being discretely worn behind the ear, with the external transmitter coil still positioned on the side of the user's head to allow for the transmission of the coded sound signal from the sound processor and power to the implanted stimulator unit.","It is further envisaged that with continual improvements in technology all the traditional external componentry may be implanted in the user.","In such a system all of the speech processing may be performed inside the implanted stimulator unit, via an implanted microphone.","It is known in the art that the cochlea is tonotopically mapped.","In other words, the cochlea can be partitioned into regions, with each region being responsive to signals in a particular frequency range.","This property of the cochlea has been exploited by providing the electrode assembly with an array of electrodes, each electrode being arranged and constructed to deliver a stimulating signal within a preselected frequency range, to the appropriate region within the scala tympani of the cochlea.","The electrical currents and electric fields from each electrode stimulate the nerves disposed on the modiolus of the cochlea.","Despite the enormous benefits offered by cochlear implants, one potential disadvantage of placement of the electrode assembly within the scala tympani is that it is necessary to breach the internal ducts of the cochlea, generally the scala tympani.","The breaching of the scala tympani of the cochlea adversely affects the hydrodynamic behaviour of the cochlea and is thought to prevent or at least reduce any chance of preservation of any residual hearing of the implantee.","This can be problematic for those persons who would benefit from use of a cochlear implant to improve hearing of relatively high frequencies but who have some residual hearing of relatively low frequencies.","In such a case, the implantee is forced to trade off an existing residual capacity to hear relatively low frequency sounds against the desirability of being able to have a hearing sensation of relatively high frequency sounds offered by a cochlear implant.","There have been a number of proposals put forward to provide a hybrid system whereby a cochlear implant system can be used in conjunction with residual hearing, usually assisted by the use of a hearing aid.","One such example of a proposed system is described in International Patent Application No WO 00/69512.In this application, the hybrid system utilises a hearing aid to amplify the low frequency sound enabling the user to rely on normal hearing processes to experience such sounds.","For high frequency sounds, the hybrid system utilises a relatively conventional cochlear stimulation device consisting of a short cochlear electrode array.","The short cochlear electrode array of this application is described as consisting of 4-8 electrodes and is inserted directly through the round window membrane making contact with the basal region of the cochlea.","Therefore the system as described in this application still uses a relatively obtrusive electrode array making it very difficult to preserve any residual hearing the patient may have in such areas.","As already described, the present application is also directed to a device for masking or treating tinnitus.","Tinnitus is the medical term for a condition in which sufferers report a ringing in their ears or head when there is in fact no external sound present in the sufferer's audible range.","Although some people hear a ringing noise, others report the noise as being a hissing, a chirping, or a clicking.","There are various estimates as to how many sufferers of tinnitus there are worldwide.","For example, it is suggested that some 50 million Americans suffer from tinnitus, with about 83% of them hearing a constant ringing.","Other figures suggest at least 12 million people have tinnitus to what is regarded as a distressing degree.","For some people, tinnitus is just a nuisance.","For others, it can be a quite debilitating condition.","Usually, the only relief tinnitus sufferers will experience is an occasional reduction in the loudness of the tinnitus from time to time.","The cause of tinnitus or at least its onset is unclear.","There is, however, data available that demonstrates that exposure to loud noise is a trigger for the condition.","Other suggested triggers include severe head trauma, certain medications, sinus and respiratory infections, ear infections, wax build-up and certain types of tumours.","There are, as yet, no cures for tinnitus but there are several treatments currently used to provide at least some relief.","One treatment is the use of what are commonly referred to as tinnitus maskers.","One example of a tinnitus masker is disclosed in PCT Patent Application WO 90/07251.Tinnitus maskers are essentially small battery operated devices which are worn like a hearing aid behind or in the ear, and cover (mask) the tinnitus psychoacoustically by artificial sounds which are emitted, for example, via a hearing aid speaker into the auditory canal and which reduce the disturbing tinnitus as far as possible below the threshold of perception.","The artificial sounds are often narrowband noise (for example, third octave noise) which in its spectral position and its loudness level can be adjusted via a programming device to enable the maximum position adaptation to the individual tinnitus situation.","This form of treatment is available in several forms and when properly administered, has been demonstrated to assist in somewhere between 58% and 65% of cases.","Masking is simply the addition of an outside sound that serves as a substitute or mask for the tinnitus.","Masking systems known to date are typically worn within the ear canal or positioned nearby so as to ensure provision of a masking sound to the sufferer and as a result these devices stigmatise the wearer and are worn reluctantly.","Implantable tinnitus maskers are known, such as that described in U.S. Pat.","No.","5,795,287.Such devices utilise electromechanical transducers coupled to the ossicular chain to produce the artificial masking sounds, however, these devices require a very complicated surgery to implant as the electromechanical transducer must mechanically manipulate the ossicular chain.","Also, it has been found that such mechanical coupling is not always guaranteed to be stable as pressure necroses in the area of the middle ear ossicle has been found to occur in a number of cases resulting in bone erosion.","The present invention relates to a new system for treating the symptoms of tinnitus that preferably does not require complicated surgery nor the fixation of electromechanical transducers to the ossicles.","Any discussion of documents, acts, materials, devices, articles or the like which has been included in the present specification is solely for the purpose of providing a context for the present invention.","It is not to be taken as an admission that any or all of these matters form part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed before the priority date of each claim of this application."],["<SOH> SUMMARY OF THE INVENTION <EOH>Throughout this specification the word “comprise”, or variations such as “comprises” or “comprising”, will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps.","The present invention is firstly directed to an implant that can be inserted in the cochlea but in a position external to the scala tympani.","Such an implant provides an alternative option for those persons described above who would benefit from the use of a cochlear implant to improve hearing of relatively high frequencies but who have some residual hearing of relatively low frequencies.","The present invention further preferably aims to provide a cochlear implant system that preserves the normal hydrodynamic nature of the cochlea allowing for an electrode array to be positioned to stimulate the desired neurons without causing damage to the important internal ducts of the cochlea.","The present invention is secondly directed to an implant, as described above, that can be used as a means of masking the symptoms of tinnitus.","According to a first aspect, the present application is directed to a first invention comprising an implantable tissue-stimulating device having a carrier member having at least one electrode thereon, the carrier member being adapted for intracochlear but extraluminar insertion within the cochlea of an implantee.","According to a second aspect, the present invention is directed to a second invention comprising an implantable tissue-stimulating device for use in the masking or treatment of the symptoms of tinnitus, the device having a carrier member having at least one electrode thereon, the carrier member being adapted for intracochlear but extraluminar insertion within the cochlea of an implantee.","According to a third aspect, the present invention is directed to a third invention comprising an implantable tissue-stimulating device when used in the masking or treatment of the symptoms of tinnitus, the device having a carrier member having at least one electrode thereon, the carrier member being adapted for intracochlear but extraluminar insertion within the cochlea of an implantee.","In the above aspects, the device can be a cochlear implant In one embodiment, the carrier member can be a cochlear implant carrier member.","The carrier member preferably has a body having a plurality of electrodes mounted thereon.","The electrodes can be disposed in an array on the carrier member.","The electrodes can be adapted to apply a preselected tissue stimulation.","In a preferred embodiment, the carrier member is adapted to be implanted in a crevice between the spiral ligament and the endosteum of the lateral wall of the cochlea.","This is quite different to the normal placement of the electrode array of a traditional cochlear implant in the scala tympani of the cochlea.","The placement of the device is preferably designed to avoid any breach of the internal ducts of the cochlea (eg.","scala tympani and scala vestibule) so that the normal hydrodynamic behaviour of the cochlea is not affected by any intrusive device.","This is important, as for implantees suffering tinnitus, use of the device does not lead to loss of what otherwise may be good hearing.","For implantees with at least some sensorineural hearing loss, use of the device maximises the possibility of also preserving residual hearing offered by the implantee's cochlea.","In this case, it is envisaged that use of the device will have particular benefit in those instances where the implantee has substantial residual hearing in the low frequencies but would benefit from supplemental stimulation in a relatively higher frequency range.","In this case, the implantee may benefit from use of a hearing aid that amplifies the relatively low frequencies still detectable by the implantee and a cochlear implant for detection of relatively high frequencies.","In a preferred embodiment, the carrier member has a maximum length of about 7-10 mm, a width of about 0.6 mm and a thickness no greater than about 0.2 mm and, more preferably, about 0.1 mm.","In a further embodiment, the carrier member can have an inner surface and an outer surface, the inner surface being adapted to face inwardly into the cochlea, while the outer surface faces toward the endosteum of the cochlea.","In one embodiment, the inner face can have a concavity.","In a further embodiment, the outer face can have a convexity.","In a further embodiment, the thickness of the carrier member between its inner surface and outer surface can be substantially constant for at least a majority of its length from the proximal end to the distal end.","In another embodiment, the thickness of the carrier can change, such as decrease, from the proximal end to the distal end.","In a preferred embodiment, the carrier can be relatively more resiliently flexible in a longitudinal plane and relatively less resiliently flexible in a lateral plane.","The carrier member can be relatively flexible and preferably adapted to follow the curvature of the endosteum along the basal turn.","In a preferred embodiment, a proximal end of the carrier member can be identified by a stop member.","The stop member can extend substantially at right angles to the longitudinal axis of the carrier member.","The stop member preferably has a length of between about 1.5 and 2.0 mm.","The stop member can serve as both a region for grasping the carrier member and also act to prevent insertion of the carrier member within the crevice beyond a predetermined maximum depth.","In a still further embodiment, said at least one electrode has a surface that is at least adjacent the inner surface of the carrier.","More preferably, each of the electrodes in the array has a surface that is adjacent the inner surface of the carrier member.","In a further embodiment, the surfaces of the electrodes are aligned with the inner surface of the carrier member.","In another embodiment, the surfaces of the electrodes stand proud of the inner surface of the carrier member.","It is also envisaged that the electrode surface could also be recessed into the inner surface of the carrier member.","In one embodiment, the carrier member can be formed from a biocompatible elastomeric material.","In one embodiment, the elastomeric material can be a silicone rubber.","In another embodiment, the carrier member can be formed from a biocompatible polyurethane or similar material.","The surfaces of the carrier member are preferably smooth to prevent any damage to the cochlea as the array is placed in the cochlea.","In a preferred embodiment, the electrode array can include electrically conducting wires connected to the electrodes and extending to at least said proximal end.","In one embodiment, one wire can be connected to each of said electrodes.","In another embodiment, at least two wires can be connected to each of said electrodes.","Each electrode can comprise a contact element.","The carrier member can have a longitudinal axis with each contact element arranged orthogonally to the longitudinal axis.","The contact elements can be formed from a biocompatible material.","The biocompatible material of the contact element can be platinum.","The wires may preferably be connected to the contact elements by welding, or any other suitable connecting method.","Once implanted, the electrodes of the carrier member preferably receive stimulation signals from a stimulator means.","The stimulator means is preferably electrically connected to the carrier member by way of an electrical lead.","The lead can include the one or more wires extending from each electrode of the array mounted on the carrier member.","In one embodiment, the lead can extend from the carrier member to the stimulator means or at least the housing thereof.","In one embodiment, the lead is continuous with no electrical connectors, at least external the housing of the.","stimulator means, required to connect the wires extending from the electrodes to the stimulator means.","One advantage of this arrangement is that there is no requirement for the surgeon implanting the device to make the necessary electrical connection between the wires extending from the electrodes and the stimulator means.","The stimulator means is preferably positioned within a housing that is implantable within the implantee.","The housing for the stimulator means is preferably implantable within a recess in the bone behind the ear posterior to the mastoid.","When implanted, the housing preferably contains, in addition to the stimulator means, a receiver means.","The receiver means is preferably adapted to receive signals from a controller means.","The controller means is, in use, preferably mounted external to the body of the implantee such that the signals are transmitted transcutaneously through the skin of the implantee.","Signals can preferably travel from the controller means to the receiver means and vice versa.","The receiver means can include a receiver coil adapted to receive radio frequency (RF) signals from a corresponding transmitter coil worn externally of the body.","The radio frequency signals can comprise frequency modulated (FM) signals.","While described as a receiver coil, the receiver coil can preferably transmit signals to the transmitter coil which receives the signals.","The transmitter coil is preferably held in position adjacent the implanted location of the receiver coil by way of respective attractive magnets mounted centrally in, or at some other position relative to, the coils.","The external controller can comprise a processor adapted to output one or more stimulation regimes to the stimulator.","Where the device is being used to mask or treat the symptoms of tinnitus, the external controller can comprise a processor adapted to output one or more stimulation regimes to the stimulator.","In one embodiment of this application, the stimulation regime can comprise a random continuous sub-threshold stimulation regime.","In this regime, the stimulation signals output to the electrodes of the carrier member are at a level below the threshold of hearing of the sufferer.","In another embodiment, the stimulation regime can comprise a random continuous supra-threshold stimulation, such as white noise.","In a still further embodiment, the stimulation regime can comprise a random discontinuous supra-threshold stimulation regime.","It is postulated by the present inventors that irregular stimulation may be sufficient to reduce the impact of the tinnitus condition.","Irregular stimulation also has the advantage of being relatively power-efficient and hence would result in longer battery life for the device.","In yet a further embodiment, the stimulation regime can comprise a treatment-on-demand regime.","Such a regime is postulated by the present inventors as being advantageous for those persons who only suffer irregular episodes of tinnitus.","In this embodiment, the external controller can further comprise an activation means.","The activation means can comprise a switch means on the external controller.","In this case, the sufferer or a third person could activate the processor when required.","Where the device is being used to provide a hearing sensation, the external controller preferably comprises a speech processor adapted to receive signals output by a microphone.","During use, the microphone is preferably worn on the pinna of the implanted, however, other suitable locations can be envisaged, such as the lapel of the implantee's clothing.","The speech processor encodes the sound detected by the microphone into a sequence of electrical stimuli following given algorithms, such as algorithms already developed for cochlear implant systems.","The encoded sequence is transferred to the implanted receiver/stimulator means using the transmitter and receiver coils.","The implanted receiver/stimulator means demodulates the modulated FM signal and allocates the electrical pulses to the appropriate attached electrode by an algorithm which is consistent with the chosen speech coding strategy.","In one embodiment, the processor can be adapted to receive signals from the microphone when external sounds, such as speech, are present and to output a stimulation regime, when no external sounds are present, that is adapted to mask or treat the symptoms of tinnitus.","The external controller preferably further comprises a power supply.","The power supply can comprise one or more rechargeable batteries.","The transmitter and receiver coils are used to provide power via transcutaneous induction to the implanted receiver/stimulator means and the electrode array.","While the implant system can rely on external componentry, in another embodiment, the controller means, including the microphone where present, the processor, and the power supply can also be implantable.","In this embodiment, the controller means can be contained within a hermetically sealed housing or the housing used for the stimulator means.","An implantable controller means also preferably has an activation means that provides the implantee or a third person with a means to activate a stimulation regime when required.","In one embodiment, the inactivation means can comprise a magnetic switch.","In this case, the implantable controller means would preferably incorporate a magnetic field detector which is triggered on detecting the presence of a suitable magnet held dose to the location of the implantable controller.","In another embodiment, the activation means can comprise a radio frequency switch means.","In this case, the implantable controller can include a radio frequency detector means adapted to receive a particular predetermined or programmed signal from a radio transmitter.","The radio transmitter is activated when required by the implantee or a third person.","In yet another embodiment, the activation means can comprise an infrared switch means.","In this case, the implantable controller can include a infrared detector means adapted to receive a particular pre-determined or programmed infrared signal from a infrared transmitter.","The infrared transmitter is activated when required by the implantee or a third person.","According to a fourth aspect, the present invention is directed to a fourth invention comprising a method of inserting a tissue-stimulating device as defined above into a cochlea of an implantee, the method comprising the steps of: performing a fenestration to access the crevice of the cochlea between the spiral ligament and the endosteum of the lateral wall of the cochlea; placing the device in the crevice; and closing the fenestration.","According to a fifth aspect, the present invention is directed to a fifth invention comprising a method of treating the symptoms of tinnitus comprising inserting a device as defined above into a cochlea of an implantee, the method comprising the steps of: performing a fenestration to access the crevice of the cochlea between the spiral ligament and the endosteum of the lateral wall of the cochlea; placing the device in the crevice; and closing the fenestration.","In a preferred embodiment of the fourth and fifth aspects, the device is guided very gently through the fenestration and into the crevice.","Following placement of the device, the method can include the step of covering the fenestration with soft body tissue and/or bony dust mixed with fibrin.","Once in place, the device can be used to output-one or more stimulation regimes to the cochlea to provide a hearing sensation and/or mask or treat the symptoms of tinnitus as described above."],["FIELD OF THE INVENTION The present invention relates to an auditory prosthesis adapted for intracochlear but extraluminar placement within the cochlea.","An application of the device for masking or treating tinnitus is also described.","BACKGROUND OF THE INVENTION Hearing loss, which may be due to many different causes, is generally of two types, conductive and sensorineural.","In some cases, a person may have hearing loss of both types.","Of them, conductive hearing loss occurs where the normal mechanical pathways for sound to reach the hair cells in the cochlea are impeded, for example, by damage to the ossicdes.","Conductive hearing loss may often be helped by use of conventional hearing aids, which amplify sound so that acoustic information does reach the cochlea and the hair cells.","In many people who are profoundly deaf, however, the reason for their deafness is sensorineural hearing loss.","This type of hearing loss is due to the absence of, or destruction of, the hair cells in the cochlea which transduce acoustic signals into nerve impulses.","These people are thus unable to derive suitable benefit from conventional hearing aid systems, no matter how loud the acoustic stimulus is made, because there is damage to or absence of the mechanism for nerve impulses to be generated from sound in the normal manner.","It is for this purpose that cochlear implant systems have been developed.","Such systems bypass the hair cells in the cochlea and directly deliver electrical stimulation to the auditory nerve fibres, thereby allowing the brain to perceive a hearing sensation resembling the natural hearing sensation normally delivered to the auditory nerve.","U.S. Pat.","No.","4,532,930, the contents of which are incorporated herein by reference, provides a description of one type of traditional cochlear implant system.","Typically, cochlear implant systems have consisted of essentially two components, an external component commonly referred to as a processor unit and an internal implanted component commonly referred to as a receiver/stimulator unit.","Traditionally, both of these components have cooperated together to provide the sound sensation to a user.","The external component has traditionally consisted of a microphone for detecting sounds, such as speech and environmental sounds, a speech processor that converts speech into a coded signal, a power source such as a battery, and an external transmitter coil.","The coded signal output by the sound processor is transmitted transcutaneously to the implanted receiver/stimulator unit situated within a recess of the temporal bone of the user.","This transcutaneous transmission occurs via the external transmitter coil which is positioned to communicate with an implanted receiver coil provided with the stimulator/receiver unit.","This communication serves two essential purposes, firstly to transcutaneously transmit the coded sound signal and secondly to provide power to the implanted receiver/stimulator unit Conventionally, this link has been in the form of a radio frequency (RF) link, but other such links have been proposed and implemented with varying degrees of success.","The implanted receiver/stimulator unit traditionally includes a receiver coil that receives the coded signal and power from the external processor component, and a stimulator that processes the coded signal and outputs a stimulation signal to an intracochlea electrode assembly which applies the electrical stimulation directly to the auditory nerve producing a hearing sensation corresponding to the original detected sound.","Traditionally, at least the speech processor of the external componentry has been carried on the body of the user, such as in a pocket of the user's clothing, a belt pouch or in a harness, while the microphone has been mounted on a clip mounted behind the ear or on the lapel of the user.","More recently, due in the main to improvements in technology, the physical dimensions of the sound processor have been able to be reduced allowing for the external componentry to be housed in a small unit capable of being worn behind the ear of the user.","This unit allows the microphone, power unit and the sound processor to be housed in a single unit capable of being discretely worn behind the ear, with the external transmitter coil still positioned on the side of the user's head to allow for the transmission of the coded sound signal from the sound processor and power to the implanted stimulator unit.","It is further envisaged that with continual improvements in technology all the traditional external componentry may be implanted in the user.","In such a system all of the speech processing may be performed inside the implanted stimulator unit, via an implanted microphone.","It is known in the art that the cochlea is tonotopically mapped.","In other words, the cochlea can be partitioned into regions, with each region being responsive to signals in a particular frequency range.","This property of the cochlea has been exploited by providing the electrode assembly with an array of electrodes, each electrode being arranged and constructed to deliver a stimulating signal within a preselected frequency range, to the appropriate region within the scala tympani of the cochlea.","The electrical currents and electric fields from each electrode stimulate the nerves disposed on the modiolus of the cochlea.","Despite the enormous benefits offered by cochlear implants, one potential disadvantage of placement of the electrode assembly within the scala tympani is that it is necessary to breach the internal ducts of the cochlea, generally the scala tympani.","The breaching of the scala tympani of the cochlea adversely affects the hydrodynamic behaviour of the cochlea and is thought to prevent or at least reduce any chance of preservation of any residual hearing of the implantee.","This can be problematic for those persons who would benefit from use of a cochlear implant to improve hearing of relatively high frequencies but who have some residual hearing of relatively low frequencies.","In such a case, the implantee is forced to trade off an existing residual capacity to hear relatively low frequency sounds against the desirability of being able to have a hearing sensation of relatively high frequency sounds offered by a cochlear implant.","There have been a number of proposals put forward to provide a hybrid system whereby a cochlear implant system can be used in conjunction with residual hearing, usually assisted by the use of a hearing aid.","One such example of a proposed system is described in International Patent Application No WO 00/69512.In this application, the hybrid system utilises a hearing aid to amplify the low frequency sound enabling the user to rely on normal hearing processes to experience such sounds.","For high frequency sounds, the hybrid system utilises a relatively conventional cochlear stimulation device consisting of a short cochlear electrode array.","The short cochlear electrode array of this application is described as consisting of 4-8 electrodes and is inserted directly through the round window membrane making contact with the basal region of the cochlea.","Therefore the system as described in this application still uses a relatively obtrusive electrode array making it very difficult to preserve any residual hearing the patient may have in such areas.","As already described, the present application is also directed to a device for masking or treating tinnitus.","Tinnitus is the medical term for a condition in which sufferers report a ringing in their ears or head when there is in fact no external sound present in the sufferer's audible range.","Although some people hear a ringing noise, others report the noise as being a hissing, a chirping, or a clicking.","There are various estimates as to how many sufferers of tinnitus there are worldwide.","For example, it is suggested that some 50 million Americans suffer from tinnitus, with about 83% of them hearing a constant ringing.","Other figures suggest at least 12 million people have tinnitus to what is regarded as a distressing degree.","For some people, tinnitus is just a nuisance.","For others, it can be a quite debilitating condition.","Usually, the only relief tinnitus sufferers will experience is an occasional reduction in the loudness of the tinnitus from time to time.","The cause of tinnitus or at least its onset is unclear.","There is, however, data available that demonstrates that exposure to loud noise is a trigger for the condition.","Other suggested triggers include severe head trauma, certain medications, sinus and respiratory infections, ear infections, wax build-up and certain types of tumours.","There are, as yet, no cures for tinnitus but there are several treatments currently used to provide at least some relief.","One treatment is the use of what are commonly referred to as tinnitus maskers.","One example of a tinnitus masker is disclosed in PCT Patent Application WO 90/07251.Tinnitus maskers are essentially small battery operated devices which are worn like a hearing aid behind or in the ear, and cover (mask) the tinnitus psychoacoustically by artificial sounds which are emitted, for example, via a hearing aid speaker into the auditory canal and which reduce the disturbing tinnitus as far as possible below the threshold of perception.","The artificial sounds are often narrowband noise (for example, third octave noise) which in its spectral position and its loudness level can be adjusted via a programming device to enable the maximum position adaptation to the individual tinnitus situation.","This form of treatment is available in several forms and when properly administered, has been demonstrated to assist in somewhere between 58% and 65% of cases.","Masking is simply the addition of an outside sound that serves as a substitute or mask for the tinnitus.","Masking systems known to date are typically worn within the ear canal or positioned nearby so as to ensure provision of a masking sound to the sufferer and as a result these devices stigmatise the wearer and are worn reluctantly.","Implantable tinnitus maskers are known, such as that described in U.S. Pat.","No.","5,795,287.Such devices utilise electromechanical transducers coupled to the ossicular chain to produce the artificial masking sounds, however, these devices require a very complicated surgery to implant as the electromechanical transducer must mechanically manipulate the ossicular chain.","Also, it has been found that such mechanical coupling is not always guaranteed to be stable as pressure necroses in the area of the middle ear ossicle has been found to occur in a number of cases resulting in bone erosion.","The present invention relates to a new system for treating the symptoms of tinnitus that preferably does not require complicated surgery nor the fixation of electromechanical transducers to the ossicles.","Any discussion of documents, acts, materials, devices, articles or the like which has been included in the present specification is solely for the purpose of providing a context for the present invention.","It is not to be taken as an admission that any or all of these matters form part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed before the priority date of each claim of this application.","SUMMARY OF THE INVENTION Throughout this specification the word “comprise”, or variations such as “comprises” or “comprising”, will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps.","The present invention is firstly directed to an implant that can be inserted in the cochlea but in a position external to the scala tympani.","Such an implant provides an alternative option for those persons described above who would benefit from the use of a cochlear implant to improve hearing of relatively high frequencies but who have some residual hearing of relatively low frequencies.","The present invention further preferably aims to provide a cochlear implant system that preserves the normal hydrodynamic nature of the cochlea allowing for an electrode array to be positioned to stimulate the desired neurons without causing damage to the important internal ducts of the cochlea.","The present invention is secondly directed to an implant, as described above, that can be used as a means of masking the symptoms of tinnitus.","According to a first aspect, the present application is directed to a first invention comprising an implantable tissue-stimulating device having a carrier member having at least one electrode thereon, the carrier member being adapted for intracochlear but extraluminar insertion within the cochlea of an implantee.","According to a second aspect, the present invention is directed to a second invention comprising an implantable tissue-stimulating device for use in the masking or treatment of the symptoms of tinnitus, the device having a carrier member having at least one electrode thereon, the carrier member being adapted for intracochlear but extraluminar insertion within the cochlea of an implantee.","According to a third aspect, the present invention is directed to a third invention comprising an implantable tissue-stimulating device when used in the masking or treatment of the symptoms of tinnitus, the device having a carrier member having at least one electrode thereon, the carrier member being adapted for intracochlear but extraluminar insertion within the cochlea of an implantee.","In the above aspects, the device can be a cochlear implant In one embodiment, the carrier member can be a cochlear implant carrier member.","The carrier member preferably has a body having a plurality of electrodes mounted thereon.","The electrodes can be disposed in an array on the carrier member.","The electrodes can be adapted to apply a preselected tissue stimulation.","In a preferred embodiment, the carrier member is adapted to be implanted in a crevice between the spiral ligament and the endosteum of the lateral wall of the cochlea.","This is quite different to the normal placement of the electrode array of a traditional cochlear implant in the scala tympani of the cochlea.","The placement of the device is preferably designed to avoid any breach of the internal ducts of the cochlea (eg.","scala tympani and scala vestibule) so that the normal hydrodynamic behaviour of the cochlea is not affected by any intrusive device.","This is important, as for implantees suffering tinnitus, use of the device does not lead to loss of what otherwise may be good hearing.","For implantees with at least some sensorineural hearing loss, use of the device maximises the possibility of also preserving residual hearing offered by the implantee's cochlea.","In this case, it is envisaged that use of the device will have particular benefit in those instances where the implantee has substantial residual hearing in the low frequencies but would benefit from supplemental stimulation in a relatively higher frequency range.","In this case, the implantee may benefit from use of a hearing aid that amplifies the relatively low frequencies still detectable by the implantee and a cochlear implant for detection of relatively high frequencies.","In a preferred embodiment, the carrier member has a maximum length of about 7-10 mm, a width of about 0.6 mm and a thickness no greater than about 0.2 mm and, more preferably, about 0.1 mm.","In a further embodiment, the carrier member can have an inner surface and an outer surface, the inner surface being adapted to face inwardly into the cochlea, while the outer surface faces toward the endosteum of the cochlea.","In one embodiment, the inner face can have a concavity.","In a further embodiment, the outer face can have a convexity.","In a further embodiment, the thickness of the carrier member between its inner surface and outer surface can be substantially constant for at least a majority of its length from the proximal end to the distal end.","In another embodiment, the thickness of the carrier can change, such as decrease, from the proximal end to the distal end.","In a preferred embodiment, the carrier can be relatively more resiliently flexible in a longitudinal plane and relatively less resiliently flexible in a lateral plane.","The carrier member can be relatively flexible and preferably adapted to follow the curvature of the endosteum along the basal turn.","In a preferred embodiment, a proximal end of the carrier member can be identified by a stop member.","The stop member can extend substantially at right angles to the longitudinal axis of the carrier member.","The stop member preferably has a length of between about 1.5 and 2.0 mm.","The stop member can serve as both a region for grasping the carrier member and also act to prevent insertion of the carrier member within the crevice beyond a predetermined maximum depth.","In a still further embodiment, said at least one electrode has a surface that is at least adjacent the inner surface of the carrier.","More preferably, each of the electrodes in the array has a surface that is adjacent the inner surface of the carrier member.","In a further embodiment, the surfaces of the electrodes are aligned with the inner surface of the carrier member.","In another embodiment, the surfaces of the electrodes stand proud of the inner surface of the carrier member.","It is also envisaged that the electrode surface could also be recessed into the inner surface of the carrier member.","In one embodiment, the carrier member can be formed from a biocompatible elastomeric material.","In one embodiment, the elastomeric material can be a silicone rubber.","In another embodiment, the carrier member can be formed from a biocompatible polyurethane or similar material.","The surfaces of the carrier member are preferably smooth to prevent any damage to the cochlea as the array is placed in the cochlea.","In a preferred embodiment, the electrode array can include electrically conducting wires connected to the electrodes and extending to at least said proximal end.","In one embodiment, one wire can be connected to each of said electrodes.","In another embodiment, at least two wires can be connected to each of said electrodes.","Each electrode can comprise a contact element.","The carrier member can have a longitudinal axis with each contact element arranged orthogonally to the longitudinal axis.","The contact elements can be formed from a biocompatible material.","The biocompatible material of the contact element can be platinum.","The wires may preferably be connected to the contact elements by welding, or any other suitable connecting method.","Once implanted, the electrodes of the carrier member preferably receive stimulation signals from a stimulator means.","The stimulator means is preferably electrically connected to the carrier member by way of an electrical lead.","The lead can include the one or more wires extending from each electrode of the array mounted on the carrier member.","In one embodiment, the lead can extend from the carrier member to the stimulator means or at least the housing thereof.","In one embodiment, the lead is continuous with no electrical connectors, at least external the housing of the.","stimulator means, required to connect the wires extending from the electrodes to the stimulator means.","One advantage of this arrangement is that there is no requirement for the surgeon implanting the device to make the necessary electrical connection between the wires extending from the electrodes and the stimulator means.","The stimulator means is preferably positioned within a housing that is implantable within the implantee.","The housing for the stimulator means is preferably implantable within a recess in the bone behind the ear posterior to the mastoid.","When implanted, the housing preferably contains, in addition to the stimulator means, a receiver means.","The receiver means is preferably adapted to receive signals from a controller means.","The controller means is, in use, preferably mounted external to the body of the implantee such that the signals are transmitted transcutaneously through the skin of the implantee.","Signals can preferably travel from the controller means to the receiver means and vice versa.","The receiver means can include a receiver coil adapted to receive radio frequency (RF) signals from a corresponding transmitter coil worn externally of the body.","The radio frequency signals can comprise frequency modulated (FM) signals.","While described as a receiver coil, the receiver coil can preferably transmit signals to the transmitter coil which receives the signals.","The transmitter coil is preferably held in position adjacent the implanted location of the receiver coil by way of respective attractive magnets mounted centrally in, or at some other position relative to, the coils.","The external controller can comprise a processor adapted to output one or more stimulation regimes to the stimulator.","Where the device is being used to mask or treat the symptoms of tinnitus, the external controller can comprise a processor adapted to output one or more stimulation regimes to the stimulator.","In one embodiment of this application, the stimulation regime can comprise a random continuous sub-threshold stimulation regime.","In this regime, the stimulation signals output to the electrodes of the carrier member are at a level below the threshold of hearing of the sufferer.","In another embodiment, the stimulation regime can comprise a random continuous supra-threshold stimulation, such as white noise.","In a still further embodiment, the stimulation regime can comprise a random discontinuous supra-threshold stimulation regime.","It is postulated by the present inventors that irregular stimulation may be sufficient to reduce the impact of the tinnitus condition.","Irregular stimulation also has the advantage of being relatively power-efficient and hence would result in longer battery life for the device.","In yet a further embodiment, the stimulation regime can comprise a treatment-on-demand regime.","Such a regime is postulated by the present inventors as being advantageous for those persons who only suffer irregular episodes of tinnitus.","In this embodiment, the external controller can further comprise an activation means.","The activation means can comprise a switch means on the external controller.","In this case, the sufferer or a third person could activate the processor when required.","Where the device is being used to provide a hearing sensation, the external controller preferably comprises a speech processor adapted to receive signals output by a microphone.","During use, the microphone is preferably worn on the pinna of the implanted, however, other suitable locations can be envisaged, such as the lapel of the implantee's clothing.","The speech processor encodes the sound detected by the microphone into a sequence of electrical stimuli following given algorithms, such as algorithms already developed for cochlear implant systems.","The encoded sequence is transferred to the implanted receiver/stimulator means using the transmitter and receiver coils.","The implanted receiver/stimulator means demodulates the modulated FM signal and allocates the electrical pulses to the appropriate attached electrode by an algorithm which is consistent with the chosen speech coding strategy.","In one embodiment, the processor can be adapted to receive signals from the microphone when external sounds, such as speech, are present and to output a stimulation regime, when no external sounds are present, that is adapted to mask or treat the symptoms of tinnitus.","The external controller preferably further comprises a power supply.","The power supply can comprise one or more rechargeable batteries.","The transmitter and receiver coils are used to provide power via transcutaneous induction to the implanted receiver/stimulator means and the electrode array.","While the implant system can rely on external componentry, in another embodiment, the controller means, including the microphone where present, the processor, and the power supply can also be implantable.","In this embodiment, the controller means can be contained within a hermetically sealed housing or the housing used for the stimulator means.","An implantable controller means also preferably has an activation means that provides the implantee or a third person with a means to activate a stimulation regime when required.","In one embodiment, the inactivation means can comprise a magnetic switch.","In this case, the implantable controller means would preferably incorporate a magnetic field detector which is triggered on detecting the presence of a suitable magnet held dose to the location of the implantable controller.","In another embodiment, the activation means can comprise a radio frequency switch means.","In this case, the implantable controller can include a radio frequency detector means adapted to receive a particular predetermined or programmed signal from a radio transmitter.","The radio transmitter is activated when required by the implantee or a third person.","In yet another embodiment, the activation means can comprise an infrared switch means.","In this case, the implantable controller can include a infrared detector means adapted to receive a particular pre-determined or programmed infrared signal from a infrared transmitter.","The infrared transmitter is activated when required by the implantee or a third person.","According to a fourth aspect, the present invention is directed to a fourth invention comprising a method of inserting a tissue-stimulating device as defined above into a cochlea of an implantee, the method comprising the steps of: performing a fenestration to access the crevice of the cochlea between the spiral ligament and the endosteum of the lateral wall of the cochlea; placing the device in the crevice; and closing the fenestration.","According to a fifth aspect, the present invention is directed to a fifth invention comprising a method of treating the symptoms of tinnitus comprising inserting a device as defined above into a cochlea of an implantee, the method comprising the steps of: performing a fenestration to access the crevice of the cochlea between the spiral ligament and the endosteum of the lateral wall of the cochlea; placing the device in the crevice; and closing the fenestration.","In a preferred embodiment of the fourth and fifth aspects, the device is guided very gently through the fenestration and into the crevice.","Following placement of the device, the method can include the step of covering the fenestration with soft body tissue and/or bony dust mixed with fibrin.","Once in place, the device can be used to output-one or more stimulation regimes to the cochlea to provide a hearing sensation and/or mask or treat the symptoms of tinnitus as described above.","BRIEF DESCRIPTION OF THE DRAWINGS By way of example only, a preferred mode of carrying out the invention is described with reference to the accompanying drawings, in which: FIG.","1 is a perspective view of one embodiment of a carrier member of an implantable electrode array according to the present invention;: FIG.","2 is a cross-sectional view through a human cochlea depicting the desired position for placement of the electrode array according to the present invention; and FIG.","3 is a pictorial representation of one embodiment of a system that can utilise the carrier member of FIG.","1 whether it be for providing a hearing sensation to an implantee and/or to mask or treat the symptoms of tinnitus.","PREFERRED MODE OF CARRYING OUT THE INVENTION One embodiment of an electrode assembly for use in the present invention according to the present invention is depicted generally as 10 in the drawings.","The assembly 10 includes an elongate electrode carrier member 11.For the purposes of clarity, the plurality of electrodes that are mounted on the carrier member 11 are not depicted in the drawings.","While not depicted, the electrodes can be disposed in a linear array on the carrier member 11 and be adapted to apply a preselected tissue stimulation to the cochlea.","The depicted carrier member 11 is preformed from a resiliently flexible biocompatible silicone and extends from a distal end 12 to a stop member 13.The carrier member 11 is adapted for intracochlear but extraluminar insertion within the cochlea of an implantee.","In use, the carrier member 11 is adapted to be implanted in the crevice 21 (see FIG.","2) between the spiral ligament 22 and the endosteum 23 of the lateral wall of the cochlea 20.This is a quite different location to the normal placement of the cochlear implant electrode array in the scala tympani 24 of the cochlea 20.The placement of the carrier member 11 is designed to avoid any breach of the internal ducts of the cochlea 20 (eg.","scala tympani 24 and scala vestibuli 25) so that the normal hydrodynamic behaviour of the cochlea 20 is not affected by any intrusive device.","By preserving the normal hydrodynamic behaviour of the cochlea 20, use of the carrier member 11 maximises the possibility of also preserving any hearing of the implantee that is offered by the cochlea 20.In the depicted embodiment, the carrier member 11 has a maximum length of about 8-10 mm, a width of about 0.6 mm and a thickness no greater than about 0.1 mm.","The thickness of the carrier member between its inner surface and outer surface is substantially constant for at least a majority of its length from the distal end 12 to the stop member 13, except in the region adjacent the end 12 where the thickness gradually tapers towards end 12.The depicted carrier 11 is more resiliently flexible in a longitudinal plane and relatively less resiliently flexible in a lateral plane.","The carrier member 11 is adapted to follow the curvature of the endosteum along the basal turn of the cochlea 20.As depicted, the stop member 13 extends substantially at right angles to the longitudinal axis of the carrier member 11.The depicted stop member 13 has a length of between about 1.5 and 2.0 mm.","The stop member 13 serves as both a region for grasping the carrier member 11 and also acts to prevent insertion of the carrier member 11 within the crevice 21 beyond a predetermined maximum depth (eg.","8-10 mm).","While not depicted, the electrode assembly 10 includes electrically conducting wires connected to the electrodes and extending at least beyond the stop member 13.In the depicted embodiment, one wire can be connected to each of said electrodes.","In use, a surgeon would perform a fenestration to access the crevice 21 between the spiral ligament 22 and the endosteum 23 of the lateral wall of the cochlea 20.The carrier member 11 would then be inserted into the crevice 21 using an insertion tool that grips the stop member 13 and guides the carrier member 11 very gently into the crevice 21.Once positioned, the surgeon could close the fenestration by covering it with soft body tissue and/or bony dust mixed with fibrin.","Once implanted, the electrodes of the carrier member 11 receive stimulation signals from an implantable stimulator 30.The stimulator 30 is positioned within a housing 31 that is implantable within the implantee.","The housing 31 for the stimulator 30 is implantable within a recess in the bone behind the ear posterior to the mastoid.","When implanted, the housing 31 contains, in addition to the stimulator 30, a receiver.","The receiver is adapted to receive signals from a controller 32.The controller 32 is, in use, typically mounted external to the body 33 of the implantee such that the signals are transmitted transcutaneously through the skin of the implantee.","The controller 32 can be worn on the body of the user or can be adapted to be worn behind the ear of the user in much the same way as conventional hearing aid devices.","The signals travel from the controller 32 to the receiver and vice versa by use of a receiver coil 34 adapted to receive radio frequency (RF) signals from a corresponding transmitter coil 35 worn externally of the body 33.The radio frequency signals can comprise frequency modulated (FM) signals.","While described as a receiver coil, the receiver coil 34 can also transmit signals to the transmitter coil 35 which receives the signals.","The transmitter coil 35 is held in position adjacent the implanted location of the receiver coil 34 by way of respective attractive magnets mounted centrally in, or at some other position relative to, the coils.","The external controller 32 comprises a processor adapted to output one or more stimulation regimes to the stimulator 31.It is also envisaged that the stimulator 31 may have the capability to store one or more stimulation regimes and upon request deliver the regime to the user via the electrodes.","In one use of the system, the stimulation regimes of the stimulator 31 are designed to mask or treat the symptoms of tinnitus.","In one embodiment, the stimulation regime can comprise a random continuous sub-threshold stimulation regime.","In this regime, the stimulation signals output to the electrodes of the carrier member 11 are at level below the threshold of hearing of the sufferer.","In another embodiment, the stimulation regime can comprise a random continuous supra-threshold stimulation, such as white noise.","In a still further embodiment, the stimulation regime can comprise a random discontinuous supra-threshold stimulation regime.","In yet a further embodiment, the stimulation regime can comprise a treatment-on-demand regime.","In this embodiment, the external controller 32 can further comprise an activation means 36.The depicted activation means 36 comprises a switch on the external controller 32.In this case, the sufferer or a third person could activate the processor when required.","In addition to the above, the processor can be adapted to receive signals output by a microphone (not depicted).","In this case and during use, the microphone can be worn on the pinna of the implantee, however, other-suitable locations can be envisaged, such as a lapel of the implantee's clothing.","In this case, the processor can encode the sound detected by the microphone into a sequence of electrical stimuli following given algorithms, such as algorithms already developed for cochlear implant systems.","The encoded sequence is then transferred to the implanted stimulator 30 using the transmitter and receiver coils.","The implanted stimulator 30 demodulates the FM signals and allocates the electrical pulses to the appropriate attached electrode by an algorithm which is consistent with the chosen speech coding strategy.","The housing of the external controller 32 further houses a power supply.","In the depicted embodiment, the power supply comprise one or more rechargeable batteries.","The transmitter and receiver coils are used to provide power via transcutaneous induction to the implanted stimulator 30 and the electrode array 10.While the implant system can rely on external componentry, in another embodiment, the controller, including the microphone where present, the processor, and the power supply can also be implantable.","In this embodiment, which is not depicted, the controller can be contained within a hermetically sealed housing or the housing used for the stimulator.","An implantable controller means would typically have an activation means that provides the implantee or a third person with a means to activate a stimulation regime when required.","In one embodiment, the inactivation means can comprise a magnetic switch.","In this case, the implantable controller would preferably incorporate a magnetic field detector which is triggered on detecting the presence of a suitable magnet held close to the location of the implantable controller.","In another embodiment, the activation means can comprise a radio frequency switch means.","In this case, the implantable controller can include a radio frequency detector means adapted to receive a particular predetermined or programmed signal from a radio transmitter.","The radio transmitter is activated when required by the implantee or a third person.","In yet another embodiment, the activation means can comprise an infrared switch means.","In this case, the implantable controller can include a infrared detector means adapted to receive a particular predetermined or programmed infrared signal from a infrared transmitter.","The infrared transmitter is activated when required by the implantee or a third person.","The present invention provides an implantable device that can be used to provide a hearing sensation to an implantee with sensorineural hearing loss and or mask or treat the symptoms of tinnitus while preserving the hearing of the implantee's cochlea into which the stimulating electrode array has been inserted.","It will be appreciated by persons skilled in the art that numerous variations and/or modifications may be made to the invention as shown in the specific embodiments without departing from the spirit or scope of the invention as broadly described.","The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive."]],"string":"[\n [\n \"Endosteal electrode\",\n \"An implantable tissue-stimulating device comprising an elongate electrode carrier member (11) having a plurality of electrodes thereon.\",\n \"The electrodes are preferably disposed in a linear array on the carrier member (11) and are adapted to apply a preselected tissue stimulation to the cochlea.\",\n \"The carrier member (11) is preformed from a resiliently flexible biocompatible silicone and extends from a distal end (12) to a stop member (13).\",\n \"The carrier member (11) is adapted for intracochlear but extraluminar insertion within the cochlea of an implantee.\",\n \"In particular, the carrier member (11) is adapted to be implanted in the crevice (21) between the spiral ligament (22) and the endosteum (23) of the lateral wall of the cochlea (20).\",\n \"This is a quite different location to the normal placement of the cochlear implant electrode array in the scala tympani (24) of the cochlea (20).\",\n \"The placement of the carrier member (11) is designed to avoid any breach of the internal ducts of the cochlea (20), such as the scala tympani (24) and scala vestibuli (25) so that the normal hydrodynamic behaviour of the cochlea (20) is not affected by any intrusive device.\",\n \"By preservng the normal hydrodynamic behaviour of the cochlea (20), use of the carrier member (11) maximises the possibility of also preserving any hearing of the implantee that is offered by the cochlea (20).\",\n \"Use of the device in a system for masking or treating the symptoms of tinnitus is also described.\"\n ],\n [\n \"1.An implantable tissue-stimulating device having a carrier member having at least one electrode thereon, the carrier member being adapted for intracochlear but extraluminar insertion within the cochlea of an implantee.\",\n \"2.An implantable tissue-stimulating device for use in the masking or treatment of the symptoms of tinnitus, the device having a carrier member having at least one electrode thereon, the carrier member being adapted for intracochlear but extraluminar insertion within the cochlea of an implantee.\",\n \"3.An implantable tissue-stimulating device when used in the masking or treatment of the symptoms of tinnitus, the device having a carrier member having at least one electrode thereon, the device being adapted for intracochlear but extraluminar insertion within the cochlea of an implantee.\",\n \"4.An implantable tissue-stimulating device of claim 1 wherein the carrier member has a body having a plurality of electrodes mounted thereon.\",\n \"5.An implantable tissue-stimulating device of claim 4 wherein the electrodes are disposed in an array on the carrier member and are adapted to apply a preselected tissue stimulation.\",\n \"6.An implantable tissue-stimulating device of claim 1 wherein the carrier member is implantable in a crevice between the spiral ligament and the endosteum of the lateral wall of the cochlea.\",\n \"7.An implantable tissue-stimulating device of claim 6 wherein the carrier member has a maximum length of about 7 mm-10 mm, a width of about 0.6 mm and a thickness no greater than about 0.2 mm and, more preferably, about 0.1 mm.\",\n \"8.An implantable tissue-stimulating device of claim 6 wherein the carrier member has an inner surface and an outer surface, the inner surface being adapted on implantation to face inwardly into the cochlea.\",\n \"9.An implantable tissue-stimulating device of claim 8 wherein the inner face has a concave surface and the outer face has a convex surface.\",\n \"10.An implantable tissue-stimulating device of claim 1 wherein a stop member extends outwardly from the carrier member substantially at right angles to the longitudinal axis of the carrier member.\",\n \"11.An implantable tissue-stimulating device of claim 10 wherein the stop member has a length of between about 1.5 and 2.0 mm.\",\n \"12.An implantable tissue-stimulating device of claim 1 wherein the electrodes of the carrier member receive stimulation signals from an implantable stimulator means.\",\n \"13.An implantable tissue-stimulating device of claim 12 wherein the stimulator means is housed within a housing that also houses a receiver means that is adapted to receive signals from a controller means.\",\n \"14.An implantable tissue-stimulating device of claim 13 wherein the controller means comprises a processor means adapted to output one or more stimulation regimes to the stimulator.\",\n \"15.An implantable tissue-stimulating device of claim 14 wherein, where the device is being used to mask or treat the symptoms of tinnitus, the stimulation regime is selected from the group comprising a random continuous sub-threshold stimulation regime, a random continuous supra-threshold stimulation regime, and a random discontinuous supra-threshold stimulation regime.\",\n \"16.An implantable tissue-stimulating device of claim 14 wherein, where the device is being used to mask or treat the symptoms of tinnitus, the stimulation regime comprises a treatment-on-demand regime.\",\n \"17.An implantable tissue-stimulating device of claim 16 wherein the controller means further has an activation means that allows the implantee or a third person to activate the processor means when required.\",\n \"18.An implantable tissue-stimulating device of claim 14 wherein, where the device is being used to provide a hearing sensation, the controller means comprises a speech processor adapted to receive signals output by a microphone and encode the sound detected by the microphone into a sequence of electrical stimuli following given algorithms.\",\n \"19.An implantable tissue-stimulating device of claim 18 wherein the speech processor is adapted to receive signals from the microphone when external sounds, such as speech, are present and to output a stimulation regime, when no external sounds are present, that is adapted to mask or treat the symptoms of tinnitus.\",\n \"20.An implantable tissue-stimulating device of claim 14 wherein the controller means is implantable and contained within a hermetically sealed housing or a housing used for the stimulator means.\",\n \"21.An implantable tissue-stimulating device of claim 20 wherein the controller means has an activation means that provides the implantee or a third person with a means to activate a stimulation regime when required.\",\n \"22.A method of inserting a tissue-stimulating device as defined in claim 1 into a cochlea of an implantee, the method comprising the steps of: performing a fenestration to access the crevice of the cochlea between the spiral ligament and the endosteum of the lateral wall of the cochlea; placing the device in the crevice; and sealing the fenestration.\",\n \"23.A method of treating the symptoms of tinnitus comprising inserting a device as defined in claim 1 into a cochlea of an implantee, the method comprising the steps of: performing a fenestration to access the crevice of the cochlea between the spiral ligament and the endosteum of the lateral wall of the cochlea; placing the device in the crevice; and sealing the fenestration.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>Hearing loss, which may be due to many different causes, is generally of two types, conductive and sensorineural.\",\n \"In some cases, a person may have hearing loss of both types.\",\n \"Of them, conductive hearing loss occurs where the normal mechanical pathways for sound to reach the hair cells in the cochlea are impeded, for example, by damage to the ossicdes.\",\n \"Conductive hearing loss may often be helped by use of conventional hearing aids, which amplify sound so that acoustic information does reach the cochlea and the hair cells.\",\n \"In many people who are profoundly deaf, however, the reason for their deafness is sensorineural hearing loss.\",\n \"This type of hearing loss is due to the absence of, or destruction of, the hair cells in the cochlea which transduce acoustic signals into nerve impulses.\",\n \"These people are thus unable to derive suitable benefit from conventional hearing aid systems, no matter how loud the acoustic stimulus is made, because there is damage to or absence of the mechanism for nerve impulses to be generated from sound in the normal manner.\",\n \"It is for this purpose that cochlear implant systems have been developed.\",\n \"Such systems bypass the hair cells in the cochlea and directly deliver electrical stimulation to the auditory nerve fibres, thereby allowing the brain to perceive a hearing sensation resembling the natural hearing sensation normally delivered to the auditory nerve.\",\n \"U.S. Pat.\",\n \"No.\",\n \"4,532,930, the contents of which are incorporated herein by reference, provides a description of one type of traditional cochlear implant system.\",\n \"Typically, cochlear implant systems have consisted of essentially two components, an external component commonly referred to as a processor unit and an internal implanted component commonly referred to as a receiver/stimulator unit.\",\n \"Traditionally, both of these components have cooperated together to provide the sound sensation to a user.\",\n \"The external component has traditionally consisted of a microphone for detecting sounds, such as speech and environmental sounds, a speech processor that converts speech into a coded signal, a power source such as a battery, and an external transmitter coil.\",\n \"The coded signal output by the sound processor is transmitted transcutaneously to the implanted receiver/stimulator unit situated within a recess of the temporal bone of the user.\",\n \"This transcutaneous transmission occurs via the external transmitter coil which is positioned to communicate with an implanted receiver coil provided with the stimulator/receiver unit.\",\n \"This communication serves two essential purposes, firstly to transcutaneously transmit the coded sound signal and secondly to provide power to the implanted receiver/stimulator unit Conventionally, this link has been in the form of a radio frequency (RF) link, but other such links have been proposed and implemented with varying degrees of success.\",\n \"The implanted receiver/stimulator unit traditionally includes a receiver coil that receives the coded signal and power from the external processor component, and a stimulator that processes the coded signal and outputs a stimulation signal to an intracochlea electrode assembly which applies the electrical stimulation directly to the auditory nerve producing a hearing sensation corresponding to the original detected sound.\",\n \"Traditionally, at least the speech processor of the external componentry has been carried on the body of the user, such as in a pocket of the user's clothing, a belt pouch or in a harness, while the microphone has been mounted on a clip mounted behind the ear or on the lapel of the user.\",\n \"More recently, due in the main to improvements in technology, the physical dimensions of the sound processor have been able to be reduced allowing for the external componentry to be housed in a small unit capable of being worn behind the ear of the user.\",\n \"This unit allows the microphone, power unit and the sound processor to be housed in a single unit capable of being discretely worn behind the ear, with the external transmitter coil still positioned on the side of the user's head to allow for the transmission of the coded sound signal from the sound processor and power to the implanted stimulator unit.\",\n \"It is further envisaged that with continual improvements in technology all the traditional external componentry may be implanted in the user.\",\n \"In such a system all of the speech processing may be performed inside the implanted stimulator unit, via an implanted microphone.\",\n \"It is known in the art that the cochlea is tonotopically mapped.\",\n \"In other words, the cochlea can be partitioned into regions, with each region being responsive to signals in a particular frequency range.\",\n \"This property of the cochlea has been exploited by providing the electrode assembly with an array of electrodes, each electrode being arranged and constructed to deliver a stimulating signal within a preselected frequency range, to the appropriate region within the scala tympani of the cochlea.\",\n \"The electrical currents and electric fields from each electrode stimulate the nerves disposed on the modiolus of the cochlea.\",\n \"Despite the enormous benefits offered by cochlear implants, one potential disadvantage of placement of the electrode assembly within the scala tympani is that it is necessary to breach the internal ducts of the cochlea, generally the scala tympani.\",\n \"The breaching of the scala tympani of the cochlea adversely affects the hydrodynamic behaviour of the cochlea and is thought to prevent or at least reduce any chance of preservation of any residual hearing of the implantee.\",\n \"This can be problematic for those persons who would benefit from use of a cochlear implant to improve hearing of relatively high frequencies but who have some residual hearing of relatively low frequencies.\",\n \"In such a case, the implantee is forced to trade off an existing residual capacity to hear relatively low frequency sounds against the desirability of being able to have a hearing sensation of relatively high frequency sounds offered by a cochlear implant.\",\n \"There have been a number of proposals put forward to provide a hybrid system whereby a cochlear implant system can be used in conjunction with residual hearing, usually assisted by the use of a hearing aid.\",\n \"One such example of a proposed system is described in International Patent Application No WO 00/69512.In this application, the hybrid system utilises a hearing aid to amplify the low frequency sound enabling the user to rely on normal hearing processes to experience such sounds.\",\n \"For high frequency sounds, the hybrid system utilises a relatively conventional cochlear stimulation device consisting of a short cochlear electrode array.\",\n \"The short cochlear electrode array of this application is described as consisting of 4-8 electrodes and is inserted directly through the round window membrane making contact with the basal region of the cochlea.\",\n \"Therefore the system as described in this application still uses a relatively obtrusive electrode array making it very difficult to preserve any residual hearing the patient may have in such areas.\",\n \"As already described, the present application is also directed to a device for masking or treating tinnitus.\",\n \"Tinnitus is the medical term for a condition in which sufferers report a ringing in their ears or head when there is in fact no external sound present in the sufferer's audible range.\",\n \"Although some people hear a ringing noise, others report the noise as being a hissing, a chirping, or a clicking.\",\n \"There are various estimates as to how many sufferers of tinnitus there are worldwide.\",\n \"For example, it is suggested that some 50 million Americans suffer from tinnitus, with about 83% of them hearing a constant ringing.\",\n \"Other figures suggest at least 12 million people have tinnitus to what is regarded as a distressing degree.\",\n \"For some people, tinnitus is just a nuisance.\",\n \"For others, it can be a quite debilitating condition.\",\n \"Usually, the only relief tinnitus sufferers will experience is an occasional reduction in the loudness of the tinnitus from time to time.\",\n \"The cause of tinnitus or at least its onset is unclear.\",\n \"There is, however, data available that demonstrates that exposure to loud noise is a trigger for the condition.\",\n \"Other suggested triggers include severe head trauma, certain medications, sinus and respiratory infections, ear infections, wax build-up and certain types of tumours.\",\n \"There are, as yet, no cures for tinnitus but there are several treatments currently used to provide at least some relief.\",\n \"One treatment is the use of what are commonly referred to as tinnitus maskers.\",\n \"One example of a tinnitus masker is disclosed in PCT Patent Application WO 90/07251.Tinnitus maskers are essentially small battery operated devices which are worn like a hearing aid behind or in the ear, and cover (mask) the tinnitus psychoacoustically by artificial sounds which are emitted, for example, via a hearing aid speaker into the auditory canal and which reduce the disturbing tinnitus as far as possible below the threshold of perception.\",\n \"The artificial sounds are often narrowband noise (for example, third octave noise) which in its spectral position and its loudness level can be adjusted via a programming device to enable the maximum position adaptation to the individual tinnitus situation.\",\n \"This form of treatment is available in several forms and when properly administered, has been demonstrated to assist in somewhere between 58% and 65% of cases.\",\n \"Masking is simply the addition of an outside sound that serves as a substitute or mask for the tinnitus.\",\n \"Masking systems known to date are typically worn within the ear canal or positioned nearby so as to ensure provision of a masking sound to the sufferer and as a result these devices stigmatise the wearer and are worn reluctantly.\",\n \"Implantable tinnitus maskers are known, such as that described in U.S. Pat.\",\n \"No.\",\n \"5,795,287.Such devices utilise electromechanical transducers coupled to the ossicular chain to produce the artificial masking sounds, however, these devices require a very complicated surgery to implant as the electromechanical transducer must mechanically manipulate the ossicular chain.\",\n \"Also, it has been found that such mechanical coupling is not always guaranteed to be stable as pressure necroses in the area of the middle ear ossicle has been found to occur in a number of cases resulting in bone erosion.\",\n \"The present invention relates to a new system for treating the symptoms of tinnitus that preferably does not require complicated surgery nor the fixation of electromechanical transducers to the ossicles.\",\n \"Any discussion of documents, acts, materials, devices, articles or the like which has been included in the present specification is solely for the purpose of providing a context for the present invention.\",\n \"It is not to be taken as an admission that any or all of these matters form part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed before the priority date of each claim of this application.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>Throughout this specification the word “comprise”, or variations such as “comprises” or “comprising”, will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps.\",\n \"The present invention is firstly directed to an implant that can be inserted in the cochlea but in a position external to the scala tympani.\",\n \"Such an implant provides an alternative option for those persons described above who would benefit from the use of a cochlear implant to improve hearing of relatively high frequencies but who have some residual hearing of relatively low frequencies.\",\n \"The present invention further preferably aims to provide a cochlear implant system that preserves the normal hydrodynamic nature of the cochlea allowing for an electrode array to be positioned to stimulate the desired neurons without causing damage to the important internal ducts of the cochlea.\",\n \"The present invention is secondly directed to an implant, as described above, that can be used as a means of masking the symptoms of tinnitus.\",\n \"According to a first aspect, the present application is directed to a first invention comprising an implantable tissue-stimulating device having a carrier member having at least one electrode thereon, the carrier member being adapted for intracochlear but extraluminar insertion within the cochlea of an implantee.\",\n \"According to a second aspect, the present invention is directed to a second invention comprising an implantable tissue-stimulating device for use in the masking or treatment of the symptoms of tinnitus, the device having a carrier member having at least one electrode thereon, the carrier member being adapted for intracochlear but extraluminar insertion within the cochlea of an implantee.\",\n \"According to a third aspect, the present invention is directed to a third invention comprising an implantable tissue-stimulating device when used in the masking or treatment of the symptoms of tinnitus, the device having a carrier member having at least one electrode thereon, the carrier member being adapted for intracochlear but extraluminar insertion within the cochlea of an implantee.\",\n \"In the above aspects, the device can be a cochlear implant In one embodiment, the carrier member can be a cochlear implant carrier member.\",\n \"The carrier member preferably has a body having a plurality of electrodes mounted thereon.\",\n \"The electrodes can be disposed in an array on the carrier member.\",\n \"The electrodes can be adapted to apply a preselected tissue stimulation.\",\n \"In a preferred embodiment, the carrier member is adapted to be implanted in a crevice between the spiral ligament and the endosteum of the lateral wall of the cochlea.\",\n \"This is quite different to the normal placement of the electrode array of a traditional cochlear implant in the scala tympani of the cochlea.\",\n \"The placement of the device is preferably designed to avoid any breach of the internal ducts of the cochlea (eg.\",\n \"scala tympani and scala vestibule) so that the normal hydrodynamic behaviour of the cochlea is not affected by any intrusive device.\",\n \"This is important, as for implantees suffering tinnitus, use of the device does not lead to loss of what otherwise may be good hearing.\",\n \"For implantees with at least some sensorineural hearing loss, use of the device maximises the possibility of also preserving residual hearing offered by the implantee's cochlea.\",\n \"In this case, it is envisaged that use of the device will have particular benefit in those instances where the implantee has substantial residual hearing in the low frequencies but would benefit from supplemental stimulation in a relatively higher frequency range.\",\n \"In this case, the implantee may benefit from use of a hearing aid that amplifies the relatively low frequencies still detectable by the implantee and a cochlear implant for detection of relatively high frequencies.\",\n \"In a preferred embodiment, the carrier member has a maximum length of about 7-10 mm, a width of about 0.6 mm and a thickness no greater than about 0.2 mm and, more preferably, about 0.1 mm.\",\n \"In a further embodiment, the carrier member can have an inner surface and an outer surface, the inner surface being adapted to face inwardly into the cochlea, while the outer surface faces toward the endosteum of the cochlea.\",\n \"In one embodiment, the inner face can have a concavity.\",\n \"In a further embodiment, the outer face can have a convexity.\",\n \"In a further embodiment, the thickness of the carrier member between its inner surface and outer surface can be substantially constant for at least a majority of its length from the proximal end to the distal end.\",\n \"In another embodiment, the thickness of the carrier can change, such as decrease, from the proximal end to the distal end.\",\n \"In a preferred embodiment, the carrier can be relatively more resiliently flexible in a longitudinal plane and relatively less resiliently flexible in a lateral plane.\",\n \"The carrier member can be relatively flexible and preferably adapted to follow the curvature of the endosteum along the basal turn.\",\n \"In a preferred embodiment, a proximal end of the carrier member can be identified by a stop member.\",\n \"The stop member can extend substantially at right angles to the longitudinal axis of the carrier member.\",\n \"The stop member preferably has a length of between about 1.5 and 2.0 mm.\",\n \"The stop member can serve as both a region for grasping the carrier member and also act to prevent insertion of the carrier member within the crevice beyond a predetermined maximum depth.\",\n \"In a still further embodiment, said at least one electrode has a surface that is at least adjacent the inner surface of the carrier.\",\n \"More preferably, each of the electrodes in the array has a surface that is adjacent the inner surface of the carrier member.\",\n \"In a further embodiment, the surfaces of the electrodes are aligned with the inner surface of the carrier member.\",\n \"In another embodiment, the surfaces of the electrodes stand proud of the inner surface of the carrier member.\",\n \"It is also envisaged that the electrode surface could also be recessed into the inner surface of the carrier member.\",\n \"In one embodiment, the carrier member can be formed from a biocompatible elastomeric material.\",\n \"In one embodiment, the elastomeric material can be a silicone rubber.\",\n \"In another embodiment, the carrier member can be formed from a biocompatible polyurethane or similar material.\",\n \"The surfaces of the carrier member are preferably smooth to prevent any damage to the cochlea as the array is placed in the cochlea.\",\n \"In a preferred embodiment, the electrode array can include electrically conducting wires connected to the electrodes and extending to at least said proximal end.\",\n \"In one embodiment, one wire can be connected to each of said electrodes.\",\n \"In another embodiment, at least two wires can be connected to each of said electrodes.\",\n \"Each electrode can comprise a contact element.\",\n \"The carrier member can have a longitudinal axis with each contact element arranged orthogonally to the longitudinal axis.\",\n \"The contact elements can be formed from a biocompatible material.\",\n \"The biocompatible material of the contact element can be platinum.\",\n \"The wires may preferably be connected to the contact elements by welding, or any other suitable connecting method.\",\n \"Once implanted, the electrodes of the carrier member preferably receive stimulation signals from a stimulator means.\",\n \"The stimulator means is preferably electrically connected to the carrier member by way of an electrical lead.\",\n \"The lead can include the one or more wires extending from each electrode of the array mounted on the carrier member.\",\n \"In one embodiment, the lead can extend from the carrier member to the stimulator means or at least the housing thereof.\",\n \"In one embodiment, the lead is continuous with no electrical connectors, at least external the housing of the.\",\n \"stimulator means, required to connect the wires extending from the electrodes to the stimulator means.\",\n \"One advantage of this arrangement is that there is no requirement for the surgeon implanting the device to make the necessary electrical connection between the wires extending from the electrodes and the stimulator means.\",\n \"The stimulator means is preferably positioned within a housing that is implantable within the implantee.\",\n \"The housing for the stimulator means is preferably implantable within a recess in the bone behind the ear posterior to the mastoid.\",\n \"When implanted, the housing preferably contains, in addition to the stimulator means, a receiver means.\",\n \"The receiver means is preferably adapted to receive signals from a controller means.\",\n \"The controller means is, in use, preferably mounted external to the body of the implantee such that the signals are transmitted transcutaneously through the skin of the implantee.\",\n \"Signals can preferably travel from the controller means to the receiver means and vice versa.\",\n \"The receiver means can include a receiver coil adapted to receive radio frequency (RF) signals from a corresponding transmitter coil worn externally of the body.\",\n \"The radio frequency signals can comprise frequency modulated (FM) signals.\",\n \"While described as a receiver coil, the receiver coil can preferably transmit signals to the transmitter coil which receives the signals.\",\n \"The transmitter coil is preferably held in position adjacent the implanted location of the receiver coil by way of respective attractive magnets mounted centrally in, or at some other position relative to, the coils.\",\n \"The external controller can comprise a processor adapted to output one or more stimulation regimes to the stimulator.\",\n \"Where the device is being used to mask or treat the symptoms of tinnitus, the external controller can comprise a processor adapted to output one or more stimulation regimes to the stimulator.\",\n \"In one embodiment of this application, the stimulation regime can comprise a random continuous sub-threshold stimulation regime.\",\n \"In this regime, the stimulation signals output to the electrodes of the carrier member are at a level below the threshold of hearing of the sufferer.\",\n \"In another embodiment, the stimulation regime can comprise a random continuous supra-threshold stimulation, such as white noise.\",\n \"In a still further embodiment, the stimulation regime can comprise a random discontinuous supra-threshold stimulation regime.\",\n \"It is postulated by the present inventors that irregular stimulation may be sufficient to reduce the impact of the tinnitus condition.\",\n \"Irregular stimulation also has the advantage of being relatively power-efficient and hence would result in longer battery life for the device.\",\n \"In yet a further embodiment, the stimulation regime can comprise a treatment-on-demand regime.\",\n \"Such a regime is postulated by the present inventors as being advantageous for those persons who only suffer irregular episodes of tinnitus.\",\n \"In this embodiment, the external controller can further comprise an activation means.\",\n \"The activation means can comprise a switch means on the external controller.\",\n \"In this case, the sufferer or a third person could activate the processor when required.\",\n \"Where the device is being used to provide a hearing sensation, the external controller preferably comprises a speech processor adapted to receive signals output by a microphone.\",\n \"During use, the microphone is preferably worn on the pinna of the implanted, however, other suitable locations can be envisaged, such as the lapel of the implantee's clothing.\",\n \"The speech processor encodes the sound detected by the microphone into a sequence of electrical stimuli following given algorithms, such as algorithms already developed for cochlear implant systems.\",\n \"The encoded sequence is transferred to the implanted receiver/stimulator means using the transmitter and receiver coils.\",\n \"The implanted receiver/stimulator means demodulates the modulated FM signal and allocates the electrical pulses to the appropriate attached electrode by an algorithm which is consistent with the chosen speech coding strategy.\",\n \"In one embodiment, the processor can be adapted to receive signals from the microphone when external sounds, such as speech, are present and to output a stimulation regime, when no external sounds are present, that is adapted to mask or treat the symptoms of tinnitus.\",\n \"The external controller preferably further comprises a power supply.\",\n \"The power supply can comprise one or more rechargeable batteries.\",\n \"The transmitter and receiver coils are used to provide power via transcutaneous induction to the implanted receiver/stimulator means and the electrode array.\",\n \"While the implant system can rely on external componentry, in another embodiment, the controller means, including the microphone where present, the processor, and the power supply can also be implantable.\",\n \"In this embodiment, the controller means can be contained within a hermetically sealed housing or the housing used for the stimulator means.\",\n \"An implantable controller means also preferably has an activation means that provides the implantee or a third person with a means to activate a stimulation regime when required.\",\n \"In one embodiment, the inactivation means can comprise a magnetic switch.\",\n \"In this case, the implantable controller means would preferably incorporate a magnetic field detector which is triggered on detecting the presence of a suitable magnet held dose to the location of the implantable controller.\",\n \"In another embodiment, the activation means can comprise a radio frequency switch means.\",\n \"In this case, the implantable controller can include a radio frequency detector means adapted to receive a particular predetermined or programmed signal from a radio transmitter.\",\n \"The radio transmitter is activated when required by the implantee or a third person.\",\n \"In yet another embodiment, the activation means can comprise an infrared switch means.\",\n \"In this case, the implantable controller can include a infrared detector means adapted to receive a particular pre-determined or programmed infrared signal from a infrared transmitter.\",\n \"The infrared transmitter is activated when required by the implantee or a third person.\",\n \"According to a fourth aspect, the present invention is directed to a fourth invention comprising a method of inserting a tissue-stimulating device as defined above into a cochlea of an implantee, the method comprising the steps of: performing a fenestration to access the crevice of the cochlea between the spiral ligament and the endosteum of the lateral wall of the cochlea; placing the device in the crevice; and closing the fenestration.\",\n \"According to a fifth aspect, the present invention is directed to a fifth invention comprising a method of treating the symptoms of tinnitus comprising inserting a device as defined above into a cochlea of an implantee, the method comprising the steps of: performing a fenestration to access the crevice of the cochlea between the spiral ligament and the endosteum of the lateral wall of the cochlea; placing the device in the crevice; and closing the fenestration.\",\n \"In a preferred embodiment of the fourth and fifth aspects, the device is guided very gently through the fenestration and into the crevice.\",\n \"Following placement of the device, the method can include the step of covering the fenestration with soft body tissue and/or bony dust mixed with fibrin.\",\n \"Once in place, the device can be used to output-one or more stimulation regimes to the cochlea to provide a hearing sensation and/or mask or treat the symptoms of tinnitus as described above.\"\n ],\n [\n \"FIELD OF THE INVENTION The present invention relates to an auditory prosthesis adapted for intracochlear but extraluminar placement within the cochlea.\",\n \"An application of the device for masking or treating tinnitus is also described.\",\n \"BACKGROUND OF THE INVENTION Hearing loss, which may be due to many different causes, is generally of two types, conductive and sensorineural.\",\n \"In some cases, a person may have hearing loss of both types.\",\n \"Of them, conductive hearing loss occurs where the normal mechanical pathways for sound to reach the hair cells in the cochlea are impeded, for example, by damage to the ossicdes.\",\n \"Conductive hearing loss may often be helped by use of conventional hearing aids, which amplify sound so that acoustic information does reach the cochlea and the hair cells.\",\n \"In many people who are profoundly deaf, however, the reason for their deafness is sensorineural hearing loss.\",\n \"This type of hearing loss is due to the absence of, or destruction of, the hair cells in the cochlea which transduce acoustic signals into nerve impulses.\",\n \"These people are thus unable to derive suitable benefit from conventional hearing aid systems, no matter how loud the acoustic stimulus is made, because there is damage to or absence of the mechanism for nerve impulses to be generated from sound in the normal manner.\",\n \"It is for this purpose that cochlear implant systems have been developed.\",\n \"Such systems bypass the hair cells in the cochlea and directly deliver electrical stimulation to the auditory nerve fibres, thereby allowing the brain to perceive a hearing sensation resembling the natural hearing sensation normally delivered to the auditory nerve.\",\n \"U.S. Pat.\",\n \"No.\",\n \"4,532,930, the contents of which are incorporated herein by reference, provides a description of one type of traditional cochlear implant system.\",\n \"Typically, cochlear implant systems have consisted of essentially two components, an external component commonly referred to as a processor unit and an internal implanted component commonly referred to as a receiver/stimulator unit.\",\n \"Traditionally, both of these components have cooperated together to provide the sound sensation to a user.\",\n \"The external component has traditionally consisted of a microphone for detecting sounds, such as speech and environmental sounds, a speech processor that converts speech into a coded signal, a power source such as a battery, and an external transmitter coil.\",\n \"The coded signal output by the sound processor is transmitted transcutaneously to the implanted receiver/stimulator unit situated within a recess of the temporal bone of the user.\",\n \"This transcutaneous transmission occurs via the external transmitter coil which is positioned to communicate with an implanted receiver coil provided with the stimulator/receiver unit.\",\n \"This communication serves two essential purposes, firstly to transcutaneously transmit the coded sound signal and secondly to provide power to the implanted receiver/stimulator unit Conventionally, this link has been in the form of a radio frequency (RF) link, but other such links have been proposed and implemented with varying degrees of success.\",\n \"The implanted receiver/stimulator unit traditionally includes a receiver coil that receives the coded signal and power from the external processor component, and a stimulator that processes the coded signal and outputs a stimulation signal to an intracochlea electrode assembly which applies the electrical stimulation directly to the auditory nerve producing a hearing sensation corresponding to the original detected sound.\",\n \"Traditionally, at least the speech processor of the external componentry has been carried on the body of the user, such as in a pocket of the user's clothing, a belt pouch or in a harness, while the microphone has been mounted on a clip mounted behind the ear or on the lapel of the user.\",\n \"More recently, due in the main to improvements in technology, the physical dimensions of the sound processor have been able to be reduced allowing for the external componentry to be housed in a small unit capable of being worn behind the ear of the user.\",\n \"This unit allows the microphone, power unit and the sound processor to be housed in a single unit capable of being discretely worn behind the ear, with the external transmitter coil still positioned on the side of the user's head to allow for the transmission of the coded sound signal from the sound processor and power to the implanted stimulator unit.\",\n \"It is further envisaged that with continual improvements in technology all the traditional external componentry may be implanted in the user.\",\n \"In such a system all of the speech processing may be performed inside the implanted stimulator unit, via an implanted microphone.\",\n \"It is known in the art that the cochlea is tonotopically mapped.\",\n \"In other words, the cochlea can be partitioned into regions, with each region being responsive to signals in a particular frequency range.\",\n \"This property of the cochlea has been exploited by providing the electrode assembly with an array of electrodes, each electrode being arranged and constructed to deliver a stimulating signal within a preselected frequency range, to the appropriate region within the scala tympani of the cochlea.\",\n \"The electrical currents and electric fields from each electrode stimulate the nerves disposed on the modiolus of the cochlea.\",\n \"Despite the enormous benefits offered by cochlear implants, one potential disadvantage of placement of the electrode assembly within the scala tympani is that it is necessary to breach the internal ducts of the cochlea, generally the scala tympani.\",\n \"The breaching of the scala tympani of the cochlea adversely affects the hydrodynamic behaviour of the cochlea and is thought to prevent or at least reduce any chance of preservation of any residual hearing of the implantee.\",\n \"This can be problematic for those persons who would benefit from use of a cochlear implant to improve hearing of relatively high frequencies but who have some residual hearing of relatively low frequencies.\",\n \"In such a case, the implantee is forced to trade off an existing residual capacity to hear relatively low frequency sounds against the desirability of being able to have a hearing sensation of relatively high frequency sounds offered by a cochlear implant.\",\n \"There have been a number of proposals put forward to provide a hybrid system whereby a cochlear implant system can be used in conjunction with residual hearing, usually assisted by the use of a hearing aid.\",\n \"One such example of a proposed system is described in International Patent Application No WO 00/69512.In this application, the hybrid system utilises a hearing aid to amplify the low frequency sound enabling the user to rely on normal hearing processes to experience such sounds.\",\n \"For high frequency sounds, the hybrid system utilises a relatively conventional cochlear stimulation device consisting of a short cochlear electrode array.\",\n \"The short cochlear electrode array of this application is described as consisting of 4-8 electrodes and is inserted directly through the round window membrane making contact with the basal region of the cochlea.\",\n \"Therefore the system as described in this application still uses a relatively obtrusive electrode array making it very difficult to preserve any residual hearing the patient may have in such areas.\",\n \"As already described, the present application is also directed to a device for masking or treating tinnitus.\",\n \"Tinnitus is the medical term for a condition in which sufferers report a ringing in their ears or head when there is in fact no external sound present in the sufferer's audible range.\",\n \"Although some people hear a ringing noise, others report the noise as being a hissing, a chirping, or a clicking.\",\n \"There are various estimates as to how many sufferers of tinnitus there are worldwide.\",\n \"For example, it is suggested that some 50 million Americans suffer from tinnitus, with about 83% of them hearing a constant ringing.\",\n \"Other figures suggest at least 12 million people have tinnitus to what is regarded as a distressing degree.\",\n \"For some people, tinnitus is just a nuisance.\",\n \"For others, it can be a quite debilitating condition.\",\n \"Usually, the only relief tinnitus sufferers will experience is an occasional reduction in the loudness of the tinnitus from time to time.\",\n \"The cause of tinnitus or at least its onset is unclear.\",\n \"There is, however, data available that demonstrates that exposure to loud noise is a trigger for the condition.\",\n \"Other suggested triggers include severe head trauma, certain medications, sinus and respiratory infections, ear infections, wax build-up and certain types of tumours.\",\n \"There are, as yet, no cures for tinnitus but there are several treatments currently used to provide at least some relief.\",\n \"One treatment is the use of what are commonly referred to as tinnitus maskers.\",\n \"One example of a tinnitus masker is disclosed in PCT Patent Application WO 90/07251.Tinnitus maskers are essentially small battery operated devices which are worn like a hearing aid behind or in the ear, and cover (mask) the tinnitus psychoacoustically by artificial sounds which are emitted, for example, via a hearing aid speaker into the auditory canal and which reduce the disturbing tinnitus as far as possible below the threshold of perception.\",\n \"The artificial sounds are often narrowband noise (for example, third octave noise) which in its spectral position and its loudness level can be adjusted via a programming device to enable the maximum position adaptation to the individual tinnitus situation.\",\n \"This form of treatment is available in several forms and when properly administered, has been demonstrated to assist in somewhere between 58% and 65% of cases.\",\n \"Masking is simply the addition of an outside sound that serves as a substitute or mask for the tinnitus.\",\n \"Masking systems known to date are typically worn within the ear canal or positioned nearby so as to ensure provision of a masking sound to the sufferer and as a result these devices stigmatise the wearer and are worn reluctantly.\",\n \"Implantable tinnitus maskers are known, such as that described in U.S. Pat.\",\n \"No.\",\n \"5,795,287.Such devices utilise electromechanical transducers coupled to the ossicular chain to produce the artificial masking sounds, however, these devices require a very complicated surgery to implant as the electromechanical transducer must mechanically manipulate the ossicular chain.\",\n \"Also, it has been found that such mechanical coupling is not always guaranteed to be stable as pressure necroses in the area of the middle ear ossicle has been found to occur in a number of cases resulting in bone erosion.\",\n \"The present invention relates to a new system for treating the symptoms of tinnitus that preferably does not require complicated surgery nor the fixation of electromechanical transducers to the ossicles.\",\n \"Any discussion of documents, acts, materials, devices, articles or the like which has been included in the present specification is solely for the purpose of providing a context for the present invention.\",\n \"It is not to be taken as an admission that any or all of these matters form part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed before the priority date of each claim of this application.\",\n \"SUMMARY OF THE INVENTION Throughout this specification the word “comprise”, or variations such as “comprises” or “comprising”, will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps.\",\n \"The present invention is firstly directed to an implant that can be inserted in the cochlea but in a position external to the scala tympani.\",\n \"Such an implant provides an alternative option for those persons described above who would benefit from the use of a cochlear implant to improve hearing of relatively high frequencies but who have some residual hearing of relatively low frequencies.\",\n \"The present invention further preferably aims to provide a cochlear implant system that preserves the normal hydrodynamic nature of the cochlea allowing for an electrode array to be positioned to stimulate the desired neurons without causing damage to the important internal ducts of the cochlea.\",\n \"The present invention is secondly directed to an implant, as described above, that can be used as a means of masking the symptoms of tinnitus.\",\n \"According to a first aspect, the present application is directed to a first invention comprising an implantable tissue-stimulating device having a carrier member having at least one electrode thereon, the carrier member being adapted for intracochlear but extraluminar insertion within the cochlea of an implantee.\",\n \"According to a second aspect, the present invention is directed to a second invention comprising an implantable tissue-stimulating device for use in the masking or treatment of the symptoms of tinnitus, the device having a carrier member having at least one electrode thereon, the carrier member being adapted for intracochlear but extraluminar insertion within the cochlea of an implantee.\",\n \"According to a third aspect, the present invention is directed to a third invention comprising an implantable tissue-stimulating device when used in the masking or treatment of the symptoms of tinnitus, the device having a carrier member having at least one electrode thereon, the carrier member being adapted for intracochlear but extraluminar insertion within the cochlea of an implantee.\",\n \"In the above aspects, the device can be a cochlear implant In one embodiment, the carrier member can be a cochlear implant carrier member.\",\n \"The carrier member preferably has a body having a plurality of electrodes mounted thereon.\",\n \"The electrodes can be disposed in an array on the carrier member.\",\n \"The electrodes can be adapted to apply a preselected tissue stimulation.\",\n \"In a preferred embodiment, the carrier member is adapted to be implanted in a crevice between the spiral ligament and the endosteum of the lateral wall of the cochlea.\",\n \"This is quite different to the normal placement of the electrode array of a traditional cochlear implant in the scala tympani of the cochlea.\",\n \"The placement of the device is preferably designed to avoid any breach of the internal ducts of the cochlea (eg.\",\n \"scala tympani and scala vestibule) so that the normal hydrodynamic behaviour of the cochlea is not affected by any intrusive device.\",\n \"This is important, as for implantees suffering tinnitus, use of the device does not lead to loss of what otherwise may be good hearing.\",\n \"For implantees with at least some sensorineural hearing loss, use of the device maximises the possibility of also preserving residual hearing offered by the implantee's cochlea.\",\n \"In this case, it is envisaged that use of the device will have particular benefit in those instances where the implantee has substantial residual hearing in the low frequencies but would benefit from supplemental stimulation in a relatively higher frequency range.\",\n \"In this case, the implantee may benefit from use of a hearing aid that amplifies the relatively low frequencies still detectable by the implantee and a cochlear implant for detection of relatively high frequencies.\",\n \"In a preferred embodiment, the carrier member has a maximum length of about 7-10 mm, a width of about 0.6 mm and a thickness no greater than about 0.2 mm and, more preferably, about 0.1 mm.\",\n \"In a further embodiment, the carrier member can have an inner surface and an outer surface, the inner surface being adapted to face inwardly into the cochlea, while the outer surface faces toward the endosteum of the cochlea.\",\n \"In one embodiment, the inner face can have a concavity.\",\n \"In a further embodiment, the outer face can have a convexity.\",\n \"In a further embodiment, the thickness of the carrier member between its inner surface and outer surface can be substantially constant for at least a majority of its length from the proximal end to the distal end.\",\n \"In another embodiment, the thickness of the carrier can change, such as decrease, from the proximal end to the distal end.\",\n \"In a preferred embodiment, the carrier can be relatively more resiliently flexible in a longitudinal plane and relatively less resiliently flexible in a lateral plane.\",\n \"The carrier member can be relatively flexible and preferably adapted to follow the curvature of the endosteum along the basal turn.\",\n \"In a preferred embodiment, a proximal end of the carrier member can be identified by a stop member.\",\n \"The stop member can extend substantially at right angles to the longitudinal axis of the carrier member.\",\n \"The stop member preferably has a length of between about 1.5 and 2.0 mm.\",\n \"The stop member can serve as both a region for grasping the carrier member and also act to prevent insertion of the carrier member within the crevice beyond a predetermined maximum depth.\",\n \"In a still further embodiment, said at least one electrode has a surface that is at least adjacent the inner surface of the carrier.\",\n \"More preferably, each of the electrodes in the array has a surface that is adjacent the inner surface of the carrier member.\",\n \"In a further embodiment, the surfaces of the electrodes are aligned with the inner surface of the carrier member.\",\n \"In another embodiment, the surfaces of the electrodes stand proud of the inner surface of the carrier member.\",\n \"It is also envisaged that the electrode surface could also be recessed into the inner surface of the carrier member.\",\n \"In one embodiment, the carrier member can be formed from a biocompatible elastomeric material.\",\n \"In one embodiment, the elastomeric material can be a silicone rubber.\",\n \"In another embodiment, the carrier member can be formed from a biocompatible polyurethane or similar material.\",\n \"The surfaces of the carrier member are preferably smooth to prevent any damage to the cochlea as the array is placed in the cochlea.\",\n \"In a preferred embodiment, the electrode array can include electrically conducting wires connected to the electrodes and extending to at least said proximal end.\",\n \"In one embodiment, one wire can be connected to each of said electrodes.\",\n \"In another embodiment, at least two wires can be connected to each of said electrodes.\",\n \"Each electrode can comprise a contact element.\",\n \"The carrier member can have a longitudinal axis with each contact element arranged orthogonally to the longitudinal axis.\",\n \"The contact elements can be formed from a biocompatible material.\",\n \"The biocompatible material of the contact element can be platinum.\",\n \"The wires may preferably be connected to the contact elements by welding, or any other suitable connecting method.\",\n \"Once implanted, the electrodes of the carrier member preferably receive stimulation signals from a stimulator means.\",\n \"The stimulator means is preferably electrically connected to the carrier member by way of an electrical lead.\",\n \"The lead can include the one or more wires extending from each electrode of the array mounted on the carrier member.\",\n \"In one embodiment, the lead can extend from the carrier member to the stimulator means or at least the housing thereof.\",\n \"In one embodiment, the lead is continuous with no electrical connectors, at least external the housing of the.\",\n \"stimulator means, required to connect the wires extending from the electrodes to the stimulator means.\",\n \"One advantage of this arrangement is that there is no requirement for the surgeon implanting the device to make the necessary electrical connection between the wires extending from the electrodes and the stimulator means.\",\n \"The stimulator means is preferably positioned within a housing that is implantable within the implantee.\",\n \"The housing for the stimulator means is preferably implantable within a recess in the bone behind the ear posterior to the mastoid.\",\n \"When implanted, the housing preferably contains, in addition to the stimulator means, a receiver means.\",\n \"The receiver means is preferably adapted to receive signals from a controller means.\",\n \"The controller means is, in use, preferably mounted external to the body of the implantee such that the signals are transmitted transcutaneously through the skin of the implantee.\",\n \"Signals can preferably travel from the controller means to the receiver means and vice versa.\",\n \"The receiver means can include a receiver coil adapted to receive radio frequency (RF) signals from a corresponding transmitter coil worn externally of the body.\",\n \"The radio frequency signals can comprise frequency modulated (FM) signals.\",\n \"While described as a receiver coil, the receiver coil can preferably transmit signals to the transmitter coil which receives the signals.\",\n \"The transmitter coil is preferably held in position adjacent the implanted location of the receiver coil by way of respective attractive magnets mounted centrally in, or at some other position relative to, the coils.\",\n \"The external controller can comprise a processor adapted to output one or more stimulation regimes to the stimulator.\",\n \"Where the device is being used to mask or treat the symptoms of tinnitus, the external controller can comprise a processor adapted to output one or more stimulation regimes to the stimulator.\",\n \"In one embodiment of this application, the stimulation regime can comprise a random continuous sub-threshold stimulation regime.\",\n \"In this regime, the stimulation signals output to the electrodes of the carrier member are at a level below the threshold of hearing of the sufferer.\",\n \"In another embodiment, the stimulation regime can comprise a random continuous supra-threshold stimulation, such as white noise.\",\n \"In a still further embodiment, the stimulation regime can comprise a random discontinuous supra-threshold stimulation regime.\",\n \"It is postulated by the present inventors that irregular stimulation may be sufficient to reduce the impact of the tinnitus condition.\",\n \"Irregular stimulation also has the advantage of being relatively power-efficient and hence would result in longer battery life for the device.\",\n \"In yet a further embodiment, the stimulation regime can comprise a treatment-on-demand regime.\",\n \"Such a regime is postulated by the present inventors as being advantageous for those persons who only suffer irregular episodes of tinnitus.\",\n \"In this embodiment, the external controller can further comprise an activation means.\",\n \"The activation means can comprise a switch means on the external controller.\",\n \"In this case, the sufferer or a third person could activate the processor when required.\",\n \"Where the device is being used to provide a hearing sensation, the external controller preferably comprises a speech processor adapted to receive signals output by a microphone.\",\n \"During use, the microphone is preferably worn on the pinna of the implanted, however, other suitable locations can be envisaged, such as the lapel of the implantee's clothing.\",\n \"The speech processor encodes the sound detected by the microphone into a sequence of electrical stimuli following given algorithms, such as algorithms already developed for cochlear implant systems.\",\n \"The encoded sequence is transferred to the implanted receiver/stimulator means using the transmitter and receiver coils.\",\n \"The implanted receiver/stimulator means demodulates the modulated FM signal and allocates the electrical pulses to the appropriate attached electrode by an algorithm which is consistent with the chosen speech coding strategy.\",\n \"In one embodiment, the processor can be adapted to receive signals from the microphone when external sounds, such as speech, are present and to output a stimulation regime, when no external sounds are present, that is adapted to mask or treat the symptoms of tinnitus.\",\n \"The external controller preferably further comprises a power supply.\",\n \"The power supply can comprise one or more rechargeable batteries.\",\n \"The transmitter and receiver coils are used to provide power via transcutaneous induction to the implanted receiver/stimulator means and the electrode array.\",\n \"While the implant system can rely on external componentry, in another embodiment, the controller means, including the microphone where present, the processor, and the power supply can also be implantable.\",\n \"In this embodiment, the controller means can be contained within a hermetically sealed housing or the housing used for the stimulator means.\",\n \"An implantable controller means also preferably has an activation means that provides the implantee or a third person with a means to activate a stimulation regime when required.\",\n \"In one embodiment, the inactivation means can comprise a magnetic switch.\",\n \"In this case, the implantable controller means would preferably incorporate a magnetic field detector which is triggered on detecting the presence of a suitable magnet held dose to the location of the implantable controller.\",\n \"In another embodiment, the activation means can comprise a radio frequency switch means.\",\n \"In this case, the implantable controller can include a radio frequency detector means adapted to receive a particular predetermined or programmed signal from a radio transmitter.\",\n \"The radio transmitter is activated when required by the implantee or a third person.\",\n \"In yet another embodiment, the activation means can comprise an infrared switch means.\",\n \"In this case, the implantable controller can include a infrared detector means adapted to receive a particular pre-determined or programmed infrared signal from a infrared transmitter.\",\n \"The infrared transmitter is activated when required by the implantee or a third person.\",\n \"According to a fourth aspect, the present invention is directed to a fourth invention comprising a method of inserting a tissue-stimulating device as defined above into a cochlea of an implantee, the method comprising the steps of: performing a fenestration to access the crevice of the cochlea between the spiral ligament and the endosteum of the lateral wall of the cochlea; placing the device in the crevice; and closing the fenestration.\",\n \"According to a fifth aspect, the present invention is directed to a fifth invention comprising a method of treating the symptoms of tinnitus comprising inserting a device as defined above into a cochlea of an implantee, the method comprising the steps of: performing a fenestration to access the crevice of the cochlea between the spiral ligament and the endosteum of the lateral wall of the cochlea; placing the device in the crevice; and closing the fenestration.\",\n \"In a preferred embodiment of the fourth and fifth aspects, the device is guided very gently through the fenestration and into the crevice.\",\n \"Following placement of the device, the method can include the step of covering the fenestration with soft body tissue and/or bony dust mixed with fibrin.\",\n \"Once in place, the device can be used to output-one or more stimulation regimes to the cochlea to provide a hearing sensation and/or mask or treat the symptoms of tinnitus as described above.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS By way of example only, a preferred mode of carrying out the invention is described with reference to the accompanying drawings, in which: FIG.\",\n \"1 is a perspective view of one embodiment of a carrier member of an implantable electrode array according to the present invention;: FIG.\",\n \"2 is a cross-sectional view through a human cochlea depicting the desired position for placement of the electrode array according to the present invention; and FIG.\",\n \"3 is a pictorial representation of one embodiment of a system that can utilise the carrier member of FIG.\",\n \"1 whether it be for providing a hearing sensation to an implantee and/or to mask or treat the symptoms of tinnitus.\",\n \"PREFERRED MODE OF CARRYING OUT THE INVENTION One embodiment of an electrode assembly for use in the present invention according to the present invention is depicted generally as 10 in the drawings.\",\n \"The assembly 10 includes an elongate electrode carrier member 11.For the purposes of clarity, the plurality of electrodes that are mounted on the carrier member 11 are not depicted in the drawings.\",\n \"While not depicted, the electrodes can be disposed in a linear array on the carrier member 11 and be adapted to apply a preselected tissue stimulation to the cochlea.\",\n \"The depicted carrier member 11 is preformed from a resiliently flexible biocompatible silicone and extends from a distal end 12 to a stop member 13.The carrier member 11 is adapted for intracochlear but extraluminar insertion within the cochlea of an implantee.\",\n \"In use, the carrier member 11 is adapted to be implanted in the crevice 21 (see FIG.\",\n \"2) between the spiral ligament 22 and the endosteum 23 of the lateral wall of the cochlea 20.This is a quite different location to the normal placement of the cochlear implant electrode array in the scala tympani 24 of the cochlea 20.The placement of the carrier member 11 is designed to avoid any breach of the internal ducts of the cochlea 20 (eg.\",\n \"scala tympani 24 and scala vestibuli 25) so that the normal hydrodynamic behaviour of the cochlea 20 is not affected by any intrusive device.\",\n \"By preserving the normal hydrodynamic behaviour of the cochlea 20, use of the carrier member 11 maximises the possibility of also preserving any hearing of the implantee that is offered by the cochlea 20.In the depicted embodiment, the carrier member 11 has a maximum length of about 8-10 mm, a width of about 0.6 mm and a thickness no greater than about 0.1 mm.\",\n \"The thickness of the carrier member between its inner surface and outer surface is substantially constant for at least a majority of its length from the distal end 12 to the stop member 13, except in the region adjacent the end 12 where the thickness gradually tapers towards end 12.The depicted carrier 11 is more resiliently flexible in a longitudinal plane and relatively less resiliently flexible in a lateral plane.\",\n \"The carrier member 11 is adapted to follow the curvature of the endosteum along the basal turn of the cochlea 20.As depicted, the stop member 13 extends substantially at right angles to the longitudinal axis of the carrier member 11.The depicted stop member 13 has a length of between about 1.5 and 2.0 mm.\",\n \"The stop member 13 serves as both a region for grasping the carrier member 11 and also acts to prevent insertion of the carrier member 11 within the crevice 21 beyond a predetermined maximum depth (eg.\",\n \"8-10 mm).\",\n \"While not depicted, the electrode assembly 10 includes electrically conducting wires connected to the electrodes and extending at least beyond the stop member 13.In the depicted embodiment, one wire can be connected to each of said electrodes.\",\n \"In use, a surgeon would perform a fenestration to access the crevice 21 between the spiral ligament 22 and the endosteum 23 of the lateral wall of the cochlea 20.The carrier member 11 would then be inserted into the crevice 21 using an insertion tool that grips the stop member 13 and guides the carrier member 11 very gently into the crevice 21.Once positioned, the surgeon could close the fenestration by covering it with soft body tissue and/or bony dust mixed with fibrin.\",\n \"Once implanted, the electrodes of the carrier member 11 receive stimulation signals from an implantable stimulator 30.The stimulator 30 is positioned within a housing 31 that is implantable within the implantee.\",\n \"The housing 31 for the stimulator 30 is implantable within a recess in the bone behind the ear posterior to the mastoid.\",\n \"When implanted, the housing 31 contains, in addition to the stimulator 30, a receiver.\",\n \"The receiver is adapted to receive signals from a controller 32.The controller 32 is, in use, typically mounted external to the body 33 of the implantee such that the signals are transmitted transcutaneously through the skin of the implantee.\",\n \"The controller 32 can be worn on the body of the user or can be adapted to be worn behind the ear of the user in much the same way as conventional hearing aid devices.\",\n \"The signals travel from the controller 32 to the receiver and vice versa by use of a receiver coil 34 adapted to receive radio frequency (RF) signals from a corresponding transmitter coil 35 worn externally of the body 33.The radio frequency signals can comprise frequency modulated (FM) signals.\",\n \"While described as a receiver coil, the receiver coil 34 can also transmit signals to the transmitter coil 35 which receives the signals.\",\n \"The transmitter coil 35 is held in position adjacent the implanted location of the receiver coil 34 by way of respective attractive magnets mounted centrally in, or at some other position relative to, the coils.\",\n \"The external controller 32 comprises a processor adapted to output one or more stimulation regimes to the stimulator 31.It is also envisaged that the stimulator 31 may have the capability to store one or more stimulation regimes and upon request deliver the regime to the user via the electrodes.\",\n \"In one use of the system, the stimulation regimes of the stimulator 31 are designed to mask or treat the symptoms of tinnitus.\",\n \"In one embodiment, the stimulation regime can comprise a random continuous sub-threshold stimulation regime.\",\n \"In this regime, the stimulation signals output to the electrodes of the carrier member 11 are at level below the threshold of hearing of the sufferer.\",\n \"In another embodiment, the stimulation regime can comprise a random continuous supra-threshold stimulation, such as white noise.\",\n \"In a still further embodiment, the stimulation regime can comprise a random discontinuous supra-threshold stimulation regime.\",\n \"In yet a further embodiment, the stimulation regime can comprise a treatment-on-demand regime.\",\n \"In this embodiment, the external controller 32 can further comprise an activation means 36.The depicted activation means 36 comprises a switch on the external controller 32.In this case, the sufferer or a third person could activate the processor when required.\",\n \"In addition to the above, the processor can be adapted to receive signals output by a microphone (not depicted).\",\n \"In this case and during use, the microphone can be worn on the pinna of the implantee, however, other-suitable locations can be envisaged, such as a lapel of the implantee's clothing.\",\n \"In this case, the processor can encode the sound detected by the microphone into a sequence of electrical stimuli following given algorithms, such as algorithms already developed for cochlear implant systems.\",\n \"The encoded sequence is then transferred to the implanted stimulator 30 using the transmitter and receiver coils.\",\n \"The implanted stimulator 30 demodulates the FM signals and allocates the electrical pulses to the appropriate attached electrode by an algorithm which is consistent with the chosen speech coding strategy.\",\n \"The housing of the external controller 32 further houses a power supply.\",\n \"In the depicted embodiment, the power supply comprise one or more rechargeable batteries.\",\n \"The transmitter and receiver coils are used to provide power via transcutaneous induction to the implanted stimulator 30 and the electrode array 10.While the implant system can rely on external componentry, in another embodiment, the controller, including the microphone where present, the processor, and the power supply can also be implantable.\",\n \"In this embodiment, which is not depicted, the controller can be contained within a hermetically sealed housing or the housing used for the stimulator.\",\n \"An implantable controller means would typically have an activation means that provides the implantee or a third person with a means to activate a stimulation regime when required.\",\n \"In one embodiment, the inactivation means can comprise a magnetic switch.\",\n \"In this case, the implantable controller would preferably incorporate a magnetic field detector which is triggered on detecting the presence of a suitable magnet held close to the location of the implantable controller.\",\n \"In another embodiment, the activation means can comprise a radio frequency switch means.\",\n \"In this case, the implantable controller can include a radio frequency detector means adapted to receive a particular predetermined or programmed signal from a radio transmitter.\",\n \"The radio transmitter is activated when required by the implantee or a third person.\",\n \"In yet another embodiment, the activation means can comprise an infrared switch means.\",\n \"In this case, the implantable controller can include a infrared detector means adapted to receive a particular predetermined or programmed infrared signal from a infrared transmitter.\",\n \"The infrared transmitter is activated when required by the implantee or a third person.\",\n \"The present invention provides an implantable device that can be used to provide a hearing sensation to an implantee with sensorineural hearing loss and or mask or treat the symptoms of tinnitus while preserving the hearing of the implantee's cochlea into which the stimulating electrode array has been inserted.\",\n \"It will be appreciated by persons skilled in the art that numerous variations and/or modifications may be made to the invention as shown in the specific embodiments without departing from the spirit or scope of the invention as broadly described.\",\n \"The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473925"}}},{"rowIdx":766,"cells":{"text":{"kind":"list like","value":[["Alumina agglomerates, the preparation method thereof and use of same as an absorbent or catalyst carrier","The invention relates to alumina agglomerates of the type obtained by dehydrating an aluminium oxyhydroxide or hydroxide, agglomerating the alumina thus obtained, hydrothermally treating the agglomerates and calcinating same.","Said agglomerates are characterised in that: the V37 Å thereof is greater than or equal to 75 ml/100 g, preferably greater than or equal to 80 ml/100 g and, better still, greater than or equal to 85 ml/100 g; the V0.1 μm thereof is less than or equal to 31 ml/100 g; and the V0.2 μm thereof is less than or equal to 20 ml/100 g, preferably less than or equal to 15 ml/100 g and, better still, less than or equal to 10 ml/100 g. The invention also relates to a catalyst carrier, an intrinsic catalyst or an absorbent, in particular for use in the petroleum and petrochemical industry, comprising such alumina agglomerates.","Moreover, the invention relates to methods for preparing said agglomerates."],["1.Alumina agglomerates of the type obtained by the treatment of an aluminum hydroxide or oxyhydroxide, agglomeration of the alumina thus obtained, hydrothermal treatment of the agglomerates and calcination, wherein they have a V37 Å of greater than or equal to 75 ml/100 g, preferably greater than or equal to 80 ml/100 g and even more preferably greater than or equal to 85 ml/100 g; they have a V0.1 μm of less than or equal to 31 ml/l00 g, preferably less than or equal to 25 ml/100 g, even more preferably less than or equal to 20 ml/100 g and optimally less than or equal to 15 ml/100 g; and wherein: they have a V0.2 μm of less than or equal to 20 ml/100 g, preferably less than or equal to 15 ml/100 g and even more preferably less than or equal to 10 ml/100 g. 2.The alumina agglomerates as claimed in claim 1, wherein they have a V1 μm of less than or equal to 7 ml/100 g, preferably less than or equal to 5.5 ml/100 g and even more preferably less than or equal to 4 ml/100 g. 3.The alumina agglomerates as claimed in claim 1 wherein they have a V0.1 μm/V0.2 μm ratio of greater than or equal to 1.5, preferably greater than or equal to 2 and even more preferably greater than or equal to 2.5.4.The alumina agglomerates as claimed in claim 1 wherein they have simultaneously a V37 Å of greater than or equal to 80 ml/100 g, a V0.1 μm of less than or equal to 15 ml/100 g, a V0.2 μm of less than or equal to 10 ml/100 g, a V1 μm of less than or equal to 4 ml/100 g and a V0.1 μm/V0.2 μm ratio of greater than or equal to 2.5.5.The alumina agglomerates as claimed in claim 1 wherein they have been obtained from dehydrated hydrargillite.","6.The alumina agglomerates as claimed in claim 1 wherein they are in the form of beads.","7.The alumina agglomerates as claimed in claim 1 wherein they are in the form of extruded materials.","8.The alumina agglomerates as claimed in claim 1 wherein they are in the form of crushed materials.","9.The alumina agglomerates as claimed in claim 1 wherein they are in the form of monoliths.","10.A catalyst support, especially for the petroleum of petrochemical industry, wherein it consists of alumina agglomerates as claimed in claim 1.11.An intrinsic catalyst, especially for the petroleum or petrochemical industry, wherein it consists of alumina agglomerates as claimed in claim 1.12.An adsorbent, especially for the petroleum or petrochemical industry, wherein it consists of alumina agglomerates as claimed in claim 1.13.A process for producing alumina agglomerates as claimed in claim 6, in which: an aluminum hydroxide or oxyhydroxide, preferably hydrargillite, undergoes flash dehydration in order to obtain an active alumina powder; said active alumina powder undergoes a forming operation so as to obtain beads with a green fill density of between 500 and 1100 kg/m3, preferably between 700 and 950 kg/m3 inclusive, and a diameter predominantly between 0.8 and 10 mm, preferably between 1 and 5 mm; said beads undergo a heat treatment so as to provide them with a specific surface area of between 50 and 420 m2/g; said beads undergo a hydrothermal treatment by impregnation with water or an aqueous solution, preferably aqueous acid solution, followed by residence in an autoclave at a temperature of above 80° C.; and the agglomerates thus obtained are calcined, preferably between 500 and 1300° C. 14.The process as claimed in claim 13, wherein after the heat treatment that follows granulation, the alumina beads preferably have a V37 Å of greater than 65 ml/100 g, preferably greater than 70 ml/100 g. 15.The process as claimed in claim 13 wherein the active alumina powder obtained after dehydrating the aluminum hydroxide or oxyhydroxide is ground in order to obtain a powder with a median particle size d50 of preferably between 5 and 25 μm.","16.The process as claimed in claim 13 wherein the active alumina powder obtained after dehydrating the aluminum hydroxide or oxyhydroxide is washed with water or an aqueous acid solution.","17.The process as claimed in claim 13 wherein prior to granulation of said active alumina powder, the latter undergoes a flash operation.","18.The process for producing alumina agglomerates as claimed in claim 7, in which: an alumina-based material is mixed and extruded in order to form it; the extruded materials thus obtained are subjected to a heat treatment so as to give them a specific surface area of between 50 and 420 m2/g; said extruded materials are subjected to a hydrothermal treatment by impregnation with water or with an aqueous solution, preferably an aqueous acid solution, followed by residence in an autoclave at a temperature of above 80° C.; and the agglomerates thus obtained are calcined, preferably between 500 and 1300° C. 19.The process as claimed in claim 18, wherein said alumina-based material is dehydrated hydrargillite.","20.The process as claimed in claim 18, wherein alumina-based material comes from the precipitation of boehmite, pseudo-boehmite or bayerite, or a mixture of such materials.","21.The process as claimed in claim 13 wherein during the forming of said alumina powder or of said alumina-based material, one or more pore-forming materials that disappear on heating are added to it.","22.The process as claimed in claim 21, wherein said pore-forming materials are chosen from wood flour, charcoal, sulfur, tars, plastics or emulsions of plastics, polyvinyl alcohols, naphthalene.","23.The process as claimed in claim 13 wherein said hydrothermal treatment is carried out at a temperature of 150 to 270° C., preferably from 170 to 250° C., for a time of greater than 45 minutes, preferably from 1 to 24 hours and even more preferably from 1.5 to 12 hours.","24.The process as claimed in claim 13 wherein said hydrothermal treatment is carried out using an aqueous acid solution containing one or more mineral and/or organic acids.","25.The process as claimed in claim 24, wherein said aqueous acid solution also includes one or more compounds that can release anions capable of combining with the aluminum ions."],["The invention relates to the field of alumina agglomerates that can be used especially as adsorbents, as catalyst supports or as catalysts.","More precisely, it relates to novel alumina agglomerates having specific physical characteristics, in particular from the standpoint of their porosity, giving them particularly advantageous mechanical properties.","The invention also relates to a process for obtaining such agglomerates.","Finally, it relates to their use as catalyst supports, as catalysts or as adsorbents.","Certain catalytic processes, such as heterogeneous catalysis, for example in the field of treatment of petroleum oil cuts or the treatment of gaseous effluents, such as the exhaust gases from internal combustion engines, require the use of supports having a high porosity and good mechanical properties, such as good crush resistance and/or attrition resistance.","Alumina-based supports meet these criteria, especially when they have undergone a hydrothermal treatment, for example after the alumina has been made in the form of agglomerates.","This is because such a treatment substantially improves the mechanical properties of the agglomerates.","This hydrothermal treatment consists in impregnating the alumina agglomerates with water or an aqueous acid solution, and then heating these agglomerates, placed inside an autoclave, to a temperature of above 80° C. European Patent No.","EP-A-0 387 109, in the name of the Applicant, describes an example of an advantageous process for carrying out this hydrothermal treatment.","The agglomerates thus treated are then dried and then subjected to a heat treatment called “calcination” at a given temperature.","This calcination temperature is chosen according to the desired specific surface area and to the temperature at which these supports will be used.","The efficiency of such alumina agglomerates is to a large part dependent on their porosity: a high porosity is often favorable for the various applications envisioned, because of the lower diffusional and/or tortuosity constraints.","In applications in which the support will be used not only for satisfactorily dispersing a metallic active phase or phases supplied during production of the catalyst, but also to retain contaminants, such as during a hydrometallation operation, a high porosity is advantageous.","However, a high porosity is generally regarded as going hand in hand with a degradation of the mechanical strength of the material.","The use of alumina agglomerates with a high porosity is therefore difficult in important applications, specially in catalyst supports for which the mechanical strength of the alumina to a large extent governs the duration of use of the catalyst.","It turns out that degradation of the mechanical strength of the catalyst during use has dramatic effects, both technically and economically, on the operating process.","The object of the invention is to provide alumina agglomerates users with materials, exhibiting an excellent compromise between high porosity and mechanical strength that makes them compatible with demanding applications from the latter standpoint.","For this purpose, the subject of the invention is alumina agglomerates of the type obtained by the treatment of an aluminum hydroxide or oxyhydroxide, agglomeration of the alumina thus obtained, hydrothermal treatment of the agglomerates and calcination, characterized in that: they have a V37 Å of greater than or equal to 75 ml/100 g, preferably greater than or equal to 80 ml/100 g and even more preferably greater than or equal to 85 ml/100 g; they have a V0.1 μm of less than or equal to 31 ml/100 g, preferably less than or equal to 25 ml/100 g, even more preferably less than or equal to 20 ml/100 g and optimally less than or equal to 15 ml/100 g; and in that: they have a V0.2 μm of less than or equal to 20 ml/100 g, preferably less than or equal to 15 ml/100 g and even more preferably less than or equal to 10 ml/100 g. Preferably, they have a V1 μm of less than or equal to 7 ml/100 g, preferably less than or equal to 5.5 ml/100 g and even more preferably less than or equal to 4 ml/100 g. Preferably, they have a V0.1 μm/V0.2 μm ratio of greater than or equal to 1.5, preferably greater than or equal to 2 and even more preferably greater than or equal to 2.5.Preferably, they have simultaneously a V37 Å of greater than or equal to 80 ml/100 g, a V0.1 μm of less than or equal to 15 ml/100 g, a V0.2 μm of less than or equal to 10 ml/100 g, a V1 μm of less than or equal to 4 ml/100 g and a V0.1 μm/V0.2 μm ratio of greater than or equal to 2.5.Preferably, they are obtained from dehydrated hydrargillite.","Preferably, they are in the form of beads or of extruded materials, or of crushed materials, or in the form of monoliths.","The subjects of the invention are also a catalyst support, especially for the petroleum or petrochemical industry, consisting of alumina agglomerates of the abovementioned type, an intrinsic catalyst, especially for the petroleum or petrochemical industry, consisting of alumina agglomerates of the abovementioned type, and an adsorbent, especially for the petroleum or petrochemical industry, consisting of alumina agglomerates of the abovementioned type.","The subject of the invention is also a process for producing alumina agglomerates of the abovementioned type, in which: an aluminum hydroxide or oxyhydroxide, preferably hydrargillite, undergoes flash dehydration in order to obtain an active alumina powder; said active alumina powder undergoes a forming operation so as to obtain beads with a green fill density of between 500 and 1100 kg/m3, preferably between 700 and 950 kg/m3 inclusive, and a diameter predominantly between 0.8 and 10 mm, preferably between 1 and 5 mm; said beads undergo a heat treatment so as to provide them with a specific surface area of between 50 and 420 m2/g; said beads undergo a hydrothermal treatment by impregnation with water or an aqueous solution, preferably aqueous acid solution, followed by residence in an autoclave at a temperature of above 80° C.; and the agglomerates thus obtained are calcined, preferably between 500 and 1300° C. After the heat treatment that follows granulation, the alumina beads preferably have a V37 Å of greater than 65 ml/100 g, preferably greater than 70 ml/100 g. The active alumina powder obtained after dehydrating the aluminum hydroxide or oxyhydroxide is preferably ground in order to obtain a powder with a median particle size d50 of preferably between 5 and 25 μm.","The active alumina powder obtained after dehydrating the aluminum hydroxide or oxyhydroxide is washed, preferably with water or an aqueous acid solution.","Before granulation of said active alumina powder, the latter preferably undergoes a flash operation.","The subject of the invention is also a process for producing alumina agglomerates of the abovementioned type, in which: an alumina-based material is mixed and extruded in order to form it; the extruded materials thus obtained are subjected to a heat treatment so as to give them a specific surface area of between 50 and 420 m2/g; said extruded materials are subjected to a hydrothermal treatment by impregnation with water or with an aqueous solution, preferably an aqueous acid solution, followed by residence in an autoclave at a temperature of above 80° C.; and the agglomerates thus obtained are calcined, preferably between 500 and 1300° C. Said alumina-based material is preferably dehydrated hydrargillite.","Said alumina-based material may also come from the precipitation of boehmite, pseudo-boehmite or bayerite, or a mixture of such materials.","During the forming of said alumina powder or of said alumina-based material in one of the above processes, one or more pore-forming materials that disappear on heating are preferably added to it.","Said pore-forming materials are preferably chosen from wood flour, charcoal, sulfur, tars, plastics or emulsions of plastics, polyvinyl alcohols, naphthalene.","Said hydrothermal treatment is preferably carried out at a temperature of 150 to 270° C., preferably from 170 to 250° C., for a time of greater than 45 minutes, preferably from 1 to 24 hours and even more preferably from 1.5 to 12 hours.","Said hydrothermal treatment is preferably carried out using an aqueous acid solution containing one or more mineral and/or organic acids.","Said aqueous acid solution also preferably includes one or more compounds that can release anions capable of combining with the aluminum ions.","As will have been understood, the invention consists in obtaining, in the case of alumina agglomerates, a particular distribution of the volumes occupied by the pores of various diameter classes.","It is under these conditions that mechanical strength properties equal to, or even better than, those of the usual agglomerates having an equivalent or lower overall porosity are obtained.","This particular distribution may be obtained using an agglomerate manufacturing process that is distinguished from the usual processes by the execution of the alumina forming step according to the particular methods of implementation that will lead to agglomerates having the desired characteristics finally being obtained.","This forming operation is followed by a short high-temperature heat treatment, or even also preceded by such a treatment.","The invention will be more clearly understood on reading the description that follows, given with reference to the single appended FIGURE.","The latter shows the pore distribution of an alumina A obtained as intermediate product in an example of implementation of the process according to the invention and the pore distribution of a final alumina B according to the invention, obtained from this alumina A.","In the rest of the description, the various parameters that will be referred to are defined below.","The morphology of the aluminas is defined by the indicator of the volumes occupied by the pores having a diameter of greater than or equal to a series of given diameters, namely: the volume occupied by the pores having a diameter greater than or equal to 37 Å (V37 Å); the volume occupied by the pores having a diameter of greater than or equal to 0.1 μm (V0.1 μm); the volume occupied by the pores having a diameter of greater than or equal to 0.2 μm (V0.2 μm); and the volume occupied by the pores having a diameter of greater than or equal to 1 μm (V1 μm).","These volumes may be measured conventionally by the technique called “mercury porosimetry”.","For this purpose, the alumina specimen is placed in a column into which mercury at a pressure P is introduced.","Since mercury does not wet the alumina, its penetration or its non-penetration into the pores of the specimen having a given diameter depends on the value of P. The finest pores require, in order to be filled, a higher pressure P to be established than that for filling the coarser pores.","By measuring the amount of mercury penetrating the specimen for various values of P it is possible to determine the volume occupied by the pores of a diameter greater than the given values of this diameter.","The mechanical strength of the specimen is measured by two parameters: the measured particle-to-particle crush strength (Spp) and the measured value in the shear test.","To measure the Spp of agglomerates, said agglomerates are initially pretreated for two hours at 300° C. in an oven and then left to cool in a dessicator.","They are then taken one by one and placed between the hammer and the anvil of a compressometer until fracture.","The final result is expressed in the form of a mean fracture force measured on a population of about twenty agglomerates.","The objective of the shear test is to determine the mechanical resistance to friction of spherical agglomerates.","The specimen is placed in a ring-shaped cell and pressure is applied to the alumina by means of a cylinder having the same shape as the cell, which is then rotated.","The specimen thus undergoes shear forces leading to a reduction in the size of the beads because of the rotation of the cell under the pressure exerted, the base of the cylinder and the bottom of the cell being also notched.","The analysis reported here was carried out under a pressure of 0.4 bar, the shear being 50.The results are expressed as the percentage undersize for a screen with 1.7 mm square meshes.","A process for manufacturing alumina agglomerates according to the invention will now be described.","The process starts, as in previously known processes, with intense “flash” heating resulting in sudden dehydration of an aluminum hydroxide (hydrargillite, gibbsite or bayerite) or of an aluminum oxyhydroxide (boehmite or diaspore) by means of a stream of hot gas for removing and carrying away the evaporated water very rapidly.","The temperature is around 400 to 1200° C. and the time during which the material to be dehydrated is in contact with the gases is from the order of a fraction of a second to four or five seconds.","Active alumina in powder form, having a median particle size d50 of about 40 μm, is thus obtained.","As starting compound, it is preferred to use hydrargillite.","Experiments have shown that this compound is the most favorable for obtaining a final product having the desired properties.","In addition, it is relatively inexpensive.","The powder obtained then undergoes, preferably and as is known, a grinding operation that brings its median particle size d50 to about 15 μm, generally between 5 and 25 μm.","Next, as is known, the powder is washed with water or with acidified water, this having in particular the purpose of reducing its alkali metal content.","According to the invention, the powder then preferably undergoes a flash operation under conditions similar to those of the flash operation described above so as to continue to develop its porosity.","Next, according to the invention, an operation of forming the alumina is carried out.","According to the preferred method of implementing the invention, the alumina powder is granulated by means of a rotational technique such as, for example, a rotary pelletizer or rotary drum.","This type of process makes it possible to obtain beads of controlled pore diameters and distribution, these dimensions and distributions generally being created during the agglomeration step.","The porosity may be created by various means, such as the choice of particle size of the alumina powder or the agglomeration of several alumina powders of different particle sizes.","Another method consists in mixing with the alumina powder, before or during the agglomeration step, one or more compounds, called pore formers, that disappear when heated and thus create porosity in the beads.","As pore-forming compounds used, mention may be made, by way of example, of wood flour, charcoal, sulfur, tars, plastics or emulsions of plastics, such as polyvinyl chloride and polyvinyl alcohol, naphthalene or the like.","The amount of pore-forming compounds added is determined by the desired volume.","The aim is to produce beads with a green fill density of between 500 and 1100 kg/m3, preferably between 700 and 950 kg/m3, for example 810 kg/m3, and a diameter of predominantly between 0.8 and 10 mm, preferably between 1 and 5 mm.","This granulation is itself a conventional process, but the addition of the invention lies essentially in the choice of the characteristics of the beads resulting from the granulation.","These characteristics, in combination with the subsequent treatments undergone by the beads, will determine the properties of the agglomerates obtained at the end of the treatment.","Next, again according to the invention, the beads undergo a heat treatment that allows an alumina to be obtained whose specific surface area is from 50 to 420 m2/g.","Below 50 m2/g, the reactivity of the alumina during the subsequent autoclaving operation would not be sufficient, and a specific surface area of greater than 420 m2/g would correspond to pores in the final product that are too small for the applications envisioned.","This alumina will be called hereafter “alumina A”.","An example of the pore distribution of such an alumina A is shown in the single figure.","In this example, the specific surface area of this alumina A is 187 m2/g.","In this single FIGURE, the pore diameter (in μm) is plotted on the x-axis and the cumulative pore volume (in ml/100 g), that is to say the volume occupied by the pores having diameters greater than or equal to the diameter on the x-axis, is plotted on the y-axis.","To obtain a final alumina according to the invention, it is recommended to give alumina A a V37 Å of greater than 65 ml/100 g, usually greater than 70 ml/100 g. This alumina A then undergoes a hydrothermal treatment similar to those known in the prior art, for example in the aforementioned document EP-A-0 387 109.To give an example, this treatment may consist of impregnation of alumina A with an aqueous solution containing 4% aluminum nitrate and 9% formic acid (these percentages being calculated on a weight basis with respect to the weight of alumina introduced), followed by residence of the impregnated alumina A in an autoclave with a rotating basket at 200° C. for 5 h 30 min.","Generally speaking, the hydrothermal treatment is carried out at a temperature of preferably between 150 and 270° C., advantageously between 170 and 250° C. inclusive.","The duration of said treatment is in general greater than 45 minutes, preferably between 1 and 24 hours and even more advantageously between 1.5 and 12 hours inclusive.","The aqueous acid impregnation solution comprises one or more mineral and/or organic acids.","To give an example, mention may be made of nitric acid, hydrochloric acid, perchloric acid, sulfuric acid and weak acids, the solution of which has a pH of less than 4, such as acetic acid or formic acid.","It may also contain one or more compounds that can release anions capable of combining with the aluminum ions.","Thus, by way of example, mention may be made of compounds comprising a nitrate (such as aluminum nitrate), chloride, sulfate, perchlorate, chloroacetate, trichloroacetate, bromo-acetate or dibromoacetate ion, and the anions of general formula: R—COO− such as formates and acetates.","Finally, as is usual, the alumina undergoes a final heat treatment (or calcination) at high temperature (typically between 400 and 1300° C., for example at 800° C.), which allows the desired specific surface area to be obtained.","Beads are the preferred form of the invention, but it is also conceivable to use, for example, materials in extruded form or crushed materials or monoliths, and also to use alumina in powder form.","According to an alternative method of implementing the invention, the alumina may also be in the form of alumina extrudates.","These are generally obtained by mixing and then extruding an alumina-based material followed by calcination and then, in succession, as described above, impregnation, hydrothermal treatment and then post-forming heat treatment.","The starting material may vary very widely in nature: it may result from the partial and rapid dehydration of hydrargillite, or from the precipitation of boehmite, pseudo-boehmite, bayerite or of a mixture of these aluminas.","During mixing, the alumina may be mixed with additives, especially pore formers such as those defined above.","The single FIGURE shows the pore distribution of an alumina according to the invention, called “alumina B”, produced according to the particular example of hydrothermal and calcination treatment that was described in the case of the above alumina A.","The calcination gave this alumina B a specific surface area of 110 m2/g.","The single FIGURE shows that the hydrothermal and calcination treatment operations resulted in an alumina B being obtained in which, compared with the starting alumina A, the volume occupied by the pores of small size (<0.04 μm) has substantially increased.","Above all, the volume occupied by the pores having a size greater than 0.2 μm decreased considerably, to the point of becoming almost negligible.","The aluminas according to the invention are preferably obtained from hydrargillite; they have a V37 Å of at least 75 ml/100 g, preferably greater than 80 ml/100 g and even more preferably greater than 85 ml/100 g. At the same time, their V0.1 μm is less than or equal to 30 ml/100 g, preferably less than or equal to 25 ml/100 g and even more preferably less than 20 ml/100 g, or even less than 15 ml/100 g. They have a V0.2 μm of less than or equal to 20 ml/100 g, advantageously less than or equal to 15 ml/100 g or even less than or equal to 10 ml/100 g. As regards the V1 μm, it is beneficial for it not to exceed 7 ml/100 g, or even 5.5 ml/100 g, preferably 4 ml/100 g. The V0.1 μm/V0.2 μm ratio is preferably greater than or equal to 1.5, or even greater than or equal to 2, and even more advantageously greater than or equal to 2.5.Under these conditions, and when the aluminas according to the invention are in the form of beads having diameters of between 2.0 and 2.8 mm, they have a minimum Spp of 2 daN, preferably equal to or greater than 2.5 daN, or even equal to or greater than 3 daN, and even more advantageously exceeding 3.5 daN.","As regards the shear test measured under the abovementioned conditions, this gives values of less than 4%, usefully less than 3% and even more advantageously less than 2% or even less than 1%.","Table I gives the characteristics of several aluminas according to the invention, obtained by varying the experimental conditions that have determined the forming of the beads and the following hydrothermal treatment.","Their particle size is in this case systematically between 2.0 and 2.8 mm inclusive.","A comparison is made with aluminas produced in a similar manner, with the same particle size, but without taking the same care of controlling the porosity profile.","All the specific surface areas were normalized, by calcination, to 105±5 m2/g in order to allow a comparison to be made that excludes the influence of this parameter.","Aluminas 1 to 8 are according to the invention, while aluminas 9 to 11 are comparative examples not according to the invention.","The specific surface areas are given in m2/g, all the volumes are in ml/100 g, the Spp values are in daN and the measurements obtained from the shear test are in % (cf.","operating methods above).","TABLE I Characteristics of the aluminas described Specific surface Alumina area V37 Å V0.1 μm V0.2 μm V1 μm V0.1 μm/V0.2 μm Spp Shear I 1 110 93.9 19.4 6.6 2.7 2.9 3.4 0.8 n 2 104 92.2 30.3 9.4 3.2 3.2 3.0 0.5 v 3 105 84.3 14.0 6.9 2.1 2.0 4.5 0.7 e 4 103 87.2 27.8 13.5 3.8 2.1 3.3 1.5 n 5 106 87.2 26.4 17.5 5.3 1.5 3.0 1.6 t 6 105 101.2 30.5 15.5 5.0 2.0 2.9 1.8 i 7 110 86.2 7.0 2.7 2.1 2.6 6.6 0.5 o 8 102 87.8 9.9 3.8 2.0 2.6 6.3 0.9 n Comparative 9 105 91.6 34.0 24.9 7.4 1.4 1.6 6.9 examples 10 100 87.2 31.3 21.6 7.1 1.4 2.0 4.3 11 104 85.7 32.3 22.5 5.9 1.4 1.8 4.9 As may be seen, precise control of the density of the pores of diameter greater than or equal to 0.1 μm and of the pores having diameters of greater than or equal to 0.2 4m is, unexpectedly and in a surprising proportion, given the prior knowledge, essential for achieving agglomerates with the maximum mechanical strength independently of the level of total porosity of the alumina.","All the specimens according to the invention have a V37 Å of greater than 75 ml/100 g and a V0.1 μm of less than 31 ml/100 g. They also have a V0.2 μm of less than 20 ml/100 g. They also have a V1 μm of less than 7 ml/100 g and a V0.1 μm/V0.2 μm ratio of at least 1.5, in accordance with the preferred characteristics of the invention.","However, it should be pointed out that the features that have been given as preferred are independent of one another and that it would remain in accordance with the invention to comply with some of them without complying with the others.","It should be noted that the control specimens 9 to 11 all have a high V37 Å, in accordance with one of the obligatory characteristics of the aluminas of the invention.","However, they have a V0.1 μm and a V0.2 μm that are higher than the maximum required by the invention, coupled with a V1 μm of greater than the preferred upper limit of the invention.","This means that these specimens have a quantity of large-diameter pores that is relatively higher than that required by the invention.","However, these specimens have only relatively mediocre mechanical properties as regards crush strength (Spp) and resistance to friction (shear).","This major influence of the presence of large-diameter pores is also found when the results of specimens 1 to 8 according to the invention are examined.","The best mechanical strength results are obtained for specimens 7 and 8, the V37 Å of which is not exceptionally high, and these specimens are distinguished from the others by particularly low V0.1 μm, V0.2 μm and V1 μm values.","It has been found that this pore distribution gives these specimens a crush strength very substantially higher than that of the other specimens, although their shear strength is at least very good.","The best results are obtained with specimen 7 that has the lowest V0.1 μm and V0.2 μm values of the series.","It may also be noted that specimens (1, 2, 6) having a very high V37 Å, and therefore a very high overall porosity, but V0.1 μm, V0.2 μm, V1 μm and V0.1 μm/V0.2 μm values within the necessary or preferred limits of the invention, nevertheless have mechanical properties that are still substantially better than those of the control specimens.","However, these have a lower overall porosity that would a priori lead to a better mechanical strength.","Here again, controlling the presence of pores having diameters of greater than 0.1 μm, or even 0.2 μm and 1 μm, under the specified conditions appears to be particularly advantageous.","Measuring the V0.1 μm/V0.2 μm ratio gives an indicator of what the distribution of the pore diameters in the 0.1-0.2 μm range should be.","The invention is applicable for obtaining catalyst supports in the form of beads, especially those for the petroleum industry and the petrochemical industry, in which the combination of a high porosity and a high mechanical strength of the supports gives effective catalysts with a long lifetime.","These catalysts may be used, for example, in hydrotreatment, particularly in hydrometallation, to reduce the metal content of a hydrocarbon cut.","These alumina agglomerates can also be used as intrinsic catalysts or as adsorbents."]],"string":"[\n [\n \"Alumina agglomerates, the preparation method thereof and use of same as an absorbent or catalyst carrier\",\n \"The invention relates to alumina agglomerates of the type obtained by dehydrating an aluminium oxyhydroxide or hydroxide, agglomerating the alumina thus obtained, hydrothermally treating the agglomerates and calcinating same.\",\n \"Said agglomerates are characterised in that: the V37 Å thereof is greater than or equal to 75 ml/100 g, preferably greater than or equal to 80 ml/100 g and, better still, greater than or equal to 85 ml/100 g; the V0.1 μm thereof is less than or equal to 31 ml/100 g; and the V0.2 μm thereof is less than or equal to 20 ml/100 g, preferably less than or equal to 15 ml/100 g and, better still, less than or equal to 10 ml/100 g. The invention also relates to a catalyst carrier, an intrinsic catalyst or an absorbent, in particular for use in the petroleum and petrochemical industry, comprising such alumina agglomerates.\",\n \"Moreover, the invention relates to methods for preparing said agglomerates.\"\n ],\n [\n \"1.Alumina agglomerates of the type obtained by the treatment of an aluminum hydroxide or oxyhydroxide, agglomeration of the alumina thus obtained, hydrothermal treatment of the agglomerates and calcination, wherein they have a V37 Å of greater than or equal to 75 ml/100 g, preferably greater than or equal to 80 ml/100 g and even more preferably greater than or equal to 85 ml/100 g; they have a V0.1 μm of less than or equal to 31 ml/l00 g, preferably less than or equal to 25 ml/100 g, even more preferably less than or equal to 20 ml/100 g and optimally less than or equal to 15 ml/100 g; and wherein: they have a V0.2 μm of less than or equal to 20 ml/100 g, preferably less than or equal to 15 ml/100 g and even more preferably less than or equal to 10 ml/100 g. 2.The alumina agglomerates as claimed in claim 1, wherein they have a V1 μm of less than or equal to 7 ml/100 g, preferably less than or equal to 5.5 ml/100 g and even more preferably less than or equal to 4 ml/100 g. 3.The alumina agglomerates as claimed in claim 1 wherein they have a V0.1 μm/V0.2 μm ratio of greater than or equal to 1.5, preferably greater than or equal to 2 and even more preferably greater than or equal to 2.5.4.The alumina agglomerates as claimed in claim 1 wherein they have simultaneously a V37 Å of greater than or equal to 80 ml/100 g, a V0.1 μm of less than or equal to 15 ml/100 g, a V0.2 μm of less than or equal to 10 ml/100 g, a V1 μm of less than or equal to 4 ml/100 g and a V0.1 μm/V0.2 μm ratio of greater than or equal to 2.5.5.The alumina agglomerates as claimed in claim 1 wherein they have been obtained from dehydrated hydrargillite.\",\n \"6.The alumina agglomerates as claimed in claim 1 wherein they are in the form of beads.\",\n \"7.The alumina agglomerates as claimed in claim 1 wherein they are in the form of extruded materials.\",\n \"8.The alumina agglomerates as claimed in claim 1 wherein they are in the form of crushed materials.\",\n \"9.The alumina agglomerates as claimed in claim 1 wherein they are in the form of monoliths.\",\n \"10.A catalyst support, especially for the petroleum of petrochemical industry, wherein it consists of alumina agglomerates as claimed in claim 1.11.An intrinsic catalyst, especially for the petroleum or petrochemical industry, wherein it consists of alumina agglomerates as claimed in claim 1.12.An adsorbent, especially for the petroleum or petrochemical industry, wherein it consists of alumina agglomerates as claimed in claim 1.13.A process for producing alumina agglomerates as claimed in claim 6, in which: an aluminum hydroxide or oxyhydroxide, preferably hydrargillite, undergoes flash dehydration in order to obtain an active alumina powder; said active alumina powder undergoes a forming operation so as to obtain beads with a green fill density of between 500 and 1100 kg/m3, preferably between 700 and 950 kg/m3 inclusive, and a diameter predominantly between 0.8 and 10 mm, preferably between 1 and 5 mm; said beads undergo a heat treatment so as to provide them with a specific surface area of between 50 and 420 m2/g; said beads undergo a hydrothermal treatment by impregnation with water or an aqueous solution, preferably aqueous acid solution, followed by residence in an autoclave at a temperature of above 80° C.; and the agglomerates thus obtained are calcined, preferably between 500 and 1300° C. 14.The process as claimed in claim 13, wherein after the heat treatment that follows granulation, the alumina beads preferably have a V37 Å of greater than 65 ml/100 g, preferably greater than 70 ml/100 g. 15.The process as claimed in claim 13 wherein the active alumina powder obtained after dehydrating the aluminum hydroxide or oxyhydroxide is ground in order to obtain a powder with a median particle size d50 of preferably between 5 and 25 μm.\",\n \"16.The process as claimed in claim 13 wherein the active alumina powder obtained after dehydrating the aluminum hydroxide or oxyhydroxide is washed with water or an aqueous acid solution.\",\n \"17.The process as claimed in claim 13 wherein prior to granulation of said active alumina powder, the latter undergoes a flash operation.\",\n \"18.The process for producing alumina agglomerates as claimed in claim 7, in which: an alumina-based material is mixed and extruded in order to form it; the extruded materials thus obtained are subjected to a heat treatment so as to give them a specific surface area of between 50 and 420 m2/g; said extruded materials are subjected to a hydrothermal treatment by impregnation with water or with an aqueous solution, preferably an aqueous acid solution, followed by residence in an autoclave at a temperature of above 80° C.; and the agglomerates thus obtained are calcined, preferably between 500 and 1300° C. 19.The process as claimed in claim 18, wherein said alumina-based material is dehydrated hydrargillite.\",\n \"20.The process as claimed in claim 18, wherein alumina-based material comes from the precipitation of boehmite, pseudo-boehmite or bayerite, or a mixture of such materials.\",\n \"21.The process as claimed in claim 13 wherein during the forming of said alumina powder or of said alumina-based material, one or more pore-forming materials that disappear on heating are added to it.\",\n \"22.The process as claimed in claim 21, wherein said pore-forming materials are chosen from wood flour, charcoal, sulfur, tars, plastics or emulsions of plastics, polyvinyl alcohols, naphthalene.\",\n \"23.The process as claimed in claim 13 wherein said hydrothermal treatment is carried out at a temperature of 150 to 270° C., preferably from 170 to 250° C., for a time of greater than 45 minutes, preferably from 1 to 24 hours and even more preferably from 1.5 to 12 hours.\",\n \"24.The process as claimed in claim 13 wherein said hydrothermal treatment is carried out using an aqueous acid solution containing one or more mineral and/or organic acids.\",\n \"25.The process as claimed in claim 24, wherein said aqueous acid solution also includes one or more compounds that can release anions capable of combining with the aluminum ions.\"\n ],\n [\n \"The invention relates to the field of alumina agglomerates that can be used especially as adsorbents, as catalyst supports or as catalysts.\",\n \"More precisely, it relates to novel alumina agglomerates having specific physical characteristics, in particular from the standpoint of their porosity, giving them particularly advantageous mechanical properties.\",\n \"The invention also relates to a process for obtaining such agglomerates.\",\n \"Finally, it relates to their use as catalyst supports, as catalysts or as adsorbents.\",\n \"Certain catalytic processes, such as heterogeneous catalysis, for example in the field of treatment of petroleum oil cuts or the treatment of gaseous effluents, such as the exhaust gases from internal combustion engines, require the use of supports having a high porosity and good mechanical properties, such as good crush resistance and/or attrition resistance.\",\n \"Alumina-based supports meet these criteria, especially when they have undergone a hydrothermal treatment, for example after the alumina has been made in the form of agglomerates.\",\n \"This is because such a treatment substantially improves the mechanical properties of the agglomerates.\",\n \"This hydrothermal treatment consists in impregnating the alumina agglomerates with water or an aqueous acid solution, and then heating these agglomerates, placed inside an autoclave, to a temperature of above 80° C. European Patent No.\",\n \"EP-A-0 387 109, in the name of the Applicant, describes an example of an advantageous process for carrying out this hydrothermal treatment.\",\n \"The agglomerates thus treated are then dried and then subjected to a heat treatment called “calcination” at a given temperature.\",\n \"This calcination temperature is chosen according to the desired specific surface area and to the temperature at which these supports will be used.\",\n \"The efficiency of such alumina agglomerates is to a large part dependent on their porosity: a high porosity is often favorable for the various applications envisioned, because of the lower diffusional and/or tortuosity constraints.\",\n \"In applications in which the support will be used not only for satisfactorily dispersing a metallic active phase or phases supplied during production of the catalyst, but also to retain contaminants, such as during a hydrometallation operation, a high porosity is advantageous.\",\n \"However, a high porosity is generally regarded as going hand in hand with a degradation of the mechanical strength of the material.\",\n \"The use of alumina agglomerates with a high porosity is therefore difficult in important applications, specially in catalyst supports for which the mechanical strength of the alumina to a large extent governs the duration of use of the catalyst.\",\n \"It turns out that degradation of the mechanical strength of the catalyst during use has dramatic effects, both technically and economically, on the operating process.\",\n \"The object of the invention is to provide alumina agglomerates users with materials, exhibiting an excellent compromise between high porosity and mechanical strength that makes them compatible with demanding applications from the latter standpoint.\",\n \"For this purpose, the subject of the invention is alumina agglomerates of the type obtained by the treatment of an aluminum hydroxide or oxyhydroxide, agglomeration of the alumina thus obtained, hydrothermal treatment of the agglomerates and calcination, characterized in that: they have a V37 Å of greater than or equal to 75 ml/100 g, preferably greater than or equal to 80 ml/100 g and even more preferably greater than or equal to 85 ml/100 g; they have a V0.1 μm of less than or equal to 31 ml/100 g, preferably less than or equal to 25 ml/100 g, even more preferably less than or equal to 20 ml/100 g and optimally less than or equal to 15 ml/100 g; and in that: they have a V0.2 μm of less than or equal to 20 ml/100 g, preferably less than or equal to 15 ml/100 g and even more preferably less than or equal to 10 ml/100 g. Preferably, they have a V1 μm of less than or equal to 7 ml/100 g, preferably less than or equal to 5.5 ml/100 g and even more preferably less than or equal to 4 ml/100 g. Preferably, they have a V0.1 μm/V0.2 μm ratio of greater than or equal to 1.5, preferably greater than or equal to 2 and even more preferably greater than or equal to 2.5.Preferably, they have simultaneously a V37 Å of greater than or equal to 80 ml/100 g, a V0.1 μm of less than or equal to 15 ml/100 g, a V0.2 μm of less than or equal to 10 ml/100 g, a V1 μm of less than or equal to 4 ml/100 g and a V0.1 μm/V0.2 μm ratio of greater than or equal to 2.5.Preferably, they are obtained from dehydrated hydrargillite.\",\n \"Preferably, they are in the form of beads or of extruded materials, or of crushed materials, or in the form of monoliths.\",\n \"The subjects of the invention are also a catalyst support, especially for the petroleum or petrochemical industry, consisting of alumina agglomerates of the abovementioned type, an intrinsic catalyst, especially for the petroleum or petrochemical industry, consisting of alumina agglomerates of the abovementioned type, and an adsorbent, especially for the petroleum or petrochemical industry, consisting of alumina agglomerates of the abovementioned type.\",\n \"The subject of the invention is also a process for producing alumina agglomerates of the abovementioned type, in which: an aluminum hydroxide or oxyhydroxide, preferably hydrargillite, undergoes flash dehydration in order to obtain an active alumina powder; said active alumina powder undergoes a forming operation so as to obtain beads with a green fill density of between 500 and 1100 kg/m3, preferably between 700 and 950 kg/m3 inclusive, and a diameter predominantly between 0.8 and 10 mm, preferably between 1 and 5 mm; said beads undergo a heat treatment so as to provide them with a specific surface area of between 50 and 420 m2/g; said beads undergo a hydrothermal treatment by impregnation with water or an aqueous solution, preferably aqueous acid solution, followed by residence in an autoclave at a temperature of above 80° C.; and the agglomerates thus obtained are calcined, preferably between 500 and 1300° C. After the heat treatment that follows granulation, the alumina beads preferably have a V37 Å of greater than 65 ml/100 g, preferably greater than 70 ml/100 g. The active alumina powder obtained after dehydrating the aluminum hydroxide or oxyhydroxide is preferably ground in order to obtain a powder with a median particle size d50 of preferably between 5 and 25 μm.\",\n \"The active alumina powder obtained after dehydrating the aluminum hydroxide or oxyhydroxide is washed, preferably with water or an aqueous acid solution.\",\n \"Before granulation of said active alumina powder, the latter preferably undergoes a flash operation.\",\n \"The subject of the invention is also a process for producing alumina agglomerates of the abovementioned type, in which: an alumina-based material is mixed and extruded in order to form it; the extruded materials thus obtained are subjected to a heat treatment so as to give them a specific surface area of between 50 and 420 m2/g; said extruded materials are subjected to a hydrothermal treatment by impregnation with water or with an aqueous solution, preferably an aqueous acid solution, followed by residence in an autoclave at a temperature of above 80° C.; and the agglomerates thus obtained are calcined, preferably between 500 and 1300° C. Said alumina-based material is preferably dehydrated hydrargillite.\",\n \"Said alumina-based material may also come from the precipitation of boehmite, pseudo-boehmite or bayerite, or a mixture of such materials.\",\n \"During the forming of said alumina powder or of said alumina-based material in one of the above processes, one or more pore-forming materials that disappear on heating are preferably added to it.\",\n \"Said pore-forming materials are preferably chosen from wood flour, charcoal, sulfur, tars, plastics or emulsions of plastics, polyvinyl alcohols, naphthalene.\",\n \"Said hydrothermal treatment is preferably carried out at a temperature of 150 to 270° C., preferably from 170 to 250° C., for a time of greater than 45 minutes, preferably from 1 to 24 hours and even more preferably from 1.5 to 12 hours.\",\n \"Said hydrothermal treatment is preferably carried out using an aqueous acid solution containing one or more mineral and/or organic acids.\",\n \"Said aqueous acid solution also preferably includes one or more compounds that can release anions capable of combining with the aluminum ions.\",\n \"As will have been understood, the invention consists in obtaining, in the case of alumina agglomerates, a particular distribution of the volumes occupied by the pores of various diameter classes.\",\n \"It is under these conditions that mechanical strength properties equal to, or even better than, those of the usual agglomerates having an equivalent or lower overall porosity are obtained.\",\n \"This particular distribution may be obtained using an agglomerate manufacturing process that is distinguished from the usual processes by the execution of the alumina forming step according to the particular methods of implementation that will lead to agglomerates having the desired characteristics finally being obtained.\",\n \"This forming operation is followed by a short high-temperature heat treatment, or even also preceded by such a treatment.\",\n \"The invention will be more clearly understood on reading the description that follows, given with reference to the single appended FIGURE.\",\n \"The latter shows the pore distribution of an alumina A obtained as intermediate product in an example of implementation of the process according to the invention and the pore distribution of a final alumina B according to the invention, obtained from this alumina A.\",\n \"In the rest of the description, the various parameters that will be referred to are defined below.\",\n \"The morphology of the aluminas is defined by the indicator of the volumes occupied by the pores having a diameter of greater than or equal to a series of given diameters, namely: the volume occupied by the pores having a diameter greater than or equal to 37 Å (V37 Å); the volume occupied by the pores having a diameter of greater than or equal to 0.1 μm (V0.1 μm); the volume occupied by the pores having a diameter of greater than or equal to 0.2 μm (V0.2 μm); and the volume occupied by the pores having a diameter of greater than or equal to 1 μm (V1 μm).\",\n \"These volumes may be measured conventionally by the technique called “mercury porosimetry”.\",\n \"For this purpose, the alumina specimen is placed in a column into which mercury at a pressure P is introduced.\",\n \"Since mercury does not wet the alumina, its penetration or its non-penetration into the pores of the specimen having a given diameter depends on the value of P. The finest pores require, in order to be filled, a higher pressure P to be established than that for filling the coarser pores.\",\n \"By measuring the amount of mercury penetrating the specimen for various values of P it is possible to determine the volume occupied by the pores of a diameter greater than the given values of this diameter.\",\n \"The mechanical strength of the specimen is measured by two parameters: the measured particle-to-particle crush strength (Spp) and the measured value in the shear test.\",\n \"To measure the Spp of agglomerates, said agglomerates are initially pretreated for two hours at 300° C. in an oven and then left to cool in a dessicator.\",\n \"They are then taken one by one and placed between the hammer and the anvil of a compressometer until fracture.\",\n \"The final result is expressed in the form of a mean fracture force measured on a population of about twenty agglomerates.\",\n \"The objective of the shear test is to determine the mechanical resistance to friction of spherical agglomerates.\",\n \"The specimen is placed in a ring-shaped cell and pressure is applied to the alumina by means of a cylinder having the same shape as the cell, which is then rotated.\",\n \"The specimen thus undergoes shear forces leading to a reduction in the size of the beads because of the rotation of the cell under the pressure exerted, the base of the cylinder and the bottom of the cell being also notched.\",\n \"The analysis reported here was carried out under a pressure of 0.4 bar, the shear being 50.The results are expressed as the percentage undersize for a screen with 1.7 mm square meshes.\",\n \"A process for manufacturing alumina agglomerates according to the invention will now be described.\",\n \"The process starts, as in previously known processes, with intense “flash” heating resulting in sudden dehydration of an aluminum hydroxide (hydrargillite, gibbsite or bayerite) or of an aluminum oxyhydroxide (boehmite or diaspore) by means of a stream of hot gas for removing and carrying away the evaporated water very rapidly.\",\n \"The temperature is around 400 to 1200° C. and the time during which the material to be dehydrated is in contact with the gases is from the order of a fraction of a second to four or five seconds.\",\n \"Active alumina in powder form, having a median particle size d50 of about 40 μm, is thus obtained.\",\n \"As starting compound, it is preferred to use hydrargillite.\",\n \"Experiments have shown that this compound is the most favorable for obtaining a final product having the desired properties.\",\n \"In addition, it is relatively inexpensive.\",\n \"The powder obtained then undergoes, preferably and as is known, a grinding operation that brings its median particle size d50 to about 15 μm, generally between 5 and 25 μm.\",\n \"Next, as is known, the powder is washed with water or with acidified water, this having in particular the purpose of reducing its alkali metal content.\",\n \"According to the invention, the powder then preferably undergoes a flash operation under conditions similar to those of the flash operation described above so as to continue to develop its porosity.\",\n \"Next, according to the invention, an operation of forming the alumina is carried out.\",\n \"According to the preferred method of implementing the invention, the alumina powder is granulated by means of a rotational technique such as, for example, a rotary pelletizer or rotary drum.\",\n \"This type of process makes it possible to obtain beads of controlled pore diameters and distribution, these dimensions and distributions generally being created during the agglomeration step.\",\n \"The porosity may be created by various means, such as the choice of particle size of the alumina powder or the agglomeration of several alumina powders of different particle sizes.\",\n \"Another method consists in mixing with the alumina powder, before or during the agglomeration step, one or more compounds, called pore formers, that disappear when heated and thus create porosity in the beads.\",\n \"As pore-forming compounds used, mention may be made, by way of example, of wood flour, charcoal, sulfur, tars, plastics or emulsions of plastics, such as polyvinyl chloride and polyvinyl alcohol, naphthalene or the like.\",\n \"The amount of pore-forming compounds added is determined by the desired volume.\",\n \"The aim is to produce beads with a green fill density of between 500 and 1100 kg/m3, preferably between 700 and 950 kg/m3, for example 810 kg/m3, and a diameter of predominantly between 0.8 and 10 mm, preferably between 1 and 5 mm.\",\n \"This granulation is itself a conventional process, but the addition of the invention lies essentially in the choice of the characteristics of the beads resulting from the granulation.\",\n \"These characteristics, in combination with the subsequent treatments undergone by the beads, will determine the properties of the agglomerates obtained at the end of the treatment.\",\n \"Next, again according to the invention, the beads undergo a heat treatment that allows an alumina to be obtained whose specific surface area is from 50 to 420 m2/g.\",\n \"Below 50 m2/g, the reactivity of the alumina during the subsequent autoclaving operation would not be sufficient, and a specific surface area of greater than 420 m2/g would correspond to pores in the final product that are too small for the applications envisioned.\",\n \"This alumina will be called hereafter “alumina A”.\",\n \"An example of the pore distribution of such an alumina A is shown in the single figure.\",\n \"In this example, the specific surface area of this alumina A is 187 m2/g.\",\n \"In this single FIGURE, the pore diameter (in μm) is plotted on the x-axis and the cumulative pore volume (in ml/100 g), that is to say the volume occupied by the pores having diameters greater than or equal to the diameter on the x-axis, is plotted on the y-axis.\",\n \"To obtain a final alumina according to the invention, it is recommended to give alumina A a V37 Å of greater than 65 ml/100 g, usually greater than 70 ml/100 g. This alumina A then undergoes a hydrothermal treatment similar to those known in the prior art, for example in the aforementioned document EP-A-0 387 109.To give an example, this treatment may consist of impregnation of alumina A with an aqueous solution containing 4% aluminum nitrate and 9% formic acid (these percentages being calculated on a weight basis with respect to the weight of alumina introduced), followed by residence of the impregnated alumina A in an autoclave with a rotating basket at 200° C. for 5 h 30 min.\",\n \"Generally speaking, the hydrothermal treatment is carried out at a temperature of preferably between 150 and 270° C., advantageously between 170 and 250° C. inclusive.\",\n \"The duration of said treatment is in general greater than 45 minutes, preferably between 1 and 24 hours and even more advantageously between 1.5 and 12 hours inclusive.\",\n \"The aqueous acid impregnation solution comprises one or more mineral and/or organic acids.\",\n \"To give an example, mention may be made of nitric acid, hydrochloric acid, perchloric acid, sulfuric acid and weak acids, the solution of which has a pH of less than 4, such as acetic acid or formic acid.\",\n \"It may also contain one or more compounds that can release anions capable of combining with the aluminum ions.\",\n \"Thus, by way of example, mention may be made of compounds comprising a nitrate (such as aluminum nitrate), chloride, sulfate, perchlorate, chloroacetate, trichloroacetate, bromo-acetate or dibromoacetate ion, and the anions of general formula: R—COO− such as formates and acetates.\",\n \"Finally, as is usual, the alumina undergoes a final heat treatment (or calcination) at high temperature (typically between 400 and 1300° C., for example at 800° C.), which allows the desired specific surface area to be obtained.\",\n \"Beads are the preferred form of the invention, but it is also conceivable to use, for example, materials in extruded form or crushed materials or monoliths, and also to use alumina in powder form.\",\n \"According to an alternative method of implementing the invention, the alumina may also be in the form of alumina extrudates.\",\n \"These are generally obtained by mixing and then extruding an alumina-based material followed by calcination and then, in succession, as described above, impregnation, hydrothermal treatment and then post-forming heat treatment.\",\n \"The starting material may vary very widely in nature: it may result from the partial and rapid dehydration of hydrargillite, or from the precipitation of boehmite, pseudo-boehmite, bayerite or of a mixture of these aluminas.\",\n \"During mixing, the alumina may be mixed with additives, especially pore formers such as those defined above.\",\n \"The single FIGURE shows the pore distribution of an alumina according to the invention, called “alumina B”, produced according to the particular example of hydrothermal and calcination treatment that was described in the case of the above alumina A.\",\n \"The calcination gave this alumina B a specific surface area of 110 m2/g.\",\n \"The single FIGURE shows that the hydrothermal and calcination treatment operations resulted in an alumina B being obtained in which, compared with the starting alumina A, the volume occupied by the pores of small size (<0.04 μm) has substantially increased.\",\n \"Above all, the volume occupied by the pores having a size greater than 0.2 μm decreased considerably, to the point of becoming almost negligible.\",\n \"The aluminas according to the invention are preferably obtained from hydrargillite; they have a V37 Å of at least 75 ml/100 g, preferably greater than 80 ml/100 g and even more preferably greater than 85 ml/100 g. At the same time, their V0.1 μm is less than or equal to 30 ml/100 g, preferably less than or equal to 25 ml/100 g and even more preferably less than 20 ml/100 g, or even less than 15 ml/100 g. They have a V0.2 μm of less than or equal to 20 ml/100 g, advantageously less than or equal to 15 ml/100 g or even less than or equal to 10 ml/100 g. As regards the V1 μm, it is beneficial for it not to exceed 7 ml/100 g, or even 5.5 ml/100 g, preferably 4 ml/100 g. The V0.1 μm/V0.2 μm ratio is preferably greater than or equal to 1.5, or even greater than or equal to 2, and even more advantageously greater than or equal to 2.5.Under these conditions, and when the aluminas according to the invention are in the form of beads having diameters of between 2.0 and 2.8 mm, they have a minimum Spp of 2 daN, preferably equal to or greater than 2.5 daN, or even equal to or greater than 3 daN, and even more advantageously exceeding 3.5 daN.\",\n \"As regards the shear test measured under the abovementioned conditions, this gives values of less than 4%, usefully less than 3% and even more advantageously less than 2% or even less than 1%.\",\n \"Table I gives the characteristics of several aluminas according to the invention, obtained by varying the experimental conditions that have determined the forming of the beads and the following hydrothermal treatment.\",\n \"Their particle size is in this case systematically between 2.0 and 2.8 mm inclusive.\",\n \"A comparison is made with aluminas produced in a similar manner, with the same particle size, but without taking the same care of controlling the porosity profile.\",\n \"All the specific surface areas were normalized, by calcination, to 105±5 m2/g in order to allow a comparison to be made that excludes the influence of this parameter.\",\n \"Aluminas 1 to 8 are according to the invention, while aluminas 9 to 11 are comparative examples not according to the invention.\",\n \"The specific surface areas are given in m2/g, all the volumes are in ml/100 g, the Spp values are in daN and the measurements obtained from the shear test are in % (cf.\",\n \"operating methods above).\",\n \"TABLE I Characteristics of the aluminas described Specific surface Alumina area V37 Å V0.1 μm V0.2 μm V1 μm V0.1 μm/V0.2 μm Spp Shear I 1 110 93.9 19.4 6.6 2.7 2.9 3.4 0.8 n 2 104 92.2 30.3 9.4 3.2 3.2 3.0 0.5 v 3 105 84.3 14.0 6.9 2.1 2.0 4.5 0.7 e 4 103 87.2 27.8 13.5 3.8 2.1 3.3 1.5 n 5 106 87.2 26.4 17.5 5.3 1.5 3.0 1.6 t 6 105 101.2 30.5 15.5 5.0 2.0 2.9 1.8 i 7 110 86.2 7.0 2.7 2.1 2.6 6.6 0.5 o 8 102 87.8 9.9 3.8 2.0 2.6 6.3 0.9 n Comparative 9 105 91.6 34.0 24.9 7.4 1.4 1.6 6.9 examples 10 100 87.2 31.3 21.6 7.1 1.4 2.0 4.3 11 104 85.7 32.3 22.5 5.9 1.4 1.8 4.9 As may be seen, precise control of the density of the pores of diameter greater than or equal to 0.1 μm and of the pores having diameters of greater than or equal to 0.2 4m is, unexpectedly and in a surprising proportion, given the prior knowledge, essential for achieving agglomerates with the maximum mechanical strength independently of the level of total porosity of the alumina.\",\n \"All the specimens according to the invention have a V37 Å of greater than 75 ml/100 g and a V0.1 μm of less than 31 ml/100 g. They also have a V0.2 μm of less than 20 ml/100 g. They also have a V1 μm of less than 7 ml/100 g and a V0.1 μm/V0.2 μm ratio of at least 1.5, in accordance with the preferred characteristics of the invention.\",\n \"However, it should be pointed out that the features that have been given as preferred are independent of one another and that it would remain in accordance with the invention to comply with some of them without complying with the others.\",\n \"It should be noted that the control specimens 9 to 11 all have a high V37 Å, in accordance with one of the obligatory characteristics of the aluminas of the invention.\",\n \"However, they have a V0.1 μm and a V0.2 μm that are higher than the maximum required by the invention, coupled with a V1 μm of greater than the preferred upper limit of the invention.\",\n \"This means that these specimens have a quantity of large-diameter pores that is relatively higher than that required by the invention.\",\n \"However, these specimens have only relatively mediocre mechanical properties as regards crush strength (Spp) and resistance to friction (shear).\",\n \"This major influence of the presence of large-diameter pores is also found when the results of specimens 1 to 8 according to the invention are examined.\",\n \"The best mechanical strength results are obtained for specimens 7 and 8, the V37 Å of which is not exceptionally high, and these specimens are distinguished from the others by particularly low V0.1 μm, V0.2 μm and V1 μm values.\",\n \"It has been found that this pore distribution gives these specimens a crush strength very substantially higher than that of the other specimens, although their shear strength is at least very good.\",\n \"The best results are obtained with specimen 7 that has the lowest V0.1 μm and V0.2 μm values of the series.\",\n \"It may also be noted that specimens (1, 2, 6) having a very high V37 Å, and therefore a very high overall porosity, but V0.1 μm, V0.2 μm, V1 μm and V0.1 μm/V0.2 μm values within the necessary or preferred limits of the invention, nevertheless have mechanical properties that are still substantially better than those of the control specimens.\",\n \"However, these have a lower overall porosity that would a priori lead to a better mechanical strength.\",\n \"Here again, controlling the presence of pores having diameters of greater than 0.1 μm, or even 0.2 μm and 1 μm, under the specified conditions appears to be particularly advantageous.\",\n \"Measuring the V0.1 μm/V0.2 μm ratio gives an indicator of what the distribution of the pore diameters in the 0.1-0.2 μm range should be.\",\n \"The invention is applicable for obtaining catalyst supports in the form of beads, especially those for the petroleum industry and the petrochemical industry, in which the combination of a high porosity and a high mechanical strength of the supports gives effective catalysts with a long lifetime.\",\n \"These catalysts may be used, for example, in hydrotreatment, particularly in hydrometallation, to reduce the metal content of a hydrocarbon cut.\",\n \"These alumina agglomerates can also be used as intrinsic catalysts or as adsorbents.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473938"}}},{"rowIdx":767,"cells":{"text":{"kind":"list like","value":[["Adjustment Method, Particularly a Laser Adjustment Method, and an Actuator Suitable for the Same","The invention relates to an adjustment method, especially for the adjustment of optical or fibre optical components, whereby a partial region of an actuator is locally heated in such a defined manner that compressive stresses are created therein as a result of the restricted thermal expansion of said heated partial region, caused by at least one other partial region of the actuator.","If the yielding point σ of the material of the partial region is exceeded, said compressive stresses lead to the plastic compression of the heated partial region.","Furthermore, said heated partial region is contracted during cooling and leads to a defined modification of the geometry of the actuator following the cooling process.","Tensile stresses are created as a result of a restriction of said contraction by means of at least one other partial region in the previously heated partial region, and compressive stresses are frozen in the at least one other region.","During the adjustment process, essential regions of the actuator, in which the tensile stresses or compressive stresses are frozen after cooling, are brought to a critical temperature in relation to the operating temperature of the actuator, at least until flow processes of the material are ended at the critical temperature."],["1-7.","(canceled) 8.A method for adjusting optical or fiber optic components, comprising: locally heating a selected subregion of an actuator such that compressive stresses arise in the selected subregion, which, upon reaching a yield point of the material of the selected subregion, results in plastic deformation of the selected subregion; and cooling the selected subregion, such that the selected subregion is shortened, resulting in a defined geometric change of the actuator, wherein tensile stresses arise in a first subregion of the actuator and, in a second region of the actuator, the compressive stresses are frozen, wherein: a region of the actuator in which the tensile stresses or compressive stresses exist after cooling is brought to a critical temperature with regard to a usage temperature range of the actuator, at least until flow processes of the material at the critical temperature are substantially concluded.","9.The method of claim 8, wherein the compressive stresses arise from thermal expansion of the selected subregion being substantially constrained by an adjacent subregion of the actuator.","10.The method of claim 8, wherein the critical temperature is selected to be essentially the temperature within the usage temperature range at which the minimum yield point of the material is attained.","11.The method of claim 8, wherein the material of the selected subregion of the actuator has a yield point that drops monotonically, at least within the usage temperature range.","12.The method of claim 11, wherein the critical temperature is selected to be essentially the temperature at an upper limit of the usage temperature range.","13.The method of claim 8, further comprising brining an entire subassembly that includes the actuator to the critical temperature.","14.The method of claim 8, wherein locally heating the selected subregion is performed using an Nd:YAG laser.","15.The method of claim 8, wherein locally heating the selected subregion is performed using a diode laser.","16.An actuator for adjusting an optical or a fiber optics component, comprising: means for locally heating a selected subregion of the actuator such that compressive stresses arise in the selected subregion, which, upon reaching a yield point of the material of the selected subregion, results in plastic deformation of the selected subregion; and means for cooling the selected subregion, such that the selected subregion is shortened, resulting in a defined geometric change of the actuator, wherein tensile stresses arise in a first subregion of the actuator and, in a second region of the actuator, the compressive stresses are frozen, wherein: a region of the actuator in which the tensile stresses or compressive stresses exist after cooling is brought to a critical temperature with regard to a usage temperature range of the actuator, at least until flow processes of the material at the critical temperature are substantially concluded.","17.The actuator of claim 16, wherein at least the selected subregion of the actuator is formed from a material having a yield point that assumes a minimum value within the entire usage temperature range essentially at room temperature.","18.The actuator of claim 16, wherein the means for locally heating the selected subregion comprises a laser.","19.The actuator of claim 18, wherein the laser comprises an Nd:YAG laser.","20.The actuator of claim 18, wherein the laser comprises a diode laser.","21.The actuator of claim 16, further comprising: a base; an adjustment region that moves in response to the means for locally heating and the means for cooling; and a pair of double bridges that connect the base and the adjustment region, wherein the selected subregion is included in on of the bridges of the pair of double bridges.","22.The actuator of claim 16, wherein the critical temperature is selected to be essentially the temperature within the usage temperature range at which the minimum yield point of the material is attained.","23.The actuator of claim 16, wherein the material of the selected subregion has a yield point that drops monotonically within the usage temperature range.","24.The actuator of claim 23, wherein the critical temperature is selected to be essentially the temperature at an upper limit of the usage temperature range."],["The invention concerns an adjustment method, particularly for adjusting optical or fiber optical components, with the features of the preamble of patent claim 1.Moreover, the invention concerns an actuator which is suitable for the same.","Laser adjustment methods and actuators suitable for the same have been developed in recent times to enable extremely precise adjustment, e.g., of micromechanical components.","The basic principle of laser adjustment consists of heating a predetermined region of an actuator using a high-energy, preferably pulsed laser beam in a short time, in which the thermal expansion of the relevant region is blocked by corresponding further regions of the actuator.","In this manner, compression strains build up in this heated region, which result in a plastic deformation of this region upon reaching the yield point.","When this region is cooled down after the high-energy laser beam is switched off, the thermal shrinkage of this region is again essentially prevented by the further regions of the actuator.","This leads to the build-up of tensile stresses in the previously heated region which lead to a defined deformation of the actuator, in which the adjustment of a component joined to the actuator is enabled in this process.","The basic principle of such an adjustment method is illustrated in FIG.","1.FIG.","1a shows the starting state of a bar 3 made of a suitable material which is restrained between two rigid demarcations 1.At the starting temperature, e.g., room temperature (20° C.), the bar 3 has a length l0.The center region 3a of the bar 3 is heated by a laser beam 5 in a short time.","This results in the first place in compressive stresses σ(−) in the bar 3 since the thermal longitudinal expansion of the bar 3 is blocked by the rigid demarcations 1.In this phase, a negative expansion ε is also customarily defined which corresponds to the compressive stresses formed in the bar 3.This phase of the adjustment method is illustrated in FIG.","1b.","If the compressive stresses σ(−) exceed the temperature-dependent yield point σF, a plastic deformation of the bar 3 arises in the region 3a.","Correspondingly, the compressive stresses in the bar 3 are reduced.","This situation is illustrated in FIG.","1c.","After the laser beam 5 is switched off, the region 3a of the bar 3 begins to cool off, causing a thermal shrinkage of the bar 3.In this process, tensile stresses σ(+) arise in the bar 3 which in actual practice frequently lie in the vicinity of the temperature-dependent yield point σF(T).","This situation is represented symbolically in FIG.","1d because seen in the longitudinal direction of the bar 3 whose ends are joined by means of spring elements 7 to the rigid demarcations 1 .",".",".","[Note: incomplete sentence in the German original].","The spring forces corresponding to the tensile stresses σ(+) cause a defined deformation of the actuator in a practically realized actuator.","In this connection, FIG.","1d also shows that the tensile stresses σ(+) arise through the shortening of the bar 3 due to the plastic deformation in the region 3a caused during the heating, in which the length of the bar l1 after the adjustment procedure at the starting temperature is smaller than the original length l0 at the starting temperature.","A problem in the previously known adjustment methods consists in that, as was previously mentioned, the tensile stresses frozen in the bar 3 lie relatively close to the yield point σF.","The same can apply also to the compressive stresses which occur in those regions which block the thermal expansion or rather the thermal shrinkage of the bar.","Since micromechanical or optical components or rather subassemblies in actual practice are always specified for a certain temperature range, e.g., a range from −40° C. to +80° C., and must fulfill predetermined requirements for accuracy and long-term stability within the specified range, there results in previously known adjustment methods a maladjustment of the actuator if the adjusted components or rather the subassembly is brought to a temperature in the upper region of the specified range and the original adjustment was carried out at a significantly lower temperature, such as room temperature.","This effect is caused by the temperature dependency of the yield point σF, most materials which are suitable for the manufacture of actuators for laser adjustment methods having a yield point which decreases with increasing temperature.","If a temperature is reached at which the yield point σF falls below the value of the frozen-in tensile stresses, this results in a flowing of the material and in a reduction of the tensile stresses to the value of the yield point σF at the relevant temperature.","Naturally, this is associated with a corresponding maladjustment of the actuator which is not acceptable at least for components requiring extremely precise adjustment which must be specified over a wide temperature range.","The underlying object of the invention is therefore to create an adjustment method, particularly for adjusting optical or fiber optical components, with which improved long-term stability of an adjusted actuator or rather a subassembly having such an actuator can be ensured within a predetermined temperature range, it being possible to carry out the method quickly and at a low cost.","Moreover, the underlying object of the invention is to create a special actuator for simply carrying out the method.","This objective is solved by the invention with the features of patent claim 1 or 6.The invention is based on the insight that the adjustment of an actuator can be carried out in an advantageous manner at a critical temperature Tk with regard to the predetermined usage temperature range of the actuator.","In this connection, a critical temperature Tk is understood to be a temperature at which the yield point has a value such that when passing through the entire usage temperature range, only minor maladjustments of the adjusted actuator can occur which lie within specified tolerances.","With conventional actuator materials, which have within a specification range a yield point that decreases at higher temperatures, one will choose the critical temperature in the upper region of the specification range or even a temperature lying above its upper limit To.","According to an embodiment of the invention, the critical temperature Tk can essentially be chosen to be that temperature within the usage temperature range at which the minimum yield point σF of the material is attained within the usage temperature range.","In this case, it is ensured that within the usage temperature range, no flowing of the material occurs in those regions of the actuator in which internal stresses occur as a result of the adjustment procedure.","The possibility explained above of selecting an even higher temperature is limited in actual practice by the fact that often such components are also contained in subassemblies which may be subjected as a maximum temperature to the upper temperature To of the usage temperature range.","According to an embodiment of the method, an entire subassembly including the actuator can be brought to the critical temperature Tk during the adjustment procedure.","If only subregions of the actuator are brought to the critical temperature Tk during the adjustment procedure, at least those regions must be included which are subject to thermal shrinkage but also those which prevent such shrinkage.","A special actuator with which the method according to the invention can be implemented in a particularly simple manner is formed such that at least the essential regions of the actuator consist of a material whose yield point σF assumes a minimum value within the entire usage temperature range essentially at room temperature.","Using an actuator of this sort, an adjustment procedure can be carried out as usual at room temperature.","According to an embodiment of the invention, the actuator or rather the essential regions of the actuator can consist of a material whose yield point does not drop or rather remains essentially constant over the entire usage temperature range.","For example, TiV13Cr11A13 can be used as a material for the actuator or rather the essential regions of the actuator.","The 0.2% permanent elongation limit, resistance to extension and yield point of this material does not decrease measurably up to a temperature of 100° C. Further embodiments of the invention follow from the dependent claims.","The invention is described hereafter in greater detail based on the figures shown in the drawing.","The figures are as follows: FIG.","1: Schematic representations of the laser beam adjustment method; FIG.","2: An embodiment of a practical actuator having two double bridges; and FIG.","3: A chart showing the temperature-dependent yield point σF(T) in a conventional actuator material within a usage temperature range [Tu;To].","The laser beam adjustment method explained in basic terms in the introduction is illustrated again briefly in greater detail based on a practical actuator 10 shown in FIG.","2.The actuator 10 consists of a base region 12 which is joined by means of two double bridges I, II to an adjustment region 14.For adjustment purposes, in each case one of the two bridges of the double bridges I, II is irradiated preferably in the center (in relation to the longitudinal axis of a bridge) using a high-power laser beam.","For example, if the bridge shown to the left in FIG.","2 of the double bridge I is heated in its center region using the laser beam, then the affected bridge attempts to expand, the thermal expansion being essentially blocked by the right bridge of the double bridge I.","In this process, the left double bridge is deformed in its center region.","During the cooling, the left bridge of the double bridge I attempts to draw together, through which corresponding tensile stresses arise in it in this process, since this movement is also blocked largely by the right bridge.","If in a next step the center region of the bridge shown to the right in FIG.","2 of the double bridge I is also irradiated using the laser, then this region will be deformed due to the tensile stresses present in the left bridge and the compressive stress present in the right bridge (in addition to the compressive stresses which are caused by a thermal expansion of the right bridge).","During the cooling, the right bridge of the double bridge I attempts to draw together further, such that this movement is blocked in turn by the left bridge.","Accordingly, internal stresses remain, which are formed in the right bridge as tensile stresses and in the left bridge as compressive stresses.","As a result of this adjustment procedure, the adjustment region 14 is tilted slightly with respect to the starting position shown in FIG.","2 using a continuous line as a result of the shortening of the two bridges of the double bridge I.","This situation is illustrated in FIG.","2 using a dashed line.","In this manner, an adjustment of the angle of a component (not shown) arranged on the adjustment region 14 can occur.","If the right double bridge II is also adjusted correspondingly, then highly precise adjustment of a component arranged on the adjustment region 14 can occur in the longitudinal direction of the bridges.","If the adjustment procedures are carried out at an critical temperature Tk, in which it is necessary for at least the double bridge to be adjusted or rather for both double bridges to be adjusted to be at this temperature, then upon suitable choice of the critical temperature Tk only such internal stresses are frozen in as a consequence of the adjustment procedure(s) which, even in case of longer-term storage of the actuator 10 at an arbitrary temperature within a predetermined usage temperature range [Tu;To], lead to such maladjustments that lie within acceptable tolerances.","If, for example, the actuator material exhibits the curve of its temperature-dependent yield point σF(T) shown in FIG.","3 as a continuous line, then one will choose, as shown in FIG.","3, the critical temperature Tk in the upper region of the usage temperature range [Tu;To].","The critical temperature Tk must be chosen for the monotonically decreasing curve of the yield point σF(T) shown in FIG.","3 to be so high that even in case of longer-term storage of the actuator at the upper limit Tu of the usage temperature range only such a maladjustment will occur as a consequence of a reduction in internal stresses in the essential regions of the actuator that lies within specified permissible tolerances.","If one assumes that in the essential regions of the actuator internal stresses are frozen in which lie extremely close to the yield point at the critical temperature or rather the adjustment temperature chosen in each case, then a maladjustment in case of longer-term storage of the actuator at the temperature To can be avoided in full only if the critical temperature Tk is chosen to be equal to the upper range limit To.","If a material is chosen for the actuator or its essential regions whose yield point σF(T) over the entire usage temperature range [Tu;To] is essentially constant, then the critical temperature Tk can be chosen arbitrarily.","Here, one will preferably choose the temperature Tk in the room temperature range to achieve the lowest cost.","Finally, it should be mentioned that the heating of the essential regions of the actuator, the overall actuator or an overall subassembly including the actuator can take place, for example, through irradiation with an infrared radiation source by using a customary, heated temperature chamber or a heated holder, or even using a laser beam which, suitably expanded, is either aimed at the essential regions or which “scans” the essential regions at a suitable speed."]],"string":"[\n [\n \"Adjustment Method, Particularly a Laser Adjustment Method, and an Actuator Suitable for the Same\",\n \"The invention relates to an adjustment method, especially for the adjustment of optical or fibre optical components, whereby a partial region of an actuator is locally heated in such a defined manner that compressive stresses are created therein as a result of the restricted thermal expansion of said heated partial region, caused by at least one other partial region of the actuator.\",\n \"If the yielding point σ of the material of the partial region is exceeded, said compressive stresses lead to the plastic compression of the heated partial region.\",\n \"Furthermore, said heated partial region is contracted during cooling and leads to a defined modification of the geometry of the actuator following the cooling process.\",\n \"Tensile stresses are created as a result of a restriction of said contraction by means of at least one other partial region in the previously heated partial region, and compressive stresses are frozen in the at least one other region.\",\n \"During the adjustment process, essential regions of the actuator, in which the tensile stresses or compressive stresses are frozen after cooling, are brought to a critical temperature in relation to the operating temperature of the actuator, at least until flow processes of the material are ended at the critical temperature.\"\n ],\n [\n \"1-7.\",\n \"(canceled) 8.A method for adjusting optical or fiber optic components, comprising: locally heating a selected subregion of an actuator such that compressive stresses arise in the selected subregion, which, upon reaching a yield point of the material of the selected subregion, results in plastic deformation of the selected subregion; and cooling the selected subregion, such that the selected subregion is shortened, resulting in a defined geometric change of the actuator, wherein tensile stresses arise in a first subregion of the actuator and, in a second region of the actuator, the compressive stresses are frozen, wherein: a region of the actuator in which the tensile stresses or compressive stresses exist after cooling is brought to a critical temperature with regard to a usage temperature range of the actuator, at least until flow processes of the material at the critical temperature are substantially concluded.\",\n \"9.The method of claim 8, wherein the compressive stresses arise from thermal expansion of the selected subregion being substantially constrained by an adjacent subregion of the actuator.\",\n \"10.The method of claim 8, wherein the critical temperature is selected to be essentially the temperature within the usage temperature range at which the minimum yield point of the material is attained.\",\n \"11.The method of claim 8, wherein the material of the selected subregion of the actuator has a yield point that drops monotonically, at least within the usage temperature range.\",\n \"12.The method of claim 11, wherein the critical temperature is selected to be essentially the temperature at an upper limit of the usage temperature range.\",\n \"13.The method of claim 8, further comprising brining an entire subassembly that includes the actuator to the critical temperature.\",\n \"14.The method of claim 8, wherein locally heating the selected subregion is performed using an Nd:YAG laser.\",\n \"15.The method of claim 8, wherein locally heating the selected subregion is performed using a diode laser.\",\n \"16.An actuator for adjusting an optical or a fiber optics component, comprising: means for locally heating a selected subregion of the actuator such that compressive stresses arise in the selected subregion, which, upon reaching a yield point of the material of the selected subregion, results in plastic deformation of the selected subregion; and means for cooling the selected subregion, such that the selected subregion is shortened, resulting in a defined geometric change of the actuator, wherein tensile stresses arise in a first subregion of the actuator and, in a second region of the actuator, the compressive stresses are frozen, wherein: a region of the actuator in which the tensile stresses or compressive stresses exist after cooling is brought to a critical temperature with regard to a usage temperature range of the actuator, at least until flow processes of the material at the critical temperature are substantially concluded.\",\n \"17.The actuator of claim 16, wherein at least the selected subregion of the actuator is formed from a material having a yield point that assumes a minimum value within the entire usage temperature range essentially at room temperature.\",\n \"18.The actuator of claim 16, wherein the means for locally heating the selected subregion comprises a laser.\",\n \"19.The actuator of claim 18, wherein the laser comprises an Nd:YAG laser.\",\n \"20.The actuator of claim 18, wherein the laser comprises a diode laser.\",\n \"21.The actuator of claim 16, further comprising: a base; an adjustment region that moves in response to the means for locally heating and the means for cooling; and a pair of double bridges that connect the base and the adjustment region, wherein the selected subregion is included in on of the bridges of the pair of double bridges.\",\n \"22.The actuator of claim 16, wherein the critical temperature is selected to be essentially the temperature within the usage temperature range at which the minimum yield point of the material is attained.\",\n \"23.The actuator of claim 16, wherein the material of the selected subregion has a yield point that drops monotonically within the usage temperature range.\",\n \"24.The actuator of claim 23, wherein the critical temperature is selected to be essentially the temperature at an upper limit of the usage temperature range.\"\n ],\n [\n \"The invention concerns an adjustment method, particularly for adjusting optical or fiber optical components, with the features of the preamble of patent claim 1.Moreover, the invention concerns an actuator which is suitable for the same.\",\n \"Laser adjustment methods and actuators suitable for the same have been developed in recent times to enable extremely precise adjustment, e.g., of micromechanical components.\",\n \"The basic principle of laser adjustment consists of heating a predetermined region of an actuator using a high-energy, preferably pulsed laser beam in a short time, in which the thermal expansion of the relevant region is blocked by corresponding further regions of the actuator.\",\n \"In this manner, compression strains build up in this heated region, which result in a plastic deformation of this region upon reaching the yield point.\",\n \"When this region is cooled down after the high-energy laser beam is switched off, the thermal shrinkage of this region is again essentially prevented by the further regions of the actuator.\",\n \"This leads to the build-up of tensile stresses in the previously heated region which lead to a defined deformation of the actuator, in which the adjustment of a component joined to the actuator is enabled in this process.\",\n \"The basic principle of such an adjustment method is illustrated in FIG.\",\n \"1.FIG.\",\n \"1a shows the starting state of a bar 3 made of a suitable material which is restrained between two rigid demarcations 1.At the starting temperature, e.g., room temperature (20° C.), the bar 3 has a length l0.The center region 3a of the bar 3 is heated by a laser beam 5 in a short time.\",\n \"This results in the first place in compressive stresses σ(−) in the bar 3 since the thermal longitudinal expansion of the bar 3 is blocked by the rigid demarcations 1.In this phase, a negative expansion ε is also customarily defined which corresponds to the compressive stresses formed in the bar 3.This phase of the adjustment method is illustrated in FIG.\",\n \"1b.\",\n \"If the compressive stresses σ(−) exceed the temperature-dependent yield point σF, a plastic deformation of the bar 3 arises in the region 3a.\",\n \"Correspondingly, the compressive stresses in the bar 3 are reduced.\",\n \"This situation is illustrated in FIG.\",\n \"1c.\",\n \"After the laser beam 5 is switched off, the region 3a of the bar 3 begins to cool off, causing a thermal shrinkage of the bar 3.In this process, tensile stresses σ(+) arise in the bar 3 which in actual practice frequently lie in the vicinity of the temperature-dependent yield point σF(T).\",\n \"This situation is represented symbolically in FIG.\",\n \"1d because seen in the longitudinal direction of the bar 3 whose ends are joined by means of spring elements 7 to the rigid demarcations 1 .\",\n \".\",\n \".\",\n \"[Note: incomplete sentence in the German original].\",\n \"The spring forces corresponding to the tensile stresses σ(+) cause a defined deformation of the actuator in a practically realized actuator.\",\n \"In this connection, FIG.\",\n \"1d also shows that the tensile stresses σ(+) arise through the shortening of the bar 3 due to the plastic deformation in the region 3a caused during the heating, in which the length of the bar l1 after the adjustment procedure at the starting temperature is smaller than the original length l0 at the starting temperature.\",\n \"A problem in the previously known adjustment methods consists in that, as was previously mentioned, the tensile stresses frozen in the bar 3 lie relatively close to the yield point σF.\",\n \"The same can apply also to the compressive stresses which occur in those regions which block the thermal expansion or rather the thermal shrinkage of the bar.\",\n \"Since micromechanical or optical components or rather subassemblies in actual practice are always specified for a certain temperature range, e.g., a range from −40° C. to +80° C., and must fulfill predetermined requirements for accuracy and long-term stability within the specified range, there results in previously known adjustment methods a maladjustment of the actuator if the adjusted components or rather the subassembly is brought to a temperature in the upper region of the specified range and the original adjustment was carried out at a significantly lower temperature, such as room temperature.\",\n \"This effect is caused by the temperature dependency of the yield point σF, most materials which are suitable for the manufacture of actuators for laser adjustment methods having a yield point which decreases with increasing temperature.\",\n \"If a temperature is reached at which the yield point σF falls below the value of the frozen-in tensile stresses, this results in a flowing of the material and in a reduction of the tensile stresses to the value of the yield point σF at the relevant temperature.\",\n \"Naturally, this is associated with a corresponding maladjustment of the actuator which is not acceptable at least for components requiring extremely precise adjustment which must be specified over a wide temperature range.\",\n \"The underlying object of the invention is therefore to create an adjustment method, particularly for adjusting optical or fiber optical components, with which improved long-term stability of an adjusted actuator or rather a subassembly having such an actuator can be ensured within a predetermined temperature range, it being possible to carry out the method quickly and at a low cost.\",\n \"Moreover, the underlying object of the invention is to create a special actuator for simply carrying out the method.\",\n \"This objective is solved by the invention with the features of patent claim 1 or 6.The invention is based on the insight that the adjustment of an actuator can be carried out in an advantageous manner at a critical temperature Tk with regard to the predetermined usage temperature range of the actuator.\",\n \"In this connection, a critical temperature Tk is understood to be a temperature at which the yield point has a value such that when passing through the entire usage temperature range, only minor maladjustments of the adjusted actuator can occur which lie within specified tolerances.\",\n \"With conventional actuator materials, which have within a specification range a yield point that decreases at higher temperatures, one will choose the critical temperature in the upper region of the specification range or even a temperature lying above its upper limit To.\",\n \"According to an embodiment of the invention, the critical temperature Tk can essentially be chosen to be that temperature within the usage temperature range at which the minimum yield point σF of the material is attained within the usage temperature range.\",\n \"In this case, it is ensured that within the usage temperature range, no flowing of the material occurs in those regions of the actuator in which internal stresses occur as a result of the adjustment procedure.\",\n \"The possibility explained above of selecting an even higher temperature is limited in actual practice by the fact that often such components are also contained in subassemblies which may be subjected as a maximum temperature to the upper temperature To of the usage temperature range.\",\n \"According to an embodiment of the method, an entire subassembly including the actuator can be brought to the critical temperature Tk during the adjustment procedure.\",\n \"If only subregions of the actuator are brought to the critical temperature Tk during the adjustment procedure, at least those regions must be included which are subject to thermal shrinkage but also those which prevent such shrinkage.\",\n \"A special actuator with which the method according to the invention can be implemented in a particularly simple manner is formed such that at least the essential regions of the actuator consist of a material whose yield point σF assumes a minimum value within the entire usage temperature range essentially at room temperature.\",\n \"Using an actuator of this sort, an adjustment procedure can be carried out as usual at room temperature.\",\n \"According to an embodiment of the invention, the actuator or rather the essential regions of the actuator can consist of a material whose yield point does not drop or rather remains essentially constant over the entire usage temperature range.\",\n \"For example, TiV13Cr11A13 can be used as a material for the actuator or rather the essential regions of the actuator.\",\n \"The 0.2% permanent elongation limit, resistance to extension and yield point of this material does not decrease measurably up to a temperature of 100° C. Further embodiments of the invention follow from the dependent claims.\",\n \"The invention is described hereafter in greater detail based on the figures shown in the drawing.\",\n \"The figures are as follows: FIG.\",\n \"1: Schematic representations of the laser beam adjustment method; FIG.\",\n \"2: An embodiment of a practical actuator having two double bridges; and FIG.\",\n \"3: A chart showing the temperature-dependent yield point σF(T) in a conventional actuator material within a usage temperature range [Tu;To].\",\n \"The laser beam adjustment method explained in basic terms in the introduction is illustrated again briefly in greater detail based on a practical actuator 10 shown in FIG.\",\n \"2.The actuator 10 consists of a base region 12 which is joined by means of two double bridges I, II to an adjustment region 14.For adjustment purposes, in each case one of the two bridges of the double bridges I, II is irradiated preferably in the center (in relation to the longitudinal axis of a bridge) using a high-power laser beam.\",\n \"For example, if the bridge shown to the left in FIG.\",\n \"2 of the double bridge I is heated in its center region using the laser beam, then the affected bridge attempts to expand, the thermal expansion being essentially blocked by the right bridge of the double bridge I.\",\n \"In this process, the left double bridge is deformed in its center region.\",\n \"During the cooling, the left bridge of the double bridge I attempts to draw together, through which corresponding tensile stresses arise in it in this process, since this movement is also blocked largely by the right bridge.\",\n \"If in a next step the center region of the bridge shown to the right in FIG.\",\n \"2 of the double bridge I is also irradiated using the laser, then this region will be deformed due to the tensile stresses present in the left bridge and the compressive stress present in the right bridge (in addition to the compressive stresses which are caused by a thermal expansion of the right bridge).\",\n \"During the cooling, the right bridge of the double bridge I attempts to draw together further, such that this movement is blocked in turn by the left bridge.\",\n \"Accordingly, internal stresses remain, which are formed in the right bridge as tensile stresses and in the left bridge as compressive stresses.\",\n \"As a result of this adjustment procedure, the adjustment region 14 is tilted slightly with respect to the starting position shown in FIG.\",\n \"2 using a continuous line as a result of the shortening of the two bridges of the double bridge I.\",\n \"This situation is illustrated in FIG.\",\n \"2 using a dashed line.\",\n \"In this manner, an adjustment of the angle of a component (not shown) arranged on the adjustment region 14 can occur.\",\n \"If the right double bridge II is also adjusted correspondingly, then highly precise adjustment of a component arranged on the adjustment region 14 can occur in the longitudinal direction of the bridges.\",\n \"If the adjustment procedures are carried out at an critical temperature Tk, in which it is necessary for at least the double bridge to be adjusted or rather for both double bridges to be adjusted to be at this temperature, then upon suitable choice of the critical temperature Tk only such internal stresses are frozen in as a consequence of the adjustment procedure(s) which, even in case of longer-term storage of the actuator 10 at an arbitrary temperature within a predetermined usage temperature range [Tu;To], lead to such maladjustments that lie within acceptable tolerances.\",\n \"If, for example, the actuator material exhibits the curve of its temperature-dependent yield point σF(T) shown in FIG.\",\n \"3 as a continuous line, then one will choose, as shown in FIG.\",\n \"3, the critical temperature Tk in the upper region of the usage temperature range [Tu;To].\",\n \"The critical temperature Tk must be chosen for the monotonically decreasing curve of the yield point σF(T) shown in FIG.\",\n \"3 to be so high that even in case of longer-term storage of the actuator at the upper limit Tu of the usage temperature range only such a maladjustment will occur as a consequence of a reduction in internal stresses in the essential regions of the actuator that lies within specified permissible tolerances.\",\n \"If one assumes that in the essential regions of the actuator internal stresses are frozen in which lie extremely close to the yield point at the critical temperature or rather the adjustment temperature chosen in each case, then a maladjustment in case of longer-term storage of the actuator at the temperature To can be avoided in full only if the critical temperature Tk is chosen to be equal to the upper range limit To.\",\n \"If a material is chosen for the actuator or its essential regions whose yield point σF(T) over the entire usage temperature range [Tu;To] is essentially constant, then the critical temperature Tk can be chosen arbitrarily.\",\n \"Here, one will preferably choose the temperature Tk in the room temperature range to achieve the lowest cost.\",\n \"Finally, it should be mentioned that the heating of the essential regions of the actuator, the overall actuator or an overall subassembly including the actuator can take place, for example, through irradiation with an infrared radiation source by using a customary, heated temperature chamber or a heated holder, or even using a laser beam which, suitably expanded, is either aimed at the essential regions or which “scans” the essential regions at a suitable speed.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473940"}}},{"rowIdx":768,"cells":{"text":{"kind":"list like","value":[["Adjustment Method, Particularly a Laser Adjustment Method, and an Actuator Suitable For the Same","The invention relates to an adjustment method, especially for adjusting optical or fibre optical components.","Said method involves locally heating, in a defined manner, at least one first partial region of an adjustment region (106, 114) of an actuator (100, 200, 400), in such a way that compressive strains appear in at least one second partial region of the adjustment region of the actuator (100, 200, 400) as a result of the fact that said second partial region prevents the thermal expansion of the heated partial area.","When the yielding point σF of the material of the partial region is achieved, said compressive strains lead to a plastic deformation of the heated partial region.","The heated first partial region shrinks during the cooling process, triggering a defined geometry modification of the actuator (100, 200, 400) following the cooling process.","Due to the fact that the at least one second partial region prevents the shrinking, tensile stresses appear in the previously heated first partial region and the compressive strains are frozen in the at least one second region.","According to the invention, the regions of the actuator (100, 200, 300, 400), in which the tensile stresses or compressive strains are frozen following the cooling process, are brought to a critical temperature (Tk) in relation to the operational temperature range of the actuator, after the cooling process, at least until the flow processes of the material at said critical temperature are largely completed.","A second adjustment process is then carried out.","The invention also relates to an actuator which is especially suitable for carrying out the inventive method, and an optical component comprising one such actuator."],["1-22.","(canceled) 23.A method for adjusting optical or fiber optic components, comprising the acts of: performing a first adjustment procedure, including: locally heating a selected subregion of an actuator such that compressive stresses arise in the selected subregion, which, upon reaching a yield point of the material of the selected subregion, results in plastic deformation of the selected subregion; cooling the selected subregion, such that the selected subregion is shortened, resulting in a defined geometric change of the actuator, wherein tensile stresses arise in a first subregion of the actuator and, in a second region of the actuator, the compressive stresses are frozen; and reheating the selected subregion to a specified temperature such that stresses in a portion thereof are relieved; and performing a second adjustment procedure.","24.The method of claim 23, wherein second adjustment procedure comprises locally heating and cooling another subregion of the actuator, wherein the other subregion is positioned, such that, in comparison to the selected subregion, a smaller adjustment effect is caused if the same amount of energy is supplied.","25.The method of claim 23, wherein, after the first adjustment procedure, the selected subregion is brought to a critical temperature with regard to the usage temperature range of the actuator, at least until flow processes of the material are substantially concluded at the critical temperature.","26.The method of claim 25, wherein the critical temperature is selected to be essentially the temperature within the usage temperature range at which a minimum yield point of the material is attained within the usage temperature range.","27.The method of claim 25, wherein the material of the selected subregion has a yield point that drops monotonically with increasing temperatures within the usage temperature range 28.The method of claim 27, wherein the critical temperature is selected to be essentially the temperature corresponding to an upper limit of the usage temperature range.","29.The method of claim 23, wherein locally heating the selected subregion is performed using an Nd:YAG laser.","30.The method of claim 23, wherein locally heating the selected subregion is performed using a diode laser.","31.An actuator for adjusting an optical or a fiber optics component, comprising: a support region for accommodating the component to be adjusted; a first adjustment region having a double bridge; and a second adjustment region having a simple bridge, wherein the first adjustment region produces a greater adjustment effect when an adjustment procedure is performed using the first adjustment region than an adjustment effect produced when an adjustment procedure is performed using the second adjustment region.","32.The actuator of claim 31, further comprising means for heating a first bridge of the double bridge, wherein thermal expansion of the first bridge of the double bridge is constrained by the second bridge of the double bridge.","33.The actuator of claim 32, further comprising means for heating a first subregion of the simple bridge, wherein a second subregion of the simple bridge constrains thermal expansion of the first subregion of the simple bridge.","34.The actuator of claim 31, further comprising a substantially cylindrical body having an end at which the support region is positioned.","35.The actuator of claim 34, wherein the first adjustment region and the second adjustment region are positioned in the cylindrical body.","36.The actuator of claim 35, wherein the first adjustment region is positioned to be closer to the support region than is the second adjustment region.","37.The actuator of claim 35, wherein first adjustment region defines a plane that is substantially perpendicular to an axis of the cylindrical body and substantially coincides with a plane that forms the basis of an adjustment procedure that can be performed using the first adjustment region.","38.The actuator of claim 35 wherein: the first adjustment region includes three double bridges positioned about a circumference of the cylindrical body; and the second adjustment region includes three simple bridges positioned about the circumference of the cylindrical body.","39.A method of manufacturing an actuator for use with an optical or a fiber optics component, comprising: obtaining a substantially planar element having: a central region; and a plurality of at least three arms that are connected to and extend radially away from the central region; and bending the arms to form substantially right angles with respect to the central region, the bent arms defining a first adjustment region that defines a plane substantially parallel to the central region and a second adjustment region substantially parallel to the central region.","40.The method of claim 39, wherein the plurality of at least three arms comprises exactly three arms at an angular spacing of 120°.","41.The method of claim 40, wherein each of the plurality of at least three arms includes a double bridge that defines the first adjustment region and a simple bridge that defines the second adjustment region.","42.The method of claim 39, wherein the plurality of at least three arms comprises exactly six arms at an angular spacing of 60°.","43.The method of claim 42, wherein bending the arms comprises: bending the six arms in the same direction; and connecting adjacent the longitudinal sides of the six arms to form a hollow-cylindrical wall of the actuator.","44.The method of claim 43, wherein alternating arms of the six arms include a double bridge that defines the first adjustment region and a simple bridge that defines the second adjustment region."],["The invention concerns an adjustment method, particularly for adjusting optical or fiber optical components, with the features of the preamble of patent claim 1.Moreover, the invention concerns an actuator which is suitable for the same and an optical component.","Laser adjustment methods and actuators suitable for the same have been developed in recent times to enable extremely precise adjustment, e.g., of micromechanical components.","The basic principle of laser adjustment consists of heating a predetermined region of an actuator using a high-energy, preferably pulsed laser beam in a short time, in which the thermal expansion of the relevant region is blocked by corresponding further regions of the actuator.","In this manner, compression strains build up in this heated region, which result in a plastic deformation of this region upon reaching the yield point.","When this region is cooled down after the high-energy laser beam is switched off, the thermal shrinkage of this region is again essentially prevented by the further regions of the actuator.","This leads to the build-up of tensile stresses in the previously heated region which lead to a defined deformation of the actuator, in which the adjustment of a component joined to the actuator is enabled in this process.","The basic principle of such an adjustment method is illustrated in FIG.","1.FIG.","1a shows the starting state of a bar 3 made of a suitable material which is restrained between two rigid demarcations 1.At the starting temperature, e.g., room temperature (20° C.), the bar 3 has a length l0.The center region 3a of the bar 3 is heated by a laser beam 5 in a short time.","This results in the first place in compressive strains σ(−) in the bar 3 since the thermal longitudinal expansion of the bar 3 is blocked by the rigid demarcations 1.In this phase, a negative expansion ε is also customarily defined which corresponds to the compressive strains formed in the bar 3.This phase of the adjustment method is illustrated in FIG.","1b.","If the compressive strains σ(−) exceed the temperature-dependent yield point σF, a plastic deformation of the bar 3 arises in the region 3a.","Correspondingly, the compressive strains in the bar 3 are reduced.","This situation is illustrated in FIG.","1c.","After the laser beam 5 is switched off, the region 3a of the bar 3 begins to cool off, causing a thermal shrinkage of the bar 3.In this process, tensile stresses σ(+) arise in the bar 3 which in actual practice frequently lie in the vicinity of the temperature-dependent yield point σF(T).","This situation is represented symbolically in FIG.","1d because seen in the longitudinal direction of the bar 3 whose ends are joined by means of spring elements 7 to the rigid demarcations 1 .",".",".",".","[Note: incomplete sentence in the German original].","The spring forces corresponding to the tensile stresses σ(+) cause a defined deformation of the actuator in a practically realized actuator.","In this connection, FIG.","1d also shows that the tensile stresses σ(+) arise through the shortening of the bar 3 due to the plastic deformation in the region 3a caused during the heating, in which the length of the bar l1 after the adjustment procedure at the starting temperature is smaller than the original length l0 at the starting temperature.","A problem in the previously known adjustment methods consists in that, as was previously mentioned, the tensile stresses frozen in the bar 3 lie relatively close to the yield point σF.","The same can apply also to the compressive strains which occur in those regions which block the thermal expansion or rather the thermal shrinkage of the bar.","Since micromechanical or optical components or rather subassemblies in actual practice are always specified for a certain temperature range, e.g., a range from −40° C. to +80° C., and must fulfill predetermined requirements for accuracy and long-term stability within the specified range, there results in previously known adjustment methods a maladjustment of the actuator if the adjusted components or rather the subassembly is brought to a temperature in the upper region of the specified range and the original adjustment was carried out at a significantly lower temperature, such as room temperature.","This effect is caused by the temperature dependency of the yield point σF, most materials which are suitable for the manufacture of actuators for laser adjustment methods having a yield point which decreases with increasing temperature.","If a temperature is reached at which the yield point σF falls below the value of the frozen-in tensile stresses, this results in a flowing of the material and in a reduction of the tensile stresses to the value of the yield point σF at the relevant temperature.","Naturally, this is associated with a corresponding maladjustment of the actuator which is not acceptable at least for components requiring extremely precise adjustment which must be specified over a wide temperature range.","The underlying object of the invention is therefore to create an adjustment method, particularly for adjusting optical or fiber optical components, with which improved long-term stability of an adjusted actuator or rather a subassembly having such an actuator can be ensured within a predetermined temperature range, it being possible to carry out the method quickly and at a low cost.","Moreover, the underlying object of the invention is to create a special actuator for simply carrying out the method and an optical component having an actuator of this sort.","This objective is solved by the invention with the features of patent claim 1, 6 or 17.The invention is based on the insight that the adjustment of an actuator can be carried out in an advantageous manner in the first place in a first adjustment procedure at an arbitrary temperature, e.g., room temperature, that subsequently at least the essential regions of the actuator (in which stresses are frozen in as a consequence of the first adjustment procedure) can have their tension relieved by heating to a specified temperature, and that subsequently a readjustment of the actuator can be carried out again at an arbitrary temperature, preferably room temperature.","In an embodiment of the method, as regions essential for relieving the tension, all regions in which, after the first adjustment procedure compressive strains or tensile stresses are frozen in, are brought to a critical temperature (Tk) with regard to the usage temperature range of the actuator, at least until flow processes of the material are largely concluded at the critical temperature.","In this connection, a critical temperature (Tk) is understood to be a temperature at which the yield point has a value such that when passing through the entire usage temperature range, only minor maladjustments of the adjusted actuator can occur which lie within specified tolerances.","With conventional actuator materials which have within a specification range a yield point which decreases at higher temperatures, one will choose the critical temperature in the upper region of the specification range or even a temperature lying above its upper limit To.","According to the preferred embodiment, the second adjustment procedure is carried out by heating and cooling a first subregion of a further adjustment region of the actuator, the further adjustment region being constituted such that in comparison to the first adjustment region, a smaller adjustment effect would be caused if the same amount of energy is supplied.","Naturally, the energy of the second adjustment procedure or rather the curve vs. time of the power for heating the relevant region can differ in the second adjustment procedure and can be adapted particularly to the desired adjustment effect or rather adjustment range.","According to an embodiment of the invention, the critical temperature Tk can essentially be chosen to be that temperature within the usage temperature range at which the minimum yield point σF of the material is attained within the usage temperature range.","In this case, it is ensured that within the usage temperature range, no flowing of the material occurs in those regions of the actuator in which internal stresses occur as a result of the adjustment procedure.","The possibility explained above of choosing an even higher temperature assumes in actual practice that a corresponding heating of the essential regions is carried out only locally.","This applies at least if the actuator is part of a subassembly in which components are also contained which may be subjected as a maximum temperature to the upper temperature T0 of the planned usage temperature range.","The essential regions include those regions which are subject during an adjustment procedure to thermal shrinkage and also those regions which prevent such shrinkage.","In another embodiment of the method, essentially only those regions can be heated to a specified stress-relieving temperature as essential regions in which compressive strains are frozen in after the first adjustment procedure, in which the stress-relieving temperature is chosen such that a thermal expansion of the relevant regions [is] produced by the heating [and] the tensile stresses in those regions in which tensile stresses are frozen in following the first adjustment procedure, are increased above the yield point so that flowing of the material occurs in these regions.","In other words, through the heating of the regions with compressive strains and the resultant thermal expansion of these regions, a stretching of the regions with tensile stresses is carried out.","The stress-relieving temperature is chosen preferably such that after the tension is relieved during a reheating of all regions in which stresses are frozen in to a critical temperature (cf., in this connection the explanations above), no flow processes and thus no maladjustment of the actuator occurs.","The heating for the actual adjustment procedure and the heating for the heating after the adjustment procedure can take place, for example, using a laser, preferably an Nd:YAG laser or diode laser.","A special actuator with which the method according to the invention can be implemented in a particularly easy manner has a support region for accommodating an element to be adjusted and has at least a first and a second adjustment region, in which the first adjustment region is formed so that when carrying out an adjustment procedure using the first adjustment region, a greater adjustment effect can be obtained than when carrying out a further adjustment procedure using the second adjustment region.","Using this “two-stage” design, the result is that even when passing through the entire permissible temperature range only a maladjustment of the second adjustment region occurs which lies within permissible limits.","This can be achieved in that also when adjusting the second adjustment region an “over-adjustment” occurs, i.e., an adjustment such that the stresses frozen in the essential regions during heating and an associated flowing of the material in the essential regions cause in the first place a deformation of the actuator in a direction which leads to an “improvement” of the adjustment position.","According to an embodiment of the invention, the first adjustment region includes at least a double bridge.","This structure has the advantage of a significantly greater adjustment effect or setting range compared to a single bridge.","In a further development of the invention, the second adjustment region includes a single bridge.","In an embodiment of the invention, the first adjustment region (having the greater adjustment effect) can be provided lying closer to the support region of the actuator than the second adjustment region (having the lesser adjustment effect).","If angular adjustments of the actuator are carried out with the adjustment regions, i.e., the actuator is bent by an adjustment procedure (and not only shortened in the direction of the respective bridge), then only a relatively small, undesirable offset in the lateral direction is produced during adjustment procedures with the first adjustment region, besides the desired angular adjustment, since these two adjustment effects are not present in a decoupled manner in an actuator of this sort.","In any case, the second adjustment region produces only relatively small lateral adjustment movements as a consequence of the smaller adjustment effect compared to the first adjustment region.","In the preferred embodiment, the first adjustment region is provided so that a plane passing through it, which extends preferably perpendicularly to the axis of the actuator, essentially coincides with a plane which forms the basis for the adjustment procedure of an element arranged and to be adjusted in the support region.","In this manner, a lateral offset is reduced to zero for the desired angular adjustment.","In a further embodiment, the actuator has an essentially hollow-cylindrical casing in which the first and second adjustment regions are formed, in which each adjustment region preferably includes three bridges distributed over the circumference.","Using an actuator of this sort, a tilting of the element to be adjusted which is, for example, held centrally in the casing or in a floor or cover wall can be performed about two axes (tilting movements about two axes) as well as an axial adjustment (through uniform shortening of all three bridges).","Moreover, a lateral adjustment can take place if the bridges are positioned so that the tilting movements produce sufficient movements of the element to be adjusted (on a circular track).","This is the case particularly if the bridges have a sufficient axial distance from the element to be adjusted.","In one embodiment, the actuator is manufactured from an essentially planar element which includes a central region emanating from which a plurality (preferably three or six) of arms extend radially outwards, the arms being bent essentially at right angles to the central region.","An actuator of this sort can replace a tube actuator, but it is substantially easier and more cost-effective to manufacture.","In the simplest case, three arms can be provided at preferably an angular spacing of 120°, in which a first and a second adjustment region is provided in each arm.","In another embodiment, six arms can be provided at an angular spacing of 60°, in which a first and a second adjustment region is provided in every second arm.","If all of the arms are bent in the same direction and the longitudinal sides of the arms are joined in the bent state, preferably welded or soldered such that they form an essentially hollow-cylindrical wall, a tube actuator results in practical terms.","An actuator which is sealed with regard to the essentially cylindrical wall can be manufactured by forming the bridges as webs having a greater thickness than the regions provided to the side of the webs, in which the thickness of the regions provided to the side of the webs is chosen compared to the thickness of the webs so that a sufficient adjustment effect is obtainable during an adjustment procedure.","The bridges or rather webs are thus delimited to the side not by breakouts or recesses but by a wall having a small thickness.","If adjustment regions are formed in the tube actuator manufactured from a planar part with six arms only in every second arm, then in the remaining arms, preferably on the same axial position as the adjustment regions, regions with thinner walls can be provided in order to enable easier and better defined deformation of the actuator and thus a defined adjustment procedure.","This measure involving provision of regions with thinner walls which do not significantly hinder a deformation of the actuator during an adjustment procedure (and are bent or folded in this process), instead of breakouts between regions of the actuator which should be offset or tilted with respect to one another, is generally applicable to every geometric structure of an actuator.","In this manner, actuators that are impermeable at least in certain regions, and particularly gas-impermeable actuators, can be manufactured.","As was mentioned above, a tube actuator can be manufactured with an impermeable wall.","If the front sides of the tube actuator are also closed, an element to be adjusted that is held in the tube actuator can be shielded against ambient influences.","Here, a tube actuator can be manufactured preferably from one or more planar elements.","With a planar design, thin wall parts can be easily manufactured, e.g., using stamping techniques.","For example, a tube actuator could also be manufactured, instead of using the previously described bending of arms of a corresponding planar part, from an essentially rectangular planar part, which is correspondingly bent or “rolled” for this purpose.","The joining of the edges can take place through welding or soldering.","In another embodiment, when using a starting element having a central region and six arms, in each case three arms can be bent in alternating order in each case in the opposite direction by essentially 90°.","An actuator of this sort can accommodate in the central region an element to be adjusted and also be joined on the ends of the three respective arms in each case to an element to be adjusted or rather a support element in which an element to be adjusted is held.","If in each case two adjustment regions are provided in each arm, then the three elements can be adjusted to one another.","Here, in the first place an adjustment of two elements can take place which are held in an end region of the three arms of one side of the actuator or rather in its central region.","In the next step, an adjustment of the element held in the end region of the other three arms can then take place.","An optical component according to the invention includes a support element and an actuator on which an optical element to be adjusted is arranged.","It can be adjusted using an actuator with respect to the support element or to a further optical element joined to it.","Within the meaning of the present invention, an optical element is understood to be any element which serves to guide or otherwise influence light, such as optical waveguides, lenses, filters, mirrors or the like.","In an embodiment, the support element can have an accommodation region for the essentially hollow-cylindrical wall of the actuator or the bent arms of the actuator, the accommodation region preferably including a stopping face for the front side of the hollow-cylindrical wall or the front sides of the bent arms.","Thus, the actuator can be placed in a defined axial position on the support element.","In this position, the two parts can be rigidly joined, for example, bonded, welded or soldered.","To mount and to adjust a further optical element, the support element can have a second accommodation region for a further actuator on whose support region a further optical element is provided, the further actuator including the first actuator and the optical element arranged on it.","Using an actuator which has six arms of which in each case three were bent in opposite directions by approx.","90°, in which on each of the arms a first and a second adjustment region is provided, an optical component can be implemented with two support elements, in which in each case one of the support elements is joined to in each case the three arms bent in one direction.","In each support element, an optical element can be held.","In the central region of the actuator, a further optical element can be held which can be adjusted relative to the support elements.","In this manner, for example, an optical coupler or passive multiplexer can be implemented.","In this case, in the support elements in each case one or more optical waveguides are held and in the central region one or more filters, mirrors or the like.","Further embodiments of the invention follow from the dependent claims.","The invention is described hereafter in greater detail based on the figures shown in the drawing.","The figures are as follows: FIG.","1: Schematic representations of the laser beam adjustment method; FIG.","2: An embodiment of a tube actuator according to the invention; FIG.","3: An embodiment of a three-arm actuator according to the invention; FIG.","4: A passive optical multiplexer according to the invention; and FIG.","5: An embodiment of a six-arm actuator according to the invention.","FIG.","2 shows a tube-shaped actuator 100 (tube actuator) for the laser beam adjustment method described in the introduction which includes a hollow-cylindrical casing or rather wall 102 and a cover 104.In the cover 104, a circular breakout 105 is provided which allows a light beam (not shown) to pass through in the longitudinal axis of the actuator 100.On the inner or outer side of the cover 104, an optical element to be adjusted, e.g., a lens, a mirror or reflection or transmission filter (not shown) can be arranged.","In the hollow-cylindrical wall 102, a first adjustment region 106 is provided which includes three double bridges 108 which in each case have an angular spacing of 120°.","In this manner, by shortening a double bridge 108 or rather by shortening a bridge 110, 112 of the double bridges 108, a tilting of the cover 104 can be achieved, the reason being that between the bridges 110, 112, breakthroughs 113 are provided so that upon shortening the bridges 110, 112 of a double bridge 108, the bridges 110, 112 of the two other double bridges 108 are stressed only on bending.","Using the double bridges 108, an element to be adjusted that is arranged on the cover can be tilted about two axes.","Simultaneously, the element to be adjusted is naturally also slightly displaced laterally (essentially parallel to the cover 104) since the tilting axes do not lie in the plane of the element to be adjusted.","However, the smaller the axial spacing of the double bridges 108 from the cover 104, the smaller the lateral offset upon tilting of the cover through the shortening of at least one of the bridges 110, 112 of the double bridges 108.Since the lateral and tilting movements do not take place in a decoupled manner, in the actuator according to FIG.","2 the double bridges 108 are backed up relatively close (in the axial direction) to the cover so that during an adjustment procedure the lateral offset of the element to be adjusted is negligibly small compared to the angular offset.","If in the actuator according to FIG.","2 in an adjustment procedure all of the double bridges 108 are shortened in the same manner, an axial adjustment of the element to be adjusted can also be achieved.","For example, the focus of a lens or the width of a laser beam can be axially adjusted.","After the first adjustment procedure using the double bridges 108, which can take place, for example, at room temperature, the individual bridges 110, 112 of the double bridges 108 are heated so that the frozen-in stresses are reduced.","Since the yield point of most materials decreases at higher temperatures, a heating of the bridges 108 (and of the surrounding regions in which stresses are frozen in) can be carried out up to the maximum temperature of the specification range so that during subsequent heating or cooling of the bridges 110, 112 to an arbitrary temperature within the specification range, no flowing of the material occurs and the actuator [does not become] maladjusted in these regions.","After this reduction of the stresses in the bridges 110, 112, which of course is associated with a maladjustment, a readjustment using the second adjustment region 114 can be carried out in the actuator according to FIG.","2.The second adjustment region 114 includes three simple bridges 116, which again are arranged in essentially the same plane (perpendicular to the axis of the actuator) at an angular spacing of 120°.","Between the bridges 116, breakthroughs 117 are provided in the casing 102.The principle of adjustment using the simple bridges 116 of the second adjustment region 114 corresponds to the principle described above in connection with the first adjustment region 106.During an adjustment of the simple bridges 116, they are not heated over the entire cross-section but rather only in a (preferably middle) subregion.","The subregions which are not heated beyond the yield point assume upon heating of the relevant region beyond the yield point the tensile stresses and after the cooling of this region the compressive strains.","Since the adjustment effect which is obtainable using a simple bridge 116 is substantially smaller than the adjustment region which is attainable using a double bridge, a finer adjustment can take place using the second adjustment region.","In addition, the effect of a maladjustment which took place during a subsequent heating of the essential regions of the second adjustment region to a higher temperature compared to the temperature at which the adjustment occurred is very small.","A repeated “tempering” in order to reduce the stresses of the simple bridge can thus be dispensed with.","Of course, it is still possible when adjusting the bridges 116 of the second adjustment region to carry out a certain “overadjustment”, i.e., to cause an excess of adjustment range for the element to be adjusted so that upon reduction of the stresses by means of a subsequent tempering process in the first place an “improvement” of the adjustment position of the element to be adjusted results.","FIG.","3 shows a three-arm actuator 200 which corresponds largely to the tube actuator 100 in terms of its function.","The actuator 200 is manufactured from a planar element 202, which can be manufactured, for example, as a stamped part.","The element 202 includes a central region 204 and three radially extending arms 206.The central region 204 has again a central circular breakthrough 208 which corresponds in terms of its function in the completed actuator 200 to the breakthrough 105 in the cover 104 of the actuator 100.To manufacture the actuator 200, following a stamping out of the element 202, the arms 206 are bent roughly at right angles so that the three-arm actuator 200 shown in FIG.","3b is created.","Each arm 206 has a double bridge 108 and a simple bridge 116 which correspond in terms of their functions largely to the functions of the corresponding elements which were described above in conjunction with the actuator 100 according to FIG.","2.Compared to the actuator according to FIG.","2, the actuator 200 according to FIG.","3 has the advantage that it is much easier to manufacture.","A modification of the actuator shown in FIG.","2, which is not shown, can be manufactured from a stamped part which has six arms with an angular spacing of 60° instead of three arms.","If the six arms are bent to one side and the lateral edges of the arms joined, e.g., soldered or welded, a tube actuator is produced according to FIG.","2.However, the distinction is that the actuator manufactured from the stamped part is substantially simpler and thus more cost-effective to manufacture.","FIG.","4 shows an optical component 300 which can be, for example, an optical coupler or a passive optical multiplexer.","The component 300 includes a first support element 302 in which two optical waveguides 304 are held.","The support element 302 has an accommodation region 306 for an actuator which in the embodiment according to FIG.","4 is a three-arm actuator 200 according to FIG.","3.The accommodation region 306 is a cylindrical prolongation on the front side of which the front sides of the optical waveguides 304 are accessible.","A shoulder on the rear end of the accommodation region 306 forms a stop 308 for the front-side ends of the arms of the actuator 200.Thus, the actuator 200 can be placed in a defined axial position on the support element 302.Then, the actuator can be rigidly joined to the support element, for example, bonded, soldered or welded.","On the inner side of the central region of the actuator 200, an optical element 310 to be adjusted, e.g., a filter or a partially transparent mirror, is arranged.","Using the actuator 200, the optical element 310 can be adjusted in terms of its angular position or rather its axial position with respect to the front surfaces of the optical waveguides 304.The support element 302 has, moreover, a further accommodation region 312 for a further actuator not shown in greater detail.","It can also be a three-arm actuator 200 according to FIG.","3 which is fastened on the support element 302 as was described previously for the first actuator.","The further actuator is used to hold and adjust a further support part 314 in which two optical waveguides 304 are also held.","The support element 314 can be formed identically to the first support element 302.The first holding region 306 can be fastened in the centric region of the further actuator, which is not shown.","In the place of the two actuators which are used in the optical element according to FIG.","4, a single actuator 400 according to FIG.","5 can also be used.","It is manufactured from a stamped part having six arms 402 which are bent alternately upwards or downwards.","Each of the arms 402 has a double bridge 106 and a single bridge 114 whose function was described previously.","In each case three arms on each side of the actuator 400 are joined to in each case a support element 302, 314.In the central region of the actuator 400, the optical element to be adjusted can be held.","The adjustment then takes place in the previously described manner.","Of course, one of the two support elements can also be joined in the first place to the actuator and then the element held in the central region of the actuator can be adjusted with respect to the front surfaces of the relevant optical waveguides 304.Then, the second support element can be joined to the remaining three arms of the actuator and the second support element adjusted with respect to the already adjusted parts."]],"string":"[\n [\n \"Adjustment Method, Particularly a Laser Adjustment Method, and an Actuator Suitable For the Same\",\n \"The invention relates to an adjustment method, especially for adjusting optical or fibre optical components.\",\n \"Said method involves locally heating, in a defined manner, at least one first partial region of an adjustment region (106, 114) of an actuator (100, 200, 400), in such a way that compressive strains appear in at least one second partial region of the adjustment region of the actuator (100, 200, 400) as a result of the fact that said second partial region prevents the thermal expansion of the heated partial area.\",\n \"When the yielding point σF of the material of the partial region is achieved, said compressive strains lead to a plastic deformation of the heated partial region.\",\n \"The heated first partial region shrinks during the cooling process, triggering a defined geometry modification of the actuator (100, 200, 400) following the cooling process.\",\n \"Due to the fact that the at least one second partial region prevents the shrinking, tensile stresses appear in the previously heated first partial region and the compressive strains are frozen in the at least one second region.\",\n \"According to the invention, the regions of the actuator (100, 200, 300, 400), in which the tensile stresses or compressive strains are frozen following the cooling process, are brought to a critical temperature (Tk) in relation to the operational temperature range of the actuator, after the cooling process, at least until the flow processes of the material at said critical temperature are largely completed.\",\n \"A second adjustment process is then carried out.\",\n \"The invention also relates to an actuator which is especially suitable for carrying out the inventive method, and an optical component comprising one such actuator.\"\n ],\n [\n \"1-22.\",\n \"(canceled) 23.A method for adjusting optical or fiber optic components, comprising the acts of: performing a first adjustment procedure, including: locally heating a selected subregion of an actuator such that compressive stresses arise in the selected subregion, which, upon reaching a yield point of the material of the selected subregion, results in plastic deformation of the selected subregion; cooling the selected subregion, such that the selected subregion is shortened, resulting in a defined geometric change of the actuator, wherein tensile stresses arise in a first subregion of the actuator and, in a second region of the actuator, the compressive stresses are frozen; and reheating the selected subregion to a specified temperature such that stresses in a portion thereof are relieved; and performing a second adjustment procedure.\",\n \"24.The method of claim 23, wherein second adjustment procedure comprises locally heating and cooling another subregion of the actuator, wherein the other subregion is positioned, such that, in comparison to the selected subregion, a smaller adjustment effect is caused if the same amount of energy is supplied.\",\n \"25.The method of claim 23, wherein, after the first adjustment procedure, the selected subregion is brought to a critical temperature with regard to the usage temperature range of the actuator, at least until flow processes of the material are substantially concluded at the critical temperature.\",\n \"26.The method of claim 25, wherein the critical temperature is selected to be essentially the temperature within the usage temperature range at which a minimum yield point of the material is attained within the usage temperature range.\",\n \"27.The method of claim 25, wherein the material of the selected subregion has a yield point that drops monotonically with increasing temperatures within the usage temperature range 28.The method of claim 27, wherein the critical temperature is selected to be essentially the temperature corresponding to an upper limit of the usage temperature range.\",\n \"29.The method of claim 23, wherein locally heating the selected subregion is performed using an Nd:YAG laser.\",\n \"30.The method of claim 23, wherein locally heating the selected subregion is performed using a diode laser.\",\n \"31.An actuator for adjusting an optical or a fiber optics component, comprising: a support region for accommodating the component to be adjusted; a first adjustment region having a double bridge; and a second adjustment region having a simple bridge, wherein the first adjustment region produces a greater adjustment effect when an adjustment procedure is performed using the first adjustment region than an adjustment effect produced when an adjustment procedure is performed using the second adjustment region.\",\n \"32.The actuator of claim 31, further comprising means for heating a first bridge of the double bridge, wherein thermal expansion of the first bridge of the double bridge is constrained by the second bridge of the double bridge.\",\n \"33.The actuator of claim 32, further comprising means for heating a first subregion of the simple bridge, wherein a second subregion of the simple bridge constrains thermal expansion of the first subregion of the simple bridge.\",\n \"34.The actuator of claim 31, further comprising a substantially cylindrical body having an end at which the support region is positioned.\",\n \"35.The actuator of claim 34, wherein the first adjustment region and the second adjustment region are positioned in the cylindrical body.\",\n \"36.The actuator of claim 35, wherein the first adjustment region is positioned to be closer to the support region than is the second adjustment region.\",\n \"37.The actuator of claim 35, wherein first adjustment region defines a plane that is substantially perpendicular to an axis of the cylindrical body and substantially coincides with a plane that forms the basis of an adjustment procedure that can be performed using the first adjustment region.\",\n \"38.The actuator of claim 35 wherein: the first adjustment region includes three double bridges positioned about a circumference of the cylindrical body; and the second adjustment region includes three simple bridges positioned about the circumference of the cylindrical body.\",\n \"39.A method of manufacturing an actuator for use with an optical or a fiber optics component, comprising: obtaining a substantially planar element having: a central region; and a plurality of at least three arms that are connected to and extend radially away from the central region; and bending the arms to form substantially right angles with respect to the central region, the bent arms defining a first adjustment region that defines a plane substantially parallel to the central region and a second adjustment region substantially parallel to the central region.\",\n \"40.The method of claim 39, wherein the plurality of at least three arms comprises exactly three arms at an angular spacing of 120°.\",\n \"41.The method of claim 40, wherein each of the plurality of at least three arms includes a double bridge that defines the first adjustment region and a simple bridge that defines the second adjustment region.\",\n \"42.The method of claim 39, wherein the plurality of at least three arms comprises exactly six arms at an angular spacing of 60°.\",\n \"43.The method of claim 42, wherein bending the arms comprises: bending the six arms in the same direction; and connecting adjacent the longitudinal sides of the six arms to form a hollow-cylindrical wall of the actuator.\",\n \"44.The method of claim 43, wherein alternating arms of the six arms include a double bridge that defines the first adjustment region and a simple bridge that defines the second adjustment region.\"\n ],\n [\n \"The invention concerns an adjustment method, particularly for adjusting optical or fiber optical components, with the features of the preamble of patent claim 1.Moreover, the invention concerns an actuator which is suitable for the same and an optical component.\",\n \"Laser adjustment methods and actuators suitable for the same have been developed in recent times to enable extremely precise adjustment, e.g., of micromechanical components.\",\n \"The basic principle of laser adjustment consists of heating a predetermined region of an actuator using a high-energy, preferably pulsed laser beam in a short time, in which the thermal expansion of the relevant region is blocked by corresponding further regions of the actuator.\",\n \"In this manner, compression strains build up in this heated region, which result in a plastic deformation of this region upon reaching the yield point.\",\n \"When this region is cooled down after the high-energy laser beam is switched off, the thermal shrinkage of this region is again essentially prevented by the further regions of the actuator.\",\n \"This leads to the build-up of tensile stresses in the previously heated region which lead to a defined deformation of the actuator, in which the adjustment of a component joined to the actuator is enabled in this process.\",\n \"The basic principle of such an adjustment method is illustrated in FIG.\",\n \"1.FIG.\",\n \"1a shows the starting state of a bar 3 made of a suitable material which is restrained between two rigid demarcations 1.At the starting temperature, e.g., room temperature (20° C.), the bar 3 has a length l0.The center region 3a of the bar 3 is heated by a laser beam 5 in a short time.\",\n \"This results in the first place in compressive strains σ(−) in the bar 3 since the thermal longitudinal expansion of the bar 3 is blocked by the rigid demarcations 1.In this phase, a negative expansion ε is also customarily defined which corresponds to the compressive strains formed in the bar 3.This phase of the adjustment method is illustrated in FIG.\",\n \"1b.\",\n \"If the compressive strains σ(−) exceed the temperature-dependent yield point σF, a plastic deformation of the bar 3 arises in the region 3a.\",\n \"Correspondingly, the compressive strains in the bar 3 are reduced.\",\n \"This situation is illustrated in FIG.\",\n \"1c.\",\n \"After the laser beam 5 is switched off, the region 3a of the bar 3 begins to cool off, causing a thermal shrinkage of the bar 3.In this process, tensile stresses σ(+) arise in the bar 3 which in actual practice frequently lie in the vicinity of the temperature-dependent yield point σF(T).\",\n \"This situation is represented symbolically in FIG.\",\n \"1d because seen in the longitudinal direction of the bar 3 whose ends are joined by means of spring elements 7 to the rigid demarcations 1 .\",\n \".\",\n \".\",\n \".\",\n \"[Note: incomplete sentence in the German original].\",\n \"The spring forces corresponding to the tensile stresses σ(+) cause a defined deformation of the actuator in a practically realized actuator.\",\n \"In this connection, FIG.\",\n \"1d also shows that the tensile stresses σ(+) arise through the shortening of the bar 3 due to the plastic deformation in the region 3a caused during the heating, in which the length of the bar l1 after the adjustment procedure at the starting temperature is smaller than the original length l0 at the starting temperature.\",\n \"A problem in the previously known adjustment methods consists in that, as was previously mentioned, the tensile stresses frozen in the bar 3 lie relatively close to the yield point σF.\",\n \"The same can apply also to the compressive strains which occur in those regions which block the thermal expansion or rather the thermal shrinkage of the bar.\",\n \"Since micromechanical or optical components or rather subassemblies in actual practice are always specified for a certain temperature range, e.g., a range from −40° C. to +80° C., and must fulfill predetermined requirements for accuracy and long-term stability within the specified range, there results in previously known adjustment methods a maladjustment of the actuator if the adjusted components or rather the subassembly is brought to a temperature in the upper region of the specified range and the original adjustment was carried out at a significantly lower temperature, such as room temperature.\",\n \"This effect is caused by the temperature dependency of the yield point σF, most materials which are suitable for the manufacture of actuators for laser adjustment methods having a yield point which decreases with increasing temperature.\",\n \"If a temperature is reached at which the yield point σF falls below the value of the frozen-in tensile stresses, this results in a flowing of the material and in a reduction of the tensile stresses to the value of the yield point σF at the relevant temperature.\",\n \"Naturally, this is associated with a corresponding maladjustment of the actuator which is not acceptable at least for components requiring extremely precise adjustment which must be specified over a wide temperature range.\",\n \"The underlying object of the invention is therefore to create an adjustment method, particularly for adjusting optical or fiber optical components, with which improved long-term stability of an adjusted actuator or rather a subassembly having such an actuator can be ensured within a predetermined temperature range, it being possible to carry out the method quickly and at a low cost.\",\n \"Moreover, the underlying object of the invention is to create a special actuator for simply carrying out the method and an optical component having an actuator of this sort.\",\n \"This objective is solved by the invention with the features of patent claim 1, 6 or 17.The invention is based on the insight that the adjustment of an actuator can be carried out in an advantageous manner in the first place in a first adjustment procedure at an arbitrary temperature, e.g., room temperature, that subsequently at least the essential regions of the actuator (in which stresses are frozen in as a consequence of the first adjustment procedure) can have their tension relieved by heating to a specified temperature, and that subsequently a readjustment of the actuator can be carried out again at an arbitrary temperature, preferably room temperature.\",\n \"In an embodiment of the method, as regions essential for relieving the tension, all regions in which, after the first adjustment procedure compressive strains or tensile stresses are frozen in, are brought to a critical temperature (Tk) with regard to the usage temperature range of the actuator, at least until flow processes of the material are largely concluded at the critical temperature.\",\n \"In this connection, a critical temperature (Tk) is understood to be a temperature at which the yield point has a value such that when passing through the entire usage temperature range, only minor maladjustments of the adjusted actuator can occur which lie within specified tolerances.\",\n \"With conventional actuator materials which have within a specification range a yield point which decreases at higher temperatures, one will choose the critical temperature in the upper region of the specification range or even a temperature lying above its upper limit To.\",\n \"According to the preferred embodiment, the second adjustment procedure is carried out by heating and cooling a first subregion of a further adjustment region of the actuator, the further adjustment region being constituted such that in comparison to the first adjustment region, a smaller adjustment effect would be caused if the same amount of energy is supplied.\",\n \"Naturally, the energy of the second adjustment procedure or rather the curve vs. time of the power for heating the relevant region can differ in the second adjustment procedure and can be adapted particularly to the desired adjustment effect or rather adjustment range.\",\n \"According to an embodiment of the invention, the critical temperature Tk can essentially be chosen to be that temperature within the usage temperature range at which the minimum yield point σF of the material is attained within the usage temperature range.\",\n \"In this case, it is ensured that within the usage temperature range, no flowing of the material occurs in those regions of the actuator in which internal stresses occur as a result of the adjustment procedure.\",\n \"The possibility explained above of choosing an even higher temperature assumes in actual practice that a corresponding heating of the essential regions is carried out only locally.\",\n \"This applies at least if the actuator is part of a subassembly in which components are also contained which may be subjected as a maximum temperature to the upper temperature T0 of the planned usage temperature range.\",\n \"The essential regions include those regions which are subject during an adjustment procedure to thermal shrinkage and also those regions which prevent such shrinkage.\",\n \"In another embodiment of the method, essentially only those regions can be heated to a specified stress-relieving temperature as essential regions in which compressive strains are frozen in after the first adjustment procedure, in which the stress-relieving temperature is chosen such that a thermal expansion of the relevant regions [is] produced by the heating [and] the tensile stresses in those regions in which tensile stresses are frozen in following the first adjustment procedure, are increased above the yield point so that flowing of the material occurs in these regions.\",\n \"In other words, through the heating of the regions with compressive strains and the resultant thermal expansion of these regions, a stretching of the regions with tensile stresses is carried out.\",\n \"The stress-relieving temperature is chosen preferably such that after the tension is relieved during a reheating of all regions in which stresses are frozen in to a critical temperature (cf., in this connection the explanations above), no flow processes and thus no maladjustment of the actuator occurs.\",\n \"The heating for the actual adjustment procedure and the heating for the heating after the adjustment procedure can take place, for example, using a laser, preferably an Nd:YAG laser or diode laser.\",\n \"A special actuator with which the method according to the invention can be implemented in a particularly easy manner has a support region for accommodating an element to be adjusted and has at least a first and a second adjustment region, in which the first adjustment region is formed so that when carrying out an adjustment procedure using the first adjustment region, a greater adjustment effect can be obtained than when carrying out a further adjustment procedure using the second adjustment region.\",\n \"Using this “two-stage” design, the result is that even when passing through the entire permissible temperature range only a maladjustment of the second adjustment region occurs which lies within permissible limits.\",\n \"This can be achieved in that also when adjusting the second adjustment region an “over-adjustment” occurs, i.e., an adjustment such that the stresses frozen in the essential regions during heating and an associated flowing of the material in the essential regions cause in the first place a deformation of the actuator in a direction which leads to an “improvement” of the adjustment position.\",\n \"According to an embodiment of the invention, the first adjustment region includes at least a double bridge.\",\n \"This structure has the advantage of a significantly greater adjustment effect or setting range compared to a single bridge.\",\n \"In a further development of the invention, the second adjustment region includes a single bridge.\",\n \"In an embodiment of the invention, the first adjustment region (having the greater adjustment effect) can be provided lying closer to the support region of the actuator than the second adjustment region (having the lesser adjustment effect).\",\n \"If angular adjustments of the actuator are carried out with the adjustment regions, i.e., the actuator is bent by an adjustment procedure (and not only shortened in the direction of the respective bridge), then only a relatively small, undesirable offset in the lateral direction is produced during adjustment procedures with the first adjustment region, besides the desired angular adjustment, since these two adjustment effects are not present in a decoupled manner in an actuator of this sort.\",\n \"In any case, the second adjustment region produces only relatively small lateral adjustment movements as a consequence of the smaller adjustment effect compared to the first adjustment region.\",\n \"In the preferred embodiment, the first adjustment region is provided so that a plane passing through it, which extends preferably perpendicularly to the axis of the actuator, essentially coincides with a plane which forms the basis for the adjustment procedure of an element arranged and to be adjusted in the support region.\",\n \"In this manner, a lateral offset is reduced to zero for the desired angular adjustment.\",\n \"In a further embodiment, the actuator has an essentially hollow-cylindrical casing in which the first and second adjustment regions are formed, in which each adjustment region preferably includes three bridges distributed over the circumference.\",\n \"Using an actuator of this sort, a tilting of the element to be adjusted which is, for example, held centrally in the casing or in a floor or cover wall can be performed about two axes (tilting movements about two axes) as well as an axial adjustment (through uniform shortening of all three bridges).\",\n \"Moreover, a lateral adjustment can take place if the bridges are positioned so that the tilting movements produce sufficient movements of the element to be adjusted (on a circular track).\",\n \"This is the case particularly if the bridges have a sufficient axial distance from the element to be adjusted.\",\n \"In one embodiment, the actuator is manufactured from an essentially planar element which includes a central region emanating from which a plurality (preferably three or six) of arms extend radially outwards, the arms being bent essentially at right angles to the central region.\",\n \"An actuator of this sort can replace a tube actuator, but it is substantially easier and more cost-effective to manufacture.\",\n \"In the simplest case, three arms can be provided at preferably an angular spacing of 120°, in which a first and a second adjustment region is provided in each arm.\",\n \"In another embodiment, six arms can be provided at an angular spacing of 60°, in which a first and a second adjustment region is provided in every second arm.\",\n \"If all of the arms are bent in the same direction and the longitudinal sides of the arms are joined in the bent state, preferably welded or soldered such that they form an essentially hollow-cylindrical wall, a tube actuator results in practical terms.\",\n \"An actuator which is sealed with regard to the essentially cylindrical wall can be manufactured by forming the bridges as webs having a greater thickness than the regions provided to the side of the webs, in which the thickness of the regions provided to the side of the webs is chosen compared to the thickness of the webs so that a sufficient adjustment effect is obtainable during an adjustment procedure.\",\n \"The bridges or rather webs are thus delimited to the side not by breakouts or recesses but by a wall having a small thickness.\",\n \"If adjustment regions are formed in the tube actuator manufactured from a planar part with six arms only in every second arm, then in the remaining arms, preferably on the same axial position as the adjustment regions, regions with thinner walls can be provided in order to enable easier and better defined deformation of the actuator and thus a defined adjustment procedure.\",\n \"This measure involving provision of regions with thinner walls which do not significantly hinder a deformation of the actuator during an adjustment procedure (and are bent or folded in this process), instead of breakouts between regions of the actuator which should be offset or tilted with respect to one another, is generally applicable to every geometric structure of an actuator.\",\n \"In this manner, actuators that are impermeable at least in certain regions, and particularly gas-impermeable actuators, can be manufactured.\",\n \"As was mentioned above, a tube actuator can be manufactured with an impermeable wall.\",\n \"If the front sides of the tube actuator are also closed, an element to be adjusted that is held in the tube actuator can be shielded against ambient influences.\",\n \"Here, a tube actuator can be manufactured preferably from one or more planar elements.\",\n \"With a planar design, thin wall parts can be easily manufactured, e.g., using stamping techniques.\",\n \"For example, a tube actuator could also be manufactured, instead of using the previously described bending of arms of a corresponding planar part, from an essentially rectangular planar part, which is correspondingly bent or “rolled” for this purpose.\",\n \"The joining of the edges can take place through welding or soldering.\",\n \"In another embodiment, when using a starting element having a central region and six arms, in each case three arms can be bent in alternating order in each case in the opposite direction by essentially 90°.\",\n \"An actuator of this sort can accommodate in the central region an element to be adjusted and also be joined on the ends of the three respective arms in each case to an element to be adjusted or rather a support element in which an element to be adjusted is held.\",\n \"If in each case two adjustment regions are provided in each arm, then the three elements can be adjusted to one another.\",\n \"Here, in the first place an adjustment of two elements can take place which are held in an end region of the three arms of one side of the actuator or rather in its central region.\",\n \"In the next step, an adjustment of the element held in the end region of the other three arms can then take place.\",\n \"An optical component according to the invention includes a support element and an actuator on which an optical element to be adjusted is arranged.\",\n \"It can be adjusted using an actuator with respect to the support element or to a further optical element joined to it.\",\n \"Within the meaning of the present invention, an optical element is understood to be any element which serves to guide or otherwise influence light, such as optical waveguides, lenses, filters, mirrors or the like.\",\n \"In an embodiment, the support element can have an accommodation region for the essentially hollow-cylindrical wall of the actuator or the bent arms of the actuator, the accommodation region preferably including a stopping face for the front side of the hollow-cylindrical wall or the front sides of the bent arms.\",\n \"Thus, the actuator can be placed in a defined axial position on the support element.\",\n \"In this position, the two parts can be rigidly joined, for example, bonded, welded or soldered.\",\n \"To mount and to adjust a further optical element, the support element can have a second accommodation region for a further actuator on whose support region a further optical element is provided, the further actuator including the first actuator and the optical element arranged on it.\",\n \"Using an actuator which has six arms of which in each case three were bent in opposite directions by approx.\",\n \"90°, in which on each of the arms a first and a second adjustment region is provided, an optical component can be implemented with two support elements, in which in each case one of the support elements is joined to in each case the three arms bent in one direction.\",\n \"In each support element, an optical element can be held.\",\n \"In the central region of the actuator, a further optical element can be held which can be adjusted relative to the support elements.\",\n \"In this manner, for example, an optical coupler or passive multiplexer can be implemented.\",\n \"In this case, in the support elements in each case one or more optical waveguides are held and in the central region one or more filters, mirrors or the like.\",\n \"Further embodiments of the invention follow from the dependent claims.\",\n \"The invention is described hereafter in greater detail based on the figures shown in the drawing.\",\n \"The figures are as follows: FIG.\",\n \"1: Schematic representations of the laser beam adjustment method; FIG.\",\n \"2: An embodiment of a tube actuator according to the invention; FIG.\",\n \"3: An embodiment of a three-arm actuator according to the invention; FIG.\",\n \"4: A passive optical multiplexer according to the invention; and FIG.\",\n \"5: An embodiment of a six-arm actuator according to the invention.\",\n \"FIG.\",\n \"2 shows a tube-shaped actuator 100 (tube actuator) for the laser beam adjustment method described in the introduction which includes a hollow-cylindrical casing or rather wall 102 and a cover 104.In the cover 104, a circular breakout 105 is provided which allows a light beam (not shown) to pass through in the longitudinal axis of the actuator 100.On the inner or outer side of the cover 104, an optical element to be adjusted, e.g., a lens, a mirror or reflection or transmission filter (not shown) can be arranged.\",\n \"In the hollow-cylindrical wall 102, a first adjustment region 106 is provided which includes three double bridges 108 which in each case have an angular spacing of 120°.\",\n \"In this manner, by shortening a double bridge 108 or rather by shortening a bridge 110, 112 of the double bridges 108, a tilting of the cover 104 can be achieved, the reason being that between the bridges 110, 112, breakthroughs 113 are provided so that upon shortening the bridges 110, 112 of a double bridge 108, the bridges 110, 112 of the two other double bridges 108 are stressed only on bending.\",\n \"Using the double bridges 108, an element to be adjusted that is arranged on the cover can be tilted about two axes.\",\n \"Simultaneously, the element to be adjusted is naturally also slightly displaced laterally (essentially parallel to the cover 104) since the tilting axes do not lie in the plane of the element to be adjusted.\",\n \"However, the smaller the axial spacing of the double bridges 108 from the cover 104, the smaller the lateral offset upon tilting of the cover through the shortening of at least one of the bridges 110, 112 of the double bridges 108.Since the lateral and tilting movements do not take place in a decoupled manner, in the actuator according to FIG.\",\n \"2 the double bridges 108 are backed up relatively close (in the axial direction) to the cover so that during an adjustment procedure the lateral offset of the element to be adjusted is negligibly small compared to the angular offset.\",\n \"If in the actuator according to FIG.\",\n \"2 in an adjustment procedure all of the double bridges 108 are shortened in the same manner, an axial adjustment of the element to be adjusted can also be achieved.\",\n \"For example, the focus of a lens or the width of a laser beam can be axially adjusted.\",\n \"After the first adjustment procedure using the double bridges 108, which can take place, for example, at room temperature, the individual bridges 110, 112 of the double bridges 108 are heated so that the frozen-in stresses are reduced.\",\n \"Since the yield point of most materials decreases at higher temperatures, a heating of the bridges 108 (and of the surrounding regions in which stresses are frozen in) can be carried out up to the maximum temperature of the specification range so that during subsequent heating or cooling of the bridges 110, 112 to an arbitrary temperature within the specification range, no flowing of the material occurs and the actuator [does not become] maladjusted in these regions.\",\n \"After this reduction of the stresses in the bridges 110, 112, which of course is associated with a maladjustment, a readjustment using the second adjustment region 114 can be carried out in the actuator according to FIG.\",\n \"2.The second adjustment region 114 includes three simple bridges 116, which again are arranged in essentially the same plane (perpendicular to the axis of the actuator) at an angular spacing of 120°.\",\n \"Between the bridges 116, breakthroughs 117 are provided in the casing 102.The principle of adjustment using the simple bridges 116 of the second adjustment region 114 corresponds to the principle described above in connection with the first adjustment region 106.During an adjustment of the simple bridges 116, they are not heated over the entire cross-section but rather only in a (preferably middle) subregion.\",\n \"The subregions which are not heated beyond the yield point assume upon heating of the relevant region beyond the yield point the tensile stresses and after the cooling of this region the compressive strains.\",\n \"Since the adjustment effect which is obtainable using a simple bridge 116 is substantially smaller than the adjustment region which is attainable using a double bridge, a finer adjustment can take place using the second adjustment region.\",\n \"In addition, the effect of a maladjustment which took place during a subsequent heating of the essential regions of the second adjustment region to a higher temperature compared to the temperature at which the adjustment occurred is very small.\",\n \"A repeated “tempering” in order to reduce the stresses of the simple bridge can thus be dispensed with.\",\n \"Of course, it is still possible when adjusting the bridges 116 of the second adjustment region to carry out a certain “overadjustment”, i.e., to cause an excess of adjustment range for the element to be adjusted so that upon reduction of the stresses by means of a subsequent tempering process in the first place an “improvement” of the adjustment position of the element to be adjusted results.\",\n \"FIG.\",\n \"3 shows a three-arm actuator 200 which corresponds largely to the tube actuator 100 in terms of its function.\",\n \"The actuator 200 is manufactured from a planar element 202, which can be manufactured, for example, as a stamped part.\",\n \"The element 202 includes a central region 204 and three radially extending arms 206.The central region 204 has again a central circular breakthrough 208 which corresponds in terms of its function in the completed actuator 200 to the breakthrough 105 in the cover 104 of the actuator 100.To manufacture the actuator 200, following a stamping out of the element 202, the arms 206 are bent roughly at right angles so that the three-arm actuator 200 shown in FIG.\",\n \"3b is created.\",\n \"Each arm 206 has a double bridge 108 and a simple bridge 116 which correspond in terms of their functions largely to the functions of the corresponding elements which were described above in conjunction with the actuator 100 according to FIG.\",\n \"2.Compared to the actuator according to FIG.\",\n \"2, the actuator 200 according to FIG.\",\n \"3 has the advantage that it is much easier to manufacture.\",\n \"A modification of the actuator shown in FIG.\",\n \"2, which is not shown, can be manufactured from a stamped part which has six arms with an angular spacing of 60° instead of three arms.\",\n \"If the six arms are bent to one side and the lateral edges of the arms joined, e.g., soldered or welded, a tube actuator is produced according to FIG.\",\n \"2.However, the distinction is that the actuator manufactured from the stamped part is substantially simpler and thus more cost-effective to manufacture.\",\n \"FIG.\",\n \"4 shows an optical component 300 which can be, for example, an optical coupler or a passive optical multiplexer.\",\n \"The component 300 includes a first support element 302 in which two optical waveguides 304 are held.\",\n \"The support element 302 has an accommodation region 306 for an actuator which in the embodiment according to FIG.\",\n \"4 is a three-arm actuator 200 according to FIG.\",\n \"3.The accommodation region 306 is a cylindrical prolongation on the front side of which the front sides of the optical waveguides 304 are accessible.\",\n \"A shoulder on the rear end of the accommodation region 306 forms a stop 308 for the front-side ends of the arms of the actuator 200.Thus, the actuator 200 can be placed in a defined axial position on the support element 302.Then, the actuator can be rigidly joined to the support element, for example, bonded, soldered or welded.\",\n \"On the inner side of the central region of the actuator 200, an optical element 310 to be adjusted, e.g., a filter or a partially transparent mirror, is arranged.\",\n \"Using the actuator 200, the optical element 310 can be adjusted in terms of its angular position or rather its axial position with respect to the front surfaces of the optical waveguides 304.The support element 302 has, moreover, a further accommodation region 312 for a further actuator not shown in greater detail.\",\n \"It can also be a three-arm actuator 200 according to FIG.\",\n \"3 which is fastened on the support element 302 as was described previously for the first actuator.\",\n \"The further actuator is used to hold and adjust a further support part 314 in which two optical waveguides 304 are also held.\",\n \"The support element 314 can be formed identically to the first support element 302.The first holding region 306 can be fastened in the centric region of the further actuator, which is not shown.\",\n \"In the place of the two actuators which are used in the optical element according to FIG.\",\n \"4, a single actuator 400 according to FIG.\",\n \"5 can also be used.\",\n \"It is manufactured from a stamped part having six arms 402 which are bent alternately upwards or downwards.\",\n \"Each of the arms 402 has a double bridge 106 and a single bridge 114 whose function was described previously.\",\n \"In each case three arms on each side of the actuator 400 are joined to in each case a support element 302, 314.In the central region of the actuator 400, the optical element to be adjusted can be held.\",\n \"The adjustment then takes place in the previously described manner.\",\n \"Of course, one of the two support elements can also be joined in the first place to the actuator and then the element held in the central region of the actuator can be adjusted with respect to the front surfaces of the relevant optical waveguides 304.Then, the second support element can be joined to the remaining three arms of the actuator and the second support element adjusted with respect to the already adjusted parts.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473941"}}},{"rowIdx":769,"cells":{"text":{"kind":"list like","value":[["Bioluminescent plants and methods of making same","A genetically modified plant cell containing a heterologous nucleotide sequence encoding a bioluminescent polypeptide, which is expressed in amount sufficient to produce at least about 750,000 photons of visible light/mm2/second, is provided, as is a visibly bioluminescent transgenic plant, which contains such a genetically modified plant cell.","Also provided is a recombinant nucleic acid molecule, which contains a plant translational enhancer operatively linked to a nucleotide sequence encoding a bioluminescent polypeptide.","In addition, methods of producing a genetically modified plant cell that is visibly bioluminescent introducing a transgene encoding a bioluminescent polypeptide into a plant cell, whereby the bioluminescent polypeptide is expressed at a level that produces at least about 750,000 photons of visible light/mm2/second are provided, as are kits containing such visibly bioluminescent compositions."],["1.A genetically modified plant cell, comprising a heterologous nucleotide sequence encoding a bioluminescent polypeptide, wherein the bioluminescent polypeptide can be expressed in amount sufficient to produce at least 750,000 photons of visible light/mm2/second.","2.The genetically modified plant cell of claim 1, wherein the bioluminescent polypeptide is luciferase.","3.The genetically modified plant cell of claim 1, wherein the bioluminescent polypeptide is a luciferase variant.","4.The genetically modified plant cell of claim 3, wherein the luciferase variant is a luc+ luciferase variant.","5.The genetically modified plant cell of claim 4, wherein the luc+ luciferase variant is expressed in the plant cell at a level of at least 100 pg/μg protein as determined by western blot analysis.","6.The genetically modified plant cell of claim 1, wherein the bioluminescent polypeptide can be expressed in amount sufficient to produce at least one million photons of visible light/mm2/second.","7.The genetically modified plant cell of claim 1, which has a reduced level of chlorophyll as compared to corresponding wild type plant cell.","8.A transgenic plant, comprising the genetically modified plant cell of claim 1 or claim 7.9.A plant cell or tissue obtained from the transgenic plant of claim 8.10.A cutting of the transgenic plant of claim 8.11.A seed produced by the transgenic plant of claim 8.12.A cDNA or genomic DNA library prepared from the transgenic plant of claim 8, or from a plant cell or plant tissue obtained from said transgenic plant.","13.The transgenic plant of claim 8, wherein the plant is a monocot.","14.The transgenic plant of claim 8, wherein the plant is a dicot.","15.The transgenic plant of claim 14, wherein the plant is an angiosperm.","16.The transgenic plant of claim 15, wherein the angiosperm is a cereal plant, a leguminous plant, an oilseed plant, or a hardwood tree.","17.The transgenic plant of claim 8, wherein the plant is an ornamental plant.","18.The transgenic plant of claim 17, wherein the ornamental plant is a petunia.","19.The transgenic plant of claim 17, wherein the ornamental plant is a carnation.","20.The genetically modified plant of claim 1, wherein the heterologous nucleotide sequence encoding a bioluminescent polypeptide comprises nucleotides 8366 to 11,113 of SEQ ID NO:2.21.The genetically modified plant of claim 1, wherein the heterologous nucleotide sequence encoding a bioluminescent polypeptide comprises nucleotides 8340 to 12,098 of SEQ ID NO:4 or nucleotides 6 to 3570 of SEQ ID NO:5.22.A recombinant nucleic acid molecule, comprising, in operative linkage, a plant translational enhancer and a nucleotide sequence encoding a bioluminescent polypeptide.","23.The recombinant nucleic acid molecule of claim 22, wherein the plant translational enhancer is a plant potyvirus translational enhancer.","24.The recombinant nucleic acid molecule of claim 23, wherein the plant potyvirus is a tobacco etch virus (TEV) or an omega enhancer.","25.The recombinant nucleic acid molecule of claim 22, wherein the bioluminescent polypeptide is a luciferase polypeptide or variant thereof.","26.The recombinant nucleic acid molecule of claim 22, further comprising an operatively linked transcriptional regulatory element, which comprises a promoter.","27.The recombinant nucleic acid molecule of claim 26, wherein the transcriptional regulatory element is a cauliflower mosaic virus (CaMV) 35S regulatory element, a metallothionein regulatory element, or an actin 2 regulatory element.","28.The recombinant nucleic acid molecule of claim 22, comprising, in operative linkage, a CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and a CaMV 35S terminator.","29.The recombinant nucleic acid molecule of claim 28, wherein the CaMV 35S enhancer is a dual CaMV 35S enhancer.","30.The recombinant nucleic acid molecule of claim 28, wherein the luciferase polypeptide or variant thereof is a luc+ polypeptide.","31.The recombinant nucleic acid molecule of claim 22, comprising, in operative linkage, a metallothionein gene transcriptional regulatory element, an omega translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and an RbcS E9 polyA sequence.","32.The recombinant nucleic acid molecule of claim 22, comprising, in operative linkage, an actin 2 gene transcriptional regulatory element, an omega translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and an RbcS E9 polyA sequence.","33.The recombinant nucleic acid molecule of claim 22, comprising nucleotides 8366 to 11,113 of SEQ ID NO:2.34.The recombinant nucleic acid molecule of claim 22, comprising nucleotides 8340 to 12,098 of SEQ ID NO:4 or nucleotides 6 to 3570 of SEQ ID NO:5.35.A vector, comprising the recombinant nucleic acid molecule of claim 22.36.The vector of claim 32, wherein the vector is an expression vector.","37.The vector of claim 35, wherein the recombinant nucleic acid molecule comprises, in operative linkage, a CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and a CaMV 35S terminator.","38.The vector of claim 35, wherein the recombinant nucleic acid molecule comprises, in operative linkage, a metallothionein gene transcriptional regulatory element, an omega translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and an RbcS E9 polyA sequence.","39.The vector of claim 35, wherein the recombinant nucleic acid molecule comprises, in operative linkage, an actin 2 gene transcriptional regulatory element, an omega translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and an RbcS E9 polyA sequence.","40.The vector of claim 35, wherein the recombinant nucleic acid molecule comprises nucleotides 8366 to 11,113 of SEQ ID NO:2.41.The vector of claim 35, wherein the recombinant nucleic acid molecule comprises nucleotides 8340 to 12, 098 of SEQ ID NO:4 or nucleotides 6 to 3570 of SEQ ID NO:5.42.The vector of claim 35, which is selected from SEQ ID NO:1 and SEQ ID NO:2.43.The vector of claim 35, which is selected from SEQ ID NO:4 and SEQ ID NO:5.44.A cell containing the recombinant nucleic acid molecule of claim 22.45.A cell containing the vector of claim 35.46.A method of producing a genetically modified plant cell that is visibly bioluminescent, the method comprising introducing a transgene comprising a nucleotide sequence encoding a bioluminescent polypeptide into a plant cell, whereby the bioluminescent polypeptide is expressed at a level that produces at least 750,000 photons of visible light/mm2/second, thereby producing a genetically modified plant cell that visibly bioluminesce.","47.The method of claim 45, wherein the transgene further comprises a plant translational enhancer operatively linked to the nucleotide sequence encoding a bioluminescent polypeptide.","48.The method of claim 46, wherein the plant translational enhancer is a plant potyvirus translational enhancer selected from a tobacco etch virus (TEV) translational enhancer and a tobacco mosaic virus translational enhancer.","49.The method of claim 46, wherein the plant translational enhancer is an omega translational enhancer having a nucleotide sequence set forth as nucleotides 1169 to 1235 of SEQ ID NO:5.50.The method of claim 46, wherein the bioluminescent polypeptide is a luciferase polypeptide or variant thereof.","51.The method of claim 47, wherein the transgene further comprises a transcriptional regulatory element operatively linked to the plant translational enhancer and nucleotide sequence encoding a bioluminescent polypeptide.","52.The method of claim 51, wherein the transcriptional regulatory element is a cauliflower mosaic virus (CaMV) 35S regulatory element, a metallothionein gene regulatory element, or an actin gene regulatory element.","53.The method of claim 46, wherein the transgene comprises, in operative linkage, a dual CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and a CaMV 35S terminator.","54.The method of claim 53, wherein the luciferase polypeptide or variant thereof is a luc+ polypeptide.","55.The method of claim 46, wherein the transgene comprises, in -operative linkage, a metallothionein gene transcriptional regulatory element, which comprises a promoter and, optionally, an enhancer; an omega translational enhancer; a nucleotide sequence encoding a luciferase polypeptide or variant thereof; and an RbcS E9 polyA sequence.","56.The method of claim 55, wherein the luciferase polypeptide or variant thereof is a luc+ polypeptide.","57.The method of claim 46, wherein the transgene comprises, in operative linkage, an actin 2 gene transcriptional regulatory element, which comprises a promoter and, optionally, an enhancer; an omega translational enhancer; a nucleotide sequence encoding a luciferase polypeptide or variant thereof; and an RbcS E9 polyA sequence.","58.The method of claim 57, wherein the luciferase polypeptide or variant thereof is a luc+ polypeptide.","59.The method of claim 46, wherein the transgene comprises nucleotides 8366 to 11,113 of SEQ ID NO:2, nucleotides 8340 to 12,098 of SEQ ID NO:4 or nucleotides 6 to 3570 of SEQ ID NO:5.60.The method of claim 46, wherein the transgene is stably maintained in the plant cell genome.","61.The method of claim 60, wherein the transgene is integrated into the plant cell genome.","62.The method of claim 46, further comprising contacting the plant cell, or the genetically modified plant cell derived therefrom, with an agent that inhibits carotenoid synthesis, thereby reducing the amount of chlorophyll in the plant cell.","63.The method of claim 62, wherein the agent is norfluorazon.","64.A genetically modified plant cell produced by the method of claim 46 or claim 62.65.A transgenic plant comprising the genetically modified plant cell of claim 64.66.A genetically modified plant cell, comprising a heterologous nucleotide, which comprises, in operative linkage, a CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and a CaMV 35S terminator wherein the luciferase polypeptide or variant thereof can be expressed in amount sufficient to produce at least 750,000 photons of visible light/mm2/second.","67.The genetically modified plant cell of claim 66, wherein the heterologous nucleotide sequence encoding a bioluminescent polypeptide comprises nucleotides 8366 to 11,113 of SEQ ID NO:2.68.A transgenic plant, comprising the genetically modified plant cell of claim 66 or claim 67.69.A plant cell or tissue obtained from the transgenic plant of claim 68.70.A cutting of the transgenic plant of claim 68.71.A seed produced by the transgenic plant of claim 68.72.A cDNA or genomic DNA library prepared from the transgenic plant of claim 68, or from a plant cell or plant tissue obtained from said transgenic plant.","73.A genetically modified plant cell, comprising a heterologous nucleotide sequence, which comprises, in operative linkage, a metallothionein gene transcriptional regulatory element, an omega translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and an RbcS E9 polyA sequence., wherein the luciferase polypeptide or variant thereof can be expressed in amount sufficient to produce at least 750,000 photons of visible light/mm2/second.","74.The genetically modified plant cell of claim 73, wherein the heterologous nucleotide sequence comprises nucleotides 8340 to 12,098 of SEQ ID NO:4 75.A transgenic plant, comprising the genetically modified plant cell of claim 73 or claim 74.76.A plant cell or tissue obtained from the transgenic plant of claim 75.77.A cutting of the transgenic plant of claim 75.78.A seed produced by the transgenic plant of claim 75.79.A genetically modified plant cell, comprising a heterologous nucleotide sequence, which comprises, in operative linkage, an actin 2 gene transcriptional regulatory element, an omega translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and an RbcS E9 polyA sequence, wherein the luciferase polypeptide or variant thereof can be expressed in amount sufficient to produce at least 750,000 photons of visible light/mm2/second.","80.The genetically modified plant cell of claim 79, wherein the heterologous nucleotide sequence comprises nucleotides 6 to 3570 of SEQ ID NO:5.81.A transgenic plant, comprising the genetically modified plant cell of claim 79 or claim 80.82.A plant cell or tissue obtained from the transgenic plant of claim 81.83.A cutting of the transgenic plant of claim 81.84.A seed produced by the transgenic plant of claim 81.85.A kit, comprising the genetically modified plant cell of claim 1 or claim 7, or a derivative of the genetically modified plant cell.","86.A kit comprising a derivative of the genetically modified plant cell, wherein the derivative of the genetically modified plant cell is a transgenic plant comprising the genetically modified plant cell of claim 1 or claim 7, or a cell, tissue or organ of said transgenic plant.","87.The kit of claim 86, wherein the organ of said transgenic plant is a flower or a bract.","88.The kit of claim 85, further comprising an amount of luciferin sufficient for generating visible bioluminescence of the genetically modified plant cell or the derivative of the genetically modified plant cell.","89.The kit of claim 85, further comprising an amount of a carotenoid inhibitor sufficient for reducing or inhibiting chlorophyll production in the genetically modified plant cell or the derivative of the genetically modified plant cell.","90.A kit, comprising a cutting or a seed of the transgenic plant of claim 8.91.The kit of claim 90, further comprising reagents for growing a transgenic plant from the cutting or seed.","92.The kit of claim 90, further comprising an amount of a carotenoid inhibitor sufficient for reducing or inhibiting chlorophyll production in a transgenic plant grown from the cutting or seed.","93.The kit of claim 90, further comprising an amount of luciferin sufficient for generating visible bioluminescence of a transgenic plant grown from the cutting or seed.","94.A kit, comprising the genetically modified plant cell of claim 66, 67, 73, 74, 79, or 80, or a derivative of the genetically modified plant cell.","95.A kit comprising a derivative of a genetically modified plant cell, wherein the derivative of the genetically modified plant cell is a transgenic plant comprising the genetically modified plant cell of claim 66, 67, 73, 74, 79 or 80, or a cell, tissue or organ of said transgenic plant.","96.The kit of claim 95, wherein the organ of said transgenic plant is a flower or a bract.","97.The kit of claim 94, further comprising an amount of luciferin sufficient for generating visible bioluminescence of the genetically modified plant cell or the derivative of the genetically modified plant cell.","98.The kit of claim 94, further comprising an amount of a carotenoid inhibitor sufficient for reducing or inhibiting chlorophyll production in the genetically modified plant cell or the derivative of the genetically modified plant cell.","99.A kit, comprising a cutting of the transgenic plant of claim 68, or a seed produced by said transgenic plant.","100.The kit of claim 99, further comprising reagents for growing a transgenic plant from the cutting or seed.","101.The kit of claim 99, further comprising an amount of a carotenoid inhibitor sufficient for reducing or inhibiting chlorophyll production in a transgenic plant grown from the cutting or seed.","102.The kit of claim 99, further comprising an amount of luciferin sufficient for generating visible bioluminescence of a transgenic plant grown from the cutting or seed.","103.The kit of claim 95, further comprising an amount of luciferin sufficient for generating visible bioluminescence of the genetically modified plant cell or the derivative of the genetically modified plant cell.","104.The kit of claim 95, further comprising an amount of a carotenoid inhibitor sufficient for reducing or inhibiting chlorophyll production in the genetically modified plant cell or the derivative of the genetically modified plant cell.","105.A kit comprising a cutting of the transgenic plant of claim 75, or a seed produced by said transgenic plant.","106.The kit of claim 105, further comprising reagents for growing a transgenic plant from the cutting or seed.","107.The kit of claim 105, further comprising an amount of a carotenoid inhibitor sufficient for reducing or inhibiting chlorophyll production in a transgenic plant grown from the cutting or seed.","108.The kit of claim 105, further comprising an amount of luciferin sufficient for generating visible bioluminescence of a transgenic plant grown from the cutting or seed.","109.A kit comprising a cutting of the transgenic plant of claim 81, or a seed produced by said transgenic plant.","110.The kit of claim 109, further comprising reagents for growing a transgenic plant from the cutting or seed.","111.The kit of claim 110 further comprising an amount of a carotenoid inhibitor sufficient for reducing or inhibiting chlorophyll production in a transgenic plant grown from the cutting or seed.","112.The kit of claim 109, further comprising an amount of luciferin sufficient for generating visible bioluminescence of a transgenic plant grown from the cutting or seed."],["<SOH> BACKGROUND OF THE INVENTION <EOH>"],["<SOH> SUMMARY OF THE INVENTION <EOH>The present invention relates to a genetically modified plant cell that exhibits bioluminescence that is visible to the naked eye.","Such a genetically modified plant cell contains a heterologous nucleotide sequence that encodes a bioluminescent polypeptide, which, upon expression, can result in the production of at least about 750,000 photons of visible light/mm 2 /second by the plant cell.","The bioluminescent polypeptide can be any polypeptide that is bioluminescent, or can act in concert with a second molecule such that visible light is produced.","In one embodiment, the genetically modified plant cell contains a heterologous nucleotide sequence encoding luciferase or a luciferase variant, for example, luc + .","Upon contact of a genetically modified plant expressing luciferase or a variant thereof with luciferin, a sufficient amount of visible light is produced such that the light is visible to the naked eye.","Such a genetically modified plant cell generally expresses at least about 100 pg luc + /μg protein as determined by western blot analysis such that, upon contact of the plant cell with luciferin, the cell is visibly bioluminescent.","In another embodiment, the genetically modified plant cell, or the plant cell from which the genetically modified plant cell is derived, is contacted with an agent that reduces or inhibits pigment formation in the plant, for example, carotenoid synthesis, thereby reducing or inhibiting chlorophyll synthesis.","Since chlorophyll in a plant cell can absorb photons generated by the luciferase in the genetically modified plant cell, contact of the genetically modified plant cell, or a transgenic plant derived therefrom, with an amount of a carotenoid inhibitor sufficient to reduce or inhibit chlorophyll synthesis provides a means to enhance photon emission from the plant cell and, therefore, enhance the visible bioluminescence.","In still another embodiment, a visibly bioluminescent transgenic plant is generated from the genetically modified plant cell containing a heterologous nucleotide sequence encoding a bioluminescent polypeptide.","As such, the invention relates to such a transgenic plant, as well as to a plant cell or tissue obtained from the transgenic plant, a seed produced by the transgenic plant, and a cDNA or genomic DNA library prepared from the transgenic plant or from a plant cell or plant tissue obtained from the transgenic plant.","The transgenic plant can be a monocot, or can be a dicot, for example, an angiosperm such as a cereal plant, a leguminous plant, an oilseed plant, a hardwood tree, or an ornamental plant such as a petunia, an orchid, a carnation or the like.","The present invention also relates to a recombinant nucleic acid molecule, which comprises a translational enhancer, which enhances translation in a plant cell, operatively linked to a nucleotide sequence encoding a bioluminescent polypeptide.","The translational enhancer can be any translational enhancer, for example, a plant potyvirus translational enhancer such as tobacco etch virus (TEV) translational enhancer, a tobacco mosaic virus omega translational enhancer, or a translational enhancer comprising a Kozak sequence.","The encoded bioluminescent polypeptide can be a luciferase polypeptide or variant thereof, for example, the luc + luciferase variant.","A recombinant nucleic acid molecule of the invention also can contain a transcriptional regulatory element, i.e., promoter, enhancer, etc., that enhances transcription in a plant cell, for example, a plant virus enhancer such as a cauliflower mosaic virus (CaMV) 35S enhancer, or a regulatory element from any other organism, for example, an actin 2 regulatory element or a metallothionein regulatory element, which comprises a promoter and, optionally, an enhancer, wherein the transcriptional regulatory element is operatively linked to the plant translational enhancer and nucleotide sequence encoding the bioluminescent polypeptide.","In one embodiment, a recombinant nucleic acid molecule contains, in operative linkage, a CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and a CaMV 35S terminator.","In another embodiment, the CaMV 35S enhancer is a dual CaMV 35S enhancer, and the luciferase polypeptide or variant thereof is the luc + luciferase variant.","Such a recombinant nucleic acid molecule is exemplified herein by the nucleotide sequence set forth as nucleotides 8366 to 11,113 of SEQ ID NO:2.In still other embodiments, the transcriptional regulatory element in a recombinant nucleic molecule of the invention is a metallothionein regulatory element or an actin 2 regulatory element, and the translational enhancer is an omega enhancer, for example, in a recombinant nucleic acid molecule having a sequence set forth as nucleotides 8340 to 12,098 of SEQ ID NO:4 or as nucleotides 6 to 3570 of SEQ ID NO:5.The present invention further relates to a vector containing a recombinant nucleic acid molecule comprising a translational enhancer active in a plant operatively linked to a nucleotide sequence encoding a bioluminescent polypeptide.","Preferably, the vector is an expression vector.","In one embodiment, the recombinant nucleic acid molecule contained in the vector comprises, in operative linkage, a CAMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and a CaMV 35S terminator.","In another embodiment, the recombinant nucleic acid molecule contained in the vector comprises, in operative linkage, a dual CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luc + luciferase variant, and a CaMV 35S terminator.","In still another embodiment, the recombinant nucleic acid molecule contained in the vector comprises the nucleotide sequence set forth as nucleotides 8366 to 11,113 of SEQ ID NO:2.In yet another embodiment, the transcriptional regulatory element in the recombinant nucleic acid molecule comprises an actin 2 promoter or a metallothionein promoter, and the translation enhancer is an omega enhancer, for example, in the recombinant nucleic acid molecules set forth as nucleotides 8340 to 12,098 of SEQ D) NO:4 or as nucleotides 6 to 3570 of SEQ ID NO:5.Vectors of the invention are exemplified herein by those as set forth in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:4 and SEQ ID NO:5.The present invention also provides a cell containing a recombinant nucleic acid molecule of the invention, or containing a vector of the invention.","The present invention further relates to a method of producing a genetically modified plant cell that is visibly bioluminescent.","A method of the invention can be performed, for example, by introducing a transgene comprising a nucleotide sequence encoding a bioluminescent polypeptide into a plant cell, whereby the bioluminescent polypeptide is expressed at a level that produces at least about 750,000 photons of visible light/mm 2 /second.","The transgene useful in a method of making a genetically modified plant cell that is visibly bioluminescent generally includes a translational enhancer that is active in a plant, for example, a plant translational enhancer such as a plant potyvirus translational enhancer (e.g., a TEV translational enhancer), a tobacco mosaic virus (TMV) translational enhancer such as the omega enhancer, operatively linked to the nucleotide sequence encoding a bioluminescent polypeptide, for example, a luciferase polypeptide or variant thereof.","The transgene also can contain a plant virus enhancer, for example, a CaMV 35S enhancer, which is operatively linked to the plant translational enhancer and nucleotide sequence encoding a bioluminescent polypeptide.","In one embodiment, a transgene useful in making a visibly luminescent genetically modified plant cell contains, in operative linkage, a dual CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and a CaMV 35S terminator.","In another embodiment, the nucleotide sequence encodes a luc + luciferase variant polypeptide.","Such transgenes are exemplified by the nucleotide sequences set forth as nucleotides 8366 to 11,113 of SEQ ID NO:2, nucleotides 8340 to 12,098 of SEQ ID NO:4, and nucleotides 6 to 3570 of SEQ ID NO:5.A method of the invention can further include contacting the plant cell, or the genetically modified plant cell derived therefrom, with an agent that inhibits the synthesis of a pigment such as carotenoids, which can absorb light generated by the luciferase.","For example, the genetically modified plant cell can be contacted with an agent such as norfluorazon, which reduces or inhibits chlorophyll production in the plants, thus allowing increased emission of photons from the plant cell, or from a trrnsgenic plant derived from the plant cell.","A visibly bioluminescent genetically modified plant cell made according to a method of the invention generally contains the transgene stably maintained in the plant cell genome.","Preferably, the transgene is integrated into the plant cell genome.","As such, the present invention also relates to a genetically modified plant cell produced by a method of the invention, as well as to a transgenic plant containing or generated from such a genetically modified plant cell.","The present invention also relates to kit, which contains a genetically modified plant cell of the invention, or a derivative of the genetically modified plant cell, for example, a transgenic plant derived from the genetically modified plant cell, or a cell, a tissue, or an organ of such a transgenic plant.","As such, a kit of the invention can contain one or more flowers, bracts, leaves or other tissues or organs, which, upon contact with luciferin, are visibly luminescent.","Accordingly, a kit of the invention can further include an amount of luciferin sufficient for generating visible bioluminescence of the genetically modified plant cell or the derivative of the genetically modified plant cell, and also can include a plurality of such amounts of luciferin, thus allowing the generation of visible bioluminesce a number of times, as desired.","In addition, a kit of the invention can include an amount of a inhibitor that reduces or inhibits pigment synthesis in the genetically modified plant cell or the derivative of the genetically modified plant cell.","For example, the kit can contain a carotenoid synthesis inhibitor such as norfluorazon, which reduces or inhibits chlorophyll production.","In another embodiment, a kit of the invention contains one or more cuttings, seeds, or other portion or derivative of a visibly bioluminescent plant of the invention such that a visibly luminescent transgenic plant can be grown therefrom.","As such, the kit also can include reagents for growing a transgenic plant from the cutting or seed, including, for example, a suitable plant food or other nutrient source required for growth of the particular plant.","In addition, the kit can include an amount of a carotenoid inhibitor sufficient for reducing or inhibiting chlorophyll production in a trrnsgenic plant grown from the cutting or seed; and/or can include an amount of luciferin sufficient for generating visible bioluminescence of a transgenic plant grown from the cutting or seed."],["BACKGROUND OF THE INVENTION FIELD OF THE INVENTION The present invention relates generally to plant biology and more specifically to plants that are genetically modified to exhibit bioluminescence, and to compositions and methods for making such genetically modified bioluminescent plants.","BACKGROUND INFORMATION Plants and plant products provide the primary sustenance, either directly or indirectly, for all animal life, including humans.","For the majority of the world's human population and for many animals, plants and plant products provide the sole source of nutrition.","As the world population increases, the best hope to prevent widespread famine is to increase the quantity and improve the quality of food crops.","Throughout history, a continual effort has been made to increase the yield and nutritious value of food crops.","For centuries, plants having desirable characteristics such as greater resistance-to drought conditions or increased size of fruit were crossbred and progeny plants exhibiting the desired characteristics were selected and used to produce seed or cuttings for propagation.","Using such classical genetic methods, plants having, for example, greater disease resistance, increased yield, and better flavor have been obtained.","More recently, the methods of genetic engineering have been applied to plant biology, thus expediting the generation of plants having desired characteristics.","For example, genetically engineered tomato plants have been generated such that the tomatoes do not continue to ripen after they have been picked from the plant.","As a result, there is no need to pick unripe tomatoes and hope that they attain an optimum level of ripeness at the time the consumer is ready to eat them.","Instead, the tomatoes can be picked at a time when they are near their optimal ripeness, then shipped to consumers for consumption.","Genetically engineered plants that are less susceptible to freezing or to damage by biological pests also have been generated.","The availability of such plants has been a boon to the agricultural industry, and provides a measure of certainty that sufficient quantities of food will remain available in the future.","Methods of genetic engineering also have impacted the ornamental plant industry.","In addition to protecting ornamental plants from environmental stresses or biological pests, genetic engineering has been used to produce ornamental plants having new and interesting characteristics.","For example, genetic engineering has been used to generate roses that are blue in color.","The ability to generate ornamental plants having new and unusual characteristics provides a means to expand the ornamental plant market and, therefore, the economy in general.","Thus, a need exists to identify interesting and unusual traits that can be expressed by a plant and can create a new market in the ornamental plant industry.","The present invention satisfies this need and provides additional advantages.","SUMMARY OF THE INVENTION The present invention relates to a genetically modified plant cell that exhibits bioluminescence that is visible to the naked eye.","Such a genetically modified plant cell contains a heterologous nucleotide sequence that encodes a bioluminescent polypeptide, which, upon expression, can result in the production of at least about 750,000 photons of visible light/mm2/second by the plant cell.","The bioluminescent polypeptide can be any polypeptide that is bioluminescent, or can act in concert with a second molecule such that visible light is produced.","In one embodiment, the genetically modified plant cell contains a heterologous nucleotide sequence encoding luciferase or a luciferase variant, for example, luc+.","Upon contact of a genetically modified plant expressing luciferase or a variant thereof with luciferin, a sufficient amount of visible light is produced such that the light is visible to the naked eye.","Such a genetically modified plant cell generally expresses at least about 100 pg luc+/μg protein as determined by western blot analysis such that, upon contact of the plant cell with luciferin, the cell is visibly bioluminescent.","In another embodiment, the genetically modified plant cell, or the plant cell from which the genetically modified plant cell is derived, is contacted with an agent that reduces or inhibits pigment formation in the plant, for example, carotenoid synthesis, thereby reducing or inhibiting chlorophyll synthesis.","Since chlorophyll in a plant cell can absorb photons generated by the luciferase in the genetically modified plant cell, contact of the genetically modified plant cell, or a transgenic plant derived therefrom, with an amount of a carotenoid inhibitor sufficient to reduce or inhibit chlorophyll synthesis provides a means to enhance photon emission from the plant cell and, therefore, enhance the visible bioluminescence.","In still another embodiment, a visibly bioluminescent transgenic plant is generated from the genetically modified plant cell containing a heterologous nucleotide sequence encoding a bioluminescent polypeptide.","As such, the invention relates to such a transgenic plant, as well as to a plant cell or tissue obtained from the transgenic plant, a seed produced by the transgenic plant, and a cDNA or genomic DNA library prepared from the transgenic plant or from a plant cell or plant tissue obtained from the transgenic plant.","The transgenic plant can be a monocot, or can be a dicot, for example, an angiosperm such as a cereal plant, a leguminous plant, an oilseed plant, a hardwood tree, or an ornamental plant such as a petunia, an orchid, a carnation or the like.","The present invention also relates to a recombinant nucleic acid molecule, which comprises a translational enhancer, which enhances translation in a plant cell, operatively linked to a nucleotide sequence encoding a bioluminescent polypeptide.","The translational enhancer can be any translational enhancer, for example, a plant potyvirus translational enhancer such as tobacco etch virus (TEV) translational enhancer, a tobacco mosaic virus omega translational enhancer, or a translational enhancer comprising a Kozak sequence.","The encoded bioluminescent polypeptide can be a luciferase polypeptide or variant thereof, for example, the luc+ luciferase variant.","A recombinant nucleic acid molecule of the invention also can contain a transcriptional regulatory element, i.e., promoter, enhancer, etc., that enhances transcription in a plant cell, for example, a plant virus enhancer such as a cauliflower mosaic virus (CaMV) 35S enhancer, or a regulatory element from any other organism, for example, an actin 2 regulatory element or a metallothionein regulatory element, which comprises a promoter and, optionally, an enhancer, wherein the transcriptional regulatory element is operatively linked to the plant translational enhancer and nucleotide sequence encoding the bioluminescent polypeptide.","In one embodiment, a recombinant nucleic acid molecule contains, in operative linkage, a CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and a CaMV 35S terminator.","In another embodiment, the CaMV 35S enhancer is a dual CaMV 35S enhancer, and the luciferase polypeptide or variant thereof is the luc+ luciferase variant.","Such a recombinant nucleic acid molecule is exemplified herein by the nucleotide sequence set forth as nucleotides 8366 to 11,113 of SEQ ID NO:2.In still other embodiments, the transcriptional regulatory element in a recombinant nucleic molecule of the invention is a metallothionein regulatory element or an actin 2 regulatory element, and the translational enhancer is an omega enhancer, for example, in a recombinant nucleic acid molecule having a sequence set forth as nucleotides 8340 to 12,098 of SEQ ID NO:4 or as nucleotides 6 to 3570 of SEQ ID NO:5.The present invention further relates to a vector containing a recombinant nucleic acid molecule comprising a translational enhancer active in a plant operatively linked to a nucleotide sequence encoding a bioluminescent polypeptide.","Preferably, the vector is an expression vector.","In one embodiment, the recombinant nucleic acid molecule contained in the vector comprises, in operative linkage, a CAMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and a CaMV 35S terminator.","In another embodiment, the recombinant nucleic acid molecule contained in the vector comprises, in operative linkage, a dual CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luc+ luciferase variant, and a CaMV 35S terminator.","In still another embodiment, the recombinant nucleic acid molecule contained in the vector comprises the nucleotide sequence set forth as nucleotides 8366 to 11,113 of SEQ ID NO:2.In yet another embodiment, the transcriptional regulatory element in the recombinant nucleic acid molecule comprises an actin 2 promoter or a metallothionein promoter, and the translation enhancer is an omega enhancer, for example, in the recombinant nucleic acid molecules set forth as nucleotides 8340 to 12,098 of SEQ D) NO:4 or as nucleotides 6 to 3570 of SEQ ID NO:5.Vectors of the invention are exemplified herein by those as set forth in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:4 and SEQ ID NO:5.The present invention also provides a cell containing a recombinant nucleic acid molecule of the invention, or containing a vector of the invention.","The present invention further relates to a method of producing a genetically modified plant cell that is visibly bioluminescent.","A method of the invention can be performed, for example, by introducing a transgene comprising a nucleotide sequence encoding a bioluminescent polypeptide into a plant cell, whereby the bioluminescent polypeptide is expressed at a level that produces at least about 750,000 photons of visible light/mm2/second.","The transgene useful in a method of making a genetically modified plant cell that is visibly bioluminescent generally includes a translational enhancer that is active in a plant, for example, a plant translational enhancer such as a plant potyvirus translational enhancer (e.g., a TEV translational enhancer), a tobacco mosaic virus (TMV) translational enhancer such as the omega enhancer, operatively linked to the nucleotide sequence encoding a bioluminescent polypeptide, for example, a luciferase polypeptide or variant thereof.","The transgene also can contain a plant virus enhancer, for example, a CaMV 35S enhancer, which is operatively linked to the plant translational enhancer and nucleotide sequence encoding a bioluminescent polypeptide.","In one embodiment, a transgene useful in making a visibly luminescent genetically modified plant cell contains, in operative linkage, a dual CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and a CaMV 35S terminator.","In another embodiment, the nucleotide sequence encodes a luc+ luciferase variant polypeptide.","Such transgenes are exemplified by the nucleotide sequences set forth as nucleotides 8366 to 11,113 of SEQ ID NO:2, nucleotides 8340 to 12,098 of SEQ ID NO:4, and nucleotides 6 to 3570 of SEQ ID NO:5.A method of the invention can further include contacting the plant cell, or the genetically modified plant cell derived therefrom, with an agent that inhibits the synthesis of a pigment such as carotenoids, which can absorb light generated by the luciferase.","For example, the genetically modified plant cell can be contacted with an agent such as norfluorazon, which reduces or inhibits chlorophyll production in the plants, thus allowing increased emission of photons from the plant cell, or from a trrnsgenic plant derived from the plant cell.","A visibly bioluminescent genetically modified plant cell made according to a method of the invention generally contains the transgene stably maintained in the plant cell genome.","Preferably, the transgene is integrated into the plant cell genome.","As such, the present invention also relates to a genetically modified plant cell produced by a method of the invention, as well as to a transgenic plant containing or generated from such a genetically modified plant cell.","The present invention also relates to kit, which contains a genetically modified plant cell of the invention, or a derivative of the genetically modified plant cell, for example, a transgenic plant derived from the genetically modified plant cell, or a cell, a tissue, or an organ of such a transgenic plant.","As such, a kit of the invention can contain one or more flowers, bracts, leaves or other tissues or organs, which, upon contact with luciferin, are visibly luminescent.","Accordingly, a kit of the invention can further include an amount of luciferin sufficient for generating visible bioluminescence of the genetically modified plant cell or the derivative of the genetically modified plant cell, and also can include a plurality of such amounts of luciferin, thus allowing the generation of visible bioluminesce a number of times, as desired.","In addition, a kit of the invention can include an amount of a inhibitor that reduces or inhibits pigment synthesis in the genetically modified plant cell or the derivative of the genetically modified plant cell.","For example, the kit can contain a carotenoid synthesis inhibitor such as norfluorazon, which reduces or inhibits chlorophyll production.","In another embodiment, a kit of the invention contains one or more cuttings, seeds, or other portion or derivative of a visibly bioluminescent plant of the invention such that a visibly luminescent transgenic plant can be grown therefrom.","As such, the kit also can include reagents for growing a transgenic plant from the cutting or seed, including, for example, a suitable plant food or other nutrient source required for growth of the particular plant.","In addition, the kit can include an amount of a carotenoid inhibitor sufficient for reducing or inhibiting chlorophyll production in a trrnsgenic plant grown from the cutting or seed; and/or can include an amount of luciferin sufficient for generating visible bioluminescence of a transgenic plant grown from the cutting or seed.","BRIEF DESCRIPTION OF THE FIGURES FIG.","1 shows a map of the pTK1829 plasmid.","Restriction endonuclease sites are indicated, as are the first nucleotide positions of the sites, except that the first and last nucleotide positions of the multiple cloning site are indicated.","“luc+” indicates region encoding variant luciferase polypeptide; “Amp” indicates ampicillin-resistance gene (see, also, SEQ ID NO:1).","FIG.","2 shows a map of the pPZPTK1829 binary vector plasmid.","Restriction endonuclease sites and nucleotide positions are indicated as in FIG.","1.“luc+” indicates region encoding variant luciferase polypeptide; “Spec” and “Gent-R” indicate the genes conferring resistance to spectinomycin and gentamycin, respectively.","“Ori” indicates the plasmid origin of replication.","“LB” and “RB” indicate left border and right border, respectively, of the binary vector that allow transfer of the insert into the plant (see, also, SEQ ID NO:2).","FIG.","3 shows a map of the MT-OM-LUC+ binary vector plasmid.","Restriction endonuclease sites and nucleotide positions are indicated as in FIG.","1.“luc+”, “Spec”, “Gent-R”, “Ori”, and “LB” and “RB” are as in FIG.","2.“MT” indicates the metallothionein transcriptional regulatory element; “Ome” indicates the omega translational enhancer; and “E9 3′” indicates the RbcS E9 polyA region (see, also, SEQ ID NO:4).","FIG.","4 shows a map of the ACT-OM-LUC+ binary vector plasmid.","Restriction endonuclease sites and nucleotide positions are indicated as in FIG.","1.“luc+”, “Spec”, “Gent-R”, “ori”, “LB” and “RB” are as in FIG.","2.“Ome” and “E9 3′” are as in FIG.","3; “ACT” indicates the actin 2 transcriptional regulatory element (see, also, SEQ ID NO:5).","DETAILED DESCRIPTION OF THE INVENTION The present invention provides genetically modified plant cells that are visibly bioluminescent due to expression of heterologous polypeptide, and visibly bioluminescent transgenic plants comprising or generated from such genetically modified plant cells.","As disclosed herein, the transgenic plants of the invention exhibit bioluminescence that is visible to the human eye.","Prior to the present disclosure, plant cells could be genetically modified to express heterologous bioluminescent polypeptides such as luciferase, but light gathering or amplifying instrumentation was required to detect the bioluminescence; one could not simply look at plant comprising such cells directly and detect the bioluminescence.","It is noted that there is a report on the world wide web, at the URL “hybridorchids.com”, of a bioluminescent orchid.","However, no published information for making such orchids is available and the information on the website has not been updated since 2000.The present invention provides compositions and methods that obviate the previous limitations that prevented the generation of visibly detectable plants and plant cells.","As disclosed herein, the visibly bioluminescent genetically modified plant cells and transgenic plants of the invention are useful as research tools, and have great ornamental value.","A genetically modified plant cell or transgenic plant of the invention is visibly bioluminescent due to expression in the plant cell, or in plant cells of the transgenic plant, of a bioluminescent polypeptide.","As used herein, the term “bioluminescent polypeptide” refers to a polypeptide that can be expressed in a living organism and exhibits or effects the emission of visible light.","A bioluminescent polypeptide can inherently emit visible light such as fluorescence, or can be contacted with a physical or chemical agent such that a reaction occurs and visible light is produced.","For example, a genetically modified plant cell can contain a heterologous nucleotide sequence encoding luciferase such that, upon expression of the luciferase and contact of the plant cell with luciferin, a chemiluminescent reaction occurs and visible light is emitted.","As used herein, the term “visible light” refers to electromagnetic radiation associated with the visible portion of the electromagnetic spectrum, generally electromagnetic radiation having a wavelength between about 100 to 1000 nanometers (nm), and particularly between about 350 to 750 nm.","Luciferase polypeptides and luciferase variants are examples of bioluminescent polypeptides that are particularly useful for practicing the methods and producing the compositions of the invention.","As used herein, the term “luciferase variant” refers to a modified luciferase polypeptide that contains one or more amino acid additions, deletions or substitutions as compared to a corresponding wild type luciferase polypeptide, and that, when contacted with an appropriate substrate such as luciferin, undergoes a reaction that results in-the production of visible light.","Luciferases include bacterial luciferases such as Vibrio harveyi or V. fisheri luciferase, marine ostracod crustacean luciferases such a Vargula hilgendorii luciferase, dinoflagellate luciferases such as Gonyaulax ploydra luciferase, coelenterate luciferases such as Renilla renifornis (sea pansy) luciferase, or Coleoptera (beetle) luciferases such as an Elateridae (click beetle) family luciferase, a Phengodidae (glow worm) family luciferase, or a Lampyridae (firefly) luciferase, for example, a Photintus carolinus or P. pyralis luciferase, and the like, including modified forms thereof (see, for example, U.S. Pat.","No.","4,968,613; U.S. Pat.","No.","5,221,623; U.S. Pat.","No.","5,229,285; U.S. Pat.","No.","5,330,906; U.S. Pat.","No.","5,604,123; U.S. Pat.","No.","5,618,722; U.S. Pat.","No 5,674,713; U.S. Pat.","No.","5,814,465; and U.S. Pat.","No.","5,843,746, each of which is incorporated herein by reference).","A luciferase variant is exemplified by the firefly luciferase variant, luc+ (U.S. Pat.","No.","5,670,356; Sherf and Wood, Promega Notes 49:14, 1994, each of which is incorporated herein by reference), which lacks a peroxisome localizing domain, can be particularly useful for preparing a genetically modified plant cell or visibly bioluminescent transgenic plant of the invention.","Polynucleotide sequences encoding luciferases and mutant luciferases are well known (see above references) and vectors containing such polynucleotides are commercially available (Promega; Madison Wis.).","As used herein, the term “visibly bioluminescent” refers to the emission of a sufficient amount of visible light such that it can be seen by a human eye.","As such, the use of light gathering, light amplifying, light enhancing or light accumulating instrumentation is not required to detect the bioluminescence, although such instrumentation can be used, for example, to quantitate the amount of visible light emitted.","It should be recognized that a visibly bioluminescent plant cell may not be visible to the unaided eye due to its small size, but can be seen using a magnifying glass, microscope, or the like.","Such a plant cell is considered to be visibly bioluminescent for purposes of the present invention because light gathering, accumulating or amplifying instrumentation such as a luminometer, photomultiplier, image enhancing video system, spectrophotometer, film emulsion, digitizing equipment, or the like is not required.","The light produced by a visibly bioluminescent transgenic plant can be seen by a person or other mammal looking at the plant, particularly when the plant is in a dark environment, for example, in a darkened room or outside at night.","In addition, a bioluminescent plant of the invention can be photographed using a camera, including any generally used camera such as a 35 mm single lens reflex camera, a digital camera, a Polaroid® camera, and the like, such that a picture of the glowing plant can be obtained.","As disclosed herein, a genetically modified plant cell, or a plant, plant tissue or plant organ comprising the plant cell that emits at least about 750,00 photons of visible light/mm2second, generally at least about 850,000 photons of visible light/mm2/second, and particularly at least about 1×106 (one million) photons of visible light/mm2/second is visibly bioluminescent (see Example 2C, Table 4).","As disclosed herein, contact of a genetically modified plant cell of the invention, or of a plant cell from which the genetically modified plant cell is derived, or of a transgenic plant of the invention with an agent that reduces or inhibits the synthesis of one or more pigments in the plant or plant cell.","By reducing the amount of pigmentation of a plant, or a portion of the plant such as the leaves or flowers, absorption of bioluminescence by the pigment is reduced, thereby enhancing the emission of photons from the plant and, therefore, the visible bioluminescence.","For example, contact of the genetically modified plant cell or a transgenic plant of the invention with an inhibitor of carotenoid synthesis can reduce or inhibit chlorophyll synthesis in the plant, thus reducing absorption of photons by chlorophyll in the plant and allowing greater emission of the photons from the plant.","Inhibitors of carotenoid synthesis in plants are well known and include herbicides such as norfluorazone, which is commercially available as ZORIAL® herbicide (Syngenta Crop Production; Greensboro N.C.).","In another embodiment, a visibly bioluminescent transgenic plant is generated from the genetically modified plant cell containing a heterologous nucleotide sequence encoding a bioluminescent polypeptide.","As such, the invention relates to such a transgenic plant, as well as to a plant cell or tissue obtained from the transgenic plant, a seed produced by the transgenic plant, and a cDNA or genomic DNA library prepared from the trrnsgenic plant or from a plant cell or plant tissue obtained from the transgenic plant.","The transgenic plant can be a monocot, or can be a dicot, for example, an angiosperm such as a cereal plant, a leguminous plant, an oilseed plant, a hardwood tree, or an ornamental plant such as a petunia, a carnation or the like.","In one embodiment, plants useful for purposes of the present invention include orchids.","In another embodiment, plants useful for purposes of the present invention exclude orchids.","The present invention provides a recombinant nucleic acid molecule, which includes a heterologous nucleotide sequence encoding a bioluminescent polypeptide operatively linked to one or more transcriptional or translational regulatory elements, or a combination thereof.","As used herein, the term “heterologous” is used in a comparative sense with respect to a nucleotide sequence encoding a bioluminescent polypeptide to indicate either that the nucleotide sequence is not an endogenous nucleotide sequence in a plant cell into which it is to be introduced, or that the nucleotide sequence is part of a construct such that it is in a form other than it normally would be found in a plant cell in nature.","For example, a heterologous nucleotide sequence can encode a firefly luciferase, which is not normally expressed in a plant cell and, therefore, is heterologous with respect to the plant cell.","The regulatory element or combination of regulatory elements is selected such that the bioluminescent polypeptide can be expressed in a plant cell at a level sufficient for the plant cell, or a plant comprising the cell, to visibly bioluminesce.","The term “regulatory element” is used broadly herein to refer to a nucleotide sequence that, when operatively linked to a second, expressible nucleotide effects transcription or translation of the nucleotide sequence such that a ribonucleic acid (RNA) molecule or polypeptide, respectively, can be generated, or encodes a domain that confers a desirable characteristic on a polypeptide containing the domain.","For purposes of the present invention, a regulatory element generally increases the amount of transcription or translation of an operatively linked nucleotide sequence encoding the bioluminescent polypeptide, or provides a means to localize a bioluminescent polypeptide to a particular location in a cell, particularly a plant cell.","Transcriptional and translational regulatory elements are well known and include promoters, enhancers, 3′-untranslated or 5′-untranslated sequences of transcribed sequence, for example, a poly-A signal sequence or other transcription termination signal, or other protein or RNA stabilizing elements, or other gene expression control elements known to regulate gene expression or the amount of expression of a gene product.","A regulatory element useful for constructing a recombinant nucleic acid molecule of the invention can be isolated from a naturally occurring genomic DNA sequence or can be synthetic, for example, a synthetic promoter.","Transcriptional regulatory elements can be constitutively expressed regulatory elements, which maintain gene expression at a relatively constant level of activity, or can be inducible regulatory elements.","Constitutively expressed regulatory elements can be expressed in any cell type, for example an actin promoter such as an actin 2 promoter, or an elongation factor (EF) promoter such as an EF1α promoter; or can be a tissue specific regulatory element, which is expressed only in one or a few specific cell types, a phase specific regulatory element, which is expressed only during particular developmental or growth stages of a plant cell, or the like.","A regulatory element such as a tissue specific or phase specific regulatory element or an inducible regulatory element useful in constructing a recombinant nucleic acid molecule of the invention can be a regulatory element that generally, in nature, is found in a plant genome.","However, the regulatory element also can be from an organism other than a plant, including, for example, from a plant virus, an animal virus, or a cell from an animal or other multicellular organism.","As used herein, the term “tissue specific or phase specific regulatory element” means a nucleotide sequence that effects transcription in only one or a few cell types, or only during one or a few stages of the life cycle of a plant, for example, only for a period of time during a particular stage of growth, development or differentiation.","The terms “tissue specific” and “phase specific” are used together herein in referring to such a regulatory element because a single regulatory element can have characteristics of both types of regulatory elements.","For example, a regulatory element active only during a particular stage of plant development also can be expressed only in one or a few types of cells in the plant during the particular stage of development.","A tissue specific or phase specific regulatory element can be useful for producing a genetically modified plant that is bioluminescent only in particular organs or tissues, or only at a particular developmental stage.","Numerous tissue specific or phase specific regulatory elements have been described and are known and available to those in the art.","Such tissue or phase specific regulatory elements include, for example, the AGL8/FRUITFULL regulatory element, which is activated upon floral induction (Hempel et al., Development 124:3845-3853, 1997, which is incorporated herein by reference); root specific regulatory elements such as the regulatory elements from the RCP1 gene and the LRP1 gene (Tsugeki and Fedoroff, Proc.","Natl.","Acad.","Sci., USA 96:12941-12946, 1999; Smith and Fedoroff, Plant Cell 7:735-745, 1995, each of which is incorporated herein by reference); flower specific regulatory elements such as the regulatory elements from the LEAFY gene and the APETELA1 gene (Blazquez et al., Development 124:3835-3844, 1997, which is incorporated herein by reference; Hempel et al., supra, 1997); seed specific regulatory elements such as the regulatory element from the oleosin gene (Plant et al., Plant Mol.","Biol.","25:193-205, 1994, which is incorporated herein by reference), and dehiscence zone specific regulatory element.","Additional tissue specific or phase specific regulatory elements include the Zn13 promoter, which is a pollen specific promoter (Hamilton et al., Plant Mol.","Biol.","18:211-218, 1992, which is incorporated herein by reference); the UNUSUAL FLORAL ORGANS (UFO) promoter, which is active in apical shoot meristem; the promoter active in shoot meristems (Atanassova et al., Plant J.","2:291, 1992, which is incorporated herein by reference), the cdc2a promoter and cyc07 promoter (see, for example, Ito et al., Plant Mol.","Biol.","24:863, 1994; Martinez et al., Proc.","Natl.","Acad.","Sci., USA 89:7360, 1992; Medford et al., Plant Cell 3:359, 1991; Terada et al., Plant J.","3:241, 1993; Wissenbach et al., Plant J.","4:411, 1993, each of which is incorporated herein by reference); the promoter of the APETELA3 gene, which is active in floral meristems (Jack et al., Cell 76:703, 1994, which is incorporated herein by reference; Hempel et al., supra, 1997); a promoter of an agamous-like (AGL) family member, for example, AGL8, which is active in shoot meristem upon the transition to flowering (Hempel et al., supra, 1997); floral abscission zone promoters; L1-specific promoters; and the like.","Such tissue specific or phase specific regulatory elements can be used to construct a recombinant nucleic acid useful for producing, for example, a transgenic plant that is visibly bioluminescent in one or a few selected tissues or organs such as in flowers or in leaves, or that are expressed only during a particular stage of development such as during flowering.","Inducible regulatory elements also are useful for constructing a recombinant nucleic acid molecule of the invention.","As used herein, the term “inducible regulatory element” means a regulatory element that, when exposed to an inducing agent, effects an increased level of transcription of an operatively linked nucleotide sequence encoding the bioluminescent polypeptide as compared to the level of transcription, if any, in the absence of an inducing agent.","Inducible regulatory elements can be those that have no basal or constitutive activity and only effect transcription upon exposure to an inducing agent, or those that effect a basal or constitutive level of transcription, which is increased upon exposure to an inducing agent.","Inducible regulatory elements that effect a basal or constitutive level of expression generally are particularly useful where the induced level of transcription is substantially greater than the basal or constitutive level of expression, for example, at least about two-fold greater, or at least about five-fold greater.","Particularly useful inducible regulatory elements do not have a basal or constitutive activity, or increase the level of transcription at least about ten-fold greater than a basal or constitutive level of transcription associated with the regulatory element.","The term “inducing agent” is used to refer to a chemical, biological or physical agent that effects transcription from an inducible regulatory element.","In response to exposure to an inducing agent, transcription from the inducible regulatory element generally is initiated de novo or is increased above a basal or constitutive level of expression.","Such induction can be identified using the methods disclosed herein, including detecting an increased level of mRNA encoding the bioluminescent polypeptide.","The use of an inducible regulatory element in a recombinant nucleic acid molecule of the invention provides a means to express the bioluminescent polypeptide only at a desired time, thus preventing extraneous transcriptional or translational activity in the plant cell.","An inducing agent useful in a method of the invention is selected based on the particular inducible regulatory element.","For example, the inducible regulatory element can be a metallothionein (MT) regulatory element such as an MT2B regulatory element, a copper inducible regulatory element, or a tetracycline inducible regulatory element, the transcription from which can be effected in response to various metal ions, to copper or to tetracycline, respectively (Furst et al., Cell 55:705-717, 1988; Mett et al., Proc.","Natl.","Acad.","Sci., USA 90:4567-4571, 1993; Gatz et al., Plant J.","2:397-404, 1992; Roder et al., Mol.","Gen. Genet.","243:32-38, 1994, each of which is incorporated herein by reference).","The inducible regulatory element also can be an ecdysone regulatory element or a glucocorticoid regulatory element, the transcription from which can be effected in response to ecdysone or other steroid (Christopherson et al., Proc.","Natl.","Acad.","Sci., USA 89:6314-6318, 1992; Schena et al., Proc.","Natl.","Acad.","Sci., USA 88:10421-10425, 1991, each of which is incorporated herein by reference).","In addition, the regulatory element can be a cold responsive regulatory element or a heat shock regulatory element, the transcription of which can be effected in response to exposure to cold or heat, respectively (Takahashi et al., Plant Physiol.","99:383-390, 1992, which is incorporated herein by reference).","Additional regulatory elements useful in the methods or compositions of the invention include, for example, the spinach nitrite reductase gene regulatory element (Back et al., Plant Mol.","Biol.","17:9, 1991, which is incorporated herein by reference); a light inducible regulatory element (Feinbaum et al., Mol.","Gen. Genet.","226:449, 1991; Lam and Chua, Science 248:471, 1990, each of which is incorporated herein by reference), a plant hormone inducible regulatory element (Yamaguchi-Shinozaki et al., Plant Mol.","Biol.","15:905 (1990); Kares et al., Plant Mol.","Biol.","15:225 (1990), each of which is incorporated herein by reference), and the like.","A circadian rhythm regulatory element such as a cab2 promoter (see, for example, Millar et al., Plant Cell 4:1075-1087, 1992, which is incorporated herein by reference) can also be considered an example of an inducible regulatory element, since expression from such a promoter appears to occur in response to a factor associated with a time of day.","A regulatory element operatively linked to a nucleotide sequence encoding a bioluminescent polypeptide also can comprise an enhancer, including a transcriptional enhancer, translational enhancer, or both.","A transcriptional enhancer useful for purposes of the present invention can be any enhancer generally associated with a plant gene or an enhancer from another organism that has enhancer activity in a plant cell, for example, a plant virus enhancer, or an enhancer from a gene expressed in an invertebrate cell, or a vertebrate cell such as a mammalian cell, or an enhancer of a virus that infects such cells.","A cauliflower mosaic virus (CaMV) 35S enhancer is an example of a plant virus enhancer that is useful for increasing the transcriptional activity of a nucleotide sequence to which it is operatively linked.","Thus, in one embodiment, a recombinant nucleic acid molecule of the invention-contains a CaMV 35S enhancer operatively linked to a nucleotide sequence encoding the bioluminescent polypeptide.","Preferably, the CaMV 35S enhancer is a dual enhancer, comprising two copies of the CaMV 35S enhancer nucleotide sequence.","An actin 2 regulatory element provides another example of a transcriptional regulatory element, comprising a promoter and an enhancer, that is useful in constructing a composition of the invention or practicing a method of the invention (see nucleotides 6 to 1145 of SEQ ID NO:5).","A metallothionein regulatory element provides another example of a transcriptional regulatory element useful for purposes of the present invention, and provides the additional advantage that it includes an inducible enhancer (see nucleotides 11,000 to 12,098 of SEQ ID NO:4).","A translational enhancer, which enhances translation of a polypeptide in a plant cell (also referred to generally herein as a plant translational enhancer), useful in a recombinant nucleic acid molecule of the invention similarly can be any translational enhancer that generally is associated with a plant gene or with a gene of another organism.","Examples of a plant translational enhancer include a plant potyvirus translational enhancer such as the tobacco etch virus (TEV) translational enhancer, and a tobacco mosaic virus translational enhancer such as the omega enhancer (see, for example, nucleotides 1169 to 1235 of SEQ ID NO:5; see, also, Carrington and Freed, J. Virol.","64:1590, 1990; Gallie et al., Plant Cell 1:301-311, 1989, each of which is incorporated herein by reference), as well as the nucleotide sequence beginning about 101 nucleotide upstream of the potato virus S (PSV) coat protein gene (Turner and Foster, Arch.","Virol.","142:167-175, 1997, which is incorporated herein by reference).","A recombinant nucleic acid molecule of the invention can also contain regulatory elements required for termination of transcription, for example, a polyadenylation signal.","A CaMV 35S terminator is an example of a transcription termination signal useful in constructing a recombinant nucleic acid molecule.","A recombinant nucleic acid molecule of the invention also can contain regulatory elements required for initiation or termination of translation or both, including, for example, an internal ribosome entry site (IRES), a Kozak sequence, a nucleotide sequence encoding an initiator methionine residue, and a nucleotide sequence encoding a STOP codon, which terminates translation.","The regulatory element also can be a nucleotide sequence encoding a leader sequence, signal peptide, cell compartmentalization domain, or the like, and can be included in the construct, for example, where it is desired to localize the bioluminescent polypeptide to a particular compartment such as the cytosol, nucleus, a chloroplast, or another subcellular organelle in a plant cell.","Such intracellular localization can be particularly useful where the genetically modified cell, or plant containing the cell, is to be used as a research tool, for example, a particular intracellular compartment of a plant cell.","A translation initiation or termination element or other regulatory element can be contained in the nucleotide sequence encoding the bioluminescent polypeptide can be a regulatory element that is normally associated with the nucleotide sequence in nature and is a part of the nucleotide sequence as a consequence of its isolation from a natural source, or can be heterologous to the nucleotide sequence and operatively linked thereto.","Additional regulatory elements such as an intron sequence, for example, from Adh1 or bronzel, or a viral leader sequence, for example, from TMV, MCMV or AIVIV, also enhance expression of a nucleotide sequence encoding a bioluminescent polypeptide and, therefore, can be a component of a recombinant nucleic acid molecule of the invention.","As disclosed herein, combinations of regulatory elements can be selected such that a recombinant nucleic acid molecule comprising a nucleotide sequence encoding a bioluminescent polypeptide can be constructed, and wherein the recombinant nucleic acid molecule allows for expression of the bioluminescent polypeptide in a plant cell such that the plant cell is visibly bioluminescent.","A recombinant nucleic acid molecule of the invention is exemplified by, in operative linkage, a CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and a CaMV 35S terminator.","In one embodiment, the CaMV 35S enhancer comprises a dual CaMV 35S enhancer, and the luciferase polypeptide or variant thereof is a luc+luciferase variant.","In another embodiment, a recombinant nucleic acid molecule of the invention is exemplified by the nucleotide sequence set forth as about nucleotides 8366 to 11,113 of SEQ ID NO:2.A recombinant nucleic acid molecule of the invention also is exemplified by the sequence set forth as nucleotides 8340 to 12,098 of SEQ ID NO:4, which includes, in operative linkage, a metallothionein gene (MT2B) regulatory element comprising an enhancer and promoter (nucleotides 11,000 to 12,098 of SEQ ID NO:4), an omega translational enhancer (nucleotides 10,675 to 10,741 of SEQ ID NO:4), a nucleotide sequence encoding a luciferase polyp eptide or variant thereof such as luc+ (nucleotides 9015 to 10,667 or SEQ ID NO:4) and an RbcS E9 polyA region (nucleotides 8340 to 8981 of SEQ ID NO:4).","In addition, a recombinant nucleic acid molecule of the invention also is exemplified by the sequence set forth as nucleotides 6 to 3570 of SEQ ID NO:5, which includes, in operative linkage, an actin 2 regulatory element comprising an enhancer and promoter (nucleotides 6 to 1145 of SEQ ID NO:5), an omega translational enhancer (nucleotides 1169 to 1235 of SEQ ID NO:5), a nucleotide sequence encoding a luciferase polypeptide or variant thereof such as luc+ (nucleotides 1243 to 2895 of SEQ ID NO:5), and an RbcS E9 polyA region (nucleotides 2929 to 3570 of SEQ ID NO:5).","A recombinant nucleic acid molecule of the invention comprises a nucleotide sequence encoding a bioluminescent polypeptide operatively linked to one or more regulatory elements, for example, a translational enhancer.","As used herein, the term “operatively linked,” when used in reference to a regulatory element and a second nucleotide sequence, which can be another regulatory element or a coding or other sequence, means that the regulatory element is positioned with respect to the second nucleotide sequence such that the regulatory element effects its function with respect to the second nucleotide sequence in substantially the same manner as it does when the regulatory element is present in its natural position in a genome.","Transcriptional promoters, for example, generally act in a position and orientation dependent manner, and usually are positioned at or within about five nucleotides to about fifty nucleotides 5′ (upstream) of the start site of transcription of a gene in nature.","In comparison, enhancers can act in a relatively position or orientation independent manner, and can be positioned several hundred or thousand nucleotides upstream or downstream from a transcription start site, or in an intron within the coding region of a gene, yet still be operatively linked to the coding region so as to enhance transcription.","The relative positions and orientations of various regulatory elements, including the positioning of a transcribed regulatory sequence such as an IRES or a translated regulatory element such as a cell compartmentalization domain in an appropriate reading frame, are well known and methods for operatively linking such elements are routine in the art (see, for example, Sambrook et al., “Molecular Cloning: A laboratory manual” (Cold Spring Harbor Laboratory Press 1989); Ausubel et al., “Current Protocols in Molecular Biology” (John Wiley and Sons, Baltimore Md.","1987, and supplements through 1995), each of which is incorporated herein by reference).","As used herein, the term “nucleic acid molecule” or “polynucleotide” or “nucleotide sequence” means a sequence of two or more deoxyribonucleotides or ribonucleotides that are linked together by a phosphodiester bond.","As such, the terms include RNA and DNA, which can be a gene or a portion thereof, a cDNA, a synthetic polydeoxyribonucleic acid sequence, or the like, and can be single stranded or double stranded, as well as a DNA/RNA hybrid.","Furthermore, the terms are used herein to include naturally occurring nucleic acid molecules, which can be isolated from a cell, as well as synthetic molecules, which can be prepared, for example, by methods of chemical synthesis or by enzymatic methods such as by the polymerase chain reaction (PCR).","The term “recombinant” is used herein to refer to a nucleic acid molecule that is produced by linking together at least two nucleotide sequences that are not generally linked together in nature.","As such, a recombinant nucleic acid molecule encompassed within the present invention is distinguishable from a nucleic acid molecule that may be produced, for example, during meiosis.","In general, the nucleotides comprising a recombinant nucleic acid molecule, polynucleotide or nucleotide sequence are naturally occurring deoxyribonucleotides, such as adenine, cytosine, guanine or thymine linked to 2′-deoxyribose, or ribonucleotides such as adenine, cytosine, guanine or uracil linked to ribose.","However, a nucleic acid molecule or nucleotide sequence also can contain nucleotide analogs, including non-naturally occurring synthetic nucleotides or modified naturally occurring nucleotides.","Such nucleotide analogs are well known in the art and commercially available, as are polynucleotides containing such nucleotide analogs (Lin et al., Nucl.","Acids Res.","22:5220-5234, 1994; Jellinek et al., Biochemistry 34:11363-11372, 1995; Pagratis et al., Nature Biotechnol.","15:68-73, 1997, each of which is incorporated herein by reference).","Similarly, the covalent bond linking the nucleotides of a nucleotide sequence generally is a phosphodiester bond.","However, the covalent bond also can be any of numerous other bonds, including a thiodiester bond, a phosphorothioate bond, a peptide-like bond or any other bond known to those in the art as useful for linking nucleotides to produce synthetic polynucleotides (see, for example, Tam et al., Nucl.","Acids Res.","22:977-986, 1994; Ecker and Crooke, BioTechnology 13:351360, 1995, each of which is incorporated herein by reference).","The incorporation of non-naturally occurring nucleotide analogs or bonds linking the nucleotides or analogs can be particularly useful where the nucleic acid molecule is to be exposed to an environment that can contain a nucleolytic activity, including, for example, a plant tissue culture medium or in a plant cell since the modified molecules can be less susceptible to degradation.","A nucleotide sequence containing naturally occurring nucleotides and phosphodiester bonds, can be chemically synthesized or can be produced using recombinant DNA methods, using an appropriate polynucleotide as a template.","In comparison, a nucleotide sequence containing nucleotide analogs or covalent bonds other than phosphodiester bonds generally are chemically synthesized, although an enzyme such as T7 polymerase can incorporate certain types of nucleotide analogs into a polynucleotide and, therefore, can be used to produce such a polynucleotide recombinantly from an appropriate template (Jellinek et al., supra, 1995).","A recombinant nucleic acid molecule of the invention can be introduced into a cell as a naked DNA molecule, can be incorporated in a matrix such as a liposome or a particle such as a viral particle, or can be incorporated into a vector.","Incorporation of the polynucleotide into a vector can facilitate manipulation of the polynucleotide, or introduction of the polynucleotide into a plant cell.","Accordingly, the present invention also provides a vector containing a recombinant nucleic acid comprising a nucleotide sequence encoding a bioluminescent polypeptide operatively linked to one or more regulatory elements.","A vector can be derived from a plasmid or a viral vector such as a T-DNA vector (Horsch et al., Science 227:1229-1231 (1985), which is incorporated herein by reference).","If desired, the vector can comprise components of a plant transposable element, for example, a Ds transposon (Bancroft and Dean, Genetics 134:1221-1229, 1993, which is incorporated herein by reference) or an Spm transposon (Aarts et al., Mol.","Gen. Genet.","247:555-564, 1995, which is incorporated herein by reference).","In addition to containing a recombinant nucleic acid molecule of the invention, the vector also can contain various nucleotide sequences that facilitate, for example, rescue of the vector from a transformed plant cell; passage of the vector in a host cell, which can be a plant, animal, bacterial, or insect host cell; or expression of an encoding nucleotide sequence in the vector, including all or a portion of a rescued coding region.","As such, a vector can contain any of a number of additional transcription and translation elements, including constitutive and inducible promoters, enhancers, and the like (see, for example, Bitter et al., Meth.","Enzymol.","153:516-544, 1987).","For example, a vector can contain elements useful for passage, growth or expression in a bacterial system, including a bacterial origin of replication; a promoter, which can be an inducible promoter; and the like.","A vector also can contain one or more restriction endonuclease recognition and cleavage sites, including, for example, a polylinker sequence, to facilitate insertion or removal of a recombinant nucleic acid molecule.","In addition to a nucleotide sequence encoding a bioluminescent polypeptide, a recombinant nucleic acid molecule of the invention, or a vector containing such a recombinant nucleic acid molecule, can contain one or more other expressible nucleotide sequences encoding an RNA or a polypeptide of interest.","For example, the additional nucleotide sequence can encode an antisense nucleic acid molecule; an enzyme such as β-galactosidase, β-glucuronidase, luciferase, alkane phosphatase, glutathione S-transferase, chloramphenicol acetyltransferase, guanine xanthine phosphoribosyltransferase, and neomycin phosphotransferase; a viral polypeptide or a peptide portion thereof; or a growth factor or a hormone, which can be a plant growth factor or hormone.","Expression of such a nucleotide sequence can provide a means for selecting for a cell containing the construct, for example, by conferring a desirable phenotype to a plant cell containing the nucleotide sequence.","For example, the additional nucleotide sequence can be, or encode, a selectable marker, which, when present or expressed in a plant cell, provides a means to identify the plant cell containing the marker.","A selectable marker provides a means for screening a population of plants, or plant cells, to identify those having the marker and, therefore, a recombinant nucleic acid molecule comprising a nucleotide sequence encoding a bioluminescent polypeptide.","A selectable marker generally confers a selective advantage to the plant cell, or a plant containing the cell, for example, the ability to grow in the presence of a negative selective agent such as an antibiotic or herbicide.","A selective advantage also can be due, for example, to an enhanced or novel capacity to utilize an added compound as a nutrient, growth factor or energy source.","A selective advantage can be conferred by a single polynucleotide, or its expression product, or by a combination of polynucleotides whose expression in a plant cell gives the cell a positive selective advantage, a negative selective advantage, or both.","It should be recognized that expression of a bioluminescent polypeptide also provides a means to a select plant cell containing the encoding nucleotide sequence.","However, the use of an additional selectable marker, which, for example, allows a plant cell to survive under otherwise toxic conditions, provides a means to enrich for transformed plant cells containing the recombinant nucleic acid molecule.","Examples of selectable markers include those described above, as well as those that confer antimetabolite resistance, for example, dihydrofolate reductase, which confers resistance to methotrexate (Reiss, Plant Physiol.","(Life Sci.","Adv.)","13:143-149, 1994); neomycin phosphotransferase, which confers resistance to the aminoglycosides neomycin, kanamycin and paromycin (Herrera-Estrella, EMBO J.","2:987-995, 1983) and hygro, which confers resistance to hygromycin (Marsh, Gene 32:481-485, 1984), trpB, which allows cells to utilize indole in place of tryptophan; hisD, which allows cells to utilize histinol in place of histidine (Hartman, Proc.","Natl.","Acad.","Sci., USA 85:8047, 1988); mannose-6-phosphate isomerase which allows cells to utilize mannose (WO 94/20627); ornithine decarboxylase, which confers resistance to the ornithine decarboxylase inhibitor, 2-(difluoromethyl)-DL-ornithine (DFMO; McConlogue, 1987, In: Current Communications in Molecular Biology, Cold Spring Harbor Laboratory ed.","); and deaminase from Aspergillus terreus, which confers resistance to Blasticidin S (Tamura, Biosci.","Biotechnol.","Biochem.","59:2336-2338, 1995).","Additional selectable markers include those that confer herbicide resistance, for example, phosphinothricin acetyltransferase gene, which confers resistance to phosphinothricin (White et al., Nucl.","Acids Res.","18:1062, 1990; Spencer et al., Theor.","Appl.","Genet.","79:625-631, 1990), a mutant EPSPV-synthase, which confers glyphosate resistance (Hinchee et al., BioTechnology 91:915-922, 1998), a mutant acetolactate synthase, which confers imidazolione or sulfonylurea resistance (Lee et al., EMBO J.","7:1241-1248, 1988), a mutant psbA, which confers resistance to atrazine (Smeda et al., Plant Physiol.","103:911-917, 1993), or a mutant protoporphyrinogen oxidase (see U.S. Pat.","No.","5,767,373), or other markers conferring resistance to an herbicide such as glufosinate.","In addition, markers that facilitate identification of a plant cell containing the polynucleotide encoding the marker include, for example, luciferase (Giacomin, Plant Sci.","116:59-72, 1996; Scikantha, J. Bacteriol.","178:121, 1996), green fluorescent protein (Gerdes, FEBS Lett.","389:4447, 1996) or fl-glucuronidase (Jefferson, EMBO J.","6:3901-3907, 1997), and numerous others as disclosed herein or otherwise known in the art.","Such markers also can be used as reporter molecules.","As disclosed herein, a vector of the invention contains a recombinant nucleic acid molecule comprising one or more regulatory elements in operative linkage with a nucleotide sequence encoding a bioluminescent polypeptide.","For example, the recombinant nucleic acid molecule in a vector can contain, in operative linkage, a plant enhancer, plant promoter, a translational enhancer, a nucleotide sequence encoding a bioluminescent polypeptide, a translation termination sequence, and transcription termnination sequence.","It should be recognized that one or more of the regulatory elements initially can be a component of the vector or of the recombinant nucleic acid molecule.","In one embodiment, a vector of the invention contains a recombinant nucleic acid molecule comprising, in operative linkage, a CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and a CaMV 35S terminator.","In another embodiment, the recombinant nucleic acid molecule contained in the vector comprises, in operative linkage, a dual CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luc+ luciferase variant, and a CaMV 35S terminator.","Vectors of the invention are exemplified herein by a vector having the nucleotide sequence of SEQ ID NO:1 (see, also, FIG.","1), and by a vector having the nucleotide sequence of SEQ ID NO:2 (see, also, FIG.","2).","Additional vectors of the invention are exemplified by those set forth as SEQ ID NO:4 and SEQ ID NO:5.The present invention also provides a host cell containing a recombinant nucleic acid molecule of the invention, or containing a vector of the invention.","The present invention further provides a method of producing a genetically modified plant cell that is visibly bioluminescent.","As used herein, the term “genetically modified,” when used in reference to a plant cell, means that the plant cell contains an exogenously introduced nucleic acid molecule.","For purposes of the present invention, the introduced nucleotide sequence generally is a recombinant nucleic acid molecule comprising one or more regulatory elements operatively linked to a nucleotide sequence encoding a bioluminescent polypeptide, or a vector containing such a recombinant nucleic acid molecule.","The nucleic acid molecule can be transiently associated with the plant cell, but generally is stably associated with the genetically modified cell, as well as its progeny, either as an autonomously replicating molecule or due to integration into the plant cell genomic DNA or plastid DNA that is stably maintained in the plant cell.","If desired, a transgene comprising a recombinant nucleic acid molecule of the invention can be introduced into a plant genome in a site-specific matter, for example, by homologous recombination.","A method of the invention can be performed, for example, by introducing a transgene comprising a recombinant nucleic acid molecule of the invention into a plant cell such that the encoded bioluminescent polypeptide is expressed in the plant cell at a level that can produce at least about 750,000 to 1×106 photons of visible light/mm2/second.","As such, the present invention also provides a genetically modified plant cell produced by a method of the invention, as well as to a transgenic plant containing or generated from such a genetically modified plant cell, and to plant tissues, plant cells, plant organs and seeds produced by such a transgenic plant; as well as to a cDNA or genomic DNA library prepared from the transgenic plant, or from a plant cell or plant tissue obtained from the transgenic plant.","A method of the invention can further include contacting the plant cell, or the genetically modified plant cell derived therefrom, with an agent that reduces or inhibits pigment synthesis in the plant cell or in a tissue or organ of a transgenic plant of the invention, for example, the synthesis of an anthocyanin, which is a water-soluble red to blue plant pigment; a carotene, which is an orange-yellow pigment located in the chloroplasts; chlorophyll, which is the green pigment in green plants; xanthophyll, which is a yellow to colorless photosynthetic plant pigment, or any other pigment that can absorbs visible light, for example, lycopene, which is a red pigment found in tomatoes.","The agent that is selected for reducing or inhibiting pigmentation of a plant is based on the particular pigment, and that the pathway involved in synthesis of the pigment.","For example, an agent such as norfluorazon, which inhibits carotenoid synthesis, can be used to reduce the amount of chlorophyll in the plant cell, thus enhancing the emission of a greater number of photons from the plant cell, or from a transgenic plant derived from the plant cell.","The term “plant” is used broadly herein to include any plant at any stage of development, or to part of a plant, including a plant cutting, a plant cell, a plant cell culture, a plant organ, a plant seed, and a plantlet.","A plant cell is the structural and physiological unit of the plant, comprising a protoplast and a cell wall.","A plant cell can be in the form of an isolated single cell or aggregate of cells such as a friable callus, or a cultured cell, or can be part of higher organized unit, for example, a plant tissue, plant organ, or plant.","Thus, a plant cell can be a protoplast, a gamete producing cell, or a cell or collection of cells that can regenerate into a whole plant.","As such, a seed, which comprises multiple plant cells and is capable of regenerating into a whole plant, is considered plant cell for purposes of this disclosure.","A plant tissue or plant organ can be a seed, protoplast, callus, or any other groups of plant cells that is organized into a structural or functional unit.","Particularly useful parts of a plant include harvestable parts and parts usefull for propagation of progeny plants.","A harvestable part of a plant can be any useful part of a plant, for example, flowers, pollen, seedlings, tubers, leaves, stems, fruit, seeds, roots, and the like.","A part of a plant useful for propagation includes, for example, seeds, fruits, cuttings, seedlings, tubers, rootstocks, and the like.","A transgenic plant can be regenerated from a genetically modified plant cell, i.e., a whole plant can be regenerated from a plant cell; a group of plant cells; a protoplast; a seed; or a piece of a plant such as a leaf, a cotyledon or a cutting.","Regeneration from protoplasts varies from species to species of plants.","For example, a suspension of protoplasts can be made and, in certain species, embryo formation can be induced from the protoplast suspension, to the stage of ripening and germination.","The culture media generally contains various components necessary for growth and regeneration, including, for example, hormones such as auxins and cytokinins; and amino acids such as glutamic acid and proline, depending on the particular plant species.","Efficient regeneration will depend, in part, on the medium, the genotype, and the history of the culture.","If these variables are controlled, however, regeneration is reproducible.","Regeneration can occur from plant callus, explants, organs or plant parts.","Transformation can be performed in the context of organ or plant part regeneration.","(see Meth.","Enzymol.","Vol.","118; Klee et al.","Ann.","Rev.","Plant Physiol.","38:467 (1987), which is incorporated herein by reference).","Utilizing the leaf disk-transformation-regeneration method, for example, disks are cultured on selective media, followed by shoot formation in about two to four weeks (see Horsch et al., supra, 1985).","Shoots that develop are excised from calli and transplanted to appropriate root-inducing selective medium.","Rooted plantlets are transplanted to soil as soon as possible after roots appear.","The plantlets can be repotted as required, until reaching maturity.","In vegetatively propagated crops, the mature tralsgenic plants are propagated utilizing cuttings or tissue culture techniques to produce multiple identical plants.","Selection of desirable transgenotes is made and new varieties are obtained and propagated vegetatively for commercial use.","In seed propagated crops, the mature transgenic plants can be self-pollinated to produce a homozygous inbred plant.","The resulting inbred plant produces seeds that contain the introduced transgene, which can comprise a heterologous nucleotide sequence encoding a bioluminescent polypeptide, and can be grown to produce plants that express the polypeptide.","As such, the invention provides seeds produced by a transgenic plant obtained by a method of the invention.","Transgenic plants comprising different transgenes also can be crossbred, thereby providing a means to obtain transgenic plants containing two or more different transgenes, one of which encodes a bioluminescent polypeptide and the other or others of which confer a desirable characteristic to the plant.","Methods for breeding plants and selecting for crossbred plants having desirable characteristics or other characteristics of interest are well known in the art.","A method of the invention can be performed by introducing a recombinant nucleic acid molecule of the invention into a plant cell.","As used herein, the term “introducing” means transferring a polynucleotide into a plant cell.","A nucleic acid molecule can be introduced into a cell by a variety of methods.","For example, the recombinant nucleic acid molecule of the invention, which can be contained in a vector, can be introduced into a plant cell using a direct gene transfer method such as electroporation or microprojectile mediated transformation, or using Agrobacterium mediated transformation.","As used herein, the term “transformed” refers to a plant cell containing an exogenously introduced nucleic acid molecule.","It should be recognized that one or more nucleic acid molecules, which are the same or different and at least one of which is a recombinant nucleic acid molecule of the invention, can be introduced into a plant cell, thereby providing a means to obtain a genetically modified plant cell containing multiple copies of a single transgene, or containing two or more different genes, either or both of which can be present in multiple copies.","Such genetically modified plant cells can be produced, for example, by simply selecting plant cells having multiple copies of a single type of transgene; by cotransfecting plant cells with two or more populations of different transgenes and identifying those containing the two or more different transgenes; or by crossbreeding transgenic plants, each of which contains one or more desired transgenes, and identifying those progeny having the desired transgenes.","Methods for introducing a nucleic acid molecule into a plant cell to obtain a transformed plant also include direct gene transfer (see European Patent A 164 575), injection, electroporation, biolistic methods such as particle bombardment, pollen-mediated transformation, plant RNA virus-mediated transformation, liposome-mediated transformation, transformation using wounded or enzyme-degraded immature embryos, or wounded or enzyme-degraded embryogenic callus, and the like.","Transformation methods using Agrobacterium tumefaciens tumor inducing (Ti) plasmids or A. rhizogenes root-inducing (Ri) plasmids, or other plant virus vectors are well known in the art (see, for example, WO 99/47552; Weissbach and Weissbach, “Methods for Plant Molecular Biology” (Academic Press, NY 1988), section VIII, pages 421-463; Grierson and Corey, “Plant Molecular Biology” 2d Ed.","(Blackie, London 1988), Chapters 7-9, each of which is incorporated herein by reference; Horsch et al., supra, 1985).","The wild type form of A. tumefaciens, for example, contains a Ti plasmid, which directs production of tumorigenic crown gall growth on host plants.","Transfer of the tumor inducing T-DNA region of the Ti plasmid to a plant genome requires the Ti plasmid-encoded virulence genes as well as T-DNA borders, which are a set of direct DNA repeats that delineate the region to be transferred.","An Agrobacterium based vector is a modified form of a Ti plasmid, in which the tumor inducing functions are replaced by a nucleotide sequence of interest that is to be introduced into the plant host.","Methods of using Agrobacterium mediated transformation include cocultivation of Agrobacterium with cultured isolated protoplasts; transformation of plant cells or tissues with Agrobacterium; and transformation of seeds, apices or meristems with Agrobacterium.","In addition, in planta transformation by Agrobacterium can be performed using vacuum infiltration of a suspension of Agrobacterium cells (Bechtold et al., C.R.","Acad.","Sci.","Paris 316:1194, 1993, which is incorporated herein by reference).","Agrobacterium mediated transformation can employ cointegrate vectors or binary vector systems, in which the components of the Ti plasmid are divided between a helper vector, which resides permanently in the Agrobacterium host and carries the virulence genes, and a shuttle vector, which contains the gene of interest bounded by T-DNA sequences.","Binary vectors are well known in the art (see, for example, De Framond, BioTechnology 1:262, 1983; Hoekema et al., Nature 303:179, 1983, each of which is incorporated herein by reference) and are commercially available (Clontech; Palo Alto Calif.).","For transformation, Agrobacterium can be cocultured, for example, with plant cells or wounded tissue such as leaf tissue, root explants, hypocotyls, cotyledons, stem pieces or tubers (see, for example, Glick and Thompson, “Methods in Plant Molecular Biology and Biotechnology” (Boca Raton Fla., CRC Press-1993), which is incorporated herein by reference).","Wounded cells within the plant tissue that have been infected by Agrobacterium can develop organs de novo when cultured under the appropriate conditions; the resulting transgenic shoots eventually give rise to transgenic plants, which contain an exogenously introduced circadian-regulated polynucleotide, or an oligonucleotide portion thereof.","Agrobacterium mediated transformation has been used to produce a variety of tratsgenic plants, including, for example, transgenic cruciferous plants such as Arabidopsis, mustard, rapeseed and flax; transgenic leguminous plants such as alfalfa, pea, soybean, trefoil and white clover; and transgenic solanaceous plants such as eggplant, petunia, potato, tobacco and tomato (see, for example, Wang et al., “Transformation of Plants and Soil Microorganisms” (Cambridge, University Press 1995), which is incorporated herein by reference).","In addition, Agrobacterium mediated transformation can be used to introduce an exogenous nucleic acid molecule into apple, aspen, belladonna, black currant, carrot, celery, cotton, cucumber, grape, horseradish, lettuce, morning glory, muskmelon, neem, poplar, strawberry, sugar beet, sunflower, walnut, asparagus, rice and other plants (see, for example, Glick and Thompson, supra, 1993; Hiei et al., Plant J.","6:271-282, 1994; Shimamoto, Science 270:1772-1773, 1995).","Suitable strains of A. tumefaciens and vectors as well as transformation of Agrobacteria and appropriate growth and selection media are well known in the art (GV3101, pMK90RK), Koncz, Mol.","Gen. Genet.","204:383-396, 1986; (C58C1, pGV3850kan), Deblaere, Nucl.","Acid Res.","13:4777, 1985; Bevan, Nucleic Acid Res.","12:8711, 1984; Koncz, Proc.","Natl.","Acad.","Sci.","USA 86:8467-8471, 1986; Koncz, Plant Mol.","Biol.","20:963-976, 1992; Koncz, Specialized vectors for gene tagging and expression studies.","In: Plant Molecular Biology Manual Vol.","2, Gelvin and Schilperoort (Eds.","), Dordrecht, The Netherlands: Kluwer Academic Publ.","(1994), 1-22; European Patent A-1 20 516; Hoekema: The Binary Plant Vector System, Offsetdruikkerij Kanters B. V., Alblasserdam (1985), Chapter V; Fraley, Crit.","Rev.","Plant.","Sci., 4:1-46; An, EMBO J.","4:277-287, 1985).","Where the recombinant nucleic acid molecule is contained in a vector, the vector can contain functional elements, for example “left border” and “right border” sequences of the T-DNA of Agrobacterium, which allow for stable integration into a plant genome.","Furthermore, methods and vectors that permit the generation of marker-free transgenic plants, for example, where a selectable marker gene is lost at a certain stage of plant development or plant breeding, are known, and include, for example, methods of co-transformation (Lyznik, Plant Mol.","Biol.","13:151-161, 1989; Peng, Plant Mol.","Biol.","27:91-104, 1995), or methods that utilize enzymes capable of promoting homologous recombination in plants (see, e.g., W097/08331; Bayley, Plant Mol.","Biol.","18:353-361, 1992; Lloyd, Mol.","Gen. Genet.","242:653-657, 1994; Maeser, Mol.","Gen. Genet.","230:170-176, 1991; Onouchi, Nucl.","Acids Res.","19:6373-6378, 1991; see, also, Sambrook et al., supra, 1989).","A direct gene transfer method such as electroporation also can be used to introduce a polynucleotide into a cell such as a plant cell.","For example, plant protoplasts can be electroporated in the presence of a recombinant nucleic acid molecule comprising a nucleotide sequence encoding a bioluminescent polypeptide, which can be in a vector (Fromm et al., Proc.","Natl.","Acad.","Sci., USA 82:5824, 1985, which is incorporated herein by reference).","Electrical impulses of high field strength reversibly permeabilize membranes allowing the introduction of the nucleic acid.","Electroporated plant protoplasts reform the cell wall, divide and form a plant callus.","Microinjection can be performed as described in Potrykus and Spangenberg (eds.","), Gene Transfer To Plants.","Springer Verlag, Berlin, N.Y. (1995).","A transformed plant cell containing the introduced recombinant nucleic acid molecule can be identified due to the presence of a selectable marker included in the construct, or simply by looking at the plant cells and observing visible bioluminescence.","Microprojectile mediated transformation also can be used to introduce a recombinant nucleic acid molecule of the invention into a plant cell (Klein et al., Nature 327:70-73, 1987, which is incorporated herein by reference).","This method utilizes microprojectiles such as gold or tungsten, which are coated with the desired nucleic acid molecule by precipitation with calcium chloride, spermidine or polyethylene glycol.","The microprojectile particles are accelerated at high speed into a plant tissue using a device such as the BIOLISTIC PD-1000 particle gun (BioRad; Hercules Calif.).","Microprojectile mediated delivery (“particle bombardment”) is especially useful to transform plant cells that are difficult to transform or regenerate using other methods.","Methods for the transformation using biolistic methods are well known (Wan, Plant Physiol.","104:37-48, 1984; Vasil, BioTechnology 11:1553-1558, 1993; Christou, Trends in Plant Science 1:423-431, 1996).","Microprojectile mediated transformation has been used, for example, to generate a variety of transgenic plant species, including cotton, tobacco, corn, hybrid poplar and papaya (see Glick and Thompson, supra, 1993).","Important cereal crops such as wheat, oat, barley, sorghum and rice also have been transformed using microprojectile mediated delivery (Duan et al., Nature Biotech.","14:494-498, 1996; Shimamoto, Curr.","Opin.","Biotech.","5:158-162, 1994).","A rapid transformation regeneration system for the production of transgenic plants such as a system that produces transgenic wheat in two to three months (see European Patent No.","EP 0709462A2, which is incorporated herein by reference) also can be useful for producing a transgenic plant according to a method of the invention, thus allowing more rapid identification of gene functions.","The transformation of most dicotyledonous plants is possible with the methods described above.","Transformation of monocotyledonous plants also can be transformed using, for example, biolistic methods as described above, protoplast transformation, electroporation of partially permeabilized cells, introduction of DNA using glass fibers, Agrobacterium mediated transformation, and the like.","Plastid transformation also can be used to introduce a nucleic acid molecule into a plant cell (U.S. Pat.","Nos.","5,451,513, 5,545,817, and 5,545,818; WO 95/16783; McBride et al., Proc.","Natl.","Acad.","Sci., USA 91:7301-7305, 1994).","Chloroplast transformation involves introducing regions of cloned plastid DNA flanking a desired nucleotide sequence, for example, a selectable marker together with polynucleotide of interest into a suitable target tissue, using, for example, a biolistic or protoplast transformation method (e.g., calcium chloride or PEG mediated transformation).","One to 1.5 kb flanking regions (“targeting sequences”) facilitate homologous recombination with the plastid genome, and allow the replacement or modification of specific regions of the plastome.","Using this method, point mutations in the chloroplast 16S rRNA and rps12 genes, which confer resistance to spectinomycin and streptomycin and can be utilized as selectable markers for transformation (Svab et al., Proc.","Natl.","Acad.","Sci., USA 87:8526-8530, 1990; Staub and Maliga, Plant Cell 4:39-45, 1992), resulted in stable homopiasmic transformants, at a frequency of approximately one per 100 bombardments of target leaves.","The presence of cloning sites between these markers allowed creation of a plastid targeting vector for introduction of foreign genes (Staub and Maliga, EMBO J.","12:601-606, 1993).","Substantial increases in transformation frequency are obtained by replacement of the recessive rRNA or r-protein antibiotic resistance genes with a dominant selectable marker, the bacterial aadA gene encoding the spectinomycin-detoxifying enzyme aminoglycoside-3′-adenyltransferase (Svab and Maliga, Proc.","Natl.","Acad.","Sci., USA 90:913-917, 1993).","Approximately 15 to 20 cell division cycles following transformation are generally required to reach a homoplastidic state.","Plastid expression, in which genes are inserted by homologous recombination into all of the several thousand copies of the circular plastid genome present in each plant cell, takes advantage of the enormous copy number advantage over nuclear-expressed genes to permit expression levels that can readily exceed 10% of the total soluble plant protein.","Plants suitable for treatment according to a method of the invention so as to be visibly bioluminescent can be monocots or dicots and include, but are not limited to, maize, wheat, barley, rye, sweet potato, bean, pea, chicory, lettuce, cabbage, cauliflower, broccoli, turnip, radish, spinach, asparagus, onion, garlic, pepper, celery, squash, pumpkin, hemp, zucchini, apple, pear, quince, melon, plum, cherry, peach, nectarine, apricot, strawberry, grape, raspberry, blackberry, pineapple, avocado, papaya, mango, banana, soybean, tomato, sorghum, sugarcane, sugar beet, sunflower, rapeseed, clover, tobacco, carrot, cotton, alfalfa, rice, potato, eggplant, cucumber, Arabidopsis thaliana, and woody plants such as coniferous and deciduous trees.","Thus, a transgenic plant or genetically modified plant cell of the invention can be an angiosperm or gymnosperm.","Angiosperms are divided into two broad classes based on the number of cotyledons, which are seed leaves that generally store or absorb food; a monocotyledonous angiosperm has a single cotyledon, and a dicotyledonous angiosperm has two cotyledons.","Angiosperms produce a variety of useful products including materials such as lumber, rubber, and paper; fibers such as cotton and linen; herbs and medicines such as quinine and vinblastine; ornamental flowers such as roses and, where included within the scope of the present invention, orchids; and foodstuffs such as grains, oils, fruits and vegetables.","Angiosperms encompass a variety of flowering plants, including, for example, cereal plants, leguminous plants, oilseed plants, hardwood trees, fruit-bearing plants and ornamental flowers, which general classes are not necessarily exclusive.","Cereal plants, which produce an edible grain cereal, include, for example, corn, rice, wheat, barley, oat, rye, orchardgrass, guinea grass, sorghum and turfgrass.","Leguminous plants include members of the pea family (Fabaceae) and produce a characteristic fruit known as a legume.","Examples of leguminous plants include, for example, soybean, pea, chickpea, moth bean, broad bean, kidney bean, lima bean, lentil, cowpea, dry bean, and peanut, as well as alfalfa, birdsfoot trefoil, clover and sainfoin.","Oilseed plants, which have seeds that are useful as a source of oil, include soybean, sunflower, rapeseed (canola) and cottonseed.","Angiosperms also include hardwood trees, which are perennial woody plants that generally have a single stem (trunk).","Examples of such trees include alder, ash, aspen, basswood (linden), beech, birch, cherry, cottonwood, eln, eucalyptus, hickory, locust, maple, oak, persimmon, poplar, sycamore, walnut, sequoia, and willow.","Trees are useful, for example, as a source of pulp, paper, structural material and fuel.","Angiosperms are fruit-bearing plants that produce a mature, ripened ovary, which generally contains seeds.","A fruit can be suitable for human or animal consumption or for collection of seeds to propagate the species.","For example, hops are a member of the mulberry family that are prized for their flavoring in malt liquor.","Fruit-bearing angiosperms also include grape, orange, lemon, grapefruit, avocado, date, peach, cherry, olive, plum, coconut, apple and pear trees and blackberry, blueberry, raspberry, strawberry, pineapple, tomato, cucumber and eggplant plants.","An ornamental flower is an angiosperm cultivated for its decorative flower.","Examples of commercially important ornamental flowers include rose, lily, tulip and chrysanthemum, snapdragon, camellia, carnation and petunia plants, and can include orchids.","The skilled artisan will recognize that the methods of the invention can be practiced using these or other angiosperms, as desired, as well as gymnosperms, which do not produce seeds in a fruit.","As such, the methods of the invention can be used to prepare bioluminescent plants of various families, including, for example, Asteraceae-Aster family; Fabaceae-Pea family; Rubiaceae-Madder family; Poaceae-Grass family; Euphorbiaceae-Spurge family; Lamiaceae-Mint family, Melastomataceae-Melastome family; Liliaceae-Lily family; Scrophulariaceae-Figwort family; Acanthaceae-Ancanthus family, Myaceae-Myrtle family, Cyperaceae-Sedge family; Ericaceae-Heath family, Apiaceae-Carrot family, Rosaceae-Rose family; Brassicaceae-Mustard family, Araceae-Arum family, Asclepiadaceae-Milkweed family; Arecaceae-Pahm family, Solanaceae-Potato family, Boraginaceae-Borage family; Gesneriaceae-Generiad family, Lauraceae-Laurel family, and Bromeliaceae-Bromeliad family.","In certain embodiments, the methods of the invention also can be used to prepare bioluminescent plants of Orchidaceae-Orchid family, whereas in other embodiments, members of the orchid family are specifically excluded.","A recombinant nucleic acid molecule of the invention or a vector containing the recombinant nucleic acid molecule provides a valuable reagent for identifying plant cells containing an introduced nucleic acid molecule.","For example, the recombinant nucleic acid molecule, or vector containing the recombinant nucleic acid molecule, can include a nucleotide sequence of interest.","Following transformation of plant cells with the recombinant nucleic acid molecule of the invention, which contains the nucleotide sequence of interest, visibly bioluminescent plant cells can be selected simply by looking at the plant cells, for example, under a microscope, and picking out the ones that are visibly bioluminescent, thereby selecting plant cells that also contain the nucleotide sequence of interest.","Thus, a recombinant nucleic acid molecule of the invention is useful as a selectable marker, and provides a quick, efficient, and inexpensive means to identify a plant cell containing a nucleotide sequence of interest.","A nucleotide sequence of interest also can encode an immunogenic polypeptide, for example, a viral or bacterial polypeptide, or a peptide portion thereof.","A genetically modified plant cell, particularly a transgenic plant comprising such a plant cell, can be used as a source of a vaccine, wherein the individuals to be immunized merely ingest the plant, and a protective or palliative immune response is obtained.","As an additional advantage, problems generally associated with the preparation and purification of a vaccine, as well as storage and refrigeration problems and the like are obviated.","In view of these examples, it will be recognized that genetically modified plant cells and transgenic plant comprising such cells containing essentially any nucleotide sequence of interest can be prepared, and that the inclusion of a recombinant nucleic acid molecule of the invention provides a selectable marker that facilitates the identification of a transformed plant cell or plant comprising such a cell that contains the nucleotide sequence of interest A recombinant nucleic acid molecule of the invention, or a vector containing such a nucleic acid molecule, also is useful for preparing transgenic plants having great ornamental value.","Visibly bioluminescent plants containing genetically modified plant cells have not previously been described.","As such, the transgenic plants of the invention, which “glow in the dark,” provide a novel ornamental plant.","For example, a transgenic plant expressing a luciferase polypeptide or variant thereof can be sprayed with water containing luciferin, or the transgenic plant or a cutting or other portion of the transgenic plant, for example, an inflorescence of the plant, can be dipped or placed in a solution containing luciferin, whereupon the visible bioluminescence is produced.","For example, flowers can be cut from a transgenic plant of the invention and placed in water containing luciferin, wherein the luciferin is taken up and transported to the flower, thus providing a visibly bioluminescent flower.","Generally, the amount of luciferin sufficient to induce visible bioluminescence includes at least about 0.1 mM, and concentrations from about 0.1 mM to 5 mM or more can be used.","In addition, the solution containing luciferin can further include a surfactant, for example, a non-ionic detergent such as TRITON X-100 detergent or NONIDET P40 detergent in a concentration of about 0.001% to about 0.1%, thus facilitating entry of the luciferin into the plant cells.","Significantly, luciferase-mediated bioluminescence continues for several hours following a single administration of luciferin and, therefore, there is no requirement that the plants be continually treated to maintain bioluminescence.","Furthermore, the transgenic plants can be selected based on their expression of variable levels of light, including plants that glow dimly, yet visibly, and those that glow very brightly.","In addition to providing general ornamental value, the visibly bioluminescent transgenic plants of the invention also can provide a practical value.","For example, visibly bioluminescent transgenic plants that grow as bushes, shrubs or trees can be arranged along a walkway or driveway.","Where such plants express luciferase or a variant thereof, for example, the plants can be watered in the early evening with a solution containing luciferin such that they illuminate the walkway or driveway after night falls.","Thus, the transgenic plants of the invention provide the additional advantage they can be used to make an area safer by illuminating the area.","The present invention also relates to kit, which contains a genetically modified plant cell of the invention, or a derivative of the genetically modified plant cell, for example, a transgenic plant derived from the genetically modified plant cell, or a cell, a tissue, or an organ of such a transgenic plant.","As such, a kit of the invention can contain one or more flowers, bracts, leaves or other tissues or organs, which, upon contact with luciferin, are visibly luminescent.","Accordingly, a kit of the invention can further include an amount of luciferin sufficient for generating visible bioluminescence of the genetically modified plant cell or the derivative of the genetically modified plant cell, and also can include a plurality of such amounts of luciferin, thus allowing the generation of visible bioluminesce a number of times, as desired.","In addition, a kit of the invention can include an amount of a carotenoid inhibitor, for example, norfluorazon, sufficient for reducing or inhibiting chlorophyll production in the genetically modified plant cell or the derivative of the genetically modified plant cell.","In another embodiment, a kit of the invention contains one or more cuttings, seeds, or other portion or derivative of a visibly bioluminescent plant of the invention such that a visibly luminescent transgenic plant can be grown therefrom.","As such, the kit also can include reagents for growing a transgenic plant from the cutting or seed, including, for example, a suitable plant food or other nutrient source required for growth of the particular plant.","In addition, the kit can include an amount of a carotenoid inhibitor sufficient for reducing or inhibiting chlorophyll production in a transgenic plant grown from the cutting or seed; and/or can include an amount of luciferin sufficient for generating visible bioluminescence of a transgenic plant grown from the cutting or seed.","Generally, a kit of the invention includes a container for the components of the kit.","It should be recognized, however, that the container can be any convenient means for holding a component of the kit, including, for example, a packet or collection of packets containing seeds obtained from a transgenic plant of the invention, or containing predetermined amounts of a reagent such as luciferin, which can be added to a specified amount of water to provide a solution sufficient to generate visible bioluminescence from a composition of the invention.","In one embodiment, the kit comprises a flower cutting of a transgenic plant of the invention in a vase, and can further include one or more predetermined amounts of a reagent such as luciferin.","In another embodiment, the kit comprises a seed or a cutting obtained from a transgenic plant of the invention, and further includes a reagent such as luciferin and/or norfluorazon, in which the seed or cutting, or a plant derived therefrom, can be grown.","The following examples are intended to illustrate but not limit the invention.","EXAMPLE 1 Preparation of Luciferase Expression Vectors This example provides a method for preparing luciferase constructs that can be incorporated into an expression vector and are useful for producing a visibly bioluminescent plant.","Plasmid pRTL2, which contains a CaMV 35S promoter with duplicated enhancer, a TEV leader and initial coding sequence from pTL-7SN, and a CaMV 35S polyadenylation (poly A) signal sequence (Topfer et al., Nucl.","Acids Res.","15:5890, 1987; Restrepo et al., Plant Cell 2:987, 1990, each of which is incorporated herein by reference; see, also, Carrington and Freed, supra, 1990) was sequentially digested with Nco I and Xba I, then separated by electrophoresis on a 0.8% agarose gel.","The nucleotide sequence of the CaMV 35S promoter with duplicated enhancer, TEV leader and initial coding sequence, and CaMV 35S poly A signal sequence of pRTL2 is shown as SEQ ID NO:3.A DNA band of approximately 3.9 kb representing the linear pRTL2 (see nucleotides 1 to 1188 and 2844 to 5475 of SEQ ID NO:1) was isolated and extracted from the gel using a QIAQUICK DNA gel extraction kit (Qiagen; Valencia Calif.) according to manufacturer's instructions.","The Nco I site in the TEV leader contains the start codon for translation.","Translational fusions containing the initial 20 amino acid residues of the TEV polyprotein are made by ligation to one of the polylinker sites, provided care is taken to avoid the STOP codon in the Xba I site (see SEQ ID NO:3).","In a parallel reaction, pGL3 (Promega Corp., Madison Wis.), which contains a modified luciferase coding sequence, luc+ (Groskreutz et al., Promega Notes 50:2, 1995; U.S. Pat.","No.5,670,356, each of which is incorporated herein by reference) was digested sequentially with Xba I and Nco I, then separated by electrophoresis on a 0.8% agarose gel.","Two bands were present—a 1.6 kb band representing the luc+ coding sequence (see U.S. Pat.","No.5,670,356) and an approximately 3 kb band representing the remainder of the pGL3 plasmid.","The approximately 1.6 kb luc+ band (see nucleotides 1188 to 2844of SEQ ID NO:1) was isolated and extracted as described above.","The isolated luc+ sequence (4 μl) was ligated into the linearized pRTL2 vector (1 μl) using ligase and ligation buffer from BRL (Gaithersburg Md.).","The ligation was performed overnight at 16° C. E. coli DH5α cells were transformed using a BioRad Electroporator in a ratio of 2 μl of ligated DNA to 20 μl of cells.","Following electroporation, the transformed cells were incubated in LB medium for 1 hr at 37° C., then were plated on LB plates containing 50 μg/ml ampicillin.","Of approximately 3000 colonies, 32 were selected and screened by PCR for the luc+ insert.","PCR primers were specific for the 5′ end of the TEV promoter in pRTL2 and for the 3′ end of the poly A terminator in pRTL2 (see SEQ ID NO:3).","PCR analysis revealed that 30 of the colonies contained the luc+ insert.","The presence of the insert in 6 clones was verified by restriction analysis, and two candidate clones, designated TK1-7 and TK1-8, were sequenced to confirm the presence of the luc+ insert.","The TK1-8 clone was digested at 37° C. for 2 hr with Hind III, and the fragment containing the CaMV 35S promoter with duplicated enhancer, TEV leader, luc+ coding sequence and poly A termination signal (designated “TK1829”; see FIG.","2, and nucleotides 8366 to 11,113 of SEQ ID NO:2) was isolated by electrophoresis as described above.","The binary vector, pPZP221, also was digested with Hind III and isolated.","The linearized pPZP221 vector was treated with shrimp alkaline phosphatase for 1 hr at 37° C., to remove terminal phosphate groups, followed by 15 min at 65° C. to heat inactivate the enzyme, then the TK1829 insert and linearized pPZP221 vector were ligated and transformed into DH5α cells as described above.","Cells were plated on LB agar plates containing X-gal, IPTG, and 100 μg/ml spectinomycin.","Forty-six white colonies were selected, and the presence of the luc+ coding sequence was confirmed by PCR as described above; all were positive for the luc+ sequence.","Six samples were selected and the orientation of the insert was determined by restriction digest analysis, using Sca I or Nar I.","One construct (#29) having the correct orientation was selected.","The binary vector containing the luc+ insert was designated pPZPTK1829 (FIG.","2; SEQ ID NO:2), and the inserted expression construct was designated TK1829 (nucleotides 8366 to 11,113 of SEQ ID NO:2).","TK1829 was sequenced using primers specific for 5′ and 3′ insert flanking regions of the pPZP221 vector.","The pRTL2 vector from which the TK1829 insert was obtained was designated pTK1829 (see FIG.","1; see, also, SEQ ID NO:1).","The pMT-OM-LUC+ (SEQ ID NO:4; FIG.","3) and pACT-OM-LUC+ (SEQ ID NO:5; FIG.","4) binary vector plasmids were prepared from the pPZP221 binary vector, as described above.","The metallothionein (MT) or actin (ACT) promoter was inserted immediately preceding the omega enhancer in a Bam HI site, which provides a convenient site to substitute other transcriptional regulatory elements.","EXAMPLE 2 Preparation and Characterization of Visibly Bioluminescent Plants This example provides a method for introducing a luciferase expression construct into plants such that a genetically modified visibly bioluminescent plant is produced, and a method for determining a threshold value of photon emission required for visible luminescence.","A.","Preparation of TK1829 Transformed Plants The TK1829 expression construct was introduced from pPZPTK1829 into Agrobacterium l strain ABI 1 with the help of pRK2013 by tri-parental mating.","Agrobacterium containing the TK1829 construct were selected based on their resistance to the 100 μg/ml spectinomycin, 50 μg/ml kanamycin, and 25 μg/ml chloramphenicol.","TK1829-Agrobacterium strains were grown, then DNA was purified and checked by restriction digest analysis using Sca I (2 hr; 37° C.) to verify the presence of the insert.","Briefly, TK1829-Agrobacterium was grown as described below, and 28 day old plants were infiltrated.","Supernova (Snv) strain plants and Columbia (wild type) plants were each infiltrated.","Two sets of infiltrations were performed on Columbia and Snv plants.","Each time, two 4 inch pots of each line were used and each pot contained approximately 50 plants, for a total of 200 plants per line infiltrated.","For in planta plant transformation, 4 inch pots were prepared with Scott's Metro mix 200 soil and covered with window screen mesh.","Soil was wet and mounded up, and the screen mesh was secured by wrapping a rubber band around the neck of the pot, maling sure that the mesh was in contact with the soil surface so that germinating seedlings could get through the mesh.","Seeds were stratified in 10 to 50 ml of sterile water and stored at 4° C. for 3 days prior to being applied to pots.","Seeds were sown using a dropper, to spread seeds evenly over the surface of the mesh.","Plants were grown at approximately 22° C., in 12 hr light:12 hr dark per day to a stage at which bolts were anywhere from 3 to 10 inches high.","The bolts were cut off to induce growth of secondary inflorescence.","Infiltration was most effective 4 to 5 days after decapitation of bolts, but can be performed up to 8 days later.","For infiltration, 5 to 20 ml liquid preculture of Agrobactenium containing the appropriate construct were grown for about 2 days in a shaking incubator at 28° C., then 400 ml of LB was inoculated with the preculture and incubate for one additional day.","The large culture generally was prepared the day before the infiltration was performed.","Transformed Agrobacterium cells were grown to an optical density at 600 nm (OD600) of approximately 1.2, then harvested by centrifugation (4000 rpm, 10 min, room temperature), and resuspended in infiltration medium (1/2×Murashige and Skoog salts; 1×Gamborg's vitamins; 5.0% sucrose; 0.44 μM benzylamino-purine (10 μg per liter of a 1 ng/ml stock); pH adjusted to 5.7 using KOH; then add 200 μl/l Silwet; media need not be sterilized if used immediately) to an OD 600 of approximately 0.8.The OD600 of an overnight culture generally was about 1.2 to 1.6 units.","A 400 ml suspension generally was adequate for an infiltration procedure.","The same suspension can be used as many as three times.","About 150-200 ml of the Agrobacterium suspension was added to either a beaker or dish and the prepared plants were inverted into the suspension.","The plants were entirely submerged and kept in the solution for 10 to 15 min.","The pots then were removed from the suspension and drained thoroughly, laid on their side in a plastic flat, and covered with a plastic dome or Saran Wrap to maintain humidity.","The flat was uncovered the next day and the pots set upright.","A little water was added to the bottom of the tray to help keep the humidity level high while the plants were recovering.","The plants were grown for 3 to 4 weeks, keeping the bolts from each pot together and separated from neighboring pots and different infiltrations.","Once plants began to yellow and senesce, watering was reduced or discontinued.","Seeds were harvested together from one pot.","Transformants were selected on 150×15mm selection plates (1/2×Murashige and Skoog salts; 1×Gamborg's vitamins; 1% sucrose; 0.5 g/l MES; 0.8% agarose; pH to 5.7; autoclaved, then add 75 μg/ml gentamycin, 500 μg/ml carbenicilin, 50 μg/ml kanamycin).","Seeds were surface sterilize for 2 min in 500 μl 70% ethanol; followed by 10 min in 40% bleach, 1% SDS, 59% water, then rinsed 3 times with sterile water.","The seeds were resuspended in 3 to 4 ml of sterile water and plated by pouring onto a plate and shaking until evenly distributed.","The plates were allowed to sit for 5 to 10 min, then excess liquid was removed.","Approximately 70 to 100 μl of dry seeds were added per plate.","Plants were allowed to stratify at 4° C. for 3 days, then transferred to a growth chamber.","In some experiments, the herbicide, norflurazon (5 μM; 4-chloro-5-(methylamino)-2-{3-(tri-fluoromethyl)phenyl}-3(2H)-pyridazinone) was included in the growth medium.","Norflurazon inhibits chlorophyll synthesis, resulting in the growth of “white” plants.","Treatment of plants with norflurazon resulted in greater emission of visible bioluminescence due to the lack of chlorophyll, which absorbs light and blocks light emission (see Tables).","After 7 to 10 days (depending on the antibiotic resistance selected for), transformants were identified as dark green plants with elongated roots that penetrated the agar.","These seedlings were transferred onto a second set of selection plates.","After several days, plants had 4 to 8 true leaves and branching roots.","Pseudo-resistant plants were unable to survive in this medium.","The plants then were transferred to soil, and kept covered for the first few days.","Plants were allowed to recover, bolt, set seed and senesce.","Seeds were harvested when dry, then were further dried in a dessicator for 1 week.","Yield was approximately 0.5 to 0.75 ml of seed per pot.","T1 (transgenic first generation) seeds in the Columbia background were screened on 75 μg/ml gentamycin and 500 μg/ml carbenicilin MS plates.","T1 seeds in the Snv background were screened on 75 μg/ml gentamycin, 500 μg/ml carbenicilin, and 50 μg/ml kanamycin (Snv strain contains a transgene conferring kanamycin resistance).","Over an 8 week period, 1.6 ml (approximately 32,000 seeds) of each T1 seed set was sterilized and planted in 200 μl (4000 seeds) aliquots on 50 ml plates as described above.","Plants were stratified for 3 days at 4° C., then moved to a growth chamber set for 12 hr light: 12 hr dark and approximately 27° C. Sixty-seven T1 seedlings in the Snv background and 25 T1 seedlings in the Columbia background grew under the selection conditions, and were recovered and transferred to soil to harvest T2 seed.","It took approximately 12 to 16 weeks to take T1 seed to T2 seed, and varied between plants.","B.","Characterization of TK1829 Transformed Plants T2 seeds of Columbia and Supernova were sterilized and plated as above, and screened for antibiotic resistance.","In addition, the seedlings were screened for bioluminescence, visually and with the Hamamatsu photon counting camera.","T2 seedlings within individual families were selected based on their intensity and brightness.","Three lines in the Snv background (Snv 11, Snv 37 and Snv 57) and two lines in the Columbia background (Col 10 and Col 19) were selected as the most visibly bioluminescent plants.","Seventy-eight T3 individuals of Snv 11, which was extremely bright, were selected based on their visible bioluminescence.","Three of these (Snv 11-8, Snv 11-48, Snv 11-52) were selected as the brightest and were maintained as individual lines.","For characterization of luciferase expression, tissue was collected from 22 day old plants grown under the same conditions.","Examination of Columbia wild-type plants, Columbia plants transformed with 35S:Luc (“35S:Luc”) and Snv plants transformed with cab2:Luc (“Supernova”; see Hicks et al., supra, 1996) was included for comparison.","In various assays (see below), protein concentration was determined, relative light units (RLU) were acquired using a luminometer, western blot analysis was performed using recombinant luciferase antibodies, plants were imaged using a Night Owl Camera (see below) to obtain counts of light/mm2/sec, and plants were inspected visually to confirm that bioluminescence was visible to the eye.","Based on these assays, the approximate amounts of luciferase per microgram of total protein and the fold increase in enzyme activity were determined, and the amount of light emitted by the plants was determined.","For protein assays, a few hundred 22 day old seedlings from plants grown under the same conditions were placed in liquid nitrogen, ground into a fine power, and stored in aliquots of approximately 200 μl volume on dry ice or at −80° C. until the time proteins were extracted.","Approximately 100 mg of the powdered tissue was extracted in 500 μl of 1×Promega Cell Lysis Reagent (CLR), ground briefly with a mortar and pestle, then centrifuged at 4° C. for 5 min 14,000 rpm.","The supernatant was divided into 100 μl aliquots and transferred immediately to dry ice.","Any samples not used within 1 hr were stored at −80° C. to preserve enzyme activity.","Protein concentrations were determined for 20 μl of tissue extract using the DC Protein Assay Reagent (BioRad); optical density (OD) was determined at 750 nm using a Pharmacia Ultrospec III spectrophotometer.","Acquired Relative Light Units (RLU) were obtained using a Turner TD20E Luminometer.","Samples from plants containing the TK1829 construct were diluted 1/100 (5 μl/500 μl) in order for the reading to be within the range of the luminometer.","Five μl of sample and 50 μl of Promega Luciferase Assay Reagent (LAR) was used.","The maximum value of the range of readings taken every 30 sec for the first 5 min of the reaction was entered into the data tables.","The amount of luciferase was approximated by western blot analysis (Tables 1 and 2) or using the RLU data obtained by luminometry and the total protein concentration obtained by spectrophotometry (Table 4).","Western blot analysis of the extracts was performed using enhanced chemifluoresence (ECF; Amersham) to visualize protein bands.","For samples containing the TK1829 construct, 0.1 μg of total protein was used.","For all other samples, 1.0 μg of total protein was used.","The 10-fold difference was necessary to keep the luciferase protein concentrations within the range of the assay.","A luciferase primary antibody and an alkaline phosphatase secondary antibody (Promega) were used to identify protein bands.","Proteins were separated by electrophoresis on, 10% SDS PAGE gels (BioRad), then transferred to Hybond nitrocellulose.","Images were visualized using a Molecular Dynamics Fluorimeter, and data was analyzed using ImageQuant software.","Luciferase was expressed in 35S:Luc transformed Columbia plants (“35S:Luc”) and cab2:Luc transformed Supernova plants (“Supernova”; Table 1), but these plants were not visibly bioluminescent when examined by eye.","In comparison, the TK1829 transformed Columbia plants were visibly bioluminescent.","These results indicate that the threshold for visible bioluminescence is between about 85 and 125 pg luc+ protein/μg protein as determined by western blot analysis (Table 1).","The TK1829 transformed Supernova plants expressed about 6.5-fold more luc+ than the TK1829 transformed Columbia plants as determined by western blot analysis (Table 1).","In addition, the Snv-TK1829 plants emitted about 7-fold more photons of light/mm2/sec than the Col-TK1829 plants.","These results demonstrate that visible bioluminescence can be produced in various Arabidopsis strains.","In addition, growth of the TK1829 transformed Supernova plants in norflurazon resulted in an almost 2-fold increase in luciferase protein as measured by luminometry, and a greater than 7-fold amount of light emitted as measured using the Night Owl imaging system.","This result demonstrates that the amount of visible bioluminescence can be increased by inhibiting chlorophyll synthesis in the transgenic plants.","TABLE 1 pg Luciferase Plant Line protein/μg total protein a) Columbia Wild Type 0.01 b) 35S:Luc transformed Columbia 7.0 c) cab2:Luc transformed Supernova 85.0 d) TK1829 transformed Columbia 125.0 e) TK1829 transformed Supernova 834.0 f) TK1829 transformed Supernova with 1456.0 Norflurazon A comparison of western blot data on various plant extracts allowed a determination of the relative difference in luciferase protein concentration among the various plant lines (Table 2).","Western blot analysis confirmed that Arabidopsis transformed with the TK1829 construct expressed a much greater amount of luciferase expression as compared to plants transformed with a 35S:Luc construct (see Table 2b and d) and as compared to cab2:Luc-transformed Snv (“Supernova”; see Table 2, c and e).","The various plant lines also were examined by imaging analysis using the EGG Berthold Night Owl imaging system (see, also, Example 2C, below).","Plants were sprayed with approximately 1 ml of 5 mM luciferin in a solution of 0.001% TRITON X-100 detergent.","The camera settings were kept consistent for all plant lines.","However, the time duration for taking the images was varied to accommodate the different levels of bioluminescence in the various plant lines.","All data was processed and averaged for photon counts of light/area over the time the image was taken (mm/sec); variation between individual plants was eliminated by analyzing pools of plants.","TABLE 2 Fold increase in luciferase Plant Comparison protein concentration a) Supernova vs. 35S:Luc 5 b) TK1829 transformed Columbia 128 vs. 35S:Luc c) TK1829 transformed Columbia vs. 31 Supernova d) TK1829 transformed Supernova vs. 244 35S:Luc e) TK1829 transformed Supernova vs. 53 Supernova f) TK1829 transformed Supernova with 2 Norflurazon vs. the same line without g) TK1829 transformed Supernova with 450 Norflurazon vs. 35S:Luc In parallel with the levels of luc+ expression as disclosed above, light emission by the TK1829 transformed plants was substantially greater than that of Columbia plants transformed with the 35S:Luc construct (“35S:Luc”) or of Snv plants transformed with a cab2:Luc construct (“Supernova”; see Table 3).","Light emission of Arabidopsis plants transformed with TK1829, as measured by photon counts per area per second, was increased by as much as 5,200 fold over 35S:Luc transformed Columbia plants (see Table 3g).","Plants also were inspected by eye to confirm that they were visibly bioluminescent.","The TK1829 transformed Columbia and TK1829-transformed Supernova plants were visibly bioluminescent (whether grown in the presence or absence of norflurazon), whereas the none of the other plants, including the 35S:Luc transformed Columbia and cab2:Luc transformed Supernova plants, were visibly bioluminescent.","These results demonstrate that transformation of a plant with a construct that can express about 125 pg luc+/μg protein as determined by western blot analysis can effectively generate visibly bioluminescent plants.","TABLE 3 Fold increase in light Plant Comparison (cts/mm2/sec) a) Supernova vs. 35S:Luc 70 b) Tk1829 transformed Columbia vs. 35S:Luc 106 c) Tk1829 transformed Columbia vs. 1.5 Supernova d) Tk1829 transformed Supernova vs. 712 35S:Luc e) TK1829 transformed Supernova vs. 10 Supernova f) TK1829 transformed Supernova with 7 Norflurazon vs. Supernova g) TK1829 transformed Supernova with 5,200 Norflurazon vs. 35S:Luc C. Threshold Photon Emission for Visible Bioluminescence In order to provide a standard for determining the threshold number of visible light photons required for visible bioluminescence, the number of photons of visible light emitted per square millimeter per second was determined using a defined system.","A Night Owl camera with WinLight Program LB981 (EG&G Berthold; Bad Wildbad, Germany) was used, and all images were acquired under the following conditions: Cosmic Suppression, ON; Defect Correction, ON; Star Killer, OFF; Camera Readout, SLOW; Gain, HIGH; Look up Table set to GAMMA.","Measurements were taken in a photography dark room with a plate-to-lens distance of 275 mm (standard setting on Night Owl camera).","Each plant was imaged for a time appropriate to the amount of bioluminescence emitted and values were normalized to photon counts/area/second.","Approximate photons/mm2/sec were calculated using a conversion of discharges per minute divided by counts per minute.","The Night Owl camera system was calibrated using manufacturer-provided standards from the Beckman LS6500 scintillation system.","Two standards were used.","The first standard contained scintillant alone, which represents background levels of discharges per minute (dpm).","The second standard contained an amount of carbon-14 in scintillant that produces 100,600 dpm.","Images were taken using 1.1 pixel binning with background subtraction, and values of counts per second (cps) were acquired.","The cps then was multiplied by 60 to obtain counts per minute (cpm).","Each discharge is representative of the energy released by a disintegrating beta particle in the form of a photon.","“Dpm” divided by “cpm” then provided an approximate efficiency for the camera, which was used for all data values to estimate actual photons emitted.","For example, where the carbon-14 standard=3.6 cps and scintillant alone=2.2 cps, the difference=1.4 cps (actual cps)=84 cpm; and 100,600 dpm divided by 84 cpm=1,198 discharges/count (photons/count), which is used as a conversion factor to calculate the number of photons emitted by plant and detected by the camera Using the above conditions and calculation, various wild type and transformed plants were examined and photons/mm2/sec were determined.","No visibly detectable bioluminescence was observed in the Columbia wt plants, the 35S LUC-transformed plants or Supernova plants, whereas all of the TK1829 transformed plants were visibly bioluminescent.","As shown in Table 4, a threshold value of visible bioluminescence occurs between about 700,000 photons/mm2/sec and 1×106 photons/mm2/sec (compare Tables 4c and 4d; corresponding to 126.4 and 1562.4 pg luc+/μg protein, respectively, as determined by luminometry and spectrophotometry; see Example 2B).","TABLE 4 Photon Emission Plant Comparison (cts/mm2/sec) a) Columbia wild-type 16.8 b) 35S:Luc 10,063.0 c) Supernova 701,669.0 d) Tk1829-19-transformed Columbia 1,061,668.0 e) TK1829-11-transformed Supernova 7,160,446.0 f) TK1829-11-transformed Supernova 52,604,180.0 with Norflurazon These results demonstrate that the amount of light produced by the various transformed plants correlates with the amount of luciferase expressed in the plants, and establish standard conditions defining a threshold level of light emission required for visible bioluminescence.","These results further provide guidelines for generating other constructs, which contain various combinations of regulatory elements, that can be examined for the ability to confer visible bioluminescence on a plant.","For example, by comparing a plant transformed with a recombinant nucleic acid molecule comprising a nucleotide sequence encoding a bioluminescent polypeptide operatively linked to one or more regulatory elements, with plants transformed with a 35S:Luc construct or a cab2:Luc construct, recombinant nucleic acid molecules encompassed within the present invention and useful for generating visibly bioluminescent plant cells or transgenic plants comprising such cells can be obtained.","More directly, by transforming plants with various constructs and determining photon emission under standardized conditions as described above, visibly bioluminescent plants can be obtained.","Furthermore, the results disclosed herein indicate that plant cells other than Arabidopsis plant cells can be genetically modified using, for example, a TK1829 recombinant nucleic acid molecule, thus providing a means to obtain different species, strains and varieties of visibly bioluminescent transgenic plants.","In this respect, genetically modified petunia plants that exhibit exceptional bioluminescence also have been generated using the disclosed compositions and methods.","D. Preparation and Characterization of Bioluminescent Petunia Plants pMT-OM-LUC+ or pACT-OM-LUC+ expression constructs were introduced from MT-OM-LUC+ or pACT-OM-LUC+ into Agrobacterium strain ABI 1 and in planta transformation of Arabidopsis and of petunia plants were performed as described above.","Whole leaves from 10 week old petunia plants were removed and washed in warm soapy water.","Leaves were sterilized in 10% (v/v)chlorine for 15 min and rinsed 4 times with sterile water.","Squares of approximately 1 cm were cut from the leaves under sterile conditions with an scalpel and cultured for one day in pre-culture medium (MS medium supplemented with 1 mg/l of 6-benzylaminopurine (BAP) and 0.1 μg/ml of alpha-naphthalene acetic acid (NAA).","A. tumefaciens strain ABI carrying one of the plasmids was used to inoculate the pieces of leaves.","Bacteria from a single colony were grown at 28° C. overnight in liquid LB medium on a rotary shaker (250 rpm).","The medium was supplemented with 50 μg/ml kanamycin, 100 μg/ml spectinomycin and 34 μg/ml chloramphenicol.","The next day, a 1:10 dilution was made into LB medium (without antibiotics) and sub-cultured cells were grown for 4 to 5 hr.","Cultures were diluted to an OD (660 nm) of 0.05 to 0.07 with pre-culture liquid medium.","Explants were inoculated with a log phase solution of Agrobacterium adjusted to approximately 5×108 cells.","The leaf pieces were allow to soak for 5 min in the bacteria solution, blotted on sterile Whatman filter discs to remove excess of solution, then replaced in the original pre-culture plates and incubated for 2 days at 23° C. under low light.","The leaf squares were transferred into shoot proliferation medium (pre-culture medium containing 75 μg/ml gentamycin and 500 μg/ml carbenicillin).","After 4 weeks, shoots were excised and placed on rooting medium (MS medium supplemented with 60 μg/ml gentamycin and 500 μg/ml carbenicillin).","After approximately 5 weeks, the plants were transferred to soil.","As shown in Table 5, Arabidopsis plants containing the MT-OM-LUC+ construct (designated “31” in Table 5) or the ACT-OM-LUC+ construct (“#27” and “#17”) were visibly biolunminescent (i.e., greater than 750,000 photons of light/mm2/sec (column labeled “photons”).","These results demonstrate that the methods of the invention can be used to make various constructs, which, when introduced into plant cells, result in the generation of visibly bioluminescent plants.","Similarly, introduction of the TK 1829 construct or the ACT-OM-LUC+ construct into petunia plants resulted in the generation of bioluminescent petunia plants.","Although petunia plants containing the Mr-OM-LUC+ construct were not examined for photon counts, the mean RLU/μg was similar to those for the TK 1828 and ACT-OM-LUC+ transformed petunias and the plants were visibly bioluminescent to the eye.","These results demonstrate that the constructs of the invention are useful for generating a variety of visibly bioluminescent plants.","Together, the results of these studies demonstrate the broad utility of the claimed methods for generating visibly bioluminescent plants.","Although the invention has been described with reference to the above examples, it will be understood that modifications and variations are encompassed within the spirit and scope of the invention.","Accordingly, the invention is limited only by the following claims.","TABLE 5 21 day old Weight OD OD conc.","mean pgLuc/ plants ug (750 nm) (750 nm) (ug/ul) RLU 1 RLU 2 RLU/ug ug cts/mm2/sec photons MT Line 31902 31 99.3 0.086 0.086 0.0238844 128.1 132.8 1092.34395 15.24 2408.4 2,885,263.20 Actin Lines Actin 2-7 Col 31902 #27 98.9 0.047 0.046 0.0111278 38.37 39.6 700.68 9.80 911.64 1,092,144.72 Actin 1-1 Col 31902 #17 98.2 0.109 0.109 0.0313123 318.4 331.5 2,075.54 28.81 3342.4 4,004,195.20 Petunia Actin Line T3 102 0.062 0.065 5.0861595 31.46 30.96 1.23 0.02 840 1,006,320.00 TK1829 Line T5 96 0.105 101 84.933336 525.9 520.1 1.23 0.02 3385.7 4,056,068.60 MT Line T3 90 0.106 0.105 3.6981111 22.82 20.23 1.16 0.02 n/a control 27 0.089 0.09 0.0150032 0.028 0.021 0.33 0.00 16.35 19,587.30"]],"string":"[\n [\n \"Bioluminescent plants and methods of making same\",\n \"A genetically modified plant cell containing a heterologous nucleotide sequence encoding a bioluminescent polypeptide, which is expressed in amount sufficient to produce at least about 750,000 photons of visible light/mm2/second, is provided, as is a visibly bioluminescent transgenic plant, which contains such a genetically modified plant cell.\",\n \"Also provided is a recombinant nucleic acid molecule, which contains a plant translational enhancer operatively linked to a nucleotide sequence encoding a bioluminescent polypeptide.\",\n \"In addition, methods of producing a genetically modified plant cell that is visibly bioluminescent introducing a transgene encoding a bioluminescent polypeptide into a plant cell, whereby the bioluminescent polypeptide is expressed at a level that produces at least about 750,000 photons of visible light/mm2/second are provided, as are kits containing such visibly bioluminescent compositions.\"\n ],\n [\n \"1.A genetically modified plant cell, comprising a heterologous nucleotide sequence encoding a bioluminescent polypeptide, wherein the bioluminescent polypeptide can be expressed in amount sufficient to produce at least 750,000 photons of visible light/mm2/second.\",\n \"2.The genetically modified plant cell of claim 1, wherein the bioluminescent polypeptide is luciferase.\",\n \"3.The genetically modified plant cell of claim 1, wherein the bioluminescent polypeptide is a luciferase variant.\",\n \"4.The genetically modified plant cell of claim 3, wherein the luciferase variant is a luc+ luciferase variant.\",\n \"5.The genetically modified plant cell of claim 4, wherein the luc+ luciferase variant is expressed in the plant cell at a level of at least 100 pg/μg protein as determined by western blot analysis.\",\n \"6.The genetically modified plant cell of claim 1, wherein the bioluminescent polypeptide can be expressed in amount sufficient to produce at least one million photons of visible light/mm2/second.\",\n \"7.The genetically modified plant cell of claim 1, which has a reduced level of chlorophyll as compared to corresponding wild type plant cell.\",\n \"8.A transgenic plant, comprising the genetically modified plant cell of claim 1 or claim 7.9.A plant cell or tissue obtained from the transgenic plant of claim 8.10.A cutting of the transgenic plant of claim 8.11.A seed produced by the transgenic plant of claim 8.12.A cDNA or genomic DNA library prepared from the transgenic plant of claim 8, or from a plant cell or plant tissue obtained from said transgenic plant.\",\n \"13.The transgenic plant of claim 8, wherein the plant is a monocot.\",\n \"14.The transgenic plant of claim 8, wherein the plant is a dicot.\",\n \"15.The transgenic plant of claim 14, wherein the plant is an angiosperm.\",\n \"16.The transgenic plant of claim 15, wherein the angiosperm is a cereal plant, a leguminous plant, an oilseed plant, or a hardwood tree.\",\n \"17.The transgenic plant of claim 8, wherein the plant is an ornamental plant.\",\n \"18.The transgenic plant of claim 17, wherein the ornamental plant is a petunia.\",\n \"19.The transgenic plant of claim 17, wherein the ornamental plant is a carnation.\",\n \"20.The genetically modified plant of claim 1, wherein the heterologous nucleotide sequence encoding a bioluminescent polypeptide comprises nucleotides 8366 to 11,113 of SEQ ID NO:2.21.The genetically modified plant of claim 1, wherein the heterologous nucleotide sequence encoding a bioluminescent polypeptide comprises nucleotides 8340 to 12,098 of SEQ ID NO:4 or nucleotides 6 to 3570 of SEQ ID NO:5.22.A recombinant nucleic acid molecule, comprising, in operative linkage, a plant translational enhancer and a nucleotide sequence encoding a bioluminescent polypeptide.\",\n \"23.The recombinant nucleic acid molecule of claim 22, wherein the plant translational enhancer is a plant potyvirus translational enhancer.\",\n \"24.The recombinant nucleic acid molecule of claim 23, wherein the plant potyvirus is a tobacco etch virus (TEV) or an omega enhancer.\",\n \"25.The recombinant nucleic acid molecule of claim 22, wherein the bioluminescent polypeptide is a luciferase polypeptide or variant thereof.\",\n \"26.The recombinant nucleic acid molecule of claim 22, further comprising an operatively linked transcriptional regulatory element, which comprises a promoter.\",\n \"27.The recombinant nucleic acid molecule of claim 26, wherein the transcriptional regulatory element is a cauliflower mosaic virus (CaMV) 35S regulatory element, a metallothionein regulatory element, or an actin 2 regulatory element.\",\n \"28.The recombinant nucleic acid molecule of claim 22, comprising, in operative linkage, a CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and a CaMV 35S terminator.\",\n \"29.The recombinant nucleic acid molecule of claim 28, wherein the CaMV 35S enhancer is a dual CaMV 35S enhancer.\",\n \"30.The recombinant nucleic acid molecule of claim 28, wherein the luciferase polypeptide or variant thereof is a luc+ polypeptide.\",\n \"31.The recombinant nucleic acid molecule of claim 22, comprising, in operative linkage, a metallothionein gene transcriptional regulatory element, an omega translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and an RbcS E9 polyA sequence.\",\n \"32.The recombinant nucleic acid molecule of claim 22, comprising, in operative linkage, an actin 2 gene transcriptional regulatory element, an omega translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and an RbcS E9 polyA sequence.\",\n \"33.The recombinant nucleic acid molecule of claim 22, comprising nucleotides 8366 to 11,113 of SEQ ID NO:2.34.The recombinant nucleic acid molecule of claim 22, comprising nucleotides 8340 to 12,098 of SEQ ID NO:4 or nucleotides 6 to 3570 of SEQ ID NO:5.35.A vector, comprising the recombinant nucleic acid molecule of claim 22.36.The vector of claim 32, wherein the vector is an expression vector.\",\n \"37.The vector of claim 35, wherein the recombinant nucleic acid molecule comprises, in operative linkage, a CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and a CaMV 35S terminator.\",\n \"38.The vector of claim 35, wherein the recombinant nucleic acid molecule comprises, in operative linkage, a metallothionein gene transcriptional regulatory element, an omega translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and an RbcS E9 polyA sequence.\",\n \"39.The vector of claim 35, wherein the recombinant nucleic acid molecule comprises, in operative linkage, an actin 2 gene transcriptional regulatory element, an omega translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and an RbcS E9 polyA sequence.\",\n \"40.The vector of claim 35, wherein the recombinant nucleic acid molecule comprises nucleotides 8366 to 11,113 of SEQ ID NO:2.41.The vector of claim 35, wherein the recombinant nucleic acid molecule comprises nucleotides 8340 to 12, 098 of SEQ ID NO:4 or nucleotides 6 to 3570 of SEQ ID NO:5.42.The vector of claim 35, which is selected from SEQ ID NO:1 and SEQ ID NO:2.43.The vector of claim 35, which is selected from SEQ ID NO:4 and SEQ ID NO:5.44.A cell containing the recombinant nucleic acid molecule of claim 22.45.A cell containing the vector of claim 35.46.A method of producing a genetically modified plant cell that is visibly bioluminescent, the method comprising introducing a transgene comprising a nucleotide sequence encoding a bioluminescent polypeptide into a plant cell, whereby the bioluminescent polypeptide is expressed at a level that produces at least 750,000 photons of visible light/mm2/second, thereby producing a genetically modified plant cell that visibly bioluminesce.\",\n \"47.The method of claim 45, wherein the transgene further comprises a plant translational enhancer operatively linked to the nucleotide sequence encoding a bioluminescent polypeptide.\",\n \"48.The method of claim 46, wherein the plant translational enhancer is a plant potyvirus translational enhancer selected from a tobacco etch virus (TEV) translational enhancer and a tobacco mosaic virus translational enhancer.\",\n \"49.The method of claim 46, wherein the plant translational enhancer is an omega translational enhancer having a nucleotide sequence set forth as nucleotides 1169 to 1235 of SEQ ID NO:5.50.The method of claim 46, wherein the bioluminescent polypeptide is a luciferase polypeptide or variant thereof.\",\n \"51.The method of claim 47, wherein the transgene further comprises a transcriptional regulatory element operatively linked to the plant translational enhancer and nucleotide sequence encoding a bioluminescent polypeptide.\",\n \"52.The method of claim 51, wherein the transcriptional regulatory element is a cauliflower mosaic virus (CaMV) 35S regulatory element, a metallothionein gene regulatory element, or an actin gene regulatory element.\",\n \"53.The method of claim 46, wherein the transgene comprises, in operative linkage, a dual CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and a CaMV 35S terminator.\",\n \"54.The method of claim 53, wherein the luciferase polypeptide or variant thereof is a luc+ polypeptide.\",\n \"55.The method of claim 46, wherein the transgene comprises, in -operative linkage, a metallothionein gene transcriptional regulatory element, which comprises a promoter and, optionally, an enhancer; an omega translational enhancer; a nucleotide sequence encoding a luciferase polypeptide or variant thereof; and an RbcS E9 polyA sequence.\",\n \"56.The method of claim 55, wherein the luciferase polypeptide or variant thereof is a luc+ polypeptide.\",\n \"57.The method of claim 46, wherein the transgene comprises, in operative linkage, an actin 2 gene transcriptional regulatory element, which comprises a promoter and, optionally, an enhancer; an omega translational enhancer; a nucleotide sequence encoding a luciferase polypeptide or variant thereof; and an RbcS E9 polyA sequence.\",\n \"58.The method of claim 57, wherein the luciferase polypeptide or variant thereof is a luc+ polypeptide.\",\n \"59.The method of claim 46, wherein the transgene comprises nucleotides 8366 to 11,113 of SEQ ID NO:2, nucleotides 8340 to 12,098 of SEQ ID NO:4 or nucleotides 6 to 3570 of SEQ ID NO:5.60.The method of claim 46, wherein the transgene is stably maintained in the plant cell genome.\",\n \"61.The method of claim 60, wherein the transgene is integrated into the plant cell genome.\",\n \"62.The method of claim 46, further comprising contacting the plant cell, or the genetically modified plant cell derived therefrom, with an agent that inhibits carotenoid synthesis, thereby reducing the amount of chlorophyll in the plant cell.\",\n \"63.The method of claim 62, wherein the agent is norfluorazon.\",\n \"64.A genetically modified plant cell produced by the method of claim 46 or claim 62.65.A transgenic plant comprising the genetically modified plant cell of claim 64.66.A genetically modified plant cell, comprising a heterologous nucleotide, which comprises, in operative linkage, a CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and a CaMV 35S terminator wherein the luciferase polypeptide or variant thereof can be expressed in amount sufficient to produce at least 750,000 photons of visible light/mm2/second.\",\n \"67.The genetically modified plant cell of claim 66, wherein the heterologous nucleotide sequence encoding a bioluminescent polypeptide comprises nucleotides 8366 to 11,113 of SEQ ID NO:2.68.A transgenic plant, comprising the genetically modified plant cell of claim 66 or claim 67.69.A plant cell or tissue obtained from the transgenic plant of claim 68.70.A cutting of the transgenic plant of claim 68.71.A seed produced by the transgenic plant of claim 68.72.A cDNA or genomic DNA library prepared from the transgenic plant of claim 68, or from a plant cell or plant tissue obtained from said transgenic plant.\",\n \"73.A genetically modified plant cell, comprising a heterologous nucleotide sequence, which comprises, in operative linkage, a metallothionein gene transcriptional regulatory element, an omega translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and an RbcS E9 polyA sequence., wherein the luciferase polypeptide or variant thereof can be expressed in amount sufficient to produce at least 750,000 photons of visible light/mm2/second.\",\n \"74.The genetically modified plant cell of claim 73, wherein the heterologous nucleotide sequence comprises nucleotides 8340 to 12,098 of SEQ ID NO:4 75.A transgenic plant, comprising the genetically modified plant cell of claim 73 or claim 74.76.A plant cell or tissue obtained from the transgenic plant of claim 75.77.A cutting of the transgenic plant of claim 75.78.A seed produced by the transgenic plant of claim 75.79.A genetically modified plant cell, comprising a heterologous nucleotide sequence, which comprises, in operative linkage, an actin 2 gene transcriptional regulatory element, an omega translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and an RbcS E9 polyA sequence, wherein the luciferase polypeptide or variant thereof can be expressed in amount sufficient to produce at least 750,000 photons of visible light/mm2/second.\",\n \"80.The genetically modified plant cell of claim 79, wherein the heterologous nucleotide sequence comprises nucleotides 6 to 3570 of SEQ ID NO:5.81.A transgenic plant, comprising the genetically modified plant cell of claim 79 or claim 80.82.A plant cell or tissue obtained from the transgenic plant of claim 81.83.A cutting of the transgenic plant of claim 81.84.A seed produced by the transgenic plant of claim 81.85.A kit, comprising the genetically modified plant cell of claim 1 or claim 7, or a derivative of the genetically modified plant cell.\",\n \"86.A kit comprising a derivative of the genetically modified plant cell, wherein the derivative of the genetically modified plant cell is a transgenic plant comprising the genetically modified plant cell of claim 1 or claim 7, or a cell, tissue or organ of said transgenic plant.\",\n \"87.The kit of claim 86, wherein the organ of said transgenic plant is a flower or a bract.\",\n \"88.The kit of claim 85, further comprising an amount of luciferin sufficient for generating visible bioluminescence of the genetically modified plant cell or the derivative of the genetically modified plant cell.\",\n \"89.The kit of claim 85, further comprising an amount of a carotenoid inhibitor sufficient for reducing or inhibiting chlorophyll production in the genetically modified plant cell or the derivative of the genetically modified plant cell.\",\n \"90.A kit, comprising a cutting or a seed of the transgenic plant of claim 8.91.The kit of claim 90, further comprising reagents for growing a transgenic plant from the cutting or seed.\",\n \"92.The kit of claim 90, further comprising an amount of a carotenoid inhibitor sufficient for reducing or inhibiting chlorophyll production in a transgenic plant grown from the cutting or seed.\",\n \"93.The kit of claim 90, further comprising an amount of luciferin sufficient for generating visible bioluminescence of a transgenic plant grown from the cutting or seed.\",\n \"94.A kit, comprising the genetically modified plant cell of claim 66, 67, 73, 74, 79, or 80, or a derivative of the genetically modified plant cell.\",\n \"95.A kit comprising a derivative of a genetically modified plant cell, wherein the derivative of the genetically modified plant cell is a transgenic plant comprising the genetically modified plant cell of claim 66, 67, 73, 74, 79 or 80, or a cell, tissue or organ of said transgenic plant.\",\n \"96.The kit of claim 95, wherein the organ of said transgenic plant is a flower or a bract.\",\n \"97.The kit of claim 94, further comprising an amount of luciferin sufficient for generating visible bioluminescence of the genetically modified plant cell or the derivative of the genetically modified plant cell.\",\n \"98.The kit of claim 94, further comprising an amount of a carotenoid inhibitor sufficient for reducing or inhibiting chlorophyll production in the genetically modified plant cell or the derivative of the genetically modified plant cell.\",\n \"99.A kit, comprising a cutting of the transgenic plant of claim 68, or a seed produced by said transgenic plant.\",\n \"100.The kit of claim 99, further comprising reagents for growing a transgenic plant from the cutting or seed.\",\n \"101.The kit of claim 99, further comprising an amount of a carotenoid inhibitor sufficient for reducing or inhibiting chlorophyll production in a transgenic plant grown from the cutting or seed.\",\n \"102.The kit of claim 99, further comprising an amount of luciferin sufficient for generating visible bioluminescence of a transgenic plant grown from the cutting or seed.\",\n \"103.The kit of claim 95, further comprising an amount of luciferin sufficient for generating visible bioluminescence of the genetically modified plant cell or the derivative of the genetically modified plant cell.\",\n \"104.The kit of claim 95, further comprising an amount of a carotenoid inhibitor sufficient for reducing or inhibiting chlorophyll production in the genetically modified plant cell or the derivative of the genetically modified plant cell.\",\n \"105.A kit comprising a cutting of the transgenic plant of claim 75, or a seed produced by said transgenic plant.\",\n \"106.The kit of claim 105, further comprising reagents for growing a transgenic plant from the cutting or seed.\",\n \"107.The kit of claim 105, further comprising an amount of a carotenoid inhibitor sufficient for reducing or inhibiting chlorophyll production in a transgenic plant grown from the cutting or seed.\",\n \"108.The kit of claim 105, further comprising an amount of luciferin sufficient for generating visible bioluminescence of a transgenic plant grown from the cutting or seed.\",\n \"109.A kit comprising a cutting of the transgenic plant of claim 81, or a seed produced by said transgenic plant.\",\n \"110.The kit of claim 109, further comprising reagents for growing a transgenic plant from the cutting or seed.\",\n \"111.The kit of claim 110 further comprising an amount of a carotenoid inhibitor sufficient for reducing or inhibiting chlorophyll production in a transgenic plant grown from the cutting or seed.\",\n \"112.The kit of claim 109, further comprising an amount of luciferin sufficient for generating visible bioluminescence of a transgenic plant grown from the cutting or seed.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>The present invention relates to a genetically modified plant cell that exhibits bioluminescence that is visible to the naked eye.\",\n \"Such a genetically modified plant cell contains a heterologous nucleotide sequence that encodes a bioluminescent polypeptide, which, upon expression, can result in the production of at least about 750,000 photons of visible light/mm 2 /second by the plant cell.\",\n \"The bioluminescent polypeptide can be any polypeptide that is bioluminescent, or can act in concert with a second molecule such that visible light is produced.\",\n \"In one embodiment, the genetically modified plant cell contains a heterologous nucleotide sequence encoding luciferase or a luciferase variant, for example, luc + .\",\n \"Upon contact of a genetically modified plant expressing luciferase or a variant thereof with luciferin, a sufficient amount of visible light is produced such that the light is visible to the naked eye.\",\n \"Such a genetically modified plant cell generally expresses at least about 100 pg luc + /μg protein as determined by western blot analysis such that, upon contact of the plant cell with luciferin, the cell is visibly bioluminescent.\",\n \"In another embodiment, the genetically modified plant cell, or the plant cell from which the genetically modified plant cell is derived, is contacted with an agent that reduces or inhibits pigment formation in the plant, for example, carotenoid synthesis, thereby reducing or inhibiting chlorophyll synthesis.\",\n \"Since chlorophyll in a plant cell can absorb photons generated by the luciferase in the genetically modified plant cell, contact of the genetically modified plant cell, or a transgenic plant derived therefrom, with an amount of a carotenoid inhibitor sufficient to reduce or inhibit chlorophyll synthesis provides a means to enhance photon emission from the plant cell and, therefore, enhance the visible bioluminescence.\",\n \"In still another embodiment, a visibly bioluminescent transgenic plant is generated from the genetically modified plant cell containing a heterologous nucleotide sequence encoding a bioluminescent polypeptide.\",\n \"As such, the invention relates to such a transgenic plant, as well as to a plant cell or tissue obtained from the transgenic plant, a seed produced by the transgenic plant, and a cDNA or genomic DNA library prepared from the transgenic plant or from a plant cell or plant tissue obtained from the transgenic plant.\",\n \"The transgenic plant can be a monocot, or can be a dicot, for example, an angiosperm such as a cereal plant, a leguminous plant, an oilseed plant, a hardwood tree, or an ornamental plant such as a petunia, an orchid, a carnation or the like.\",\n \"The present invention also relates to a recombinant nucleic acid molecule, which comprises a translational enhancer, which enhances translation in a plant cell, operatively linked to a nucleotide sequence encoding a bioluminescent polypeptide.\",\n \"The translational enhancer can be any translational enhancer, for example, a plant potyvirus translational enhancer such as tobacco etch virus (TEV) translational enhancer, a tobacco mosaic virus omega translational enhancer, or a translational enhancer comprising a Kozak sequence.\",\n \"The encoded bioluminescent polypeptide can be a luciferase polypeptide or variant thereof, for example, the luc + luciferase variant.\",\n \"A recombinant nucleic acid molecule of the invention also can contain a transcriptional regulatory element, i.e., promoter, enhancer, etc., that enhances transcription in a plant cell, for example, a plant virus enhancer such as a cauliflower mosaic virus (CaMV) 35S enhancer, or a regulatory element from any other organism, for example, an actin 2 regulatory element or a metallothionein regulatory element, which comprises a promoter and, optionally, an enhancer, wherein the transcriptional regulatory element is operatively linked to the plant translational enhancer and nucleotide sequence encoding the bioluminescent polypeptide.\",\n \"In one embodiment, a recombinant nucleic acid molecule contains, in operative linkage, a CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and a CaMV 35S terminator.\",\n \"In another embodiment, the CaMV 35S enhancer is a dual CaMV 35S enhancer, and the luciferase polypeptide or variant thereof is the luc + luciferase variant.\",\n \"Such a recombinant nucleic acid molecule is exemplified herein by the nucleotide sequence set forth as nucleotides 8366 to 11,113 of SEQ ID NO:2.In still other embodiments, the transcriptional regulatory element in a recombinant nucleic molecule of the invention is a metallothionein regulatory element or an actin 2 regulatory element, and the translational enhancer is an omega enhancer, for example, in a recombinant nucleic acid molecule having a sequence set forth as nucleotides 8340 to 12,098 of SEQ ID NO:4 or as nucleotides 6 to 3570 of SEQ ID NO:5.The present invention further relates to a vector containing a recombinant nucleic acid molecule comprising a translational enhancer active in a plant operatively linked to a nucleotide sequence encoding a bioluminescent polypeptide.\",\n \"Preferably, the vector is an expression vector.\",\n \"In one embodiment, the recombinant nucleic acid molecule contained in the vector comprises, in operative linkage, a CAMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and a CaMV 35S terminator.\",\n \"In another embodiment, the recombinant nucleic acid molecule contained in the vector comprises, in operative linkage, a dual CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luc + luciferase variant, and a CaMV 35S terminator.\",\n \"In still another embodiment, the recombinant nucleic acid molecule contained in the vector comprises the nucleotide sequence set forth as nucleotides 8366 to 11,113 of SEQ ID NO:2.In yet another embodiment, the transcriptional regulatory element in the recombinant nucleic acid molecule comprises an actin 2 promoter or a metallothionein promoter, and the translation enhancer is an omega enhancer, for example, in the recombinant nucleic acid molecules set forth as nucleotides 8340 to 12,098 of SEQ D) NO:4 or as nucleotides 6 to 3570 of SEQ ID NO:5.Vectors of the invention are exemplified herein by those as set forth in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:4 and SEQ ID NO:5.The present invention also provides a cell containing a recombinant nucleic acid molecule of the invention, or containing a vector of the invention.\",\n \"The present invention further relates to a method of producing a genetically modified plant cell that is visibly bioluminescent.\",\n \"A method of the invention can be performed, for example, by introducing a transgene comprising a nucleotide sequence encoding a bioluminescent polypeptide into a plant cell, whereby the bioluminescent polypeptide is expressed at a level that produces at least about 750,000 photons of visible light/mm 2 /second.\",\n \"The transgene useful in a method of making a genetically modified plant cell that is visibly bioluminescent generally includes a translational enhancer that is active in a plant, for example, a plant translational enhancer such as a plant potyvirus translational enhancer (e.g., a TEV translational enhancer), a tobacco mosaic virus (TMV) translational enhancer such as the omega enhancer, operatively linked to the nucleotide sequence encoding a bioluminescent polypeptide, for example, a luciferase polypeptide or variant thereof.\",\n \"The transgene also can contain a plant virus enhancer, for example, a CaMV 35S enhancer, which is operatively linked to the plant translational enhancer and nucleotide sequence encoding a bioluminescent polypeptide.\",\n \"In one embodiment, a transgene useful in making a visibly luminescent genetically modified plant cell contains, in operative linkage, a dual CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and a CaMV 35S terminator.\",\n \"In another embodiment, the nucleotide sequence encodes a luc + luciferase variant polypeptide.\",\n \"Such transgenes are exemplified by the nucleotide sequences set forth as nucleotides 8366 to 11,113 of SEQ ID NO:2, nucleotides 8340 to 12,098 of SEQ ID NO:4, and nucleotides 6 to 3570 of SEQ ID NO:5.A method of the invention can further include contacting the plant cell, or the genetically modified plant cell derived therefrom, with an agent that inhibits the synthesis of a pigment such as carotenoids, which can absorb light generated by the luciferase.\",\n \"For example, the genetically modified plant cell can be contacted with an agent such as norfluorazon, which reduces or inhibits chlorophyll production in the plants, thus allowing increased emission of photons from the plant cell, or from a trrnsgenic plant derived from the plant cell.\",\n \"A visibly bioluminescent genetically modified plant cell made according to a method of the invention generally contains the transgene stably maintained in the plant cell genome.\",\n \"Preferably, the transgene is integrated into the plant cell genome.\",\n \"As such, the present invention also relates to a genetically modified plant cell produced by a method of the invention, as well as to a transgenic plant containing or generated from such a genetically modified plant cell.\",\n \"The present invention also relates to kit, which contains a genetically modified plant cell of the invention, or a derivative of the genetically modified plant cell, for example, a transgenic plant derived from the genetically modified plant cell, or a cell, a tissue, or an organ of such a transgenic plant.\",\n \"As such, a kit of the invention can contain one or more flowers, bracts, leaves or other tissues or organs, which, upon contact with luciferin, are visibly luminescent.\",\n \"Accordingly, a kit of the invention can further include an amount of luciferin sufficient for generating visible bioluminescence of the genetically modified plant cell or the derivative of the genetically modified plant cell, and also can include a plurality of such amounts of luciferin, thus allowing the generation of visible bioluminesce a number of times, as desired.\",\n \"In addition, a kit of the invention can include an amount of a inhibitor that reduces or inhibits pigment synthesis in the genetically modified plant cell or the derivative of the genetically modified plant cell.\",\n \"For example, the kit can contain a carotenoid synthesis inhibitor such as norfluorazon, which reduces or inhibits chlorophyll production.\",\n \"In another embodiment, a kit of the invention contains one or more cuttings, seeds, or other portion or derivative of a visibly bioluminescent plant of the invention such that a visibly luminescent transgenic plant can be grown therefrom.\",\n \"As such, the kit also can include reagents for growing a transgenic plant from the cutting or seed, including, for example, a suitable plant food or other nutrient source required for growth of the particular plant.\",\n \"In addition, the kit can include an amount of a carotenoid inhibitor sufficient for reducing or inhibiting chlorophyll production in a trrnsgenic plant grown from the cutting or seed; and/or can include an amount of luciferin sufficient for generating visible bioluminescence of a transgenic plant grown from the cutting or seed.\"\n ],\n [\n \"BACKGROUND OF THE INVENTION FIELD OF THE INVENTION The present invention relates generally to plant biology and more specifically to plants that are genetically modified to exhibit bioluminescence, and to compositions and methods for making such genetically modified bioluminescent plants.\",\n \"BACKGROUND INFORMATION Plants and plant products provide the primary sustenance, either directly or indirectly, for all animal life, including humans.\",\n \"For the majority of the world's human population and for many animals, plants and plant products provide the sole source of nutrition.\",\n \"As the world population increases, the best hope to prevent widespread famine is to increase the quantity and improve the quality of food crops.\",\n \"Throughout history, a continual effort has been made to increase the yield and nutritious value of food crops.\",\n \"For centuries, plants having desirable characteristics such as greater resistance-to drought conditions or increased size of fruit were crossbred and progeny plants exhibiting the desired characteristics were selected and used to produce seed or cuttings for propagation.\",\n \"Using such classical genetic methods, plants having, for example, greater disease resistance, increased yield, and better flavor have been obtained.\",\n \"More recently, the methods of genetic engineering have been applied to plant biology, thus expediting the generation of plants having desired characteristics.\",\n \"For example, genetically engineered tomato plants have been generated such that the tomatoes do not continue to ripen after they have been picked from the plant.\",\n \"As a result, there is no need to pick unripe tomatoes and hope that they attain an optimum level of ripeness at the time the consumer is ready to eat them.\",\n \"Instead, the tomatoes can be picked at a time when they are near their optimal ripeness, then shipped to consumers for consumption.\",\n \"Genetically engineered plants that are less susceptible to freezing or to damage by biological pests also have been generated.\",\n \"The availability of such plants has been a boon to the agricultural industry, and provides a measure of certainty that sufficient quantities of food will remain available in the future.\",\n \"Methods of genetic engineering also have impacted the ornamental plant industry.\",\n \"In addition to protecting ornamental plants from environmental stresses or biological pests, genetic engineering has been used to produce ornamental plants having new and interesting characteristics.\",\n \"For example, genetic engineering has been used to generate roses that are blue in color.\",\n \"The ability to generate ornamental plants having new and unusual characteristics provides a means to expand the ornamental plant market and, therefore, the economy in general.\",\n \"Thus, a need exists to identify interesting and unusual traits that can be expressed by a plant and can create a new market in the ornamental plant industry.\",\n \"The present invention satisfies this need and provides additional advantages.\",\n \"SUMMARY OF THE INVENTION The present invention relates to a genetically modified plant cell that exhibits bioluminescence that is visible to the naked eye.\",\n \"Such a genetically modified plant cell contains a heterologous nucleotide sequence that encodes a bioluminescent polypeptide, which, upon expression, can result in the production of at least about 750,000 photons of visible light/mm2/second by the plant cell.\",\n \"The bioluminescent polypeptide can be any polypeptide that is bioluminescent, or can act in concert with a second molecule such that visible light is produced.\",\n \"In one embodiment, the genetically modified plant cell contains a heterologous nucleotide sequence encoding luciferase or a luciferase variant, for example, luc+.\",\n \"Upon contact of a genetically modified plant expressing luciferase or a variant thereof with luciferin, a sufficient amount of visible light is produced such that the light is visible to the naked eye.\",\n \"Such a genetically modified plant cell generally expresses at least about 100 pg luc+/μg protein as determined by western blot analysis such that, upon contact of the plant cell with luciferin, the cell is visibly bioluminescent.\",\n \"In another embodiment, the genetically modified plant cell, or the plant cell from which the genetically modified plant cell is derived, is contacted with an agent that reduces or inhibits pigment formation in the plant, for example, carotenoid synthesis, thereby reducing or inhibiting chlorophyll synthesis.\",\n \"Since chlorophyll in a plant cell can absorb photons generated by the luciferase in the genetically modified plant cell, contact of the genetically modified plant cell, or a transgenic plant derived therefrom, with an amount of a carotenoid inhibitor sufficient to reduce or inhibit chlorophyll synthesis provides a means to enhance photon emission from the plant cell and, therefore, enhance the visible bioluminescence.\",\n \"In still another embodiment, a visibly bioluminescent transgenic plant is generated from the genetically modified plant cell containing a heterologous nucleotide sequence encoding a bioluminescent polypeptide.\",\n \"As such, the invention relates to such a transgenic plant, as well as to a plant cell or tissue obtained from the transgenic plant, a seed produced by the transgenic plant, and a cDNA or genomic DNA library prepared from the transgenic plant or from a plant cell or plant tissue obtained from the transgenic plant.\",\n \"The transgenic plant can be a monocot, or can be a dicot, for example, an angiosperm such as a cereal plant, a leguminous plant, an oilseed plant, a hardwood tree, or an ornamental plant such as a petunia, an orchid, a carnation or the like.\",\n \"The present invention also relates to a recombinant nucleic acid molecule, which comprises a translational enhancer, which enhances translation in a plant cell, operatively linked to a nucleotide sequence encoding a bioluminescent polypeptide.\",\n \"The translational enhancer can be any translational enhancer, for example, a plant potyvirus translational enhancer such as tobacco etch virus (TEV) translational enhancer, a tobacco mosaic virus omega translational enhancer, or a translational enhancer comprising a Kozak sequence.\",\n \"The encoded bioluminescent polypeptide can be a luciferase polypeptide or variant thereof, for example, the luc+ luciferase variant.\",\n \"A recombinant nucleic acid molecule of the invention also can contain a transcriptional regulatory element, i.e., promoter, enhancer, etc., that enhances transcription in a plant cell, for example, a plant virus enhancer such as a cauliflower mosaic virus (CaMV) 35S enhancer, or a regulatory element from any other organism, for example, an actin 2 regulatory element or a metallothionein regulatory element, which comprises a promoter and, optionally, an enhancer, wherein the transcriptional regulatory element is operatively linked to the plant translational enhancer and nucleotide sequence encoding the bioluminescent polypeptide.\",\n \"In one embodiment, a recombinant nucleic acid molecule contains, in operative linkage, a CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and a CaMV 35S terminator.\",\n \"In another embodiment, the CaMV 35S enhancer is a dual CaMV 35S enhancer, and the luciferase polypeptide or variant thereof is the luc+ luciferase variant.\",\n \"Such a recombinant nucleic acid molecule is exemplified herein by the nucleotide sequence set forth as nucleotides 8366 to 11,113 of SEQ ID NO:2.In still other embodiments, the transcriptional regulatory element in a recombinant nucleic molecule of the invention is a metallothionein regulatory element or an actin 2 regulatory element, and the translational enhancer is an omega enhancer, for example, in a recombinant nucleic acid molecule having a sequence set forth as nucleotides 8340 to 12,098 of SEQ ID NO:4 or as nucleotides 6 to 3570 of SEQ ID NO:5.The present invention further relates to a vector containing a recombinant nucleic acid molecule comprising a translational enhancer active in a plant operatively linked to a nucleotide sequence encoding a bioluminescent polypeptide.\",\n \"Preferably, the vector is an expression vector.\",\n \"In one embodiment, the recombinant nucleic acid molecule contained in the vector comprises, in operative linkage, a CAMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and a CaMV 35S terminator.\",\n \"In another embodiment, the recombinant nucleic acid molecule contained in the vector comprises, in operative linkage, a dual CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luc+ luciferase variant, and a CaMV 35S terminator.\",\n \"In still another embodiment, the recombinant nucleic acid molecule contained in the vector comprises the nucleotide sequence set forth as nucleotides 8366 to 11,113 of SEQ ID NO:2.In yet another embodiment, the transcriptional regulatory element in the recombinant nucleic acid molecule comprises an actin 2 promoter or a metallothionein promoter, and the translation enhancer is an omega enhancer, for example, in the recombinant nucleic acid molecules set forth as nucleotides 8340 to 12,098 of SEQ D) NO:4 or as nucleotides 6 to 3570 of SEQ ID NO:5.Vectors of the invention are exemplified herein by those as set forth in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:4 and SEQ ID NO:5.The present invention also provides a cell containing a recombinant nucleic acid molecule of the invention, or containing a vector of the invention.\",\n \"The present invention further relates to a method of producing a genetically modified plant cell that is visibly bioluminescent.\",\n \"A method of the invention can be performed, for example, by introducing a transgene comprising a nucleotide sequence encoding a bioluminescent polypeptide into a plant cell, whereby the bioluminescent polypeptide is expressed at a level that produces at least about 750,000 photons of visible light/mm2/second.\",\n \"The transgene useful in a method of making a genetically modified plant cell that is visibly bioluminescent generally includes a translational enhancer that is active in a plant, for example, a plant translational enhancer such as a plant potyvirus translational enhancer (e.g., a TEV translational enhancer), a tobacco mosaic virus (TMV) translational enhancer such as the omega enhancer, operatively linked to the nucleotide sequence encoding a bioluminescent polypeptide, for example, a luciferase polypeptide or variant thereof.\",\n \"The transgene also can contain a plant virus enhancer, for example, a CaMV 35S enhancer, which is operatively linked to the plant translational enhancer and nucleotide sequence encoding a bioluminescent polypeptide.\",\n \"In one embodiment, a transgene useful in making a visibly luminescent genetically modified plant cell contains, in operative linkage, a dual CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and a CaMV 35S terminator.\",\n \"In another embodiment, the nucleotide sequence encodes a luc+ luciferase variant polypeptide.\",\n \"Such transgenes are exemplified by the nucleotide sequences set forth as nucleotides 8366 to 11,113 of SEQ ID NO:2, nucleotides 8340 to 12,098 of SEQ ID NO:4, and nucleotides 6 to 3570 of SEQ ID NO:5.A method of the invention can further include contacting the plant cell, or the genetically modified plant cell derived therefrom, with an agent that inhibits the synthesis of a pigment such as carotenoids, which can absorb light generated by the luciferase.\",\n \"For example, the genetically modified plant cell can be contacted with an agent such as norfluorazon, which reduces or inhibits chlorophyll production in the plants, thus allowing increased emission of photons from the plant cell, or from a trrnsgenic plant derived from the plant cell.\",\n \"A visibly bioluminescent genetically modified plant cell made according to a method of the invention generally contains the transgene stably maintained in the plant cell genome.\",\n \"Preferably, the transgene is integrated into the plant cell genome.\",\n \"As such, the present invention also relates to a genetically modified plant cell produced by a method of the invention, as well as to a transgenic plant containing or generated from such a genetically modified plant cell.\",\n \"The present invention also relates to kit, which contains a genetically modified plant cell of the invention, or a derivative of the genetically modified plant cell, for example, a transgenic plant derived from the genetically modified plant cell, or a cell, a tissue, or an organ of such a transgenic plant.\",\n \"As such, a kit of the invention can contain one or more flowers, bracts, leaves or other tissues or organs, which, upon contact with luciferin, are visibly luminescent.\",\n \"Accordingly, a kit of the invention can further include an amount of luciferin sufficient for generating visible bioluminescence of the genetically modified plant cell or the derivative of the genetically modified plant cell, and also can include a plurality of such amounts of luciferin, thus allowing the generation of visible bioluminesce a number of times, as desired.\",\n \"In addition, a kit of the invention can include an amount of a inhibitor that reduces or inhibits pigment synthesis in the genetically modified plant cell or the derivative of the genetically modified plant cell.\",\n \"For example, the kit can contain a carotenoid synthesis inhibitor such as norfluorazon, which reduces or inhibits chlorophyll production.\",\n \"In another embodiment, a kit of the invention contains one or more cuttings, seeds, or other portion or derivative of a visibly bioluminescent plant of the invention such that a visibly luminescent transgenic plant can be grown therefrom.\",\n \"As such, the kit also can include reagents for growing a transgenic plant from the cutting or seed, including, for example, a suitable plant food or other nutrient source required for growth of the particular plant.\",\n \"In addition, the kit can include an amount of a carotenoid inhibitor sufficient for reducing or inhibiting chlorophyll production in a trrnsgenic plant grown from the cutting or seed; and/or can include an amount of luciferin sufficient for generating visible bioluminescence of a transgenic plant grown from the cutting or seed.\",\n \"BRIEF DESCRIPTION OF THE FIGURES FIG.\",\n \"1 shows a map of the pTK1829 plasmid.\",\n \"Restriction endonuclease sites are indicated, as are the first nucleotide positions of the sites, except that the first and last nucleotide positions of the multiple cloning site are indicated.\",\n \"“luc+” indicates region encoding variant luciferase polypeptide; “Amp” indicates ampicillin-resistance gene (see, also, SEQ ID NO:1).\",\n \"FIG.\",\n \"2 shows a map of the pPZPTK1829 binary vector plasmid.\",\n \"Restriction endonuclease sites and nucleotide positions are indicated as in FIG.\",\n \"1.“luc+” indicates region encoding variant luciferase polypeptide; “Spec” and “Gent-R” indicate the genes conferring resistance to spectinomycin and gentamycin, respectively.\",\n \"“Ori” indicates the plasmid origin of replication.\",\n \"“LB” and “RB” indicate left border and right border, respectively, of the binary vector that allow transfer of the insert into the plant (see, also, SEQ ID NO:2).\",\n \"FIG.\",\n \"3 shows a map of the MT-OM-LUC+ binary vector plasmid.\",\n \"Restriction endonuclease sites and nucleotide positions are indicated as in FIG.\",\n \"1.“luc+”, “Spec”, “Gent-R”, “Ori”, and “LB” and “RB” are as in FIG.\",\n \"2.“MT” indicates the metallothionein transcriptional regulatory element; “Ome” indicates the omega translational enhancer; and “E9 3′” indicates the RbcS E9 polyA region (see, also, SEQ ID NO:4).\",\n \"FIG.\",\n \"4 shows a map of the ACT-OM-LUC+ binary vector plasmid.\",\n \"Restriction endonuclease sites and nucleotide positions are indicated as in FIG.\",\n \"1.“luc+”, “Spec”, “Gent-R”, “ori”, “LB” and “RB” are as in FIG.\",\n \"2.“Ome” and “E9 3′” are as in FIG.\",\n \"3; “ACT” indicates the actin 2 transcriptional regulatory element (see, also, SEQ ID NO:5).\",\n \"DETAILED DESCRIPTION OF THE INVENTION The present invention provides genetically modified plant cells that are visibly bioluminescent due to expression of heterologous polypeptide, and visibly bioluminescent transgenic plants comprising or generated from such genetically modified plant cells.\",\n \"As disclosed herein, the transgenic plants of the invention exhibit bioluminescence that is visible to the human eye.\",\n \"Prior to the present disclosure, plant cells could be genetically modified to express heterologous bioluminescent polypeptides such as luciferase, but light gathering or amplifying instrumentation was required to detect the bioluminescence; one could not simply look at plant comprising such cells directly and detect the bioluminescence.\",\n \"It is noted that there is a report on the world wide web, at the URL “hybridorchids.com”, of a bioluminescent orchid.\",\n \"However, no published information for making such orchids is available and the information on the website has not been updated since 2000.The present invention provides compositions and methods that obviate the previous limitations that prevented the generation of visibly detectable plants and plant cells.\",\n \"As disclosed herein, the visibly bioluminescent genetically modified plant cells and transgenic plants of the invention are useful as research tools, and have great ornamental value.\",\n \"A genetically modified plant cell or transgenic plant of the invention is visibly bioluminescent due to expression in the plant cell, or in plant cells of the transgenic plant, of a bioluminescent polypeptide.\",\n \"As used herein, the term “bioluminescent polypeptide” refers to a polypeptide that can be expressed in a living organism and exhibits or effects the emission of visible light.\",\n \"A bioluminescent polypeptide can inherently emit visible light such as fluorescence, or can be contacted with a physical or chemical agent such that a reaction occurs and visible light is produced.\",\n \"For example, a genetically modified plant cell can contain a heterologous nucleotide sequence encoding luciferase such that, upon expression of the luciferase and contact of the plant cell with luciferin, a chemiluminescent reaction occurs and visible light is emitted.\",\n \"As used herein, the term “visible light” refers to electromagnetic radiation associated with the visible portion of the electromagnetic spectrum, generally electromagnetic radiation having a wavelength between about 100 to 1000 nanometers (nm), and particularly between about 350 to 750 nm.\",\n \"Luciferase polypeptides and luciferase variants are examples of bioluminescent polypeptides that are particularly useful for practicing the methods and producing the compositions of the invention.\",\n \"As used herein, the term “luciferase variant” refers to a modified luciferase polypeptide that contains one or more amino acid additions, deletions or substitutions as compared to a corresponding wild type luciferase polypeptide, and that, when contacted with an appropriate substrate such as luciferin, undergoes a reaction that results in-the production of visible light.\",\n \"Luciferases include bacterial luciferases such as Vibrio harveyi or V. fisheri luciferase, marine ostracod crustacean luciferases such a Vargula hilgendorii luciferase, dinoflagellate luciferases such as Gonyaulax ploydra luciferase, coelenterate luciferases such as Renilla renifornis (sea pansy) luciferase, or Coleoptera (beetle) luciferases such as an Elateridae (click beetle) family luciferase, a Phengodidae (glow worm) family luciferase, or a Lampyridae (firefly) luciferase, for example, a Photintus carolinus or P. pyralis luciferase, and the like, including modified forms thereof (see, for example, U.S. Pat.\",\n \"No.\",\n \"4,968,613; U.S. Pat.\",\n \"No.\",\n \"5,221,623; U.S. Pat.\",\n \"No.\",\n \"5,229,285; U.S. Pat.\",\n \"No.\",\n \"5,330,906; U.S. Pat.\",\n \"No.\",\n \"5,604,123; U.S. Pat.\",\n \"No.\",\n \"5,618,722; U.S. Pat.\",\n \"No 5,674,713; U.S. Pat.\",\n \"No.\",\n \"5,814,465; and U.S. Pat.\",\n \"No.\",\n \"5,843,746, each of which is incorporated herein by reference).\",\n \"A luciferase variant is exemplified by the firefly luciferase variant, luc+ (U.S. Pat.\",\n \"No.\",\n \"5,670,356; Sherf and Wood, Promega Notes 49:14, 1994, each of which is incorporated herein by reference), which lacks a peroxisome localizing domain, can be particularly useful for preparing a genetically modified plant cell or visibly bioluminescent transgenic plant of the invention.\",\n \"Polynucleotide sequences encoding luciferases and mutant luciferases are well known (see above references) and vectors containing such polynucleotides are commercially available (Promega; Madison Wis.).\",\n \"As used herein, the term “visibly bioluminescent” refers to the emission of a sufficient amount of visible light such that it can be seen by a human eye.\",\n \"As such, the use of light gathering, light amplifying, light enhancing or light accumulating instrumentation is not required to detect the bioluminescence, although such instrumentation can be used, for example, to quantitate the amount of visible light emitted.\",\n \"It should be recognized that a visibly bioluminescent plant cell may not be visible to the unaided eye due to its small size, but can be seen using a magnifying glass, microscope, or the like.\",\n \"Such a plant cell is considered to be visibly bioluminescent for purposes of the present invention because light gathering, accumulating or amplifying instrumentation such as a luminometer, photomultiplier, image enhancing video system, spectrophotometer, film emulsion, digitizing equipment, or the like is not required.\",\n \"The light produced by a visibly bioluminescent transgenic plant can be seen by a person or other mammal looking at the plant, particularly when the plant is in a dark environment, for example, in a darkened room or outside at night.\",\n \"In addition, a bioluminescent plant of the invention can be photographed using a camera, including any generally used camera such as a 35 mm single lens reflex camera, a digital camera, a Polaroid® camera, and the like, such that a picture of the glowing plant can be obtained.\",\n \"As disclosed herein, a genetically modified plant cell, or a plant, plant tissue or plant organ comprising the plant cell that emits at least about 750,00 photons of visible light/mm2second, generally at least about 850,000 photons of visible light/mm2/second, and particularly at least about 1×106 (one million) photons of visible light/mm2/second is visibly bioluminescent (see Example 2C, Table 4).\",\n \"As disclosed herein, contact of a genetically modified plant cell of the invention, or of a plant cell from which the genetically modified plant cell is derived, or of a transgenic plant of the invention with an agent that reduces or inhibits the synthesis of one or more pigments in the plant or plant cell.\",\n \"By reducing the amount of pigmentation of a plant, or a portion of the plant such as the leaves or flowers, absorption of bioluminescence by the pigment is reduced, thereby enhancing the emission of photons from the plant and, therefore, the visible bioluminescence.\",\n \"For example, contact of the genetically modified plant cell or a transgenic plant of the invention with an inhibitor of carotenoid synthesis can reduce or inhibit chlorophyll synthesis in the plant, thus reducing absorption of photons by chlorophyll in the plant and allowing greater emission of the photons from the plant.\",\n \"Inhibitors of carotenoid synthesis in plants are well known and include herbicides such as norfluorazone, which is commercially available as ZORIAL® herbicide (Syngenta Crop Production; Greensboro N.C.).\",\n \"In another embodiment, a visibly bioluminescent transgenic plant is generated from the genetically modified plant cell containing a heterologous nucleotide sequence encoding a bioluminescent polypeptide.\",\n \"As such, the invention relates to such a transgenic plant, as well as to a plant cell or tissue obtained from the transgenic plant, a seed produced by the transgenic plant, and a cDNA or genomic DNA library prepared from the trrnsgenic plant or from a plant cell or plant tissue obtained from the transgenic plant.\",\n \"The transgenic plant can be a monocot, or can be a dicot, for example, an angiosperm such as a cereal plant, a leguminous plant, an oilseed plant, a hardwood tree, or an ornamental plant such as a petunia, a carnation or the like.\",\n \"In one embodiment, plants useful for purposes of the present invention include orchids.\",\n \"In another embodiment, plants useful for purposes of the present invention exclude orchids.\",\n \"The present invention provides a recombinant nucleic acid molecule, which includes a heterologous nucleotide sequence encoding a bioluminescent polypeptide operatively linked to one or more transcriptional or translational regulatory elements, or a combination thereof.\",\n \"As used herein, the term “heterologous” is used in a comparative sense with respect to a nucleotide sequence encoding a bioluminescent polypeptide to indicate either that the nucleotide sequence is not an endogenous nucleotide sequence in a plant cell into which it is to be introduced, or that the nucleotide sequence is part of a construct such that it is in a form other than it normally would be found in a plant cell in nature.\",\n \"For example, a heterologous nucleotide sequence can encode a firefly luciferase, which is not normally expressed in a plant cell and, therefore, is heterologous with respect to the plant cell.\",\n \"The regulatory element or combination of regulatory elements is selected such that the bioluminescent polypeptide can be expressed in a plant cell at a level sufficient for the plant cell, or a plant comprising the cell, to visibly bioluminesce.\",\n \"The term “regulatory element” is used broadly herein to refer to a nucleotide sequence that, when operatively linked to a second, expressible nucleotide effects transcription or translation of the nucleotide sequence such that a ribonucleic acid (RNA) molecule or polypeptide, respectively, can be generated, or encodes a domain that confers a desirable characteristic on a polypeptide containing the domain.\",\n \"For purposes of the present invention, a regulatory element generally increases the amount of transcription or translation of an operatively linked nucleotide sequence encoding the bioluminescent polypeptide, or provides a means to localize a bioluminescent polypeptide to a particular location in a cell, particularly a plant cell.\",\n \"Transcriptional and translational regulatory elements are well known and include promoters, enhancers, 3′-untranslated or 5′-untranslated sequences of transcribed sequence, for example, a poly-A signal sequence or other transcription termination signal, or other protein or RNA stabilizing elements, or other gene expression control elements known to regulate gene expression or the amount of expression of a gene product.\",\n \"A regulatory element useful for constructing a recombinant nucleic acid molecule of the invention can be isolated from a naturally occurring genomic DNA sequence or can be synthetic, for example, a synthetic promoter.\",\n \"Transcriptional regulatory elements can be constitutively expressed regulatory elements, which maintain gene expression at a relatively constant level of activity, or can be inducible regulatory elements.\",\n \"Constitutively expressed regulatory elements can be expressed in any cell type, for example an actin promoter such as an actin 2 promoter, or an elongation factor (EF) promoter such as an EF1α promoter; or can be a tissue specific regulatory element, which is expressed only in one or a few specific cell types, a phase specific regulatory element, which is expressed only during particular developmental or growth stages of a plant cell, or the like.\",\n \"A regulatory element such as a tissue specific or phase specific regulatory element or an inducible regulatory element useful in constructing a recombinant nucleic acid molecule of the invention can be a regulatory element that generally, in nature, is found in a plant genome.\",\n \"However, the regulatory element also can be from an organism other than a plant, including, for example, from a plant virus, an animal virus, or a cell from an animal or other multicellular organism.\",\n \"As used herein, the term “tissue specific or phase specific regulatory element” means a nucleotide sequence that effects transcription in only one or a few cell types, or only during one or a few stages of the life cycle of a plant, for example, only for a period of time during a particular stage of growth, development or differentiation.\",\n \"The terms “tissue specific” and “phase specific” are used together herein in referring to such a regulatory element because a single regulatory element can have characteristics of both types of regulatory elements.\",\n \"For example, a regulatory element active only during a particular stage of plant development also can be expressed only in one or a few types of cells in the plant during the particular stage of development.\",\n \"A tissue specific or phase specific regulatory element can be useful for producing a genetically modified plant that is bioluminescent only in particular organs or tissues, or only at a particular developmental stage.\",\n \"Numerous tissue specific or phase specific regulatory elements have been described and are known and available to those in the art.\",\n \"Such tissue or phase specific regulatory elements include, for example, the AGL8/FRUITFULL regulatory element, which is activated upon floral induction (Hempel et al., Development 124:3845-3853, 1997, which is incorporated herein by reference); root specific regulatory elements such as the regulatory elements from the RCP1 gene and the LRP1 gene (Tsugeki and Fedoroff, Proc.\",\n \"Natl.\",\n \"Acad.\",\n \"Sci., USA 96:12941-12946, 1999; Smith and Fedoroff, Plant Cell 7:735-745, 1995, each of which is incorporated herein by reference); flower specific regulatory elements such as the regulatory elements from the LEAFY gene and the APETELA1 gene (Blazquez et al., Development 124:3835-3844, 1997, which is incorporated herein by reference; Hempel et al., supra, 1997); seed specific regulatory elements such as the regulatory element from the oleosin gene (Plant et al., Plant Mol.\",\n \"Biol.\",\n \"25:193-205, 1994, which is incorporated herein by reference), and dehiscence zone specific regulatory element.\",\n \"Additional tissue specific or phase specific regulatory elements include the Zn13 promoter, which is a pollen specific promoter (Hamilton et al., Plant Mol.\",\n \"Biol.\",\n \"18:211-218, 1992, which is incorporated herein by reference); the UNUSUAL FLORAL ORGANS (UFO) promoter, which is active in apical shoot meristem; the promoter active in shoot meristems (Atanassova et al., Plant J.\",\n \"2:291, 1992, which is incorporated herein by reference), the cdc2a promoter and cyc07 promoter (see, for example, Ito et al., Plant Mol.\",\n \"Biol.\",\n \"24:863, 1994; Martinez et al., Proc.\",\n \"Natl.\",\n \"Acad.\",\n \"Sci., USA 89:7360, 1992; Medford et al., Plant Cell 3:359, 1991; Terada et al., Plant J.\",\n \"3:241, 1993; Wissenbach et al., Plant J.\",\n \"4:411, 1993, each of which is incorporated herein by reference); the promoter of the APETELA3 gene, which is active in floral meristems (Jack et al., Cell 76:703, 1994, which is incorporated herein by reference; Hempel et al., supra, 1997); a promoter of an agamous-like (AGL) family member, for example, AGL8, which is active in shoot meristem upon the transition to flowering (Hempel et al., supra, 1997); floral abscission zone promoters; L1-specific promoters; and the like.\",\n \"Such tissue specific or phase specific regulatory elements can be used to construct a recombinant nucleic acid useful for producing, for example, a transgenic plant that is visibly bioluminescent in one or a few selected tissues or organs such as in flowers or in leaves, or that are expressed only during a particular stage of development such as during flowering.\",\n \"Inducible regulatory elements also are useful for constructing a recombinant nucleic acid molecule of the invention.\",\n \"As used herein, the term “inducible regulatory element” means a regulatory element that, when exposed to an inducing agent, effects an increased level of transcription of an operatively linked nucleotide sequence encoding the bioluminescent polypeptide as compared to the level of transcription, if any, in the absence of an inducing agent.\",\n \"Inducible regulatory elements can be those that have no basal or constitutive activity and only effect transcription upon exposure to an inducing agent, or those that effect a basal or constitutive level of transcription, which is increased upon exposure to an inducing agent.\",\n \"Inducible regulatory elements that effect a basal or constitutive level of expression generally are particularly useful where the induced level of transcription is substantially greater than the basal or constitutive level of expression, for example, at least about two-fold greater, or at least about five-fold greater.\",\n \"Particularly useful inducible regulatory elements do not have a basal or constitutive activity, or increase the level of transcription at least about ten-fold greater than a basal or constitutive level of transcription associated with the regulatory element.\",\n \"The term “inducing agent” is used to refer to a chemical, biological or physical agent that effects transcription from an inducible regulatory element.\",\n \"In response to exposure to an inducing agent, transcription from the inducible regulatory element generally is initiated de novo or is increased above a basal or constitutive level of expression.\",\n \"Such induction can be identified using the methods disclosed herein, including detecting an increased level of mRNA encoding the bioluminescent polypeptide.\",\n \"The use of an inducible regulatory element in a recombinant nucleic acid molecule of the invention provides a means to express the bioluminescent polypeptide only at a desired time, thus preventing extraneous transcriptional or translational activity in the plant cell.\",\n \"An inducing agent useful in a method of the invention is selected based on the particular inducible regulatory element.\",\n \"For example, the inducible regulatory element can be a metallothionein (MT) regulatory element such as an MT2B regulatory element, a copper inducible regulatory element, or a tetracycline inducible regulatory element, the transcription from which can be effected in response to various metal ions, to copper or to tetracycline, respectively (Furst et al., Cell 55:705-717, 1988; Mett et al., Proc.\",\n \"Natl.\",\n \"Acad.\",\n \"Sci., USA 90:4567-4571, 1993; Gatz et al., Plant J.\",\n \"2:397-404, 1992; Roder et al., Mol.\",\n \"Gen. Genet.\",\n \"243:32-38, 1994, each of which is incorporated herein by reference).\",\n \"The inducible regulatory element also can be an ecdysone regulatory element or a glucocorticoid regulatory element, the transcription from which can be effected in response to ecdysone or other steroid (Christopherson et al., Proc.\",\n \"Natl.\",\n \"Acad.\",\n \"Sci., USA 89:6314-6318, 1992; Schena et al., Proc.\",\n \"Natl.\",\n \"Acad.\",\n \"Sci., USA 88:10421-10425, 1991, each of which is incorporated herein by reference).\",\n \"In addition, the regulatory element can be a cold responsive regulatory element or a heat shock regulatory element, the transcription of which can be effected in response to exposure to cold or heat, respectively (Takahashi et al., Plant Physiol.\",\n \"99:383-390, 1992, which is incorporated herein by reference).\",\n \"Additional regulatory elements useful in the methods or compositions of the invention include, for example, the spinach nitrite reductase gene regulatory element (Back et al., Plant Mol.\",\n \"Biol.\",\n \"17:9, 1991, which is incorporated herein by reference); a light inducible regulatory element (Feinbaum et al., Mol.\",\n \"Gen. Genet.\",\n \"226:449, 1991; Lam and Chua, Science 248:471, 1990, each of which is incorporated herein by reference), a plant hormone inducible regulatory element (Yamaguchi-Shinozaki et al., Plant Mol.\",\n \"Biol.\",\n \"15:905 (1990); Kares et al., Plant Mol.\",\n \"Biol.\",\n \"15:225 (1990), each of which is incorporated herein by reference), and the like.\",\n \"A circadian rhythm regulatory element such as a cab2 promoter (see, for example, Millar et al., Plant Cell 4:1075-1087, 1992, which is incorporated herein by reference) can also be considered an example of an inducible regulatory element, since expression from such a promoter appears to occur in response to a factor associated with a time of day.\",\n \"A regulatory element operatively linked to a nucleotide sequence encoding a bioluminescent polypeptide also can comprise an enhancer, including a transcriptional enhancer, translational enhancer, or both.\",\n \"A transcriptional enhancer useful for purposes of the present invention can be any enhancer generally associated with a plant gene or an enhancer from another organism that has enhancer activity in a plant cell, for example, a plant virus enhancer, or an enhancer from a gene expressed in an invertebrate cell, or a vertebrate cell such as a mammalian cell, or an enhancer of a virus that infects such cells.\",\n \"A cauliflower mosaic virus (CaMV) 35S enhancer is an example of a plant virus enhancer that is useful for increasing the transcriptional activity of a nucleotide sequence to which it is operatively linked.\",\n \"Thus, in one embodiment, a recombinant nucleic acid molecule of the invention-contains a CaMV 35S enhancer operatively linked to a nucleotide sequence encoding the bioluminescent polypeptide.\",\n \"Preferably, the CaMV 35S enhancer is a dual enhancer, comprising two copies of the CaMV 35S enhancer nucleotide sequence.\",\n \"An actin 2 regulatory element provides another example of a transcriptional regulatory element, comprising a promoter and an enhancer, that is useful in constructing a composition of the invention or practicing a method of the invention (see nucleotides 6 to 1145 of SEQ ID NO:5).\",\n \"A metallothionein regulatory element provides another example of a transcriptional regulatory element useful for purposes of the present invention, and provides the additional advantage that it includes an inducible enhancer (see nucleotides 11,000 to 12,098 of SEQ ID NO:4).\",\n \"A translational enhancer, which enhances translation of a polypeptide in a plant cell (also referred to generally herein as a plant translational enhancer), useful in a recombinant nucleic acid molecule of the invention similarly can be any translational enhancer that generally is associated with a plant gene or with a gene of another organism.\",\n \"Examples of a plant translational enhancer include a plant potyvirus translational enhancer such as the tobacco etch virus (TEV) translational enhancer, and a tobacco mosaic virus translational enhancer such as the omega enhancer (see, for example, nucleotides 1169 to 1235 of SEQ ID NO:5; see, also, Carrington and Freed, J. Virol.\",\n \"64:1590, 1990; Gallie et al., Plant Cell 1:301-311, 1989, each of which is incorporated herein by reference), as well as the nucleotide sequence beginning about 101 nucleotide upstream of the potato virus S (PSV) coat protein gene (Turner and Foster, Arch.\",\n \"Virol.\",\n \"142:167-175, 1997, which is incorporated herein by reference).\",\n \"A recombinant nucleic acid molecule of the invention can also contain regulatory elements required for termination of transcription, for example, a polyadenylation signal.\",\n \"A CaMV 35S terminator is an example of a transcription termination signal useful in constructing a recombinant nucleic acid molecule.\",\n \"A recombinant nucleic acid molecule of the invention also can contain regulatory elements required for initiation or termination of translation or both, including, for example, an internal ribosome entry site (IRES), a Kozak sequence, a nucleotide sequence encoding an initiator methionine residue, and a nucleotide sequence encoding a STOP codon, which terminates translation.\",\n \"The regulatory element also can be a nucleotide sequence encoding a leader sequence, signal peptide, cell compartmentalization domain, or the like, and can be included in the construct, for example, where it is desired to localize the bioluminescent polypeptide to a particular compartment such as the cytosol, nucleus, a chloroplast, or another subcellular organelle in a plant cell.\",\n \"Such intracellular localization can be particularly useful where the genetically modified cell, or plant containing the cell, is to be used as a research tool, for example, a particular intracellular compartment of a plant cell.\",\n \"A translation initiation or termination element or other regulatory element can be contained in the nucleotide sequence encoding the bioluminescent polypeptide can be a regulatory element that is normally associated with the nucleotide sequence in nature and is a part of the nucleotide sequence as a consequence of its isolation from a natural source, or can be heterologous to the nucleotide sequence and operatively linked thereto.\",\n \"Additional regulatory elements such as an intron sequence, for example, from Adh1 or bronzel, or a viral leader sequence, for example, from TMV, MCMV or AIVIV, also enhance expression of a nucleotide sequence encoding a bioluminescent polypeptide and, therefore, can be a component of a recombinant nucleic acid molecule of the invention.\",\n \"As disclosed herein, combinations of regulatory elements can be selected such that a recombinant nucleic acid molecule comprising a nucleotide sequence encoding a bioluminescent polypeptide can be constructed, and wherein the recombinant nucleic acid molecule allows for expression of the bioluminescent polypeptide in a plant cell such that the plant cell is visibly bioluminescent.\",\n \"A recombinant nucleic acid molecule of the invention is exemplified by, in operative linkage, a CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and a CaMV 35S terminator.\",\n \"In one embodiment, the CaMV 35S enhancer comprises a dual CaMV 35S enhancer, and the luciferase polypeptide or variant thereof is a luc+luciferase variant.\",\n \"In another embodiment, a recombinant nucleic acid molecule of the invention is exemplified by the nucleotide sequence set forth as about nucleotides 8366 to 11,113 of SEQ ID NO:2.A recombinant nucleic acid molecule of the invention also is exemplified by the sequence set forth as nucleotides 8340 to 12,098 of SEQ ID NO:4, which includes, in operative linkage, a metallothionein gene (MT2B) regulatory element comprising an enhancer and promoter (nucleotides 11,000 to 12,098 of SEQ ID NO:4), an omega translational enhancer (nucleotides 10,675 to 10,741 of SEQ ID NO:4), a nucleotide sequence encoding a luciferase polyp eptide or variant thereof such as luc+ (nucleotides 9015 to 10,667 or SEQ ID NO:4) and an RbcS E9 polyA region (nucleotides 8340 to 8981 of SEQ ID NO:4).\",\n \"In addition, a recombinant nucleic acid molecule of the invention also is exemplified by the sequence set forth as nucleotides 6 to 3570 of SEQ ID NO:5, which includes, in operative linkage, an actin 2 regulatory element comprising an enhancer and promoter (nucleotides 6 to 1145 of SEQ ID NO:5), an omega translational enhancer (nucleotides 1169 to 1235 of SEQ ID NO:5), a nucleotide sequence encoding a luciferase polypeptide or variant thereof such as luc+ (nucleotides 1243 to 2895 of SEQ ID NO:5), and an RbcS E9 polyA region (nucleotides 2929 to 3570 of SEQ ID NO:5).\",\n \"A recombinant nucleic acid molecule of the invention comprises a nucleotide sequence encoding a bioluminescent polypeptide operatively linked to one or more regulatory elements, for example, a translational enhancer.\",\n \"As used herein, the term “operatively linked,” when used in reference to a regulatory element and a second nucleotide sequence, which can be another regulatory element or a coding or other sequence, means that the regulatory element is positioned with respect to the second nucleotide sequence such that the regulatory element effects its function with respect to the second nucleotide sequence in substantially the same manner as it does when the regulatory element is present in its natural position in a genome.\",\n \"Transcriptional promoters, for example, generally act in a position and orientation dependent manner, and usually are positioned at or within about five nucleotides to about fifty nucleotides 5′ (upstream) of the start site of transcription of a gene in nature.\",\n \"In comparison, enhancers can act in a relatively position or orientation independent manner, and can be positioned several hundred or thousand nucleotides upstream or downstream from a transcription start site, or in an intron within the coding region of a gene, yet still be operatively linked to the coding region so as to enhance transcription.\",\n \"The relative positions and orientations of various regulatory elements, including the positioning of a transcribed regulatory sequence such as an IRES or a translated regulatory element such as a cell compartmentalization domain in an appropriate reading frame, are well known and methods for operatively linking such elements are routine in the art (see, for example, Sambrook et al., “Molecular Cloning: A laboratory manual” (Cold Spring Harbor Laboratory Press 1989); Ausubel et al., “Current Protocols in Molecular Biology” (John Wiley and Sons, Baltimore Md.\",\n \"1987, and supplements through 1995), each of which is incorporated herein by reference).\",\n \"As used herein, the term “nucleic acid molecule” or “polynucleotide” or “nucleotide sequence” means a sequence of two or more deoxyribonucleotides or ribonucleotides that are linked together by a phosphodiester bond.\",\n \"As such, the terms include RNA and DNA, which can be a gene or a portion thereof, a cDNA, a synthetic polydeoxyribonucleic acid sequence, or the like, and can be single stranded or double stranded, as well as a DNA/RNA hybrid.\",\n \"Furthermore, the terms are used herein to include naturally occurring nucleic acid molecules, which can be isolated from a cell, as well as synthetic molecules, which can be prepared, for example, by methods of chemical synthesis or by enzymatic methods such as by the polymerase chain reaction (PCR).\",\n \"The term “recombinant” is used herein to refer to a nucleic acid molecule that is produced by linking together at least two nucleotide sequences that are not generally linked together in nature.\",\n \"As such, a recombinant nucleic acid molecule encompassed within the present invention is distinguishable from a nucleic acid molecule that may be produced, for example, during meiosis.\",\n \"In general, the nucleotides comprising a recombinant nucleic acid molecule, polynucleotide or nucleotide sequence are naturally occurring deoxyribonucleotides, such as adenine, cytosine, guanine or thymine linked to 2′-deoxyribose, or ribonucleotides such as adenine, cytosine, guanine or uracil linked to ribose.\",\n \"However, a nucleic acid molecule or nucleotide sequence also can contain nucleotide analogs, including non-naturally occurring synthetic nucleotides or modified naturally occurring nucleotides.\",\n \"Such nucleotide analogs are well known in the art and commercially available, as are polynucleotides containing such nucleotide analogs (Lin et al., Nucl.\",\n \"Acids Res.\",\n \"22:5220-5234, 1994; Jellinek et al., Biochemistry 34:11363-11372, 1995; Pagratis et al., Nature Biotechnol.\",\n \"15:68-73, 1997, each of which is incorporated herein by reference).\",\n \"Similarly, the covalent bond linking the nucleotides of a nucleotide sequence generally is a phosphodiester bond.\",\n \"However, the covalent bond also can be any of numerous other bonds, including a thiodiester bond, a phosphorothioate bond, a peptide-like bond or any other bond known to those in the art as useful for linking nucleotides to produce synthetic polynucleotides (see, for example, Tam et al., Nucl.\",\n \"Acids Res.\",\n \"22:977-986, 1994; Ecker and Crooke, BioTechnology 13:351360, 1995, each of which is incorporated herein by reference).\",\n \"The incorporation of non-naturally occurring nucleotide analogs or bonds linking the nucleotides or analogs can be particularly useful where the nucleic acid molecule is to be exposed to an environment that can contain a nucleolytic activity, including, for example, a plant tissue culture medium or in a plant cell since the modified molecules can be less susceptible to degradation.\",\n \"A nucleotide sequence containing naturally occurring nucleotides and phosphodiester bonds, can be chemically synthesized or can be produced using recombinant DNA methods, using an appropriate polynucleotide as a template.\",\n \"In comparison, a nucleotide sequence containing nucleotide analogs or covalent bonds other than phosphodiester bonds generally are chemically synthesized, although an enzyme such as T7 polymerase can incorporate certain types of nucleotide analogs into a polynucleotide and, therefore, can be used to produce such a polynucleotide recombinantly from an appropriate template (Jellinek et al., supra, 1995).\",\n \"A recombinant nucleic acid molecule of the invention can be introduced into a cell as a naked DNA molecule, can be incorporated in a matrix such as a liposome or a particle such as a viral particle, or can be incorporated into a vector.\",\n \"Incorporation of the polynucleotide into a vector can facilitate manipulation of the polynucleotide, or introduction of the polynucleotide into a plant cell.\",\n \"Accordingly, the present invention also provides a vector containing a recombinant nucleic acid comprising a nucleotide sequence encoding a bioluminescent polypeptide operatively linked to one or more regulatory elements.\",\n \"A vector can be derived from a plasmid or a viral vector such as a T-DNA vector (Horsch et al., Science 227:1229-1231 (1985), which is incorporated herein by reference).\",\n \"If desired, the vector can comprise components of a plant transposable element, for example, a Ds transposon (Bancroft and Dean, Genetics 134:1221-1229, 1993, which is incorporated herein by reference) or an Spm transposon (Aarts et al., Mol.\",\n \"Gen. Genet.\",\n \"247:555-564, 1995, which is incorporated herein by reference).\",\n \"In addition to containing a recombinant nucleic acid molecule of the invention, the vector also can contain various nucleotide sequences that facilitate, for example, rescue of the vector from a transformed plant cell; passage of the vector in a host cell, which can be a plant, animal, bacterial, or insect host cell; or expression of an encoding nucleotide sequence in the vector, including all or a portion of a rescued coding region.\",\n \"As such, a vector can contain any of a number of additional transcription and translation elements, including constitutive and inducible promoters, enhancers, and the like (see, for example, Bitter et al., Meth.\",\n \"Enzymol.\",\n \"153:516-544, 1987).\",\n \"For example, a vector can contain elements useful for passage, growth or expression in a bacterial system, including a bacterial origin of replication; a promoter, which can be an inducible promoter; and the like.\",\n \"A vector also can contain one or more restriction endonuclease recognition and cleavage sites, including, for example, a polylinker sequence, to facilitate insertion or removal of a recombinant nucleic acid molecule.\",\n \"In addition to a nucleotide sequence encoding a bioluminescent polypeptide, a recombinant nucleic acid molecule of the invention, or a vector containing such a recombinant nucleic acid molecule, can contain one or more other expressible nucleotide sequences encoding an RNA or a polypeptide of interest.\",\n \"For example, the additional nucleotide sequence can encode an antisense nucleic acid molecule; an enzyme such as β-galactosidase, β-glucuronidase, luciferase, alkane phosphatase, glutathione S-transferase, chloramphenicol acetyltransferase, guanine xanthine phosphoribosyltransferase, and neomycin phosphotransferase; a viral polypeptide or a peptide portion thereof; or a growth factor or a hormone, which can be a plant growth factor or hormone.\",\n \"Expression of such a nucleotide sequence can provide a means for selecting for a cell containing the construct, for example, by conferring a desirable phenotype to a plant cell containing the nucleotide sequence.\",\n \"For example, the additional nucleotide sequence can be, or encode, a selectable marker, which, when present or expressed in a plant cell, provides a means to identify the plant cell containing the marker.\",\n \"A selectable marker provides a means for screening a population of plants, or plant cells, to identify those having the marker and, therefore, a recombinant nucleic acid molecule comprising a nucleotide sequence encoding a bioluminescent polypeptide.\",\n \"A selectable marker generally confers a selective advantage to the plant cell, or a plant containing the cell, for example, the ability to grow in the presence of a negative selective agent such as an antibiotic or herbicide.\",\n \"A selective advantage also can be due, for example, to an enhanced or novel capacity to utilize an added compound as a nutrient, growth factor or energy source.\",\n \"A selective advantage can be conferred by a single polynucleotide, or its expression product, or by a combination of polynucleotides whose expression in a plant cell gives the cell a positive selective advantage, a negative selective advantage, or both.\",\n \"It should be recognized that expression of a bioluminescent polypeptide also provides a means to a select plant cell containing the encoding nucleotide sequence.\",\n \"However, the use of an additional selectable marker, which, for example, allows a plant cell to survive under otherwise toxic conditions, provides a means to enrich for transformed plant cells containing the recombinant nucleic acid molecule.\",\n \"Examples of selectable markers include those described above, as well as those that confer antimetabolite resistance, for example, dihydrofolate reductase, which confers resistance to methotrexate (Reiss, Plant Physiol.\",\n \"(Life Sci.\",\n \"Adv.)\",\n \"13:143-149, 1994); neomycin phosphotransferase, which confers resistance to the aminoglycosides neomycin, kanamycin and paromycin (Herrera-Estrella, EMBO J.\",\n \"2:987-995, 1983) and hygro, which confers resistance to hygromycin (Marsh, Gene 32:481-485, 1984), trpB, which allows cells to utilize indole in place of tryptophan; hisD, which allows cells to utilize histinol in place of histidine (Hartman, Proc.\",\n \"Natl.\",\n \"Acad.\",\n \"Sci., USA 85:8047, 1988); mannose-6-phosphate isomerase which allows cells to utilize mannose (WO 94/20627); ornithine decarboxylase, which confers resistance to the ornithine decarboxylase inhibitor, 2-(difluoromethyl)-DL-ornithine (DFMO; McConlogue, 1987, In: Current Communications in Molecular Biology, Cold Spring Harbor Laboratory ed.\",\n \"); and deaminase from Aspergillus terreus, which confers resistance to Blasticidin S (Tamura, Biosci.\",\n \"Biotechnol.\",\n \"Biochem.\",\n \"59:2336-2338, 1995).\",\n \"Additional selectable markers include those that confer herbicide resistance, for example, phosphinothricin acetyltransferase gene, which confers resistance to phosphinothricin (White et al., Nucl.\",\n \"Acids Res.\",\n \"18:1062, 1990; Spencer et al., Theor.\",\n \"Appl.\",\n \"Genet.\",\n \"79:625-631, 1990), a mutant EPSPV-synthase, which confers glyphosate resistance (Hinchee et al., BioTechnology 91:915-922, 1998), a mutant acetolactate synthase, which confers imidazolione or sulfonylurea resistance (Lee et al., EMBO J.\",\n \"7:1241-1248, 1988), a mutant psbA, which confers resistance to atrazine (Smeda et al., Plant Physiol.\",\n \"103:911-917, 1993), or a mutant protoporphyrinogen oxidase (see U.S. Pat.\",\n \"No.\",\n \"5,767,373), or other markers conferring resistance to an herbicide such as glufosinate.\",\n \"In addition, markers that facilitate identification of a plant cell containing the polynucleotide encoding the marker include, for example, luciferase (Giacomin, Plant Sci.\",\n \"116:59-72, 1996; Scikantha, J. Bacteriol.\",\n \"178:121, 1996), green fluorescent protein (Gerdes, FEBS Lett.\",\n \"389:4447, 1996) or fl-glucuronidase (Jefferson, EMBO J.\",\n \"6:3901-3907, 1997), and numerous others as disclosed herein or otherwise known in the art.\",\n \"Such markers also can be used as reporter molecules.\",\n \"As disclosed herein, a vector of the invention contains a recombinant nucleic acid molecule comprising one or more regulatory elements in operative linkage with a nucleotide sequence encoding a bioluminescent polypeptide.\",\n \"For example, the recombinant nucleic acid molecule in a vector can contain, in operative linkage, a plant enhancer, plant promoter, a translational enhancer, a nucleotide sequence encoding a bioluminescent polypeptide, a translation termination sequence, and transcription termnination sequence.\",\n \"It should be recognized that one or more of the regulatory elements initially can be a component of the vector or of the recombinant nucleic acid molecule.\",\n \"In one embodiment, a vector of the invention contains a recombinant nucleic acid molecule comprising, in operative linkage, a CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luciferase polypeptide or variant thereof, and a CaMV 35S terminator.\",\n \"In another embodiment, the recombinant nucleic acid molecule contained in the vector comprises, in operative linkage, a dual CaMV 35S enhancer, a CaMV 35S promoter, a TEV translational enhancer, a nucleotide sequence encoding a luc+ luciferase variant, and a CaMV 35S terminator.\",\n \"Vectors of the invention are exemplified herein by a vector having the nucleotide sequence of SEQ ID NO:1 (see, also, FIG.\",\n \"1), and by a vector having the nucleotide sequence of SEQ ID NO:2 (see, also, FIG.\",\n \"2).\",\n \"Additional vectors of the invention are exemplified by those set forth as SEQ ID NO:4 and SEQ ID NO:5.The present invention also provides a host cell containing a recombinant nucleic acid molecule of the invention, or containing a vector of the invention.\",\n \"The present invention further provides a method of producing a genetically modified plant cell that is visibly bioluminescent.\",\n \"As used herein, the term “genetically modified,” when used in reference to a plant cell, means that the plant cell contains an exogenously introduced nucleic acid molecule.\",\n \"For purposes of the present invention, the introduced nucleotide sequence generally is a recombinant nucleic acid molecule comprising one or more regulatory elements operatively linked to a nucleotide sequence encoding a bioluminescent polypeptide, or a vector containing such a recombinant nucleic acid molecule.\",\n \"The nucleic acid molecule can be transiently associated with the plant cell, but generally is stably associated with the genetically modified cell, as well as its progeny, either as an autonomously replicating molecule or due to integration into the plant cell genomic DNA or plastid DNA that is stably maintained in the plant cell.\",\n \"If desired, a transgene comprising a recombinant nucleic acid molecule of the invention can be introduced into a plant genome in a site-specific matter, for example, by homologous recombination.\",\n \"A method of the invention can be performed, for example, by introducing a transgene comprising a recombinant nucleic acid molecule of the invention into a plant cell such that the encoded bioluminescent polypeptide is expressed in the plant cell at a level that can produce at least about 750,000 to 1×106 photons of visible light/mm2/second.\",\n \"As such, the present invention also provides a genetically modified plant cell produced by a method of the invention, as well as to a transgenic plant containing or generated from such a genetically modified plant cell, and to plant tissues, plant cells, plant organs and seeds produced by such a transgenic plant; as well as to a cDNA or genomic DNA library prepared from the transgenic plant, or from a plant cell or plant tissue obtained from the transgenic plant.\",\n \"A method of the invention can further include contacting the plant cell, or the genetically modified plant cell derived therefrom, with an agent that reduces or inhibits pigment synthesis in the plant cell or in a tissue or organ of a transgenic plant of the invention, for example, the synthesis of an anthocyanin, which is a water-soluble red to blue plant pigment; a carotene, which is an orange-yellow pigment located in the chloroplasts; chlorophyll, which is the green pigment in green plants; xanthophyll, which is a yellow to colorless photosynthetic plant pigment, or any other pigment that can absorbs visible light, for example, lycopene, which is a red pigment found in tomatoes.\",\n \"The agent that is selected for reducing or inhibiting pigmentation of a plant is based on the particular pigment, and that the pathway involved in synthesis of the pigment.\",\n \"For example, an agent such as norfluorazon, which inhibits carotenoid synthesis, can be used to reduce the amount of chlorophyll in the plant cell, thus enhancing the emission of a greater number of photons from the plant cell, or from a transgenic plant derived from the plant cell.\",\n \"The term “plant” is used broadly herein to include any plant at any stage of development, or to part of a plant, including a plant cutting, a plant cell, a plant cell culture, a plant organ, a plant seed, and a plantlet.\",\n \"A plant cell is the structural and physiological unit of the plant, comprising a protoplast and a cell wall.\",\n \"A plant cell can be in the form of an isolated single cell or aggregate of cells such as a friable callus, or a cultured cell, or can be part of higher organized unit, for example, a plant tissue, plant organ, or plant.\",\n \"Thus, a plant cell can be a protoplast, a gamete producing cell, or a cell or collection of cells that can regenerate into a whole plant.\",\n \"As such, a seed, which comprises multiple plant cells and is capable of regenerating into a whole plant, is considered plant cell for purposes of this disclosure.\",\n \"A plant tissue or plant organ can be a seed, protoplast, callus, or any other groups of plant cells that is organized into a structural or functional unit.\",\n \"Particularly useful parts of a plant include harvestable parts and parts usefull for propagation of progeny plants.\",\n \"A harvestable part of a plant can be any useful part of a plant, for example, flowers, pollen, seedlings, tubers, leaves, stems, fruit, seeds, roots, and the like.\",\n \"A part of a plant useful for propagation includes, for example, seeds, fruits, cuttings, seedlings, tubers, rootstocks, and the like.\",\n \"A transgenic plant can be regenerated from a genetically modified plant cell, i.e., a whole plant can be regenerated from a plant cell; a group of plant cells; a protoplast; a seed; or a piece of a plant such as a leaf, a cotyledon or a cutting.\",\n \"Regeneration from protoplasts varies from species to species of plants.\",\n \"For example, a suspension of protoplasts can be made and, in certain species, embryo formation can be induced from the protoplast suspension, to the stage of ripening and germination.\",\n \"The culture media generally contains various components necessary for growth and regeneration, including, for example, hormones such as auxins and cytokinins; and amino acids such as glutamic acid and proline, depending on the particular plant species.\",\n \"Efficient regeneration will depend, in part, on the medium, the genotype, and the history of the culture.\",\n \"If these variables are controlled, however, regeneration is reproducible.\",\n \"Regeneration can occur from plant callus, explants, organs or plant parts.\",\n \"Transformation can be performed in the context of organ or plant part regeneration.\",\n \"(see Meth.\",\n \"Enzymol.\",\n \"Vol.\",\n \"118; Klee et al.\",\n \"Ann.\",\n \"Rev.\",\n \"Plant Physiol.\",\n \"38:467 (1987), which is incorporated herein by reference).\",\n \"Utilizing the leaf disk-transformation-regeneration method, for example, disks are cultured on selective media, followed by shoot formation in about two to four weeks (see Horsch et al., supra, 1985).\",\n \"Shoots that develop are excised from calli and transplanted to appropriate root-inducing selective medium.\",\n \"Rooted plantlets are transplanted to soil as soon as possible after roots appear.\",\n \"The plantlets can be repotted as required, until reaching maturity.\",\n \"In vegetatively propagated crops, the mature tralsgenic plants are propagated utilizing cuttings or tissue culture techniques to produce multiple identical plants.\",\n \"Selection of desirable transgenotes is made and new varieties are obtained and propagated vegetatively for commercial use.\",\n \"In seed propagated crops, the mature transgenic plants can be self-pollinated to produce a homozygous inbred plant.\",\n \"The resulting inbred plant produces seeds that contain the introduced transgene, which can comprise a heterologous nucleotide sequence encoding a bioluminescent polypeptide, and can be grown to produce plants that express the polypeptide.\",\n \"As such, the invention provides seeds produced by a transgenic plant obtained by a method of the invention.\",\n \"Transgenic plants comprising different transgenes also can be crossbred, thereby providing a means to obtain transgenic plants containing two or more different transgenes, one of which encodes a bioluminescent polypeptide and the other or others of which confer a desirable characteristic to the plant.\",\n \"Methods for breeding plants and selecting for crossbred plants having desirable characteristics or other characteristics of interest are well known in the art.\",\n \"A method of the invention can be performed by introducing a recombinant nucleic acid molecule of the invention into a plant cell.\",\n \"As used herein, the term “introducing” means transferring a polynucleotide into a plant cell.\",\n \"A nucleic acid molecule can be introduced into a cell by a variety of methods.\",\n \"For example, the recombinant nucleic acid molecule of the invention, which can be contained in a vector, can be introduced into a plant cell using a direct gene transfer method such as electroporation or microprojectile mediated transformation, or using Agrobacterium mediated transformation.\",\n \"As used herein, the term “transformed” refers to a plant cell containing an exogenously introduced nucleic acid molecule.\",\n \"It should be recognized that one or more nucleic acid molecules, which are the same or different and at least one of which is a recombinant nucleic acid molecule of the invention, can be introduced into a plant cell, thereby providing a means to obtain a genetically modified plant cell containing multiple copies of a single transgene, or containing two or more different genes, either or both of which can be present in multiple copies.\",\n \"Such genetically modified plant cells can be produced, for example, by simply selecting plant cells having multiple copies of a single type of transgene; by cotransfecting plant cells with two or more populations of different transgenes and identifying those containing the two or more different transgenes; or by crossbreeding transgenic plants, each of which contains one or more desired transgenes, and identifying those progeny having the desired transgenes.\",\n \"Methods for introducing a nucleic acid molecule into a plant cell to obtain a transformed plant also include direct gene transfer (see European Patent A 164 575), injection, electroporation, biolistic methods such as particle bombardment, pollen-mediated transformation, plant RNA virus-mediated transformation, liposome-mediated transformation, transformation using wounded or enzyme-degraded immature embryos, or wounded or enzyme-degraded embryogenic callus, and the like.\",\n \"Transformation methods using Agrobacterium tumefaciens tumor inducing (Ti) plasmids or A. rhizogenes root-inducing (Ri) plasmids, or other plant virus vectors are well known in the art (see, for example, WO 99/47552; Weissbach and Weissbach, “Methods for Plant Molecular Biology” (Academic Press, NY 1988), section VIII, pages 421-463; Grierson and Corey, “Plant Molecular Biology” 2d Ed.\",\n \"(Blackie, London 1988), Chapters 7-9, each of which is incorporated herein by reference; Horsch et al., supra, 1985).\",\n \"The wild type form of A. tumefaciens, for example, contains a Ti plasmid, which directs production of tumorigenic crown gall growth on host plants.\",\n \"Transfer of the tumor inducing T-DNA region of the Ti plasmid to a plant genome requires the Ti plasmid-encoded virulence genes as well as T-DNA borders, which are a set of direct DNA repeats that delineate the region to be transferred.\",\n \"An Agrobacterium based vector is a modified form of a Ti plasmid, in which the tumor inducing functions are replaced by a nucleotide sequence of interest that is to be introduced into the plant host.\",\n \"Methods of using Agrobacterium mediated transformation include cocultivation of Agrobacterium with cultured isolated protoplasts; transformation of plant cells or tissues with Agrobacterium; and transformation of seeds, apices or meristems with Agrobacterium.\",\n \"In addition, in planta transformation by Agrobacterium can be performed using vacuum infiltration of a suspension of Agrobacterium cells (Bechtold et al., C.R.\",\n \"Acad.\",\n \"Sci.\",\n \"Paris 316:1194, 1993, which is incorporated herein by reference).\",\n \"Agrobacterium mediated transformation can employ cointegrate vectors or binary vector systems, in which the components of the Ti plasmid are divided between a helper vector, which resides permanently in the Agrobacterium host and carries the virulence genes, and a shuttle vector, which contains the gene of interest bounded by T-DNA sequences.\",\n \"Binary vectors are well known in the art (see, for example, De Framond, BioTechnology 1:262, 1983; Hoekema et al., Nature 303:179, 1983, each of which is incorporated herein by reference) and are commercially available (Clontech; Palo Alto Calif.).\",\n \"For transformation, Agrobacterium can be cocultured, for example, with plant cells or wounded tissue such as leaf tissue, root explants, hypocotyls, cotyledons, stem pieces or tubers (see, for example, Glick and Thompson, “Methods in Plant Molecular Biology and Biotechnology” (Boca Raton Fla., CRC Press-1993), which is incorporated herein by reference).\",\n \"Wounded cells within the plant tissue that have been infected by Agrobacterium can develop organs de novo when cultured under the appropriate conditions; the resulting transgenic shoots eventually give rise to transgenic plants, which contain an exogenously introduced circadian-regulated polynucleotide, or an oligonucleotide portion thereof.\",\n \"Agrobacterium mediated transformation has been used to produce a variety of tratsgenic plants, including, for example, transgenic cruciferous plants such as Arabidopsis, mustard, rapeseed and flax; transgenic leguminous plants such as alfalfa, pea, soybean, trefoil and white clover; and transgenic solanaceous plants such as eggplant, petunia, potato, tobacco and tomato (see, for example, Wang et al., “Transformation of Plants and Soil Microorganisms” (Cambridge, University Press 1995), which is incorporated herein by reference).\",\n \"In addition, Agrobacterium mediated transformation can be used to introduce an exogenous nucleic acid molecule into apple, aspen, belladonna, black currant, carrot, celery, cotton, cucumber, grape, horseradish, lettuce, morning glory, muskmelon, neem, poplar, strawberry, sugar beet, sunflower, walnut, asparagus, rice and other plants (see, for example, Glick and Thompson, supra, 1993; Hiei et al., Plant J.\",\n \"6:271-282, 1994; Shimamoto, Science 270:1772-1773, 1995).\",\n \"Suitable strains of A. tumefaciens and vectors as well as transformation of Agrobacteria and appropriate growth and selection media are well known in the art (GV3101, pMK90RK), Koncz, Mol.\",\n \"Gen. Genet.\",\n \"204:383-396, 1986; (C58C1, pGV3850kan), Deblaere, Nucl.\",\n \"Acid Res.\",\n \"13:4777, 1985; Bevan, Nucleic Acid Res.\",\n \"12:8711, 1984; Koncz, Proc.\",\n \"Natl.\",\n \"Acad.\",\n \"Sci.\",\n \"USA 86:8467-8471, 1986; Koncz, Plant Mol.\",\n \"Biol.\",\n \"20:963-976, 1992; Koncz, Specialized vectors for gene tagging and expression studies.\",\n \"In: Plant Molecular Biology Manual Vol.\",\n \"2, Gelvin and Schilperoort (Eds.\",\n \"), Dordrecht, The Netherlands: Kluwer Academic Publ.\",\n \"(1994), 1-22; European Patent A-1 20 516; Hoekema: The Binary Plant Vector System, Offsetdruikkerij Kanters B. V., Alblasserdam (1985), Chapter V; Fraley, Crit.\",\n \"Rev.\",\n \"Plant.\",\n \"Sci., 4:1-46; An, EMBO J.\",\n \"4:277-287, 1985).\",\n \"Where the recombinant nucleic acid molecule is contained in a vector, the vector can contain functional elements, for example “left border” and “right border” sequences of the T-DNA of Agrobacterium, which allow for stable integration into a plant genome.\",\n \"Furthermore, methods and vectors that permit the generation of marker-free transgenic plants, for example, where a selectable marker gene is lost at a certain stage of plant development or plant breeding, are known, and include, for example, methods of co-transformation (Lyznik, Plant Mol.\",\n \"Biol.\",\n \"13:151-161, 1989; Peng, Plant Mol.\",\n \"Biol.\",\n \"27:91-104, 1995), or methods that utilize enzymes capable of promoting homologous recombination in plants (see, e.g., W097/08331; Bayley, Plant Mol.\",\n \"Biol.\",\n \"18:353-361, 1992; Lloyd, Mol.\",\n \"Gen. Genet.\",\n \"242:653-657, 1994; Maeser, Mol.\",\n \"Gen. Genet.\",\n \"230:170-176, 1991; Onouchi, Nucl.\",\n \"Acids Res.\",\n \"19:6373-6378, 1991; see, also, Sambrook et al., supra, 1989).\",\n \"A direct gene transfer method such as electroporation also can be used to introduce a polynucleotide into a cell such as a plant cell.\",\n \"For example, plant protoplasts can be electroporated in the presence of a recombinant nucleic acid molecule comprising a nucleotide sequence encoding a bioluminescent polypeptide, which can be in a vector (Fromm et al., Proc.\",\n \"Natl.\",\n \"Acad.\",\n \"Sci., USA 82:5824, 1985, which is incorporated herein by reference).\",\n \"Electrical impulses of high field strength reversibly permeabilize membranes allowing the introduction of the nucleic acid.\",\n \"Electroporated plant protoplasts reform the cell wall, divide and form a plant callus.\",\n \"Microinjection can be performed as described in Potrykus and Spangenberg (eds.\",\n \"), Gene Transfer To Plants.\",\n \"Springer Verlag, Berlin, N.Y. (1995).\",\n \"A transformed plant cell containing the introduced recombinant nucleic acid molecule can be identified due to the presence of a selectable marker included in the construct, or simply by looking at the plant cells and observing visible bioluminescence.\",\n \"Microprojectile mediated transformation also can be used to introduce a recombinant nucleic acid molecule of the invention into a plant cell (Klein et al., Nature 327:70-73, 1987, which is incorporated herein by reference).\",\n \"This method utilizes microprojectiles such as gold or tungsten, which are coated with the desired nucleic acid molecule by precipitation with calcium chloride, spermidine or polyethylene glycol.\",\n \"The microprojectile particles are accelerated at high speed into a plant tissue using a device such as the BIOLISTIC PD-1000 particle gun (BioRad; Hercules Calif.).\",\n \"Microprojectile mediated delivery (“particle bombardment”) is especially useful to transform plant cells that are difficult to transform or regenerate using other methods.\",\n \"Methods for the transformation using biolistic methods are well known (Wan, Plant Physiol.\",\n \"104:37-48, 1984; Vasil, BioTechnology 11:1553-1558, 1993; Christou, Trends in Plant Science 1:423-431, 1996).\",\n \"Microprojectile mediated transformation has been used, for example, to generate a variety of transgenic plant species, including cotton, tobacco, corn, hybrid poplar and papaya (see Glick and Thompson, supra, 1993).\",\n \"Important cereal crops such as wheat, oat, barley, sorghum and rice also have been transformed using microprojectile mediated delivery (Duan et al., Nature Biotech.\",\n \"14:494-498, 1996; Shimamoto, Curr.\",\n \"Opin.\",\n \"Biotech.\",\n \"5:158-162, 1994).\",\n \"A rapid transformation regeneration system for the production of transgenic plants such as a system that produces transgenic wheat in two to three months (see European Patent No.\",\n \"EP 0709462A2, which is incorporated herein by reference) also can be useful for producing a transgenic plant according to a method of the invention, thus allowing more rapid identification of gene functions.\",\n \"The transformation of most dicotyledonous plants is possible with the methods described above.\",\n \"Transformation of monocotyledonous plants also can be transformed using, for example, biolistic methods as described above, protoplast transformation, electroporation of partially permeabilized cells, introduction of DNA using glass fibers, Agrobacterium mediated transformation, and the like.\",\n \"Plastid transformation also can be used to introduce a nucleic acid molecule into a plant cell (U.S. Pat.\",\n \"Nos.\",\n \"5,451,513, 5,545,817, and 5,545,818; WO 95/16783; McBride et al., Proc.\",\n \"Natl.\",\n \"Acad.\",\n \"Sci., USA 91:7301-7305, 1994).\",\n \"Chloroplast transformation involves introducing regions of cloned plastid DNA flanking a desired nucleotide sequence, for example, a selectable marker together with polynucleotide of interest into a suitable target tissue, using, for example, a biolistic or protoplast transformation method (e.g., calcium chloride or PEG mediated transformation).\",\n \"One to 1.5 kb flanking regions (“targeting sequences”) facilitate homologous recombination with the plastid genome, and allow the replacement or modification of specific regions of the plastome.\",\n \"Using this method, point mutations in the chloroplast 16S rRNA and rps12 genes, which confer resistance to spectinomycin and streptomycin and can be utilized as selectable markers for transformation (Svab et al., Proc.\",\n \"Natl.\",\n \"Acad.\",\n \"Sci., USA 87:8526-8530, 1990; Staub and Maliga, Plant Cell 4:39-45, 1992), resulted in stable homopiasmic transformants, at a frequency of approximately one per 100 bombardments of target leaves.\",\n \"The presence of cloning sites between these markers allowed creation of a plastid targeting vector for introduction of foreign genes (Staub and Maliga, EMBO J.\",\n \"12:601-606, 1993).\",\n \"Substantial increases in transformation frequency are obtained by replacement of the recessive rRNA or r-protein antibiotic resistance genes with a dominant selectable marker, the bacterial aadA gene encoding the spectinomycin-detoxifying enzyme aminoglycoside-3′-adenyltransferase (Svab and Maliga, Proc.\",\n \"Natl.\",\n \"Acad.\",\n \"Sci., USA 90:913-917, 1993).\",\n \"Approximately 15 to 20 cell division cycles following transformation are generally required to reach a homoplastidic state.\",\n \"Plastid expression, in which genes are inserted by homologous recombination into all of the several thousand copies of the circular plastid genome present in each plant cell, takes advantage of the enormous copy number advantage over nuclear-expressed genes to permit expression levels that can readily exceed 10% of the total soluble plant protein.\",\n \"Plants suitable for treatment according to a method of the invention so as to be visibly bioluminescent can be monocots or dicots and include, but are not limited to, maize, wheat, barley, rye, sweet potato, bean, pea, chicory, lettuce, cabbage, cauliflower, broccoli, turnip, radish, spinach, asparagus, onion, garlic, pepper, celery, squash, pumpkin, hemp, zucchini, apple, pear, quince, melon, plum, cherry, peach, nectarine, apricot, strawberry, grape, raspberry, blackberry, pineapple, avocado, papaya, mango, banana, soybean, tomato, sorghum, sugarcane, sugar beet, sunflower, rapeseed, clover, tobacco, carrot, cotton, alfalfa, rice, potato, eggplant, cucumber, Arabidopsis thaliana, and woody plants such as coniferous and deciduous trees.\",\n \"Thus, a transgenic plant or genetically modified plant cell of the invention can be an angiosperm or gymnosperm.\",\n \"Angiosperms are divided into two broad classes based on the number of cotyledons, which are seed leaves that generally store or absorb food; a monocotyledonous angiosperm has a single cotyledon, and a dicotyledonous angiosperm has two cotyledons.\",\n \"Angiosperms produce a variety of useful products including materials such as lumber, rubber, and paper; fibers such as cotton and linen; herbs and medicines such as quinine and vinblastine; ornamental flowers such as roses and, where included within the scope of the present invention, orchids; and foodstuffs such as grains, oils, fruits and vegetables.\",\n \"Angiosperms encompass a variety of flowering plants, including, for example, cereal plants, leguminous plants, oilseed plants, hardwood trees, fruit-bearing plants and ornamental flowers, which general classes are not necessarily exclusive.\",\n \"Cereal plants, which produce an edible grain cereal, include, for example, corn, rice, wheat, barley, oat, rye, orchardgrass, guinea grass, sorghum and turfgrass.\",\n \"Leguminous plants include members of the pea family (Fabaceae) and produce a characteristic fruit known as a legume.\",\n \"Examples of leguminous plants include, for example, soybean, pea, chickpea, moth bean, broad bean, kidney bean, lima bean, lentil, cowpea, dry bean, and peanut, as well as alfalfa, birdsfoot trefoil, clover and sainfoin.\",\n \"Oilseed plants, which have seeds that are useful as a source of oil, include soybean, sunflower, rapeseed (canola) and cottonseed.\",\n \"Angiosperms also include hardwood trees, which are perennial woody plants that generally have a single stem (trunk).\",\n \"Examples of such trees include alder, ash, aspen, basswood (linden), beech, birch, cherry, cottonwood, eln, eucalyptus, hickory, locust, maple, oak, persimmon, poplar, sycamore, walnut, sequoia, and willow.\",\n \"Trees are useful, for example, as a source of pulp, paper, structural material and fuel.\",\n \"Angiosperms are fruit-bearing plants that produce a mature, ripened ovary, which generally contains seeds.\",\n \"A fruit can be suitable for human or animal consumption or for collection of seeds to propagate the species.\",\n \"For example, hops are a member of the mulberry family that are prized for their flavoring in malt liquor.\",\n \"Fruit-bearing angiosperms also include grape, orange, lemon, grapefruit, avocado, date, peach, cherry, olive, plum, coconut, apple and pear trees and blackberry, blueberry, raspberry, strawberry, pineapple, tomato, cucumber and eggplant plants.\",\n \"An ornamental flower is an angiosperm cultivated for its decorative flower.\",\n \"Examples of commercially important ornamental flowers include rose, lily, tulip and chrysanthemum, snapdragon, camellia, carnation and petunia plants, and can include orchids.\",\n \"The skilled artisan will recognize that the methods of the invention can be practiced using these or other angiosperms, as desired, as well as gymnosperms, which do not produce seeds in a fruit.\",\n \"As such, the methods of the invention can be used to prepare bioluminescent plants of various families, including, for example, Asteraceae-Aster family; Fabaceae-Pea family; Rubiaceae-Madder family; Poaceae-Grass family; Euphorbiaceae-Spurge family; Lamiaceae-Mint family, Melastomataceae-Melastome family; Liliaceae-Lily family; Scrophulariaceae-Figwort family; Acanthaceae-Ancanthus family, Myaceae-Myrtle family, Cyperaceae-Sedge family; Ericaceae-Heath family, Apiaceae-Carrot family, Rosaceae-Rose family; Brassicaceae-Mustard family, Araceae-Arum family, Asclepiadaceae-Milkweed family; Arecaceae-Pahm family, Solanaceae-Potato family, Boraginaceae-Borage family; Gesneriaceae-Generiad family, Lauraceae-Laurel family, and Bromeliaceae-Bromeliad family.\",\n \"In certain embodiments, the methods of the invention also can be used to prepare bioluminescent plants of Orchidaceae-Orchid family, whereas in other embodiments, members of the orchid family are specifically excluded.\",\n \"A recombinant nucleic acid molecule of the invention or a vector containing the recombinant nucleic acid molecule provides a valuable reagent for identifying plant cells containing an introduced nucleic acid molecule.\",\n \"For example, the recombinant nucleic acid molecule, or vector containing the recombinant nucleic acid molecule, can include a nucleotide sequence of interest.\",\n \"Following transformation of plant cells with the recombinant nucleic acid molecule of the invention, which contains the nucleotide sequence of interest, visibly bioluminescent plant cells can be selected simply by looking at the plant cells, for example, under a microscope, and picking out the ones that are visibly bioluminescent, thereby selecting plant cells that also contain the nucleotide sequence of interest.\",\n \"Thus, a recombinant nucleic acid molecule of the invention is useful as a selectable marker, and provides a quick, efficient, and inexpensive means to identify a plant cell containing a nucleotide sequence of interest.\",\n \"A nucleotide sequence of interest also can encode an immunogenic polypeptide, for example, a viral or bacterial polypeptide, or a peptide portion thereof.\",\n \"A genetically modified plant cell, particularly a transgenic plant comprising such a plant cell, can be used as a source of a vaccine, wherein the individuals to be immunized merely ingest the plant, and a protective or palliative immune response is obtained.\",\n \"As an additional advantage, problems generally associated with the preparation and purification of a vaccine, as well as storage and refrigeration problems and the like are obviated.\",\n \"In view of these examples, it will be recognized that genetically modified plant cells and transgenic plant comprising such cells containing essentially any nucleotide sequence of interest can be prepared, and that the inclusion of a recombinant nucleic acid molecule of the invention provides a selectable marker that facilitates the identification of a transformed plant cell or plant comprising such a cell that contains the nucleotide sequence of interest A recombinant nucleic acid molecule of the invention, or a vector containing such a nucleic acid molecule, also is useful for preparing transgenic plants having great ornamental value.\",\n \"Visibly bioluminescent plants containing genetically modified plant cells have not previously been described.\",\n \"As such, the transgenic plants of the invention, which “glow in the dark,” provide a novel ornamental plant.\",\n \"For example, a transgenic plant expressing a luciferase polypeptide or variant thereof can be sprayed with water containing luciferin, or the transgenic plant or a cutting or other portion of the transgenic plant, for example, an inflorescence of the plant, can be dipped or placed in a solution containing luciferin, whereupon the visible bioluminescence is produced.\",\n \"For example, flowers can be cut from a transgenic plant of the invention and placed in water containing luciferin, wherein the luciferin is taken up and transported to the flower, thus providing a visibly bioluminescent flower.\",\n \"Generally, the amount of luciferin sufficient to induce visible bioluminescence includes at least about 0.1 mM, and concentrations from about 0.1 mM to 5 mM or more can be used.\",\n \"In addition, the solution containing luciferin can further include a surfactant, for example, a non-ionic detergent such as TRITON X-100 detergent or NONIDET P40 detergent in a concentration of about 0.001% to about 0.1%, thus facilitating entry of the luciferin into the plant cells.\",\n \"Significantly, luciferase-mediated bioluminescence continues for several hours following a single administration of luciferin and, therefore, there is no requirement that the plants be continually treated to maintain bioluminescence.\",\n \"Furthermore, the transgenic plants can be selected based on their expression of variable levels of light, including plants that glow dimly, yet visibly, and those that glow very brightly.\",\n \"In addition to providing general ornamental value, the visibly bioluminescent transgenic plants of the invention also can provide a practical value.\",\n \"For example, visibly bioluminescent transgenic plants that grow as bushes, shrubs or trees can be arranged along a walkway or driveway.\",\n \"Where such plants express luciferase or a variant thereof, for example, the plants can be watered in the early evening with a solution containing luciferin such that they illuminate the walkway or driveway after night falls.\",\n \"Thus, the transgenic plants of the invention provide the additional advantage they can be used to make an area safer by illuminating the area.\",\n \"The present invention also relates to kit, which contains a genetically modified plant cell of the invention, or a derivative of the genetically modified plant cell, for example, a transgenic plant derived from the genetically modified plant cell, or a cell, a tissue, or an organ of such a transgenic plant.\",\n \"As such, a kit of the invention can contain one or more flowers, bracts, leaves or other tissues or organs, which, upon contact with luciferin, are visibly luminescent.\",\n \"Accordingly, a kit of the invention can further include an amount of luciferin sufficient for generating visible bioluminescence of the genetically modified plant cell or the derivative of the genetically modified plant cell, and also can include a plurality of such amounts of luciferin, thus allowing the generation of visible bioluminesce a number of times, as desired.\",\n \"In addition, a kit of the invention can include an amount of a carotenoid inhibitor, for example, norfluorazon, sufficient for reducing or inhibiting chlorophyll production in the genetically modified plant cell or the derivative of the genetically modified plant cell.\",\n \"In another embodiment, a kit of the invention contains one or more cuttings, seeds, or other portion or derivative of a visibly bioluminescent plant of the invention such that a visibly luminescent transgenic plant can be grown therefrom.\",\n \"As such, the kit also can include reagents for growing a transgenic plant from the cutting or seed, including, for example, a suitable plant food or other nutrient source required for growth of the particular plant.\",\n \"In addition, the kit can include an amount of a carotenoid inhibitor sufficient for reducing or inhibiting chlorophyll production in a transgenic plant grown from the cutting or seed; and/or can include an amount of luciferin sufficient for generating visible bioluminescence of a transgenic plant grown from the cutting or seed.\",\n \"Generally, a kit of the invention includes a container for the components of the kit.\",\n \"It should be recognized, however, that the container can be any convenient means for holding a component of the kit, including, for example, a packet or collection of packets containing seeds obtained from a transgenic plant of the invention, or containing predetermined amounts of a reagent such as luciferin, which can be added to a specified amount of water to provide a solution sufficient to generate visible bioluminescence from a composition of the invention.\",\n \"In one embodiment, the kit comprises a flower cutting of a transgenic plant of the invention in a vase, and can further include one or more predetermined amounts of a reagent such as luciferin.\",\n \"In another embodiment, the kit comprises a seed or a cutting obtained from a transgenic plant of the invention, and further includes a reagent such as luciferin and/or norfluorazon, in which the seed or cutting, or a plant derived therefrom, can be grown.\",\n \"The following examples are intended to illustrate but not limit the invention.\",\n \"EXAMPLE 1 Preparation of Luciferase Expression Vectors This example provides a method for preparing luciferase constructs that can be incorporated into an expression vector and are useful for producing a visibly bioluminescent plant.\",\n \"Plasmid pRTL2, which contains a CaMV 35S promoter with duplicated enhancer, a TEV leader and initial coding sequence from pTL-7SN, and a CaMV 35S polyadenylation (poly A) signal sequence (Topfer et al., Nucl.\",\n \"Acids Res.\",\n \"15:5890, 1987; Restrepo et al., Plant Cell 2:987, 1990, each of which is incorporated herein by reference; see, also, Carrington and Freed, supra, 1990) was sequentially digested with Nco I and Xba I, then separated by electrophoresis on a 0.8% agarose gel.\",\n \"The nucleotide sequence of the CaMV 35S promoter with duplicated enhancer, TEV leader and initial coding sequence, and CaMV 35S poly A signal sequence of pRTL2 is shown as SEQ ID NO:3.A DNA band of approximately 3.9 kb representing the linear pRTL2 (see nucleotides 1 to 1188 and 2844 to 5475 of SEQ ID NO:1) was isolated and extracted from the gel using a QIAQUICK DNA gel extraction kit (Qiagen; Valencia Calif.) according to manufacturer's instructions.\",\n \"The Nco I site in the TEV leader contains the start codon for translation.\",\n \"Translational fusions containing the initial 20 amino acid residues of the TEV polyprotein are made by ligation to one of the polylinker sites, provided care is taken to avoid the STOP codon in the Xba I site (see SEQ ID NO:3).\",\n \"In a parallel reaction, pGL3 (Promega Corp., Madison Wis.), which contains a modified luciferase coding sequence, luc+ (Groskreutz et al., Promega Notes 50:2, 1995; U.S. Pat.\",\n \"No.5,670,356, each of which is incorporated herein by reference) was digested sequentially with Xba I and Nco I, then separated by electrophoresis on a 0.8% agarose gel.\",\n \"Two bands were present—a 1.6 kb band representing the luc+ coding sequence (see U.S. Pat.\",\n \"No.5,670,356) and an approximately 3 kb band representing the remainder of the pGL3 plasmid.\",\n \"The approximately 1.6 kb luc+ band (see nucleotides 1188 to 2844of SEQ ID NO:1) was isolated and extracted as described above.\",\n \"The isolated luc+ sequence (4 μl) was ligated into the linearized pRTL2 vector (1 μl) using ligase and ligation buffer from BRL (Gaithersburg Md.).\",\n \"The ligation was performed overnight at 16° C. E. coli DH5α cells were transformed using a BioRad Electroporator in a ratio of 2 μl of ligated DNA to 20 μl of cells.\",\n \"Following electroporation, the transformed cells were incubated in LB medium for 1 hr at 37° C., then were plated on LB plates containing 50 μg/ml ampicillin.\",\n \"Of approximately 3000 colonies, 32 were selected and screened by PCR for the luc+ insert.\",\n \"PCR primers were specific for the 5′ end of the TEV promoter in pRTL2 and for the 3′ end of the poly A terminator in pRTL2 (see SEQ ID NO:3).\",\n \"PCR analysis revealed that 30 of the colonies contained the luc+ insert.\",\n \"The presence of the insert in 6 clones was verified by restriction analysis, and two candidate clones, designated TK1-7 and TK1-8, were sequenced to confirm the presence of the luc+ insert.\",\n \"The TK1-8 clone was digested at 37° C. for 2 hr with Hind III, and the fragment containing the CaMV 35S promoter with duplicated enhancer, TEV leader, luc+ coding sequence and poly A termination signal (designated “TK1829”; see FIG.\",\n \"2, and nucleotides 8366 to 11,113 of SEQ ID NO:2) was isolated by electrophoresis as described above.\",\n \"The binary vector, pPZP221, also was digested with Hind III and isolated.\",\n \"The linearized pPZP221 vector was treated with shrimp alkaline phosphatase for 1 hr at 37° C., to remove terminal phosphate groups, followed by 15 min at 65° C. to heat inactivate the enzyme, then the TK1829 insert and linearized pPZP221 vector were ligated and transformed into DH5α cells as described above.\",\n \"Cells were plated on LB agar plates containing X-gal, IPTG, and 100 μg/ml spectinomycin.\",\n \"Forty-six white colonies were selected, and the presence of the luc+ coding sequence was confirmed by PCR as described above; all were positive for the luc+ sequence.\",\n \"Six samples were selected and the orientation of the insert was determined by restriction digest analysis, using Sca I or Nar I.\",\n \"One construct (#29) having the correct orientation was selected.\",\n \"The binary vector containing the luc+ insert was designated pPZPTK1829 (FIG.\",\n \"2; SEQ ID NO:2), and the inserted expression construct was designated TK1829 (nucleotides 8366 to 11,113 of SEQ ID NO:2).\",\n \"TK1829 was sequenced using primers specific for 5′ and 3′ insert flanking regions of the pPZP221 vector.\",\n \"The pRTL2 vector from which the TK1829 insert was obtained was designated pTK1829 (see FIG.\",\n \"1; see, also, SEQ ID NO:1).\",\n \"The pMT-OM-LUC+ (SEQ ID NO:4; FIG.\",\n \"3) and pACT-OM-LUC+ (SEQ ID NO:5; FIG.\",\n \"4) binary vector plasmids were prepared from the pPZP221 binary vector, as described above.\",\n \"The metallothionein (MT) or actin (ACT) promoter was inserted immediately preceding the omega enhancer in a Bam HI site, which provides a convenient site to substitute other transcriptional regulatory elements.\",\n \"EXAMPLE 2 Preparation and Characterization of Visibly Bioluminescent Plants This example provides a method for introducing a luciferase expression construct into plants such that a genetically modified visibly bioluminescent plant is produced, and a method for determining a threshold value of photon emission required for visible luminescence.\",\n \"A.\",\n \"Preparation of TK1829 Transformed Plants The TK1829 expression construct was introduced from pPZPTK1829 into Agrobacterium l strain ABI 1 with the help of pRK2013 by tri-parental mating.\",\n \"Agrobacterium containing the TK1829 construct were selected based on their resistance to the 100 μg/ml spectinomycin, 50 μg/ml kanamycin, and 25 μg/ml chloramphenicol.\",\n \"TK1829-Agrobacterium strains were grown, then DNA was purified and checked by restriction digest analysis using Sca I (2 hr; 37° C.) to verify the presence of the insert.\",\n \"Briefly, TK1829-Agrobacterium was grown as described below, and 28 day old plants were infiltrated.\",\n \"Supernova (Snv) strain plants and Columbia (wild type) plants were each infiltrated.\",\n \"Two sets of infiltrations were performed on Columbia and Snv plants.\",\n \"Each time, two 4 inch pots of each line were used and each pot contained approximately 50 plants, for a total of 200 plants per line infiltrated.\",\n \"For in planta plant transformation, 4 inch pots were prepared with Scott's Metro mix 200 soil and covered with window screen mesh.\",\n \"Soil was wet and mounded up, and the screen mesh was secured by wrapping a rubber band around the neck of the pot, maling sure that the mesh was in contact with the soil surface so that germinating seedlings could get through the mesh.\",\n \"Seeds were stratified in 10 to 50 ml of sterile water and stored at 4° C. for 3 days prior to being applied to pots.\",\n \"Seeds were sown using a dropper, to spread seeds evenly over the surface of the mesh.\",\n \"Plants were grown at approximately 22° C., in 12 hr light:12 hr dark per day to a stage at which bolts were anywhere from 3 to 10 inches high.\",\n \"The bolts were cut off to induce growth of secondary inflorescence.\",\n \"Infiltration was most effective 4 to 5 days after decapitation of bolts, but can be performed up to 8 days later.\",\n \"For infiltration, 5 to 20 ml liquid preculture of Agrobactenium containing the appropriate construct were grown for about 2 days in a shaking incubator at 28° C., then 400 ml of LB was inoculated with the preculture and incubate for one additional day.\",\n \"The large culture generally was prepared the day before the infiltration was performed.\",\n \"Transformed Agrobacterium cells were grown to an optical density at 600 nm (OD600) of approximately 1.2, then harvested by centrifugation (4000 rpm, 10 min, room temperature), and resuspended in infiltration medium (1/2×Murashige and Skoog salts; 1×Gamborg's vitamins; 5.0% sucrose; 0.44 μM benzylamino-purine (10 μg per liter of a 1 ng/ml stock); pH adjusted to 5.7 using KOH; then add 200 μl/l Silwet; media need not be sterilized if used immediately) to an OD 600 of approximately 0.8.The OD600 of an overnight culture generally was about 1.2 to 1.6 units.\",\n \"A 400 ml suspension generally was adequate for an infiltration procedure.\",\n \"The same suspension can be used as many as three times.\",\n \"About 150-200 ml of the Agrobacterium suspension was added to either a beaker or dish and the prepared plants were inverted into the suspension.\",\n \"The plants were entirely submerged and kept in the solution for 10 to 15 min.\",\n \"The pots then were removed from the suspension and drained thoroughly, laid on their side in a plastic flat, and covered with a plastic dome or Saran Wrap to maintain humidity.\",\n \"The flat was uncovered the next day and the pots set upright.\",\n \"A little water was added to the bottom of the tray to help keep the humidity level high while the plants were recovering.\",\n \"The plants were grown for 3 to 4 weeks, keeping the bolts from each pot together and separated from neighboring pots and different infiltrations.\",\n \"Once plants began to yellow and senesce, watering was reduced or discontinued.\",\n \"Seeds were harvested together from one pot.\",\n \"Transformants were selected on 150×15mm selection plates (1/2×Murashige and Skoog salts; 1×Gamborg's vitamins; 1% sucrose; 0.5 g/l MES; 0.8% agarose; pH to 5.7; autoclaved, then add 75 μg/ml gentamycin, 500 μg/ml carbenicilin, 50 μg/ml kanamycin).\",\n \"Seeds were surface sterilize for 2 min in 500 μl 70% ethanol; followed by 10 min in 40% bleach, 1% SDS, 59% water, then rinsed 3 times with sterile water.\",\n \"The seeds were resuspended in 3 to 4 ml of sterile water and plated by pouring onto a plate and shaking until evenly distributed.\",\n \"The plates were allowed to sit for 5 to 10 min, then excess liquid was removed.\",\n \"Approximately 70 to 100 μl of dry seeds were added per plate.\",\n \"Plants were allowed to stratify at 4° C. for 3 days, then transferred to a growth chamber.\",\n \"In some experiments, the herbicide, norflurazon (5 μM; 4-chloro-5-(methylamino)-2-{3-(tri-fluoromethyl)phenyl}-3(2H)-pyridazinone) was included in the growth medium.\",\n \"Norflurazon inhibits chlorophyll synthesis, resulting in the growth of “white” plants.\",\n \"Treatment of plants with norflurazon resulted in greater emission of visible bioluminescence due to the lack of chlorophyll, which absorbs light and blocks light emission (see Tables).\",\n \"After 7 to 10 days (depending on the antibiotic resistance selected for), transformants were identified as dark green plants with elongated roots that penetrated the agar.\",\n \"These seedlings were transferred onto a second set of selection plates.\",\n \"After several days, plants had 4 to 8 true leaves and branching roots.\",\n \"Pseudo-resistant plants were unable to survive in this medium.\",\n \"The plants then were transferred to soil, and kept covered for the first few days.\",\n \"Plants were allowed to recover, bolt, set seed and senesce.\",\n \"Seeds were harvested when dry, then were further dried in a dessicator for 1 week.\",\n \"Yield was approximately 0.5 to 0.75 ml of seed per pot.\",\n \"T1 (transgenic first generation) seeds in the Columbia background were screened on 75 μg/ml gentamycin and 500 μg/ml carbenicilin MS plates.\",\n \"T1 seeds in the Snv background were screened on 75 μg/ml gentamycin, 500 μg/ml carbenicilin, and 50 μg/ml kanamycin (Snv strain contains a transgene conferring kanamycin resistance).\",\n \"Over an 8 week period, 1.6 ml (approximately 32,000 seeds) of each T1 seed set was sterilized and planted in 200 μl (4000 seeds) aliquots on 50 ml plates as described above.\",\n \"Plants were stratified for 3 days at 4° C., then moved to a growth chamber set for 12 hr light: 12 hr dark and approximately 27° C. Sixty-seven T1 seedlings in the Snv background and 25 T1 seedlings in the Columbia background grew under the selection conditions, and were recovered and transferred to soil to harvest T2 seed.\",\n \"It took approximately 12 to 16 weeks to take T1 seed to T2 seed, and varied between plants.\",\n \"B.\",\n \"Characterization of TK1829 Transformed Plants T2 seeds of Columbia and Supernova were sterilized and plated as above, and screened for antibiotic resistance.\",\n \"In addition, the seedlings were screened for bioluminescence, visually and with the Hamamatsu photon counting camera.\",\n \"T2 seedlings within individual families were selected based on their intensity and brightness.\",\n \"Three lines in the Snv background (Snv 11, Snv 37 and Snv 57) and two lines in the Columbia background (Col 10 and Col 19) were selected as the most visibly bioluminescent plants.\",\n \"Seventy-eight T3 individuals of Snv 11, which was extremely bright, were selected based on their visible bioluminescence.\",\n \"Three of these (Snv 11-8, Snv 11-48, Snv 11-52) were selected as the brightest and were maintained as individual lines.\",\n \"For characterization of luciferase expression, tissue was collected from 22 day old plants grown under the same conditions.\",\n \"Examination of Columbia wild-type plants, Columbia plants transformed with 35S:Luc (“35S:Luc”) and Snv plants transformed with cab2:Luc (“Supernova”; see Hicks et al., supra, 1996) was included for comparison.\",\n \"In various assays (see below), protein concentration was determined, relative light units (RLU) were acquired using a luminometer, western blot analysis was performed using recombinant luciferase antibodies, plants were imaged using a Night Owl Camera (see below) to obtain counts of light/mm2/sec, and plants were inspected visually to confirm that bioluminescence was visible to the eye.\",\n \"Based on these assays, the approximate amounts of luciferase per microgram of total protein and the fold increase in enzyme activity were determined, and the amount of light emitted by the plants was determined.\",\n \"For protein assays, a few hundred 22 day old seedlings from plants grown under the same conditions were placed in liquid nitrogen, ground into a fine power, and stored in aliquots of approximately 200 μl volume on dry ice or at −80° C. until the time proteins were extracted.\",\n \"Approximately 100 mg of the powdered tissue was extracted in 500 μl of 1×Promega Cell Lysis Reagent (CLR), ground briefly with a mortar and pestle, then centrifuged at 4° C. for 5 min 14,000 rpm.\",\n \"The supernatant was divided into 100 μl aliquots and transferred immediately to dry ice.\",\n \"Any samples not used within 1 hr were stored at −80° C. to preserve enzyme activity.\",\n \"Protein concentrations were determined for 20 μl of tissue extract using the DC Protein Assay Reagent (BioRad); optical density (OD) was determined at 750 nm using a Pharmacia Ultrospec III spectrophotometer.\",\n \"Acquired Relative Light Units (RLU) were obtained using a Turner TD20E Luminometer.\",\n \"Samples from plants containing the TK1829 construct were diluted 1/100 (5 μl/500 μl) in order for the reading to be within the range of the luminometer.\",\n \"Five μl of sample and 50 μl of Promega Luciferase Assay Reagent (LAR) was used.\",\n \"The maximum value of the range of readings taken every 30 sec for the first 5 min of the reaction was entered into the data tables.\",\n \"The amount of luciferase was approximated by western blot analysis (Tables 1 and 2) or using the RLU data obtained by luminometry and the total protein concentration obtained by spectrophotometry (Table 4).\",\n \"Western blot analysis of the extracts was performed using enhanced chemifluoresence (ECF; Amersham) to visualize protein bands.\",\n \"For samples containing the TK1829 construct, 0.1 μg of total protein was used.\",\n \"For all other samples, 1.0 μg of total protein was used.\",\n \"The 10-fold difference was necessary to keep the luciferase protein concentrations within the range of the assay.\",\n \"A luciferase primary antibody and an alkaline phosphatase secondary antibody (Promega) were used to identify protein bands.\",\n \"Proteins were separated by electrophoresis on, 10% SDS PAGE gels (BioRad), then transferred to Hybond nitrocellulose.\",\n \"Images were visualized using a Molecular Dynamics Fluorimeter, and data was analyzed using ImageQuant software.\",\n \"Luciferase was expressed in 35S:Luc transformed Columbia plants (“35S:Luc”) and cab2:Luc transformed Supernova plants (“Supernova”; Table 1), but these plants were not visibly bioluminescent when examined by eye.\",\n \"In comparison, the TK1829 transformed Columbia plants were visibly bioluminescent.\",\n \"These results indicate that the threshold for visible bioluminescence is between about 85 and 125 pg luc+ protein/μg protein as determined by western blot analysis (Table 1).\",\n \"The TK1829 transformed Supernova plants expressed about 6.5-fold more luc+ than the TK1829 transformed Columbia plants as determined by western blot analysis (Table 1).\",\n \"In addition, the Snv-TK1829 plants emitted about 7-fold more photons of light/mm2/sec than the Col-TK1829 plants.\",\n \"These results demonstrate that visible bioluminescence can be produced in various Arabidopsis strains.\",\n \"In addition, growth of the TK1829 transformed Supernova plants in norflurazon resulted in an almost 2-fold increase in luciferase protein as measured by luminometry, and a greater than 7-fold amount of light emitted as measured using the Night Owl imaging system.\",\n \"This result demonstrates that the amount of visible bioluminescence can be increased by inhibiting chlorophyll synthesis in the transgenic plants.\",\n \"TABLE 1 pg Luciferase Plant Line protein/μg total protein a) Columbia Wild Type 0.01 b) 35S:Luc transformed Columbia 7.0 c) cab2:Luc transformed Supernova 85.0 d) TK1829 transformed Columbia 125.0 e) TK1829 transformed Supernova 834.0 f) TK1829 transformed Supernova with 1456.0 Norflurazon A comparison of western blot data on various plant extracts allowed a determination of the relative difference in luciferase protein concentration among the various plant lines (Table 2).\",\n \"Western blot analysis confirmed that Arabidopsis transformed with the TK1829 construct expressed a much greater amount of luciferase expression as compared to plants transformed with a 35S:Luc construct (see Table 2b and d) and as compared to cab2:Luc-transformed Snv (“Supernova”; see Table 2, c and e).\",\n \"The various plant lines also were examined by imaging analysis using the EGG Berthold Night Owl imaging system (see, also, Example 2C, below).\",\n \"Plants were sprayed with approximately 1 ml of 5 mM luciferin in a solution of 0.001% TRITON X-100 detergent.\",\n \"The camera settings were kept consistent for all plant lines.\",\n \"However, the time duration for taking the images was varied to accommodate the different levels of bioluminescence in the various plant lines.\",\n \"All data was processed and averaged for photon counts of light/area over the time the image was taken (mm/sec); variation between individual plants was eliminated by analyzing pools of plants.\",\n \"TABLE 2 Fold increase in luciferase Plant Comparison protein concentration a) Supernova vs. 35S:Luc 5 b) TK1829 transformed Columbia 128 vs. 35S:Luc c) TK1829 transformed Columbia vs. 31 Supernova d) TK1829 transformed Supernova vs. 244 35S:Luc e) TK1829 transformed Supernova vs. 53 Supernova f) TK1829 transformed Supernova with 2 Norflurazon vs. the same line without g) TK1829 transformed Supernova with 450 Norflurazon vs. 35S:Luc In parallel with the levels of luc+ expression as disclosed above, light emission by the TK1829 transformed plants was substantially greater than that of Columbia plants transformed with the 35S:Luc construct (“35S:Luc”) or of Snv plants transformed with a cab2:Luc construct (“Supernova”; see Table 3).\",\n \"Light emission of Arabidopsis plants transformed with TK1829, as measured by photon counts per area per second, was increased by as much as 5,200 fold over 35S:Luc transformed Columbia plants (see Table 3g).\",\n \"Plants also were inspected by eye to confirm that they were visibly bioluminescent.\",\n \"The TK1829 transformed Columbia and TK1829-transformed Supernova plants were visibly bioluminescent (whether grown in the presence or absence of norflurazon), whereas the none of the other plants, including the 35S:Luc transformed Columbia and cab2:Luc transformed Supernova plants, were visibly bioluminescent.\",\n \"These results demonstrate that transformation of a plant with a construct that can express about 125 pg luc+/μg protein as determined by western blot analysis can effectively generate visibly bioluminescent plants.\",\n \"TABLE 3 Fold increase in light Plant Comparison (cts/mm2/sec) a) Supernova vs. 35S:Luc 70 b) Tk1829 transformed Columbia vs. 35S:Luc 106 c) Tk1829 transformed Columbia vs. 1.5 Supernova d) Tk1829 transformed Supernova vs. 712 35S:Luc e) TK1829 transformed Supernova vs. 10 Supernova f) TK1829 transformed Supernova with 7 Norflurazon vs. Supernova g) TK1829 transformed Supernova with 5,200 Norflurazon vs. 35S:Luc C. Threshold Photon Emission for Visible Bioluminescence In order to provide a standard for determining the threshold number of visible light photons required for visible bioluminescence, the number of photons of visible light emitted per square millimeter per second was determined using a defined system.\",\n \"A Night Owl camera with WinLight Program LB981 (EG&G Berthold; Bad Wildbad, Germany) was used, and all images were acquired under the following conditions: Cosmic Suppression, ON; Defect Correction, ON; Star Killer, OFF; Camera Readout, SLOW; Gain, HIGH; Look up Table set to GAMMA.\",\n \"Measurements were taken in a photography dark room with a plate-to-lens distance of 275 mm (standard setting on Night Owl camera).\",\n \"Each plant was imaged for a time appropriate to the amount of bioluminescence emitted and values were normalized to photon counts/area/second.\",\n \"Approximate photons/mm2/sec were calculated using a conversion of discharges per minute divided by counts per minute.\",\n \"The Night Owl camera system was calibrated using manufacturer-provided standards from the Beckman LS6500 scintillation system.\",\n \"Two standards were used.\",\n \"The first standard contained scintillant alone, which represents background levels of discharges per minute (dpm).\",\n \"The second standard contained an amount of carbon-14 in scintillant that produces 100,600 dpm.\",\n \"Images were taken using 1.1 pixel binning with background subtraction, and values of counts per second (cps) were acquired.\",\n \"The cps then was multiplied by 60 to obtain counts per minute (cpm).\",\n \"Each discharge is representative of the energy released by a disintegrating beta particle in the form of a photon.\",\n \"“Dpm” divided by “cpm” then provided an approximate efficiency for the camera, which was used for all data values to estimate actual photons emitted.\",\n \"For example, where the carbon-14 standard=3.6 cps and scintillant alone=2.2 cps, the difference=1.4 cps (actual cps)=84 cpm; and 100,600 dpm divided by 84 cpm=1,198 discharges/count (photons/count), which is used as a conversion factor to calculate the number of photons emitted by plant and detected by the camera Using the above conditions and calculation, various wild type and transformed plants were examined and photons/mm2/sec were determined.\",\n \"No visibly detectable bioluminescence was observed in the Columbia wt plants, the 35S LUC-transformed plants or Supernova plants, whereas all of the TK1829 transformed plants were visibly bioluminescent.\",\n \"As shown in Table 4, a threshold value of visible bioluminescence occurs between about 700,000 photons/mm2/sec and 1×106 photons/mm2/sec (compare Tables 4c and 4d; corresponding to 126.4 and 1562.4 pg luc+/μg protein, respectively, as determined by luminometry and spectrophotometry; see Example 2B).\",\n \"TABLE 4 Photon Emission Plant Comparison (cts/mm2/sec) a) Columbia wild-type 16.8 b) 35S:Luc 10,063.0 c) Supernova 701,669.0 d) Tk1829-19-transformed Columbia 1,061,668.0 e) TK1829-11-transformed Supernova 7,160,446.0 f) TK1829-11-transformed Supernova 52,604,180.0 with Norflurazon These results demonstrate that the amount of light produced by the various transformed plants correlates with the amount of luciferase expressed in the plants, and establish standard conditions defining a threshold level of light emission required for visible bioluminescence.\",\n \"These results further provide guidelines for generating other constructs, which contain various combinations of regulatory elements, that can be examined for the ability to confer visible bioluminescence on a plant.\",\n \"For example, by comparing a plant transformed with a recombinant nucleic acid molecule comprising a nucleotide sequence encoding a bioluminescent polypeptide operatively linked to one or more regulatory elements, with plants transformed with a 35S:Luc construct or a cab2:Luc construct, recombinant nucleic acid molecules encompassed within the present invention and useful for generating visibly bioluminescent plant cells or transgenic plants comprising such cells can be obtained.\",\n \"More directly, by transforming plants with various constructs and determining photon emission under standardized conditions as described above, visibly bioluminescent plants can be obtained.\",\n \"Furthermore, the results disclosed herein indicate that plant cells other than Arabidopsis plant cells can be genetically modified using, for example, a TK1829 recombinant nucleic acid molecule, thus providing a means to obtain different species, strains and varieties of visibly bioluminescent transgenic plants.\",\n \"In this respect, genetically modified petunia plants that exhibit exceptional bioluminescence also have been generated using the disclosed compositions and methods.\",\n \"D. Preparation and Characterization of Bioluminescent Petunia Plants pMT-OM-LUC+ or pACT-OM-LUC+ expression constructs were introduced from MT-OM-LUC+ or pACT-OM-LUC+ into Agrobacterium strain ABI 1 and in planta transformation of Arabidopsis and of petunia plants were performed as described above.\",\n \"Whole leaves from 10 week old petunia plants were removed and washed in warm soapy water.\",\n \"Leaves were sterilized in 10% (v/v)chlorine for 15 min and rinsed 4 times with sterile water.\",\n \"Squares of approximately 1 cm were cut from the leaves under sterile conditions with an scalpel and cultured for one day in pre-culture medium (MS medium supplemented with 1 mg/l of 6-benzylaminopurine (BAP) and 0.1 μg/ml of alpha-naphthalene acetic acid (NAA).\",\n \"A. tumefaciens strain ABI carrying one of the plasmids was used to inoculate the pieces of leaves.\",\n \"Bacteria from a single colony were grown at 28° C. overnight in liquid LB medium on a rotary shaker (250 rpm).\",\n \"The medium was supplemented with 50 μg/ml kanamycin, 100 μg/ml spectinomycin and 34 μg/ml chloramphenicol.\",\n \"The next day, a 1:10 dilution was made into LB medium (without antibiotics) and sub-cultured cells were grown for 4 to 5 hr.\",\n \"Cultures were diluted to an OD (660 nm) of 0.05 to 0.07 with pre-culture liquid medium.\",\n \"Explants were inoculated with a log phase solution of Agrobacterium adjusted to approximately 5×108 cells.\",\n \"The leaf pieces were allow to soak for 5 min in the bacteria solution, blotted on sterile Whatman filter discs to remove excess of solution, then replaced in the original pre-culture plates and incubated for 2 days at 23° C. under low light.\",\n \"The leaf squares were transferred into shoot proliferation medium (pre-culture medium containing 75 μg/ml gentamycin and 500 μg/ml carbenicillin).\",\n \"After 4 weeks, shoots were excised and placed on rooting medium (MS medium supplemented with 60 μg/ml gentamycin and 500 μg/ml carbenicillin).\",\n \"After approximately 5 weeks, the plants were transferred to soil.\",\n \"As shown in Table 5, Arabidopsis plants containing the MT-OM-LUC+ construct (designated “31” in Table 5) or the ACT-OM-LUC+ construct (“#27” and “#17”) were visibly biolunminescent (i.e., greater than 750,000 photons of light/mm2/sec (column labeled “photons”).\",\n \"These results demonstrate that the methods of the invention can be used to make various constructs, which, when introduced into plant cells, result in the generation of visibly bioluminescent plants.\",\n \"Similarly, introduction of the TK 1829 construct or the ACT-OM-LUC+ construct into petunia plants resulted in the generation of bioluminescent petunia plants.\",\n \"Although petunia plants containing the Mr-OM-LUC+ construct were not examined for photon counts, the mean RLU/μg was similar to those for the TK 1828 and ACT-OM-LUC+ transformed petunias and the plants were visibly bioluminescent to the eye.\",\n \"These results demonstrate that the constructs of the invention are useful for generating a variety of visibly bioluminescent plants.\",\n \"Together, the results of these studies demonstrate the broad utility of the claimed methods for generating visibly bioluminescent plants.\",\n \"Although the invention has been described with reference to the above examples, it will be understood that modifications and variations are encompassed within the spirit and scope of the invention.\",\n \"Accordingly, the invention is limited only by the following claims.\",\n \"TABLE 5 21 day old Weight OD OD conc.\",\n \"mean pgLuc/ plants ug (750 nm) (750 nm) (ug/ul) RLU 1 RLU 2 RLU/ug ug cts/mm2/sec photons MT Line 31902 31 99.3 0.086 0.086 0.0238844 128.1 132.8 1092.34395 15.24 2408.4 2,885,263.20 Actin Lines Actin 2-7 Col 31902 #27 98.9 0.047 0.046 0.0111278 38.37 39.6 700.68 9.80 911.64 1,092,144.72 Actin 1-1 Col 31902 #17 98.2 0.109 0.109 0.0313123 318.4 331.5 2,075.54 28.81 3342.4 4,004,195.20 Petunia Actin Line T3 102 0.062 0.065 5.0861595 31.46 30.96 1.23 0.02 840 1,006,320.00 TK1829 Line T5 96 0.105 101 84.933336 525.9 520.1 1.23 0.02 3385.7 4,056,068.60 MT Line T3 90 0.106 0.105 3.6981111 22.82 20.23 1.16 0.02 n/a control 27 0.089 0.09 0.0150032 0.028 0.021 0.33 0.00 16.35 19,587.30\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473945"}}},{"rowIdx":770,"cells":{"text":{"kind":"list like","value":[["Facilitating legal interception of ip connections","A method of facilitating the legal interception of IP connections, where two or more terminals can communicate with each other over the Internet using IPSec to provide security.","The method comprises allocating to each terminal T1,T2 a public/private key pair for use in negotiating IKE and IPSec Security Associations (SAs) with other terminals.","Where a terminal T1,T2 is coupled to the Internet via an access network 1,2, the private key of that terminal is stored within the access network at an interception server S1,S2.","When an IP connection is initiated to or from a terminal T1,T2 on which a legal interception order has been placed, the private key stored for that terminal T1,T2 within the access network 1,2 is used to intercept the connection."],["1.A method of facilitating the legal interception of network connections, where two or more terminals can communicate with each other over insecure networks using a standard security protocol to provide a secure session, the method comprising: allocating to each terminal at least one public/private key pair for use in negotiating session encryption keys with other terminals; where a terminal is coupled to an interconnecting network via an access network, storing the private key of that terminal within the access network; and when a connection is initiated to or from a terminal on which a legal interception order has been placed, using the private key stored for that terminal within the access network to intercept the communication, wherein the access network for a terminal interposes itself between that terminal and a remote node during the negotiation of session encryption keys between the terminal and the remote node, such that the access network has a knowledge of negotiated session encryption keys.","2.The method of claim 1, wherein said access network is one of a wireless telecommunication network, a fixed line telephone network, and a cable network.","3.The method of claim 1, where the access network is a wireless telecommunication networks, and the private keys for subscribers are stored at a network switch or at a server coupled to a switch.","4.The method of claim 1, wherein said insecure network is the Internet.","5.The method of claim 4, wherein said secure session is established using IPSec.","6.The method of claim 5, wherein said public/private key pair is used to negotiate IKE and IPSec Security Associations (SAs) comprising said session encryption keys.","7.The method of claim 6, wherein, following the receipt of a request for an ISAKMP SA at the access network of a terminal initiating an IP connection, the access network negotiates an ISAKMP SA with a remote node on behalf of the initiating terminal, and passes the SA parameters to the initiating terminal over a secure connection.","8.The method of claim 7, wherein, once the ISAKMP SA has been established, if the initiating terminal sends a request for the establishment of IPSec SAs towards a destination terminal, the access network determines whether or not a legal interception order has been placed on that terminal or on the terminal to which the initiating terminal wishes to connect and, if such an order has been placed, the access network interposes itself between the terminal and the remote node during the negotiation of a pair of IPSec SAs.","9.The method of claim 6 and comprising: receiving at an access network, the “first” network, a request for establishment of an ISAKMP SA from an initiating terminal; forwarding the request to the access network, the “second” network, of the other terminal; making a decision at the second network on legal interception; and if legal interception is required, negotiating an ISAKMP SA directly between the second network and the initiating terminal and negotiating a second ISAKMP SA directly between the first access network and the destination terminal.","10.The method of claim 9, wherein when the first access network receives from the initiating terminal a request for the establishment of IPSec SAs, the second access network negotiates a pair of IPSec SAs directly with the initiating terminal whilst the first access node negotiates a second pair of IPSec SAs directly with the destination terminal.","11.The method of claim 1 further comprising passing a private key of a terminal to be monitored from one access network to another so that one access network is in possession of the private keys of both or all parties involved in an IP connection.","12.The method of claim 11, wherein said private key is passed from one access network to another without explicitly disclosing the private key to the receiving access network.","13.The method of claim 12, wherein said private key is passed from one access network to another as part of an executable code which can be executed by the receiving network to facilitate negotiation of session encryption keys with a terminal using the receiving network as access network.","14.The method of claim 1 further comprising storing the public/private key pair allocated to a terminal in a memory of or coupled to the terminal in such a way that the user of the terminal cannot alter the private key without the consent of the operator of the relevant access network.","15.The method of claim 1, wherein a node within the access network for a terminal negotiates session encryption keys for both communication directions, on behalf of that terminal, with a remote terminal or a node within the access network for that remote terminal.","16.The method of claim 1, wherein nodes within the access networks of a pair of terminals negotiate session encryption keys with the respective remote terminals, and during interception of a connection each node intercepts communications in one direction, with at least one of the nodes echoing intercepted communications to the other of the nodes.","17.The method of claim 1 further comprising storing in the access network terminal security capabilities.","18.A server for use in intercepting IP connections between two or more terminals, the server comprising: a memory for storing the private keys of public/private key pairs of respective terminals, a first processing means for identifying when legal interception is to be carried out on a connection to or from a terminal, a second processing means for interposing the server between that terminal, and a communication means for receiving communications from a remote node during the negotiation of session encryption keys between the terminal and the remote node, such that the access network has a knowledge of negotiated session encryption keys, and a third processing means for intercepting the connection using the private key of the terminal.","19.A system for facilitating the legal interception of network connections, the system comprising: at least two terminals adapted to communicate with each other over insecure networks using a standard security protocol to provide a secure session, wherein each terminal has instructions for: a means for allocating a least one public/private key pair for use in negotiating session encryption keys with other terminals; a means for storing the private key of that terminal within the access network when a terminal is coupled to an interconnecting network via an access network; and a means for using the private key stored for that terminal within the access network to intercept the communication when a connection is initiated to or from a terminal on which a legal interception order has been placed, and a means for interposing between that terminal and a remote node during the negotiation of session encryption keys between the terminal and the remote node, such that the access network has a knowledge of negotiated session encryption keys."],["<SOH> BACKGROUND TO THE INVENTION <EOH>It is now possible to establish various forms of connection over the internet including data connections as well as voice and video telephony connections.","As the speed and extent of the Internet increase, the use of voice and video telephony can be expected to grow.","Whilst current technology tends to restrict IP telephony to communications between computer terminals coupled to the Internet, tomorrow's technology will provide for IP telephony between small dedicated telephony terminals, and other mobile devices such as PDAs, palmtop computers etc.","In order to allow such devices to gain widespread acceptance, a key issue which must be addressed is that of security.","The two main security concerns are the avoidance of unauthorised eavesdropping, and the need to authenticate terminals involved in a communication (i.e.","to ensure that the terminal which a “subscriber” connects to is the terminal which the subscriber intends to connect to and vice versa).","However, these concerns are not unique to IP telephony, and are common to many different forms of IP communication.","An Internet Engineering Task Force (IETF) standard known as IPsec (RFC2401) has been defined and provides for the creation of a secure connection between parties using IPv4 and IPv6.In the IPsec model the end points of the secure connection are identified by their IP addresses.","In order to allow IPSec packets to be properly encapsulated and decapsulated it is necessary to associate security services and a key between the traffic being transmitted and the remote node which is the intended recipient of the traffic.","The construct used for this purpose is a “Security Association” (SA).","SAs are negotiated between peer nodes using a mechanism known as “Internet Key Exchange” (IKE), and are allocated an identification known as a “Security Parameter Index” (SPI).","The appropriate SA is identified to the receiving node by including the corresponding SPI in the headers of the transmitted data packets.","Details of the existing SAs and the respective SPIs are maintained in a Security Association Database (SAD) which is associated with each IPSec node.","As already noted, IPSec SAs are negotiated using the IKE mechanism.","More particularly, IPSec SAs make use of IKE phase 2 .","IKE phase 1 involves the negotiation of an IKE SA.","FIG.","1 illustrates in general terms the two phases of IE.","When IKE phase 1 is initiated between two nodes, communications are carried out in the open.","The mechanisms used must therefore be extremely secure and inevitably computationally intensive.","At the end of phase 1 both nodes are authenticated to each other, and a shared secret is established between them.","IKE phase 2 makes use of the IKE SA to negotiate one or more IPSec SAs.","As the phase 2 negotiations are carried out within a secure “shell”, they can be much less computationally intensive than the phase 1 negotiation.","Whilst a new IKE SA may be negotiated only infrequently (e.g.","one a day or once a week), IPSec SAs may be negotiated every few minutes.","IKE phases 1 and 2 are illustrated in more detail in FIGS.","2 and 3 .","IPSec makes use of one or both of the Authentication Header (AH) and Encapsulation Security Payload (ESP) protocols which in turn make use of the corresponding established IPSec SA.","Both of these protocols provide for the authentication of sent data packets whilst ESP provides in addition for the encryption of user data.","The use of AH and/or ESP is agreed upon by the communicating nodes during the IKE negotiations.","It is expected ESP will be preferred for VoIP due to its use of encryption.","The precise way in which IPSec is implemented in a system depends to a large extent upon the security policy of the organisation wishing to employ IPSec.","For example, the organisation may specify end-points (e.g.","user terminals) to which IP packets may be sent, or from which they may be received, the particular security levels to be used for encrypting packets, etc.","Policy is stored in a Security Policy Database (SPD) which is also associated with each IPSec node.","Typically, the SPD is distributed amongst a plurality of entities of the IPSec node."],["<SOH> SUMMARY OF THE INVENTION <EOH>Traditional circuit switched telephone networks make provision for the legal interception of telephone calls.","Such interception must be instigated by the appropriate authorities and is an important weapon against crime.","Understandably, it is desirable to make provision for the legal interception of IP connections (whether data, VoIP, video, etc).","However, this presents a problem as the IP security protocol which will become the de facto standard is IPSec, and IPSec has been designed to provide terminal-to-terminal security involving strong encryption.","It is an object of the present invention to overcome the problems inherent to IPSec, and in particular to facilitate the legal interception of IP connections.","According to a first aspect of the present invention there is provided a method of facilitating the legal interception of IP connections, where two or more terminals can communicate with each other over the Internet using IPSec to provide security, the method comprising: allocating to each terminal a public/private key pair for use in negotiating IKE and IPSec Security Associations (SAs) with other terminals; where a terminal is coupled to the Internet via an access network, storing the private key of that terminal within the access network; and when an IP connection is initiated to or from a terminal on which a legal interception order has been placed, using the private key stored for that terminal within the access network to intercept the connection.","The present invention is applicable in particular to facilitating the interception of IP connections made to or from terminals coupled to telecommunication networks, e.g.","wireless telecommunication networks, fixed line telephone networks, and cable networks.","Such networks present the authorities with confidential access points to the connections to be intercepted.","Where the access network is a wireless telecommunication networks, the private keys for subscribers may be stored at an MSC (GSM network) or at an RNC (UMS/3GPP network) or their equivalents, or at a server coupled to the MSC/RNC.","Where the access network is a fixed line telephone network, the private keys may be stored at the local exchanges of subscribers, or at servers coupled to the local exchanges.","The invention is applicable to IP connections between two or more terminals each of which is coupled to the Internet via an access network holding the necessary private keys.","The invention is also applicable to IP connections extending between one of these such terminals and another terminal which handles ISAK negotiations itself, without the aid of an access network holding the private key of the terminal.","In certain embodiments of the invention, following the receipt of a request for an IKE phase 1 SA at the access network of a terminal initiating an IP connection, the access network negotiates an ISAKMP SA with a remote node on behalf of the initiating terminal, and passes the SA parameters (including the established shared key) to the initiating terminal over a secure connection.","The remote node may be the terminal with which the initiating terminal wishes to establish a connection, or may be a node to which that terminal is coupled.","Once the ISAKMP SA has been established, if the initiating terminal sends a request for the establishment of an IKE phase 2 SA intended for the destination terminal, the access network determines whether or not a legal interception order has been placed on that terminal or on the terminal to which the initiating terminal wishes to connect.","If such an order has been placed, the access network interposes itself between the initiating terminal and the remote node during the negotiation of a pair of IKE phase 2 SAs.","An important consideration is that neither the initiating terminal, nor the terminal to which the initiating terminal connects, must know that an interception is being made.","In other embodiments of the invention, and in which both of the terminals between which it is desired to establish a connection are coupled to the Internet via the same or different access networks, receipt at an access network (the “first” network) of a request for establishment of an IKE phase 1 from an initiating terminal causes that first network to forward the request to the access network (the “second” network) of the other terminal.","The second network then makes a decision on legal interception.","If legal interception is required, the second network negotiates an ISAP SA directly with the initiating terminal.","The first network negotiates a second ISAKMP SA directly with the destination terminal.","When the first access network receives from the initiating terminal a request for the establishment of IPSec SAs, the second access network negotiates a pair of SAs directly with the initiating terminal whilst the first access node negotiates a second pair of SAs directly with the destination terminal.","In this way, each of access networks has access to one of the two communication channels.","One of the access networks may relay the channel which it is monitoring to the other access network (or to some other node).","In still other embodiments of the present invention, a private key of a terminal to be monitored may be passed from one access network to another so that one access network is in possession of the private keys of both (or all) parties involved in an IP connection.","This may be achieved in such a way that the private key is not explicitly disclosed to the receiving access network (e.g.","by embedding the key in executable code).","Preferably, IKE Phase 1 (and Phase 2 if appropriate) negotiations handled by an access network are handled by a server.","This server may be coupled to or incorporated into a switch of the access network.","Preferably, the public/private key pair allocated to a terminal is stored in a memory of or coupled to the terminal in such a way that the user of the terminal cannot alter the private key without the consent of the operator of the relevant access network.","The invention is applicable in particular to facilitating the interception of data calls although it is also applicable to VoIP calls and other forms of IP telephony including videoconferencing.","According to a second aspect of the present invention there is provided a server for use in intercepting IP connections between two or more terminals, the server comprising a memory for storing the private keys of public/private key pairs of respective terminals, and processing means for identifying when legal interception is to be carried out on a connection to or from a terminal, and for intercepting that connection using the private key of the terminal."],["FIELD OF THE INVENTION The present invention relates to a method and apparatus for facilitating legal interception of IP connections.","BACKGROUND TO THE INVENTION It is now possible to establish various forms of connection over the internet including data connections as well as voice and video telephony connections.","As the speed and extent of the Internet increase, the use of voice and video telephony can be expected to grow.","Whilst current technology tends to restrict IP telephony to communications between computer terminals coupled to the Internet, tomorrow's technology will provide for IP telephony between small dedicated telephony terminals, and other mobile devices such as PDAs, palmtop computers etc.","In order to allow such devices to gain widespread acceptance, a key issue which must be addressed is that of security.","The two main security concerns are the avoidance of unauthorised eavesdropping, and the need to authenticate terminals involved in a communication (i.e.","to ensure that the terminal which a “subscriber” connects to is the terminal which the subscriber intends to connect to and vice versa).","However, these concerns are not unique to IP telephony, and are common to many different forms of IP communication.","An Internet Engineering Task Force (IETF) standard known as IPsec (RFC2401) has been defined and provides for the creation of a secure connection between parties using IPv4 and IPv6.In the IPsec model the end points of the secure connection are identified by their IP addresses.","In order to allow IPSec packets to be properly encapsulated and decapsulated it is necessary to associate security services and a key between the traffic being transmitted and the remote node which is the intended recipient of the traffic.","The construct used for this purpose is a “Security Association” (SA).","SAs are negotiated between peer nodes using a mechanism known as “Internet Key Exchange” (IKE), and are allocated an identification known as a “Security Parameter Index” (SPI).","The appropriate SA is identified to the receiving node by including the corresponding SPI in the headers of the transmitted data packets.","Details of the existing SAs and the respective SPIs are maintained in a Security Association Database (SAD) which is associated with each IPSec node.","As already noted, IPSec SAs are negotiated using the IKE mechanism.","More particularly, IPSec SAs make use of IKE phase 2.IKE phase 1 involves the negotiation of an IKE SA.","FIG.","1 illustrates in general terms the two phases of IE.","When IKE phase 1 is initiated between two nodes, communications are carried out in the open.","The mechanisms used must therefore be extremely secure and inevitably computationally intensive.","At the end of phase 1 both nodes are authenticated to each other, and a shared secret is established between them.","IKE phase 2 makes use of the IKE SA to negotiate one or more IPSec SAs.","As the phase 2 negotiations are carried out within a secure “shell”, they can be much less computationally intensive than the phase 1 negotiation.","Whilst a new IKE SA may be negotiated only infrequently (e.g.","one a day or once a week), IPSec SAs may be negotiated every few minutes.","IKE phases 1 and 2 are illustrated in more detail in FIGS.","2 and 3.IPSec makes use of one or both of the Authentication Header (AH) and Encapsulation Security Payload (ESP) protocols which in turn make use of the corresponding established IPSec SA.","Both of these protocols provide for the authentication of sent data packets whilst ESP provides in addition for the encryption of user data.","The use of AH and/or ESP is agreed upon by the communicating nodes during the IKE negotiations.","It is expected ESP will be preferred for VoIP due to its use of encryption.","The precise way in which IPSec is implemented in a system depends to a large extent upon the security policy of the organisation wishing to employ IPSec.","For example, the organisation may specify end-points (e.g.","user terminals) to which IP packets may be sent, or from which they may be received, the particular security levels to be used for encrypting packets, etc.","Policy is stored in a Security Policy Database (SPD) which is also associated with each IPSec node.","Typically, the SPD is distributed amongst a plurality of entities of the IPSec node.","SUMMARY OF THE INVENTION Traditional circuit switched telephone networks make provision for the legal interception of telephone calls.","Such interception must be instigated by the appropriate authorities and is an important weapon against crime.","Understandably, it is desirable to make provision for the legal interception of IP connections (whether data, VoIP, video, etc).","However, this presents a problem as the IP security protocol which will become the de facto standard is IPSec, and IPSec has been designed to provide terminal-to-terminal security involving strong encryption.","It is an object of the present invention to overcome the problems inherent to IPSec, and in particular to facilitate the legal interception of IP connections.","According to a first aspect of the present invention there is provided a method of facilitating the legal interception of IP connections, where two or more terminals can communicate with each other over the Internet using IPSec to provide security, the method comprising: allocating to each terminal a public/private key pair for use in negotiating IKE and IPSec Security Associations (SAs) with other terminals; where a terminal is coupled to the Internet via an access network, storing the private key of that terminal within the access network; and when an IP connection is initiated to or from a terminal on which a legal interception order has been placed, using the private key stored for that terminal within the access network to intercept the connection.","The present invention is applicable in particular to facilitating the interception of IP connections made to or from terminals coupled to telecommunication networks, e.g.","wireless telecommunication networks, fixed line telephone networks, and cable networks.","Such networks present the authorities with confidential access points to the connections to be intercepted.","Where the access network is a wireless telecommunication networks, the private keys for subscribers may be stored at an MSC (GSM network) or at an RNC (UMS/3GPP network) or their equivalents, or at a server coupled to the MSC/RNC.","Where the access network is a fixed line telephone network, the private keys may be stored at the local exchanges of subscribers, or at servers coupled to the local exchanges.","The invention is applicable to IP connections between two or more terminals each of which is coupled to the Internet via an access network holding the necessary private keys.","The invention is also applicable to IP connections extending between one of these such terminals and another terminal which handles ISAK negotiations itself, without the aid of an access network holding the private key of the terminal.","In certain embodiments of the invention, following the receipt of a request for an IKE phase 1 SA at the access network of a terminal initiating an IP connection, the access network negotiates an ISAKMP SA with a remote node on behalf of the initiating terminal, and passes the SA parameters (including the established shared key) to the initiating terminal over a secure connection.","The remote node may be the terminal with which the initiating terminal wishes to establish a connection, or may be a node to which that terminal is coupled.","Once the ISAKMP SA has been established, if the initiating terminal sends a request for the establishment of an IKE phase 2 SA intended for the destination terminal, the access network determines whether or not a legal interception order has been placed on that terminal or on the terminal to which the initiating terminal wishes to connect.","If such an order has been placed, the access network interposes itself between the initiating terminal and the remote node during the negotiation of a pair of IKE phase 2 SAs.","An important consideration is that neither the initiating terminal, nor the terminal to which the initiating terminal connects, must know that an interception is being made.","In other embodiments of the invention, and in which both of the terminals between which it is desired to establish a connection are coupled to the Internet via the same or different access networks, receipt at an access network (the “first” network) of a request for establishment of an IKE phase 1 from an initiating terminal causes that first network to forward the request to the access network (the “second” network) of the other terminal.","The second network then makes a decision on legal interception.","If legal interception is required, the second network negotiates an ISAP SA directly with the initiating terminal.","The first network negotiates a second ISAKMP SA directly with the destination terminal.","When the first access network receives from the initiating terminal a request for the establishment of IPSec SAs, the second access network negotiates a pair of SAs directly with the initiating terminal whilst the first access node negotiates a second pair of SAs directly with the destination terminal.","In this way, each of access networks has access to one of the two communication channels.","One of the access networks may relay the channel which it is monitoring to the other access network (or to some other node).","In still other embodiments of the present invention, a private key of a terminal to be monitored may be passed from one access network to another so that one access network is in possession of the private keys of both (or all) parties involved in an IP connection.","This may be achieved in such a way that the private key is not explicitly disclosed to the receiving access network (e.g.","by embedding the key in executable code).","Preferably, IKE Phase 1 (and Phase 2 if appropriate) negotiations handled by an access network are handled by a server.","This server may be coupled to or incorporated into a switch of the access network.","Preferably, the public/private key pair allocated to a terminal is stored in a memory of or coupled to the terminal in such a way that the user of the terminal cannot alter the private key without the consent of the operator of the relevant access network.","The invention is applicable in particular to facilitating the interception of data calls although it is also applicable to VoIP calls and other forms of IP telephony including videoconferencing.","According to a second aspect of the present invention there is provided a server for use in intercepting IP connections between two or more terminals, the server comprising a memory for storing the private keys of public/private key pairs of respective terminals, and processing means for identifying when legal interception is to be carried out on a connection to or from a terminal, and for intercepting that connection using the private key of the terminal.","BRIEF DESCRIPTION OF THE DRAWINGS FIG.","1 illustrates at a general level the signalling between two nodes during a secure data connection establishment process; FIG.","2 illustrates at a more detailed level the signalling involved in an IKE phase 1 of the process of FIG.","1; FIG.","3 illustrates a Quick Mode message exchange of an IKE phase 2 of the process of FIG.","1; FIG.","4 illustrates schematically a system for providing IP telephony between user terminals; FIG.","5 is a signalling diagram illustrating a first method of facilitating legal interception of an IP connection between two of the terminals in FIG.","4; and FIG.","6 is a signalling diagram illustrating a second method of facilitating legal interception of an IP connection between two of the terminals in FIG.","4.DETAILED DESCRIPTION OF A PREFERRED EMBODIMENT The method which will now be described makes use of features described in the following documents: [IPsec] RFC 2401, Security Architecture for the Internet Protocol, November 1998; [IKE] RFC 2409, The Internet Key Exchange (IKE), November 1998; [ISAKMP] RFC 2408, Internet Security Association and Key Management Protocol, November 1998; IETG Draft “Running DIE Phase 2 over Artificial Kerberos IKE SA”, draft-ietf-kink-ike-over-kklmp-00.txt; RFC 2459, Internet X.509 Public Key Inastmeture Certificate and CRL Profile.","Reference should be made to these documents for a fuller understanding of the method.","FIG.","4 illustrates a scenario in which a pair of mobile wireless terminals T1,T2 are coupled to the Internet via respective access networks 1,2.For the purpose of this example, the access networks 1,2 are considered to be mobile telecommunication networks, with the wireless terminals T1,T2 being mobile telephones (or other mobile wireless terminals having mobile telephone functionality).","It is noted that whilst current digital networks (such as GSM, PDS, and digital AMP networks) allow user terminals to access the Internet, Internet access will be greatly enhanced by the introduction of data services (such as GPRS) and as third future generation mobile networks (such a UMTS) are introduced.","The terminals T1,T2 are assumed to be specifically designed to facilitate IP data calls (and possibly IP telephony applications including VoIP and video telephony).","Illustrated in FIG.","4 are a pair of interception servers S1,S2 associated with respective access networks 1,2.Each server S1,S2 is coupled to that “switch” (e.g.","MSC or RNC—not shown in FIG.","4) of the associated access network 1,2 which handles IP calls made to or from terminals connected to the network.","As explained above, in order to make use of IPSec to secure IP connections, a terminal should be in possession of a public/private key pair (Ppriv/Ppubl).","This key pair may be stored in a fixed memory of the terminal or may be stored in the terminal's SIM card.","It is assumed to have been stored there by the operator of the access network to which the terminal's owner subscribes, prior to the terminal being presented to the owner.","Once stored in the terminal (or SIM card) it is not possible for the owner to change the key pair without the consent of the network operator.","The operator retains a copy of the subscriber's public/private key pair and stores this in a look-up table of the interception server S1,S2.The server also has a knowledge of the terminal's capabilities including the available cryptographic algorithms and the default security policy.","The server maintains a record of those subscribers on whom legal interception orders have been placed by the authorities.","When a terminal (say T1 in FIG.","4) wishes to establish an IP connection to another terminal it must first obtain the IP address of the other terminal T2.If the initiating terminal T1 does not know this IP address from the outset, it must obtain it, e.g.","by sending a user entered URL to a Domain Name Server (ENS) which can resolve the address and return it to the initiating terminal.","A already outlined above, in order to secure the connection, the initiating terminal T1 must then negotiate a pair of IPSec SAs with the other (or “destination”) terminal T2.FIG.","5 illustrates the exchange of signalling messages required to establish a pair of SAs.","Firstly, the initiating terminal T1 sends a Request ISAKMP SA message to its access network 1.This message contains the identity of the initiating and destination terminals (which may be the terminals' IMSI codes or telephone numbers).","The request is handled by the access network's interception server S1.The server checks whether or not it holds the private key for initiating terminal T1.If not, the call connection attempt is rejected.","If the server does hold the appropriate private key, the server S1 handles the negotiation of the ISAKMP SA (IKE phase 1) on behalf of the terminal T1 (the terminal T1 does not require IKE Phase 1 capabilities).","In the same way, the interception server 52 in the access network 2 of the “destination” terminal T2 handles the ISAKMP SA negotiation on behalf of that terminal T2.This is possible because the interception servers S1,S2 know the private keys of the respective terminals T1,T2.Upon completion of the negotiation, the relevant ISAKMP SA parameters are returned by the interception servers to their respective terminals.","As the access networks are private networks, typically using security protocols to secure transmission over the radio interface, there is no danger of the returned parameters falling into the wrong hands.","(As an alternative to IKE Phase 1, Kerberos authentication may be used.)","At this stage, the interception servers S1,S2 know the parameters which will be used to secure the subsequent negotiation of IPSec SAs between the terminals (i.e.","IKE Phase 2).","When terminal T1 wishes to begin a secure communication with terminal T2, it sends a Begin IPSec SA negotiation request to terminal T2.This message is intercepted by the access network 1 and the interception server S1 is notified.","The request includes in its header portion the relevant ISAKMP SA data.","The server S1 then checks its records to see if either the initiating or destination terminals have legal interception orders placed on them.","If they do not, the IPSec SA negotiation continues without the involvement of interception server S1.It is noted that once the IPSec SAs have been negotiated in this way, it is not possible for the access network to intercept (and decrypt) traffic carried using the LPSec SAs.","If on the other hand the interception server S1 determines that a legal interception order has been placed on one of the terminals T1 or T2, the server S1 interposes itself between terminals T1 and T2 during the negotiation of the pair of IPSec SAs (similarly server S2 may interpose itself in the IPSec SA negotiation if that server decides that it too must intercept the connection).","As far as terminals T1 and T2 are concerned, they are negotiating the IPSec SAs with each other and are not aware of the role of the server(s) in the process.","Subsequently, encrypted traffic may be intercepted at the server S1, decrypted using the negotiated IPSec SA parameters known to server S1, and routed to an appropriate monitoring point.","An important feature is that neither of the terminals is allowed to run the fill ISAKMP negotiation.","There is illustrated in FIG.","6 signalling associated with an alternative interception scheme.","In this scheme, successful legal interception requires the active participation of both interception servers S1,S2 which again hold the private keys of their respective subscribers, and also assumes that the terminals involved in the connection are capable of handling the IKE phase 1 negotiations (nb.","this latter requirement does not exist for the embodiment of FIG.","5).","When an interception server (S1 in FIG.","6) receives a request for an ISAKMP SA negotiation from an originating terminal T1, that interception server S1 forwards the request to the destination side interception server S2.If the initiating side interception server S1 determines that legal interception is required, a flag is set in the sent request.","When the destination side interception server S2 receives the request it checks the status of the interception flag.","If that flag is set, or if the receiving interception server S2 determines from its own records that legal interception is required, the server S2 performs the first ISAKMP SA negotiation (including the Diffie-Hellman exchange) directly with the initiating terminal T1 on behalf of the destination terminal T2.The destination side interception server S2 also notifies the originating side server S1 that it should negotiate a second ISAKMP SA directly with the destination terminal T2.When the initiating side access network 1 subsequently intercepts an IPSec SA negotiation request sent from the initiating terminal T1 (and intended for the destination terminal T2), the request is forwarded to the destination side interception server S2.The destination side interception server S2 represents itself to terminal T1 as if it were the destination terminal T2 for the purpose of the IPSec phase 2 negotiation.","A pair of IPSec SAs are established between terminal T1 and the interception server S2.At the same time, the destination side interception server S2 instructs the initiating side interception server S1 to negotiate a second pair of IPSec SA with the destination terminal T2.Server S1 therefore represents itself to the destination terminal T2 as if it were terminal T1 during the IPSec phase 2 negotiation.","A second pair of IPSec-SAs are established between server S1 and terminal T2.It is necessary to echo data between the servers S1 and S2 in order to give the servers access to both channels.","Thus, data originating from terminal T1 is reflected by server S2 to server S1.Server S1 then encrypts the data using the appropriate IPSec SA and sends it to terminal T2.Similarly, data originating from terminal T2 is reflected by server S1 to server S2, with server S2 encrypting the data using the appropriate IPSec SA and sending it to terminal T1.It is noted that with the embodiment of FIG.","6 there is no need for “artificial” ISAKMP SA transfers (between servers and terminals).","Rather, the initiating terminals IKE signalling will be processed by the destination side interception server and vice versa.","It will be appreciated by the person of skill in the art that various modifications may be made to the above described embodiments without departing from the scope of the present invention.","For example, in the embodiment described with reference to FIG.","6, the additional delay which results from echoing one monitored channel back from one interception server to the other may be avoided by executing a secure IKE proxy server at the interception server which is monitoring the connection.","This proxy server is sent from the other interception server and contains the private key of the terminal for which it is responsible.","The private key is incorporated into the proxy server in such a way that it is not disclosed to the receiving interception server.","However, when the proxy server is executed, it allows the interception server to conduct the IKE phase 1 and 2 negotiations directly with the initiating and destination terminals.","Thus, the interception server will have access to both traffic channels."]],"string":"[\n [\n \"Facilitating legal interception of ip connections\",\n \"A method of facilitating the legal interception of IP connections, where two or more terminals can communicate with each other over the Internet using IPSec to provide security.\",\n \"The method comprises allocating to each terminal T1,T2 a public/private key pair for use in negotiating IKE and IPSec Security Associations (SAs) with other terminals.\",\n \"Where a terminal T1,T2 is coupled to the Internet via an access network 1,2, the private key of that terminal is stored within the access network at an interception server S1,S2.\",\n \"When an IP connection is initiated to or from a terminal T1,T2 on which a legal interception order has been placed, the private key stored for that terminal T1,T2 within the access network 1,2 is used to intercept the connection.\"\n ],\n [\n \"1.A method of facilitating the legal interception of network connections, where two or more terminals can communicate with each other over insecure networks using a standard security protocol to provide a secure session, the method comprising: allocating to each terminal at least one public/private key pair for use in negotiating session encryption keys with other terminals; where a terminal is coupled to an interconnecting network via an access network, storing the private key of that terminal within the access network; and when a connection is initiated to or from a terminal on which a legal interception order has been placed, using the private key stored for that terminal within the access network to intercept the communication, wherein the access network for a terminal interposes itself between that terminal and a remote node during the negotiation of session encryption keys between the terminal and the remote node, such that the access network has a knowledge of negotiated session encryption keys.\",\n \"2.The method of claim 1, wherein said access network is one of a wireless telecommunication network, a fixed line telephone network, and a cable network.\",\n \"3.The method of claim 1, where the access network is a wireless telecommunication networks, and the private keys for subscribers are stored at a network switch or at a server coupled to a switch.\",\n \"4.The method of claim 1, wherein said insecure network is the Internet.\",\n \"5.The method of claim 4, wherein said secure session is established using IPSec.\",\n \"6.The method of claim 5, wherein said public/private key pair is used to negotiate IKE and IPSec Security Associations (SAs) comprising said session encryption keys.\",\n \"7.The method of claim 6, wherein, following the receipt of a request for an ISAKMP SA at the access network of a terminal initiating an IP connection, the access network negotiates an ISAKMP SA with a remote node on behalf of the initiating terminal, and passes the SA parameters to the initiating terminal over a secure connection.\",\n \"8.The method of claim 7, wherein, once the ISAKMP SA has been established, if the initiating terminal sends a request for the establishment of IPSec SAs towards a destination terminal, the access network determines whether or not a legal interception order has been placed on that terminal or on the terminal to which the initiating terminal wishes to connect and, if such an order has been placed, the access network interposes itself between the terminal and the remote node during the negotiation of a pair of IPSec SAs.\",\n \"9.The method of claim 6 and comprising: receiving at an access network, the “first” network, a request for establishment of an ISAKMP SA from an initiating terminal; forwarding the request to the access network, the “second” network, of the other terminal; making a decision at the second network on legal interception; and if legal interception is required, negotiating an ISAKMP SA directly between the second network and the initiating terminal and negotiating a second ISAKMP SA directly between the first access network and the destination terminal.\",\n \"10.The method of claim 9, wherein when the first access network receives from the initiating terminal a request for the establishment of IPSec SAs, the second access network negotiates a pair of IPSec SAs directly with the initiating terminal whilst the first access node negotiates a second pair of IPSec SAs directly with the destination terminal.\",\n \"11.The method of claim 1 further comprising passing a private key of a terminal to be monitored from one access network to another so that one access network is in possession of the private keys of both or all parties involved in an IP connection.\",\n \"12.The method of claim 11, wherein said private key is passed from one access network to another without explicitly disclosing the private key to the receiving access network.\",\n \"13.The method of claim 12, wherein said private key is passed from one access network to another as part of an executable code which can be executed by the receiving network to facilitate negotiation of session encryption keys with a terminal using the receiving network as access network.\",\n \"14.The method of claim 1 further comprising storing the public/private key pair allocated to a terminal in a memory of or coupled to the terminal in such a way that the user of the terminal cannot alter the private key without the consent of the operator of the relevant access network.\",\n \"15.The method of claim 1, wherein a node within the access network for a terminal negotiates session encryption keys for both communication directions, on behalf of that terminal, with a remote terminal or a node within the access network for that remote terminal.\",\n \"16.The method of claim 1, wherein nodes within the access networks of a pair of terminals negotiate session encryption keys with the respective remote terminals, and during interception of a connection each node intercepts communications in one direction, with at least one of the nodes echoing intercepted communications to the other of the nodes.\",\n \"17.The method of claim 1 further comprising storing in the access network terminal security capabilities.\",\n \"18.A server for use in intercepting IP connections between two or more terminals, the server comprising: a memory for storing the private keys of public/private key pairs of respective terminals, a first processing means for identifying when legal interception is to be carried out on a connection to or from a terminal, a second processing means for interposing the server between that terminal, and a communication means for receiving communications from a remote node during the negotiation of session encryption keys between the terminal and the remote node, such that the access network has a knowledge of negotiated session encryption keys, and a third processing means for intercepting the connection using the private key of the terminal.\",\n \"19.A system for facilitating the legal interception of network connections, the system comprising: at least two terminals adapted to communicate with each other over insecure networks using a standard security protocol to provide a secure session, wherein each terminal has instructions for: a means for allocating a least one public/private key pair for use in negotiating session encryption keys with other terminals; a means for storing the private key of that terminal within the access network when a terminal is coupled to an interconnecting network via an access network; and a means for using the private key stored for that terminal within the access network to intercept the communication when a connection is initiated to or from a terminal on which a legal interception order has been placed, and a means for interposing between that terminal and a remote node during the negotiation of session encryption keys between the terminal and the remote node, such that the access network has a knowledge of negotiated session encryption keys.\"\n ],\n [\n \"<SOH> BACKGROUND TO THE INVENTION <EOH>It is now possible to establish various forms of connection over the internet including data connections as well as voice and video telephony connections.\",\n \"As the speed and extent of the Internet increase, the use of voice and video telephony can be expected to grow.\",\n \"Whilst current technology tends to restrict IP telephony to communications between computer terminals coupled to the Internet, tomorrow's technology will provide for IP telephony between small dedicated telephony terminals, and other mobile devices such as PDAs, palmtop computers etc.\",\n \"In order to allow such devices to gain widespread acceptance, a key issue which must be addressed is that of security.\",\n \"The two main security concerns are the avoidance of unauthorised eavesdropping, and the need to authenticate terminals involved in a communication (i.e.\",\n \"to ensure that the terminal which a “subscriber” connects to is the terminal which the subscriber intends to connect to and vice versa).\",\n \"However, these concerns are not unique to IP telephony, and are common to many different forms of IP communication.\",\n \"An Internet Engineering Task Force (IETF) standard known as IPsec (RFC2401) has been defined and provides for the creation of a secure connection between parties using IPv4 and IPv6.In the IPsec model the end points of the secure connection are identified by their IP addresses.\",\n \"In order to allow IPSec packets to be properly encapsulated and decapsulated it is necessary to associate security services and a key between the traffic being transmitted and the remote node which is the intended recipient of the traffic.\",\n \"The construct used for this purpose is a “Security Association” (SA).\",\n \"SAs are negotiated between peer nodes using a mechanism known as “Internet Key Exchange” (IKE), and are allocated an identification known as a “Security Parameter Index” (SPI).\",\n \"The appropriate SA is identified to the receiving node by including the corresponding SPI in the headers of the transmitted data packets.\",\n \"Details of the existing SAs and the respective SPIs are maintained in a Security Association Database (SAD) which is associated with each IPSec node.\",\n \"As already noted, IPSec SAs are negotiated using the IKE mechanism.\",\n \"More particularly, IPSec SAs make use of IKE phase 2 .\",\n \"IKE phase 1 involves the negotiation of an IKE SA.\",\n \"FIG.\",\n \"1 illustrates in general terms the two phases of IE.\",\n \"When IKE phase 1 is initiated between two nodes, communications are carried out in the open.\",\n \"The mechanisms used must therefore be extremely secure and inevitably computationally intensive.\",\n \"At the end of phase 1 both nodes are authenticated to each other, and a shared secret is established between them.\",\n \"IKE phase 2 makes use of the IKE SA to negotiate one or more IPSec SAs.\",\n \"As the phase 2 negotiations are carried out within a secure “shell”, they can be much less computationally intensive than the phase 1 negotiation.\",\n \"Whilst a new IKE SA may be negotiated only infrequently (e.g.\",\n \"one a day or once a week), IPSec SAs may be negotiated every few minutes.\",\n \"IKE phases 1 and 2 are illustrated in more detail in FIGS.\",\n \"2 and 3 .\",\n \"IPSec makes use of one or both of the Authentication Header (AH) and Encapsulation Security Payload (ESP) protocols which in turn make use of the corresponding established IPSec SA.\",\n \"Both of these protocols provide for the authentication of sent data packets whilst ESP provides in addition for the encryption of user data.\",\n \"The use of AH and/or ESP is agreed upon by the communicating nodes during the IKE negotiations.\",\n \"It is expected ESP will be preferred for VoIP due to its use of encryption.\",\n \"The precise way in which IPSec is implemented in a system depends to a large extent upon the security policy of the organisation wishing to employ IPSec.\",\n \"For example, the organisation may specify end-points (e.g.\",\n \"user terminals) to which IP packets may be sent, or from which they may be received, the particular security levels to be used for encrypting packets, etc.\",\n \"Policy is stored in a Security Policy Database (SPD) which is also associated with each IPSec node.\",\n \"Typically, the SPD is distributed amongst a plurality of entities of the IPSec node.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>Traditional circuit switched telephone networks make provision for the legal interception of telephone calls.\",\n \"Such interception must be instigated by the appropriate authorities and is an important weapon against crime.\",\n \"Understandably, it is desirable to make provision for the legal interception of IP connections (whether data, VoIP, video, etc).\",\n \"However, this presents a problem as the IP security protocol which will become the de facto standard is IPSec, and IPSec has been designed to provide terminal-to-terminal security involving strong encryption.\",\n \"It is an object of the present invention to overcome the problems inherent to IPSec, and in particular to facilitate the legal interception of IP connections.\",\n \"According to a first aspect of the present invention there is provided a method of facilitating the legal interception of IP connections, where two or more terminals can communicate with each other over the Internet using IPSec to provide security, the method comprising: allocating to each terminal a public/private key pair for use in negotiating IKE and IPSec Security Associations (SAs) with other terminals; where a terminal is coupled to the Internet via an access network, storing the private key of that terminal within the access network; and when an IP connection is initiated to or from a terminal on which a legal interception order has been placed, using the private key stored for that terminal within the access network to intercept the connection.\",\n \"The present invention is applicable in particular to facilitating the interception of IP connections made to or from terminals coupled to telecommunication networks, e.g.\",\n \"wireless telecommunication networks, fixed line telephone networks, and cable networks.\",\n \"Such networks present the authorities with confidential access points to the connections to be intercepted.\",\n \"Where the access network is a wireless telecommunication networks, the private keys for subscribers may be stored at an MSC (GSM network) or at an RNC (UMS/3GPP network) or their equivalents, or at a server coupled to the MSC/RNC.\",\n \"Where the access network is a fixed line telephone network, the private keys may be stored at the local exchanges of subscribers, or at servers coupled to the local exchanges.\",\n \"The invention is applicable to IP connections between two or more terminals each of which is coupled to the Internet via an access network holding the necessary private keys.\",\n \"The invention is also applicable to IP connections extending between one of these such terminals and another terminal which handles ISAK negotiations itself, without the aid of an access network holding the private key of the terminal.\",\n \"In certain embodiments of the invention, following the receipt of a request for an IKE phase 1 SA at the access network of a terminal initiating an IP connection, the access network negotiates an ISAKMP SA with a remote node on behalf of the initiating terminal, and passes the SA parameters (including the established shared key) to the initiating terminal over a secure connection.\",\n \"The remote node may be the terminal with which the initiating terminal wishes to establish a connection, or may be a node to which that terminal is coupled.\",\n \"Once the ISAKMP SA has been established, if the initiating terminal sends a request for the establishment of an IKE phase 2 SA intended for the destination terminal, the access network determines whether or not a legal interception order has been placed on that terminal or on the terminal to which the initiating terminal wishes to connect.\",\n \"If such an order has been placed, the access network interposes itself between the initiating terminal and the remote node during the negotiation of a pair of IKE phase 2 SAs.\",\n \"An important consideration is that neither the initiating terminal, nor the terminal to which the initiating terminal connects, must know that an interception is being made.\",\n \"In other embodiments of the invention, and in which both of the terminals between which it is desired to establish a connection are coupled to the Internet via the same or different access networks, receipt at an access network (the “first” network) of a request for establishment of an IKE phase 1 from an initiating terminal causes that first network to forward the request to the access network (the “second” network) of the other terminal.\",\n \"The second network then makes a decision on legal interception.\",\n \"If legal interception is required, the second network negotiates an ISAP SA directly with the initiating terminal.\",\n \"The first network negotiates a second ISAKMP SA directly with the destination terminal.\",\n \"When the first access network receives from the initiating terminal a request for the establishment of IPSec SAs, the second access network negotiates a pair of SAs directly with the initiating terminal whilst the first access node negotiates a second pair of SAs directly with the destination terminal.\",\n \"In this way, each of access networks has access to one of the two communication channels.\",\n \"One of the access networks may relay the channel which it is monitoring to the other access network (or to some other node).\",\n \"In still other embodiments of the present invention, a private key of a terminal to be monitored may be passed from one access network to another so that one access network is in possession of the private keys of both (or all) parties involved in an IP connection.\",\n \"This may be achieved in such a way that the private key is not explicitly disclosed to the receiving access network (e.g.\",\n \"by embedding the key in executable code).\",\n \"Preferably, IKE Phase 1 (and Phase 2 if appropriate) negotiations handled by an access network are handled by a server.\",\n \"This server may be coupled to or incorporated into a switch of the access network.\",\n \"Preferably, the public/private key pair allocated to a terminal is stored in a memory of or coupled to the terminal in such a way that the user of the terminal cannot alter the private key without the consent of the operator of the relevant access network.\",\n \"The invention is applicable in particular to facilitating the interception of data calls although it is also applicable to VoIP calls and other forms of IP telephony including videoconferencing.\",\n \"According to a second aspect of the present invention there is provided a server for use in intercepting IP connections between two or more terminals, the server comprising a memory for storing the private keys of public/private key pairs of respective terminals, and processing means for identifying when legal interception is to be carried out on a connection to or from a terminal, and for intercepting that connection using the private key of the terminal.\"\n ],\n [\n \"FIELD OF THE INVENTION The present invention relates to a method and apparatus for facilitating legal interception of IP connections.\",\n \"BACKGROUND TO THE INVENTION It is now possible to establish various forms of connection over the internet including data connections as well as voice and video telephony connections.\",\n \"As the speed and extent of the Internet increase, the use of voice and video telephony can be expected to grow.\",\n \"Whilst current technology tends to restrict IP telephony to communications between computer terminals coupled to the Internet, tomorrow's technology will provide for IP telephony between small dedicated telephony terminals, and other mobile devices such as PDAs, palmtop computers etc.\",\n \"In order to allow such devices to gain widespread acceptance, a key issue which must be addressed is that of security.\",\n \"The two main security concerns are the avoidance of unauthorised eavesdropping, and the need to authenticate terminals involved in a communication (i.e.\",\n \"to ensure that the terminal which a “subscriber” connects to is the terminal which the subscriber intends to connect to and vice versa).\",\n \"However, these concerns are not unique to IP telephony, and are common to many different forms of IP communication.\",\n \"An Internet Engineering Task Force (IETF) standard known as IPsec (RFC2401) has been defined and provides for the creation of a secure connection between parties using IPv4 and IPv6.In the IPsec model the end points of the secure connection are identified by their IP addresses.\",\n \"In order to allow IPSec packets to be properly encapsulated and decapsulated it is necessary to associate security services and a key between the traffic being transmitted and the remote node which is the intended recipient of the traffic.\",\n \"The construct used for this purpose is a “Security Association” (SA).\",\n \"SAs are negotiated between peer nodes using a mechanism known as “Internet Key Exchange” (IKE), and are allocated an identification known as a “Security Parameter Index” (SPI).\",\n \"The appropriate SA is identified to the receiving node by including the corresponding SPI in the headers of the transmitted data packets.\",\n \"Details of the existing SAs and the respective SPIs are maintained in a Security Association Database (SAD) which is associated with each IPSec node.\",\n \"As already noted, IPSec SAs are negotiated using the IKE mechanism.\",\n \"More particularly, IPSec SAs make use of IKE phase 2.IKE phase 1 involves the negotiation of an IKE SA.\",\n \"FIG.\",\n \"1 illustrates in general terms the two phases of IE.\",\n \"When IKE phase 1 is initiated between two nodes, communications are carried out in the open.\",\n \"The mechanisms used must therefore be extremely secure and inevitably computationally intensive.\",\n \"At the end of phase 1 both nodes are authenticated to each other, and a shared secret is established between them.\",\n \"IKE phase 2 makes use of the IKE SA to negotiate one or more IPSec SAs.\",\n \"As the phase 2 negotiations are carried out within a secure “shell”, they can be much less computationally intensive than the phase 1 negotiation.\",\n \"Whilst a new IKE SA may be negotiated only infrequently (e.g.\",\n \"one a day or once a week), IPSec SAs may be negotiated every few minutes.\",\n \"IKE phases 1 and 2 are illustrated in more detail in FIGS.\",\n \"2 and 3.IPSec makes use of one or both of the Authentication Header (AH) and Encapsulation Security Payload (ESP) protocols which in turn make use of the corresponding established IPSec SA.\",\n \"Both of these protocols provide for the authentication of sent data packets whilst ESP provides in addition for the encryption of user data.\",\n \"The use of AH and/or ESP is agreed upon by the communicating nodes during the IKE negotiations.\",\n \"It is expected ESP will be preferred for VoIP due to its use of encryption.\",\n \"The precise way in which IPSec is implemented in a system depends to a large extent upon the security policy of the organisation wishing to employ IPSec.\",\n \"For example, the organisation may specify end-points (e.g.\",\n \"user terminals) to which IP packets may be sent, or from which they may be received, the particular security levels to be used for encrypting packets, etc.\",\n \"Policy is stored in a Security Policy Database (SPD) which is also associated with each IPSec node.\",\n \"Typically, the SPD is distributed amongst a plurality of entities of the IPSec node.\",\n \"SUMMARY OF THE INVENTION Traditional circuit switched telephone networks make provision for the legal interception of telephone calls.\",\n \"Such interception must be instigated by the appropriate authorities and is an important weapon against crime.\",\n \"Understandably, it is desirable to make provision for the legal interception of IP connections (whether data, VoIP, video, etc).\",\n \"However, this presents a problem as the IP security protocol which will become the de facto standard is IPSec, and IPSec has been designed to provide terminal-to-terminal security involving strong encryption.\",\n \"It is an object of the present invention to overcome the problems inherent to IPSec, and in particular to facilitate the legal interception of IP connections.\",\n \"According to a first aspect of the present invention there is provided a method of facilitating the legal interception of IP connections, where two or more terminals can communicate with each other over the Internet using IPSec to provide security, the method comprising: allocating to each terminal a public/private key pair for use in negotiating IKE and IPSec Security Associations (SAs) with other terminals; where a terminal is coupled to the Internet via an access network, storing the private key of that terminal within the access network; and when an IP connection is initiated to or from a terminal on which a legal interception order has been placed, using the private key stored for that terminal within the access network to intercept the connection.\",\n \"The present invention is applicable in particular to facilitating the interception of IP connections made to or from terminals coupled to telecommunication networks, e.g.\",\n \"wireless telecommunication networks, fixed line telephone networks, and cable networks.\",\n \"Such networks present the authorities with confidential access points to the connections to be intercepted.\",\n \"Where the access network is a wireless telecommunication networks, the private keys for subscribers may be stored at an MSC (GSM network) or at an RNC (UMS/3GPP network) or their equivalents, or at a server coupled to the MSC/RNC.\",\n \"Where the access network is a fixed line telephone network, the private keys may be stored at the local exchanges of subscribers, or at servers coupled to the local exchanges.\",\n \"The invention is applicable to IP connections between two or more terminals each of which is coupled to the Internet via an access network holding the necessary private keys.\",\n \"The invention is also applicable to IP connections extending between one of these such terminals and another terminal which handles ISAK negotiations itself, without the aid of an access network holding the private key of the terminal.\",\n \"In certain embodiments of the invention, following the receipt of a request for an IKE phase 1 SA at the access network of a terminal initiating an IP connection, the access network negotiates an ISAKMP SA with a remote node on behalf of the initiating terminal, and passes the SA parameters (including the established shared key) to the initiating terminal over a secure connection.\",\n \"The remote node may be the terminal with which the initiating terminal wishes to establish a connection, or may be a node to which that terminal is coupled.\",\n \"Once the ISAKMP SA has been established, if the initiating terminal sends a request for the establishment of an IKE phase 2 SA intended for the destination terminal, the access network determines whether or not a legal interception order has been placed on that terminal or on the terminal to which the initiating terminal wishes to connect.\",\n \"If such an order has been placed, the access network interposes itself between the initiating terminal and the remote node during the negotiation of a pair of IKE phase 2 SAs.\",\n \"An important consideration is that neither the initiating terminal, nor the terminal to which the initiating terminal connects, must know that an interception is being made.\",\n \"In other embodiments of the invention, and in which both of the terminals between which it is desired to establish a connection are coupled to the Internet via the same or different access networks, receipt at an access network (the “first” network) of a request for establishment of an IKE phase 1 from an initiating terminal causes that first network to forward the request to the access network (the “second” network) of the other terminal.\",\n \"The second network then makes a decision on legal interception.\",\n \"If legal interception is required, the second network negotiates an ISAP SA directly with the initiating terminal.\",\n \"The first network negotiates a second ISAKMP SA directly with the destination terminal.\",\n \"When the first access network receives from the initiating terminal a request for the establishment of IPSec SAs, the second access network negotiates a pair of SAs directly with the initiating terminal whilst the first access node negotiates a second pair of SAs directly with the destination terminal.\",\n \"In this way, each of access networks has access to one of the two communication channels.\",\n \"One of the access networks may relay the channel which it is monitoring to the other access network (or to some other node).\",\n \"In still other embodiments of the present invention, a private key of a terminal to be monitored may be passed from one access network to another so that one access network is in possession of the private keys of both (or all) parties involved in an IP connection.\",\n \"This may be achieved in such a way that the private key is not explicitly disclosed to the receiving access network (e.g.\",\n \"by embedding the key in executable code).\",\n \"Preferably, IKE Phase 1 (and Phase 2 if appropriate) negotiations handled by an access network are handled by a server.\",\n \"This server may be coupled to or incorporated into a switch of the access network.\",\n \"Preferably, the public/private key pair allocated to a terminal is stored in a memory of or coupled to the terminal in such a way that the user of the terminal cannot alter the private key without the consent of the operator of the relevant access network.\",\n \"The invention is applicable in particular to facilitating the interception of data calls although it is also applicable to VoIP calls and other forms of IP telephony including videoconferencing.\",\n \"According to a second aspect of the present invention there is provided a server for use in intercepting IP connections between two or more terminals, the server comprising a memory for storing the private keys of public/private key pairs of respective terminals, and processing means for identifying when legal interception is to be carried out on a connection to or from a terminal, and for intercepting that connection using the private key of the terminal.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS FIG.\",\n \"1 illustrates at a general level the signalling between two nodes during a secure data connection establishment process; FIG.\",\n \"2 illustrates at a more detailed level the signalling involved in an IKE phase 1 of the process of FIG.\",\n \"1; FIG.\",\n \"3 illustrates a Quick Mode message exchange of an IKE phase 2 of the process of FIG.\",\n \"1; FIG.\",\n \"4 illustrates schematically a system for providing IP telephony between user terminals; FIG.\",\n \"5 is a signalling diagram illustrating a first method of facilitating legal interception of an IP connection between two of the terminals in FIG.\",\n \"4; and FIG.\",\n \"6 is a signalling diagram illustrating a second method of facilitating legal interception of an IP connection between two of the terminals in FIG.\",\n \"4.DETAILED DESCRIPTION OF A PREFERRED EMBODIMENT The method which will now be described makes use of features described in the following documents: [IPsec] RFC 2401, Security Architecture for the Internet Protocol, November 1998; [IKE] RFC 2409, The Internet Key Exchange (IKE), November 1998; [ISAKMP] RFC 2408, Internet Security Association and Key Management Protocol, November 1998; IETG Draft “Running DIE Phase 2 over Artificial Kerberos IKE SA”, draft-ietf-kink-ike-over-kklmp-00.txt; RFC 2459, Internet X.509 Public Key Inastmeture Certificate and CRL Profile.\",\n \"Reference should be made to these documents for a fuller understanding of the method.\",\n \"FIG.\",\n \"4 illustrates a scenario in which a pair of mobile wireless terminals T1,T2 are coupled to the Internet via respective access networks 1,2.For the purpose of this example, the access networks 1,2 are considered to be mobile telecommunication networks, with the wireless terminals T1,T2 being mobile telephones (or other mobile wireless terminals having mobile telephone functionality).\",\n \"It is noted that whilst current digital networks (such as GSM, PDS, and digital AMP networks) allow user terminals to access the Internet, Internet access will be greatly enhanced by the introduction of data services (such as GPRS) and as third future generation mobile networks (such a UMTS) are introduced.\",\n \"The terminals T1,T2 are assumed to be specifically designed to facilitate IP data calls (and possibly IP telephony applications including VoIP and video telephony).\",\n \"Illustrated in FIG.\",\n \"4 are a pair of interception servers S1,S2 associated with respective access networks 1,2.Each server S1,S2 is coupled to that “switch” (e.g.\",\n \"MSC or RNC—not shown in FIG.\",\n \"4) of the associated access network 1,2 which handles IP calls made to or from terminals connected to the network.\",\n \"As explained above, in order to make use of IPSec to secure IP connections, a terminal should be in possession of a public/private key pair (Ppriv/Ppubl).\",\n \"This key pair may be stored in a fixed memory of the terminal or may be stored in the terminal's SIM card.\",\n \"It is assumed to have been stored there by the operator of the access network to which the terminal's owner subscribes, prior to the terminal being presented to the owner.\",\n \"Once stored in the terminal (or SIM card) it is not possible for the owner to change the key pair without the consent of the network operator.\",\n \"The operator retains a copy of the subscriber's public/private key pair and stores this in a look-up table of the interception server S1,S2.The server also has a knowledge of the terminal's capabilities including the available cryptographic algorithms and the default security policy.\",\n \"The server maintains a record of those subscribers on whom legal interception orders have been placed by the authorities.\",\n \"When a terminal (say T1 in FIG.\",\n \"4) wishes to establish an IP connection to another terminal it must first obtain the IP address of the other terminal T2.If the initiating terminal T1 does not know this IP address from the outset, it must obtain it, e.g.\",\n \"by sending a user entered URL to a Domain Name Server (ENS) which can resolve the address and return it to the initiating terminal.\",\n \"A already outlined above, in order to secure the connection, the initiating terminal T1 must then negotiate a pair of IPSec SAs with the other (or “destination”) terminal T2.FIG.\",\n \"5 illustrates the exchange of signalling messages required to establish a pair of SAs.\",\n \"Firstly, the initiating terminal T1 sends a Request ISAKMP SA message to its access network 1.This message contains the identity of the initiating and destination terminals (which may be the terminals' IMSI codes or telephone numbers).\",\n \"The request is handled by the access network's interception server S1.The server checks whether or not it holds the private key for initiating terminal T1.If not, the call connection attempt is rejected.\",\n \"If the server does hold the appropriate private key, the server S1 handles the negotiation of the ISAKMP SA (IKE phase 1) on behalf of the terminal T1 (the terminal T1 does not require IKE Phase 1 capabilities).\",\n \"In the same way, the interception server 52 in the access network 2 of the “destination” terminal T2 handles the ISAKMP SA negotiation on behalf of that terminal T2.This is possible because the interception servers S1,S2 know the private keys of the respective terminals T1,T2.Upon completion of the negotiation, the relevant ISAKMP SA parameters are returned by the interception servers to their respective terminals.\",\n \"As the access networks are private networks, typically using security protocols to secure transmission over the radio interface, there is no danger of the returned parameters falling into the wrong hands.\",\n \"(As an alternative to IKE Phase 1, Kerberos authentication may be used.)\",\n \"At this stage, the interception servers S1,S2 know the parameters which will be used to secure the subsequent negotiation of IPSec SAs between the terminals (i.e.\",\n \"IKE Phase 2).\",\n \"When terminal T1 wishes to begin a secure communication with terminal T2, it sends a Begin IPSec SA negotiation request to terminal T2.This message is intercepted by the access network 1 and the interception server S1 is notified.\",\n \"The request includes in its header portion the relevant ISAKMP SA data.\",\n \"The server S1 then checks its records to see if either the initiating or destination terminals have legal interception orders placed on them.\",\n \"If they do not, the IPSec SA negotiation continues without the involvement of interception server S1.It is noted that once the IPSec SAs have been negotiated in this way, it is not possible for the access network to intercept (and decrypt) traffic carried using the LPSec SAs.\",\n \"If on the other hand the interception server S1 determines that a legal interception order has been placed on one of the terminals T1 or T2, the server S1 interposes itself between terminals T1 and T2 during the negotiation of the pair of IPSec SAs (similarly server S2 may interpose itself in the IPSec SA negotiation if that server decides that it too must intercept the connection).\",\n \"As far as terminals T1 and T2 are concerned, they are negotiating the IPSec SAs with each other and are not aware of the role of the server(s) in the process.\",\n \"Subsequently, encrypted traffic may be intercepted at the server S1, decrypted using the negotiated IPSec SA parameters known to server S1, and routed to an appropriate monitoring point.\",\n \"An important feature is that neither of the terminals is allowed to run the fill ISAKMP negotiation.\",\n \"There is illustrated in FIG.\",\n \"6 signalling associated with an alternative interception scheme.\",\n \"In this scheme, successful legal interception requires the active participation of both interception servers S1,S2 which again hold the private keys of their respective subscribers, and also assumes that the terminals involved in the connection are capable of handling the IKE phase 1 negotiations (nb.\",\n \"this latter requirement does not exist for the embodiment of FIG.\",\n \"5).\",\n \"When an interception server (S1 in FIG.\",\n \"6) receives a request for an ISAKMP SA negotiation from an originating terminal T1, that interception server S1 forwards the request to the destination side interception server S2.If the initiating side interception server S1 determines that legal interception is required, a flag is set in the sent request.\",\n \"When the destination side interception server S2 receives the request it checks the status of the interception flag.\",\n \"If that flag is set, or if the receiving interception server S2 determines from its own records that legal interception is required, the server S2 performs the first ISAKMP SA negotiation (including the Diffie-Hellman exchange) directly with the initiating terminal T1 on behalf of the destination terminal T2.The destination side interception server S2 also notifies the originating side server S1 that it should negotiate a second ISAKMP SA directly with the destination terminal T2.When the initiating side access network 1 subsequently intercepts an IPSec SA negotiation request sent from the initiating terminal T1 (and intended for the destination terminal T2), the request is forwarded to the destination side interception server S2.The destination side interception server S2 represents itself to terminal T1 as if it were the destination terminal T2 for the purpose of the IPSec phase 2 negotiation.\",\n \"A pair of IPSec SAs are established between terminal T1 and the interception server S2.At the same time, the destination side interception server S2 instructs the initiating side interception server S1 to negotiate a second pair of IPSec SA with the destination terminal T2.Server S1 therefore represents itself to the destination terminal T2 as if it were terminal T1 during the IPSec phase 2 negotiation.\",\n \"A second pair of IPSec-SAs are established between server S1 and terminal T2.It is necessary to echo data between the servers S1 and S2 in order to give the servers access to both channels.\",\n \"Thus, data originating from terminal T1 is reflected by server S2 to server S1.Server S1 then encrypts the data using the appropriate IPSec SA and sends it to terminal T2.Similarly, data originating from terminal T2 is reflected by server S1 to server S2, with server S2 encrypting the data using the appropriate IPSec SA and sending it to terminal T1.It is noted that with the embodiment of FIG.\",\n \"6 there is no need for “artificial” ISAKMP SA transfers (between servers and terminals).\",\n \"Rather, the initiating terminals IKE signalling will be processed by the destination side interception server and vice versa.\",\n \"It will be appreciated by the person of skill in the art that various modifications may be made to the above described embodiments without departing from the scope of the present invention.\",\n \"For example, in the embodiment described with reference to FIG.\",\n \"6, the additional delay which results from echoing one monitored channel back from one interception server to the other may be avoided by executing a secure IKE proxy server at the interception server which is monitoring the connection.\",\n \"This proxy server is sent from the other interception server and contains the private key of the terminal for which it is responsible.\",\n \"The private key is incorporated into the proxy server in such a way that it is not disclosed to the receiving interception server.\",\n \"However, when the proxy server is executed, it allows the interception server to conduct the IKE phase 1 and 2 negotiations directly with the initiating and destination terminals.\",\n \"Thus, the interception server will have access to both traffic channels.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473964"}}},{"rowIdx":771,"cells":{"text":{"kind":"list like","value":[["Preparation containing polydimethylsiloxane for nail, cartilage,bone, joint,muscle and tendon disorders","The present invention relates to a topical preparation that contains polydimethylsiloxane for the external treatment of nail, cartilage, bone, joint, muscle and tendon disorders.","The preparation can also contain biotin, zinc, lysine, methionine, laurel extract, carotene, casein, collagen, glucose, a glucosamine and/or arnica extract.","In particular the invention relates to a combination therapy where a glucosamine is administered orally."],["1.Topical preparation that contains an active ingredient and, as carrier, at least 50% (m/m), based on the weight of the preparation, of polydimethylsiloxane for the external treatment of nail, cartilage, bone, joint, muscle and tendon disorders.","2.Preparation according to claim 1, wherein the polydimethylsiloxane has a viscosity of 10,000 to 100,000 cSt.","3.Preparation according to claim 1, wherein the active ingredient is chosen from biotin, zinc, lysine, methionine and mixtures thereof.","4.Preparation according to claim 3, that contains biotin, zinc, lysine and methionine.","5.Preparation according to claim 1, that further contains a glucosamine, derivatives thereof and/or salts thereof, preferably N-acetyl-D-glucosamine or a pharmaceutically acceptable salt thereof, more preferentially N-acetyl-D-glucosamine HCl.","6.Preparation according to claim 1 that further contains one or more of the following: laurel extract, carotene, casein, collagen, glucose and arnica extract.","7.Preparation according to claim 1, that contains biotin, lysine, methionine, zinc, casein, carotene, arnica extract, laurel extract and N-acetyl-D-glucosamine HCl.","8.Preparation according to claim 1, that contains glucose, collagen, carotene, casein, laurel extract and N-acetyl-D-glucosamine HCl.","9.Preparation according to claim 1, that contains arnica extract.","10.Preparation according to claim 1 for the treatment of horses.","11.Preparation according to claim 10 for the treatment of disorders of the hooves, feet and lower legs of horses.","12.Preparation according to claim 11, for the treatment of horses' hooves.","13.Preparation according to claim 1 for the treatment of humans.","14.Use of an oral preparation that contains a glucosamine, derivatives thereof and/or salts thereof in combination with the preparation according to claim 1 for the treatment of nail, cartilage, bone, joint, muscle and tendon disorders.","15.Use according to claim 14, wherein the oral preparation contains N-acetyl-Dglucosamine or a pharmaceutically acceptable salt thereof, preferably N-acetyl-Dglucosamine HCl.","16.Kit comprising an oral preparation that contains a glucosamine and a topical preparation that contains polydimethylsiloxane as a combination for simultaneous, separate or successive administration.","17.Kit according to claim 16, for the treatment of nail, cartilage, bone, joint, muscle and tendon disorders.","18.Use of polydimethylsiloxane having a viscosity of 10,000 to 100,000 cSt as carrier in a topical preparation, wherein the preparation contains at least 50% (m/m), based on the weight of the preparation, of polydimethylsiloxane.","19.Use according to claim 18, wherein the topical preparation is used for the external treatment of nails, cartilage, bones, joints, muscles and tendons.","20.Use according to claim 18, wherein the topical preparation contains at least 50% (m/m) polydimethylsiloxane."],["The present invention relates to a preparation for the treatment of nail, cartilage, bone, joint, muscle and tendon disorders.","In particular, the invention relates to a preparation for the treatment of lower legs, feet and hooves of horses.","The treatment of nail, cartilage, bone, joint, muscle and tendon disorders is generally carried out by combating the disorder internally or externally at the site of the disorder.","In WO 00/15202, for example, it is described that nail disorders can be treated by topical application of an agent that contains an active ingredient in combination with a carrier based on a C1-C4 alkyl ester of lactic acid, malic acid, tartaric acid or citric acid.","Antibiotics and disinfectants can, for example, be incorporated as active ingredients.","U.S. Pat.","No.","4,996,043 describes a composition for the treatment of keratinous material, in particular horses' hooves, which contains an aqueous dispersion containing 20-90% water, oils with a high content of mono-unsaturated fatty acids and water-soluble or -swellable polysaccharides.","Furthermore, a maximum of 10% silicone oil can be added.","The disadvantage of the preparations according to the state of the art is that they do not provide adequate transfer of the active component to the surface to be treated.","As a result the active component does not penetrate adequately.","Furthermore, in particular in the case of the treatment of horses, it is important that the preparation remains on the location to be treated for some considerable time and is not substantially removed by, for example, walking in pasture.","It has now been found that nail, cartilage, bone, joint, muscle and tendon disorders can be effectively prevented and/or treated by the use of a preparation that contains an active ingredient and, as carrier, at least 50% (m/m), based on the weight of the preparation, of polydimethylsiloxane.","Polydimethylsiloxane, which is also termed dimethicone or dimethylsilicone, is a compound having the following structural formula: It is known as an agent for the protection of the skin, for example in the case of decubitus, incontinence and dermatoses.","It has now been found that polydimethylsiloxane is an excellent carrier in the case of the treatment of the abovementioned disorders.","In particular this agent increases the permeability of the tissue to which it is applied, as a result of which the action of active components that are present in the agent is increased.","As illustrated in the examples, experiments with 30 horses suffering from diverse tendon and hoof disorders have shown, according to the protocol, that an appreciable improvement in the condition of the horses was obtained using the preparation according to the claims.","Preferably, a polydimethylsiloxane having a viscosity of 10,000 to 100,000 cSt (mm2/s), preferably 40,000 to 80,000 cSt, is used.","The preparation preferably contains at least 70% (m/m) polydimethylsiloxane.","Furthermore, the preparation is essentially anhydrous, that is to say it contains less than 20% (m/m), preferably less than 10% (m/m), and most preferentially less than 5% (m/m) water.","It is also possible to use an activated polydimethylsiloxane, for example activated with silicon dioxide (simethicone).","The percentage of silicon dioxide can then be approximately 2 to approximately 20% (m/m), based on the total preparation.","The active ingredients incorporated in the preparation of the invention are preferably one or more of the following: biotin, zinc, lysine and methionine, preferably a combination of all these substances.","Biotin (hexahydro-2-oxo-1H-thienol[3,4-d]imidazole-4-pentanoic acid) is a growth factor which is present in small amounts in every cell.","The quantity of biotin in the preparation according to the invention is 10 to 100 gram per 400 gram polydimethylsiloxane.","Biotin is a basic raw material for the production of keratin (the hoof in horses and the nails in man).","Furthermore, stress is reduced, as a result of which reconditioning proceeds more efficiently.","Stress has the effect of destroying the growth process.","Lysine and methionine are essential amino acids.","The preparation according to the invention contains 1 to 20 gram lysine and 1 to 20 gram methionine per 400 gram polydimethylsiloxane.","Together with biotin, lysine has an effect on the formation of keratin.","Together with methionine, lysine forms keratin from the epithelium.","Zinc is present in the preparation in an amount of 1 to 10 gram zinc per 400 gram polydimethylsiloxane.","Zinc acts as a catalyst which in combination with biotin, lysine and methionine forms and produces keratin in the epithelium under the influence of oxygen-rich blood.","In addition to the abovementioned active ingredients, one or more of the following can also be added to the preparation: laurel extract, carotene, caseinate, collagen, glucose and arnica extract.","Laurel extract is obtained from the laurel berry, Laurus nobilis L. The laurel extract can be in the form of a laurel powder, but can also be a laurel oil.","The laurel extract is obtained in the manner known in the art.","The preparation according to the invention contains 1 to 30 gram of laurel extract per 400 gram polydimethylsiloxane.","Laurel extract promotes the growth maturing from keratin.","It also prevents premature aging of tissue, for example in horses in the further outgrowth region of the hoof, it guarantees an improvement in the elasticity of the horn wall.","Carotene, in particular β-carotene, is also referred to as provitamin A.","It occurs in, for example, carrots and palm oil.","In horses, for example, carotene indirectly and directly produces improved keratin production in the growth region of the hoof, at the transition from the leathery skin to the hoof horn.","The preparation according to the invention contains 1 to 20 gram carotene per 400 gram polydimethylsiloxane.","Caseinate or casein is a mixture of phosphorus proteins and has a high content of essential amino acids.","It can be obtained from milk in the conventional manner.","Preferably (calcium) caseinate from cow's milk is used.","The preparation contains 1 to.","20 gram caseinate per 400 gram polydimethylsiloxane.","Collagen is a polypeptide which comprises one third of the total protein in vertebrate mammals.","It is one of the most important constituents of the skin, tissue and the organic component in bones and teeth.","The preparation contains 5 to 30 gram collagen per 400 gram polydimethylsiloxane.","The topical preparation preferably also contains a glucosamine, derivatives thereof and/or salts, which gives particularly favourable results.","Preferably an acetylglucosamine is used and more preferentially N-acetyl-D-glucosamine or a pharmaceutically acceptable salt thereof, in particular N-acetyl-D-glucosamine HCl.","An alternative component that can be incorporated in the topical preparation is arnica extract.","This is obtained from Arnica montana L. Arnica oil is preferably used in the preparation according to the invention.","The quantity of arnica extract is 1 to 10 gram per 400 gram polydimethylsiloxane.","It reduces pain in the case of injuries and improves the recovery process.","According to another aspect of the invention three separate topical preparations are provided.","The first preparation comprises polydimethylsiloxane and one or more, preferably all, of the following: biotin, lysine, methionine, zinc, caseinate, carotene, laurel powder and glucosamine.","This first preparation is in particular suitable for the treatment of nails, hooves, cartilage and joints.","The second preparation comprises polydimethylsiloxane and one or more, preferably all, of the following: glucose, collagen, carotene, caseinate, laurel powder and glucosamine.","This second preparation is in particular suitable for the treatment of tendons and muscles.","The third preparation contains polydimethylsiloxane and arnica extract.","This third preparation is in particular suitable for pain control.","The nail, cartilage, bone, joint, muscle and tendon disorders which can be treated using the preparation according to the invention can exist both in humans and in animals.","Examples of disorders in humans are rheumatic disorders such as Cushing's syndrome, osteoporosis, rheumatoid arthritis and arthrosis, cartilage degeneration, necrosis, joint disorders, joint mobilisation and impaired growth.","The topical agent is applied to the intended location at least once a week.","In addition it has been found that the preparation can advantageously be used in the case of tendon and ligament disorders.","It has been found that the preparation according to the invention is particularly suitable for the treatment of disorders in horses, in particular disorders of the hooves, feet and lower legs, most preferentially of hooves.","Examples of disorders which can be treated using the preparation according to the invention are: loose wall, an interruption in the union between the supporting rim of the horn wall and the horn sole in the white line or in some other location; hollow wall, detachment between various laminae of the wall; horny pillar, a cylindrical renewal of the horn in the longitudinal direction on the inside of the horny wall; horn cracks, to be differentiated according to location, extent and depth.","They mostly occur on the horn wall.","The horn cracks in the wall run in the longitudinal direction, that is to say in the direction of the horn bars.","There are said to be bearing surface cracks, coronet cracks and continuous cracks, from the coronet to the bearing surface; navicular disease, inflammation of the navicular bone, the collective name for frog, mucus bursa and coffin bone flexor tendon.","Chronic inflammation of this area is termed chronic navicular disease (Podotrochleitis chronica); crumbly hoof; hard hooves; red frog; cancer of the frog; and hoof cancer.","In addition to the topical application of the abovementioned preparation based on polydimethylsiloxane, it has been found that combination with an oral preparation that contains a glucosamine, derivatives thereof and/or salts gives particularly favourable results.","Preferably, an acetylglucosamine, more preferentially N-acetyl-D-glucosamine or a pharmaceutically acceptable salt thereof, in particular N-acetyl-D-glucosamine HCl, is used.","The quantity of glucosamine per day is 100 to 2,500 mg, preferably 1,000 to 2,000 mg.","The combination of the topical preparation with a glucosamine is in particular suitable when improvement in the cartilage and protection and build up of the joint is desired.","In addition this combination can be used to improve the function or reconditioning of tendons and muscles.","The oral preparation that contains a glucosamine can be, for example, a capsule, tablet, powder or drink solution.","It contains a carrier suitable for the type of dosage form.","In particular, the oral preparation that contains a glucosamine is a tablet with, as carrier, one or more of the following: gelatine, magnesium stearate, sugar and lactose.","The topical preparation according to the preparation can have, for example, the following compositions: Preparation 1 (in particular suitable for the treatment of nails, cartilage, bones and hooves) polydimethylsiloxane (dimethicone, viscosity 60,000 cSt) 400 g biotin 60 g lysine 7.5 g methionine 7.5 g zinc 2.3 g caseinate 4 g carotene 2 g laurel powder 8 g N-acetyl-D-glucosamine HCl 1.5 g Preparation 2 (in particular suitable for treatment of joints, muscles and tendons) polydimethylsiloxane (dimethicone, viscosity 60,000 cSt) 400 g glucose syrup 40 g collagen 20 g carotene 10 g caseinate 10 g laurel powder 10 g N-acetyl-D-glucosamine HCl 1.5 g Preparation 3 polydimethylsiloxane (dimethicone, viscosity 60,000 cSt) 400 g arnica oil 6.5 g When treating with the topical preparation the latter is applied to the site of the disorder.","Fresh preparation is applied regularly, for example once a week.","The treatment continues as long as is necessary to heal the disorder.","In general this will be a period of 1 to 6 months.","The oral preparation containing glucosamine is preferably administered in combination with the topical preparation.","The oral preparation is administered daily, for example in a dosage of 1 gram per day.","Optionally this daily dose can be split into several doses.","EXPERIMENTAL Various horses were treated in accordance with the protocol for 6 months with the topical and optionally the oral preparation according to the invention.","An appreciable improvement in the condition of the animal that was treated was found to take place.","A number of cases are described in detail below.","Case 1 Hoof Cracks—Bearing Surface Crack Horse with aging hoof cracks caused by poor elasticity and by poor keratin production by the hoof horn.","Loading stress too great for the poor horn quality.","Clinical picture: uncomfortable gait.","Treatment Smear hoof ointment (Preparation 1) into coronet once a week.","1 gram per day N-acetyl-D-glucosamine HCl, administered orally.","As a result of treatment, development of a good horn structure is taking place in the outgrowth region and the horn elasticity is improving.","The structural change from the outgrowth region to the dying-off region proceeds more regularly.","As a result of the above changes the forces can be relaxed in such a way that the crack does not again continue into the transition region.","The aging crack is starting to disappear.","The stress indication line is disappearing.","Clinical picture: horse moves comfortably.","Treatment Smear hoof ointment (Preparation 1) into coronet once per week.","Oral administration of powder stopped.","As a result of further treatment, good horn structure with more than adequate elasticity.","The dying-off region has become small.","The stress and aging crack is restricted to the dying-off region.","Clinical picture: horse moves comfortably and tactmatically.","Case 2 Hoof Cracks—Coronet Crack The horse has developed hoof cracks as a result of overstressing in combination with poor elasticity of the keratin union of the hoof horn and poor quality of the cartilage in the hoof, resulting in a hoof mechanism that works poorly.","The loading stress is too great, as a result of which the hoof splits open from the coronet.","Clinical picture: walks gingerly.","Start of Treatment Smear hoof ointment (Preparation 1) into coronet once a week.","1 gram per day N-acetyl-D-glucosamine HCl, administered orally.","As a result of treatment the coronet is becoming more supple and the horn elasticity has increased, crack is no longer open and is gradually closing by growth from the inside.","Clinical picture: horse walks comfortably and tactmatically.","Treatment Smear hoof ointment (Preparation 1) into the coronet once per week.","As a result of further treatment the coronet has become supple.","Horn elasticity is good, crack has closed up.","Very good recovery of the coronet.","Clinical picture: horse walks comfortably and tactmatically.","In a corresponding manner, the use of the hoof ointment (Preparation 1), on its own or in combination with the oral preparation, was found to be suitable for the treatment of loose wall, tilting of the coffin bone, horny pillar, seizure of the hoof mechanism and ball arthrosis.","Case 3 Tearing of the Flexor Tendon The flexor tendon of the right hind leg of the horse was ruptured.","Immediately after the tendon was ruptured it was established by means of a tendon scan that only 10% of the tendon was still attached, with a large amount of scar tissue and blood clotting.","Clinical picture: the leg was very swollen and no load was put on it.","Treatment: Smearing with tendon ointment (Preparation 2) and 1 gram per day N-acetyl-D-glucosamine HCl, administered orally.","As a result of the treatment, during the first month the horse cautiously started to put weight on the leg and its ball gave way to a considerable extent (almost down to the ground).","The swelling gradually decreased.","During the second and third months the horse started to put increasing weight on the leg.","The ball also gradually gave way to a lesser extent and the swelling in the leg disappeared.","The horse started to put weight on the leg when at rest.","A further tendon scan was made after the third month.","The tendon was 75% recovered without visible scar or connective tissue."]],"string":"[\n [\n \"Preparation containing polydimethylsiloxane for nail, cartilage,bone, joint,muscle and tendon disorders\",\n \"The present invention relates to a topical preparation that contains polydimethylsiloxane for the external treatment of nail, cartilage, bone, joint, muscle and tendon disorders.\",\n \"The preparation can also contain biotin, zinc, lysine, methionine, laurel extract, carotene, casein, collagen, glucose, a glucosamine and/or arnica extract.\",\n \"In particular the invention relates to a combination therapy where a glucosamine is administered orally.\"\n ],\n [\n \"1.Topical preparation that contains an active ingredient and, as carrier, at least 50% (m/m), based on the weight of the preparation, of polydimethylsiloxane for the external treatment of nail, cartilage, bone, joint, muscle and tendon disorders.\",\n \"2.Preparation according to claim 1, wherein the polydimethylsiloxane has a viscosity of 10,000 to 100,000 cSt.\",\n \"3.Preparation according to claim 1, wherein the active ingredient is chosen from biotin, zinc, lysine, methionine and mixtures thereof.\",\n \"4.Preparation according to claim 3, that contains biotin, zinc, lysine and methionine.\",\n \"5.Preparation according to claim 1, that further contains a glucosamine, derivatives thereof and/or salts thereof, preferably N-acetyl-D-glucosamine or a pharmaceutically acceptable salt thereof, more preferentially N-acetyl-D-glucosamine HCl.\",\n \"6.Preparation according to claim 1 that further contains one or more of the following: laurel extract, carotene, casein, collagen, glucose and arnica extract.\",\n \"7.Preparation according to claim 1, that contains biotin, lysine, methionine, zinc, casein, carotene, arnica extract, laurel extract and N-acetyl-D-glucosamine HCl.\",\n \"8.Preparation according to claim 1, that contains glucose, collagen, carotene, casein, laurel extract and N-acetyl-D-glucosamine HCl.\",\n \"9.Preparation according to claim 1, that contains arnica extract.\",\n \"10.Preparation according to claim 1 for the treatment of horses.\",\n \"11.Preparation according to claim 10 for the treatment of disorders of the hooves, feet and lower legs of horses.\",\n \"12.Preparation according to claim 11, for the treatment of horses' hooves.\",\n \"13.Preparation according to claim 1 for the treatment of humans.\",\n \"14.Use of an oral preparation that contains a glucosamine, derivatives thereof and/or salts thereof in combination with the preparation according to claim 1 for the treatment of nail, cartilage, bone, joint, muscle and tendon disorders.\",\n \"15.Use according to claim 14, wherein the oral preparation contains N-acetyl-Dglucosamine or a pharmaceutically acceptable salt thereof, preferably N-acetyl-Dglucosamine HCl.\",\n \"16.Kit comprising an oral preparation that contains a glucosamine and a topical preparation that contains polydimethylsiloxane as a combination for simultaneous, separate or successive administration.\",\n \"17.Kit according to claim 16, for the treatment of nail, cartilage, bone, joint, muscle and tendon disorders.\",\n \"18.Use of polydimethylsiloxane having a viscosity of 10,000 to 100,000 cSt as carrier in a topical preparation, wherein the preparation contains at least 50% (m/m), based on the weight of the preparation, of polydimethylsiloxane.\",\n \"19.Use according to claim 18, wherein the topical preparation is used for the external treatment of nails, cartilage, bones, joints, muscles and tendons.\",\n \"20.Use according to claim 18, wherein the topical preparation contains at least 50% (m/m) polydimethylsiloxane.\"\n ],\n [\n \"The present invention relates to a preparation for the treatment of nail, cartilage, bone, joint, muscle and tendon disorders.\",\n \"In particular, the invention relates to a preparation for the treatment of lower legs, feet and hooves of horses.\",\n \"The treatment of nail, cartilage, bone, joint, muscle and tendon disorders is generally carried out by combating the disorder internally or externally at the site of the disorder.\",\n \"In WO 00/15202, for example, it is described that nail disorders can be treated by topical application of an agent that contains an active ingredient in combination with a carrier based on a C1-C4 alkyl ester of lactic acid, malic acid, tartaric acid or citric acid.\",\n \"Antibiotics and disinfectants can, for example, be incorporated as active ingredients.\",\n \"U.S. Pat.\",\n \"No.\",\n \"4,996,043 describes a composition for the treatment of keratinous material, in particular horses' hooves, which contains an aqueous dispersion containing 20-90% water, oils with a high content of mono-unsaturated fatty acids and water-soluble or -swellable polysaccharides.\",\n \"Furthermore, a maximum of 10% silicone oil can be added.\",\n \"The disadvantage of the preparations according to the state of the art is that they do not provide adequate transfer of the active component to the surface to be treated.\",\n \"As a result the active component does not penetrate adequately.\",\n \"Furthermore, in particular in the case of the treatment of horses, it is important that the preparation remains on the location to be treated for some considerable time and is not substantially removed by, for example, walking in pasture.\",\n \"It has now been found that nail, cartilage, bone, joint, muscle and tendon disorders can be effectively prevented and/or treated by the use of a preparation that contains an active ingredient and, as carrier, at least 50% (m/m), based on the weight of the preparation, of polydimethylsiloxane.\",\n \"Polydimethylsiloxane, which is also termed dimethicone or dimethylsilicone, is a compound having the following structural formula: It is known as an agent for the protection of the skin, for example in the case of decubitus, incontinence and dermatoses.\",\n \"It has now been found that polydimethylsiloxane is an excellent carrier in the case of the treatment of the abovementioned disorders.\",\n \"In particular this agent increases the permeability of the tissue to which it is applied, as a result of which the action of active components that are present in the agent is increased.\",\n \"As illustrated in the examples, experiments with 30 horses suffering from diverse tendon and hoof disorders have shown, according to the protocol, that an appreciable improvement in the condition of the horses was obtained using the preparation according to the claims.\",\n \"Preferably, a polydimethylsiloxane having a viscosity of 10,000 to 100,000 cSt (mm2/s), preferably 40,000 to 80,000 cSt, is used.\",\n \"The preparation preferably contains at least 70% (m/m) polydimethylsiloxane.\",\n \"Furthermore, the preparation is essentially anhydrous, that is to say it contains less than 20% (m/m), preferably less than 10% (m/m), and most preferentially less than 5% (m/m) water.\",\n \"It is also possible to use an activated polydimethylsiloxane, for example activated with silicon dioxide (simethicone).\",\n \"The percentage of silicon dioxide can then be approximately 2 to approximately 20% (m/m), based on the total preparation.\",\n \"The active ingredients incorporated in the preparation of the invention are preferably one or more of the following: biotin, zinc, lysine and methionine, preferably a combination of all these substances.\",\n \"Biotin (hexahydro-2-oxo-1H-thienol[3,4-d]imidazole-4-pentanoic acid) is a growth factor which is present in small amounts in every cell.\",\n \"The quantity of biotin in the preparation according to the invention is 10 to 100 gram per 400 gram polydimethylsiloxane.\",\n \"Biotin is a basic raw material for the production of keratin (the hoof in horses and the nails in man).\",\n \"Furthermore, stress is reduced, as a result of which reconditioning proceeds more efficiently.\",\n \"Stress has the effect of destroying the growth process.\",\n \"Lysine and methionine are essential amino acids.\",\n \"The preparation according to the invention contains 1 to 20 gram lysine and 1 to 20 gram methionine per 400 gram polydimethylsiloxane.\",\n \"Together with biotin, lysine has an effect on the formation of keratin.\",\n \"Together with methionine, lysine forms keratin from the epithelium.\",\n \"Zinc is present in the preparation in an amount of 1 to 10 gram zinc per 400 gram polydimethylsiloxane.\",\n \"Zinc acts as a catalyst which in combination with biotin, lysine and methionine forms and produces keratin in the epithelium under the influence of oxygen-rich blood.\",\n \"In addition to the abovementioned active ingredients, one or more of the following can also be added to the preparation: laurel extract, carotene, caseinate, collagen, glucose and arnica extract.\",\n \"Laurel extract is obtained from the laurel berry, Laurus nobilis L. The laurel extract can be in the form of a laurel powder, but can also be a laurel oil.\",\n \"The laurel extract is obtained in the manner known in the art.\",\n \"The preparation according to the invention contains 1 to 30 gram of laurel extract per 400 gram polydimethylsiloxane.\",\n \"Laurel extract promotes the growth maturing from keratin.\",\n \"It also prevents premature aging of tissue, for example in horses in the further outgrowth region of the hoof, it guarantees an improvement in the elasticity of the horn wall.\",\n \"Carotene, in particular β-carotene, is also referred to as provitamin A.\",\n \"It occurs in, for example, carrots and palm oil.\",\n \"In horses, for example, carotene indirectly and directly produces improved keratin production in the growth region of the hoof, at the transition from the leathery skin to the hoof horn.\",\n \"The preparation according to the invention contains 1 to 20 gram carotene per 400 gram polydimethylsiloxane.\",\n \"Caseinate or casein is a mixture of phosphorus proteins and has a high content of essential amino acids.\",\n \"It can be obtained from milk in the conventional manner.\",\n \"Preferably (calcium) caseinate from cow's milk is used.\",\n \"The preparation contains 1 to.\",\n \"20 gram caseinate per 400 gram polydimethylsiloxane.\",\n \"Collagen is a polypeptide which comprises one third of the total protein in vertebrate mammals.\",\n \"It is one of the most important constituents of the skin, tissue and the organic component in bones and teeth.\",\n \"The preparation contains 5 to 30 gram collagen per 400 gram polydimethylsiloxane.\",\n \"The topical preparation preferably also contains a glucosamine, derivatives thereof and/or salts, which gives particularly favourable results.\",\n \"Preferably an acetylglucosamine is used and more preferentially N-acetyl-D-glucosamine or a pharmaceutically acceptable salt thereof, in particular N-acetyl-D-glucosamine HCl.\",\n \"An alternative component that can be incorporated in the topical preparation is arnica extract.\",\n \"This is obtained from Arnica montana L. Arnica oil is preferably used in the preparation according to the invention.\",\n \"The quantity of arnica extract is 1 to 10 gram per 400 gram polydimethylsiloxane.\",\n \"It reduces pain in the case of injuries and improves the recovery process.\",\n \"According to another aspect of the invention three separate topical preparations are provided.\",\n \"The first preparation comprises polydimethylsiloxane and one or more, preferably all, of the following: biotin, lysine, methionine, zinc, caseinate, carotene, laurel powder and glucosamine.\",\n \"This first preparation is in particular suitable for the treatment of nails, hooves, cartilage and joints.\",\n \"The second preparation comprises polydimethylsiloxane and one or more, preferably all, of the following: glucose, collagen, carotene, caseinate, laurel powder and glucosamine.\",\n \"This second preparation is in particular suitable for the treatment of tendons and muscles.\",\n \"The third preparation contains polydimethylsiloxane and arnica extract.\",\n \"This third preparation is in particular suitable for pain control.\",\n \"The nail, cartilage, bone, joint, muscle and tendon disorders which can be treated using the preparation according to the invention can exist both in humans and in animals.\",\n \"Examples of disorders in humans are rheumatic disorders such as Cushing's syndrome, osteoporosis, rheumatoid arthritis and arthrosis, cartilage degeneration, necrosis, joint disorders, joint mobilisation and impaired growth.\",\n \"The topical agent is applied to the intended location at least once a week.\",\n \"In addition it has been found that the preparation can advantageously be used in the case of tendon and ligament disorders.\",\n \"It has been found that the preparation according to the invention is particularly suitable for the treatment of disorders in horses, in particular disorders of the hooves, feet and lower legs, most preferentially of hooves.\",\n \"Examples of disorders which can be treated using the preparation according to the invention are: loose wall, an interruption in the union between the supporting rim of the horn wall and the horn sole in the white line or in some other location; hollow wall, detachment between various laminae of the wall; horny pillar, a cylindrical renewal of the horn in the longitudinal direction on the inside of the horny wall; horn cracks, to be differentiated according to location, extent and depth.\",\n \"They mostly occur on the horn wall.\",\n \"The horn cracks in the wall run in the longitudinal direction, that is to say in the direction of the horn bars.\",\n \"There are said to be bearing surface cracks, coronet cracks and continuous cracks, from the coronet to the bearing surface; navicular disease, inflammation of the navicular bone, the collective name for frog, mucus bursa and coffin bone flexor tendon.\",\n \"Chronic inflammation of this area is termed chronic navicular disease (Podotrochleitis chronica); crumbly hoof; hard hooves; red frog; cancer of the frog; and hoof cancer.\",\n \"In addition to the topical application of the abovementioned preparation based on polydimethylsiloxane, it has been found that combination with an oral preparation that contains a glucosamine, derivatives thereof and/or salts gives particularly favourable results.\",\n \"Preferably, an acetylglucosamine, more preferentially N-acetyl-D-glucosamine or a pharmaceutically acceptable salt thereof, in particular N-acetyl-D-glucosamine HCl, is used.\",\n \"The quantity of glucosamine per day is 100 to 2,500 mg, preferably 1,000 to 2,000 mg.\",\n \"The combination of the topical preparation with a glucosamine is in particular suitable when improvement in the cartilage and protection and build up of the joint is desired.\",\n \"In addition this combination can be used to improve the function or reconditioning of tendons and muscles.\",\n \"The oral preparation that contains a glucosamine can be, for example, a capsule, tablet, powder or drink solution.\",\n \"It contains a carrier suitable for the type of dosage form.\",\n \"In particular, the oral preparation that contains a glucosamine is a tablet with, as carrier, one or more of the following: gelatine, magnesium stearate, sugar and lactose.\",\n \"The topical preparation according to the preparation can have, for example, the following compositions: Preparation 1 (in particular suitable for the treatment of nails, cartilage, bones and hooves) polydimethylsiloxane (dimethicone, viscosity 60,000 cSt) 400 g biotin 60 g lysine 7.5 g methionine 7.5 g zinc 2.3 g caseinate 4 g carotene 2 g laurel powder 8 g N-acetyl-D-glucosamine HCl 1.5 g Preparation 2 (in particular suitable for treatment of joints, muscles and tendons) polydimethylsiloxane (dimethicone, viscosity 60,000 cSt) 400 g glucose syrup 40 g collagen 20 g carotene 10 g caseinate 10 g laurel powder 10 g N-acetyl-D-glucosamine HCl 1.5 g Preparation 3 polydimethylsiloxane (dimethicone, viscosity 60,000 cSt) 400 g arnica oil 6.5 g When treating with the topical preparation the latter is applied to the site of the disorder.\",\n \"Fresh preparation is applied regularly, for example once a week.\",\n \"The treatment continues as long as is necessary to heal the disorder.\",\n \"In general this will be a period of 1 to 6 months.\",\n \"The oral preparation containing glucosamine is preferably administered in combination with the topical preparation.\",\n \"The oral preparation is administered daily, for example in a dosage of 1 gram per day.\",\n \"Optionally this daily dose can be split into several doses.\",\n \"EXPERIMENTAL Various horses were treated in accordance with the protocol for 6 months with the topical and optionally the oral preparation according to the invention.\",\n \"An appreciable improvement in the condition of the animal that was treated was found to take place.\",\n \"A number of cases are described in detail below.\",\n \"Case 1 Hoof Cracks—Bearing Surface Crack Horse with aging hoof cracks caused by poor elasticity and by poor keratin production by the hoof horn.\",\n \"Loading stress too great for the poor horn quality.\",\n \"Clinical picture: uncomfortable gait.\",\n \"Treatment Smear hoof ointment (Preparation 1) into coronet once a week.\",\n \"1 gram per day N-acetyl-D-glucosamine HCl, administered orally.\",\n \"As a result of treatment, development of a good horn structure is taking place in the outgrowth region and the horn elasticity is improving.\",\n \"The structural change from the outgrowth region to the dying-off region proceeds more regularly.\",\n \"As a result of the above changes the forces can be relaxed in such a way that the crack does not again continue into the transition region.\",\n \"The aging crack is starting to disappear.\",\n \"The stress indication line is disappearing.\",\n \"Clinical picture: horse moves comfortably.\",\n \"Treatment Smear hoof ointment (Preparation 1) into coronet once per week.\",\n \"Oral administration of powder stopped.\",\n \"As a result of further treatment, good horn structure with more than adequate elasticity.\",\n \"The dying-off region has become small.\",\n \"The stress and aging crack is restricted to the dying-off region.\",\n \"Clinical picture: horse moves comfortably and tactmatically.\",\n \"Case 2 Hoof Cracks—Coronet Crack The horse has developed hoof cracks as a result of overstressing in combination with poor elasticity of the keratin union of the hoof horn and poor quality of the cartilage in the hoof, resulting in a hoof mechanism that works poorly.\",\n \"The loading stress is too great, as a result of which the hoof splits open from the coronet.\",\n \"Clinical picture: walks gingerly.\",\n \"Start of Treatment Smear hoof ointment (Preparation 1) into coronet once a week.\",\n \"1 gram per day N-acetyl-D-glucosamine HCl, administered orally.\",\n \"As a result of treatment the coronet is becoming more supple and the horn elasticity has increased, crack is no longer open and is gradually closing by growth from the inside.\",\n \"Clinical picture: horse walks comfortably and tactmatically.\",\n \"Treatment Smear hoof ointment (Preparation 1) into the coronet once per week.\",\n \"As a result of further treatment the coronet has become supple.\",\n \"Horn elasticity is good, crack has closed up.\",\n \"Very good recovery of the coronet.\",\n \"Clinical picture: horse walks comfortably and tactmatically.\",\n \"In a corresponding manner, the use of the hoof ointment (Preparation 1), on its own or in combination with the oral preparation, was found to be suitable for the treatment of loose wall, tilting of the coffin bone, horny pillar, seizure of the hoof mechanism and ball arthrosis.\",\n \"Case 3 Tearing of the Flexor Tendon The flexor tendon of the right hind leg of the horse was ruptured.\",\n \"Immediately after the tendon was ruptured it was established by means of a tendon scan that only 10% of the tendon was still attached, with a large amount of scar tissue and blood clotting.\",\n \"Clinical picture: the leg was very swollen and no load was put on it.\",\n \"Treatment: Smearing with tendon ointment (Preparation 2) and 1 gram per day N-acetyl-D-glucosamine HCl, administered orally.\",\n \"As a result of the treatment, during the first month the horse cautiously started to put weight on the leg and its ball gave way to a considerable extent (almost down to the ground).\",\n \"The swelling gradually decreased.\",\n \"During the second and third months the horse started to put increasing weight on the leg.\",\n \"The ball also gradually gave way to a lesser extent and the swelling in the leg disappeared.\",\n \"The horse started to put weight on the leg when at rest.\",\n \"A further tendon scan was made after the third month.\",\n \"The tendon was 75% recovered without visible scar or connective tissue.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473968"}}},{"rowIdx":772,"cells":{"text":{"kind":"list like","value":[["Method for transferring information and associated network transition units","Disclosed is a method wherein operative, administrative and maintenance functions are defined initially for a time-division multiplex oriented communication network (12).","In order to utilize said functions in a packet-oriented communication network (10), they are emulated by said network (10)."],["1.A method for transferring information within at least one time-division multiplex oriented communication network via at least one packet-oriented communication network, comprising: implementing functions for operation and/or administration and/or maintenance in the time-division multiplex oriented communication network for transfer of the information within the time-division multiplex oriented communication network; transferring at least a part of the information via the packet-oriented communication network; and emulating at least some of the functions for operation, administration and maintenance of the time-division multiplex oriented communication network by the packet-oriented communication network.","2.The method according to claim 1, wherein a customer-side network transition unit is disposed between a customer area and the packet-oriented communication network, and an exchange-side network transition unit is disposed between the packet-oriented communication network and an exchange of the time-division multiplex oriented communication network.","3.The method according to claim 2, wherein a function relates to the activation of a loopback from the exchange via the exchange-side network transition unit back to the exchange, the exchange sets at least one Sa5 bit to the value zero and using successive Sa6 bits transmits an Sa6 bit sequence with the value “1111” at least eight times, the values of the Sa5 bits and the Sa6 bit sequence are detected in the exchange-side network transition unit, when the value zero is detected for the Sa5 bit when the value “1111” is detected eight times in succession for the Sa6 bit sequence by the exchange-side network transition unit, the data arriving from the exchange is transmitted back essentially unchanged to the exchange, whereby at least one A bit with the value one and at least one Sa5 bit with the value zero are transmitted from the exchange-side network transition unit to the exchange, and at least one A bit with the value one is transferred by the exchange-side network transition unit via the packet-oriented communication network.","4.The method according to claim 2, wherein one of the functions relates to activation of a loopback from the exchange, via the exchange-side network transition unit, via the customer-side network transition unit, back to the exchange-side network transition unit and then back to the exchange, the exchange transmits at least one Sa5 bit with a value zero and an Sa6 bit sequence with a value “1010” at least eight times, the value of the Sa5 bit and the value of the Sa6 bit sequence are detected in the exchange-side network transition unit, when the value zero is detected for the Sa5 bit and when the value “1010” is detected eight times in succession for the Sa6 bit sequence by the exchange-side network transition unit, the data arriving from the exchange is transmitted back essentially unchanged to the exchange, whereby at least one A bit with a value one and at least one Sa5 bit with a value zero are transmitted by the exchange-side network transition unit to the exchange, and whereby the incoming data is manipulated depending on transmission errors in a part of the loopback located in the packet-oriented communication network, the transmission of loopback data packets is initiated by the exchange-side network transition unit, the loopback data packets are detected in the customer-side network transition unit and transmitted back to the exchange-side network transition unit, the exchange-side network transition unit checks the received loopback data packets and depending on the result of the check manipulates the data to be transmitted to the exchange, and upon detection of a loopback data packet at least one A bit with the value one is transmitted to a customer telephony equipment by the customer-side network transition unit.","5.The method according to claim 2, wherein one of the functions relates to the monitoring of the performance and continuity of the transmission from the exchange-side network transition unit to the customer-side network transition unit, performance data packets and continuity data packets are generated in the exchange-side network transition unit and transmitted to the customer-side network transition unit, the performance data packets and continuity data packets are monitored in the customer-side network transition unit, when an error is detected by the customer-side network transition unit at least one E bit with a value zero is transmitted via the packet-oriented communication network to the exchange-side network transition unit and is forwarded to the exchange, and in the exchange-side network transition unit a new checksum is computed for the data to be transmitted to the exchange.","6.The method according to claim 2, wherein on of the function relates to the monitoring of the performance and continuity of the transmission from the customer-side network transition unit to the exchange-side network transition unit, performance data packets and continuity data packets are generated in the customer-side network transition unit and transmitted to the exchange-side network transition unit, the performance data packets and continuity data packets are monitored in the exchange-side network transition unit, upon an error the computation of a checksum for the data to be transmitted to the exchange is manipulated, and in the error-free case a checksum for the data to be transmitted to the exchange is computed without manipulation.","7.The method according to claim 2, wherein one of the function relates to an error which is reported by a customer telephony equipment, that the error monitoring method is performed in the customer telephony equipment, when an error is detected by the customer telephony equipment least one E bit with a value zero is transmitted to the customer-side network transition unit, the value of the E bit is monitored in the customer-side network transition unit, when the value zero is detected for the E bit by the customer-side network transition unit at least one Sa5 bit with the value one and at least one Sa6 bit sequence with the value “0001” are transmitted via the packet-oriented communication network to the exchange-side network transition unit and are forwarded to the exchange.","8.The method according to claim 2, wherein one of the function relates to an error which is detected by the customer-side network transition unit, the error monitoring method is performed in the customer-side network transition unit, when an error is detected by the customer-side network transition unit at least one Sa5 bit with a value one and at least one Sa6 bit sequence with a value “0010” are transmitted via the packet-oriented communication network to the exchange-side network transition unit and are forwarded to the exchange, and upon an error being detected by the customer-side network transition unit at least one E bit with the value zero is transmitted to the customer telephony equipment.","9.The method according to claim 2, wherein one of the function relates to a first error which is reported by a customer telephony equipment and to a second error which is detected by the customer-side network transition unit, a first error monitoring method is performed in the customer telephony equipment, upon an error being detected by the customer telephony equipment at least one E bit with a value zero is transmitted to the customer-side network transition unit, the values of the E bits are monitored in the customer-side network transition unit, a second error monitoring method is performed in the customer-side network transition unit, and upon an error being detected by the second error monitoring method and when the value zero is detected for the E bit by the customer-side network transition unit at least one Sa5 bit with a value one and at least one Sa6 bit sequence with a value “0011” are transmitted via the packet-oriented communication network to the exchange-side network transition unit and are forwarded to the exchanges.","10.The method according to acclaim 2, wherein one of the function relates to the detection of a loss of signal or a loss of frame alignment between a customer telephony equipment and the customer-side network transition unit, upon a loss being detected in the customer-side network transition unit at least one Sa5 bit with a value one and at least one Sa6 bit sequence with a value “1100” are transmitted by the customer-side network transition unit via the packet-oriented communication network to the exchange-side network transition unit and are forwarded to the exchange, and when a loss is detected by the customer-side network transition unit at least one E bit with the value zero is transmitted to the customer telephony equipment.","11.The method according to claim 1, wherein one of the functions relates to detection of a loss of continuity or a loss of a cell or a loss of cell alignment or a physical connection failure in the packet-oriented communication network, continuity data packets are generated in the exchange-side network transition unit and transmitted to the customer-side network transition unit, the continuity data packets are detected in the customer-side network transition unit, upon non-arrival of the continuity data packets or upon a loss of cell or a loss of cell alignment or a physical connection failure being detected by the customer-side network transition unit, a continuous bit sequence of bits with a value one is transmitted to a customer telephony equipment, and upon non-arrival of the continuity data packets or upon a loss of cell or a loss of cell alignment or a physical connection failure being detected in the customer-side network transition unit A bits originating from the customer telephony equipment are transmitted unchanged via the packet-oriented communication network, and upon non-arrival of the continuity data packets or upon a loss of cell or a loss of cell alignment or a physical connection failure being detected by the customer-side network transition unit at least one Sa5 bit with a value one and at least one Sa6 bit sequence with a value “1110” are transmitted via the packet-oriented communication network to the exchange-side network transition unit and are forwarded to the exchange.","12.The method according to acclaim 2, wherein one of the functions relates to detection of a loss of signal between the exchange and the exchange-side network transition unit, upon a loss being detected in the exchange-side network transition unit by the exchange-side network transition unit no further continuity data packets are transmitted via the packet-oriented communication network, the continuity data packets are monitored in the customer-side network transition unit, upon non-arrival of the continuity data packets a continuous bit sequence of bits with a value one is transmitted by the customer-side network transition unit to a customer telephony equipment, upon non-arrival of the continuity data packets in the customer-side network transition unit A bits originating from the customer telephony equipment are transmitted unchanged via the packet-oriented communication network, and upon non-arrival of the continuity data packets from the customer-side network transition unit at least one Sa5 bit with a value one and at least one Sa6 bit sequence with a value “1110” are transmitted via the packet-oriented communication network to the exchange-side network transition unit and are forwarded to the exchange.","13.The method according to claim 1, wherein one of the functions relates to detection of a loss of signal or a loss of frame alignment between a customer telephony equipment and the customer-side network transition unit and to detection of a loss of signal between the exchange and the exchange-side network transition unit, upon a loss of signal being detected between the exchange and the exchange-side network transition unit in the exchange-side network transition unit no further continuity data packets are transmitted via the packet-oriented communication network, the continuity data packets are monitored in the customer-side network transition unit, upon non-arrival of the continuity data packets a continuous bit sequence of bits with a value one is transmitted by the customer-side network transition unit to the customer telephony equipment, upon non-arrival of the continuity data packets at least one Sa5 bit with a value one and at least one Sa6 bit sequence with a value “1110” are transmitted by the customer-side network transition unit via the packet-oriented communication network to the exchange-side network transition unit and from there are forwarded to the exchange, and upon a loss of signal or a loss of frame alignment being detected between the customer telephony equipment and the customer-side network transition unit and simultaneous non-arrival of the continuity data packets at least one A bit with a value zero is transmitted by the customer-side network transition unit via the packet-oriented communication network to the exchange-side network transition unit and is forwarded to the exchange.","14.The method according to claim 2, wherein one of the function relates to detection of a loss of continuity or a loss of cell or a loss of cell alignment or a physical connection failure between the customer-side network transition unit and the exchange-side network transition unit, continuity data packets are generated in the customer-side network transition unit and transmitted to the exchange-side network transition unit, upon non-arrival of the continuity data packets or upon a loss of cell or a loss of cell alignment or a physical connection failure being detected by the exchange-side network transition unit a bit sequence conforming to a predefined auxiliary bit pattern is transmitted to the exchange, and the exchange, after receiving the bit sequence, transmits an A bit with the value one to the exchange-side network transition unit.","15.The method according to acclaim 2, wherein one of the function relates to detection of a continuous bit sequence of bits with a value one, an occurrence of the continuous bit sequence of bits with a value one originating from the exchange is monitored in the exchange-side network transition unit, that when the bit sequence is detected at least one alarm data packet is generated and transmitted via the packet-oriented communication network to the customer-side network transition unit, the occurrence of alarm data packets is detected in the customer-side network transition unit, when an alarm data packet is detected by the customer-side network transition unit the continuous bit sequence of bits with a value one is transmitted to a customer telephony equipment, when an alarm data packet is detected in the customer-side network transition unit A bits originating from the customer telephony equipment are transmitted unchanged via the packet-oriented communication network, and when an alarm data packet is detected by the customer-side network transition unit at least one Sa6 bit sequence with a value “1111” is transmitted via the packet-oriented communication network to the exchange-side network transition unit and is forwarded to the exchange.","16.The method according to claim 2, wherein on of the functions relates to detection of a continuous bit stream of bits with a value one and to detection of a loss of signal or a loss of frame alignment between a customer telephony equipment and the customer-side network transition unit, an occurrence of the continuous bit sequence of bits with a value one originating from the exchange is monitored in the exchange-side network transition unit, when the bit sequence is detected at least one alarm data packet is generated and transmitted via the packet-oriented communication network to the customer-side network transition unit, the occurrence of alarm data packets and the signal and the frame alignment between the customer telephony equipments and the customer-side network transition unit are monitored in the customer-side network transition unit, when an alarm data packet tis detected by the customer-side network transition unit the continuous bit sequence of bits with a value one is transmitted to the customer telephony equipment, when an alarm data packet and a loss of signal or a loss of frame alignment is detected, at least one A bit with a value zero is transmitted by the customer-side network transition unit via the packet-oriented data communication network to the exchange-side network transition unit and from there is forwarded to the exchanges, and when an alarm data packet is detected by the customer-side network transition unit at least one Sa5 bit with a value one and at least one Sa6 bit sequence with a value “1111” are transmitted via the packet-oriented communication network to the exchange-side network transition unit and are forwarded to the exchange.","17.The method according to claim 2, wherein one of the functions relates to detection of a voltage failure in the customer-side network transition unit, the operating voltage is monitored in the customer-side network transition unit, upon failure of the operating voltage in the customer-side network transition unit at least one Sa6 bit sequence with a value “1000” is transmitted via the packet-oriented communication network to the exchange-side network transition unit and is forwarded to the exchange, upon failure of the operating voltage in the customer-side network transition unit at least one E bit with a value zero is transmitted to a customer telephony equipment, and the exchange-side network transition unit transmits the Sa6 bit sequence unchanged and subsequently a bit sequence conforming to a predefined auxiliary bit pattern to the exchange.","18.The method according to claim 2, wherein the emulation is implemented by functions of the packet-oriented communication network.","19.The method according to claim 2, wherein the packet-oriented communication network is implemented according to asynchronous transfer mode.","20.The method according to claim 2, wherein the packet-oriented communication network is implemented according to the asynchronous transfer mode Adaption Layer 1 or according to the asynchronous transfer mode Adaption Layer 2.21.The method according to claim 2, wherein the packet-oriented communication network is implemented according to the Internet Protocol.","22.The method according to claim 2, wherein the time-division multiplex oriented communication network is an ISDN network.","23.The method according to claim 22, wherein part of a primary multiplex access is replaced by the packet-oriented communication network, and/or the primary multiplex access includes an XDSL line between the customer-side network transition unit and the packet-oriented communication network.","24.The method according to claim 2, wherein operation, administration and maintenance functions are provided essentially according to ETSI standard ETS 300 011 and/or according to ITU-T standard G.962 and/or according to ETSI standard ETS 300 233.25.The method according to claim 2, wherein a multi time frame including sixteen time frames according to ITU-T standard G.704 is used in the time-division multiplex oriented communication network, the multi time frame including an A bit in a third position and/or an Sa5 bit in a fifth position and/or an Sa6 bit in a sixth position in a start time slot of every second time frame beginning with the second start time slot of the multi time frame, and/or including an E bit in the first position of the second-to-last time frame and/or the last time frame of a multi time frame.","26.The method according to claim 25, wherein the data of the start time slot of each time frame is transmitted unchanged in value via the packet-oriented communication network.","27.The method according claim 2, wherein located between a customer telephony equipment and the time-division multiplex oriented communication network a broadband subscriber line via which primary multiplex traffic is carried.","28.","(Canceled) 29.","(Canceled)"],["<SOH> BACKGROUND OF THE INVENTION <EOH>The time-division multiplex oriented communication network is, for example, an ISDN data transmission network (ISDN—Integrated Services Digital Network).","In the time-division multiplex oriented communication network the data is transferred in different time slots in accordance with a time-division multiplex method.","The packet-oriented communication network is a network in which the information or data is transferred in data packets.","The packet-oriented communication network is for example a communication network operating according to the Internet Protocol.","Another example of a packet-oriented communication network is an ATM network (ATM—Asynchronous Transfer Mode), in which, however, the data packets are referred to as cells.","Functions for operation, administration and/or maintenance in the time-division multiplex oriented communication network have been defined for example in the following standards of the ETSI (European Telecommunications Standards Institute) or ITU-T (International Telecommunication Union—Telecommunication Standardization Sector): ETSI ETS 300 233, Integrated Services Digital Network (ISDN); Access Digital Section for ISDN Primary Rate, May 1994, ITU-T G.962, Digital Sections and Digital Line Systems; Access Digital Section for ISDN Primary Rate at 2048 kbit/s, March 1993, ETSI ETS 300 011, Integrated Services Digital Network (ISDN); Primary Rate User-Network Interface Layer 1 Specification and Test Principles, April 1992.Thus, the functions for operation, administration and/or maintenance relate for example to the activation of loopbacks or to error monitoring."],["<SOH> SUMMARY OF THE INVENTION <EOH>The invention relates to a method for transferring information within at least one time-division multiplex oriented communication network via at least one packet-oriented communication network.","Functions for operation and/or administration and/or maintenance are implemented in the time-division multiplex oriented communication network with the aim of transferring information within the time-division multiplex oriented communication network.","The information relates for example to user data or voice data.","The invention discloses, in one embodiment, a method for transmitting information within at least one time-division multiplex oriented communication network via at least one packet-oriented communication network, the method continuing to allow functions for operation, administration and/or maintenance of the time-division multiplex oriented communication network to be used in the time-division multiplex oriented communication network.","Associated network transition units are also to be specified.","In another embodiment according to the invention, at least some of the information is transferred via the packet-oriented communication network.","At the same time at least some of the functions for operation, administration and/or maintenance are emulated by the packet-oriented communication network.","As a result the functions specified for the time-division multiplex oriented communication network can continue to be used essentially without restriction.","In another embodiment according to the invention, a customer-side network transition unit is interposed between a customer area and the packet-oriented communication network.","The customer-side network transition unit is also referred to as a CP-IWF (Customer Premises—Interworking Function).","In the embodiment, an exchange-side network transition unit is additionally interposed between the packet-oriented communication network and an exchange of the time-division multiplex oriented communication network.","The exchange-side network transition unit is also referred to as a CO-IWF (Central Office—Interworking Function).","In other embodiments according to the invention individual functions for operation, administration and maintenance are.","In this case, the functions known from the time-division multiplex oriented communication network are retained in the time-division multiplex oriented communication network and emulated in the packet-oriented communication network.","In still another embodiment according to the invention, the emulation is provided by functions of the packet-oriented communication network, in particular by the customer-side network transition unit and by the exchange-side network transition unit.","In another embodiment, the packet-oriented communication network is an ATM network which is implemented according to the asynchronous transfer mode (ATM).","By means of this embodiment powerful methods which have been defined in ATM standards, for example in the standards of the ATM Forum, can be used: af-vtoa-0113.000, ATM Trunking Using AAL-2 for Narrowband Services, February 1999, af-vmoa-0145.000, Voice and Multimedia over ATM—Loop Emulation Service Using AAL-2, July 2000.In another embodiment, the packet-oriented communication network is implemented according to the ATM Adaption Layer 1 or according to the ATM Adaption Layer 2.These layers are also referred to as AAL1 and AAL2 respectively (AAL—ATM Adaption Layer).","The AAL2 layer in particular is well suited to a reduction in bandwidth during the transmission in the packet-oriented communication network, in particular for the connection of subscribers.","Reference is made for example to the standard af-vmoa-0145.000, Section 2.3, CP-IWF ATM Interfaces, 2.3.1 Physical Layer, where XDSL methods (XDSL—X-Digital Subscriber Line) are cited as examples of transmission methods, i.e.","the ADSL method (ADSL—Asymmetrical Digital Subscriber Line) and the SDSL method (SDSL—Symmetrical Digital Subscriber Line).","In yet another embodiment the packet-oriented communication network is implemented according to the Internet Protocol.","In this way, for example, the IP over ATM transmission method can be used.","In another embodiment, the time-division multiplex oriented communication network is an ISDN network.","In connection with the invention the following standards relating to the ISDN network are particularly relevant: ITU-T I.411, ISDN User Network Interfaces—Reference Configuration, ITU-T I.412, ISDN User Network Interfaces—Interface Structures and Access Capabilities, and ITU-T Q.512, Exchange Interfaces for Subscriber Access.","In another embodiment, parts of a primary multiplex access are replaced by the packet-oriented communication network.","The primary multiplex access is also referred to as a Primary Rate Access.","In one embodiment, the primary multiplex access is a so-called E1 access with a transmission capacity of 2,048 Mbit/s or a so-called DS1 access with a transmission capacity of 1,544 Mbit/s.","The three above-mentioned standards relate to an E1 access.","Integrating the packet-oriented communication network with the primary multiplex access allows the transmission bandwidth required on a subscriber access line to be considerably reduced.","This enables transmission over two-wire copper lines, for example according to an XDSL method.","In one embodiment, furthermore, a multi time frame consisting of sixteen time frames, as described for example in ITU-T standard G.704, is used in the time-division multiplex oriented communication network.","This time frame is explained in more detail below with reference to FIG.","2 .","The time frame includes in particular a so-called A bit for indicating alarm conditions, a so-called Sa5 bit, a so-called Sa6 bit and a so-called E bit for indicating error conditions.","The bits are transferred in the start time slots TSO of each time frame.","In another embodiment, the start time slot TSO of each time frame is transferred over the packet-oriented communication network unchanged except for the mentioned changes in value, i.e.","in particular without speech compression and without silence suppression.","Another embodiment relates to a customer-side network transition unit and to an exchange-side network transition unit, each of which contain function units during the operation of which the method steps related to the customer-side network transition unit and to the exchange-side network transition unit are performed according to the invention or one of its embodiments.","Thus, the above-mentioned technical effects also apply to the two network transition units."],["CLAIM FOR PRIORITY This application claims priority to International Application No.","PCT/EP02/03753, which was published in the German language on Oct. 17, 2002, which claims the benefit of priority to German Application No.","01108481.1 which was filed in the German language on Apr.","4, 2001.TECHNICAL FIELD OF THE INVENTION The invention relates to a method for transferring information and associated network transition units, and in particular, a method for transferring information within at least one time-division multiplex oriented communication network via at least one packet-oriented communication network.","BACKGROUND OF THE INVENTION The time-division multiplex oriented communication network is, for example, an ISDN data transmission network (ISDN—Integrated Services Digital Network).","In the time-division multiplex oriented communication network the data is transferred in different time slots in accordance with a time-division multiplex method.","The packet-oriented communication network is a network in which the information or data is transferred in data packets.","The packet-oriented communication network is for example a communication network operating according to the Internet Protocol.","Another example of a packet-oriented communication network is an ATM network (ATM—Asynchronous Transfer Mode), in which, however, the data packets are referred to as cells.","Functions for operation, administration and/or maintenance in the time-division multiplex oriented communication network have been defined for example in the following standards of the ETSI (European Telecommunications Standards Institute) or ITU-T (International Telecommunication Union—Telecommunication Standardization Sector): ETSI ETS 300 233, Integrated Services Digital Network (ISDN); Access Digital Section for ISDN Primary Rate, May 1994, ITU-T G.962, Digital Sections and Digital Line Systems; Access Digital Section for ISDN Primary Rate at 2048 kbit/s, March 1993, ETSI ETS 300 011, Integrated Services Digital Network (ISDN); Primary Rate User-Network Interface Layer 1 Specification and Test Principles, April 1992.Thus, the functions for operation, administration and/or maintenance relate for example to the activation of loopbacks or to error monitoring.","SUMMARY OF THE INVENTION The invention relates to a method for transferring information within at least one time-division multiplex oriented communication network via at least one packet-oriented communication network.","Functions for operation and/or administration and/or maintenance are implemented in the time-division multiplex oriented communication network with the aim of transferring information within the time-division multiplex oriented communication network.","The information relates for example to user data or voice data.","The invention discloses, in one embodiment, a method for transmitting information within at least one time-division multiplex oriented communication network via at least one packet-oriented communication network, the method continuing to allow functions for operation, administration and/or maintenance of the time-division multiplex oriented communication network to be used in the time-division multiplex oriented communication network.","Associated network transition units are also to be specified.","In another embodiment according to the invention, at least some of the information is transferred via the packet-oriented communication network.","At the same time at least some of the functions for operation, administration and/or maintenance are emulated by the packet-oriented communication network.","As a result the functions specified for the time-division multiplex oriented communication network can continue to be used essentially without restriction.","In another embodiment according to the invention, a customer-side network transition unit is interposed between a customer area and the packet-oriented communication network.","The customer-side network transition unit is also referred to as a CP-IWF (Customer Premises—Interworking Function).","In the embodiment, an exchange-side network transition unit is additionally interposed between the packet-oriented communication network and an exchange of the time-division multiplex oriented communication network.","The exchange-side network transition unit is also referred to as a CO-IWF (Central Office—Interworking Function).","In other embodiments according to the invention individual functions for operation, administration and maintenance are.","In this case, the functions known from the time-division multiplex oriented communication network are retained in the time-division multiplex oriented communication network and emulated in the packet-oriented communication network.","In still another embodiment according to the invention, the emulation is provided by functions of the packet-oriented communication network, in particular by the customer-side network transition unit and by the exchange-side network transition unit.","In another embodiment, the packet-oriented communication network is an ATM network which is implemented according to the asynchronous transfer mode (ATM).","By means of this embodiment powerful methods which have been defined in ATM standards, for example in the standards of the ATM Forum, can be used: af-vtoa-0113.000, ATM Trunking Using AAL-2 for Narrowband Services, February 1999, af-vmoa-0145.000, Voice and Multimedia over ATM—Loop Emulation Service Using AAL-2, July 2000.In another embodiment, the packet-oriented communication network is implemented according to the ATM Adaption Layer 1 or according to the ATM Adaption Layer 2.These layers are also referred to as AAL1 and AAL2 respectively (AAL—ATM Adaption Layer).","The AAL2 layer in particular is well suited to a reduction in bandwidth during the transmission in the packet-oriented communication network, in particular for the connection of subscribers.","Reference is made for example to the standard af-vmoa-0145.000, Section 2.3, CP-IWF ATM Interfaces, 2.3.1 Physical Layer, where XDSL methods (XDSL—X-Digital Subscriber Line) are cited as examples of transmission methods, i.e.","the ADSL method (ADSL—Asymmetrical Digital Subscriber Line) and the SDSL method (SDSL—Symmetrical Digital Subscriber Line).","In yet another embodiment the packet-oriented communication network is implemented according to the Internet Protocol.","In this way, for example, the IP over ATM transmission method can be used.","In another embodiment, the time-division multiplex oriented communication network is an ISDN network.","In connection with the invention the following standards relating to the ISDN network are particularly relevant: ITU-T I.411, ISDN User Network Interfaces—Reference Configuration, ITU-T I.412, ISDN User Network Interfaces—Interface Structures and Access Capabilities, and ITU-T Q.512, Exchange Interfaces for Subscriber Access.","In another embodiment, parts of a primary multiplex access are replaced by the packet-oriented communication network.","The primary multiplex access is also referred to as a Primary Rate Access.","In one embodiment, the primary multiplex access is a so-called E1 access with a transmission capacity of 2,048 Mbit/s or a so-called DS1 access with a transmission capacity of 1,544 Mbit/s.","The three above-mentioned standards relate to an E1 access.","Integrating the packet-oriented communication network with the primary multiplex access allows the transmission bandwidth required on a subscriber access line to be considerably reduced.","This enables transmission over two-wire copper lines, for example according to an XDSL method.","In one embodiment, furthermore, a multi time frame consisting of sixteen time frames, as described for example in ITU-T standard G.704, is used in the time-division multiplex oriented communication network.","This time frame is explained in more detail below with reference to FIG.","2.The time frame includes in particular a so-called A bit for indicating alarm conditions, a so-called Sa5 bit, a so-called Sa6 bit and a so-called E bit for indicating error conditions.","The bits are transferred in the start time slots TSO of each time frame.","In another embodiment, the start time slot TSO of each time frame is transferred over the packet-oriented communication network unchanged except for the mentioned changes in value, i.e.","in particular without speech compression and without silence suppression.","Another embodiment relates to a customer-side network transition unit and to an exchange-side network transition unit, each of which contain function units during the operation of which the method steps related to the customer-side network transition unit and to the exchange-side network transition unit are performed according to the invention or one of its embodiments.","Thus, the above-mentioned technical effects also apply to the two network transition units.","BRIEF DESCRIPTION OF THE DRAWINGS Exemplary embodiments of the invention are explained in the following with reference to the attached drawings, in which: FIG.","1 shows an ISDN primary multiplex interface model with processing of a time slot TSO according to ETSI standard ETS 300 233.FIG.","2 shows a CRC-4 multi time frame structure.","FIG.","3 shows function units and function sequences for activating a loopback in an exchange-side network transition unit CO-IWF.","FIG.","4 shows function units and function sequences for activating a loopback in a customer-side network transition unit CP-IWF.","FIG.","5 shows the monitoring of an ATM network by means of VCC (Virtual Channel Connection) performance monitoring (PMo—Performance Monitoring) and continuity monitoring (CC—Continuity Check) at an ATMdown interface of the customer-side network transition unit CP-IWF.","FIG.","6 shows the monitoring of the ATM network by means of VCC performance monitoring and continuity monitoring at an ATMup interface of the exchange-side network transition unit CO-IWF.","FIG.","7 shows function sequences when a CRC-4 error (Cyclic Redundancy Check) is reported by a customer telephony equipment TE.","FIG.","8 shows function sequences when a CRC-4 error is detected in a Tup signal of the customer-side network transition unit CP-IWF.","FIG.","9 shows function sequences when a CRC-4 error is reported by the customer equipment TE and simultaneously a CRC-4 error is detected in the Tup signal of the customer-side network transition unit CP-IWF.","FIG.","10 shows function sequences when a loss of signal (LOS—Loss of Signal) is detected or when a loss of frame alignment or frame synchronization (LFA—Loss of Frame Alignment) is detected at the Tup interface of the customer-side network transition unit CP-IWF.","FIG.","11 shows function sequences when an ATM connection error is detected at the ATMdown interface of the customer-side network transition unit CP-IWF.","FIG.","12 shows function sequences when an ATM connection error occurs at the ATMdown interface as a result of a loss of signal (LOS) at the exchange-side network transition unit CO-IWF.","FIG.","13 shows function sequences when a loss of signal (LOS) or loss of frame alignment (LFA) occurs at the T reference point of the customer-side network transition unit CP-IWF and simultaneously a loss of signal (LOS) occurs at the V3 reference point of the exchange-side network transition unit CO-IWF.","FIG.","14 shows function sequences when an ATM connection error occurs in the ATMup signal at the exchange-side network transition unit CO-IWF.","FIG.","15 shows function sequences when an AIS (Alarm Indication Signal) is detected at the customer-side network transition unit CP-IWF.","FIG.","16 shows function sequences when an AIS is detected in the ATMdown signal and simultaneously a loss of signal (LOS) or loss of frame alignment (LFA) occurs at the T reference point of the customer-side network transition unit CP-IWF.","FIG.","17 shows function sequences when an operating voltage failure is detected in the customer-side network transition unit CP-IWF.","FIG.","18 shows the reference model for the subscriber line (loop) emulation service using AAL2.FIG.","19 shows a protocol reference model for a customer-side network transition unit CP-IWF with a user-side ISDN PRI and with DSS1 forwarding (DSS1—Digital Signaling System Number One) via the AAL2 between the network transition units IWF.","FIG.","20 shows a protocol reference model for an exchange-side network transition unit CO-IWF with user-side ISDN PRI and with DSS1 forwarding as well as with L3 (Layer 3) monitoring via the AAL2 between the network transition units IWF.","DETAILED DESCRIPTION OF THE INVENTION Introduction to the Technical Environment There are many reasons why support for an ISDN primary multiplex interface via the AAL2 (ATM Adaption Layer) using the principles and methods of the subscriber line emulation services (LES—Loop Emulation Services) according to the af-vmoa-0145.000 standard is desirable.","A concept for supporting an ISDN primary multiplex interface via AAL2/LES (ATM Adaption Layer 2/Loop Emulation Service) is disclosed in the following.","Also, a text for supplementing the subscriber line emulation service (LES—Loop Emulation Service) using the AAL2 is proposed according to an exemplary embodiment based on the af-vxnoa-0145.000 standard, wherein the ISDN primary multiplex interface (PRI—Primary Rate Interface) is defined as a user-side interface.","At the same time consistency with the support for the ISDN basic access interface (BRI—Basic Rate Interface) via AAL2 is ensured, as explained in the af-vmoa-0145.000 standard.","The aim is to specify a solution that advantageously uses the powerful methods which have been defined for the subscriber line (loop) emulation service and which to date are not to be found in the approach for channel group transmission via AAL2 (AAL2 trunking).","These powerful mechanisms include support for an embedded operation channel (EOC—Embedded Operation Channel), flexible assignment of a channel identifier (CID—AAL2 Channel Identifier) as well as channel activation using the ELCP protocol (ELCP—Emulated Loop Control Protocol).","ISDN PRI Overview An overview of the use of the ISDN primary multiplex interface in the world of classical time-division multiplex (TDM—Time Division Multiplex) is given below.","Also specified are requirements which must be met if parts of the access digital section (DS—Digital Section) of the primary multiplex access are replaced by the ATM network.","Furthermore, methods which meet these requirements are specified.","There are differences between the so-called DS1 (Digital Signal Level Number One) and E1 structures (E1—European Digital Signal Level Number One).","The exemplary embodiments and explanations relate to the E1 structure.","General statements applicable to both interfaces are however made wherever possible.","ISDN Primary Multiplex Interface Model (PRI) with Operating and Maintenance Functions (for a 2048 kbps signal/E1) FIG.","1 shows an access digital section DS with its delimitations and the processing of time slot TS0.The operating and maintenance functions support methods and information elements which are required for control of the access digital section by an exchange ET or a service node.","An Sa5, Sa6, E and A bit of a time slot TS0 are relevant for indication and control purposes.","The bit structure of the time slot TS0 and the multi time frame structure are defined according to ITU-T standard G.704 and are explained in more detail below with reference to FIG.","2.The A bit is used for passing alarm status information between the service node and a customer telephony equipment TE (customer telephony end equipment).","The A bit requires monitoring and is transferred transparently.","Other control bits of the time slot TSO are to be transferred transparently.","Also shown in FIG.","1 are the elements of the access digital section DS which use CRC methods 4, 6 (CRC—Cyclic Redundancy Check).","The CRC-4 methods 4, 6 are implemented and used between the exchange ET and the network termination unit NT1 as well as between the network termination unit NT1 and the customer telephony equipment TE.","This is also known as Option Two according to ITU-T standard G.962.FIG.","1 also shows a line termination unit LT.","Between the line termination unit LT and the exchange ET there is a V3 reference point.","Between the customer telephony equipment TE and the network termination unit NT1 there is a T reference point.","The following table shows the signals which are exchanged between the T reference point and the access digital section DS during the normal operating conditions and error conditions specified in ETSI standard ETS 300 011: Name List of signals Normal operating mode Operating time frame with: time frame active assigned CRC bits, CRC error information (cf.","ITU-T standard G.704 for 2048 Kbit/s systems), no error indications RAI (Remote Alarm Indication) Operating time frame with: active assigned CRC bits, CRC error information (cf.","ITU-T standard G.704 for 2048 Kbit/s systems), remote alarm indication (cf.","ITU-T standard G.704, Table 4a, for 2048 Kbit/s systems, A bit is set to the value one) LOS (Loss of Signal) No input signal received (LOS) AIS (Alarm Indication Signal) Continuous bit stream of bits with the value one CRC error information E bit according to ITU-T standard G.704, Table 4b, set to the value zero if an errored CRC block is received (for a 2048 Kbit/s system only) The signals exchanged between the access digital section DS and the exchange ET are specified in the following table: Name List of signals Normal operating mode Operating time frame with: time frame active assigned CRC bits, CRC error information (cf.","ITU-T standard G.704 for 2048 Kbit/s systems), no error indicators RAI (Remote Alarm Indication) Operating time frame with: active assigned CRC bits, CRC error information (cf.","ITU-T standard G.704 for 2048 Kbit/s systems), remote alarm indication (cf.","ITU-T standard G.704, Table 4a, for 2048 Kbit/s systems, A bit is set to the value one) LOS (Loss of Signal) No input signal received (LOS) AIS (Alarm Indication Signal) Continuous bit stream of bits with the value one CRC error information E bit according to ITU-T standard G.704, Table 4b, set to the value zero if an errored CRC block is received (for a 2048 Kbit/s system only) The following additional signals are required in order to indicate error conditions which occur in relation to the access digital section DS: Name List of signals Normal time frames These are time frames without error indications or loopback requests generated by the exchange ET or the customer telephony equipment TE, where an A bit with the value one or zero is not relevant to the access digital section DS.","Time frames These are time frames which, in addition to the normal time frames, have Sa6 bits containing error indication signals which have been generated in the network termination unit NT1 and transmitted to the exchange ET. Alternatively, the Sa6 bits may contain loopback requests which have been transmitted from the exchange ET to the access digital section DS.","In this case the Sa5 bit is also used to indicate the direction and as a loopback indication.","The Sa5 bit in the transmission direction from the access digital section DS to the exchange ET is set in the network termination unit NT1 or in the line termination unit LT and transmitted to the exchange ET according to the following rules: Sa5 = 1 Loopback 2 not activated, Sa5 = 0 Loopback 2 activated.","The Sa6 bits are numbered Sa6(1), Sa6(2), Sa6(3), Sa6(4) and synchronized with the sub- multi time frames which are explained in more detail below with reference to FIG.","3.Substitute time frame In the event of a loss of signal (LOS) or loss of frame alignment (LFA) at the T reference point of the network termination unit NT1, a new time frame must be generated.","The A bit is set to the value zero and the Sa4, Sa5, Sa7 and Sa8 bits as well as the bits of the time slots TS1 to TS31 are set to the value one.","A bit sequence of Sa6 bits is used to indicate this error condition.","LFA Loss of Frame Alignment Operating voltage failure in the network termination unit NT1 or the line termination unit LT Auxiliary bit pattern This is a time frame less and (AUXP—Auxiliary Pattern) continuous bit sequence of bits with alternating values of one and zero ( .",".",".","101010 .",".",". )","which is transmitted in both transmission directions by the line termination unit LT if a loss of signal (LOS) is detected at the corresponding receive unit FIG.","2 shows the structure of a CRC-4 multi time frame.","The CRC-4 multi time frame consists of two sub-multi time frames I and II.","Each sub-multi time frame I, II consists of eight time frames.","The CRC-4 checksum is computed for all the bits of a sub-multi time frame I, II.","The CRC bits C1, C2, C3 and C4 are transferred in bit one of the frame alignment signal.","The frame alignment signal is transmitted in even-numbered time frames (0, 2, 4, .",".",".",").","The Sa bits, like the A bits (Remote Alarm Indication), are part of the signal not relating to the frame alignment.","The Sa bits and the A bit are transmitted in the odd-numbered time frames (1, 3, 5, .",".",".",").","The first bit in each of the odd-numbered time frames 1, 3, 5, 7, 9 and 11 forms the CRC multi time frame alignment signal, which takes the form of a bit sequence with the value “001011”.","Bit one in the thirteenth time frame is the so-called E bit and serves to indicate the reception of an errored sub-multi time frame by setting the E bit from the value one to the value zero for each errored sub-multi time frame I.","The first bit in the fifteenth time frame serves to indicate an error for each errored sub-multi time frame II.","CRC-4 Method The CRC method serves to provide protection against errored frames (framing) and for error performance monitoring.","This includes the multi time frame methods which are specified in ITU-T standard G.704.If the access digital section DS is replaced by the ATM network 10 and the advantages of the AAL2 are used, it is not possible to perform or implement a CRC-4 method between the exchange ET and the network termination unit NT1.For this reason ATM methods are used for this purpose in the exemplary embodiments, although these methods do not offer the same possibilities as CRC monitoring.","VCC (Virtual Channel Connection) performance monitoring (PMo—Performance Monitoring) and continuity monitoring (continuity check) are to be used in both transmission directions for monitoring between the network transition unit NT1 and the line termination unit LT. A new computation according to the CRC-4 error monitoring method is performed in the line termination unit LT in the transmission direction toward the exchange ET. No changes are necessary for the CRC-4 method between the network termination unit NT1 and the customer telephony equipment TE.","Operation and Maintenance of the Access Digital Section The methods described in the following permit the emulation of an operation and maintenance method for a primary multiplex interface (PRI OAM—Primary Rate Interface Operation Administration Maintenance) via an interface with the subscriber line (loop) emulation service (LES Interface—Loop Emulation Service Interface).","The access digital section DS provides the means for transmitting indicator elements and for detecting error conditions at the T reference point interface and the V3 reference point interface as well as for supporting test methods.","The following functions are supported according to ETSI standard ETS 300 233: Loopbacks Loopback 1, transparent loopback in the line termination unit LT or in the exchange-side network transition unit CO-IWF (F1), Loopback 2, transparent loopback in the network termination unit NT1 or in the customer-side network transition unit CP-IWF (F2), Error conditions within the access digital section DS loss of signal (LOS) or loss of frame alignment (LFA) on the line side of the network termination unit NT1 or the customer-side network transition function CP-IWF (in the the downstream signal; that is, the signal coming from the line termination unit LT or exchange-side network transition unit CO-IWF to the network termination unit NT1 or the customer-side network transition unit CP-IWF) (F9), loss of signal (LOS) on the line side of the line termination unit LT or the exchange-side network transition unit CO-IWF (F12), operating voltage failure in the network termination unit NT1 or in the customer-side network transition unit CP-IWF (F13), AIS (Alarm Indication Signal) on the line side of the network termination unit NT1 or the customer-side network transition unit CP-IWF, the AIS being generated in the network and forwarded transparently by the line termination unit LT or the exchange-side network transition unit CO-IWF (F13), at the V3 reference point loss of signal (LOS} (F10), at the T reference point loss of signal (LOS) or loss of frame alignment (LFA) (F8), operating voltage failure (if relevant) (F-) Error performance monitoring errored CRC blocks detected on the line side of the network termination unit NT1 or the customer-side network transition unit CP-IWF (F3), errored CRC blocks detected at the T reference point of the network termination unit NT1 or the customer-side network transition unit CP-IWF (F6), CRC error indication received from the customer telephony equipment TE in the E bit (F5), errored CRC blocks detected at the T reference point of the network termination unit NT1 or the customer-side network transition unit CP-IWF and simultaneous reception of a CRC error information from the customer telephony equipment TE (F7).","The functions referred to are explained in more detail below with reference to exemplary embodiments.","With regard to the above-mentioned functions, the reference specified in parentheses in each case indicates which emulated function F1 to F15 is used for the function concerned.","The emulated functions F1 to F15 are explained in more detail below in this order with reference to FIGS.","3 to 17.The functions F4, F11 and F14 are not referred to in the above-mentioned list.","The error condition on which the function F11 is based is an overlay composed of the error conditions on which functions F8 and F10 are based.","The error condition on which function F14 is based is an overlay comprised of the error conditions on which functions F8 and F13 are based.","During a normal operating mode of the access digital section and in error conditions which permit the use of time frame signals, i.e.","excluding loss of signal (LOS) or loss of frame alignment (LFA) at the T reference point of the network termination unit NT1 or the customer-side network transition unit CP-IWF, the information is transferred transparently in the time slots of the ISDN PRI signal via the access digital section D5.Similarly, the A, Sa4, Sa7 and Sa8 bits are also transferred transparently.","The time frame and multi time frame alignment, the CRC-4 bits and the CRC error information (E bit) in both transmission directions as well as the Sa5 and Sa6 bits in the transmission direction toward the exchange ET are generated in the network termination unit NT1 or the customer-side network transition unit CP-IWF.","In the case of a loss of signal (LOS) or a loss of time frame alignment (LFA) at the T reference point of the network termination unit NT1 or the customer-side network transition unit CP-IWF, a new time frame is generated.","The A bit is set to the value zero.","The Sa4, Sa5, Sa7 and Sa8 bits as well as the bits in the time slots TS1 to TS31 are set to the value one.","These time frames are also known as substituted frames.","A bit sequence consisting of Sa6 bits is used to indicate this error condition.","In the exemplary embodiment, instead of a full time-division multiplex frame (TDM Frame) being transferred via the ATM section, each time slot is mapped into a separate AAL2 channel.","The time slots TS1 to TS31 contain user information and DSS1 signaling (DSS1—Digital Signaling System Number One).","The time slot TS0 transfers error conditions or control information, e.g.","loopback requests.","As mentioned already, some of the classical PRI-OAM methods (PRI-OAM—Primary Rate Interface-Operation Administration Maintenance) have to be emulated by means of ATM methods, whereby standardized methods defined in ITU-T standard I.610 are used.","Thus, for example, the CRC-4 methods, the event indication and/or error indication in the access digital section DS and also the loopback methods are emulated by means of suitable ATM methods.","The generation of the time frame and the generation of the multi time frame alignment signal are not possible in the network termination unit NT1 or the customer-side network transition unit CP-IWF in the transmission direction toward the exchange ET.","The line termination unit LT or the exchange-side network transition unit CO-IWF handles these functions in the transmission direction toward the exchange ET.","In the transmission direction toward the customer telephony equipment TE, the network termination unit NT1 or the customer-side network transition unit CP-IWF generates the time frame, the multi time frame alignment, the CRC-4 bits and the CRC error indication or error information.","General Explanations Relating to FIGS.","3 to 18 In the upper part of each of the FIGS.","3 to 17 there is shown a table in the top row of which the columns of a table header are assigned in each case to the following elements in the order given below: column 1 to the customer telephony equipment TE, which is also referred to as a CPE (Customer Premises Equipment), column 2 to the T reference point, columns 3 and 4 to the customer-side network transition unit CP-IWF, column 5 to the ATM network 10, columns 6 and 7 to the exchange-side network transition unit CO-IWF, column 8 to the reference point V3, and column 10 to the exchange ET, which is also referred to as a service node (Switching Node).","Rows 2 and 3 of the table header relate to the assignment of the columns in the transmission direction from the customer telephony equipment TE to the exchange ET. For this transmission direction: column 1 relates to the data transmitted by the customer telephony equipment 12, column 3 relates to the data received in the customer-side network transition unit CP-IWF from the side of the customer telephony equipment TE, column 4 relates to the data transmitted by the customer-side network transition unit CP-IWF into the ATM network 10, column 6 relates to the data received in the exchange-side network transition unit CO-IWF from the ATM network 10, column 7 relates to the data transmitted by the exchange-side network transition unit CO-IWF to the exchange ET, and column 9 relates to the data received in the exchange ET from the exchange-side network transition unit CO-IWF.","The positions mentioned are also referred to in this order as the Tup, Tup, ATMup, ATMup, V3up and V3up interface.","Rows 2 and 3 of the table header are in each case assigned to the first row of the table body.","Rows 4 and 5 of the table header relate to the transfer of data in the transmission direction from the exchange ET to the customer telephony equipment TE.","For this transmission direction: the first column of the table relates to the data received in the customer telephony equipment TE and originating from the customer-side network transition unit CP-IWF, column 3 relates to the data transmitted by the customer-side network transition unit CP-IWF to the customer telephony equipment TE, column 4 relates to the data received by the customer-side network transition unit CP-IWF from the ATM network 10, column 6 relates to the data transmitted by the exchange-side network transition unit CO-IWF into the ATM network 10, column 7 relates to the data received in the exchange-side network transition unit CO-IWF from the exchange ET, and column 9 relates to the data transmitted by the exchange ET to the exchange-side network transition unit CO-IWF.","The positions mentioned for this transmission direction are also referred to in the given order as the Tdown, Tdown, ATMdown, ATMdown, V3down and V3down interface.","The second or last row of the table body is assigned to the transmission direction from the exchange ET to the customer telephony equipment TE.","The following symbols are used in the fields of the tables: - no entry because none is necessary to an understanding of the exemplary embodiment or because in accordance with the known standard, a analyze, t transmit unchanged underscore error/event detection point x= .",".",".","response to an error/event condition ### error/event cause In addition to the entities already explained with reference to FIG.","1, FIGS.","3 to 17 also show a time-division multiplex data transmission network 12 on both sides of the ATM network 10.Loopbacks Two different loopbacks are supported, a loopback in the exchange-side network transition unit CO-IWF and another loopback in the customer-side network transition unit CP-IWF.","The loopbacks are activated as a result of special Sa bit sequences which are explained in more detail below with reference to FIGS.","3 and 4.A bit sequence with eight successive code words of the bits Sa6(1), Sa6(2), Sa6(3) and Sa6(4) forms a loopback command which must be detected before further measures are taken.","Conversely, the loopback is released if eight successive commands for releasing the loopback or eight other successive signals have been received in which no loopback command has been detected.","FIG.","3 shows function units and function sequences for activating a loopback in the exchange-side network transition unit CO-IWF.","In the case of the loopback in the exchange-side network transition unit CO-IWF, the exchange-side network transition unit CO-IWF transmits the ISDN PRI signal back to the exchange ET.","In the transmission direction toward the customer-side network transition unit CP-IWF, the A bit is set to the value one.","In the transmission direction toward the exchange ET, the A bit is set to the value one and the Sa5 bit is set to the value zero.","The function explained with reference to FIG.","3 is also referred to as function F1.FIG.","4 shows function units and function sequences for activating a loopback in the customer-side network transition unit CP-IWF.","The loopback in the customer-side network transition unit CP-IWF is produced as a result of the insertion of end-to-end F5 loopback cells which are also referred to as F5 LB (Loopback) cells.","These cells are inserted by the exchange-side network transition unit CO-IWF after the loopback command relating to the network termination unit NT or the customer-side network transition unit CP-IWF has been detected.","At the same time there is set up in the exchange-side network transition unit CO-IWF a local loopback which is similar to the loopback in the case explained with reference to FIG.","3 of a loopback in the line termination unit LT or the exchange-side network transition unit CO-IWF.","The loopback is regarded as faulty if the LB cells do not return to their point of origin within five seconds.","In this case the exchange-side network transition unit CO-IWF manipulates the loopback signal transmitted to the exchange ET. As a result the exchange ET can detect bit errors.","The function explained with reference to FIG.","4 is also referred to as function F2.Error Performance Monitoring The CRC-4 monitoring is used to provide protection against incorrect time frames (framing) and for error performance monitoring of the access digital section DS.","In order to enable service features such as free channel suppression, speech compression, voice activity detection and silence suppression, transparent transmission of the CRC-4 is not possible.","For this reason ATM monitoring methods such as VCC (Virtual Channel Connection) performance monitoring and VCC continuity monitoring are used simultaneously for the ATM section of the access digital section DS.","The performance monitoring ensures real-time evaluation of the transmission quality for the selected VCCs at segment level or at end-to-end level.","In the case of monitoring of the ATM section of the access digital section DS, the end-to-end VCC performance monitoring variant should be used.","The performance monitoring is achieved by monitoring blocks from user cells.","There are two applications of performance monitoring, namely forward monitoring and backward reporting together or forward monitoring on its own.","In order to detect errored blocks of user cells it is sufficient to use only forward monitoring.","Performance monitoring is activated during the connection setup as part of the VCC method.","In addition the activation or deactivation can be configured for each VCC with the aid of a telecommunication management network (TMN).","For communication between the VCC end points, special OAM cells (OAM—Operation Administration Maintenance) are used, namely the FPM cells (FPM—Forward Performance Monitoring) and the BR cells (BR—Backward Reporting).","The following parameters are obtained with the aid of performance monitoring: a counter value for the number of transferred user information cells, errored blocks, a counter value for lost user information cells and a counter value for erroneously inserted user information cells within a monitored block consisting of cells.","The CRC-4 checksum is computed afresh at the point of transition from the ATM network 10 to the time-division multiplex oriented network 12, i.e.","in the transmission direction toward the exchange ET.","If errors are detected as a result of VCC performance monitoring and continuity monitoring in the exchange-side network transition unit CO-IWF, this results in a CRC-4 error at the V3up interface of the exchange-side network transition unit CO-IWF (see explanatory remarks relating to FIG.","6 below).","If a block error is detected by the exchange-side network transition unit CO-IWF, the E bit in the transmission direction toward the exchange ET is set to the value zero.","This case is explained in FIG.","5.FIG.","5 shows the monitoring of the ATM network 10 based on VCC performance monitoring and continuity monitoring at the ATMdown interface of the customer-side network transition unit CP-IWF.","Performance errors or continuity errors on the access digital section DS detected on the line side by the customer-side network transition unit CP-IWF form a substitute for CRC-4 monitoring (error report).","Error monitoring is simplified.","The detection of bit errors in the access digital section DS is possible.","There is no possibility of emulating other cases as specified in the ISDN PRI standard, e.g.","for the case in which an additional Sa6 bit sequence is to be transmitted if a CRC-4 error threshold of 512 is exceeded.","FIG.","5 shows the case in which an errored block consisting of user cells has been detected on the basis of the received FPM cells or a loss of continuity (LOC—Loss of Continuity) has been detected as a result of VCC continuity monitoring.","FIG.","6 shows the monitoring of the ATM network 10 by means of VCC performance monitoring and by means of continuity monitoring at the ATMup interface of the exchange-side network transition unit CO-IWF.","The figure shows the sequences when an error is detected in the ATM signal received by the exchange-side network transition unit CO-IWF.","VCC continuity monitoring is performed according to ITU-T standard I.610.Of the various functions specified in ITU-T standard I.610, the following are used: continuity monitoring (CC—Continuity Check) is activated during the connection setup, the VC CC cells are transmitted repeatedly at a periodicity of nominally one cell per second irrespective of the number of user cells transferred, a loss of continuity (LOC) is detected if the VCC receiver (sink point) receives no user cell or no continuity monitoring cell wihtin a time interval of 3.5 seconds with a margin of 0.5 seconds.","VCC performance monitoring is likewise performed according to ITU-T standard I.610, the following options of ITU-T standard I.610 being used: performance monitoring is activated during the connection establishment, forward performance monitoring is supported, the block size N, after which an FPM cell is inserted, lies outside the regulatory scope of the document, i.e.","it is a configurable parameter.","The function explained with reference to FIG.","5 is also referred to as function F3.The function explained with reference to FIG.","6 is also referred to as function F4.In addition, as shown in FIGS.","7 and 8, a second CRC-4 section is monitored between the customer-side network transition unit CP-IWF and the customer telephony equipment TE.","FIG.","7 shows the function sequences when a CRC-4 error is reported by the customer telephony equipment TE.","The function explained in FIG.","7 is also referred to as function F5.FIG.","8 shows function sequences when a CRC-4 error is detected in the Tup signal of the customer-side network transition unit CP-IWF.","It is the task of the customer-side network transition unit CP-IWF to detect CRC-4 errors and indicate them to the exchange ET.","The associated bit sequences are shown in the table in FIG.","8.The function shown with reference to FIG.","8 is also referred to as function F6.FIG.","9 shows function sequences when a CRC-4 error is reported by the customer telephony equipment TE and a CRC-4 error is simultaneously detected in the Tup signal of the customer-side network transition unit CP-IWF.","The function shown with reference to FIG.","9 is also referred to as function F7.The error condition explained for function F7 is an overlay of the error conditions which have been explained in connection with functions F5 and F6 with reference to FIGS.","7 and 8.Error Indications There follows an overview of the errors that occur and the associated error indications which have to be detected and transmitted in each case by the network transition unit.","These error indications are generated in the network termination unit NT1 or the customer-side network transition unit CP-IWF and transferred in the Sa6 bits to the exchange ET. LOS or LFA at the T Reference Point of the NT1/CP-IWF FIG.","10 shows function sequences when a loss of signal (LOS) or loss of frame alignment (LFA) is detected at the Tup interface of the customer-side network transition unit CP-IWF.","When the loss of signal (LOS) or loss of frame alignment (LFA) is detected, the customer-side network transition unit CP-IWF transmits the Sa6 bit sequence with the value “1100” in the transmission direction toward the exchange-side network transition unit CO-IWF.","The exchange-side network transition unit CO-IWF transmits the received Sa6 bit sequence unchanged, i.e.","transparently, to the exchange ET. After receiving the Sa6 bit sequence, the exchange ET sets the A bit to the value one and the Sa4 bit to the value zero.","These bits are inserted in time slot TS0 in the direction toward the customer.","In addition the customer-side network transition unit CP-IWF inserts the E bit containing the value zero into the signal directed toward the customer telephony equipment TE or into the downstream signal.","The function explained with reference to FIG.","10 is also referred to as function F8.LOS or LFA on the Line Side of the NT1/CP-IWF FIG.","11 shows function sequences when an ATM connection error is detected at the ATMdown interface of the customer-side network transition unit CP-IWF.","An error in the signal originating from the ATM network 10 is detected in the customer-side network transition unit CP-IWF as a result of end-to-end VCC continuity monitoring, loss of a cell, loss of cell alignment or cell synchronicity or due to a physical connection failure, in particular as a result of a loss of signal (LOS—Loss of Signal) at the XDSL level (XDSL—X-Digital Subscriber Line).","The exchange-side network transition unit CO-IWF begins with VCC continuity monitoring (CC—Continuity Check) after the relevant VCC has been established.","The VCC continuity monitoring is performed according to ITU standard I.610.Of the various options specified in the I.610 standard, the following are used: continuity monitoring (CC) is activated during the connection establishment, the VC CC cells are transmitted repeatedly at a periodicity of nominally one cell per second irrespective of the number of user cells transferred.","The function shown with reference to FIG.","11 is also referred to as function F9.LOS at the V3 Reference Point or at the Exchange-Side Network Transition Unit CO-IWF FIG.","12 shows function sequences when an ATM connection error occurs at the ATMdown interface, the error having been caused by a loss of signal (LOS) at the exchange-side network transition unit CO-IWF.","The function sequences shown in FIG.","12 are similar to the function sequences explained with reference to FIG.","11.The exchange-side network transition unit CO-IWF deactivates the VCC continuity monitoring (CC) for as long as a loss of signal (LOS) is detected.","In addition no user cells, including the cells for transferring the information of time slot TSO, are transmitted.","In this way the customer-side network transition unit CP-IWF can detect an ATM connection error.","The function explained with reference to FIG.","12 is also referred to as function F10.LOS or LFA at the T Reference Point of the NT1/CP-IWF and LOS at the V3 Reference Point of the LT/CO-IWF FIG.","13 shows function sequences when a loss of signal (LOS) or loss of frame alignment (LFA) occurs at the T reference point of the customer-side network transition unit CP-IWF and simultaneously a loss of signal (LOS) is detected at the V3 reference point of the exchange-side network transition unit CO-IWF.","The function explained with reference to FIG.","13 is also referred to as function F11.The error condition explained for function F11 is a combination of the error conditions which have been explained above in connection with the functions F8 and F10 with reference to FIGS.","10 and 12 respectively.","LOS on the Line Side of the LT/CO-IWF FIG.","14 shows function sequences when an ATM connection error occurs in the ATMup signal at the exchange-side network transition unit CO-IWF.","If the exchange-side network transition unit CO-IWF detects an error in the ATMup signal as a result of the non-arrival of VCC continuity monitoring cells, loss of cells, loss of cell alignment or physical connection failure, e.g.","due to a loss of signal (LOS) at XDSL level, the function sequences shown in FIG.","14, which are also referred to as function F12, are executed.","AIS at the V3 Reference Point of the CO-IWF and Transfer to the CP-IWF FIG.","15 shows function sequences when an AIS (Alarm Indication Signal) is detected at the customer-side network transition unit CP-IWF.","The customer-side network transition unit CP-IWF detects the AIS on the line side.","The AIS is generated in the time-division multiplex network 12 and forwarded by the exchange-side network transition unit CO-IWF.","When AIS alarm data packets are received at the ATMdown interface of the customer-side network transition unit CP-IWF, the customer-side network transition unit CP-IWF transmits an AIS to the customer telephony equipment TE, which is also designated by the acronym CPE (Customer Premises Equipment).","The customer telephony equipment TE returns an A bit with the value one, which is transferred unchanged, i.e.","transparently, to the exchange ET.","The cause for the AIS received in the customer-side network transition unit CP-IWF was an AIS bit sequence inserted by the exchange ET.","The function explained with reference to FIG.","15 is also referred to as function F13.LOS/LFA at the Tup Interface of the CP-IWF and AIS at the V3 Reference Point of the CO-IWF FIG.","16 shows function sequences when an AIS is detected in the ATMdown signal and simultaneously a loss of signal (LOS) or loss of frame alignment (LFA) is detected at the T reference point of the customer-side network transition unit CP-IWF.","In this case, the AIS is transferred to the customer-side network transition unit CP-IWF.","The function shown with reference to FIG.","16 is also referred to as function F14.Function 14 is based on an error case which is a combination of two error cases which have been explained above with reference to functions F8 and F13 and also with reference to FIGS.","10 and 15.Operating Voltage Failure in the NT1/CP-IWF FIG.","17 shows function sequences when an operating voltage failure is detected at the customer-side network transition unit CP-IWF.","When an operating voltage failure is detected by the customer-side network transition unit CP-IWF, the bit values shown in the table in FIG.","17 are used.","The function shown in FIG.","17 is also referred to as function F15.Exemplary Embodiment with Specification of the Basic Text for an ATM Standard There follows an explanation of a further exemplary embodiment in which an expanded basic text is proposed for ATM standard af-vmoa-0145.000.Based on the exemplary embodiments already explained above, it is shown which text sections of the af-vmoa-0145.000 standard are to be replaced or supplemented.","New sections are also specified.","Changes or addenda are indicated by a note at the beginning of each section, e.g.","by “replaces section x.y.z of the af-vmoa-0145.000 standard”.","Some sections must be added in the af-vmoa0145.000 standard because the aspects mentioned therein are not covered by the current basic document.","This is indicated by a note, e.g.","by “new section x.y.z required in af-vmoa-0145.000”.","Subsections which contain no such comments are purely informative.","The text proposed in the exemplary embodiment still has to be revised for the purpose of drafting a basic text so as to bring it more closely into line with the structure of the af-vmoa-0145.000 standard.","Objectives (Replaces Section 1.1 of the af-vmoa-0145.000 Standard) The extension of the subscriber line loop emulation service using ARL2 for ISDN PRI services which are described in this document satisfies a need of the market for an efficient transfer method for carrying ISDN PRI traffic over a broadband subscriber line connection such as e.g.","an XDSL line between a customer terminal equipment and the public circuit-switched or line-switched telephone network.","The intention is to use the extension for access trunking to the public circuit-switched network with leased lines, as used mainly for the connection of PBXs (PBX—Private Branch Exchange).","The classical ISDN PRI is based on conventional time-division multiplex (TDM) methods with extended requirements and OAM principles (OAM—Operation Administration Maintenance) which have been specified for monitoring the circuit-switched connection.","For this reason an ISDN PRI emulation service which uses the principles of the ATM/LES must also meet these requirements.","The methods required for this are specified in this document.","Scope (Replaces Section 1.2 of the af-vmoa-0145.000 Standard) This specification explains the methods which permit the efficient transfer of ISDN PRI (2048 kbps/E1 signal) services over an ATM network between two network transition units (IWF—Interworking Function).","The network transition units are located: in the customer area (CP-IWF with the user-side interface at the T reference point, specified in ITU-T standards I.411, I.431 and ETSI ETS 300 011), and in the service provider area (CO-IWF with the local exchange interface at the V3 reference point, specified in ITU-T standard Q.512).","Also specified is the use of virtual ATM links via AAL2 for transporting bearer information and signaling.","The virtual links used shall be PVCs (Permanent Virtual Circuit).","The regulatory scope of this specification covers: the functionality of the network transition units IWF, the relevant aspects of the control level for the subscriber line (loop) emulation service using AAL2 (Loop Emulation Service Using AAL2), the relevant aspects of the user level for the subscriber line (loop) emulation service using AAL2, relevant aspects of the network management level.","Reference Model (Replaces Section 1.4 of the af-vmoa-0145.000 Standard) This specification is intended to support the delivery of ISDN PRI services to customer premises via bandwidth-constrained ATM connections such as those provided for example by DSL (Digital Subscriber Line) systems.","In this reference model, only those entities are shown that pertain to the ISDN PRI services.","In one implementation the equipment providing the CP-IWF function may also include data interfaces toward the customer, e.g.","an Ethernet.","The data traffic originating from such interfaces or terminating at such interfaces is carried over AAL5 or another appropriate AAL on the same ATM interface to the ATM network as the traffic in the voice band.","The service node shown in FIG.","18 stands for an exchange of the public circuit-switched telephone network (PSTN—Public Switched Telephone Network) which provides public telephone switching services and leased line services over a narrowband service node interface (SNI).","The service node can connect the exchange-side network transition unit CO-IWF via one or more physical interfaces.","Alternatively the CO-IWF functionality can be an integral part of the service node with the result that the physical interface between the exchange-side network transition unit CO-IWF and the service node is not visible toward the outside.","The physical connection between the CP-IWF and the ATM network is typically provided by a DSL (Digital Subscriber Line), an HFC (Hybrid Fibre Coax), a fiber optic link or a radio link.","The ATM network may be a full network, a single ATM switching element or simply a direct interconnection between a CO-IWF and a CP-IWF.","The ATM virtual circuits through the ATM network between the CP-IWF and the CO-IWF shall be PVCs or SPVCs (Soft Permanent Virtual Circuits) which carry the following traffic: bearer traffic and signaling using AAL2, where the DSS1 (Digital Signaling System Number One) for controlling the narrowband services is carried in an AAL2 channel within the same ATM VCC as the associated bearer traffic.","The CO-IWF and CP-IWF described in this specification are functional units which may be implemented as standalone devices, as parts of larger devices, or distributed among several different devices.","This specification does not dictate the implementation of any one of these configurations.","CO-IWF Functionality (In Addition to Section 1.7 of the af-vmoa-0145.000 Standard) signaling adaptation (Signaling Interworking) for the transfer of fault messages (failures) and alarm messages (alarms) in order to receive maintenance signaling from the SNI (Service Node Interface) (if present) and from the ATM broadband interfaces and to insert signaling into the SNI and the ATM broadband interfaces.","For example, a loopback command originating from the exchange ET is replaced by a VCC loopback and in the other transmission direction errors received in the AAL2 maintenance channel are mapped to error messages (notifications) via Sa bits into the signal which is not used for frame alignment.","CP-IWF Functionality (In Addition to Section 1.8 of the af-vmoa-0145.000 Standard) signaling adaptation (Signaling Interworking) for the transfer of failures and alarms in order to receive maintenance signaling from the ISDN PRI interfaces or from the ATM broadband interfaces and to insert signaling into the ISDN PRI interfaces or the ATM broadband interfaces.","For example, alarm indications are transferred in the AAL2 maintenance channel.","Interfaces Supported (Replaces Section 2 of the af-vmoa-0145.000 Standard) This specification identifies the narrowband interfaces and specifies the ATM interfaces at the CP-IWF and the CO-IWF.","The IWFs may also provide other interfaces for management and administration purposes, but these are not specified in this document.","IWF Narrowband Interfaces Physical Layer On the physical layer, an IWF (CP- and CO-) should support DS1 or E1 circuits according to the ITU-T standards G.703 and G.704 depending on the application.","The channel structure of the primary multiplex interface (Primary Rate Interface Channel Structure) specified in ITU-T standard I.412 should be supported.","This means: the primary multiplex B channel structure is supported, i.e.","independent B channels operating at 64 kbps.","For the E1 interface operating at 2,048 Mbps, the following applies: 30 B+D.","For the DS1 interface operating at 1,544 Mbps, the following applies: 23 B+D.","The H channel structures are not supported (H0: 384 kbps (kilobits per second), H11: 1536 kbps, H12: 1920 kbps).","IWF User-Side and Network-Side Interfaces A CP-IWF must support suitable interfaces for ISDN PRI connection to a telephony equipment in the customer area.","The specifications for this interface include: a primary multiplex ISDN interface according to the ITU-T I.431 and ETSI ETS 300 011 standards or equivalent national specifications.","A CO-IWF must support appropriate interfaces for ISDN PRI connections to the telephone network (in compliance with ITU-T standard Q.512).","Signaling On the signaling layer, an IWF shall support one of the following signaling systems depending on the required application: Signaling systems with common channel: N-ISDN signaling according to ITU-T standards Q.921 and Q.931 (DSS1).","N-ISDN according to the ETSI version of DSS1, as specified in the ETSI standards ETS 300 125 and ETS ETSI 300 102-1.Capabilities Supported ATM VCCs for Carrying AAL2 Channels for Supporting a PRI Via an LES (Replaces Section 3.2 of the af-vmoa-0145.000 Standard) An ATM VCC between a CP-IWF and a CO-IWF may include traffic from precisely one ISDN primary multiplex interface.","DSS1 signaling for ISDN PRI lines and the B channels of the narrowband interface are carried in the same ATM VCC in different AAL2 channels.","Time slot TS0 of an ISDN PRI shall be mapped into the AAL2 channel with the channel identifier CID=16.Time slot TS0 contains the frame alignment signal or the signal not relating to the frame alignment which is used for transferring the maintenance and alarm information.","Signaling Between CO-IWF and CP-IWF for the ISDN PRI (Replaces Section 3.3.2 of the af-vmoa-0145.000 Standard) The protocol reference model for a CP-IWF with ISDN PRI in the direction toward the user-side interface is illustrated in FIG.","19.The transport of media streams between CP-IWF and CO-IWF is the same as in the case of analog telephony, including the optional use of tone dialing (DTMF Dial Digit Service—Dual Tone Multiple Frequency).","The DSS1 signaling is not terminated in the CP-IWF.","DSS1 signaling messages are relayed by the CP-IWF from the narrowband D channel across the AAL2 VCC using the transmission/error detection service as specified in ITU-T standard I.366.1.Selection and Changing of Coding (The First Part of this Section is Informative) The same methods are valid as specified in the af-vmoa-0145.000 standard because the support for the H channel structures specified in ITU-T standard I.412 is not provided.","These channel structures require time frame integrity.","For this reason, AAL2 methods such as silence suppression and speech compression can only be used simultaneously for the entire H channel.","Thus, for example, a change of coding profile within a profile would only be possible if it simultaneously affects all bearer channels.","This means that in the case of an H0, H11 or H12 channel the profile change must take place for all 6, 23 or 30 B channels simultaneously.","(The Following Text Supplements Section 3.4.2 of the af-vmoa-0145.000 Standard) Time slot TS0 of the ISDN PRI is mapped into the AAL2 channel with the CID=16.The SSCS (Service Specific Convergence Sublayer) specified in ITU-T standard I.366.2 is used in this channel with PCM64 coding and a change of coding profile is not permitted because the information contained in time slot TS0 must be transferred transparently.","Consequently no speech compression and no silence suppression should be used for this channel.","Application Identifier (AppId) (Supplements Section 4.1.1 of the af-vmoa-0145.000 Standard) In addition an application identifier must be specified for: the subscriber line (loop) emulation service using DSS1 to support PRI with ELCP (Emulated Loop Control Protocol).","SSCS Type The SSCS type to be used on each channel of an AAL2 VCC will vary according to the usage of that channel.","Channels that are used to carry media streams (ISDN B channels) shall use the SCCS specified in ITU-T standard I.366.2.The channels that are used to carry the control and management level traffic (ELCP, DSS1 in the ISDN D channels and LES-EOC) shall use the SCCS specified in ITU-T standard I.366.1.The SCCS specified in ITU-T standard ITU-T I.366.2 shall be used to carry additional failure and alarm messages such as are included in time slot TS0 of the ISDN PRI application, but without speech compression and/or silence suppression and VAD methods (Voice Activity Detection).","CID Allocation (Supplements Section 4.4.1 of the af-vmoa-0145.000 Standard) In the case of support for ISDN PRI the CID with the value sixteen shall be used for transferring time slot TS0 of the ISDN PRI signal.","This CID is assigned during system startup, with the channel being activated at the same time.","Signaling and Control Methods The information contained in this section must be assigned to the corresponding sections of the basic text in order to fit into the structure of the af-vmoa-0145.000 standard.","Additional signaling and control methods for ISDN PRI are disclosed.","Signaling Methods for the ISDN PRI The ISDN PRI consists of: 30 time slots, numbered from 1 to 15 and 17 to 31, for transferring the B channels, a signaling time slot with the number 16 for transporting the DSS1 signaling, and a time slot with the number 0 for transferring the time frame alignment of the signal for the CRC multi time frame alignment, the CRC, and maintenance information in the case of an E1 interface.","Each active B channel, the signaling channel (DSS1 signaling), the ELCP protocol and the channel containing the frame alignment signal or the signal not relating to the frame alignment shall be transferred in a separate AAL2 channel of the same VCC.","B channels are allocated dynamically to the AAL2 channels using the ELCP according to the call status.","The AAL2 channels for the ELCP and the frame alignment signal and for the signal not relating to the frame alignment are allocated statically.","For DSS1 signaling, the AAL2 channel is allocated dynamically by the CO-IWF using an ALLOCATION message, as in the case of the ISDN BRI.","For the signal relating to the frame alignment and for the signal not relating to the frame alignment (TS0), the AAL2 channel is identified by the CID with the value sixteen.","Detection of Unused Channels (Idle Channel Detection) The CO-IWF receives no explicit indication of the seizure or release of a channel, as in the case of the LES and a V5 or a GR303 SNI.","The CO-IWF must therefore be informed by means of other measures.","The proposed solution is that the CO-IWF detects the call status “Idle” as a result of the monitoring of the DSS1 messages in time slot sixteen of the narrowband interface.","Based on the messages for call setup and release, the CO-IWF will seize or release the necessary ARL2 channels using ELCP.","The signaling information itself is transferred transparently between the IWFs using the SSSAR (Service Specific Segmentation And Reassembly) and the SSTED methods (Service Specific Transmission Error Detection) in accordance with the af-vmoa-0145.000 standard.","FIG.","20 shows the protocol reference model for the CO-IWF.","ISDN PRI Specific OAM Methods and Tables of the Alarm States (A New Section is Required in the af-vmoa-0145.000 Standard) Loopbacks (The text contained in the Loopbacks section above should be inserted here, i.e.","including the explanations concerning the functions F1 and F2 as well as FIGS.","3 and 4).","Error Monitoring (The text contained in the Error monitoring section above should be inserted here, i.e.","including the explanations concerning the functions F3 to F7 as well as FIGS.","5 to 9).","Error Indication (The text contained in the Error indications section above should be inserted here, i.e.","including the explanations concerning the functions F8 to F15 and FIGS.","10 to 17).","List of Reference Characters DS Access digital section 4, 6 CRC-4 methods TE, CPE Customer telephony equipment, Customer Premises Equipment NT1 Network termination unit LT Line termination unit ET, SN Exchange, Service Node CRC-4 Error code A A bit E E bit Sa4 to Sa8 Sa4 to Sa8 bit CP-IWF Customer-side network transition unit (Customer Premises Interworking Function CO-IWF Exchange-side network transition unit (Central Office Interworking Function 10 ATM network 12 Time-division multiplex network LOS Loss of Signal LFA Loss of Frame Alignment) LOC Loss of Continuity CC Continuity Check) AIS Alarm Indication Signal TS0 Clock start time slot a analyze t forward transparently FPM Forward Performance Monitoring ATM Asynchronous Transfer Mode F1 to F15 Function V3, T Reference point I, II Sub-multi time frame LB Loopback DS1 Connection type (1,544 Mbit/s) E1 Connection type (2,048 Mbit/s) AIS Alarm Indication Signal AUXP Auxiliary Pattern"]],"string":"[\n [\n \"Method for transferring information and associated network transition units\",\n \"Disclosed is a method wherein operative, administrative and maintenance functions are defined initially for a time-division multiplex oriented communication network (12).\",\n \"In order to utilize said functions in a packet-oriented communication network (10), they are emulated by said network (10).\"\n ],\n [\n \"1.A method for transferring information within at least one time-division multiplex oriented communication network via at least one packet-oriented communication network, comprising: implementing functions for operation and/or administration and/or maintenance in the time-division multiplex oriented communication network for transfer of the information within the time-division multiplex oriented communication network; transferring at least a part of the information via the packet-oriented communication network; and emulating at least some of the functions for operation, administration and maintenance of the time-division multiplex oriented communication network by the packet-oriented communication network.\",\n \"2.The method according to claim 1, wherein a customer-side network transition unit is disposed between a customer area and the packet-oriented communication network, and an exchange-side network transition unit is disposed between the packet-oriented communication network and an exchange of the time-division multiplex oriented communication network.\",\n \"3.The method according to claim 2, wherein a function relates to the activation of a loopback from the exchange via the exchange-side network transition unit back to the exchange, the exchange sets at least one Sa5 bit to the value zero and using successive Sa6 bits transmits an Sa6 bit sequence with the value “1111” at least eight times, the values of the Sa5 bits and the Sa6 bit sequence are detected in the exchange-side network transition unit, when the value zero is detected for the Sa5 bit when the value “1111” is detected eight times in succession for the Sa6 bit sequence by the exchange-side network transition unit, the data arriving from the exchange is transmitted back essentially unchanged to the exchange, whereby at least one A bit with the value one and at least one Sa5 bit with the value zero are transmitted from the exchange-side network transition unit to the exchange, and at least one A bit with the value one is transferred by the exchange-side network transition unit via the packet-oriented communication network.\",\n \"4.The method according to claim 2, wherein one of the functions relates to activation of a loopback from the exchange, via the exchange-side network transition unit, via the customer-side network transition unit, back to the exchange-side network transition unit and then back to the exchange, the exchange transmits at least one Sa5 bit with a value zero and an Sa6 bit sequence with a value “1010” at least eight times, the value of the Sa5 bit and the value of the Sa6 bit sequence are detected in the exchange-side network transition unit, when the value zero is detected for the Sa5 bit and when the value “1010” is detected eight times in succession for the Sa6 bit sequence by the exchange-side network transition unit, the data arriving from the exchange is transmitted back essentially unchanged to the exchange, whereby at least one A bit with a value one and at least one Sa5 bit with a value zero are transmitted by the exchange-side network transition unit to the exchange, and whereby the incoming data is manipulated depending on transmission errors in a part of the loopback located in the packet-oriented communication network, the transmission of loopback data packets is initiated by the exchange-side network transition unit, the loopback data packets are detected in the customer-side network transition unit and transmitted back to the exchange-side network transition unit, the exchange-side network transition unit checks the received loopback data packets and depending on the result of the check manipulates the data to be transmitted to the exchange, and upon detection of a loopback data packet at least one A bit with the value one is transmitted to a customer telephony equipment by the customer-side network transition unit.\",\n \"5.The method according to claim 2, wherein one of the functions relates to the monitoring of the performance and continuity of the transmission from the exchange-side network transition unit to the customer-side network transition unit, performance data packets and continuity data packets are generated in the exchange-side network transition unit and transmitted to the customer-side network transition unit, the performance data packets and continuity data packets are monitored in the customer-side network transition unit, when an error is detected by the customer-side network transition unit at least one E bit with a value zero is transmitted via the packet-oriented communication network to the exchange-side network transition unit and is forwarded to the exchange, and in the exchange-side network transition unit a new checksum is computed for the data to be transmitted to the exchange.\",\n \"6.The method according to claim 2, wherein on of the function relates to the monitoring of the performance and continuity of the transmission from the customer-side network transition unit to the exchange-side network transition unit, performance data packets and continuity data packets are generated in the customer-side network transition unit and transmitted to the exchange-side network transition unit, the performance data packets and continuity data packets are monitored in the exchange-side network transition unit, upon an error the computation of a checksum for the data to be transmitted to the exchange is manipulated, and in the error-free case a checksum for the data to be transmitted to the exchange is computed without manipulation.\",\n \"7.The method according to claim 2, wherein one of the function relates to an error which is reported by a customer telephony equipment, that the error monitoring method is performed in the customer telephony equipment, when an error is detected by the customer telephony equipment least one E bit with a value zero is transmitted to the customer-side network transition unit, the value of the E bit is monitored in the customer-side network transition unit, when the value zero is detected for the E bit by the customer-side network transition unit at least one Sa5 bit with the value one and at least one Sa6 bit sequence with the value “0001” are transmitted via the packet-oriented communication network to the exchange-side network transition unit and are forwarded to the exchange.\",\n \"8.The method according to claim 2, wherein one of the function relates to an error which is detected by the customer-side network transition unit, the error monitoring method is performed in the customer-side network transition unit, when an error is detected by the customer-side network transition unit at least one Sa5 bit with a value one and at least one Sa6 bit sequence with a value “0010” are transmitted via the packet-oriented communication network to the exchange-side network transition unit and are forwarded to the exchange, and upon an error being detected by the customer-side network transition unit at least one E bit with the value zero is transmitted to the customer telephony equipment.\",\n \"9.The method according to claim 2, wherein one of the function relates to a first error which is reported by a customer telephony equipment and to a second error which is detected by the customer-side network transition unit, a first error monitoring method is performed in the customer telephony equipment, upon an error being detected by the customer telephony equipment at least one E bit with a value zero is transmitted to the customer-side network transition unit, the values of the E bits are monitored in the customer-side network transition unit, a second error monitoring method is performed in the customer-side network transition unit, and upon an error being detected by the second error monitoring method and when the value zero is detected for the E bit by the customer-side network transition unit at least one Sa5 bit with a value one and at least one Sa6 bit sequence with a value “0011” are transmitted via the packet-oriented communication network to the exchange-side network transition unit and are forwarded to the exchanges.\",\n \"10.The method according to acclaim 2, wherein one of the function relates to the detection of a loss of signal or a loss of frame alignment between a customer telephony equipment and the customer-side network transition unit, upon a loss being detected in the customer-side network transition unit at least one Sa5 bit with a value one and at least one Sa6 bit sequence with a value “1100” are transmitted by the customer-side network transition unit via the packet-oriented communication network to the exchange-side network transition unit and are forwarded to the exchange, and when a loss is detected by the customer-side network transition unit at least one E bit with the value zero is transmitted to the customer telephony equipment.\",\n \"11.The method according to claim 1, wherein one of the functions relates to detection of a loss of continuity or a loss of a cell or a loss of cell alignment or a physical connection failure in the packet-oriented communication network, continuity data packets are generated in the exchange-side network transition unit and transmitted to the customer-side network transition unit, the continuity data packets are detected in the customer-side network transition unit, upon non-arrival of the continuity data packets or upon a loss of cell or a loss of cell alignment or a physical connection failure being detected by the customer-side network transition unit, a continuous bit sequence of bits with a value one is transmitted to a customer telephony equipment, and upon non-arrival of the continuity data packets or upon a loss of cell or a loss of cell alignment or a physical connection failure being detected in the customer-side network transition unit A bits originating from the customer telephony equipment are transmitted unchanged via the packet-oriented communication network, and upon non-arrival of the continuity data packets or upon a loss of cell or a loss of cell alignment or a physical connection failure being detected by the customer-side network transition unit at least one Sa5 bit with a value one and at least one Sa6 bit sequence with a value “1110” are transmitted via the packet-oriented communication network to the exchange-side network transition unit and are forwarded to the exchange.\",\n \"12.The method according to acclaim 2, wherein one of the functions relates to detection of a loss of signal between the exchange and the exchange-side network transition unit, upon a loss being detected in the exchange-side network transition unit by the exchange-side network transition unit no further continuity data packets are transmitted via the packet-oriented communication network, the continuity data packets are monitored in the customer-side network transition unit, upon non-arrival of the continuity data packets a continuous bit sequence of bits with a value one is transmitted by the customer-side network transition unit to a customer telephony equipment, upon non-arrival of the continuity data packets in the customer-side network transition unit A bits originating from the customer telephony equipment are transmitted unchanged via the packet-oriented communication network, and upon non-arrival of the continuity data packets from the customer-side network transition unit at least one Sa5 bit with a value one and at least one Sa6 bit sequence with a value “1110” are transmitted via the packet-oriented communication network to the exchange-side network transition unit and are forwarded to the exchange.\",\n \"13.The method according to claim 1, wherein one of the functions relates to detection of a loss of signal or a loss of frame alignment between a customer telephony equipment and the customer-side network transition unit and to detection of a loss of signal between the exchange and the exchange-side network transition unit, upon a loss of signal being detected between the exchange and the exchange-side network transition unit in the exchange-side network transition unit no further continuity data packets are transmitted via the packet-oriented communication network, the continuity data packets are monitored in the customer-side network transition unit, upon non-arrival of the continuity data packets a continuous bit sequence of bits with a value one is transmitted by the customer-side network transition unit to the customer telephony equipment, upon non-arrival of the continuity data packets at least one Sa5 bit with a value one and at least one Sa6 bit sequence with a value “1110” are transmitted by the customer-side network transition unit via the packet-oriented communication network to the exchange-side network transition unit and from there are forwarded to the exchange, and upon a loss of signal or a loss of frame alignment being detected between the customer telephony equipment and the customer-side network transition unit and simultaneous non-arrival of the continuity data packets at least one A bit with a value zero is transmitted by the customer-side network transition unit via the packet-oriented communication network to the exchange-side network transition unit and is forwarded to the exchange.\",\n \"14.The method according to claim 2, wherein one of the function relates to detection of a loss of continuity or a loss of cell or a loss of cell alignment or a physical connection failure between the customer-side network transition unit and the exchange-side network transition unit, continuity data packets are generated in the customer-side network transition unit and transmitted to the exchange-side network transition unit, upon non-arrival of the continuity data packets or upon a loss of cell or a loss of cell alignment or a physical connection failure being detected by the exchange-side network transition unit a bit sequence conforming to a predefined auxiliary bit pattern is transmitted to the exchange, and the exchange, after receiving the bit sequence, transmits an A bit with the value one to the exchange-side network transition unit.\",\n \"15.The method according to acclaim 2, wherein one of the function relates to detection of a continuous bit sequence of bits with a value one, an occurrence of the continuous bit sequence of bits with a value one originating from the exchange is monitored in the exchange-side network transition unit, that when the bit sequence is detected at least one alarm data packet is generated and transmitted via the packet-oriented communication network to the customer-side network transition unit, the occurrence of alarm data packets is detected in the customer-side network transition unit, when an alarm data packet is detected by the customer-side network transition unit the continuous bit sequence of bits with a value one is transmitted to a customer telephony equipment, when an alarm data packet is detected in the customer-side network transition unit A bits originating from the customer telephony equipment are transmitted unchanged via the packet-oriented communication network, and when an alarm data packet is detected by the customer-side network transition unit at least one Sa6 bit sequence with a value “1111” is transmitted via the packet-oriented communication network to the exchange-side network transition unit and is forwarded to the exchange.\",\n \"16.The method according to claim 2, wherein on of the functions relates to detection of a continuous bit stream of bits with a value one and to detection of a loss of signal or a loss of frame alignment between a customer telephony equipment and the customer-side network transition unit, an occurrence of the continuous bit sequence of bits with a value one originating from the exchange is monitored in the exchange-side network transition unit, when the bit sequence is detected at least one alarm data packet is generated and transmitted via the packet-oriented communication network to the customer-side network transition unit, the occurrence of alarm data packets and the signal and the frame alignment between the customer telephony equipments and the customer-side network transition unit are monitored in the customer-side network transition unit, when an alarm data packet tis detected by the customer-side network transition unit the continuous bit sequence of bits with a value one is transmitted to the customer telephony equipment, when an alarm data packet and a loss of signal or a loss of frame alignment is detected, at least one A bit with a value zero is transmitted by the customer-side network transition unit via the packet-oriented data communication network to the exchange-side network transition unit and from there is forwarded to the exchanges, and when an alarm data packet is detected by the customer-side network transition unit at least one Sa5 bit with a value one and at least one Sa6 bit sequence with a value “1111” are transmitted via the packet-oriented communication network to the exchange-side network transition unit and are forwarded to the exchange.\",\n \"17.The method according to claim 2, wherein one of the functions relates to detection of a voltage failure in the customer-side network transition unit, the operating voltage is monitored in the customer-side network transition unit, upon failure of the operating voltage in the customer-side network transition unit at least one Sa6 bit sequence with a value “1000” is transmitted via the packet-oriented communication network to the exchange-side network transition unit and is forwarded to the exchange, upon failure of the operating voltage in the customer-side network transition unit at least one E bit with a value zero is transmitted to a customer telephony equipment, and the exchange-side network transition unit transmits the Sa6 bit sequence unchanged and subsequently a bit sequence conforming to a predefined auxiliary bit pattern to the exchange.\",\n \"18.The method according to claim 2, wherein the emulation is implemented by functions of the packet-oriented communication network.\",\n \"19.The method according to claim 2, wherein the packet-oriented communication network is implemented according to asynchronous transfer mode.\",\n \"20.The method according to claim 2, wherein the packet-oriented communication network is implemented according to the asynchronous transfer mode Adaption Layer 1 or according to the asynchronous transfer mode Adaption Layer 2.21.The method according to claim 2, wherein the packet-oriented communication network is implemented according to the Internet Protocol.\",\n \"22.The method according to claim 2, wherein the time-division multiplex oriented communication network is an ISDN network.\",\n \"23.The method according to claim 22, wherein part of a primary multiplex access is replaced by the packet-oriented communication network, and/or the primary multiplex access includes an XDSL line between the customer-side network transition unit and the packet-oriented communication network.\",\n \"24.The method according to claim 2, wherein operation, administration and maintenance functions are provided essentially according to ETSI standard ETS 300 011 and/or according to ITU-T standard G.962 and/or according to ETSI standard ETS 300 233.25.The method according to claim 2, wherein a multi time frame including sixteen time frames according to ITU-T standard G.704 is used in the time-division multiplex oriented communication network, the multi time frame including an A bit in a third position and/or an Sa5 bit in a fifth position and/or an Sa6 bit in a sixth position in a start time slot of every second time frame beginning with the second start time slot of the multi time frame, and/or including an E bit in the first position of the second-to-last time frame and/or the last time frame of a multi time frame.\",\n \"26.The method according to claim 25, wherein the data of the start time slot of each time frame is transmitted unchanged in value via the packet-oriented communication network.\",\n \"27.The method according claim 2, wherein located between a customer telephony equipment and the time-division multiplex oriented communication network a broadband subscriber line via which primary multiplex traffic is carried.\",\n \"28.\",\n \"(Canceled) 29.\",\n \"(Canceled)\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>The time-division multiplex oriented communication network is, for example, an ISDN data transmission network (ISDN—Integrated Services Digital Network).\",\n \"In the time-division multiplex oriented communication network the data is transferred in different time slots in accordance with a time-division multiplex method.\",\n \"The packet-oriented communication network is a network in which the information or data is transferred in data packets.\",\n \"The packet-oriented communication network is for example a communication network operating according to the Internet Protocol.\",\n \"Another example of a packet-oriented communication network is an ATM network (ATM—Asynchronous Transfer Mode), in which, however, the data packets are referred to as cells.\",\n \"Functions for operation, administration and/or maintenance in the time-division multiplex oriented communication network have been defined for example in the following standards of the ETSI (European Telecommunications Standards Institute) or ITU-T (International Telecommunication Union—Telecommunication Standardization Sector): ETSI ETS 300 233, Integrated Services Digital Network (ISDN); Access Digital Section for ISDN Primary Rate, May 1994, ITU-T G.962, Digital Sections and Digital Line Systems; Access Digital Section for ISDN Primary Rate at 2048 kbit/s, March 1993, ETSI ETS 300 011, Integrated Services Digital Network (ISDN); Primary Rate User-Network Interface Layer 1 Specification and Test Principles, April 1992.Thus, the functions for operation, administration and/or maintenance relate for example to the activation of loopbacks or to error monitoring.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>The invention relates to a method for transferring information within at least one time-division multiplex oriented communication network via at least one packet-oriented communication network.\",\n \"Functions for operation and/or administration and/or maintenance are implemented in the time-division multiplex oriented communication network with the aim of transferring information within the time-division multiplex oriented communication network.\",\n \"The information relates for example to user data or voice data.\",\n \"The invention discloses, in one embodiment, a method for transmitting information within at least one time-division multiplex oriented communication network via at least one packet-oriented communication network, the method continuing to allow functions for operation, administration and/or maintenance of the time-division multiplex oriented communication network to be used in the time-division multiplex oriented communication network.\",\n \"Associated network transition units are also to be specified.\",\n \"In another embodiment according to the invention, at least some of the information is transferred via the packet-oriented communication network.\",\n \"At the same time at least some of the functions for operation, administration and/or maintenance are emulated by the packet-oriented communication network.\",\n \"As a result the functions specified for the time-division multiplex oriented communication network can continue to be used essentially without restriction.\",\n \"In another embodiment according to the invention, a customer-side network transition unit is interposed between a customer area and the packet-oriented communication network.\",\n \"The customer-side network transition unit is also referred to as a CP-IWF (Customer Premises—Interworking Function).\",\n \"In the embodiment, an exchange-side network transition unit is additionally interposed between the packet-oriented communication network and an exchange of the time-division multiplex oriented communication network.\",\n \"The exchange-side network transition unit is also referred to as a CO-IWF (Central Office—Interworking Function).\",\n \"In other embodiments according to the invention individual functions for operation, administration and maintenance are.\",\n \"In this case, the functions known from the time-division multiplex oriented communication network are retained in the time-division multiplex oriented communication network and emulated in the packet-oriented communication network.\",\n \"In still another embodiment according to the invention, the emulation is provided by functions of the packet-oriented communication network, in particular by the customer-side network transition unit and by the exchange-side network transition unit.\",\n \"In another embodiment, the packet-oriented communication network is an ATM network which is implemented according to the asynchronous transfer mode (ATM).\",\n \"By means of this embodiment powerful methods which have been defined in ATM standards, for example in the standards of the ATM Forum, can be used: af-vtoa-0113.000, ATM Trunking Using AAL-2 for Narrowband Services, February 1999, af-vmoa-0145.000, Voice and Multimedia over ATM—Loop Emulation Service Using AAL-2, July 2000.In another embodiment, the packet-oriented communication network is implemented according to the ATM Adaption Layer 1 or according to the ATM Adaption Layer 2.These layers are also referred to as AAL1 and AAL2 respectively (AAL—ATM Adaption Layer).\",\n \"The AAL2 layer in particular is well suited to a reduction in bandwidth during the transmission in the packet-oriented communication network, in particular for the connection of subscribers.\",\n \"Reference is made for example to the standard af-vmoa-0145.000, Section 2.3, CP-IWF ATM Interfaces, 2.3.1 Physical Layer, where XDSL methods (XDSL—X-Digital Subscriber Line) are cited as examples of transmission methods, i.e.\",\n \"the ADSL method (ADSL—Asymmetrical Digital Subscriber Line) and the SDSL method (SDSL—Symmetrical Digital Subscriber Line).\",\n \"In yet another embodiment the packet-oriented communication network is implemented according to the Internet Protocol.\",\n \"In this way, for example, the IP over ATM transmission method can be used.\",\n \"In another embodiment, the time-division multiplex oriented communication network is an ISDN network.\",\n \"In connection with the invention the following standards relating to the ISDN network are particularly relevant: ITU-T I.411, ISDN User Network Interfaces—Reference Configuration, ITU-T I.412, ISDN User Network Interfaces—Interface Structures and Access Capabilities, and ITU-T Q.512, Exchange Interfaces for Subscriber Access.\",\n \"In another embodiment, parts of a primary multiplex access are replaced by the packet-oriented communication network.\",\n \"The primary multiplex access is also referred to as a Primary Rate Access.\",\n \"In one embodiment, the primary multiplex access is a so-called E1 access with a transmission capacity of 2,048 Mbit/s or a so-called DS1 access with a transmission capacity of 1,544 Mbit/s.\",\n \"The three above-mentioned standards relate to an E1 access.\",\n \"Integrating the packet-oriented communication network with the primary multiplex access allows the transmission bandwidth required on a subscriber access line to be considerably reduced.\",\n \"This enables transmission over two-wire copper lines, for example according to an XDSL method.\",\n \"In one embodiment, furthermore, a multi time frame consisting of sixteen time frames, as described for example in ITU-T standard G.704, is used in the time-division multiplex oriented communication network.\",\n \"This time frame is explained in more detail below with reference to FIG.\",\n \"2 .\",\n \"The time frame includes in particular a so-called A bit for indicating alarm conditions, a so-called Sa5 bit, a so-called Sa6 bit and a so-called E bit for indicating error conditions.\",\n \"The bits are transferred in the start time slots TSO of each time frame.\",\n \"In another embodiment, the start time slot TSO of each time frame is transferred over the packet-oriented communication network unchanged except for the mentioned changes in value, i.e.\",\n \"in particular without speech compression and without silence suppression.\",\n \"Another embodiment relates to a customer-side network transition unit and to an exchange-side network transition unit, each of which contain function units during the operation of which the method steps related to the customer-side network transition unit and to the exchange-side network transition unit are performed according to the invention or one of its embodiments.\",\n \"Thus, the above-mentioned technical effects also apply to the two network transition units.\"\n ],\n [\n \"CLAIM FOR PRIORITY This application claims priority to International Application No.\",\n \"PCT/EP02/03753, which was published in the German language on Oct. 17, 2002, which claims the benefit of priority to German Application No.\",\n \"01108481.1 which was filed in the German language on Apr.\",\n \"4, 2001.TECHNICAL FIELD OF THE INVENTION The invention relates to a method for transferring information and associated network transition units, and in particular, a method for transferring information within at least one time-division multiplex oriented communication network via at least one packet-oriented communication network.\",\n \"BACKGROUND OF THE INVENTION The time-division multiplex oriented communication network is, for example, an ISDN data transmission network (ISDN—Integrated Services Digital Network).\",\n \"In the time-division multiplex oriented communication network the data is transferred in different time slots in accordance with a time-division multiplex method.\",\n \"The packet-oriented communication network is a network in which the information or data is transferred in data packets.\",\n \"The packet-oriented communication network is for example a communication network operating according to the Internet Protocol.\",\n \"Another example of a packet-oriented communication network is an ATM network (ATM—Asynchronous Transfer Mode), in which, however, the data packets are referred to as cells.\",\n \"Functions for operation, administration and/or maintenance in the time-division multiplex oriented communication network have been defined for example in the following standards of the ETSI (European Telecommunications Standards Institute) or ITU-T (International Telecommunication Union—Telecommunication Standardization Sector): ETSI ETS 300 233, Integrated Services Digital Network (ISDN); Access Digital Section for ISDN Primary Rate, May 1994, ITU-T G.962, Digital Sections and Digital Line Systems; Access Digital Section for ISDN Primary Rate at 2048 kbit/s, March 1993, ETSI ETS 300 011, Integrated Services Digital Network (ISDN); Primary Rate User-Network Interface Layer 1 Specification and Test Principles, April 1992.Thus, the functions for operation, administration and/or maintenance relate for example to the activation of loopbacks or to error monitoring.\",\n \"SUMMARY OF THE INVENTION The invention relates to a method for transferring information within at least one time-division multiplex oriented communication network via at least one packet-oriented communication network.\",\n \"Functions for operation and/or administration and/or maintenance are implemented in the time-division multiplex oriented communication network with the aim of transferring information within the time-division multiplex oriented communication network.\",\n \"The information relates for example to user data or voice data.\",\n \"The invention discloses, in one embodiment, a method for transmitting information within at least one time-division multiplex oriented communication network via at least one packet-oriented communication network, the method continuing to allow functions for operation, administration and/or maintenance of the time-division multiplex oriented communication network to be used in the time-division multiplex oriented communication network.\",\n \"Associated network transition units are also to be specified.\",\n \"In another embodiment according to the invention, at least some of the information is transferred via the packet-oriented communication network.\",\n \"At the same time at least some of the functions for operation, administration and/or maintenance are emulated by the packet-oriented communication network.\",\n \"As a result the functions specified for the time-division multiplex oriented communication network can continue to be used essentially without restriction.\",\n \"In another embodiment according to the invention, a customer-side network transition unit is interposed between a customer area and the packet-oriented communication network.\",\n \"The customer-side network transition unit is also referred to as a CP-IWF (Customer Premises—Interworking Function).\",\n \"In the embodiment, an exchange-side network transition unit is additionally interposed between the packet-oriented communication network and an exchange of the time-division multiplex oriented communication network.\",\n \"The exchange-side network transition unit is also referred to as a CO-IWF (Central Office—Interworking Function).\",\n \"In other embodiments according to the invention individual functions for operation, administration and maintenance are.\",\n \"In this case, the functions known from the time-division multiplex oriented communication network are retained in the time-division multiplex oriented communication network and emulated in the packet-oriented communication network.\",\n \"In still another embodiment according to the invention, the emulation is provided by functions of the packet-oriented communication network, in particular by the customer-side network transition unit and by the exchange-side network transition unit.\",\n \"In another embodiment, the packet-oriented communication network is an ATM network which is implemented according to the asynchronous transfer mode (ATM).\",\n \"By means of this embodiment powerful methods which have been defined in ATM standards, for example in the standards of the ATM Forum, can be used: af-vtoa-0113.000, ATM Trunking Using AAL-2 for Narrowband Services, February 1999, af-vmoa-0145.000, Voice and Multimedia over ATM—Loop Emulation Service Using AAL-2, July 2000.In another embodiment, the packet-oriented communication network is implemented according to the ATM Adaption Layer 1 or according to the ATM Adaption Layer 2.These layers are also referred to as AAL1 and AAL2 respectively (AAL—ATM Adaption Layer).\",\n \"The AAL2 layer in particular is well suited to a reduction in bandwidth during the transmission in the packet-oriented communication network, in particular for the connection of subscribers.\",\n \"Reference is made for example to the standard af-vmoa-0145.000, Section 2.3, CP-IWF ATM Interfaces, 2.3.1 Physical Layer, where XDSL methods (XDSL—X-Digital Subscriber Line) are cited as examples of transmission methods, i.e.\",\n \"the ADSL method (ADSL—Asymmetrical Digital Subscriber Line) and the SDSL method (SDSL—Symmetrical Digital Subscriber Line).\",\n \"In yet another embodiment the packet-oriented communication network is implemented according to the Internet Protocol.\",\n \"In this way, for example, the IP over ATM transmission method can be used.\",\n \"In another embodiment, the time-division multiplex oriented communication network is an ISDN network.\",\n \"In connection with the invention the following standards relating to the ISDN network are particularly relevant: ITU-T I.411, ISDN User Network Interfaces—Reference Configuration, ITU-T I.412, ISDN User Network Interfaces—Interface Structures and Access Capabilities, and ITU-T Q.512, Exchange Interfaces for Subscriber Access.\",\n \"In another embodiment, parts of a primary multiplex access are replaced by the packet-oriented communication network.\",\n \"The primary multiplex access is also referred to as a Primary Rate Access.\",\n \"In one embodiment, the primary multiplex access is a so-called E1 access with a transmission capacity of 2,048 Mbit/s or a so-called DS1 access with a transmission capacity of 1,544 Mbit/s.\",\n \"The three above-mentioned standards relate to an E1 access.\",\n \"Integrating the packet-oriented communication network with the primary multiplex access allows the transmission bandwidth required on a subscriber access line to be considerably reduced.\",\n \"This enables transmission over two-wire copper lines, for example according to an XDSL method.\",\n \"In one embodiment, furthermore, a multi time frame consisting of sixteen time frames, as described for example in ITU-T standard G.704, is used in the time-division multiplex oriented communication network.\",\n \"This time frame is explained in more detail below with reference to FIG.\",\n \"2.The time frame includes in particular a so-called A bit for indicating alarm conditions, a so-called Sa5 bit, a so-called Sa6 bit and a so-called E bit for indicating error conditions.\",\n \"The bits are transferred in the start time slots TSO of each time frame.\",\n \"In another embodiment, the start time slot TSO of each time frame is transferred over the packet-oriented communication network unchanged except for the mentioned changes in value, i.e.\",\n \"in particular without speech compression and without silence suppression.\",\n \"Another embodiment relates to a customer-side network transition unit and to an exchange-side network transition unit, each of which contain function units during the operation of which the method steps related to the customer-side network transition unit and to the exchange-side network transition unit are performed according to the invention or one of its embodiments.\",\n \"Thus, the above-mentioned technical effects also apply to the two network transition units.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS Exemplary embodiments of the invention are explained in the following with reference to the attached drawings, in which: FIG.\",\n \"1 shows an ISDN primary multiplex interface model with processing of a time slot TSO according to ETSI standard ETS 300 233.FIG.\",\n \"2 shows a CRC-4 multi time frame structure.\",\n \"FIG.\",\n \"3 shows function units and function sequences for activating a loopback in an exchange-side network transition unit CO-IWF.\",\n \"FIG.\",\n \"4 shows function units and function sequences for activating a loopback in a customer-side network transition unit CP-IWF.\",\n \"FIG.\",\n \"5 shows the monitoring of an ATM network by means of VCC (Virtual Channel Connection) performance monitoring (PMo—Performance Monitoring) and continuity monitoring (CC—Continuity Check) at an ATMdown interface of the customer-side network transition unit CP-IWF.\",\n \"FIG.\",\n \"6 shows the monitoring of the ATM network by means of VCC performance monitoring and continuity monitoring at an ATMup interface of the exchange-side network transition unit CO-IWF.\",\n \"FIG.\",\n \"7 shows function sequences when a CRC-4 error (Cyclic Redundancy Check) is reported by a customer telephony equipment TE.\",\n \"FIG.\",\n \"8 shows function sequences when a CRC-4 error is detected in a Tup signal of the customer-side network transition unit CP-IWF.\",\n \"FIG.\",\n \"9 shows function sequences when a CRC-4 error is reported by the customer equipment TE and simultaneously a CRC-4 error is detected in the Tup signal of the customer-side network transition unit CP-IWF.\",\n \"FIG.\",\n \"10 shows function sequences when a loss of signal (LOS—Loss of Signal) is detected or when a loss of frame alignment or frame synchronization (LFA—Loss of Frame Alignment) is detected at the Tup interface of the customer-side network transition unit CP-IWF.\",\n \"FIG.\",\n \"11 shows function sequences when an ATM connection error is detected at the ATMdown interface of the customer-side network transition unit CP-IWF.\",\n \"FIG.\",\n \"12 shows function sequences when an ATM connection error occurs at the ATMdown interface as a result of a loss of signal (LOS) at the exchange-side network transition unit CO-IWF.\",\n \"FIG.\",\n \"13 shows function sequences when a loss of signal (LOS) or loss of frame alignment (LFA) occurs at the T reference point of the customer-side network transition unit CP-IWF and simultaneously a loss of signal (LOS) occurs at the V3 reference point of the exchange-side network transition unit CO-IWF.\",\n \"FIG.\",\n \"14 shows function sequences when an ATM connection error occurs in the ATMup signal at the exchange-side network transition unit CO-IWF.\",\n \"FIG.\",\n \"15 shows function sequences when an AIS (Alarm Indication Signal) is detected at the customer-side network transition unit CP-IWF.\",\n \"FIG.\",\n \"16 shows function sequences when an AIS is detected in the ATMdown signal and simultaneously a loss of signal (LOS) or loss of frame alignment (LFA) occurs at the T reference point of the customer-side network transition unit CP-IWF.\",\n \"FIG.\",\n \"17 shows function sequences when an operating voltage failure is detected in the customer-side network transition unit CP-IWF.\",\n \"FIG.\",\n \"18 shows the reference model for the subscriber line (loop) emulation service using AAL2.FIG.\",\n \"19 shows a protocol reference model for a customer-side network transition unit CP-IWF with a user-side ISDN PRI and with DSS1 forwarding (DSS1—Digital Signaling System Number One) via the AAL2 between the network transition units IWF.\",\n \"FIG.\",\n \"20 shows a protocol reference model for an exchange-side network transition unit CO-IWF with user-side ISDN PRI and with DSS1 forwarding as well as with L3 (Layer 3) monitoring via the AAL2 between the network transition units IWF.\",\n \"DETAILED DESCRIPTION OF THE INVENTION Introduction to the Technical Environment There are many reasons why support for an ISDN primary multiplex interface via the AAL2 (ATM Adaption Layer) using the principles and methods of the subscriber line emulation services (LES—Loop Emulation Services) according to the af-vmoa-0145.000 standard is desirable.\",\n \"A concept for supporting an ISDN primary multiplex interface via AAL2/LES (ATM Adaption Layer 2/Loop Emulation Service) is disclosed in the following.\",\n \"Also, a text for supplementing the subscriber line emulation service (LES—Loop Emulation Service) using the AAL2 is proposed according to an exemplary embodiment based on the af-vxnoa-0145.000 standard, wherein the ISDN primary multiplex interface (PRI—Primary Rate Interface) is defined as a user-side interface.\",\n \"At the same time consistency with the support for the ISDN basic access interface (BRI—Basic Rate Interface) via AAL2 is ensured, as explained in the af-vmoa-0145.000 standard.\",\n \"The aim is to specify a solution that advantageously uses the powerful methods which have been defined for the subscriber line (loop) emulation service and which to date are not to be found in the approach for channel group transmission via AAL2 (AAL2 trunking).\",\n \"These powerful mechanisms include support for an embedded operation channel (EOC—Embedded Operation Channel), flexible assignment of a channel identifier (CID—AAL2 Channel Identifier) as well as channel activation using the ELCP protocol (ELCP—Emulated Loop Control Protocol).\",\n \"ISDN PRI Overview An overview of the use of the ISDN primary multiplex interface in the world of classical time-division multiplex (TDM—Time Division Multiplex) is given below.\",\n \"Also specified are requirements which must be met if parts of the access digital section (DS—Digital Section) of the primary multiplex access are replaced by the ATM network.\",\n \"Furthermore, methods which meet these requirements are specified.\",\n \"There are differences between the so-called DS1 (Digital Signal Level Number One) and E1 structures (E1—European Digital Signal Level Number One).\",\n \"The exemplary embodiments and explanations relate to the E1 structure.\",\n \"General statements applicable to both interfaces are however made wherever possible.\",\n \"ISDN Primary Multiplex Interface Model (PRI) with Operating and Maintenance Functions (for a 2048 kbps signal/E1) FIG.\",\n \"1 shows an access digital section DS with its delimitations and the processing of time slot TS0.The operating and maintenance functions support methods and information elements which are required for control of the access digital section by an exchange ET or a service node.\",\n \"An Sa5, Sa6, E and A bit of a time slot TS0 are relevant for indication and control purposes.\",\n \"The bit structure of the time slot TS0 and the multi time frame structure are defined according to ITU-T standard G.704 and are explained in more detail below with reference to FIG.\",\n \"2.The A bit is used for passing alarm status information between the service node and a customer telephony equipment TE (customer telephony end equipment).\",\n \"The A bit requires monitoring and is transferred transparently.\",\n \"Other control bits of the time slot TSO are to be transferred transparently.\",\n \"Also shown in FIG.\",\n \"1 are the elements of the access digital section DS which use CRC methods 4, 6 (CRC—Cyclic Redundancy Check).\",\n \"The CRC-4 methods 4, 6 are implemented and used between the exchange ET and the network termination unit NT1 as well as between the network termination unit NT1 and the customer telephony equipment TE.\",\n \"This is also known as Option Two according to ITU-T standard G.962.FIG.\",\n \"1 also shows a line termination unit LT.\",\n \"Between the line termination unit LT and the exchange ET there is a V3 reference point.\",\n \"Between the customer telephony equipment TE and the network termination unit NT1 there is a T reference point.\",\n \"The following table shows the signals which are exchanged between the T reference point and the access digital section DS during the normal operating conditions and error conditions specified in ETSI standard ETS 300 011: Name List of signals Normal operating mode Operating time frame with: time frame active assigned CRC bits, CRC error information (cf.\",\n \"ITU-T standard G.704 for 2048 Kbit/s systems), no error indications RAI (Remote Alarm Indication) Operating time frame with: active assigned CRC bits, CRC error information (cf.\",\n \"ITU-T standard G.704 for 2048 Kbit/s systems), remote alarm indication (cf.\",\n \"ITU-T standard G.704, Table 4a, for 2048 Kbit/s systems, A bit is set to the value one) LOS (Loss of Signal) No input signal received (LOS) AIS (Alarm Indication Signal) Continuous bit stream of bits with the value one CRC error information E bit according to ITU-T standard G.704, Table 4b, set to the value zero if an errored CRC block is received (for a 2048 Kbit/s system only) The signals exchanged between the access digital section DS and the exchange ET are specified in the following table: Name List of signals Normal operating mode Operating time frame with: time frame active assigned CRC bits, CRC error information (cf.\",\n \"ITU-T standard G.704 for 2048 Kbit/s systems), no error indicators RAI (Remote Alarm Indication) Operating time frame with: active assigned CRC bits, CRC error information (cf.\",\n \"ITU-T standard G.704 for 2048 Kbit/s systems), remote alarm indication (cf.\",\n \"ITU-T standard G.704, Table 4a, for 2048 Kbit/s systems, A bit is set to the value one) LOS (Loss of Signal) No input signal received (LOS) AIS (Alarm Indication Signal) Continuous bit stream of bits with the value one CRC error information E bit according to ITU-T standard G.704, Table 4b, set to the value zero if an errored CRC block is received (for a 2048 Kbit/s system only) The following additional signals are required in order to indicate error conditions which occur in relation to the access digital section DS: Name List of signals Normal time frames These are time frames without error indications or loopback requests generated by the exchange ET or the customer telephony equipment TE, where an A bit with the value one or zero is not relevant to the access digital section DS.\",\n \"Time frames These are time frames which, in addition to the normal time frames, have Sa6 bits containing error indication signals which have been generated in the network termination unit NT1 and transmitted to the exchange ET. Alternatively, the Sa6 bits may contain loopback requests which have been transmitted from the exchange ET to the access digital section DS.\",\n \"In this case the Sa5 bit is also used to indicate the direction and as a loopback indication.\",\n \"The Sa5 bit in the transmission direction from the access digital section DS to the exchange ET is set in the network termination unit NT1 or in the line termination unit LT and transmitted to the exchange ET according to the following rules: Sa5 = 1 Loopback 2 not activated, Sa5 = 0 Loopback 2 activated.\",\n \"The Sa6 bits are numbered Sa6(1), Sa6(2), Sa6(3), Sa6(4) and synchronized with the sub- multi time frames which are explained in more detail below with reference to FIG.\",\n \"3.Substitute time frame In the event of a loss of signal (LOS) or loss of frame alignment (LFA) at the T reference point of the network termination unit NT1, a new time frame must be generated.\",\n \"The A bit is set to the value zero and the Sa4, Sa5, Sa7 and Sa8 bits as well as the bits of the time slots TS1 to TS31 are set to the value one.\",\n \"A bit sequence of Sa6 bits is used to indicate this error condition.\",\n \"LFA Loss of Frame Alignment Operating voltage failure in the network termination unit NT1 or the line termination unit LT Auxiliary bit pattern This is a time frame less and (AUXP—Auxiliary Pattern) continuous bit sequence of bits with alternating values of one and zero ( .\",\n \".\",\n \".\",\n \"101010 .\",\n \".\",\n \". )\",\n \"which is transmitted in both transmission directions by the line termination unit LT if a loss of signal (LOS) is detected at the corresponding receive unit FIG.\",\n \"2 shows the structure of a CRC-4 multi time frame.\",\n \"The CRC-4 multi time frame consists of two sub-multi time frames I and II.\",\n \"Each sub-multi time frame I, II consists of eight time frames.\",\n \"The CRC-4 checksum is computed for all the bits of a sub-multi time frame I, II.\",\n \"The CRC bits C1, C2, C3 and C4 are transferred in bit one of the frame alignment signal.\",\n \"The frame alignment signal is transmitted in even-numbered time frames (0, 2, 4, .\",\n \".\",\n \".\",\n \").\",\n \"The Sa bits, like the A bits (Remote Alarm Indication), are part of the signal not relating to the frame alignment.\",\n \"The Sa bits and the A bit are transmitted in the odd-numbered time frames (1, 3, 5, .\",\n \".\",\n \".\",\n \").\",\n \"The first bit in each of the odd-numbered time frames 1, 3, 5, 7, 9 and 11 forms the CRC multi time frame alignment signal, which takes the form of a bit sequence with the value “001011”.\",\n \"Bit one in the thirteenth time frame is the so-called E bit and serves to indicate the reception of an errored sub-multi time frame by setting the E bit from the value one to the value zero for each errored sub-multi time frame I.\",\n \"The first bit in the fifteenth time frame serves to indicate an error for each errored sub-multi time frame II.\",\n \"CRC-4 Method The CRC method serves to provide protection against errored frames (framing) and for error performance monitoring.\",\n \"This includes the multi time frame methods which are specified in ITU-T standard G.704.If the access digital section DS is replaced by the ATM network 10 and the advantages of the AAL2 are used, it is not possible to perform or implement a CRC-4 method between the exchange ET and the network termination unit NT1.For this reason ATM methods are used for this purpose in the exemplary embodiments, although these methods do not offer the same possibilities as CRC monitoring.\",\n \"VCC (Virtual Channel Connection) performance monitoring (PMo—Performance Monitoring) and continuity monitoring (continuity check) are to be used in both transmission directions for monitoring between the network transition unit NT1 and the line termination unit LT. A new computation according to the CRC-4 error monitoring method is performed in the line termination unit LT in the transmission direction toward the exchange ET. No changes are necessary for the CRC-4 method between the network termination unit NT1 and the customer telephony equipment TE.\",\n \"Operation and Maintenance of the Access Digital Section The methods described in the following permit the emulation of an operation and maintenance method for a primary multiplex interface (PRI OAM—Primary Rate Interface Operation Administration Maintenance) via an interface with the subscriber line (loop) emulation service (LES Interface—Loop Emulation Service Interface).\",\n \"The access digital section DS provides the means for transmitting indicator elements and for detecting error conditions at the T reference point interface and the V3 reference point interface as well as for supporting test methods.\",\n \"The following functions are supported according to ETSI standard ETS 300 233: Loopbacks Loopback 1, transparent loopback in the line termination unit LT or in the exchange-side network transition unit CO-IWF (F1), Loopback 2, transparent loopback in the network termination unit NT1 or in the customer-side network transition unit CP-IWF (F2), Error conditions within the access digital section DS loss of signal (LOS) or loss of frame alignment (LFA) on the line side of the network termination unit NT1 or the customer-side network transition function CP-IWF (in the the downstream signal; that is, the signal coming from the line termination unit LT or exchange-side network transition unit CO-IWF to the network termination unit NT1 or the customer-side network transition unit CP-IWF) (F9), loss of signal (LOS) on the line side of the line termination unit LT or the exchange-side network transition unit CO-IWF (F12), operating voltage failure in the network termination unit NT1 or in the customer-side network transition unit CP-IWF (F13), AIS (Alarm Indication Signal) on the line side of the network termination unit NT1 or the customer-side network transition unit CP-IWF, the AIS being generated in the network and forwarded transparently by the line termination unit LT or the exchange-side network transition unit CO-IWF (F13), at the V3 reference point loss of signal (LOS} (F10), at the T reference point loss of signal (LOS) or loss of frame alignment (LFA) (F8), operating voltage failure (if relevant) (F-) Error performance monitoring errored CRC blocks detected on the line side of the network termination unit NT1 or the customer-side network transition unit CP-IWF (F3), errored CRC blocks detected at the T reference point of the network termination unit NT1 or the customer-side network transition unit CP-IWF (F6), CRC error indication received from the customer telephony equipment TE in the E bit (F5), errored CRC blocks detected at the T reference point of the network termination unit NT1 or the customer-side network transition unit CP-IWF and simultaneous reception of a CRC error information from the customer telephony equipment TE (F7).\",\n \"The functions referred to are explained in more detail below with reference to exemplary embodiments.\",\n \"With regard to the above-mentioned functions, the reference specified in parentheses in each case indicates which emulated function F1 to F15 is used for the function concerned.\",\n \"The emulated functions F1 to F15 are explained in more detail below in this order with reference to FIGS.\",\n \"3 to 17.The functions F4, F11 and F14 are not referred to in the above-mentioned list.\",\n \"The error condition on which the function F11 is based is an overlay composed of the error conditions on which functions F8 and F10 are based.\",\n \"The error condition on which function F14 is based is an overlay comprised of the error conditions on which functions F8 and F13 are based.\",\n \"During a normal operating mode of the access digital section and in error conditions which permit the use of time frame signals, i.e.\",\n \"excluding loss of signal (LOS) or loss of frame alignment (LFA) at the T reference point of the network termination unit NT1 or the customer-side network transition unit CP-IWF, the information is transferred transparently in the time slots of the ISDN PRI signal via the access digital section D5.Similarly, the A, Sa4, Sa7 and Sa8 bits are also transferred transparently.\",\n \"The time frame and multi time frame alignment, the CRC-4 bits and the CRC error information (E bit) in both transmission directions as well as the Sa5 and Sa6 bits in the transmission direction toward the exchange ET are generated in the network termination unit NT1 or the customer-side network transition unit CP-IWF.\",\n \"In the case of a loss of signal (LOS) or a loss of time frame alignment (LFA) at the T reference point of the network termination unit NT1 or the customer-side network transition unit CP-IWF, a new time frame is generated.\",\n \"The A bit is set to the value zero.\",\n \"The Sa4, Sa5, Sa7 and Sa8 bits as well as the bits in the time slots TS1 to TS31 are set to the value one.\",\n \"These time frames are also known as substituted frames.\",\n \"A bit sequence consisting of Sa6 bits is used to indicate this error condition.\",\n \"In the exemplary embodiment, instead of a full time-division multiplex frame (TDM Frame) being transferred via the ATM section, each time slot is mapped into a separate AAL2 channel.\",\n \"The time slots TS1 to TS31 contain user information and DSS1 signaling (DSS1—Digital Signaling System Number One).\",\n \"The time slot TS0 transfers error conditions or control information, e.g.\",\n \"loopback requests.\",\n \"As mentioned already, some of the classical PRI-OAM methods (PRI-OAM—Primary Rate Interface-Operation Administration Maintenance) have to be emulated by means of ATM methods, whereby standardized methods defined in ITU-T standard I.610 are used.\",\n \"Thus, for example, the CRC-4 methods, the event indication and/or error indication in the access digital section DS and also the loopback methods are emulated by means of suitable ATM methods.\",\n \"The generation of the time frame and the generation of the multi time frame alignment signal are not possible in the network termination unit NT1 or the customer-side network transition unit CP-IWF in the transmission direction toward the exchange ET.\",\n \"The line termination unit LT or the exchange-side network transition unit CO-IWF handles these functions in the transmission direction toward the exchange ET.\",\n \"In the transmission direction toward the customer telephony equipment TE, the network termination unit NT1 or the customer-side network transition unit CP-IWF generates the time frame, the multi time frame alignment, the CRC-4 bits and the CRC error indication or error information.\",\n \"General Explanations Relating to FIGS.\",\n \"3 to 18 In the upper part of each of the FIGS.\",\n \"3 to 17 there is shown a table in the top row of which the columns of a table header are assigned in each case to the following elements in the order given below: column 1 to the customer telephony equipment TE, which is also referred to as a CPE (Customer Premises Equipment), column 2 to the T reference point, columns 3 and 4 to the customer-side network transition unit CP-IWF, column 5 to the ATM network 10, columns 6 and 7 to the exchange-side network transition unit CO-IWF, column 8 to the reference point V3, and column 10 to the exchange ET, which is also referred to as a service node (Switching Node).\",\n \"Rows 2 and 3 of the table header relate to the assignment of the columns in the transmission direction from the customer telephony equipment TE to the exchange ET. For this transmission direction: column 1 relates to the data transmitted by the customer telephony equipment 12, column 3 relates to the data received in the customer-side network transition unit CP-IWF from the side of the customer telephony equipment TE, column 4 relates to the data transmitted by the customer-side network transition unit CP-IWF into the ATM network 10, column 6 relates to the data received in the exchange-side network transition unit CO-IWF from the ATM network 10, column 7 relates to the data transmitted by the exchange-side network transition unit CO-IWF to the exchange ET, and column 9 relates to the data received in the exchange ET from the exchange-side network transition unit CO-IWF.\",\n \"The positions mentioned are also referred to in this order as the Tup, Tup, ATMup, ATMup, V3up and V3up interface.\",\n \"Rows 2 and 3 of the table header are in each case assigned to the first row of the table body.\",\n \"Rows 4 and 5 of the table header relate to the transfer of data in the transmission direction from the exchange ET to the customer telephony equipment TE.\",\n \"For this transmission direction: the first column of the table relates to the data received in the customer telephony equipment TE and originating from the customer-side network transition unit CP-IWF, column 3 relates to the data transmitted by the customer-side network transition unit CP-IWF to the customer telephony equipment TE, column 4 relates to the data received by the customer-side network transition unit CP-IWF from the ATM network 10, column 6 relates to the data transmitted by the exchange-side network transition unit CO-IWF into the ATM network 10, column 7 relates to the data received in the exchange-side network transition unit CO-IWF from the exchange ET, and column 9 relates to the data transmitted by the exchange ET to the exchange-side network transition unit CO-IWF.\",\n \"The positions mentioned for this transmission direction are also referred to in the given order as the Tdown, Tdown, ATMdown, ATMdown, V3down and V3down interface.\",\n \"The second or last row of the table body is assigned to the transmission direction from the exchange ET to the customer telephony equipment TE.\",\n \"The following symbols are used in the fields of the tables: - no entry because none is necessary to an understanding of the exemplary embodiment or because in accordance with the known standard, a analyze, t transmit unchanged underscore error/event detection point x= .\",\n \".\",\n \".\",\n \"response to an error/event condition ### error/event cause In addition to the entities already explained with reference to FIG.\",\n \"1, FIGS.\",\n \"3 to 17 also show a time-division multiplex data transmission network 12 on both sides of the ATM network 10.Loopbacks Two different loopbacks are supported, a loopback in the exchange-side network transition unit CO-IWF and another loopback in the customer-side network transition unit CP-IWF.\",\n \"The loopbacks are activated as a result of special Sa bit sequences which are explained in more detail below with reference to FIGS.\",\n \"3 and 4.A bit sequence with eight successive code words of the bits Sa6(1), Sa6(2), Sa6(3) and Sa6(4) forms a loopback command which must be detected before further measures are taken.\",\n \"Conversely, the loopback is released if eight successive commands for releasing the loopback or eight other successive signals have been received in which no loopback command has been detected.\",\n \"FIG.\",\n \"3 shows function units and function sequences for activating a loopback in the exchange-side network transition unit CO-IWF.\",\n \"In the case of the loopback in the exchange-side network transition unit CO-IWF, the exchange-side network transition unit CO-IWF transmits the ISDN PRI signal back to the exchange ET.\",\n \"In the transmission direction toward the customer-side network transition unit CP-IWF, the A bit is set to the value one.\",\n \"In the transmission direction toward the exchange ET, the A bit is set to the value one and the Sa5 bit is set to the value zero.\",\n \"The function explained with reference to FIG.\",\n \"3 is also referred to as function F1.FIG.\",\n \"4 shows function units and function sequences for activating a loopback in the customer-side network transition unit CP-IWF.\",\n \"The loopback in the customer-side network transition unit CP-IWF is produced as a result of the insertion of end-to-end F5 loopback cells which are also referred to as F5 LB (Loopback) cells.\",\n \"These cells are inserted by the exchange-side network transition unit CO-IWF after the loopback command relating to the network termination unit NT or the customer-side network transition unit CP-IWF has been detected.\",\n \"At the same time there is set up in the exchange-side network transition unit CO-IWF a local loopback which is similar to the loopback in the case explained with reference to FIG.\",\n \"3 of a loopback in the line termination unit LT or the exchange-side network transition unit CO-IWF.\",\n \"The loopback is regarded as faulty if the LB cells do not return to their point of origin within five seconds.\",\n \"In this case the exchange-side network transition unit CO-IWF manipulates the loopback signal transmitted to the exchange ET. As a result the exchange ET can detect bit errors.\",\n \"The function explained with reference to FIG.\",\n \"4 is also referred to as function F2.Error Performance Monitoring The CRC-4 monitoring is used to provide protection against incorrect time frames (framing) and for error performance monitoring of the access digital section DS.\",\n \"In order to enable service features such as free channel suppression, speech compression, voice activity detection and silence suppression, transparent transmission of the CRC-4 is not possible.\",\n \"For this reason ATM monitoring methods such as VCC (Virtual Channel Connection) performance monitoring and VCC continuity monitoring are used simultaneously for the ATM section of the access digital section DS.\",\n \"The performance monitoring ensures real-time evaluation of the transmission quality for the selected VCCs at segment level or at end-to-end level.\",\n \"In the case of monitoring of the ATM section of the access digital section DS, the end-to-end VCC performance monitoring variant should be used.\",\n \"The performance monitoring is achieved by monitoring blocks from user cells.\",\n \"There are two applications of performance monitoring, namely forward monitoring and backward reporting together or forward monitoring on its own.\",\n \"In order to detect errored blocks of user cells it is sufficient to use only forward monitoring.\",\n \"Performance monitoring is activated during the connection setup as part of the VCC method.\",\n \"In addition the activation or deactivation can be configured for each VCC with the aid of a telecommunication management network (TMN).\",\n \"For communication between the VCC end points, special OAM cells (OAM—Operation Administration Maintenance) are used, namely the FPM cells (FPM—Forward Performance Monitoring) and the BR cells (BR—Backward Reporting).\",\n \"The following parameters are obtained with the aid of performance monitoring: a counter value for the number of transferred user information cells, errored blocks, a counter value for lost user information cells and a counter value for erroneously inserted user information cells within a monitored block consisting of cells.\",\n \"The CRC-4 checksum is computed afresh at the point of transition from the ATM network 10 to the time-division multiplex oriented network 12, i.e.\",\n \"in the transmission direction toward the exchange ET.\",\n \"If errors are detected as a result of VCC performance monitoring and continuity monitoring in the exchange-side network transition unit CO-IWF, this results in a CRC-4 error at the V3up interface of the exchange-side network transition unit CO-IWF (see explanatory remarks relating to FIG.\",\n \"6 below).\",\n \"If a block error is detected by the exchange-side network transition unit CO-IWF, the E bit in the transmission direction toward the exchange ET is set to the value zero.\",\n \"This case is explained in FIG.\",\n \"5.FIG.\",\n \"5 shows the monitoring of the ATM network 10 based on VCC performance monitoring and continuity monitoring at the ATMdown interface of the customer-side network transition unit CP-IWF.\",\n \"Performance errors or continuity errors on the access digital section DS detected on the line side by the customer-side network transition unit CP-IWF form a substitute for CRC-4 monitoring (error report).\",\n \"Error monitoring is simplified.\",\n \"The detection of bit errors in the access digital section DS is possible.\",\n \"There is no possibility of emulating other cases as specified in the ISDN PRI standard, e.g.\",\n \"for the case in which an additional Sa6 bit sequence is to be transmitted if a CRC-4 error threshold of 512 is exceeded.\",\n \"FIG.\",\n \"5 shows the case in which an errored block consisting of user cells has been detected on the basis of the received FPM cells or a loss of continuity (LOC—Loss of Continuity) has been detected as a result of VCC continuity monitoring.\",\n \"FIG.\",\n \"6 shows the monitoring of the ATM network 10 by means of VCC performance monitoring and by means of continuity monitoring at the ATMup interface of the exchange-side network transition unit CO-IWF.\",\n \"The figure shows the sequences when an error is detected in the ATM signal received by the exchange-side network transition unit CO-IWF.\",\n \"VCC continuity monitoring is performed according to ITU-T standard I.610.Of the various functions specified in ITU-T standard I.610, the following are used: continuity monitoring (CC—Continuity Check) is activated during the connection setup, the VC CC cells are transmitted repeatedly at a periodicity of nominally one cell per second irrespective of the number of user cells transferred, a loss of continuity (LOC) is detected if the VCC receiver (sink point) receives no user cell or no continuity monitoring cell wihtin a time interval of 3.5 seconds with a margin of 0.5 seconds.\",\n \"VCC performance monitoring is likewise performed according to ITU-T standard I.610, the following options of ITU-T standard I.610 being used: performance monitoring is activated during the connection establishment, forward performance monitoring is supported, the block size N, after which an FPM cell is inserted, lies outside the regulatory scope of the document, i.e.\",\n \"it is a configurable parameter.\",\n \"The function explained with reference to FIG.\",\n \"5 is also referred to as function F3.The function explained with reference to FIG.\",\n \"6 is also referred to as function F4.In addition, as shown in FIGS.\",\n \"7 and 8, a second CRC-4 section is monitored between the customer-side network transition unit CP-IWF and the customer telephony equipment TE.\",\n \"FIG.\",\n \"7 shows the function sequences when a CRC-4 error is reported by the customer telephony equipment TE.\",\n \"The function explained in FIG.\",\n \"7 is also referred to as function F5.FIG.\",\n \"8 shows function sequences when a CRC-4 error is detected in the Tup signal of the customer-side network transition unit CP-IWF.\",\n \"It is the task of the customer-side network transition unit CP-IWF to detect CRC-4 errors and indicate them to the exchange ET.\",\n \"The associated bit sequences are shown in the table in FIG.\",\n \"8.The function shown with reference to FIG.\",\n \"8 is also referred to as function F6.FIG.\",\n \"9 shows function sequences when a CRC-4 error is reported by the customer telephony equipment TE and a CRC-4 error is simultaneously detected in the Tup signal of the customer-side network transition unit CP-IWF.\",\n \"The function shown with reference to FIG.\",\n \"9 is also referred to as function F7.The error condition explained for function F7 is an overlay of the error conditions which have been explained in connection with functions F5 and F6 with reference to FIGS.\",\n \"7 and 8.Error Indications There follows an overview of the errors that occur and the associated error indications which have to be detected and transmitted in each case by the network transition unit.\",\n \"These error indications are generated in the network termination unit NT1 or the customer-side network transition unit CP-IWF and transferred in the Sa6 bits to the exchange ET. LOS or LFA at the T Reference Point of the NT1/CP-IWF FIG.\",\n \"10 shows function sequences when a loss of signal (LOS) or loss of frame alignment (LFA) is detected at the Tup interface of the customer-side network transition unit CP-IWF.\",\n \"When the loss of signal (LOS) or loss of frame alignment (LFA) is detected, the customer-side network transition unit CP-IWF transmits the Sa6 bit sequence with the value “1100” in the transmission direction toward the exchange-side network transition unit CO-IWF.\",\n \"The exchange-side network transition unit CO-IWF transmits the received Sa6 bit sequence unchanged, i.e.\",\n \"transparently, to the exchange ET. After receiving the Sa6 bit sequence, the exchange ET sets the A bit to the value one and the Sa4 bit to the value zero.\",\n \"These bits are inserted in time slot TS0 in the direction toward the customer.\",\n \"In addition the customer-side network transition unit CP-IWF inserts the E bit containing the value zero into the signal directed toward the customer telephony equipment TE or into the downstream signal.\",\n \"The function explained with reference to FIG.\",\n \"10 is also referred to as function F8.LOS or LFA on the Line Side of the NT1/CP-IWF FIG.\",\n \"11 shows function sequences when an ATM connection error is detected at the ATMdown interface of the customer-side network transition unit CP-IWF.\",\n \"An error in the signal originating from the ATM network 10 is detected in the customer-side network transition unit CP-IWF as a result of end-to-end VCC continuity monitoring, loss of a cell, loss of cell alignment or cell synchronicity or due to a physical connection failure, in particular as a result of a loss of signal (LOS—Loss of Signal) at the XDSL level (XDSL—X-Digital Subscriber Line).\",\n \"The exchange-side network transition unit CO-IWF begins with VCC continuity monitoring (CC—Continuity Check) after the relevant VCC has been established.\",\n \"The VCC continuity monitoring is performed according to ITU standard I.610.Of the various options specified in the I.610 standard, the following are used: continuity monitoring (CC) is activated during the connection establishment, the VC CC cells are transmitted repeatedly at a periodicity of nominally one cell per second irrespective of the number of user cells transferred.\",\n \"The function shown with reference to FIG.\",\n \"11 is also referred to as function F9.LOS at the V3 Reference Point or at the Exchange-Side Network Transition Unit CO-IWF FIG.\",\n \"12 shows function sequences when an ATM connection error occurs at the ATMdown interface, the error having been caused by a loss of signal (LOS) at the exchange-side network transition unit CO-IWF.\",\n \"The function sequences shown in FIG.\",\n \"12 are similar to the function sequences explained with reference to FIG.\",\n \"11.The exchange-side network transition unit CO-IWF deactivates the VCC continuity monitoring (CC) for as long as a loss of signal (LOS) is detected.\",\n \"In addition no user cells, including the cells for transferring the information of time slot TSO, are transmitted.\",\n \"In this way the customer-side network transition unit CP-IWF can detect an ATM connection error.\",\n \"The function explained with reference to FIG.\",\n \"12 is also referred to as function F10.LOS or LFA at the T Reference Point of the NT1/CP-IWF and LOS at the V3 Reference Point of the LT/CO-IWF FIG.\",\n \"13 shows function sequences when a loss of signal (LOS) or loss of frame alignment (LFA) occurs at the T reference point of the customer-side network transition unit CP-IWF and simultaneously a loss of signal (LOS) is detected at the V3 reference point of the exchange-side network transition unit CO-IWF.\",\n \"The function explained with reference to FIG.\",\n \"13 is also referred to as function F11.The error condition explained for function F11 is a combination of the error conditions which have been explained above in connection with the functions F8 and F10 with reference to FIGS.\",\n \"10 and 12 respectively.\",\n \"LOS on the Line Side of the LT/CO-IWF FIG.\",\n \"14 shows function sequences when an ATM connection error occurs in the ATMup signal at the exchange-side network transition unit CO-IWF.\",\n \"If the exchange-side network transition unit CO-IWF detects an error in the ATMup signal as a result of the non-arrival of VCC continuity monitoring cells, loss of cells, loss of cell alignment or physical connection failure, e.g.\",\n \"due to a loss of signal (LOS) at XDSL level, the function sequences shown in FIG.\",\n \"14, which are also referred to as function F12, are executed.\",\n \"AIS at the V3 Reference Point of the CO-IWF and Transfer to the CP-IWF FIG.\",\n \"15 shows function sequences when an AIS (Alarm Indication Signal) is detected at the customer-side network transition unit CP-IWF.\",\n \"The customer-side network transition unit CP-IWF detects the AIS on the line side.\",\n \"The AIS is generated in the time-division multiplex network 12 and forwarded by the exchange-side network transition unit CO-IWF.\",\n \"When AIS alarm data packets are received at the ATMdown interface of the customer-side network transition unit CP-IWF, the customer-side network transition unit CP-IWF transmits an AIS to the customer telephony equipment TE, which is also designated by the acronym CPE (Customer Premises Equipment).\",\n \"The customer telephony equipment TE returns an A bit with the value one, which is transferred unchanged, i.e.\",\n \"transparently, to the exchange ET.\",\n \"The cause for the AIS received in the customer-side network transition unit CP-IWF was an AIS bit sequence inserted by the exchange ET.\",\n \"The function explained with reference to FIG.\",\n \"15 is also referred to as function F13.LOS/LFA at the Tup Interface of the CP-IWF and AIS at the V3 Reference Point of the CO-IWF FIG.\",\n \"16 shows function sequences when an AIS is detected in the ATMdown signal and simultaneously a loss of signal (LOS) or loss of frame alignment (LFA) is detected at the T reference point of the customer-side network transition unit CP-IWF.\",\n \"In this case, the AIS is transferred to the customer-side network transition unit CP-IWF.\",\n \"The function shown with reference to FIG.\",\n \"16 is also referred to as function F14.Function 14 is based on an error case which is a combination of two error cases which have been explained above with reference to functions F8 and F13 and also with reference to FIGS.\",\n \"10 and 15.Operating Voltage Failure in the NT1/CP-IWF FIG.\",\n \"17 shows function sequences when an operating voltage failure is detected at the customer-side network transition unit CP-IWF.\",\n \"When an operating voltage failure is detected by the customer-side network transition unit CP-IWF, the bit values shown in the table in FIG.\",\n \"17 are used.\",\n \"The function shown in FIG.\",\n \"17 is also referred to as function F15.Exemplary Embodiment with Specification of the Basic Text for an ATM Standard There follows an explanation of a further exemplary embodiment in which an expanded basic text is proposed for ATM standard af-vmoa-0145.000.Based on the exemplary embodiments already explained above, it is shown which text sections of the af-vmoa-0145.000 standard are to be replaced or supplemented.\",\n \"New sections are also specified.\",\n \"Changes or addenda are indicated by a note at the beginning of each section, e.g.\",\n \"by “replaces section x.y.z of the af-vmoa-0145.000 standard”.\",\n \"Some sections must be added in the af-vmoa0145.000 standard because the aspects mentioned therein are not covered by the current basic document.\",\n \"This is indicated by a note, e.g.\",\n \"by “new section x.y.z required in af-vmoa-0145.000”.\",\n \"Subsections which contain no such comments are purely informative.\",\n \"The text proposed in the exemplary embodiment still has to be revised for the purpose of drafting a basic text so as to bring it more closely into line with the structure of the af-vmoa-0145.000 standard.\",\n \"Objectives (Replaces Section 1.1 of the af-vmoa-0145.000 Standard) The extension of the subscriber line loop emulation service using ARL2 for ISDN PRI services which are described in this document satisfies a need of the market for an efficient transfer method for carrying ISDN PRI traffic over a broadband subscriber line connection such as e.g.\",\n \"an XDSL line between a customer terminal equipment and the public circuit-switched or line-switched telephone network.\",\n \"The intention is to use the extension for access trunking to the public circuit-switched network with leased lines, as used mainly for the connection of PBXs (PBX—Private Branch Exchange).\",\n \"The classical ISDN PRI is based on conventional time-division multiplex (TDM) methods with extended requirements and OAM principles (OAM—Operation Administration Maintenance) which have been specified for monitoring the circuit-switched connection.\",\n \"For this reason an ISDN PRI emulation service which uses the principles of the ATM/LES must also meet these requirements.\",\n \"The methods required for this are specified in this document.\",\n \"Scope (Replaces Section 1.2 of the af-vmoa-0145.000 Standard) This specification explains the methods which permit the efficient transfer of ISDN PRI (2048 kbps/E1 signal) services over an ATM network between two network transition units (IWF—Interworking Function).\",\n \"The network transition units are located: in the customer area (CP-IWF with the user-side interface at the T reference point, specified in ITU-T standards I.411, I.431 and ETSI ETS 300 011), and in the service provider area (CO-IWF with the local exchange interface at the V3 reference point, specified in ITU-T standard Q.512).\",\n \"Also specified is the use of virtual ATM links via AAL2 for transporting bearer information and signaling.\",\n \"The virtual links used shall be PVCs (Permanent Virtual Circuit).\",\n \"The regulatory scope of this specification covers: the functionality of the network transition units IWF, the relevant aspects of the control level for the subscriber line (loop) emulation service using AAL2 (Loop Emulation Service Using AAL2), the relevant aspects of the user level for the subscriber line (loop) emulation service using AAL2, relevant aspects of the network management level.\",\n \"Reference Model (Replaces Section 1.4 of the af-vmoa-0145.000 Standard) This specification is intended to support the delivery of ISDN PRI services to customer premises via bandwidth-constrained ATM connections such as those provided for example by DSL (Digital Subscriber Line) systems.\",\n \"In this reference model, only those entities are shown that pertain to the ISDN PRI services.\",\n \"In one implementation the equipment providing the CP-IWF function may also include data interfaces toward the customer, e.g.\",\n \"an Ethernet.\",\n \"The data traffic originating from such interfaces or terminating at such interfaces is carried over AAL5 or another appropriate AAL on the same ATM interface to the ATM network as the traffic in the voice band.\",\n \"The service node shown in FIG.\",\n \"18 stands for an exchange of the public circuit-switched telephone network (PSTN—Public Switched Telephone Network) which provides public telephone switching services and leased line services over a narrowband service node interface (SNI).\",\n \"The service node can connect the exchange-side network transition unit CO-IWF via one or more physical interfaces.\",\n \"Alternatively the CO-IWF functionality can be an integral part of the service node with the result that the physical interface between the exchange-side network transition unit CO-IWF and the service node is not visible toward the outside.\",\n \"The physical connection between the CP-IWF and the ATM network is typically provided by a DSL (Digital Subscriber Line), an HFC (Hybrid Fibre Coax), a fiber optic link or a radio link.\",\n \"The ATM network may be a full network, a single ATM switching element or simply a direct interconnection between a CO-IWF and a CP-IWF.\",\n \"The ATM virtual circuits through the ATM network between the CP-IWF and the CO-IWF shall be PVCs or SPVCs (Soft Permanent Virtual Circuits) which carry the following traffic: bearer traffic and signaling using AAL2, where the DSS1 (Digital Signaling System Number One) for controlling the narrowband services is carried in an AAL2 channel within the same ATM VCC as the associated bearer traffic.\",\n \"The CO-IWF and CP-IWF described in this specification are functional units which may be implemented as standalone devices, as parts of larger devices, or distributed among several different devices.\",\n \"This specification does not dictate the implementation of any one of these configurations.\",\n \"CO-IWF Functionality (In Addition to Section 1.7 of the af-vmoa-0145.000 Standard) signaling adaptation (Signaling Interworking) for the transfer of fault messages (failures) and alarm messages (alarms) in order to receive maintenance signaling from the SNI (Service Node Interface) (if present) and from the ATM broadband interfaces and to insert signaling into the SNI and the ATM broadband interfaces.\",\n \"For example, a loopback command originating from the exchange ET is replaced by a VCC loopback and in the other transmission direction errors received in the AAL2 maintenance channel are mapped to error messages (notifications) via Sa bits into the signal which is not used for frame alignment.\",\n \"CP-IWF Functionality (In Addition to Section 1.8 of the af-vmoa-0145.000 Standard) signaling adaptation (Signaling Interworking) for the transfer of failures and alarms in order to receive maintenance signaling from the ISDN PRI interfaces or from the ATM broadband interfaces and to insert signaling into the ISDN PRI interfaces or the ATM broadband interfaces.\",\n \"For example, alarm indications are transferred in the AAL2 maintenance channel.\",\n \"Interfaces Supported (Replaces Section 2 of the af-vmoa-0145.000 Standard) This specification identifies the narrowband interfaces and specifies the ATM interfaces at the CP-IWF and the CO-IWF.\",\n \"The IWFs may also provide other interfaces for management and administration purposes, but these are not specified in this document.\",\n \"IWF Narrowband Interfaces Physical Layer On the physical layer, an IWF (CP- and CO-) should support DS1 or E1 circuits according to the ITU-T standards G.703 and G.704 depending on the application.\",\n \"The channel structure of the primary multiplex interface (Primary Rate Interface Channel Structure) specified in ITU-T standard I.412 should be supported.\",\n \"This means: the primary multiplex B channel structure is supported, i.e.\",\n \"independent B channels operating at 64 kbps.\",\n \"For the E1 interface operating at 2,048 Mbps, the following applies: 30 B+D.\",\n \"For the DS1 interface operating at 1,544 Mbps, the following applies: 23 B+D.\",\n \"The H channel structures are not supported (H0: 384 kbps (kilobits per second), H11: 1536 kbps, H12: 1920 kbps).\",\n \"IWF User-Side and Network-Side Interfaces A CP-IWF must support suitable interfaces for ISDN PRI connection to a telephony equipment in the customer area.\",\n \"The specifications for this interface include: a primary multiplex ISDN interface according to the ITU-T I.431 and ETSI ETS 300 011 standards or equivalent national specifications.\",\n \"A CO-IWF must support appropriate interfaces for ISDN PRI connections to the telephone network (in compliance with ITU-T standard Q.512).\",\n \"Signaling On the signaling layer, an IWF shall support one of the following signaling systems depending on the required application: Signaling systems with common channel: N-ISDN signaling according to ITU-T standards Q.921 and Q.931 (DSS1).\",\n \"N-ISDN according to the ETSI version of DSS1, as specified in the ETSI standards ETS 300 125 and ETS ETSI 300 102-1.Capabilities Supported ATM VCCs for Carrying AAL2 Channels for Supporting a PRI Via an LES (Replaces Section 3.2 of the af-vmoa-0145.000 Standard) An ATM VCC between a CP-IWF and a CO-IWF may include traffic from precisely one ISDN primary multiplex interface.\",\n \"DSS1 signaling for ISDN PRI lines and the B channels of the narrowband interface are carried in the same ATM VCC in different AAL2 channels.\",\n \"Time slot TS0 of an ISDN PRI shall be mapped into the AAL2 channel with the channel identifier CID=16.Time slot TS0 contains the frame alignment signal or the signal not relating to the frame alignment which is used for transferring the maintenance and alarm information.\",\n \"Signaling Between CO-IWF and CP-IWF for the ISDN PRI (Replaces Section 3.3.2 of the af-vmoa-0145.000 Standard) The protocol reference model for a CP-IWF with ISDN PRI in the direction toward the user-side interface is illustrated in FIG.\",\n \"19.The transport of media streams between CP-IWF and CO-IWF is the same as in the case of analog telephony, including the optional use of tone dialing (DTMF Dial Digit Service—Dual Tone Multiple Frequency).\",\n \"The DSS1 signaling is not terminated in the CP-IWF.\",\n \"DSS1 signaling messages are relayed by the CP-IWF from the narrowband D channel across the AAL2 VCC using the transmission/error detection service as specified in ITU-T standard I.366.1.Selection and Changing of Coding (The First Part of this Section is Informative) The same methods are valid as specified in the af-vmoa-0145.000 standard because the support for the H channel structures specified in ITU-T standard I.412 is not provided.\",\n \"These channel structures require time frame integrity.\",\n \"For this reason, AAL2 methods such as silence suppression and speech compression can only be used simultaneously for the entire H channel.\",\n \"Thus, for example, a change of coding profile within a profile would only be possible if it simultaneously affects all bearer channels.\",\n \"This means that in the case of an H0, H11 or H12 channel the profile change must take place for all 6, 23 or 30 B channels simultaneously.\",\n \"(The Following Text Supplements Section 3.4.2 of the af-vmoa-0145.000 Standard) Time slot TS0 of the ISDN PRI is mapped into the AAL2 channel with the CID=16.The SSCS (Service Specific Convergence Sublayer) specified in ITU-T standard I.366.2 is used in this channel with PCM64 coding and a change of coding profile is not permitted because the information contained in time slot TS0 must be transferred transparently.\",\n \"Consequently no speech compression and no silence suppression should be used for this channel.\",\n \"Application Identifier (AppId) (Supplements Section 4.1.1 of the af-vmoa-0145.000 Standard) In addition an application identifier must be specified for: the subscriber line (loop) emulation service using DSS1 to support PRI with ELCP (Emulated Loop Control Protocol).\",\n \"SSCS Type The SSCS type to be used on each channel of an AAL2 VCC will vary according to the usage of that channel.\",\n \"Channels that are used to carry media streams (ISDN B channels) shall use the SCCS specified in ITU-T standard I.366.2.The channels that are used to carry the control and management level traffic (ELCP, DSS1 in the ISDN D channels and LES-EOC) shall use the SCCS specified in ITU-T standard I.366.1.The SCCS specified in ITU-T standard ITU-T I.366.2 shall be used to carry additional failure and alarm messages such as are included in time slot TS0 of the ISDN PRI application, but without speech compression and/or silence suppression and VAD methods (Voice Activity Detection).\",\n \"CID Allocation (Supplements Section 4.4.1 of the af-vmoa-0145.000 Standard) In the case of support for ISDN PRI the CID with the value sixteen shall be used for transferring time slot TS0 of the ISDN PRI signal.\",\n \"This CID is assigned during system startup, with the channel being activated at the same time.\",\n \"Signaling and Control Methods The information contained in this section must be assigned to the corresponding sections of the basic text in order to fit into the structure of the af-vmoa-0145.000 standard.\",\n \"Additional signaling and control methods for ISDN PRI are disclosed.\",\n \"Signaling Methods for the ISDN PRI The ISDN PRI consists of: 30 time slots, numbered from 1 to 15 and 17 to 31, for transferring the B channels, a signaling time slot with the number 16 for transporting the DSS1 signaling, and a time slot with the number 0 for transferring the time frame alignment of the signal for the CRC multi time frame alignment, the CRC, and maintenance information in the case of an E1 interface.\",\n \"Each active B channel, the signaling channel (DSS1 signaling), the ELCP protocol and the channel containing the frame alignment signal or the signal not relating to the frame alignment shall be transferred in a separate AAL2 channel of the same VCC.\",\n \"B channels are allocated dynamically to the AAL2 channels using the ELCP according to the call status.\",\n \"The AAL2 channels for the ELCP and the frame alignment signal and for the signal not relating to the frame alignment are allocated statically.\",\n \"For DSS1 signaling, the AAL2 channel is allocated dynamically by the CO-IWF using an ALLOCATION message, as in the case of the ISDN BRI.\",\n \"For the signal relating to the frame alignment and for the signal not relating to the frame alignment (TS0), the AAL2 channel is identified by the CID with the value sixteen.\",\n \"Detection of Unused Channels (Idle Channel Detection) The CO-IWF receives no explicit indication of the seizure or release of a channel, as in the case of the LES and a V5 or a GR303 SNI.\",\n \"The CO-IWF must therefore be informed by means of other measures.\",\n \"The proposed solution is that the CO-IWF detects the call status “Idle” as a result of the monitoring of the DSS1 messages in time slot sixteen of the narrowband interface.\",\n \"Based on the messages for call setup and release, the CO-IWF will seize or release the necessary ARL2 channels using ELCP.\",\n \"The signaling information itself is transferred transparently between the IWFs using the SSSAR (Service Specific Segmentation And Reassembly) and the SSTED methods (Service Specific Transmission Error Detection) in accordance with the af-vmoa-0145.000 standard.\",\n \"FIG.\",\n \"20 shows the protocol reference model for the CO-IWF.\",\n \"ISDN PRI Specific OAM Methods and Tables of the Alarm States (A New Section is Required in the af-vmoa-0145.000 Standard) Loopbacks (The text contained in the Loopbacks section above should be inserted here, i.e.\",\n \"including the explanations concerning the functions F1 and F2 as well as FIGS.\",\n \"3 and 4).\",\n \"Error Monitoring (The text contained in the Error monitoring section above should be inserted here, i.e.\",\n \"including the explanations concerning the functions F3 to F7 as well as FIGS.\",\n \"5 to 9).\",\n \"Error Indication (The text contained in the Error indications section above should be inserted here, i.e.\",\n \"including the explanations concerning the functions F8 to F15 and FIGS.\",\n \"10 to 17).\",\n \"List of Reference Characters DS Access digital section 4, 6 CRC-4 methods TE, CPE Customer telephony equipment, Customer Premises Equipment NT1 Network termination unit LT Line termination unit ET, SN Exchange, Service Node CRC-4 Error code A A bit E E bit Sa4 to Sa8 Sa4 to Sa8 bit CP-IWF Customer-side network transition unit (Customer Premises Interworking Function CO-IWF Exchange-side network transition unit (Central Office Interworking Function 10 ATM network 12 Time-division multiplex network LOS Loss of Signal LFA Loss of Frame Alignment) LOC Loss of Continuity CC Continuity Check) AIS Alarm Indication Signal TS0 Clock start time slot a analyze t forward transparently FPM Forward Performance Monitoring ATM Asynchronous Transfer Mode F1 to F15 Function V3, T Reference point I, II Sub-multi time frame LB Loopback DS1 Connection type (1,544 Mbit/s) E1 Connection type (2,048 Mbit/s) AIS Alarm Indication Signal AUXP Auxiliary Pattern\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473969"}}},{"rowIdx":773,"cells":{"text":{"kind":"list like","value":[["Switching contact arrangement for a power switch","The invention relates to a switching contact arrangement for a power switch, which is characterized in that a plurality of contact levers are pivoted on a contact support (30) by means of a bearing pin (37) and that the contact support (30) is provided with at least three support elements (32a, 32b, 31) for radially supporting the bearing pin.","In order to simplify production of the contact support, at least three of the support elements (32a, 32b, 31) are configured as a one-piece shaped element that forms the contact support (30).","At least one of the support elements can have a metal element (31) that is at least partially embedded in a plastic shaped element (43) of the contact support (30).","Said metal element (31) can be configured as a sheet metal part."],["1.A switching contact arrangement for an electrical switch, comprising: a contact mount; at least two contact levers, pivotably attached to the mount via a bearing bolt, wherein the contact mount includes at least three supporting elements adapted to radially support the bearing bolt, and wherein at least three of the supporting elements are produced integrally in the form of at least part of a molding forming the contact mount.","2.The switching contact arrangement as claimed in claim 1, wherein the molding is at least partially in the form of a plastic molding.","3.The switching contact arrangement as claimed in claim 2, wherein at least one metallic reinforcement element is at least partially embedded in the plastic molding.","4.The switching contact arrangement as claimed in claim 2, wherein at least one of the supporting elements has a metal part which is at least partially embedded in the plastic molding.","5.The switching contact arrangement as claimed in claim 4, wherein the metal part is in the form of a metal sheet, a first subregion of which, including undercuts, is embedded in the plastic molding, and a second subregion of which, provided with a hole for the bearing bolt, projects out of the plastic molding.","6.The switching contact arrangement as claimed in claim 4, wherein, when the contact mount is coupled to a switching shaft via a metallic coupling element, the supporting elements which contain a metal part are at a distance from the coupling element in the axial direction of the bearing bolt.","7.The switching contact arrangement as claimed in claim 2, wherein when the contact mount is equipped with a shielding body on at least one of the supporting elements, a holder is provided for the shielding body.","8.The switching contact arrangement as claimed in claim 3, wherein at least one of the supporting elements has a metal part which is at least partially embedded in the plastic molding.","9.The switching contact arrangement as claimed in claim 8, wherein the metal part is in the form of a metal sheet, a first subregion of which, including undercuts, is embedded in the plastic molding, and a second subregion of which, provided with a hole for the bearing bolt, projects out of the plastic molding.","10.The switching contact arrangement as claimed in claim 5, wherein, when the contact mount is coupled to a switching shaft via a metallic coupling element, the supporting elements which contain a metal part are at a distance from the coupling element in the axial direction of the bearing bolt.","11.The switching contact arrangement as claimed in claim 8, wherein, when the contact mount is coupled to a switching shaft via a metallic coupling element, the supporting elements which contain a metal part are at a distance from the coupling element in the axial direction of the bearing bolt.","12.The switching contact arrangement as claimed in claim 9, wherein, when the contact mount is coupled to a switching shaft via a metallic coupling element, the supporting elements which contain a metal part are at a distance from the coupling element in the axial direction of the bearing bolt.","13.The switching contact arrangement as claimed in claim 3, wherein when there is a contact mount equipped with a shielding body on at least one of the supporting elements, a holder is provided for the shielding body.","14.The switching contact arrangement as claimed in claim 4, wherein when there is a contact mount equipped with a shielding body on at least one of the supporting elements, a holder is provided for the shielding body.","15.The switching contact arrangement as claimed in claim 5, wherein when there is a contact mount equipped with a shielding body on at least one of the supporting elements, a holder is provided for the shielding body.","16.The switching contact arrangement as claimed in claim 6, wherein when there is a contact mount equipped with a shielding body on at least one of the supporting elements, a holder is provided for the shielding body.","17.A switching contact arrangement for a switch, comprising: at least two contact levers, pivotably attached to a contact support via a bearing pin, wherein the contact support includes at least three elements adapted to radially support the bearing pin, and wherein at least three of the elements are configured as a one piece shaped element forming the contact support.","18.The switching contact arrangement as claimed in claim 17, wherein the one piece element is at least partially formed as a plastic molding.","19.The switching contact arrangement as claimed in claim 18, wherein at least one metallic reinforcement element is at least partially embedded in the plastic molding.","20.The switching contact arrangement as claimed in claim 18, wherein at least one of the supporting elements includes a metal part which is at least partially embedded in the plastic molding.","21.The switching contact arrangement as claimed in claim 20, wherein the metal part is in the form of a metal sheet, a first subregion of which, including undercuts, is embedded in the plastic molding, and a second subregion of which, provided with a hole for the bearing pin, projects out of the plastic molding.","22.The switching contact arrangement as claimed in claim 20, wherein, when the contact mount is coupled to a switching shaft via a metallic coupling element, the supporting elements which contain a metal part are at a distance from the coupling element in the axial direction of the bearing pin.","23.The switching contact arrangement as claimed in claim 18, wherein when the contact mount is equipped with a shielding body on at least one of the supporting elements, a holder is provided for the shielding body."],["<SOH> BACKGROUND OF THE INVENTION <EOH>In a known electrical switch of this type, in which two or more contact levers which form the contact are held on the contact mount by way of a bearing bolt such that they can pivot, the contact mount has at least three supporting elements in order to support the bearing bolt radially (E 0 222 686 B1).","The contact mount in this case includes a metal frame, which is formed from two side walls and from two or more bolts which connect the side walls.","The metal frame is hinged via a coupling bolt on an insulating coupling element, which is used for coupling the contact mount to a switch drive.","Two of the supporting elements, which are associated with the ends of the bearing bolt, are in this case formed by the side walls of the metal frame.","In order to prevent undesirable radial bending of the bearing bolt for the contact levers with as little complexity in terms of additional material as possible, two intermediate bearings for this contact mount, which are arranged between adjacent contact levers in the axially central region of the bearing bolt, form additional supporting elements by being hinged on the coupling bolt.","In this case, aperture openings are required for the contact mount, for the intermediate bearings to pass through to the coupling bolt.","Supporting elements which are integrated in this way in addition to the two outer supporting elements in the contact mount must be positioned for installation of the bearing bolt, owing to their capability to pivot about the coupling bolt."],["<SOH> SUMMARY OF THE INVENTION <EOH>Against the background of an electrical switch, an embodiment of the invention is based on an object of simplifying the production and installation of the contact mount.","According to an embodiment of the invention, an object may be achieved in that at least three of the supporting elements are in the form of part of a molding which forms the contact mount and is produced integrally.","For the purposes of an embodiment of the invention, the expression an integrally produced molding should be understood as being a part in which two or more functional elements are connected in the course of a molding process, such as a stamping, injection-compression molding, casting, injection molding, compression molding or sintering process, to form a single component which is assembled such that it cannot be disconnected for installation purposes.","In the case of a refinement such as this, the three supporting elements are integrated rigidly in a predetermined position in the contact mount, as part of it.","In this case, the three supporting elements are actually aligned with the axis of the bearing bolt during the production of the contact mount so that no tilting of the bearing bolt caused by tolerance discrepancies will in practice occur during operation of the switch.","A bearing bolt which is supported in this way is also subject to only a small maximum amount of bending when high short-circuit or surge currents occur, and thus has a good capability to withstand high short-circuit and surge currents.","The novel switching contact arrangement may have a large number of contact levers, which are each subject to an individual tolerance discrepancy from a given nominal size, and intermediate bearings, which are possibly likewise subject to an individual tolerance discrepancy from their nominal size, but which may be part of the contact levers, since the number of contact levers is subdivided into subsets.","Each of these is arranged axially bounded between two adjacent supporting elements.","This axial bounding of the subsets of contact levers limits any axial movement of the contact levers in one subset, owing to the current forces which act between them, to the axial section of the bearing bolt which is bounded by the respective supporting elements.","The maximum amount of movement is not greater than the sum of all the individual tolerance discrepancies of the contact levers and of the intermediate bearings, which may be present, in this subset.","This makes it possible to geometrically associate the contact levers with contact force springs such that their spring force is not reduced by bending or tilting.","The geometrically accurate association between the contact force springs and the contact levers thus also contributes to increasing the capability of the switching contact arrangement to withstand short-circuit and surge currents.","If the molding is at least partially in the form of a plastic molding, then there is no need for the coupling bolt to have an electrically insulating configuration.","The mechanical strength of a plastic molding such as this can be increased by embedding at least one reinforcement element at least partially in the plastic molding.","A thermosetting plastic which, for example, has fiber reinforcement is typically used for the plastic molding and a nonmagnetic steel, for example, is used for the reinforcement element.","As an alternative to this, other pure plastics or, for example, plastics reinforced by ceramic or glass fibers can also be used for the plastic molding, and other metals or metal sheets can be used for the reinforcement element.","One preferred refinement of the novel switching contact arrangement provides for at least one of the supporting elements to have a metal part which is at least partially embedded in the plastic molding.","A metal part such as this may be part of the reinforcement element, thus at the same time increasing the mechanical strength of that part of the contact mount which forms the supporting element.","If the metal part is in the form of a metal sheet, for example composed of nonmagnetic sheet steel, a first subregion of which, which has undercuts, is embedded in the plastic molding and a second subregion of which, which is provided with a hole for the bearing bolt, projects out of the plastic molding.","This then advantageously allows the cross section of the supporting element to be reduced such that it is no broader than the distance between the contact levers that is required for separation of the contact levers and thus does not lead to any additional broadening of the contact mount.","A further advantageous refinement of the novel switching contact arrangement provides for supporting elements which contain the metal parts to be at a distance from the coupling element in the axial direction of the bearing bolt if the contact mount is coupled to a switching shaft, which can be rotated using a switch drive, via a metallic coupling element.","This makes it possible to avoid accidental energizing and problems relating to the insulation between the contact mount and the switch drive, in a simple manner.","In this refinement, the entire available material depth of the contact mount transversely with respect to the bearing bolt can be used for the rigid embedding of a first subregion of a supporting element which is in the form of a metal sheet.","If a holder for the shielding body is provided on at least one of the supporting elements for a contact mount which is equipped with the shielding body, then this provides additional support for the shielding body against the gas pressure which occurred during switching.","In a refinement such as this, side mounting limbs, which rest on the contact mount, are designed to be smaller owing to the reduced load, or may possibly be omitted."],["This application is the national phase under 35 U.S.C.","§ 371 of PCT International Application No.","PCT/DE02/01250 which has an International filing date of Mar.","28, 2002, which designated the United States of America and which claims priority on German Patent Application numbers DE 101 17 844.1 filed Apr.","4, 2001 and DE 201 18 493.1 filed Nov. 7, 2001, the entire contents of which are hereby incorporated herein by reference.","FIELD OF THE INVENTION The invention generally relates to the field of electrical/power switches, and is preferably applicable to the design configuration of a rigid member which is used as a contact mount for a contact.","BACKGROUND OF THE INVENTION In a known electrical switch of this type, in which two or more contact levers which form the contact are held on the contact mount by way of a bearing bolt such that they can pivot, the contact mount has at least three supporting elements in order to support the bearing bolt radially (E 0 222 686 B1).","The contact mount in this case includes a metal frame, which is formed from two side walls and from two or more bolts which connect the side walls.","The metal frame is hinged via a coupling bolt on an insulating coupling element, which is used for coupling the contact mount to a switch drive.","Two of the supporting elements, which are associated with the ends of the bearing bolt, are in this case formed by the side walls of the metal frame.","In order to prevent undesirable radial bending of the bearing bolt for the contact levers with as little complexity in terms of additional material as possible, two intermediate bearings for this contact mount, which are arranged between adjacent contact levers in the axially central region of the bearing bolt, form additional supporting elements by being hinged on the coupling bolt.","In this case, aperture openings are required for the contact mount, for the intermediate bearings to pass through to the coupling bolt.","Supporting elements which are integrated in this way in addition to the two outer supporting elements in the contact mount must be positioned for installation of the bearing bolt, owing to their capability to pivot about the coupling bolt.","SUMMARY OF THE INVENTION Against the background of an electrical switch, an embodiment of the invention is based on an object of simplifying the production and installation of the contact mount.","According to an embodiment of the invention, an object may be achieved in that at least three of the supporting elements are in the form of part of a molding which forms the contact mount and is produced integrally.","For the purposes of an embodiment of the invention, the expression an integrally produced molding should be understood as being a part in which two or more functional elements are connected in the course of a molding process, such as a stamping, injection-compression molding, casting, injection molding, compression molding or sintering process, to form a single component which is assembled such that it cannot be disconnected for installation purposes.","In the case of a refinement such as this, the three supporting elements are integrated rigidly in a predetermined position in the contact mount, as part of it.","In this case, the three supporting elements are actually aligned with the axis of the bearing bolt during the production of the contact mount so that no tilting of the bearing bolt caused by tolerance discrepancies will in practice occur during operation of the switch.","A bearing bolt which is supported in this way is also subject to only a small maximum amount of bending when high short-circuit or surge currents occur, and thus has a good capability to withstand high short-circuit and surge currents.","The novel switching contact arrangement may have a large number of contact levers, which are each subject to an individual tolerance discrepancy from a given nominal size, and intermediate bearings, which are possibly likewise subject to an individual tolerance discrepancy from their nominal size, but which may be part of the contact levers, since the number of contact levers is subdivided into subsets.","Each of these is arranged axially bounded between two adjacent supporting elements.","This axial bounding of the subsets of contact levers limits any axial movement of the contact levers in one subset, owing to the current forces which act between them, to the axial section of the bearing bolt which is bounded by the respective supporting elements.","The maximum amount of movement is not greater than the sum of all the individual tolerance discrepancies of the contact levers and of the intermediate bearings, which may be present, in this subset.","This makes it possible to geometrically associate the contact levers with contact force springs such that their spring force is not reduced by bending or tilting.","The geometrically accurate association between the contact force springs and the contact levers thus also contributes to increasing the capability of the switching contact arrangement to withstand short-circuit and surge currents.","If the molding is at least partially in the form of a plastic molding, then there is no need for the coupling bolt to have an electrically insulating configuration.","The mechanical strength of a plastic molding such as this can be increased by embedding at least one reinforcement element at least partially in the plastic molding.","A thermosetting plastic which, for example, has fiber reinforcement is typically used for the plastic molding and a nonmagnetic steel, for example, is used for the reinforcement element.","As an alternative to this, other pure plastics or, for example, plastics reinforced by ceramic or glass fibers can also be used for the plastic molding, and other metals or metal sheets can be used for the reinforcement element.","One preferred refinement of the novel switching contact arrangement provides for at least one of the supporting elements to have a metal part which is at least partially embedded in the plastic molding.","A metal part such as this may be part of the reinforcement element, thus at the same time increasing the mechanical strength of that part of the contact mount which forms the supporting element.","If the metal part is in the form of a metal sheet, for example composed of nonmagnetic sheet steel, a first subregion of which, which has undercuts, is embedded in the plastic molding and a second subregion of which, which is provided with a hole for the bearing bolt, projects out of the plastic molding.","This then advantageously allows the cross section of the supporting element to be reduced such that it is no broader than the distance between the contact levers that is required for separation of the contact levers and thus does not lead to any additional broadening of the contact mount.","A further advantageous refinement of the novel switching contact arrangement provides for supporting elements which contain the metal parts to be at a distance from the coupling element in the axial direction of the bearing bolt if the contact mount is coupled to a switching shaft, which can be rotated using a switch drive, via a metallic coupling element.","This makes it possible to avoid accidental energizing and problems relating to the insulation between the contact mount and the switch drive, in a simple manner.","In this refinement, the entire available material depth of the contact mount transversely with respect to the bearing bolt can be used for the rigid embedding of a first subregion of a supporting element which is in the form of a metal sheet.","If a holder for the shielding body is provided on at least one of the supporting elements for a contact mount which is equipped with the shielding body, then this provides additional support for the shielding body against the gas pressure which occurred during switching.","In a refinement such as this, side mounting limbs, which rest on the contact mount, are designed to be smaller owing to the reduced load, or may possibly be omitted.","BRIEF DESCRIPTION OF THE DRAWINGS The present invention will become more fully understood from the detailed description of preferred embodiments given hereinbelow and the accompanying drawings, which are given by way of illustration only and thus are not limitative of the present invention, wherein FIGS.","1 to 9 show a number of exemplary embodiments of the novel switching contact arrangement and wherein: FIG.","1 shows a schematic section illustration of a low-voltage circuit breaker with a switching contact arrangement which comprises a stationary contact assembly and a moving contact assembly, FIG.","2 shows a moving contact assembly with a first embodiment of a contact mount which is at least partially in the form of a plastic molding, FIG.","3 shows a reinforcement element, which may be embedded in the plastic molding of the contact mount shown in FIG.","2, FIG.","4 shows a second embodiment of a contact mount which is at least partially in the form of a plastic molding, and in which supporting elements in the form of a metal sheet are partially embedded in the plastic molding, FIG.","5 shows a section illustration, transversely with respect to the direction of the bearing bolt, through the contact mount shown in FIG.","4, FIG.","6 shows a reinforcement element which may be embedded in the plastic molding of the contact mount in FIG.","4, FIG.","7 shows a section illustration, transversely with respect to the direction of the bearing bolt, through a contact mount with an embedded reinforcement element as shown in FIG.","6, FIG.","8 shows a further section illustration through the contact mount illustrated in FIG.","4 along the line A-A in FIG.","5, and FIG.","9 shows the contact lever mount as shown in FIG.","4 with a shielding body.","DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The electrical switch which is shown in FIG.","1 and is used in low-voltage power supply systems for voltage ranges up to about 1 000 V has a switch pole enclosure 1 in which switching chambers 2 are formed alongside one another, depending on the number of poles required.","A switching shaft 4, which can be rotated by way of a switch drive 3, is used to operate the switching contact arrangement jointly, each of which includes a stationary contact assembly 5 and a moving contact assembly 6.For this purpose, two levers 7 which project radially from the switching shaft 4 (see FIG.","8) are coupled to a metallic coupling element which is hinged on the moving contact assembly 6.The contact assemblies 5, 6 are connected in a known manner to externally accessible connecting rails 9, 10.Two exemplary embodiments 6a and 6b of the moving contact assembly 6 will be explained in the following text with reference to FIGS.","2 and 3 and, respectively, FIGS.","4 to 9.As can be seen in more detail from FIG.","2, the moving contact assembly 6a has a contact mount 11 which has a plastic molding 12 in the form of an integrally produced molding, which was formed in the course of a stamping process, with a reinforcement element as shown in FIG.","3 being embedded in it.","The contact mount is mounted in the enclosure 1 (see FIG.","1) such that it can pivot, and can be moved via the switching shaft 4 and by use of the switch drive 3, of which FIG.","1 shows only one drive run 15 that is supported on a spring stalk 14 relative to the stationary contact assembly 5 to a connected position and to a disconnected position.","Two or more contact levers 16, 17, which are arranged parallel to one another, on the contact mount 11 can pivot relative to the contact mount 11 about a bearing bolt 18.Contact force springs 19 (see FIG.","1) ensure that the contact levers 16, 17 are prestressed in the direction of the stationary contact assembly 5.Flexible conductors 20 in the form of braids or strips are used for connecting the contact levers 16, 17 to the lower connecting rail 10 in such a way as to guarantee that the contact levers 16, 17 and the contact mount 11 can move without any impediment during the switching movements.","The number of contact levers 16, 17 which are fitted to the contact mount 11 depends on the magnitude of the current which the circuit breaker is intended to carry during operation.","As can be seen from FIG.","2, of the total of 22 contact levers that are provided, 8 contact levers 16 are designed to be shorter and have only one contact area 21, which have no leading contact area 22 and no arcing horn 23 in the same way as the other contact levers 17.During operation, all the contact levers are held between side pieces 24a, 24b of the contact mount 11, which point transversely with respect to the bearing bolt 18.These side pieces 24a, 24b, which are provided with holding openings 25a, 25b for the bearing bolt 18 form a first and a second supporting element for the ends of the bearing bolt, via which the bearing bolt is positioned axially and is supported radially.","A part 29a or 29b of the reinforcement element 13 (see FIG.","3) can extend in each of these side pieces, and has an aperture 27a or 27b, respectively, for the bearing bolt.","In the downward direction, the side pieces 24a, 24b merge into bearing arms 26 for the contact mount 11.The relatively large width of the switching contact arrangement indicates that the section of the bearing bolt which runs between the two side supporting elements 24a, 24b is subjected to a relatively severe bending load when further forces in addition to the forces of the contact force springs 19 are caused by a heavy current, such as a short-circuit or surge current, when the switching contact arrangement is in the closed state.","Bending of this section of the bearing bolt is prevented by way of an additional, third supporting element, which supports the bearing bolt axially in the center.","This third supporting element is formed by a contact mount rib 28, which is provided with a holding opening 25c (which cannot be seen in the figure) for the bearing bolt 18 and points transversely with respect to the bearing bolt, with a metal part 29c (which is completely embedded in the plastic molding 12 and has an aperture 27c for the bearing bolt) extending in the rib 28 and being part of the reinforcement element 13 (see FIG.","3).","Of the second exemplary embodiment 6b of the moving contact group, FIG.","4 shows only a second embodiment 30 of the contact mount.","In this contact mount 30, two supporting elements, which are in the form of metal sheets 31, are used to radially support that section of the bearing bolt which runs between two supporting elements that are in the form of side pieces 32a, 32b.","A first subregion 33 of the two central supporting elements 31 is embedded in the plastic molding 34 of the contact mount 30, and together with a second subregion 35 of the two central supporting elements 31, projects out of the plastic molding.","As can be seen in FIG.","5, the metal sheets 31 have undercuts 36 in the first subregion 33 which is embedded in the plastic molding, and these are used to anchor the respective metal sheet in the plastic molding securely even when the bearing bolt is subjected to a high bending load.","The second subregion 35, which is provided with a hole 38 for the bearing bolt 37 to pass through, also has a recess 39, which is used to hold a shielding body 40 that is not shown in any more detail in FIG.","9.As is shown in FIG.","7, two reinforcement elements 41, 42 may be embedded in the plastic molding of the contact mount, and one of these is illustrated in FIG.","6.As can be seen from FIG.","8, the two metal sheets 31 are at a distance from the metallic coupling element 8 (see FIG.","1) in the axial direction, in order to avoid accidental energizing between the bearing bolt (which is at a low-voltage potential) of the contact levers and the metallic coupling element 8 (which is at ground potential), and thus the switch drive.","As can be seen from FIG.","9, tongues 43 are integrally formed on the shielding body 40, which protects the pivoting area of the contact mount 30 and further switch parts (which are not shown in any more detail but are arranged underneath the contact areas 21, 22 (see FIG.","1)) against erosion products that fall out and against condenser switching gases, and these tongues 43 engage in the recesses 39 which are provided on the metal sheets (see also FIG.","5).","In consequence, the shielding body 40 is supported against the gas pressure which occurs during switching processes, in such a way that its mounting limbs 44a, 44b, which are held in the side on the contact mount, are less severely loaded.","Exemplary embodiments being thus described, it will be obvious that the same may be varied in many ways.","Such variations are not to be regarded as a departure from the spirit and scope of the present invention, and all such modifications as would be obvious to one skilled in the art are intended to be included within the scope of the following claims."]],"string":"[\n [\n \"Switching contact arrangement for a power switch\",\n \"The invention relates to a switching contact arrangement for a power switch, which is characterized in that a plurality of contact levers are pivoted on a contact support (30) by means of a bearing pin (37) and that the contact support (30) is provided with at least three support elements (32a, 32b, 31) for radially supporting the bearing pin.\",\n \"In order to simplify production of the contact support, at least three of the support elements (32a, 32b, 31) are configured as a one-piece shaped element that forms the contact support (30).\",\n \"At least one of the support elements can have a metal element (31) that is at least partially embedded in a plastic shaped element (43) of the contact support (30).\",\n \"Said metal element (31) can be configured as a sheet metal part.\"\n ],\n [\n \"1.A switching contact arrangement for an electrical switch, comprising: a contact mount; at least two contact levers, pivotably attached to the mount via a bearing bolt, wherein the contact mount includes at least three supporting elements adapted to radially support the bearing bolt, and wherein at least three of the supporting elements are produced integrally in the form of at least part of a molding forming the contact mount.\",\n \"2.The switching contact arrangement as claimed in claim 1, wherein the molding is at least partially in the form of a plastic molding.\",\n \"3.The switching contact arrangement as claimed in claim 2, wherein at least one metallic reinforcement element is at least partially embedded in the plastic molding.\",\n \"4.The switching contact arrangement as claimed in claim 2, wherein at least one of the supporting elements has a metal part which is at least partially embedded in the plastic molding.\",\n \"5.The switching contact arrangement as claimed in claim 4, wherein the metal part is in the form of a metal sheet, a first subregion of which, including undercuts, is embedded in the plastic molding, and a second subregion of which, provided with a hole for the bearing bolt, projects out of the plastic molding.\",\n \"6.The switching contact arrangement as claimed in claim 4, wherein, when the contact mount is coupled to a switching shaft via a metallic coupling element, the supporting elements which contain a metal part are at a distance from the coupling element in the axial direction of the bearing bolt.\",\n \"7.The switching contact arrangement as claimed in claim 2, wherein when the contact mount is equipped with a shielding body on at least one of the supporting elements, a holder is provided for the shielding body.\",\n \"8.The switching contact arrangement as claimed in claim 3, wherein at least one of the supporting elements has a metal part which is at least partially embedded in the plastic molding.\",\n \"9.The switching contact arrangement as claimed in claim 8, wherein the metal part is in the form of a metal sheet, a first subregion of which, including undercuts, is embedded in the plastic molding, and a second subregion of which, provided with a hole for the bearing bolt, projects out of the plastic molding.\",\n \"10.The switching contact arrangement as claimed in claim 5, wherein, when the contact mount is coupled to a switching shaft via a metallic coupling element, the supporting elements which contain a metal part are at a distance from the coupling element in the axial direction of the bearing bolt.\",\n \"11.The switching contact arrangement as claimed in claim 8, wherein, when the contact mount is coupled to a switching shaft via a metallic coupling element, the supporting elements which contain a metal part are at a distance from the coupling element in the axial direction of the bearing bolt.\",\n \"12.The switching contact arrangement as claimed in claim 9, wherein, when the contact mount is coupled to a switching shaft via a metallic coupling element, the supporting elements which contain a metal part are at a distance from the coupling element in the axial direction of the bearing bolt.\",\n \"13.The switching contact arrangement as claimed in claim 3, wherein when there is a contact mount equipped with a shielding body on at least one of the supporting elements, a holder is provided for the shielding body.\",\n \"14.The switching contact arrangement as claimed in claim 4, wherein when there is a contact mount equipped with a shielding body on at least one of the supporting elements, a holder is provided for the shielding body.\",\n \"15.The switching contact arrangement as claimed in claim 5, wherein when there is a contact mount equipped with a shielding body on at least one of the supporting elements, a holder is provided for the shielding body.\",\n \"16.The switching contact arrangement as claimed in claim 6, wherein when there is a contact mount equipped with a shielding body on at least one of the supporting elements, a holder is provided for the shielding body.\",\n \"17.A switching contact arrangement for a switch, comprising: at least two contact levers, pivotably attached to a contact support via a bearing pin, wherein the contact support includes at least three elements adapted to radially support the bearing pin, and wherein at least three of the elements are configured as a one piece shaped element forming the contact support.\",\n \"18.The switching contact arrangement as claimed in claim 17, wherein the one piece element is at least partially formed as a plastic molding.\",\n \"19.The switching contact arrangement as claimed in claim 18, wherein at least one metallic reinforcement element is at least partially embedded in the plastic molding.\",\n \"20.The switching contact arrangement as claimed in claim 18, wherein at least one of the supporting elements includes a metal part which is at least partially embedded in the plastic molding.\",\n \"21.The switching contact arrangement as claimed in claim 20, wherein the metal part is in the form of a metal sheet, a first subregion of which, including undercuts, is embedded in the plastic molding, and a second subregion of which, provided with a hole for the bearing pin, projects out of the plastic molding.\",\n \"22.The switching contact arrangement as claimed in claim 20, wherein, when the contact mount is coupled to a switching shaft via a metallic coupling element, the supporting elements which contain a metal part are at a distance from the coupling element in the axial direction of the bearing pin.\",\n \"23.The switching contact arrangement as claimed in claim 18, wherein when the contact mount is equipped with a shielding body on at least one of the supporting elements, a holder is provided for the shielding body.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>In a known electrical switch of this type, in which two or more contact levers which form the contact are held on the contact mount by way of a bearing bolt such that they can pivot, the contact mount has at least three supporting elements in order to support the bearing bolt radially (E 0 222 686 B1).\",\n \"The contact mount in this case includes a metal frame, which is formed from two side walls and from two or more bolts which connect the side walls.\",\n \"The metal frame is hinged via a coupling bolt on an insulating coupling element, which is used for coupling the contact mount to a switch drive.\",\n \"Two of the supporting elements, which are associated with the ends of the bearing bolt, are in this case formed by the side walls of the metal frame.\",\n \"In order to prevent undesirable radial bending of the bearing bolt for the contact levers with as little complexity in terms of additional material as possible, two intermediate bearings for this contact mount, which are arranged between adjacent contact levers in the axially central region of the bearing bolt, form additional supporting elements by being hinged on the coupling bolt.\",\n \"In this case, aperture openings are required for the contact mount, for the intermediate bearings to pass through to the coupling bolt.\",\n \"Supporting elements which are integrated in this way in addition to the two outer supporting elements in the contact mount must be positioned for installation of the bearing bolt, owing to their capability to pivot about the coupling bolt.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>Against the background of an electrical switch, an embodiment of the invention is based on an object of simplifying the production and installation of the contact mount.\",\n \"According to an embodiment of the invention, an object may be achieved in that at least three of the supporting elements are in the form of part of a molding which forms the contact mount and is produced integrally.\",\n \"For the purposes of an embodiment of the invention, the expression an integrally produced molding should be understood as being a part in which two or more functional elements are connected in the course of a molding process, such as a stamping, injection-compression molding, casting, injection molding, compression molding or sintering process, to form a single component which is assembled such that it cannot be disconnected for installation purposes.\",\n \"In the case of a refinement such as this, the three supporting elements are integrated rigidly in a predetermined position in the contact mount, as part of it.\",\n \"In this case, the three supporting elements are actually aligned with the axis of the bearing bolt during the production of the contact mount so that no tilting of the bearing bolt caused by tolerance discrepancies will in practice occur during operation of the switch.\",\n \"A bearing bolt which is supported in this way is also subject to only a small maximum amount of bending when high short-circuit or surge currents occur, and thus has a good capability to withstand high short-circuit and surge currents.\",\n \"The novel switching contact arrangement may have a large number of contact levers, which are each subject to an individual tolerance discrepancy from a given nominal size, and intermediate bearings, which are possibly likewise subject to an individual tolerance discrepancy from their nominal size, but which may be part of the contact levers, since the number of contact levers is subdivided into subsets.\",\n \"Each of these is arranged axially bounded between two adjacent supporting elements.\",\n \"This axial bounding of the subsets of contact levers limits any axial movement of the contact levers in one subset, owing to the current forces which act between them, to the axial section of the bearing bolt which is bounded by the respective supporting elements.\",\n \"The maximum amount of movement is not greater than the sum of all the individual tolerance discrepancies of the contact levers and of the intermediate bearings, which may be present, in this subset.\",\n \"This makes it possible to geometrically associate the contact levers with contact force springs such that their spring force is not reduced by bending or tilting.\",\n \"The geometrically accurate association between the contact force springs and the contact levers thus also contributes to increasing the capability of the switching contact arrangement to withstand short-circuit and surge currents.\",\n \"If the molding is at least partially in the form of a plastic molding, then there is no need for the coupling bolt to have an electrically insulating configuration.\",\n \"The mechanical strength of a plastic molding such as this can be increased by embedding at least one reinforcement element at least partially in the plastic molding.\",\n \"A thermosetting plastic which, for example, has fiber reinforcement is typically used for the plastic molding and a nonmagnetic steel, for example, is used for the reinforcement element.\",\n \"As an alternative to this, other pure plastics or, for example, plastics reinforced by ceramic or glass fibers can also be used for the plastic molding, and other metals or metal sheets can be used for the reinforcement element.\",\n \"One preferred refinement of the novel switching contact arrangement provides for at least one of the supporting elements to have a metal part which is at least partially embedded in the plastic molding.\",\n \"A metal part such as this may be part of the reinforcement element, thus at the same time increasing the mechanical strength of that part of the contact mount which forms the supporting element.\",\n \"If the metal part is in the form of a metal sheet, for example composed of nonmagnetic sheet steel, a first subregion of which, which has undercuts, is embedded in the plastic molding and a second subregion of which, which is provided with a hole for the bearing bolt, projects out of the plastic molding.\",\n \"This then advantageously allows the cross section of the supporting element to be reduced such that it is no broader than the distance between the contact levers that is required for separation of the contact levers and thus does not lead to any additional broadening of the contact mount.\",\n \"A further advantageous refinement of the novel switching contact arrangement provides for supporting elements which contain the metal parts to be at a distance from the coupling element in the axial direction of the bearing bolt if the contact mount is coupled to a switching shaft, which can be rotated using a switch drive, via a metallic coupling element.\",\n \"This makes it possible to avoid accidental energizing and problems relating to the insulation between the contact mount and the switch drive, in a simple manner.\",\n \"In this refinement, the entire available material depth of the contact mount transversely with respect to the bearing bolt can be used for the rigid embedding of a first subregion of a supporting element which is in the form of a metal sheet.\",\n \"If a holder for the shielding body is provided on at least one of the supporting elements for a contact mount which is equipped with the shielding body, then this provides additional support for the shielding body against the gas pressure which occurred during switching.\",\n \"In a refinement such as this, side mounting limbs, which rest on the contact mount, are designed to be smaller owing to the reduced load, or may possibly be omitted.\"\n ],\n [\n \"This application is the national phase under 35 U.S.C.\",\n \"§ 371 of PCT International Application No.\",\n \"PCT/DE02/01250 which has an International filing date of Mar.\",\n \"28, 2002, which designated the United States of America and which claims priority on German Patent Application numbers DE 101 17 844.1 filed Apr.\",\n \"4, 2001 and DE 201 18 493.1 filed Nov. 7, 2001, the entire contents of which are hereby incorporated herein by reference.\",\n \"FIELD OF THE INVENTION The invention generally relates to the field of electrical/power switches, and is preferably applicable to the design configuration of a rigid member which is used as a contact mount for a contact.\",\n \"BACKGROUND OF THE INVENTION In a known electrical switch of this type, in which two or more contact levers which form the contact are held on the contact mount by way of a bearing bolt such that they can pivot, the contact mount has at least three supporting elements in order to support the bearing bolt radially (E 0 222 686 B1).\",\n \"The contact mount in this case includes a metal frame, which is formed from two side walls and from two or more bolts which connect the side walls.\",\n \"The metal frame is hinged via a coupling bolt on an insulating coupling element, which is used for coupling the contact mount to a switch drive.\",\n \"Two of the supporting elements, which are associated with the ends of the bearing bolt, are in this case formed by the side walls of the metal frame.\",\n \"In order to prevent undesirable radial bending of the bearing bolt for the contact levers with as little complexity in terms of additional material as possible, two intermediate bearings for this contact mount, which are arranged between adjacent contact levers in the axially central region of the bearing bolt, form additional supporting elements by being hinged on the coupling bolt.\",\n \"In this case, aperture openings are required for the contact mount, for the intermediate bearings to pass through to the coupling bolt.\",\n \"Supporting elements which are integrated in this way in addition to the two outer supporting elements in the contact mount must be positioned for installation of the bearing bolt, owing to their capability to pivot about the coupling bolt.\",\n \"SUMMARY OF THE INVENTION Against the background of an electrical switch, an embodiment of the invention is based on an object of simplifying the production and installation of the contact mount.\",\n \"According to an embodiment of the invention, an object may be achieved in that at least three of the supporting elements are in the form of part of a molding which forms the contact mount and is produced integrally.\",\n \"For the purposes of an embodiment of the invention, the expression an integrally produced molding should be understood as being a part in which two or more functional elements are connected in the course of a molding process, such as a stamping, injection-compression molding, casting, injection molding, compression molding or sintering process, to form a single component which is assembled such that it cannot be disconnected for installation purposes.\",\n \"In the case of a refinement such as this, the three supporting elements are integrated rigidly in a predetermined position in the contact mount, as part of it.\",\n \"In this case, the three supporting elements are actually aligned with the axis of the bearing bolt during the production of the contact mount so that no tilting of the bearing bolt caused by tolerance discrepancies will in practice occur during operation of the switch.\",\n \"A bearing bolt which is supported in this way is also subject to only a small maximum amount of bending when high short-circuit or surge currents occur, and thus has a good capability to withstand high short-circuit and surge currents.\",\n \"The novel switching contact arrangement may have a large number of contact levers, which are each subject to an individual tolerance discrepancy from a given nominal size, and intermediate bearings, which are possibly likewise subject to an individual tolerance discrepancy from their nominal size, but which may be part of the contact levers, since the number of contact levers is subdivided into subsets.\",\n \"Each of these is arranged axially bounded between two adjacent supporting elements.\",\n \"This axial bounding of the subsets of contact levers limits any axial movement of the contact levers in one subset, owing to the current forces which act between them, to the axial section of the bearing bolt which is bounded by the respective supporting elements.\",\n \"The maximum amount of movement is not greater than the sum of all the individual tolerance discrepancies of the contact levers and of the intermediate bearings, which may be present, in this subset.\",\n \"This makes it possible to geometrically associate the contact levers with contact force springs such that their spring force is not reduced by bending or tilting.\",\n \"The geometrically accurate association between the contact force springs and the contact levers thus also contributes to increasing the capability of the switching contact arrangement to withstand short-circuit and surge currents.\",\n \"If the molding is at least partially in the form of a plastic molding, then there is no need for the coupling bolt to have an electrically insulating configuration.\",\n \"The mechanical strength of a plastic molding such as this can be increased by embedding at least one reinforcement element at least partially in the plastic molding.\",\n \"A thermosetting plastic which, for example, has fiber reinforcement is typically used for the plastic molding and a nonmagnetic steel, for example, is used for the reinforcement element.\",\n \"As an alternative to this, other pure plastics or, for example, plastics reinforced by ceramic or glass fibers can also be used for the plastic molding, and other metals or metal sheets can be used for the reinforcement element.\",\n \"One preferred refinement of the novel switching contact arrangement provides for at least one of the supporting elements to have a metal part which is at least partially embedded in the plastic molding.\",\n \"A metal part such as this may be part of the reinforcement element, thus at the same time increasing the mechanical strength of that part of the contact mount which forms the supporting element.\",\n \"If the metal part is in the form of a metal sheet, for example composed of nonmagnetic sheet steel, a first subregion of which, which has undercuts, is embedded in the plastic molding and a second subregion of which, which is provided with a hole for the bearing bolt, projects out of the plastic molding.\",\n \"This then advantageously allows the cross section of the supporting element to be reduced such that it is no broader than the distance between the contact levers that is required for separation of the contact levers and thus does not lead to any additional broadening of the contact mount.\",\n \"A further advantageous refinement of the novel switching contact arrangement provides for supporting elements which contain the metal parts to be at a distance from the coupling element in the axial direction of the bearing bolt if the contact mount is coupled to a switching shaft, which can be rotated using a switch drive, via a metallic coupling element.\",\n \"This makes it possible to avoid accidental energizing and problems relating to the insulation between the contact mount and the switch drive, in a simple manner.\",\n \"In this refinement, the entire available material depth of the contact mount transversely with respect to the bearing bolt can be used for the rigid embedding of a first subregion of a supporting element which is in the form of a metal sheet.\",\n \"If a holder for the shielding body is provided on at least one of the supporting elements for a contact mount which is equipped with the shielding body, then this provides additional support for the shielding body against the gas pressure which occurred during switching.\",\n \"In a refinement such as this, side mounting limbs, which rest on the contact mount, are designed to be smaller owing to the reduced load, or may possibly be omitted.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS The present invention will become more fully understood from the detailed description of preferred embodiments given hereinbelow and the accompanying drawings, which are given by way of illustration only and thus are not limitative of the present invention, wherein FIGS.\",\n \"1 to 9 show a number of exemplary embodiments of the novel switching contact arrangement and wherein: FIG.\",\n \"1 shows a schematic section illustration of a low-voltage circuit breaker with a switching contact arrangement which comprises a stationary contact assembly and a moving contact assembly, FIG.\",\n \"2 shows a moving contact assembly with a first embodiment of a contact mount which is at least partially in the form of a plastic molding, FIG.\",\n \"3 shows a reinforcement element, which may be embedded in the plastic molding of the contact mount shown in FIG.\",\n \"2, FIG.\",\n \"4 shows a second embodiment of a contact mount which is at least partially in the form of a plastic molding, and in which supporting elements in the form of a metal sheet are partially embedded in the plastic molding, FIG.\",\n \"5 shows a section illustration, transversely with respect to the direction of the bearing bolt, through the contact mount shown in FIG.\",\n \"4, FIG.\",\n \"6 shows a reinforcement element which may be embedded in the plastic molding of the contact mount in FIG.\",\n \"4, FIG.\",\n \"7 shows a section illustration, transversely with respect to the direction of the bearing bolt, through a contact mount with an embedded reinforcement element as shown in FIG.\",\n \"6, FIG.\",\n \"8 shows a further section illustration through the contact mount illustrated in FIG.\",\n \"4 along the line A-A in FIG.\",\n \"5, and FIG.\",\n \"9 shows the contact lever mount as shown in FIG.\",\n \"4 with a shielding body.\",\n \"DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The electrical switch which is shown in FIG.\",\n \"1 and is used in low-voltage power supply systems for voltage ranges up to about 1 000 V has a switch pole enclosure 1 in which switching chambers 2 are formed alongside one another, depending on the number of poles required.\",\n \"A switching shaft 4, which can be rotated by way of a switch drive 3, is used to operate the switching contact arrangement jointly, each of which includes a stationary contact assembly 5 and a moving contact assembly 6.For this purpose, two levers 7 which project radially from the switching shaft 4 (see FIG.\",\n \"8) are coupled to a metallic coupling element which is hinged on the moving contact assembly 6.The contact assemblies 5, 6 are connected in a known manner to externally accessible connecting rails 9, 10.Two exemplary embodiments 6a and 6b of the moving contact assembly 6 will be explained in the following text with reference to FIGS.\",\n \"2 and 3 and, respectively, FIGS.\",\n \"4 to 9.As can be seen in more detail from FIG.\",\n \"2, the moving contact assembly 6a has a contact mount 11 which has a plastic molding 12 in the form of an integrally produced molding, which was formed in the course of a stamping process, with a reinforcement element as shown in FIG.\",\n \"3 being embedded in it.\",\n \"The contact mount is mounted in the enclosure 1 (see FIG.\",\n \"1) such that it can pivot, and can be moved via the switching shaft 4 and by use of the switch drive 3, of which FIG.\",\n \"1 shows only one drive run 15 that is supported on a spring stalk 14 relative to the stationary contact assembly 5 to a connected position and to a disconnected position.\",\n \"Two or more contact levers 16, 17, which are arranged parallel to one another, on the contact mount 11 can pivot relative to the contact mount 11 about a bearing bolt 18.Contact force springs 19 (see FIG.\",\n \"1) ensure that the contact levers 16, 17 are prestressed in the direction of the stationary contact assembly 5.Flexible conductors 20 in the form of braids or strips are used for connecting the contact levers 16, 17 to the lower connecting rail 10 in such a way as to guarantee that the contact levers 16, 17 and the contact mount 11 can move without any impediment during the switching movements.\",\n \"The number of contact levers 16, 17 which are fitted to the contact mount 11 depends on the magnitude of the current which the circuit breaker is intended to carry during operation.\",\n \"As can be seen from FIG.\",\n \"2, of the total of 22 contact levers that are provided, 8 contact levers 16 are designed to be shorter and have only one contact area 21, which have no leading contact area 22 and no arcing horn 23 in the same way as the other contact levers 17.During operation, all the contact levers are held between side pieces 24a, 24b of the contact mount 11, which point transversely with respect to the bearing bolt 18.These side pieces 24a, 24b, which are provided with holding openings 25a, 25b for the bearing bolt 18 form a first and a second supporting element for the ends of the bearing bolt, via which the bearing bolt is positioned axially and is supported radially.\",\n \"A part 29a or 29b of the reinforcement element 13 (see FIG.\",\n \"3) can extend in each of these side pieces, and has an aperture 27a or 27b, respectively, for the bearing bolt.\",\n \"In the downward direction, the side pieces 24a, 24b merge into bearing arms 26 for the contact mount 11.The relatively large width of the switching contact arrangement indicates that the section of the bearing bolt which runs between the two side supporting elements 24a, 24b is subjected to a relatively severe bending load when further forces in addition to the forces of the contact force springs 19 are caused by a heavy current, such as a short-circuit or surge current, when the switching contact arrangement is in the closed state.\",\n \"Bending of this section of the bearing bolt is prevented by way of an additional, third supporting element, which supports the bearing bolt axially in the center.\",\n \"This third supporting element is formed by a contact mount rib 28, which is provided with a holding opening 25c (which cannot be seen in the figure) for the bearing bolt 18 and points transversely with respect to the bearing bolt, with a metal part 29c (which is completely embedded in the plastic molding 12 and has an aperture 27c for the bearing bolt) extending in the rib 28 and being part of the reinforcement element 13 (see FIG.\",\n \"3).\",\n \"Of the second exemplary embodiment 6b of the moving contact group, FIG.\",\n \"4 shows only a second embodiment 30 of the contact mount.\",\n \"In this contact mount 30, two supporting elements, which are in the form of metal sheets 31, are used to radially support that section of the bearing bolt which runs between two supporting elements that are in the form of side pieces 32a, 32b.\",\n \"A first subregion 33 of the two central supporting elements 31 is embedded in the plastic molding 34 of the contact mount 30, and together with a second subregion 35 of the two central supporting elements 31, projects out of the plastic molding.\",\n \"As can be seen in FIG.\",\n \"5, the metal sheets 31 have undercuts 36 in the first subregion 33 which is embedded in the plastic molding, and these are used to anchor the respective metal sheet in the plastic molding securely even when the bearing bolt is subjected to a high bending load.\",\n \"The second subregion 35, which is provided with a hole 38 for the bearing bolt 37 to pass through, also has a recess 39, which is used to hold a shielding body 40 that is not shown in any more detail in FIG.\",\n \"9.As is shown in FIG.\",\n \"7, two reinforcement elements 41, 42 may be embedded in the plastic molding of the contact mount, and one of these is illustrated in FIG.\",\n \"6.As can be seen from FIG.\",\n \"8, the two metal sheets 31 are at a distance from the metallic coupling element 8 (see FIG.\",\n \"1) in the axial direction, in order to avoid accidental energizing between the bearing bolt (which is at a low-voltage potential) of the contact levers and the metallic coupling element 8 (which is at ground potential), and thus the switch drive.\",\n \"As can be seen from FIG.\",\n \"9, tongues 43 are integrally formed on the shielding body 40, which protects the pivoting area of the contact mount 30 and further switch parts (which are not shown in any more detail but are arranged underneath the contact areas 21, 22 (see FIG.\",\n \"1)) against erosion products that fall out and against condenser switching gases, and these tongues 43 engage in the recesses 39 which are provided on the metal sheets (see also FIG.\",\n \"5).\",\n \"In consequence, the shielding body 40 is supported against the gas pressure which occurs during switching processes, in such a way that its mounting limbs 44a, 44b, which are held in the side on the contact mount, are less severely loaded.\",\n \"Exemplary embodiments being thus described, it will be obvious that the same may be varied in many ways.\",\n \"Such variations are not to be regarded as a departure from the spirit and scope of the present invention, and all such modifications as would be obvious to one skilled in the art are intended to be included within the scope of the following claims.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473972"}}},{"rowIdx":774,"cells":{"text":{"kind":"list like","value":[["Method for determining effective parathormone activity in a sample","Method for determining the active parathyroid hormone content in a sample, wherein the component of parathyroid hormone polypeptide chains which are oxidized at one or more sites near the PTH receptor binding structure having the amino acids 15 to 22 are specifically excluded from the content determination.","Oxidation of the parathyroid hormone occurs in particular in dialysis patients, their blood plasma being exposed to an oxidation stress in the dialysis.","The invention is based on the use of antibodies which bind specific conformation epitopes of the oxidised or reduced parathyroid hormone and its fragments.","Further, there is provided a test system for the quantitative determination of parathyroid hormone and active fragments hereof in a sample."],["1.Method for determining the effective parathyroid hormone content in a sample of a body fluid, characterized in that there are specifically excluded from the content determination intact parathyroid hormone polypeptide chains and N-terminal intact fragments hereof which are oxidized at one or more sites near the PTH receptor binding structure having the amino acids 15 to 22.2.Method according to claim 1, wherein the methionine at position 8 or 18 of the parathyroid hormone polypeptide chain, or at both positions, is oxidized.","3.Method according to claim 1 or 2, wherein antibodies are employed which bind to a PTH epitope which is formed of at least one oxidized methionine group at position 8 and/or 18.4.Method according to any of claims 1 to 3, wherein antibodies are employed which bind specific parathyroid hormone polypeptide chains or fragments hereof which are oxidized at methionine 8 and/or 18.5.Method according to any of claims 1 to 4, wherein antibodies are employed which bind specific conformation epitopes of the oxidized or native, reduced parathyroid hormone.","6.Method according to any of claims 3 to 5, wherein the antibody employed is purified or selected by means of a binding to parathyroid hormone which is oxidized at position 8 and/or 18.7.Method according to any of claims 1 to 6, wherein intact parathyroid hormone and fragments hereof are oxidized and/or reduced and then the total content of intact parathyroid hormone and its active fragments is determined.","8.Method according to claim 7, whereby the total content of oxidized and reduced intact parathyroid hormone and its fragments is determined.","9.Method according to any preceding claim, wherein the contents of oxidized intact parathyroid hormone and fragments hereof are separately determined.","10.Method according to any preceding claim, whereby antibodies according to claim 3 or 4 are put to use for masking of oxidized parathyroid hormone and its fragments, so that exclusively reduced, biologically active parathyroid hormone and its fragments are determined.","11.Method according to any preceding, including the following steps: treating the sample with an antibody which recognizes an epitope which is formed from the ends of the N-terminal amino acids 1 to 3 of the parathyroid hormone; treating the sample with an antibody which recognizes an epitope in the region of the receptor binding site between amino acids 15 and 22 of human parathyroid hormone, and determination of the molecules which are recognized by the two antibodies.","12.Method according to claim 11, wherein the binding of the two antibodies is effected in the presence of 0.05 to 0.1 weight % of a mild detergent such as Tween™ or Triton™X-100.13.Method according to claim 11 or 12, wherein the sample is treated with antibodies against oxidized parathyroid hormone in accordance with claim 3 or 4, so that antibodies which bind in the region of the receptor binding site of the parathyroid hormone cannot bind to the oxidized parathyroid hormone chain.","14.Method according to any preceding claim, wherein the antibodies for masking of oxidized parathyroid hormone and fragments hereof are of a type different from the antibodies for building the sandwich complex.","15.Method according to any preceding claim, wherein at least one of the antibodies carries a marking selected from a fluorescing or chemiluminescing group or an enzyme for catalysis of a detection reaction.","16.Method according to any preceding claim, wherein one of the antibodies is a monoclonal antibody marked with a ruthenium complex and recognizes an epitope in the region of the amino acids 15 to 22 or 26 to 32.17.Method according to any preceding claim, whereby one of the antibodies is biotinylated.","18.Method according to any preceding claim, whereby antibodies are employed which bind to the parathyroid hormone polypeptide chain with amino acids 4 to 14 or 15 to 35v39.19.Test system for the quantative determination of parathyroid hormone and fragments-hereof in a sample, characterized by antibodies which bind to or near to the PTH receptor binding structure of the parathyroid hormone, and by antibodies which bind the epitopes of the parathyroid hormone having oxidized methionine 8 and/or 18.20.Test system according to claim 19, wherein the polyclonal antibody against the N-terminal fragment 1 to 37 of the parathyroid hormone is purified by means of affinity chromatography of antibodies which bind to methionine 8 and/or 18 of the oxidized parathyroid hormone fragment.","21.Test system according to claim 19 or 20, wherein there are determined intact parathyroid hormone and N-terminal fragments hereof having the amino acids 1 to 3 in the reduced structure.","22.Diagnosis of oxidation stress in dialysis patients, characterized by the employment of an in vitro test for the quantitative determination of parathyroid hormone and its active fragments in accordance with any preceding claim."],["The invention relates to an immunological method for determining the effective concentration of parathyroid hormone contained in a sample.","The invention relates in particular to a test kit for the determination of the content of active parathyroid hormone and its biologically active fragments for diagnosis, etiology and treatment of disturbances of the calcium metabolism, osteopathies and hyper- or hypo-parathyroidism.","The parathyroid hormone (PTH) is formed in the parathyroid gland (Glandulae parathyroideae) and secreted into the blood circulation.","In the intact form it consists of a single polypeptide chain having 84 amino acids and has a molecular weight of ca.","9500 Dalton (see SWISS-PROT: P01270, PTHY-HUMAN).","Together with Vitamin-D and calcitonin it brings about the mobilization of calcium and phosphate out of the bone skeleton and increases the uptake of calcium in the intestines and the excretion of phosphate via the kidneys.","The effective PTH activity in plasma or serum is thus an important diagnostic parameter and necessary for determining 1) presence and degree of hyper- or hypo- parathyroidism, 2) quantification of osteoblast activity, 3) quantification of osteoclast activity 4) monitoring of treatment with Vitamin-D and active Vitamin-D metabolites, 5) estimation of presence of aluminium, 6) estimation of a possible oestrogen deficiency in post-menopausal dialysis patients, 6) the necessary steroid or cyclosporin dosage after kidney transplantations, 7) the need for treatment and the prevention of pathological bone marrow changes, uraemic conditions and chronic kidney failure.","In the state of the art the determination of PTH is effected by means of the detection of two separated epitopes on the PTH polypeptide chain.","With the introduction of specific highly sensitive immunoassays for determining the quantity of intact PTH chains in human serum there appeared to be answered all diagnostic questions in the field of calcium homeostasis, bone transformation and the responsible controlling organs such as the parathyroid gland.","However, the values so determined often do not correspond to the actually present PTH activity in the plasma or serum sample, because they often provide PTH contents which are not reconcilable with the symptoms of the patients.","In the case of patients with uraemic conditions with normal bone transformation, the levels of intact PTH in serum lie mostly higher by the factor 2.5 than with patients with healthy kidneys (pathological limit in the case of patients with healthy kidneys: 65 μg PTH/L; for patients having uraemic conditions: 165 μg PTH/L serum) Further, uraemic patients with comparably high PTH values often manifest significant differences in bone transformation (Slatopolsky E et al.","(2000), Kidney Int., 58, 753-761).","Thus these patients often have in the serum eight to ten times increased PTH concentrations, but low normal values for bone specific alkaline phosphatase (ostase).","They are thus free from symptoms of an excessive PTH activity.","As possible causes there have been discussed either systematic errors in the determinations or a PTH resistance of the osteoblasts, for example due to genetically reduced expression of PTH receptor.","Further, the determination of the effective PTH content in serum or plasma is made more difficult in that the hormone is broken down within minutes, in the circulation or the liver, into active and inactive fragments.","Some of these fragments have an activity comparable with intact PTH (see EP-A 0 349 545; Schmidt-Gayk et al.","(1999) Osteologie forum, 5, 48-58), on the other hand others work antagonistically.","Thus, one knows that the fragments hPTH (3-34) and hPTH (7-34) can inhibit the effects of PTH (Suva et al.","(1987) Science, 237, 893ff; EP 0 451 867).","Moreover, “non-(1-84)” large PTH fragments have been suspected to lie behind the erroneous determinations (LePage R. et al.","(1998) Clin.","Chem., 44, 805-809).","The designation “Large PTH fragment” is used generally for fragments which contain at least the polypeptide chain with the amino acids 39-84 and epitopes of the N-terminal fragments hPTH (1-33), hPTH (1-34), hPTH (1-37) or hPTH (1-38) but from which the N-terminal amino acids 1 to 3 are missing.","WO 96/10041 teaches that after loss of the N-terminal amino acids serine and valine the PTH fragments are no longer active.","The characterising of various immunological tests with synthetic fragments of the type hPTH (7-84) showed that these as a rule also determine the inactive large PTH (7-84) fragment (John M R et al.","(1999), J. Clin.","Endocrinol.","Metab., 84.4287-4290; Gao P et al.","2000, Poster M455, ASBMR 22nd Annual Meeting; Roth H J et al.","(2000), Poster P1288; 11th International Congress of Endocrinology, Sydney).","The co-determination of inactive large PTH fragments was thus made responsible for the discrepancy between measured PTH concentration and clinical findings.","It was suspected that intact PTH molecules compete with the large PTH fragments for the binding sites on the PTH receptor and that depending upon the concentration of CAP (Cyclase activating PTH—biologically active) or CIP (Cyclase Inhibiting PTH—biologically inactive, receptor blocking) different cell activations occurred.","It is object of the invention to make available a method for the quantative determination of the effective PTH content of a sample.","This object is achieved by means of the method according to claim 1.Further advantageous embodiments of the method are indicated in the subclaims.","The method in accordance with the invention for quantitative determination of the PTH content in a sample is characterized in that intact PTH polypeptide chains and N-terminal intact fragments hereof having the PTH receptor binding structure (15 to 22), which are oxidised at one or more sites neighbouring the PTH receptor binding structure (15 to 22), are excluded from the content determination.","In other words there are excluded from the determination of the PTH activity or the effective PTH content all PTH polypeptide chains and fragments hereof which are oxidised at methionine-8 or -18 or at both positions.","This is effected through the use of antibodies which bind to a PTH epitope which is formed from at least one oxidised methionine group at position 8 and/or 18, or of antibodies which bind specifically PTH polypeptide chains or fragments hereof which are oxidised at methionine 8 and/or 18, or antibodies which can distinguish between specific conformation epitopes of the oxidised and of the “native” non-oxidised parathyroid hormone.","It is this ensured that there are excluded from the determination: inactive large PTH fragments of the types hPTH (7-84) or hPTH (3-84), oxidised intact hPTH (1-84) and oxidised N-terminal intact oxidised hPTH (1-34) fragments of the type ox-hPTH (1-34) or ox-hPTH (1-37), which contain the receptor binding structure PTH (15-22).","In accordance with the invention, the antibodies are obtainable via a purification or selection on parathyroid hormone which is oxidised at position 8 and/or 18.The antibodies can be obtained through immunization with synthetic or specifically oxidised PTH fragments (Logue F C et al.","(1991), Ann.","Clin.","Biochem., 28 160-166; Journal of Immunological Methods, 39, 159).","For example, synthetic PTH (1-34) can be oxidised at the methionines 8 and 18 through the addition of H2O2 and put to use for the immunization.","Particularly preferred are monoclonal antibodies against methionine-8, which apparently is oxidized first.","In an embodiment of the method in accordance with the invention, the parathyroid hormone and its potentially active fragments are chemically post-oxidized and/or post-reduced and the content of intact parathyroid hormone and its active fragments is determined by a double determination (before and after).","In a further embodiment, there is separately and additionally determined the content of oxidised intact parathyroid hormone and fragments hereof in the sample and calculated via the simultaneous determination of total PTH and the content of non-oxidized PTH and its biologically active fragments.","In a particularly preferred embodiment of the method there are added antibodies against parathyroid hormone oxidized at position 8 and/or 18, for masking the oxidized parathyroid hormone, so that exclusively non-oxidized or biologically active parathyroid hormone and its fragments are bound by primary and secondary antibodies.","The method in accordance with the invention then includes the following steps: treating the sample with an antibody which recognizes an epitope in which parathyroid hormone oxidized at position 8 and/or 18 is involved; treatment of the sample with an antibody which recognizes an epitope which is formed from the N-terminal amino acids 1 to 3 of the parathyroid hormone; treatment of the sample with an antibody which recognizes an epitope in the region of the receptor binding structure (amino acids 15 to 22) of human parathyroid hormone and which cannot bond when the antibody has already bonded in the region of the oxidized methionine 8 and/or 18, and determination of the number of the molecules which are recognized by the two antibodies.","Preferably, here the binding of the two antibodies takes place in the presence of the 0.05 to 0.1 weight percent of a mild detergent such as Tween™20 or Triton™X-100.In accordance with the invention, in a particularly preferred embodiment, the sample is thus first treated with antibodies against oxidized parathyroid hormone, so that antibodies which bind in the region of receptor binding site of the parathyroid hormone cannot bind to the oxidized PTH polypeptide chain.","The person skilled in the art will recognize that the antibody for masking of oxidized parathyroid hormone and fragments hereof should better, be of a different type than the antibody for forming or detecting the sandwich complex.","The antibodies employed as captors and tracers, may, as the skilled person knows, also be functionally exchanged.","In a further embodiment at least one of the antibodies carries a marking, selected from a fluorescing or chemiluminescing group or an enzyme for catalysing a detection reaction.","Preferably one of the antibodies is a monoclonal antibody marked with a ruthenium complex, which recognizes an epitope in the region of the amino acids 15 to 22 or 26 to 32.One of the antibodies may also be biotinylated and bonded via streptavidin to a solid phase.","In the method there can further be employed antibodies which bind to the PTH polypeptide chain with amino acids 4 to 14 or 15 to 35v39.An antibody which recognizes an epitope in the range of amino acids 15 to 22 is particularly preferred since on the one hand this epitope lies in the range of the receptor binding structure, on the other hand such an antibody can distinguish, due to the methionine 18, between oxidized and native hPTH.","The test system in accordance with the invention for the quantitive determination of parathyroid hormone and fragments hereof in a sample includes in one embodiment antibodies (captor antibodies) bound to a phase, which antibodies bind in or near the PTH receptor binding structure of the parathyroid hormone and—for masking—free (unbound) antibodies, which bind the epitopes of the parathyroid hormone having oxidized methionine 8 and/or 18.The antibodies can be purified through affinity chromatography, for example via their binding to methionine 8 and/or 18 of the oxidized parathyroid hormone.","As tracer antibodies there are employed antibodies against the N-terminal amino acids 1 to 3 of the parathyroid hormone (in the reduced structure).","Captor and tracer antibodies may also be exchanged.","The invention further relates to the diagnosis of oxidation stress in dialysis patients, whereby an in vitro test for the quantitative determination of parathyroid hormone and it active fragments in accordance with the invention is put to use.","Even though one knew from experiments with synthetic PTH fragments that oxidized parathyroid hormone or oxidized parathyroid hormone fragments are biologically inactive, one did not imagine that this oxidation could assume physiologically relevant levels and prevent the determination of the true physiological PTH activity in a sample.","Thus in some conventional test methods there were added reduction means for stabilizing the antibodies in the sample buffer, in others a subsequent partial oxidation of the PTH molecule was tolerated.","The invention also relates in an embodiment to a method for a diagnosis and determination of the extent of a hypo- or hyper-parathyroidism, the etiology of calcium metabolism disturbances, osteopathies, kidney failures and conditions which arise from a disturbed homeostasis of the calcium and phosphate contents of the blood.","Further, in accordance with the invention the direct production of N-terminal PTH fragments, so far as this arises, is also detected.","This means that the PTH diagnosis in accordance with the invention detects along with intact active parathyroid hormone also directly produced, biologically active PTH fragments.","Oxygen is essential for aerobic life but at the same time is the most important precursor molecule of free reactive oxygen species (ROS) which through their reaction with proteins, lipids and nucleic acids damage the cells and through the oxidation of reactive amino acids can alter proteins in their structure.","Many publications have been concerned with the effect of oxidation stress and the influence of “advanced glycation end products” on the pathogenesis of complications in the case of chronically kidney insufficient patients (Martin-Mateo M C et al.","(1999), Ren.","Fail., 21, 155-167; Hasselwander O et al.","(1998), Free Radic.","Res., 29, 1-11; Zoccali C et al.","(2000), Nephrol.","Dial.","Transplant., 15, Suppl.2; Canaud B et al.","(1999), Blood Purif., 17, 99-106).","Dialysis patients are in comparison to normal persons much more strongly affected by oxidative stress.","Various working groups have investigated oxidized parathyroid hormone and its biological activity (Alexiewicz L M et al.","(1990), J.","Am.","Soc.","Nephrol., 1, 236-244; Zull J E et al.","(1990), J. Biol.","Chem., 265, 5671-5676; Pitts T O et al.","(1988), Miner.","Electrolyte Metab., 15, 267-275; Horiuchi N. (1988), J.","Bone Miner.","Res., 3, 353-358; Frelinger A L et al.","(1986) Arch.","Biochem.","Biophys., 244, 641-649; Galceran T et al.","(1984) Endocrinology 115, 2375-2378; Frelinger A L et al.","(1984), J. Biol.","Chem., 259, 5507-5513; O'Riordan J L H et al.","(1974), J.","Endocr., 63, 117-124; Logue F C et al.","(1991), Ann.","Clin.","Biochem., 28, 160-166; Logue F C (1991b), J. Immun.","Meth., 39, 159).","Oxidation stress in dialysis patients and its consequence for PTH diagnosis have, however, not so far been investigated and recognized.","It has now been found that the conventional methods of determination, in each case depending on the test kit and antibodies, detect both bioactive (native or reduced) intact PTH and if applicable its fragments and also oxidized PTH and if applicable its fragments, and thus falsely determine the effective level of parathyroid hormone.","Further, it has been found that the postulated large PTH fragments, insofar as they arise at all, have no biological significance: rather that the discrepancy between measured PTH content and medical determination arises at least partly from the oxidation stress to which the dialysis patient is subject.","The effective PTH content in a plasma or serum sample is namely dependent upon the oxidation stress and the oxidation stress is at the same time a cause of pathological conditions.","The invention relates further to a method in which a first captor antibody is bonded to a solid phase.","The second tracer antibody may carry a marker, e.g.","a fluorescent or chemiluminescent group, or may be conjugated with an enzyme such as alkaline phosphatase or peroxidase.","The method is effected in accordance with the invention in a per se known immunoassay, preferably in an ELISA, ECLIA, IEMA, IRMA, ILMA or LIA.","The binding of the two antibodies to the PTH is preferably effected in the presence of 0.05 to 0.1 weight percent of a mild detergent such as Tween™20 or Triton™X-100 so that a folding of the N-terminal epitope PTH (1-3) to the PTH receptor binding structure is prevented, or in order to keep the N-terminal epitope PTH (1-3) accessible for binding with the antibody.","Since for the biological activity and the folding of the PTH both the receptor binding structure and also the amino-terminal epitope PTH (1-3) are necessary, these two structures probably interact with one another.","Through the presence of a mild detergent, in the above-mentioned embodiment with antibodies against hPTH (1-3), the sensitivity and accuracy of the assay can be significantly improved.","In an embodiment of the method a known aliquot of the sample is further treated with antibodies which bind to the PTH sequences between amino acids 4 to 14 and/or 15 to 37.The number of molecules in a sample which are bound by antibodies against the epitope PTH (1-3) and the receptor binding structure of the PTH, and the number of molecules which are bound by the antibodies against the PTH regions 4 to 14 and 15 to 37, are then placed in relationship one to another and the biologically effective PTH activity determined.","The invention further relates to a diagnosis system for the determination of the PTH activity in a sample, which system is characterized by antibodies which recognize specifically the N-terminal epitope with the amino acids 1 to 3 and antibodies which bind to the region of the PTH receptor binding structure.","In a further embodiment the diagnosis system further has antibodies which bind in the region between amino acids 4 to 14.In a third embodiment the system includes antibodies which bind in the section between amino acids 24 and 37 of the parathyroid hormone or on the middle (53 to 86) or the C-terminal (53 to 84) section of the PTH polypeptide chain.","In contrast to earlier immunoassays, the method in accordance with the invention takes into account the PTH activity also of the non-intact PTH fragments and that apparently intact PTH is biologically inactive when the last or last two amino terminal amino acids of the parathyroid hormone are missing or when one or both methionines at position 8 and 18 of the N-terminal fragments are oxidized.","PTH fragments which have neither a receptor binding structure with the amino acids 15 to 22 nor an intact N-terminal epitope PTH (1-3) have no agonistic or antagonistic activity.","Conventional immunological tests for the quantative determination of intact parathyroid hormone or hPTH (1-37) yield false activity values if a physiological activity is concluded upon only from the presence of two epitopes.","Conventional methods do not take into account that the correct folding or the binding to the PTH receptor is dependent upon an intact amino-terminus with the amino acids serine and valine and that the methionine groups at position 8 and 18 near the PTH receptor binding structure must not be oxidized.","The invention rests in particular on the discovery that the oxidation of PTH at the above-mentioned positions, in particular in the case of dialysis patients, can be considerable and of diagnostic significance.","The fragments hPTH (7-84), hPTH (3-84) or hPTH (4-37), so far as they are present, determined in conventional tests as active PTH have no or no measurable antagonistic activity.","Test systems on the basis of antibodies against the middle section (53-68) or the C-terminal (53-84) section overlook the fact that some patients directly produce biologically active PTH fragments of the type hPTH (1-37), hPTH(1-32˜36) or hPTH(1-38).","Immunological methods on the basis of antibodies against hPTH(1-3) or hPTH(1-6), or against peptides from the hPTH(1-37) fragments, also reflect false activities since there are determined not only the biologically active (reduced) structure units.","With polyclonal antibodies or antisera against the N-terminal peptide having the amino acids 1 to 5 or 6 in part even antagonistically effective fragments are recognized as active PTH fragments.","The method in accordance with the invention thus makes available to the doctor a PTH activity value which indicates the physiological effects of the native (reduced) parathyroid hormone and its active fragments.","It recognizes also patients hyperparathyroidism who have normal or reduced contents of intact hPTH(1-84) in the serum and directly secrete PTH fragments such as hPTH(1-37) and hPTH(1-38) into the bloodstream.","The method in accordance with the invention for determining the activity of PTH and its fragments in a sample can be realized as EIA, ELISA, RIA, IRMA, LIA or ICLIA, ILMA, FIA or IFMA, as a manual test system or preferably in a version adapted to automatic systems, in liquid phase or solid phase techniques.","Further advantages, features and embodiments of the invention are provided through the following examples and the accompanying drawings.","There is shown: FIG.","1 a diagram with results (test system: Elecsys™2010 of Roche Diagnostics GmbH, Mannheim) of spiking experiments on PTH standard; FIG.","2 a diagram with results (Elecsys™2010 of Roche Diagnostics) of spiking experiments on pool plasmas; FIG.","3 a diagram with results (test system: PTH-IRMA of the Nichols Institute, U.S.A.) of spiking experiments on PTH standard; FIG.","4 a diagram with results (PTH-IRMA of the Nichols Institute) of spiking experiments on pool plasmas; FIG.","5 a diagram with measured values (Roche Elecsys™2010 of the plasma samples after oxidation with H2O2; FIG.","6 a diagram with percentage measurement values (Roche Elecsys™2010) of the plasma samples after oxidation with H2O2; FIG.","7 a diagram with measurement values (PTH-IRMA of the Nichols Institute) of the plasma samples after oxidation with H2O2; FIG.","8 a diagram with percentage measurement values (PTH-IRMA of the Nichols Institute) of the plasma samples after oxidation with H2O2; FIG.","9 a diagram having measurement values (PTH-IRMA of the Nichols Institute) after oxidation and reverse reduction of the samples; FIG.","10 a diagram with percentage measurement values (PTH-IRMA of the Nichols Institute) after oxidation and reverse reduction of the samples; FIG.","11 a diagram with percentage measurement values (PTH-IRMA of the Nichols Institute) after oxidation and direct reduction of the samples; FIG.","12 a diagram with percentage measurement values (PTH-IRMA of the Nichols Institute) of a plasma sample with and without oxidization and after addition of monoclonal antibody against oxidized parathyroid hormone; FIG.","13 a diagram with percentage measurement values from a plasma sample (Roche Elecsys™) with and without oxidization and after addition of monoclonal antibody against oxidized parathyroid hormone.","EXAMPLES Example 1 Biological Effect of Large PTH Fragments It was investigated whether the large PTH fragments, postulated in the state of the art, occur in human serum and have an antagonistic effect on the PTH receptor.","The immunoassays employed were based upon antibodies which do not cross-react with synthetic hPTH(7-84) (Bachem AG), so that the measurement values should indicate the true concentration of intact active hPTH in human serum.","These values were compared with parameters for bone transformation.","The production of the large PTH fragments is, in accordance with the state of the art, regulated via a feedback mechanism and in the case of increased or reduced need the stimulation or suppression of the parathyroid gland leads to an adequate expression.","At the present time there are known two test systems for the quantative determination of PTH which have no clinically relevant cross-reactivity with the synthetic large PTH fragment hPTH(7-84).","These are the Elecsys™2010 PTH immunoassay of Roche Diagnostics GmbH, Mannheim and the CAP-PTH-IRMA of the company Scantibodies.","The electrochemiluminescence immunoassay (Elecsys™ of Roche Diagnostics GmbH, Mannheim) is based upon a monoclonal captor antibody against an N-terminal PTH epitope corresponding to the amino acids 26-32 and a ruthenium complex marked tracer antibody against a C-terminal hPTH epitope corresponding to the amino acids 55-64.The employed synthetic large hPTH(7-84) fragment (Bachem AG) is not recognized by the captor antibody of the Elecsys™2010 PTH immunoassay, so that it must recognize a tertiary folding structure or is a so-called conformation antibody.","Probably, the large hPTH(7-84) fragment employed (Gao P et al.","2000, Poster M455, ASBMR 22nd Annual Meeting, Roth H J et al.","(2000), Poster P1288; 11th International Congress of Endocrinology, Sydney), as the present results make evident, was oxidized in the course of synthesis or storage.","The captor antibody of the Elecsys™2010 PTH immunoassay probably does not bind only oxidized hPTH(7-84), but does bind reduced PTH(7-84).","The CAP-PTH-IRMA of the company Scantibodies employs polyclonal antibodies against the N-terminal hPTH(1-6) which could not bind the large PTH(7-84) fragments.","Along with the determination of intact PTH using various immunoassays (Nichols IRMA, Roche Elecsys™2010, DUO-PTH Scantibodies, PTH-Immundiagnostik) there were also determined the bone marker BAP (ostase) as a formation marker and TRAP5b as resorption marker.","The results are indicated in the following Table 1.TABLE 1 Correlation of bone growth and reduction markers with PTH measurement values Dependent Independent Correlation variable Y variable X coefficient/n= BAP (Ostase) PTH CAP Scan.","0.484 n = 96 BAP (Ostase) PTH Elecsys2010 0.474 n = 96 BAP (Ostase) PTH Nichols 0.485 n = 96 TRAP5b PTH CAP Scan 0.511 n = 95 TRAP5b PTH Elecsys2010 0.510 n = 95 TRAP5b PTH Nichols 0.524 n = 95 PTH CAP Scan PTH Elecsys2010 0.966 n = 103 PTH CAP Scan PTH Nichols 0.979 n = 103 The correlations of the values of the growth and reduction markers and the PTH contents in the serum show no significantly greater correlations for the more specific tests (“no cross reactivity to large PTH fragments”) in comparison to the IRMA of the company Nichols Institute.","Amongst one another the correlation of the individual PTH assays is, as has also been shown in other studies, excellent.","This shows that none of the known test systems provide satisfactory results.","Example 2 Oxidation and Reduction Experiments It was investigated whether and to what extent the Elecsys™2010 PTH test and the Nichols Intact PTH test recognize native or oxidized PTH.","The Nichols Intact PTH Assay (Nichols Institute, San Juan Capistrano, Calif., U.S.A) is a double binding immunoradiometry assay (IRMA) with two polyclonal goat antibodies.","The captor antibody, immobilized in plastic beads, binds only in the middle or C-terminal section of the hPTH (anti-hPTH(39-89)).","The second detection antibody, marked with 125Iodine, recognizes an epitope in the N-terminal section of the PTH (anti-hPTH(1-34)).","The sample was incubated at the same time with the antibody coated beads and the 125Iodine marked detection antibody overnight (12±2h) at room temperature.","After the incubation, the beads were washed and the radioactivity adhering to the solid phase measured in a gamma counter (LKB1277 Gamma Master from Perkin-Elmer).","The dose measurement curve (MultiCalc Software) between bound radioactivity and concentration in the sample was determined strictly in accordance with the manufacturer's instructions via a standard measured at the same time.","The interassay variance in the normal and increased region was less than 10% CV.","The Roche Elecsys™ PTH assay is an electrochemiluminescence immunoassay (ECLIA) on the fully automatic immunoanalyzer Elecsys R2010.The epitope recognized by the biotinylated captor antibody contains the PTH amino acids 26 to 32 and the monoclonal ruthenium detection antibody recognizes, in accordance with the information from the manufacturer, an epitope in the region of the PTH amino acids 55 to 64.Sample and antibodies were incubated together for 9 minutes at 37° C., Streptavidin coated paramagnetic beads introduced to the incubation mixture, and finally incubated for further 9 minutes.","After magnetic separation of the bonded conjugates from the free conjugates on an electrode surface the luminescence was determined and the end PTH concentration determined via a recalibrated master curve.","The interassay variance in the lower and upper normal region was less than 7% CV and in the high concentration region 5% CV.","For the oxidation and reduction experiments standards and pool plasmas from dialysis patients having hPTH (1-37) fragments of different concentrations and oxidation states were treated.","All samples received EDTA as anticoagulant.","The measurement of the native, unaltered initial samples (reference parameter) and of the treated pool plasmas was effected at the same time.","The determinations were effected in each case strictly in accordance with the manufacturer's instructions.","1.Standard Experiment PTH IRMA standard of the company Nichols Institute having a nominal concentration of 46.8 pmol/l.","2.Experiment with Plasma Pools Two separate samples of EDTA plasma samples from dialysis patients having increased content of intact PTH were pooled.","Pool PTH intact Elecsys PTH intact Nichols Pool 1 76.7 pmol/l 59.2 pmol/l Pool 2 72.4 pmol/l 54.5 pmol/l The standard sample and the plasma pools where in each case treated with three different concentrations of hPTH(1-37) fragments in five different oxidation and state forms and then the recovered PTH contents determined.","Two treated PTH(1-37) fragment solutions received native hPTH(1-37) from various manufacturers (1: IPF=Institut für Peptidforschung, Braunschweig, 2=Immundiagnostik AG, Bensheim).","In the third fragment solution of hPTH(1-37) the methionine at position 8 was oxidized, in the fourth the methionine at position 18 and in the fifth both methionines at positions 8 and 18 were oxidized.","There were produced from each fragment solution three initial concentrations (1000 nM, 100 nM and 10 nM).","A further dilution of the fragment solutions 1:100 in the standard and in the pool plasma yielded final concentrations of 10000 pMol, 1000 pMol and 100 pMol in the sample to be analyzed.","There were employed in each case three fragment concentrations because the binding capacities of the captor antibodies employed in the commercial tests against the N-terminal PTH fragment were not known.","The aim of the experiment was to completely occupy the captor antibodies with native or oxidized hPTH(1-37), so that they are no longer available for forming the sandwich complex and the measurement signal fell below the respective detection limit.","The comparison of the measurement values from samples having added native hPTH(1-37) fragments or added oxidized fragments then provides information on the specificity of the captor antibodies.","The results are summarized in the diagram shown in FIGS.","1 to 4.For calculating the blockage of the N-terminal antibodies, the measured concentrations of the standard and of the pool plasmas were treated with the native PTH preparation hPTH(1-37) from IPF were determined to be 100%, and all further values placed in relationship thereto.","Discussion of the Results with the Elecsys™2010 PTH Test See diagrams of FIGS.","1 and 2.On the basis of the minimal percentage deviations of the measurement values of all PTH fragments, oxidized at different positions, it can be assumed that the Roche Elecsys™2010 test recognizes the oxidized PTH only with slight affinity and mainly reacts with native and thus biologically active parathyroid hormone.","The absence of signal reduction even with the very high fragment concentration of 10 000 pM speaks for an extremely high binding capacity of the captor antibody.","The native measured samples lay in absolute concentration a maximum of 10% higher than the corresponding treated samples.","With the Elecsys™2010 PTH test it however remains disadvantageous that directly produced active N-terminal PTH fragments are not detected, since the tracer antibody binds in the C-terminal section of the PTH; on the other hand, however, biologically inactive PTH(4-37)-fragments, which are lacking the N-terminal epitope with the amino acids 1 to 3, are partially detected.","Discussion of the Results with the Nichols IRMA PTH Test See the diagrams of FIGS.","3 and 4.Differently than with Elecsys™2010 PTH test the spiking experiments with the IRMA intact PTH test produced clear differences in reaction behaviour depending upon the addition of the various fragments.","The standard employed also behaved differently to the pool plasmas, which may be due to the different matrix (protein content etc.).","The visible signal reduction with a concentration of 1000 pM hPTH(1-37) to 50% of the native measured sample shows the lower binding capacity of the captor antibody against the N-terminal PTH fragment.","The non-oxidized hPTH(1-37) fragments from IPF and Immundiagnostik behave differently, whereby the difference is particularly clear in the case of the pool plasmas.","Also the measurement signals fell clearly as soon as oxidized hPTH(1-37) fragments were added.","On the basis of the results it can be assumed that the polyclonal antibody material employed in the test kit of the Nichols Company contains various antibody populations which react both with native and also with oxidized PTH.","The different binding behaviour of the various oxidized hPTH(1-37) fragments is interesting.","It is to be checked whether the various oxidized molecules (at position 8 or 18 or both positions) have different biological effects.","With the experiment it could be shown that with the polyclonal antibody material (Nichols IRMA), in contrast to the monclonal antibodies (Elecsys 2010), along with native PTH also oxidized inactive PTH forms were detected.","The oxidized PTH has for the target cells for calcium homeostasis (kidney, bone) a restricted to no biological activity.","Example 3 Oxidation and Reduction of Plasma Samples of Uraemic Patients In dialysis, dialysis patients are exposed to an increased oxidation stress.","It was thus investigated whether an oxidation of the PTH occurs in vivo—with corresponding consequences for its biological activity.","It was further investigated whether and to what extent a direct oxidation or a reduction or reverse reduction of the PTH molecules in the samples is possible.","30 plasma samples of dialysis patients and ESRD patients (ESRD=end stage renal disease) were selected at random and oxidized with hydrogen peroxide (30%; end concentration: 1.5 Mol/L) or reduced with sodium boron hydride (NaBH4, aqueous solution of 100 nMol/L).","All samples, treated and untreated, where measured at the same time with the Roche Elecsys™2010 and the Nichols IRMA test.","The oxidation was effected with 30% hydrogen peroxide solution.","The hydrogen peroxide concentration in the incubation preparation was at 17% of the 30% solution.","All samples were incubated for 45 minutes at 37° C., then also cooled down to −70° C., lyophilized and stored at −20° C. until the quantitative PTH determination.","Of 20 samples an aliquot of each in the ratio 1:1 (v/v) was treated and reduced with a 100 nMol NaBH4 solution.","Further, of 10 oxidized samples in each case an aliquot was reverse reduced by the addition of a 100 nMol NaBH4 solution in the ratio 1:1.The samples were incubated for 45 minutes at room temperature and the reaction stopped on dry ice.","After the lyophilization the samples were deep frozen at −20° C. until determination.","The diagrams of FIGS.","5 to 11 show the results.","See FIGS.","5 and 6.The measurement values obtained with the Roche Elecsys™2010 test for native PTH lay, after the oxidation, on average at 29% of the initial value.","Here, the individual samples behaved very differently.","The maximum reduction of the measurement signals was 10% and the minimal reduction of the measurement signal was 53% of the initial value.","The result show that the captor antibody used in the Roche Elecsys™2010 PTH test mainly recognizes the native, reduced PTH form.","See FIGS.","7 and 8.The oxidation has clearly lesser influence on the measurement values in the Nichols IRMA.","On average, the concentration reduced to 76% of the initial value with a maximum reduction to 33% in a minimal reduction 98%.","In order to exclude a partial destruction of the PTH molecule as a result of the pre-treatment of the samples with H2O2, the oxidized samples were reverse reduced again with NaBH4.Upon measurements of the reverse reduced samples with the Elecsys™2010 PTH test only ca.","50% of the initial value could be attained.","Since the detection system is however based on an electrochemical luminescence reaction, the reduction medium probably disrupted determination.","In further experiments a disruption of the measurement signal in the Elecsys system was found also with other antioxidants.","Probably, however, also a proportion of the methionine was oxidized to sulfone, which cannot be reverse reduced.","FIGS.","9 and 10 show the results in the parallel experiments with the Nichols IRMA.","The results show that the oxidation or reduction of the peptide does not influence the immunological activity.","After oxidation and corresponding reverse reduction the native initial concentrations were largely attained.","The slight differences in the recovery after the reverse reduction are probably due to different (oxidation) initial conditions of the peptide.","FIG.","11 shows the results for the sole reduction of native PTH.","The sole reduction leads to a slightly increased recovery (plus 4%) of the native initial value.","The polyclonal antibodies employed in the Nichols test, with their higher affinity for the oxidized form of the peptide, probably prevent a detectable accented manifestation of different PTH concentrations before and after reduction of the peptide.","The data shows clearly the influence of an oxidation on the concentration of intact PTH measured with various tests.","Native and oxidized PTH show significant differences in the biological activity on various target organs.","In particular dialysis patients are clearly exposed to a high oxidative stress and the suspicion lies to hand that free reactive oxygen species significantly change the parathyroid hormone, through oxidation of methionine in its structure and thus significantly changes the biological activity.","Thus, dialysis patients often suffer from hyperhomocystinemea, a known indicator for oxidation stress and a disrupted homocystine metabolism (Durand P. et al.","Laboratory Investigation 2001, 81(5), 645-672).","In the parathyroid hormone the methionines at positions 8 and 18 are above all susceptible to oxidation.","The oxidation of the methionines at positions 8 and 18 to methionyl sulfoxide is probably biologically reversible due to the methionyl sulfoxide reductase occurring in the tissues (Vogt W., Free Radical Biology & Medicine, 1995, 18(1), 93-105).","It leads certainly to a loss of physiological effect (Galceran et al, Endocrinology, 1984, 115(6), 2375-2378; Horiuchi N. J.","Bone Miner.","Res., 1988 3(3), 353-358), Zull J E et al., J. Biol.","Chem., 1990 265(10), 56715676; O'Riordan J L H et al., J.","Endocr., 1974, 63, 117-124).","Thus, the tests available at the present time cannot differentiate biologically active PTH from inactive PTH.","Example 4 Selective Blocking of the N-Terminal Binding Site of Oxidized PTH Through the Addition of Monoclonal Antibodies Against Oxidized PTH There was employed a monoclonal antibody against oxidized PTH in accordance with Logue F C et al.","(Annu.","Clin.","Biochem.","1991, 28, 160-166; J. Immunol.","Methods, 1991, 137, 156-166).","Briefly, this was obtained in that synthetic hPTH(1-34) was oxidized by the addition of hydrogen peroxide (30%) and then DA rats were immunized with this.","There may also be effected an immunization with synthetic PTH fragments (1-37) which contain a methionyl sulfoxide at positions 8 or 18.The fusion of the rat spleen cells with the mouse myeloma line X63Ag8.653 then delivers various monoclonal antibodies which were tested in an immunoassay for their characteristics for binding and masking oxidized PTH.","FIGS.","12 and 13 show elimination experiments with a monoclonal antibody mAK-hPTH(1-37)-metox-8/18 (3B3), which recognizes an epitope having PTH methionyl sulfoxide.","For comparison purposes, this antibody was employed in a conventional PTH IRMA of the Nichols Institute and in the Elecsys™200 PTH of the company Roche.","For this purpose there were produced 10 EDTA plasma pools with mean PTH values in the high concentration region (mean value±SD: NID 46±17 pMol/L; Roche 41±14 pMol/L) and for blocking the N-terminal binding site were treated with the above-described monoclonal antibody against oxidized PTH.","For this purpose, the samples were so mixed with a solution of the monoclonal antibody that the final concentration in the sample with regard to the monoclonal antibody was at 200 ng/ml or 20 ng/ml.","The sample was preincubated for one hour at room temperature with the masking antibody against oxidized PTH and then subjected to determination in conventional manner with the respective test kit from Roche or Nichols.","The results are illustrated in FIGS.","12 and 13.Here, NID and R stand for the Nichols or Roche test and “neat” for untreated samples, 200 for the addition of 200 ng/ml mAK-hPTH(1-37)-metox-8/18 (3B3) masking antibody, 20 for the addition of 20 ng/ml masking antibody, Rox or NIDox for a hydrogen peroxide oxidized sample according to Example 3, and 200 or 20 for the indicated dose of masking antibody.","The results show that the PTH assay of the Nichols Institute cannot distinguish between oxidized inactive PTH and native active PTH, but that through the addition of masking antibodies against oxidized PTH—even with a polyclonal goat antibody against hPTH(1-34)—an antibody binding in the N-terminal section and an incorrect determination can be suppressed to the widest degree.","The experiments further show that even normal pool plasmas contain ca.","10% oxidized PTH.","The Roche PTH test contains a conformation detection antibody (PTH 55-64) which can apparently distinguish between oxidized and native-active PTH.","However, through the addition of masking antibodies falsely excessive determinations, in particular in the untreated sample, are clearly reduced.","It is to be noted that both comparison tests detect no active PTH fragments of the type hPTH(1-34v37) and thus deliver immanently false PTH contents."]],"string":"[\n [\n \"Method for determining effective parathormone activity in a sample\",\n \"Method for determining the active parathyroid hormone content in a sample, wherein the component of parathyroid hormone polypeptide chains which are oxidized at one or more sites near the PTH receptor binding structure having the amino acids 15 to 22 are specifically excluded from the content determination.\",\n \"Oxidation of the parathyroid hormone occurs in particular in dialysis patients, their blood plasma being exposed to an oxidation stress in the dialysis.\",\n \"The invention is based on the use of antibodies which bind specific conformation epitopes of the oxidised or reduced parathyroid hormone and its fragments.\",\n \"Further, there is provided a test system for the quantitative determination of parathyroid hormone and active fragments hereof in a sample.\"\n ],\n [\n \"1.Method for determining the effective parathyroid hormone content in a sample of a body fluid, characterized in that there are specifically excluded from the content determination intact parathyroid hormone polypeptide chains and N-terminal intact fragments hereof which are oxidized at one or more sites near the PTH receptor binding structure having the amino acids 15 to 22.2.Method according to claim 1, wherein the methionine at position 8 or 18 of the parathyroid hormone polypeptide chain, or at both positions, is oxidized.\",\n \"3.Method according to claim 1 or 2, wherein antibodies are employed which bind to a PTH epitope which is formed of at least one oxidized methionine group at position 8 and/or 18.4.Method according to any of claims 1 to 3, wherein antibodies are employed which bind specific parathyroid hormone polypeptide chains or fragments hereof which are oxidized at methionine 8 and/or 18.5.Method according to any of claims 1 to 4, wherein antibodies are employed which bind specific conformation epitopes of the oxidized or native, reduced parathyroid hormone.\",\n \"6.Method according to any of claims 3 to 5, wherein the antibody employed is purified or selected by means of a binding to parathyroid hormone which is oxidized at position 8 and/or 18.7.Method according to any of claims 1 to 6, wherein intact parathyroid hormone and fragments hereof are oxidized and/or reduced and then the total content of intact parathyroid hormone and its active fragments is determined.\",\n \"8.Method according to claim 7, whereby the total content of oxidized and reduced intact parathyroid hormone and its fragments is determined.\",\n \"9.Method according to any preceding claim, wherein the contents of oxidized intact parathyroid hormone and fragments hereof are separately determined.\",\n \"10.Method according to any preceding claim, whereby antibodies according to claim 3 or 4 are put to use for masking of oxidized parathyroid hormone and its fragments, so that exclusively reduced, biologically active parathyroid hormone and its fragments are determined.\",\n \"11.Method according to any preceding, including the following steps: treating the sample with an antibody which recognizes an epitope which is formed from the ends of the N-terminal amino acids 1 to 3 of the parathyroid hormone; treating the sample with an antibody which recognizes an epitope in the region of the receptor binding site between amino acids 15 and 22 of human parathyroid hormone, and determination of the molecules which are recognized by the two antibodies.\",\n \"12.Method according to claim 11, wherein the binding of the two antibodies is effected in the presence of 0.05 to 0.1 weight % of a mild detergent such as Tween™ or Triton™X-100.13.Method according to claim 11 or 12, wherein the sample is treated with antibodies against oxidized parathyroid hormone in accordance with claim 3 or 4, so that antibodies which bind in the region of the receptor binding site of the parathyroid hormone cannot bind to the oxidized parathyroid hormone chain.\",\n \"14.Method according to any preceding claim, wherein the antibodies for masking of oxidized parathyroid hormone and fragments hereof are of a type different from the antibodies for building the sandwich complex.\",\n \"15.Method according to any preceding claim, wherein at least one of the antibodies carries a marking selected from a fluorescing or chemiluminescing group or an enzyme for catalysis of a detection reaction.\",\n \"16.Method according to any preceding claim, wherein one of the antibodies is a monoclonal antibody marked with a ruthenium complex and recognizes an epitope in the region of the amino acids 15 to 22 or 26 to 32.17.Method according to any preceding claim, whereby one of the antibodies is biotinylated.\",\n \"18.Method according to any preceding claim, whereby antibodies are employed which bind to the parathyroid hormone polypeptide chain with amino acids 4 to 14 or 15 to 35v39.19.Test system for the quantative determination of parathyroid hormone and fragments-hereof in a sample, characterized by antibodies which bind to or near to the PTH receptor binding structure of the parathyroid hormone, and by antibodies which bind the epitopes of the parathyroid hormone having oxidized methionine 8 and/or 18.20.Test system according to claim 19, wherein the polyclonal antibody against the N-terminal fragment 1 to 37 of the parathyroid hormone is purified by means of affinity chromatography of antibodies which bind to methionine 8 and/or 18 of the oxidized parathyroid hormone fragment.\",\n \"21.Test system according to claim 19 or 20, wherein there are determined intact parathyroid hormone and N-terminal fragments hereof having the amino acids 1 to 3 in the reduced structure.\",\n \"22.Diagnosis of oxidation stress in dialysis patients, characterized by the employment of an in vitro test for the quantitative determination of parathyroid hormone and its active fragments in accordance with any preceding claim.\"\n ],\n [\n \"The invention relates to an immunological method for determining the effective concentration of parathyroid hormone contained in a sample.\",\n \"The invention relates in particular to a test kit for the determination of the content of active parathyroid hormone and its biologically active fragments for diagnosis, etiology and treatment of disturbances of the calcium metabolism, osteopathies and hyper- or hypo-parathyroidism.\",\n \"The parathyroid hormone (PTH) is formed in the parathyroid gland (Glandulae parathyroideae) and secreted into the blood circulation.\",\n \"In the intact form it consists of a single polypeptide chain having 84 amino acids and has a molecular weight of ca.\",\n \"9500 Dalton (see SWISS-PROT: P01270, PTHY-HUMAN).\",\n \"Together with Vitamin-D and calcitonin it brings about the mobilization of calcium and phosphate out of the bone skeleton and increases the uptake of calcium in the intestines and the excretion of phosphate via the kidneys.\",\n \"The effective PTH activity in plasma or serum is thus an important diagnostic parameter and necessary for determining 1) presence and degree of hyper- or hypo- parathyroidism, 2) quantification of osteoblast activity, 3) quantification of osteoclast activity 4) monitoring of treatment with Vitamin-D and active Vitamin-D metabolites, 5) estimation of presence of aluminium, 6) estimation of a possible oestrogen deficiency in post-menopausal dialysis patients, 6) the necessary steroid or cyclosporin dosage after kidney transplantations, 7) the need for treatment and the prevention of pathological bone marrow changes, uraemic conditions and chronic kidney failure.\",\n \"In the state of the art the determination of PTH is effected by means of the detection of two separated epitopes on the PTH polypeptide chain.\",\n \"With the introduction of specific highly sensitive immunoassays for determining the quantity of intact PTH chains in human serum there appeared to be answered all diagnostic questions in the field of calcium homeostasis, bone transformation and the responsible controlling organs such as the parathyroid gland.\",\n \"However, the values so determined often do not correspond to the actually present PTH activity in the plasma or serum sample, because they often provide PTH contents which are not reconcilable with the symptoms of the patients.\",\n \"In the case of patients with uraemic conditions with normal bone transformation, the levels of intact PTH in serum lie mostly higher by the factor 2.5 than with patients with healthy kidneys (pathological limit in the case of patients with healthy kidneys: 65 μg PTH/L; for patients having uraemic conditions: 165 μg PTH/L serum) Further, uraemic patients with comparably high PTH values often manifest significant differences in bone transformation (Slatopolsky E et al.\",\n \"(2000), Kidney Int., 58, 753-761).\",\n \"Thus these patients often have in the serum eight to ten times increased PTH concentrations, but low normal values for bone specific alkaline phosphatase (ostase).\",\n \"They are thus free from symptoms of an excessive PTH activity.\",\n \"As possible causes there have been discussed either systematic errors in the determinations or a PTH resistance of the osteoblasts, for example due to genetically reduced expression of PTH receptor.\",\n \"Further, the determination of the effective PTH content in serum or plasma is made more difficult in that the hormone is broken down within minutes, in the circulation or the liver, into active and inactive fragments.\",\n \"Some of these fragments have an activity comparable with intact PTH (see EP-A 0 349 545; Schmidt-Gayk et al.\",\n \"(1999) Osteologie forum, 5, 48-58), on the other hand others work antagonistically.\",\n \"Thus, one knows that the fragments hPTH (3-34) and hPTH (7-34) can inhibit the effects of PTH (Suva et al.\",\n \"(1987) Science, 237, 893ff; EP 0 451 867).\",\n \"Moreover, “non-(1-84)” large PTH fragments have been suspected to lie behind the erroneous determinations (LePage R. et al.\",\n \"(1998) Clin.\",\n \"Chem., 44, 805-809).\",\n \"The designation “Large PTH fragment” is used generally for fragments which contain at least the polypeptide chain with the amino acids 39-84 and epitopes of the N-terminal fragments hPTH (1-33), hPTH (1-34), hPTH (1-37) or hPTH (1-38) but from which the N-terminal amino acids 1 to 3 are missing.\",\n \"WO 96/10041 teaches that after loss of the N-terminal amino acids serine and valine the PTH fragments are no longer active.\",\n \"The characterising of various immunological tests with synthetic fragments of the type hPTH (7-84) showed that these as a rule also determine the inactive large PTH (7-84) fragment (John M R et al.\",\n \"(1999), J. Clin.\",\n \"Endocrinol.\",\n \"Metab., 84.4287-4290; Gao P et al.\",\n \"2000, Poster M455, ASBMR 22nd Annual Meeting; Roth H J et al.\",\n \"(2000), Poster P1288; 11th International Congress of Endocrinology, Sydney).\",\n \"The co-determination of inactive large PTH fragments was thus made responsible for the discrepancy between measured PTH concentration and clinical findings.\",\n \"It was suspected that intact PTH molecules compete with the large PTH fragments for the binding sites on the PTH receptor and that depending upon the concentration of CAP (Cyclase activating PTH—biologically active) or CIP (Cyclase Inhibiting PTH—biologically inactive, receptor blocking) different cell activations occurred.\",\n \"It is object of the invention to make available a method for the quantative determination of the effective PTH content of a sample.\",\n \"This object is achieved by means of the method according to claim 1.Further advantageous embodiments of the method are indicated in the subclaims.\",\n \"The method in accordance with the invention for quantitative determination of the PTH content in a sample is characterized in that intact PTH polypeptide chains and N-terminal intact fragments hereof having the PTH receptor binding structure (15 to 22), which are oxidised at one or more sites neighbouring the PTH receptor binding structure (15 to 22), are excluded from the content determination.\",\n \"In other words there are excluded from the determination of the PTH activity or the effective PTH content all PTH polypeptide chains and fragments hereof which are oxidised at methionine-8 or -18 or at both positions.\",\n \"This is effected through the use of antibodies which bind to a PTH epitope which is formed from at least one oxidised methionine group at position 8 and/or 18, or of antibodies which bind specifically PTH polypeptide chains or fragments hereof which are oxidised at methionine 8 and/or 18, or antibodies which can distinguish between specific conformation epitopes of the oxidised and of the “native” non-oxidised parathyroid hormone.\",\n \"It is this ensured that there are excluded from the determination: inactive large PTH fragments of the types hPTH (7-84) or hPTH (3-84), oxidised intact hPTH (1-84) and oxidised N-terminal intact oxidised hPTH (1-34) fragments of the type ox-hPTH (1-34) or ox-hPTH (1-37), which contain the receptor binding structure PTH (15-22).\",\n \"In accordance with the invention, the antibodies are obtainable via a purification or selection on parathyroid hormone which is oxidised at position 8 and/or 18.The antibodies can be obtained through immunization with synthetic or specifically oxidised PTH fragments (Logue F C et al.\",\n \"(1991), Ann.\",\n \"Clin.\",\n \"Biochem., 28 160-166; Journal of Immunological Methods, 39, 159).\",\n \"For example, synthetic PTH (1-34) can be oxidised at the methionines 8 and 18 through the addition of H2O2 and put to use for the immunization.\",\n \"Particularly preferred are monoclonal antibodies against methionine-8, which apparently is oxidized first.\",\n \"In an embodiment of the method in accordance with the invention, the parathyroid hormone and its potentially active fragments are chemically post-oxidized and/or post-reduced and the content of intact parathyroid hormone and its active fragments is determined by a double determination (before and after).\",\n \"In a further embodiment, there is separately and additionally determined the content of oxidised intact parathyroid hormone and fragments hereof in the sample and calculated via the simultaneous determination of total PTH and the content of non-oxidized PTH and its biologically active fragments.\",\n \"In a particularly preferred embodiment of the method there are added antibodies against parathyroid hormone oxidized at position 8 and/or 18, for masking the oxidized parathyroid hormone, so that exclusively non-oxidized or biologically active parathyroid hormone and its fragments are bound by primary and secondary antibodies.\",\n \"The method in accordance with the invention then includes the following steps: treating the sample with an antibody which recognizes an epitope in which parathyroid hormone oxidized at position 8 and/or 18 is involved; treatment of the sample with an antibody which recognizes an epitope which is formed from the N-terminal amino acids 1 to 3 of the parathyroid hormone; treatment of the sample with an antibody which recognizes an epitope in the region of the receptor binding structure (amino acids 15 to 22) of human parathyroid hormone and which cannot bond when the antibody has already bonded in the region of the oxidized methionine 8 and/or 18, and determination of the number of the molecules which are recognized by the two antibodies.\",\n \"Preferably, here the binding of the two antibodies takes place in the presence of the 0.05 to 0.1 weight percent of a mild detergent such as Tween™20 or Triton™X-100.In accordance with the invention, in a particularly preferred embodiment, the sample is thus first treated with antibodies against oxidized parathyroid hormone, so that antibodies which bind in the region of receptor binding site of the parathyroid hormone cannot bind to the oxidized PTH polypeptide chain.\",\n \"The person skilled in the art will recognize that the antibody for masking of oxidized parathyroid hormone and fragments hereof should better, be of a different type than the antibody for forming or detecting the sandwich complex.\",\n \"The antibodies employed as captors and tracers, may, as the skilled person knows, also be functionally exchanged.\",\n \"In a further embodiment at least one of the antibodies carries a marking, selected from a fluorescing or chemiluminescing group or an enzyme for catalysing a detection reaction.\",\n \"Preferably one of the antibodies is a monoclonal antibody marked with a ruthenium complex, which recognizes an epitope in the region of the amino acids 15 to 22 or 26 to 32.One of the antibodies may also be biotinylated and bonded via streptavidin to a solid phase.\",\n \"In the method there can further be employed antibodies which bind to the PTH polypeptide chain with amino acids 4 to 14 or 15 to 35v39.An antibody which recognizes an epitope in the range of amino acids 15 to 22 is particularly preferred since on the one hand this epitope lies in the range of the receptor binding structure, on the other hand such an antibody can distinguish, due to the methionine 18, between oxidized and native hPTH.\",\n \"The test system in accordance with the invention for the quantitive determination of parathyroid hormone and fragments hereof in a sample includes in one embodiment antibodies (captor antibodies) bound to a phase, which antibodies bind in or near the PTH receptor binding structure of the parathyroid hormone and—for masking—free (unbound) antibodies, which bind the epitopes of the parathyroid hormone having oxidized methionine 8 and/or 18.The antibodies can be purified through affinity chromatography, for example via their binding to methionine 8 and/or 18 of the oxidized parathyroid hormone.\",\n \"As tracer antibodies there are employed antibodies against the N-terminal amino acids 1 to 3 of the parathyroid hormone (in the reduced structure).\",\n \"Captor and tracer antibodies may also be exchanged.\",\n \"The invention further relates to the diagnosis of oxidation stress in dialysis patients, whereby an in vitro test for the quantitative determination of parathyroid hormone and it active fragments in accordance with the invention is put to use.\",\n \"Even though one knew from experiments with synthetic PTH fragments that oxidized parathyroid hormone or oxidized parathyroid hormone fragments are biologically inactive, one did not imagine that this oxidation could assume physiologically relevant levels and prevent the determination of the true physiological PTH activity in a sample.\",\n \"Thus in some conventional test methods there were added reduction means for stabilizing the antibodies in the sample buffer, in others a subsequent partial oxidation of the PTH molecule was tolerated.\",\n \"The invention also relates in an embodiment to a method for a diagnosis and determination of the extent of a hypo- or hyper-parathyroidism, the etiology of calcium metabolism disturbances, osteopathies, kidney failures and conditions which arise from a disturbed homeostasis of the calcium and phosphate contents of the blood.\",\n \"Further, in accordance with the invention the direct production of N-terminal PTH fragments, so far as this arises, is also detected.\",\n \"This means that the PTH diagnosis in accordance with the invention detects along with intact active parathyroid hormone also directly produced, biologically active PTH fragments.\",\n \"Oxygen is essential for aerobic life but at the same time is the most important precursor molecule of free reactive oxygen species (ROS) which through their reaction with proteins, lipids and nucleic acids damage the cells and through the oxidation of reactive amino acids can alter proteins in their structure.\",\n \"Many publications have been concerned with the effect of oxidation stress and the influence of “advanced glycation end products” on the pathogenesis of complications in the case of chronically kidney insufficient patients (Martin-Mateo M C et al.\",\n \"(1999), Ren.\",\n \"Fail., 21, 155-167; Hasselwander O et al.\",\n \"(1998), Free Radic.\",\n \"Res., 29, 1-11; Zoccali C et al.\",\n \"(2000), Nephrol.\",\n \"Dial.\",\n \"Transplant., 15, Suppl.2; Canaud B et al.\",\n \"(1999), Blood Purif., 17, 99-106).\",\n \"Dialysis patients are in comparison to normal persons much more strongly affected by oxidative stress.\",\n \"Various working groups have investigated oxidized parathyroid hormone and its biological activity (Alexiewicz L M et al.\",\n \"(1990), J.\",\n \"Am.\",\n \"Soc.\",\n \"Nephrol., 1, 236-244; Zull J E et al.\",\n \"(1990), J. Biol.\",\n \"Chem., 265, 5671-5676; Pitts T O et al.\",\n \"(1988), Miner.\",\n \"Electrolyte Metab., 15, 267-275; Horiuchi N. (1988), J.\",\n \"Bone Miner.\",\n \"Res., 3, 353-358; Frelinger A L et al.\",\n \"(1986) Arch.\",\n \"Biochem.\",\n \"Biophys., 244, 641-649; Galceran T et al.\",\n \"(1984) Endocrinology 115, 2375-2378; Frelinger A L et al.\",\n \"(1984), J. Biol.\",\n \"Chem., 259, 5507-5513; O'Riordan J L H et al.\",\n \"(1974), J.\",\n \"Endocr., 63, 117-124; Logue F C et al.\",\n \"(1991), Ann.\",\n \"Clin.\",\n \"Biochem., 28, 160-166; Logue F C (1991b), J. Immun.\",\n \"Meth., 39, 159).\",\n \"Oxidation stress in dialysis patients and its consequence for PTH diagnosis have, however, not so far been investigated and recognized.\",\n \"It has now been found that the conventional methods of determination, in each case depending on the test kit and antibodies, detect both bioactive (native or reduced) intact PTH and if applicable its fragments and also oxidized PTH and if applicable its fragments, and thus falsely determine the effective level of parathyroid hormone.\",\n \"Further, it has been found that the postulated large PTH fragments, insofar as they arise at all, have no biological significance: rather that the discrepancy between measured PTH content and medical determination arises at least partly from the oxidation stress to which the dialysis patient is subject.\",\n \"The effective PTH content in a plasma or serum sample is namely dependent upon the oxidation stress and the oxidation stress is at the same time a cause of pathological conditions.\",\n \"The invention relates further to a method in which a first captor antibody is bonded to a solid phase.\",\n \"The second tracer antibody may carry a marker, e.g.\",\n \"a fluorescent or chemiluminescent group, or may be conjugated with an enzyme such as alkaline phosphatase or peroxidase.\",\n \"The method is effected in accordance with the invention in a per se known immunoassay, preferably in an ELISA, ECLIA, IEMA, IRMA, ILMA or LIA.\",\n \"The binding of the two antibodies to the PTH is preferably effected in the presence of 0.05 to 0.1 weight percent of a mild detergent such as Tween™20 or Triton™X-100 so that a folding of the N-terminal epitope PTH (1-3) to the PTH receptor binding structure is prevented, or in order to keep the N-terminal epitope PTH (1-3) accessible for binding with the antibody.\",\n \"Since for the biological activity and the folding of the PTH both the receptor binding structure and also the amino-terminal epitope PTH (1-3) are necessary, these two structures probably interact with one another.\",\n \"Through the presence of a mild detergent, in the above-mentioned embodiment with antibodies against hPTH (1-3), the sensitivity and accuracy of the assay can be significantly improved.\",\n \"In an embodiment of the method a known aliquot of the sample is further treated with antibodies which bind to the PTH sequences between amino acids 4 to 14 and/or 15 to 37.The number of molecules in a sample which are bound by antibodies against the epitope PTH (1-3) and the receptor binding structure of the PTH, and the number of molecules which are bound by the antibodies against the PTH regions 4 to 14 and 15 to 37, are then placed in relationship one to another and the biologically effective PTH activity determined.\",\n \"The invention further relates to a diagnosis system for the determination of the PTH activity in a sample, which system is characterized by antibodies which recognize specifically the N-terminal epitope with the amino acids 1 to 3 and antibodies which bind to the region of the PTH receptor binding structure.\",\n \"In a further embodiment the diagnosis system further has antibodies which bind in the region between amino acids 4 to 14.In a third embodiment the system includes antibodies which bind in the section between amino acids 24 and 37 of the parathyroid hormone or on the middle (53 to 86) or the C-terminal (53 to 84) section of the PTH polypeptide chain.\",\n \"In contrast to earlier immunoassays, the method in accordance with the invention takes into account the PTH activity also of the non-intact PTH fragments and that apparently intact PTH is biologically inactive when the last or last two amino terminal amino acids of the parathyroid hormone are missing or when one or both methionines at position 8 and 18 of the N-terminal fragments are oxidized.\",\n \"PTH fragments which have neither a receptor binding structure with the amino acids 15 to 22 nor an intact N-terminal epitope PTH (1-3) have no agonistic or antagonistic activity.\",\n \"Conventional immunological tests for the quantative determination of intact parathyroid hormone or hPTH (1-37) yield false activity values if a physiological activity is concluded upon only from the presence of two epitopes.\",\n \"Conventional methods do not take into account that the correct folding or the binding to the PTH receptor is dependent upon an intact amino-terminus with the amino acids serine and valine and that the methionine groups at position 8 and 18 near the PTH receptor binding structure must not be oxidized.\",\n \"The invention rests in particular on the discovery that the oxidation of PTH at the above-mentioned positions, in particular in the case of dialysis patients, can be considerable and of diagnostic significance.\",\n \"The fragments hPTH (7-84), hPTH (3-84) or hPTH (4-37), so far as they are present, determined in conventional tests as active PTH have no or no measurable antagonistic activity.\",\n \"Test systems on the basis of antibodies against the middle section (53-68) or the C-terminal (53-84) section overlook the fact that some patients directly produce biologically active PTH fragments of the type hPTH (1-37), hPTH(1-32˜36) or hPTH(1-38).\",\n \"Immunological methods on the basis of antibodies against hPTH(1-3) or hPTH(1-6), or against peptides from the hPTH(1-37) fragments, also reflect false activities since there are determined not only the biologically active (reduced) structure units.\",\n \"With polyclonal antibodies or antisera against the N-terminal peptide having the amino acids 1 to 5 or 6 in part even antagonistically effective fragments are recognized as active PTH fragments.\",\n \"The method in accordance with the invention thus makes available to the doctor a PTH activity value which indicates the physiological effects of the native (reduced) parathyroid hormone and its active fragments.\",\n \"It recognizes also patients hyperparathyroidism who have normal or reduced contents of intact hPTH(1-84) in the serum and directly secrete PTH fragments such as hPTH(1-37) and hPTH(1-38) into the bloodstream.\",\n \"The method in accordance with the invention for determining the activity of PTH and its fragments in a sample can be realized as EIA, ELISA, RIA, IRMA, LIA or ICLIA, ILMA, FIA or IFMA, as a manual test system or preferably in a version adapted to automatic systems, in liquid phase or solid phase techniques.\",\n \"Further advantages, features and embodiments of the invention are provided through the following examples and the accompanying drawings.\",\n \"There is shown: FIG.\",\n \"1 a diagram with results (test system: Elecsys™2010 of Roche Diagnostics GmbH, Mannheim) of spiking experiments on PTH standard; FIG.\",\n \"2 a diagram with results (Elecsys™2010 of Roche Diagnostics) of spiking experiments on pool plasmas; FIG.\",\n \"3 a diagram with results (test system: PTH-IRMA of the Nichols Institute, U.S.A.) of spiking experiments on PTH standard; FIG.\",\n \"4 a diagram with results (PTH-IRMA of the Nichols Institute) of spiking experiments on pool plasmas; FIG.\",\n \"5 a diagram with measured values (Roche Elecsys™2010 of the plasma samples after oxidation with H2O2; FIG.\",\n \"6 a diagram with percentage measurement values (Roche Elecsys™2010) of the plasma samples after oxidation with H2O2; FIG.\",\n \"7 a diagram with measurement values (PTH-IRMA of the Nichols Institute) of the plasma samples after oxidation with H2O2; FIG.\",\n \"8 a diagram with percentage measurement values (PTH-IRMA of the Nichols Institute) of the plasma samples after oxidation with H2O2; FIG.\",\n \"9 a diagram having measurement values (PTH-IRMA of the Nichols Institute) after oxidation and reverse reduction of the samples; FIG.\",\n \"10 a diagram with percentage measurement values (PTH-IRMA of the Nichols Institute) after oxidation and reverse reduction of the samples; FIG.\",\n \"11 a diagram with percentage measurement values (PTH-IRMA of the Nichols Institute) after oxidation and direct reduction of the samples; FIG.\",\n \"12 a diagram with percentage measurement values (PTH-IRMA of the Nichols Institute) of a plasma sample with and without oxidization and after addition of monoclonal antibody against oxidized parathyroid hormone; FIG.\",\n \"13 a diagram with percentage measurement values from a plasma sample (Roche Elecsys™) with and without oxidization and after addition of monoclonal antibody against oxidized parathyroid hormone.\",\n \"EXAMPLES Example 1 Biological Effect of Large PTH Fragments It was investigated whether the large PTH fragments, postulated in the state of the art, occur in human serum and have an antagonistic effect on the PTH receptor.\",\n \"The immunoassays employed were based upon antibodies which do not cross-react with synthetic hPTH(7-84) (Bachem AG), so that the measurement values should indicate the true concentration of intact active hPTH in human serum.\",\n \"These values were compared with parameters for bone transformation.\",\n \"The production of the large PTH fragments is, in accordance with the state of the art, regulated via a feedback mechanism and in the case of increased or reduced need the stimulation or suppression of the parathyroid gland leads to an adequate expression.\",\n \"At the present time there are known two test systems for the quantative determination of PTH which have no clinically relevant cross-reactivity with the synthetic large PTH fragment hPTH(7-84).\",\n \"These are the Elecsys™2010 PTH immunoassay of Roche Diagnostics GmbH, Mannheim and the CAP-PTH-IRMA of the company Scantibodies.\",\n \"The electrochemiluminescence immunoassay (Elecsys™ of Roche Diagnostics GmbH, Mannheim) is based upon a monoclonal captor antibody against an N-terminal PTH epitope corresponding to the amino acids 26-32 and a ruthenium complex marked tracer antibody against a C-terminal hPTH epitope corresponding to the amino acids 55-64.The employed synthetic large hPTH(7-84) fragment (Bachem AG) is not recognized by the captor antibody of the Elecsys™2010 PTH immunoassay, so that it must recognize a tertiary folding structure or is a so-called conformation antibody.\",\n \"Probably, the large hPTH(7-84) fragment employed (Gao P et al.\",\n \"2000, Poster M455, ASBMR 22nd Annual Meeting, Roth H J et al.\",\n \"(2000), Poster P1288; 11th International Congress of Endocrinology, Sydney), as the present results make evident, was oxidized in the course of synthesis or storage.\",\n \"The captor antibody of the Elecsys™2010 PTH immunoassay probably does not bind only oxidized hPTH(7-84), but does bind reduced PTH(7-84).\",\n \"The CAP-PTH-IRMA of the company Scantibodies employs polyclonal antibodies against the N-terminal hPTH(1-6) which could not bind the large PTH(7-84) fragments.\",\n \"Along with the determination of intact PTH using various immunoassays (Nichols IRMA, Roche Elecsys™2010, DUO-PTH Scantibodies, PTH-Immundiagnostik) there were also determined the bone marker BAP (ostase) as a formation marker and TRAP5b as resorption marker.\",\n \"The results are indicated in the following Table 1.TABLE 1 Correlation of bone growth and reduction markers with PTH measurement values Dependent Independent Correlation variable Y variable X coefficient/n= BAP (Ostase) PTH CAP Scan.\",\n \"0.484 n = 96 BAP (Ostase) PTH Elecsys2010 0.474 n = 96 BAP (Ostase) PTH Nichols 0.485 n = 96 TRAP5b PTH CAP Scan 0.511 n = 95 TRAP5b PTH Elecsys2010 0.510 n = 95 TRAP5b PTH Nichols 0.524 n = 95 PTH CAP Scan PTH Elecsys2010 0.966 n = 103 PTH CAP Scan PTH Nichols 0.979 n = 103 The correlations of the values of the growth and reduction markers and the PTH contents in the serum show no significantly greater correlations for the more specific tests (“no cross reactivity to large PTH fragments”) in comparison to the IRMA of the company Nichols Institute.\",\n \"Amongst one another the correlation of the individual PTH assays is, as has also been shown in other studies, excellent.\",\n \"This shows that none of the known test systems provide satisfactory results.\",\n \"Example 2 Oxidation and Reduction Experiments It was investigated whether and to what extent the Elecsys™2010 PTH test and the Nichols Intact PTH test recognize native or oxidized PTH.\",\n \"The Nichols Intact PTH Assay (Nichols Institute, San Juan Capistrano, Calif., U.S.A) is a double binding immunoradiometry assay (IRMA) with two polyclonal goat antibodies.\",\n \"The captor antibody, immobilized in plastic beads, binds only in the middle or C-terminal section of the hPTH (anti-hPTH(39-89)).\",\n \"The second detection antibody, marked with 125Iodine, recognizes an epitope in the N-terminal section of the PTH (anti-hPTH(1-34)).\",\n \"The sample was incubated at the same time with the antibody coated beads and the 125Iodine marked detection antibody overnight (12±2h) at room temperature.\",\n \"After the incubation, the beads were washed and the radioactivity adhering to the solid phase measured in a gamma counter (LKB1277 Gamma Master from Perkin-Elmer).\",\n \"The dose measurement curve (MultiCalc Software) between bound radioactivity and concentration in the sample was determined strictly in accordance with the manufacturer's instructions via a standard measured at the same time.\",\n \"The interassay variance in the normal and increased region was less than 10% CV.\",\n \"The Roche Elecsys™ PTH assay is an electrochemiluminescence immunoassay (ECLIA) on the fully automatic immunoanalyzer Elecsys R2010.The epitope recognized by the biotinylated captor antibody contains the PTH amino acids 26 to 32 and the monoclonal ruthenium detection antibody recognizes, in accordance with the information from the manufacturer, an epitope in the region of the PTH amino acids 55 to 64.Sample and antibodies were incubated together for 9 minutes at 37° C., Streptavidin coated paramagnetic beads introduced to the incubation mixture, and finally incubated for further 9 minutes.\",\n \"After magnetic separation of the bonded conjugates from the free conjugates on an electrode surface the luminescence was determined and the end PTH concentration determined via a recalibrated master curve.\",\n \"The interassay variance in the lower and upper normal region was less than 7% CV and in the high concentration region 5% CV.\",\n \"For the oxidation and reduction experiments standards and pool plasmas from dialysis patients having hPTH (1-37) fragments of different concentrations and oxidation states were treated.\",\n \"All samples received EDTA as anticoagulant.\",\n \"The measurement of the native, unaltered initial samples (reference parameter) and of the treated pool plasmas was effected at the same time.\",\n \"The determinations were effected in each case strictly in accordance with the manufacturer's instructions.\",\n \"1.Standard Experiment PTH IRMA standard of the company Nichols Institute having a nominal concentration of 46.8 pmol/l.\",\n \"2.Experiment with Plasma Pools Two separate samples of EDTA plasma samples from dialysis patients having increased content of intact PTH were pooled.\",\n \"Pool PTH intact Elecsys PTH intact Nichols Pool 1 76.7 pmol/l 59.2 pmol/l Pool 2 72.4 pmol/l 54.5 pmol/l The standard sample and the plasma pools where in each case treated with three different concentrations of hPTH(1-37) fragments in five different oxidation and state forms and then the recovered PTH contents determined.\",\n \"Two treated PTH(1-37) fragment solutions received native hPTH(1-37) from various manufacturers (1: IPF=Institut für Peptidforschung, Braunschweig, 2=Immundiagnostik AG, Bensheim).\",\n \"In the third fragment solution of hPTH(1-37) the methionine at position 8 was oxidized, in the fourth the methionine at position 18 and in the fifth both methionines at positions 8 and 18 were oxidized.\",\n \"There were produced from each fragment solution three initial concentrations (1000 nM, 100 nM and 10 nM).\",\n \"A further dilution of the fragment solutions 1:100 in the standard and in the pool plasma yielded final concentrations of 10000 pMol, 1000 pMol and 100 pMol in the sample to be analyzed.\",\n \"There were employed in each case three fragment concentrations because the binding capacities of the captor antibodies employed in the commercial tests against the N-terminal PTH fragment were not known.\",\n \"The aim of the experiment was to completely occupy the captor antibodies with native or oxidized hPTH(1-37), so that they are no longer available for forming the sandwich complex and the measurement signal fell below the respective detection limit.\",\n \"The comparison of the measurement values from samples having added native hPTH(1-37) fragments or added oxidized fragments then provides information on the specificity of the captor antibodies.\",\n \"The results are summarized in the diagram shown in FIGS.\",\n \"1 to 4.For calculating the blockage of the N-terminal antibodies, the measured concentrations of the standard and of the pool plasmas were treated with the native PTH preparation hPTH(1-37) from IPF were determined to be 100%, and all further values placed in relationship thereto.\",\n \"Discussion of the Results with the Elecsys™2010 PTH Test See diagrams of FIGS.\",\n \"1 and 2.On the basis of the minimal percentage deviations of the measurement values of all PTH fragments, oxidized at different positions, it can be assumed that the Roche Elecsys™2010 test recognizes the oxidized PTH only with slight affinity and mainly reacts with native and thus biologically active parathyroid hormone.\",\n \"The absence of signal reduction even with the very high fragment concentration of 10 000 pM speaks for an extremely high binding capacity of the captor antibody.\",\n \"The native measured samples lay in absolute concentration a maximum of 10% higher than the corresponding treated samples.\",\n \"With the Elecsys™2010 PTH test it however remains disadvantageous that directly produced active N-terminal PTH fragments are not detected, since the tracer antibody binds in the C-terminal section of the PTH; on the other hand, however, biologically inactive PTH(4-37)-fragments, which are lacking the N-terminal epitope with the amino acids 1 to 3, are partially detected.\",\n \"Discussion of the Results with the Nichols IRMA PTH Test See the diagrams of FIGS.\",\n \"3 and 4.Differently than with Elecsys™2010 PTH test the spiking experiments with the IRMA intact PTH test produced clear differences in reaction behaviour depending upon the addition of the various fragments.\",\n \"The standard employed also behaved differently to the pool plasmas, which may be due to the different matrix (protein content etc.).\",\n \"The visible signal reduction with a concentration of 1000 pM hPTH(1-37) to 50% of the native measured sample shows the lower binding capacity of the captor antibody against the N-terminal PTH fragment.\",\n \"The non-oxidized hPTH(1-37) fragments from IPF and Immundiagnostik behave differently, whereby the difference is particularly clear in the case of the pool plasmas.\",\n \"Also the measurement signals fell clearly as soon as oxidized hPTH(1-37) fragments were added.\",\n \"On the basis of the results it can be assumed that the polyclonal antibody material employed in the test kit of the Nichols Company contains various antibody populations which react both with native and also with oxidized PTH.\",\n \"The different binding behaviour of the various oxidized hPTH(1-37) fragments is interesting.\",\n \"It is to be checked whether the various oxidized molecules (at position 8 or 18 or both positions) have different biological effects.\",\n \"With the experiment it could be shown that with the polyclonal antibody material (Nichols IRMA), in contrast to the monclonal antibodies (Elecsys 2010), along with native PTH also oxidized inactive PTH forms were detected.\",\n \"The oxidized PTH has for the target cells for calcium homeostasis (kidney, bone) a restricted to no biological activity.\",\n \"Example 3 Oxidation and Reduction of Plasma Samples of Uraemic Patients In dialysis, dialysis patients are exposed to an increased oxidation stress.\",\n \"It was thus investigated whether an oxidation of the PTH occurs in vivo—with corresponding consequences for its biological activity.\",\n \"It was further investigated whether and to what extent a direct oxidation or a reduction or reverse reduction of the PTH molecules in the samples is possible.\",\n \"30 plasma samples of dialysis patients and ESRD patients (ESRD=end stage renal disease) were selected at random and oxidized with hydrogen peroxide (30%; end concentration: 1.5 Mol/L) or reduced with sodium boron hydride (NaBH4, aqueous solution of 100 nMol/L).\",\n \"All samples, treated and untreated, where measured at the same time with the Roche Elecsys™2010 and the Nichols IRMA test.\",\n \"The oxidation was effected with 30% hydrogen peroxide solution.\",\n \"The hydrogen peroxide concentration in the incubation preparation was at 17% of the 30% solution.\",\n \"All samples were incubated for 45 minutes at 37° C., then also cooled down to −70° C., lyophilized and stored at −20° C. until the quantitative PTH determination.\",\n \"Of 20 samples an aliquot of each in the ratio 1:1 (v/v) was treated and reduced with a 100 nMol NaBH4 solution.\",\n \"Further, of 10 oxidized samples in each case an aliquot was reverse reduced by the addition of a 100 nMol NaBH4 solution in the ratio 1:1.The samples were incubated for 45 minutes at room temperature and the reaction stopped on dry ice.\",\n \"After the lyophilization the samples were deep frozen at −20° C. until determination.\",\n \"The diagrams of FIGS.\",\n \"5 to 11 show the results.\",\n \"See FIGS.\",\n \"5 and 6.The measurement values obtained with the Roche Elecsys™2010 test for native PTH lay, after the oxidation, on average at 29% of the initial value.\",\n \"Here, the individual samples behaved very differently.\",\n \"The maximum reduction of the measurement signals was 10% and the minimal reduction of the measurement signal was 53% of the initial value.\",\n \"The result show that the captor antibody used in the Roche Elecsys™2010 PTH test mainly recognizes the native, reduced PTH form.\",\n \"See FIGS.\",\n \"7 and 8.The oxidation has clearly lesser influence on the measurement values in the Nichols IRMA.\",\n \"On average, the concentration reduced to 76% of the initial value with a maximum reduction to 33% in a minimal reduction 98%.\",\n \"In order to exclude a partial destruction of the PTH molecule as a result of the pre-treatment of the samples with H2O2, the oxidized samples were reverse reduced again with NaBH4.Upon measurements of the reverse reduced samples with the Elecsys™2010 PTH test only ca.\",\n \"50% of the initial value could be attained.\",\n \"Since the detection system is however based on an electrochemical luminescence reaction, the reduction medium probably disrupted determination.\",\n \"In further experiments a disruption of the measurement signal in the Elecsys system was found also with other antioxidants.\",\n \"Probably, however, also a proportion of the methionine was oxidized to sulfone, which cannot be reverse reduced.\",\n \"FIGS.\",\n \"9 and 10 show the results in the parallel experiments with the Nichols IRMA.\",\n \"The results show that the oxidation or reduction of the peptide does not influence the immunological activity.\",\n \"After oxidation and corresponding reverse reduction the native initial concentrations were largely attained.\",\n \"The slight differences in the recovery after the reverse reduction are probably due to different (oxidation) initial conditions of the peptide.\",\n \"FIG.\",\n \"11 shows the results for the sole reduction of native PTH.\",\n \"The sole reduction leads to a slightly increased recovery (plus 4%) of the native initial value.\",\n \"The polyclonal antibodies employed in the Nichols test, with their higher affinity for the oxidized form of the peptide, probably prevent a detectable accented manifestation of different PTH concentrations before and after reduction of the peptide.\",\n \"The data shows clearly the influence of an oxidation on the concentration of intact PTH measured with various tests.\",\n \"Native and oxidized PTH show significant differences in the biological activity on various target organs.\",\n \"In particular dialysis patients are clearly exposed to a high oxidative stress and the suspicion lies to hand that free reactive oxygen species significantly change the parathyroid hormone, through oxidation of methionine in its structure and thus significantly changes the biological activity.\",\n \"Thus, dialysis patients often suffer from hyperhomocystinemea, a known indicator for oxidation stress and a disrupted homocystine metabolism (Durand P. et al.\",\n \"Laboratory Investigation 2001, 81(5), 645-672).\",\n \"In the parathyroid hormone the methionines at positions 8 and 18 are above all susceptible to oxidation.\",\n \"The oxidation of the methionines at positions 8 and 18 to methionyl sulfoxide is probably biologically reversible due to the methionyl sulfoxide reductase occurring in the tissues (Vogt W., Free Radical Biology & Medicine, 1995, 18(1), 93-105).\",\n \"It leads certainly to a loss of physiological effect (Galceran et al, Endocrinology, 1984, 115(6), 2375-2378; Horiuchi N. J.\",\n \"Bone Miner.\",\n \"Res., 1988 3(3), 353-358), Zull J E et al., J. Biol.\",\n \"Chem., 1990 265(10), 56715676; O'Riordan J L H et al., J.\",\n \"Endocr., 1974, 63, 117-124).\",\n \"Thus, the tests available at the present time cannot differentiate biologically active PTH from inactive PTH.\",\n \"Example 4 Selective Blocking of the N-Terminal Binding Site of Oxidized PTH Through the Addition of Monoclonal Antibodies Against Oxidized PTH There was employed a monoclonal antibody against oxidized PTH in accordance with Logue F C et al.\",\n \"(Annu.\",\n \"Clin.\",\n \"Biochem.\",\n \"1991, 28, 160-166; J. Immunol.\",\n \"Methods, 1991, 137, 156-166).\",\n \"Briefly, this was obtained in that synthetic hPTH(1-34) was oxidized by the addition of hydrogen peroxide (30%) and then DA rats were immunized with this.\",\n \"There may also be effected an immunization with synthetic PTH fragments (1-37) which contain a methionyl sulfoxide at positions 8 or 18.The fusion of the rat spleen cells with the mouse myeloma line X63Ag8.653 then delivers various monoclonal antibodies which were tested in an immunoassay for their characteristics for binding and masking oxidized PTH.\",\n \"FIGS.\",\n \"12 and 13 show elimination experiments with a monoclonal antibody mAK-hPTH(1-37)-metox-8/18 (3B3), which recognizes an epitope having PTH methionyl sulfoxide.\",\n \"For comparison purposes, this antibody was employed in a conventional PTH IRMA of the Nichols Institute and in the Elecsys™200 PTH of the company Roche.\",\n \"For this purpose there were produced 10 EDTA plasma pools with mean PTH values in the high concentration region (mean value±SD: NID 46±17 pMol/L; Roche 41±14 pMol/L) and for blocking the N-terminal binding site were treated with the above-described monoclonal antibody against oxidized PTH.\",\n \"For this purpose, the samples were so mixed with a solution of the monoclonal antibody that the final concentration in the sample with regard to the monoclonal antibody was at 200 ng/ml or 20 ng/ml.\",\n \"The sample was preincubated for one hour at room temperature with the masking antibody against oxidized PTH and then subjected to determination in conventional manner with the respective test kit from Roche or Nichols.\",\n \"The results are illustrated in FIGS.\",\n \"12 and 13.Here, NID and R stand for the Nichols or Roche test and “neat” for untreated samples, 200 for the addition of 200 ng/ml mAK-hPTH(1-37)-metox-8/18 (3B3) masking antibody, 20 for the addition of 20 ng/ml masking antibody, Rox or NIDox for a hydrogen peroxide oxidized sample according to Example 3, and 200 or 20 for the indicated dose of masking antibody.\",\n \"The results show that the PTH assay of the Nichols Institute cannot distinguish between oxidized inactive PTH and native active PTH, but that through the addition of masking antibodies against oxidized PTH—even with a polyclonal goat antibody against hPTH(1-34)—an antibody binding in the N-terminal section and an incorrect determination can be suppressed to the widest degree.\",\n \"The experiments further show that even normal pool plasmas contain ca.\",\n \"10% oxidized PTH.\",\n \"The Roche PTH test contains a conformation detection antibody (PTH 55-64) which can apparently distinguish between oxidized and native-active PTH.\",\n \"However, through the addition of masking antibodies falsely excessive determinations, in particular in the untreated sample, are clearly reduced.\",\n \"It is to be noted that both comparison tests detect no active PTH fragments of the type hPTH(1-34v37) and thus deliver immanently false PTH contents.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473975"}}},{"rowIdx":775,"cells":{"text":{"kind":"list like","value":[["Immunotherapeutic combinations for the treatment of tumours that overexpress gangliosides","The present invention is related to the field of immunology and more specifically with immunotherapeutic combinations used for controlling growth and/or cell proliferation of tumors.","By means of the present invention a therapeutic effect against tumors is observed by the combination of idiotypic vaccines whose principle active is an anti-ganglioside antibody (Ab1), with idiotypic vaccines whose principle active is anti-idiotype antibody (Ab2) obtained against an anti-ganglioside antibody, or vaccines whose principle active is one or more gangliosides.","Combinations of these vaccines are described that cause a synergic therapeutic effect against tumors.","The referred combinations can be applied to patients in different clinical conditions of tumors that express gangliósidos."],["1.An immunotherapeutic combination for the immunotherapy of tumors that over-express gangliosides, comprising at least two of the following components: (A) An idiotypic vaccine comprising a murine anti-ganglioside monoclonal antibody (Ab1); (B) An idiotypic vaccine comprising an specific murine anti-idiotype monoclonal antibody (Ab2), against an anti-ganglioside monoclonal antibody; (C) A ganglioside vaccine.","2.An immunotherapeutic combination according to claim 1, wherein said combination comprises A plus B or A plus C or B plus C. 3.An immunotherapeutic combination according to claim 1, wherein said vaccine A comprises the murine anti-ganglioside MAb P3 (Deposit Number ECACC 94113026).","4.An immunotherapeutic combination according to claim 1, wherein said vaccine B comprises the murine anti-idiotype antibody 1E10 (Deposit Number ECACC 97112901).","5.An immunotherapeutic combination according to claim 1, wherein said vaccine C comprises the NeuGcGM3 ganglioside.","6.Immunotherapeutic combination according to claim 1, wherein said vaccine C comprises the NeuAcGM3 ganglioside.","7.Immunotherapeutic combination according to claims 1, wherein A and B or A and C or B and C are administered simultaneously or alternately.","8.Use of any of the immunotherapeutic combinations of the claim 1 for the treatment of tumors that over-express gangliosides.","9.Use according to claim 8, for treatment of tumor of breast, lung, digestive system, urogenital system, melanomas, sarcomas and those from neuroectodermic tissues.","10.A method for controlling growth and/or cell proliferation of tumors that over-express gangliosides, comprising the administration to a mammal of an immunotherapeutic combination of claim 7.11.Method according to claim 10, wherein said mammals are human."],["<SOH> TECHNICAL FIELD <EOH>This invention relates to human medicine, and especially to therapeutic vaccines that induce an immune response to tumors that over-express gangliosides."],["<SOH> BRIEF DESCRIPTION OF THE FIGURES <EOH>FIG.","1 .","Mice Balb/c euthymic and athymic mice were immunized subcutaneously with 100 μg of the murine MAb P3 in CFA followed by a re-immunization with 50 μg of the MAb emulsified in IFA.","After three days, lymph nodes cells were collected and it was carried out the proliferation assay with different concentrations of the MAbs P3, A3, 1E10 and C5.FIG.","2 .","Mice Balb/c were immunized subcutaneously with 100 pg of the chimeric antibody P3 in CFA followed by a re-immunization with 50 μg of the antibody emulsified in IFA.","After three days, lymph nodes cells were collected and it was carried out the proliferation assay with different concentrations using murine and chimeric antibodies as control.","FIG.","3 .","Kinetics of growth for murine B16 tumor in mice immunized with the immunotherapeutic combinations, specifically the idiotypic vaccine comprising the Ab2 MAb1E10 and of the GM3-VSSP ganglioside vaccine, as it is detailed in the Example 3.FIG.","4 : Evolution of skin metastases in melanoma patient.","The pictures were taken previous to the immunization with the vaccine NeuGcGM3/VSSP/ISA 51, 2 and 4 months after the treatment.","Uncolored halos around some lesions are observed, stabilized lesions, in one lesion a diminished size was observed and in one lesion increment was detected.","FIG.","5 : Evolution of lung metastases in a melanoma patient.","The lung right vertex observed by computer assisted axial tomography, the lesion size was 18×20 mm before the immunization with the vaccine NeuGcGM3/VSSP/ISA 51 and 4 months after the immunization stabilization of the lesion is observed.","FIG.","6 : Recognition of F(ab′)2 fragments from Ab2 MAb1E10 and from other MAbs with the same isotype by serum from patients immunized with the idiotypic vaccine containing Ab2 MAb1E10 and aluminum hydroxide gel as adjuvant.","FIG.","7 : Recognition of GM3(NeuGc) and GM3(NeuAc) gangliosides serum from patients immunized with the idiotypic vaccine containing Ab2 MAb1E10 and aluminum hydroxide gel as adjuvant.","detailed-description description=\"Detailed Description\" end=\"tail\"?"],["TECHNICAL FIELD This invention relates to human medicine, and especially to therapeutic vaccines that induce an immune response to tumors that over-express gangliosides.","PRIOR ART Gangliosides are components of the plasmatic membranes of the most of mammalian cells.","Even when these glycosphingolipids are expressed in normal tissues, they are very attractive targets for immunotherapy due to the expression patterns during the malignant transformation of the cells (Hakomori, S. Ann.","Rev.","Biochem.","50; 1981:733-764; Hakomori, Cancer Res.","45; 1985:2405-2414; Irie, R. F., et al.","In: Therapeutic monoclonal antibodies.","Borrebaeck, C. A. K., and Larrick, J. W.","(Eds.).","M Stockton Press, 1990, pp.","75-94).","The saccharide nature of the gangliosides, together to the fact that they are self-antigens makes them very low immunogenic molecules.","Some strategies have been used to increase the immunogenicity of these antigens.","They are based on presentation of gangliosides to the immune system in a different molecular environment.","One of these strategies has been the use of conjugated vaccines in which the saccharide antigen is covalently linked to a carrier protein where the carrier is very immunogenic for T cells.","It makes possible to obtain a strong and long lasting antibody immune response.","Using this procedure has been possible rise IgG antibodies against such gangliosides, although the titers after the re-immunizations were smaller in comparison with those obtained in a classic immune response against thymus-independent antigens (Helling, F et al.","Cancer Res.","54; 1994:197-203; Helling, F et al.","Cancer Res.","55; 1995:2783-2788; Livingston P O et al.","Cancer Immunol Immunother 45; 1997:10-19).","New vaccine compositions have been reported to induce an immune response against N-acetylated and N-glycolylated gangliosides.","These vaccines are based on hydrophobic conjugation between gangliosides and very small size proteoliposomes (VSSP) obtained from the association of the Outer Membrane Protein Complex (OMPC) from Neisseria meningitidis, Gram-negative bacteria, with gangliosides (gangliosides/VSSP) (Estévez F. Et al.","Vaccine 18; 1999:190-197; U.S. Pat.","No.","5,788,985 y U.S. Pat.","No.","6,149,921).","The vaccines composed by GM3 ganglioside/VSSP or the (NeuGcGM3) ganglioside/VSSP have raised high IgM and IgG antibodies titers specific against GM3 and NeuGcGM3.Otherwise, vaccination with GM3/VSSP increased survival of mice bearing melanoma B16, it also reduced tumor volumes and it increased rejection rate to the subcutaneous transplant of the tumor (Alonso et al.","Int.","J Oncology 15; 1999: 59-66; Car, A. et al.","Melanoma Research, in press).","The idiotype network theory proposed by Jerne 1974 (Jerne, N. K. Ann.","Immunol.","125C; 1974:373-389), pointed out the immune system as an antibody network with a complex interaction pattern between them and several natural antigens, that interactions happen through the variable regions or idiotype (Id) and by interfacing idiotype-antiidiotype, the immune system is regulated.","Jerne's theory supported new strategies for cancer active immunotherapy (AI) based on the use of anti-idiotype vaccines.","The vaccination is carried out with an antibody (Ab1) that recognizes the tumor-associated antigen, which induces anti-idiotype antibodies (Ab2).","These anti-idiotype antibodies mimic the nominal antigen and in turn they generate anti-anti-idiotype antibodies (Ab3) against the tumor-associated antigen (Schmolling J, et al.","Hybridoma 14; 1995:183-186).","On the other hand, the induction of immune response against tumor associated antigens has been reported after vaccination with anti-Id antibodies (Ab2) (Raychauhuri, S. et al.","J. Immunol.","137; 1986:1743-1749; Raychauhuri, S. et al.","J. Immunol.","139; 1987:3902-3910; Bhattacharya-Chatterjee, M. et al.","J. Immunol.","139; 1987:1354-1360; Bhattacharya-Chatterjee, M. et al.","J. Immunol.","141; 1988:1398-1403; Herlyn, D. et al.","Intern.","Rev.","Immunol.","4; 1989:347-357; Chen, Z-J.","et al.","Cell.","Imm.","Immunother.","Cancer; 1990:351-359; Herlyn, D. et al.","In Vivo 5; 1991:615-624; Furuya, A. et al.","Anticancer Res.","12; 1992:27-32; Mittelman, A. et al.","Proc.","Natl.","Acad.","Sci.","USA 89; 1992:466-470; Durrant, L G. et al.","Cancer Res.","54; 1994:4837-4840; Mittelman, A. et al.","Cancer Res.","54; 1994:415-421; Schmitt, H. et al.","Hybridoma 13; 1994:389-396; Chakrobarty, M. et al.","J. Immunother.","18; 1995:95-103; Chakrobarty, M. et al.","Cancer Res.","55; 1995:1525-1530; Foon, K A. et al.","Clin.","Cancer Res.","1; 1995:1285-1294; Herlyn, D. et al.","Hybridoma 14; 1995:159-166; Sclebusch, H. et al.","Hybridoma 14; 1995:167-174; Herlyn, D. et al.","Cancer Immunol.","Immunother.","43; 1996:65-76).","Another choice, concerning the use of Ab2 antibodies and in order to increase the immunogenicity of saccharide residues, involves the possibility that such saccharide epitopo can be represented by a protein epitopo on the antibody molecule.","In fact, many of these anti-Id antibodies have been obtained; they mimic gangliosides highly expressed in tumor cells such as GM3, GD3 and GD2 (Yamamoto, S et al.","J. Natl.","Cancer Inst.","82; 1990:1757-1760; Chapman, P. B. et al.","J. Clin.","Invest 88; 1991:186-192; Cheung N-K V, et al.","Int J Cancer 54; 1993:499-505; Saleh M N. Et al.","J Immunol 151; 1993:3390-3398; Sen G. Et al.","J Immunotherapy 21; 1998: 75-83).","Promissory results have been achieved from clinical trials in cancer patients by using Ab2 antibodies simultaneously with BCG or QS21 as adjuvant (McCaffery M. Et al.","Clin Cancer Res 2; 1996: 679-686.; Foon K A.","Et al.","Clin Cancer Res.","4; 1998:1117-1124).","The monoclonal antibody P3 (Deposit Number ECACC 94113026) is known from the prior art, which specifically recognizes the sialic acid in N-glycolyl-containing monosialo and disialogangliosides.","This Mab Ab1 recognizes antigens on human breast tumors and melanomas.","(U.S. Pat.","No.","5,788,985; Vázquez A M. Et al.","Hybridoma 14; 1995:551-556; Moreno E. Et al.","Glycobiology 8; 1998: 695-705; Marquina G. Et al.","Cancer Res 56; 1996:5165-5171; Carr, A.","Et al.","Hybridoma 19 (3); 2000:241-247).","The anti-idiotype antibody 1E10 (Ab2 MAb 1E10) (Deposit Number ECACC 97112901) obtained by immunization with MAb P3 is a gamma type antibody (not internal image).","Ab2 MAb 1E10 showed anti-tumor effect on growth tumors of breast and melanomas (U.S. Pat.","No.","6,063,379; Vázquez et al.","Hybridoma 14; 1995:183-186; Vazquez et al.","Oncology Reports 7; 2000:751-756).","Do not exist evidences in the prior art about the use of the combination between vaccines comprising gangliosides and pharmaceutical compositions comprising anti-gangliosides antibodies (Ab1) or anti-idiotype antibodies (Ab2), named idiotypic vaccines; neither the use of combinations of idiotypic vaccine Ab1 with idiotypic vaccine Ab2.The present invention refers the use of all the possible vaccine combinations described, in order to potentiate the effect that they produce each one of them separately.","DETAILED DESCRIPTION OF THE INVENTION The present invention refers to a pharmaceutical composition or the combination of pharmaceutical compositions, useful for cancer immunotherapy, specifically for tumors that over express gangliosides, and that it comprises at least two of the following compounds: (A) An idiotypic vaccine comprising an anti-ganglioside antibody (Ab1); (B) An idiotypic vaccine comprising an anti-idiotype antibody (Ab2) against an anti-ganglioside antibody; and (C) A vaccine comprising a ganglioside.","The pharmaceutical composition, or the combination of pharmaceutical compositions can have A plus B, or A plus C, or B plus C. Preferably that pharmaceutical composition, or combination of the pharmaceutical compositions, where A can be an idiotypic vaccine comprising the murine MAb P3 with Deposit Number ECACC 94113026; and where B can be an idiotypic vaccine comprising the murine anti-idiotype antibody 1E10 with Deposit Number ECACC 97112901 and where C can be a vaccine comprising N-glycolyl GM3 (NeuGcGM3) or N-acetyl GM3 (NeuAcGM3) gangliosides.","In the present invention A plus B, or A plus C, or B plus C can be administered in a simultaneous or alternating way.","The compositions of this invention may be useful in the treatment of cancer, particularly those which over express gangliosides as well as lung, breast, digestive system, urogenital system, melanomas, sarcomas and those derived from neuroectodermic tissue.","The present invention also described the scheme for the administration of the pharmaceutical compositions, or combination of pharmaceutical compositions to treat mammalians.","In the present invention a method is also described a method that comprises the administration to mammals of the pharmaceutical composition, or combination of pharmaceutical compositions described previously for the prevention or treatment of tumors of breast, lung, digestive system, urogenital system, melanomas, sarcomas and of neuroectodermic origin.","1.Preparation of Vaccine Comprising Gangliosides: The vaccines comprising gangliosides are obtained according to the specification of Estevez and col in Vaccine 18,1999: 190-197 and in U.S. Pat.","No.","5,788,985 y U.S. Pat.","No.","6,149,921).","Very Small Size Proteoliposomes (VSSP) obtained from the association of the Outer Membrane Protein Complex (OMPC) from the Gram-negative bacteria strain, Neisseda meningitidis, with synthetic or natural gangliosides incorporated therein (VSSP-G).","The gangliosides could be (NeuGcGM3), GD3 or (NeuAcGM3).","2.Preparation of Idiotypic Vaccines: The pharmaceutical compositions are obtained according to the specifications of U.S. Pat.","No.","5,817,513 and U.S. Pat.","No.","6,063,379.The vaccine comprises murine anti-ganglioside monoclonal antibodies such anti-gangliósidos such as MAb P3 or murine anti-idiotype monoclonal antibodies such as Mab 1E10 that recognize anti-ganglioside monoclonal antibodies.","3.Immunotherapeutic Combinations that Potentiate the Immune Response and the Antitumor Effect of Ganglioside Vaccines and Idiotypic Vaccines in Animal Models: The referred procedures can be used in mice or any other species of mammals.","The components of the combination are the ganglioside vaccine and the idiotypic vaccine; they can be combined in different ways.","In one combination, the animals can be immunized with 3 to 10 doses in a range of 25 μg to 1 mg of the murine anti-ganglioside monoclonal antibody; the interval between doses can be 7 and 14 days.","During this period the animals receive among 3 to 10 dose in a range among 60 to 1000 μg of the ganglioside vaccine, the interval between doses can be 7 to 14 days.","In another combination, the animals can be immunized with 3 to 10 dose in a range of 25 μg to 1 mg of the murine anti-idiotypic antibody specific against an anti-ganglioside antibody; the interval between doses can be 7 to 14 days.","During this period the animals receive among 3 to 10 dose in a range among 60 to 1000 μg of the ganglioside vaccine, the interval between doses can be 7 to 14 days.","The administration of both types of vaccines can be simultaneous or alternating.","The vaccines can be formulated as separate products or as a vaccine composition when they are administered in a simultaneous way.","When vaccines are administered in an alternating way, the intervals between each type of vaccine can be 3-7 days.","The vaccines are administered in an adjuvant that can be aluminum hydroxide (62.5 μg-2.5 mg), Montanide ISA 51 (0.1-1.2 ml/doses), or any other appropriate adjuvant.","The total volume for each dose can be 10 μl-2 ml.","Vaccines comprising gangliosides can be administered intradermic, subcutaneous, intramuscular, intraperitoneal, intramucosal or their combinations.","The same administration routes can be used for vaccines comprising antibodies.","The immunotherapeutic combinations increase the antibody immune response as well as the cellular immune response in treated animals.","The time of appearance of local tumors, the tumor volume and survival of treated subjects is compared with the same parameters for the control group, in order to evaluate the effectiveness of immunotherapeutic combinations.","When comparing these parameters among treated and control groups it can be observed a shorter time of appearance of local tumors, a decrease of the tumor volume and an increase in survival for the group treated with therapeutic combinations of the present invention.","The control groups are not only those animals treated with the adjuvant, but also those groups treated with just only one vaccine instead of with their combinations.","4.Immunotherapeutic Combinations that Potentiate the Immune Response and the Antitumor Effect of Ganglioside Vaccines and Idiotypic Vaccines in Human Patients: The procedure before referred can also be applied to cancer patients in different clinical stages.","Particularly those tumors that over express gangliosides as well as lung, breast, digestive system, urogenital system, melanomas, sarcomas and those derived from neuroectodermic tissue In one combination, the patients can be immunized with 3 to 10 dose in a range of 0.1 to 5 mg of the murine anti-ganglioside monoclonal antibody; the interval between doses can be 7 and 14 days.","During this period the patients receive among 3 to 10 dose in a range among 60 to 1000 μg of the ganglioside vaccine, the interval between doses can be 7 to 14 days.","In a second combination, the patients can be immunized with 3 to 10 dose in a range of 0.1 to 5 mg of the murine anti-idiotypic antibody specific against an anti-ganglioside antibody; the interval between doses can be 7 to 14 days.","During this period the patients receive among 4 to 6 dose in a range among 60 to 1000 μg of the ganglioside vaccine, the interval between doses can be 7 to 14 days.","In a third combination, the patients can be immunized with the murine anti-ganglioside monoclonal antibody; doses, frequency and intervals can be the described previously.","During this period the patients receive among 3 to 10 dose in a range among 0.1 to 2 mg of the murine anti-idiotypic antibody specific against an anti-ganglioside antibody.","The administration of both types of vaccines can be simultaneous or alternating.","The vaccines can be formulated as separate products or as a vaccine composition when they are administered in a simultaneously.","When vaccines are administered in an alternating way, the intervals between each type of vaccine can be 3-7 days.","The vaccines are administered in an adjuvant that can be aluminum hydroxide (1-5 mg/doses), Montanide ISA 51 (0.6-1.2 ml/doses), or any other appropriate adjuvant.","The total volume for each dose can be 10 μl-2 ml.","Vaccines comprising gangliosides can be administered intradermic, subcutaneous, intramuscular, intraperitoneal, intramucosal or their combinations.","The same route can be used for vaccines comprising antibodies.","During vaccination, some biochemical parameters and antibodies titers are monitored by measurements in blood.","The frequency can be 1 week to 3 months.","Cellular immunity is studied using lymphocytes from the patients.","The extractions are carried out with a frequency oscillating from one week to three months.","Finally, the patients are re-immunized with both vaccines at the concentrations before mentioned; the intervals between doses can be 1-6 months.","They can be administered simultaneously or not and during 1 to 2 years.","The present vaccination scheme induces in patients an antibody and cellular immune response increased, and therefore to reduce tumor burden.","EXAMPLES Example 1 Activation of Anti-Idiotype T2 and Anti- Anti-Idiotype T3 Cells in the Syngeneic Model, Induced by Immunization with Murine MAB P3 Female Balb/c and nude mice of the same genetic background 6-8 weeks old, were immunized subcutaneously with 100 μg of murine MAb P3 (anti-N-glycolylated ganglioside, Deposit Number ECACC 94113026; Vázquez et al., Hybridoma 14; 1995: 551-558; Moreno et al.","Glycobiolgy 8; 1998: 695-705) and complete Freund's adjuvant (CFA).","Seven days later mice were re-immunized with 50 μg of the antibody in incomplete Freund's adjuvant (IFA).","At day 10, draining lymph nodes were collected and the lymphocytes were obtained by pressing with a syringe.","The cellular suspension was used in experiments of cellular proliferation, which was measured by 3H-thymidine incorporation.","In vitro lymphocyte proliferation was measured by cultivating lymphocyte suspension in the presence of increased concentrations (25 to 150 μg/mL) of the murine MAb P3 and its Ab2 MAb 1E10 (Number of Deposit ECACC 97112901).","Isotype-matched murine MAbs were used as controls.","Stimulation indexes equal to or higher than 3 where considered positive.","Lymphocytes obtained after the immunization of Balb/c mice with P3 MAb specifically proliferated in the presence of this MAb and this proliferation was not observed when the cells were cultured in the presence of the murine control MAb A3 (IgM) (Alfonso M. and col.: Hybridoma 14; 1995: 209-216).","This proliferation was dependent on the presence of T cells; because it was not obtained with lymph node cells from P3 immunized athymic nu/nu mice.","Immunization with the murine MAb P3 not only induced the proliferation of T cells against the idiotype of this Ab1 MAb (T2), but also induced the proliferation of anti-anti-idiotype T cells (T3) specific for the murine Ab2 MAb 1E10 (IgG1), and not against Mab C5 of the same isotype (IgG1) (FIG.","1).","Example 2 Activation of Anti-Idiotype (T2) and Anti-Anti-Idiotype (T3) Induced by Immunization with the Chimeric Antibody P3 Chimeric antibody P3, whose amino acid sequences of its variable regions of the heavy (VH) and light (VL) chains are respectively shown in FIGS.","8 and 9, was used to immunize female Balb/c mice, 6-8 weeks old.","For the first subcutaneous dose, 100 μg of the antibody emulsified in ACF was used.","Seven days later animals were re-immunized with 50 μg of the antibody emulsified in AIF.","At day 10, draining lymph nodes (LN) were collected and the LN cell lymphoproliferative capacity was analyzed by incubating in the presence of the murine and chimeric variants of antibodies P3 and Ab2 1E10, whose sequences of the variable regions are shown in the FIGS.","13 (heavy chain) and 14 (light chain).","C5 murine MAb and its chimeric variant were used as isotype-matched controls (Patent Application WO 97/33916 A1).","Lymphocytes from mice immunized with the chimeric antibody P3 specifically proliferated when cultivated in the presence of this antibody or the chimeric 1E10 MAb, and also when cultivated in the presence of the murine variants of these antibodies.","Specificity of proliferation was demonstrated by using isotype-matched control MAbs.","Therefore, chimeric P3 MAb maintains the ability of the murine variant to induce specific proliferation of anti-idiotype (T2) and anti-anti-idiotype (T3) T cells (FIG.","2).","Example 3 Reinforcement of the Anti-Tumor Effect of the GM3-VSSP/Montanide ISA 51 Vaccine by its Combination with a Vaccine Composed by the Murine MAb 1E10 Adsorbed in Aluminum Hydroxide A group of 6-8 weeks old C57BL/6 female mice was immunized subcutaneously at days 0, 14, 28 and 42 with 50 μg of murine 1E10 MAb adsorbed in aluminum hydroxide.","At days 7, 21, 35 and 49, mice received intramuscular doses with 120 μg of GM3-VSSP/Montanide ISA 51.Another group of mice was immunized in a similar way, but beginning the immunizations with the vaccine preparation containing GM3 ganglioside.","Control groups separately immunized at the same time intervals and with the same number of doses of each vaccine or only with phosphate-buffered saline solution were used in these experiments.","At day 63, mice in all groups were subcutaneously inoculated with 10 000 cells of the murine tumor B16.Tumor growth was evaluated in all groups.","Mixed Lineal Pattern statistical data analysis was used for comparison of tumor growth between all groups and the group control that only received PBS.","Vaccination with 1E10 MAb alone didn't protect mice against a challenge with 10,000 melanoma B16 cells.","VSSP-GM3 vaccine caused a retard of the tumor growth (p<0.05).","The combinations of GM3-VSSP and 1E10 MAb vaccines led to an increase in the protective effect observed with GM3-VSSP vaccine alone (p<0.05) (FIG.","3).","Example 4 Immunization Schedule of Cancer Patients with NeuGc-GM3/VSSP Ganglioside Vaccine Using Montanide ISA 51 as Adjuvant Aiming to demonstrate the safety and immunogenicity of the immunization with the NeuGc-GM3/VSSP vaccine using Montanide ISA 51 as adjuvant (U.S. Pat.","No.","5,788,985 and U.S. Pat.","No.","6,149,921), a clinical trial was carried out in which 20 patients with advanced malignant melanoma, which were not eligible for any other onco-specific treatment, were immunized with the vaccine.","The patients received 9 doses of the vaccine preparation containing 200 μg of GM3 ganglioside.","Doses were administered at days 0, 14, 28, 42, 56, 84, 112, 140, and 168.According to physician's criteria, patients received additional doses every 28 days after the sixth month until they completed one year of treatment.","Blood samples were obtained for routine biochemistry determinations and for evaluating serum titers of anti-NeuGcGM3 ganglioside antibodies at days 0, 14, 28, 56, 112, 168, 224, 280, and 332.Antibodies titers were measured by ELISA.","Serum dilutions were considered positive when anti-ganglioside OD values were equal or higher than 0.1 (referred to anti-methanol OD).","The toxicity of the vaccine NeuGcGM3-VSSP/Montanide ISA 51 consisted on erythema, local pain, induration in the place of injection and fever, and this allowed classification of the toxicity as mild, grade I/II according to the OMS criteria.","Patients developed specific antibody titers against NeuGcGM3 (between 1:80 and 1:2560).","Detected antibody isotype included IgG and IgM (all patients) and IgG (75% of the patients; Table I).","In Patient 01, which entered into the trial with a diagnosis of advanced malignant melanoma (Evolutionary Metastatic Disease, EMD), regression and stabilization of some cutaneous lesions was observed after two months of treatment, with uncolored halos around these lesions in which the presence of inflammatory infiltrate and necrosis was demonstrated by anatomopathologic studies in biopsies (FIG 4.).","In patient 02, with diagnosis of advanced malignant melanoma, a stabilization of lung metastases lesion was observed in the right vertex 18-20 mm, during at least 4 months (FIG.","5).","TABLE I Antibody titers against NeuGcGM3 in patients with advanced malignant melanoma immunized with the NeuGcGM3/VSSP/Montanide ISA 51 vaccine.","Patients Day IgM IgG IgA 01 0 0 0 0 14 640 80 320 28 320 160 320 56 1280 160 320 02 0 160 160 160 14 160 160 320 28 160 160 320 56 1280 320 320 03 0 80 320 0 14 320 320 0 28 640 320 0 56 640 640 0 04 0 0 0 0 14 160 80 160 28 1280 80 160 56 2560 640 2560 Example 5 Immunization Schedule in Cancer Patients with a Idiotypic Vaccine Containing 1E10 Ab2 Mab Aluminum Hydroxide-Precipitated Aiming to demonstrate the safety and immunogenicity of an idiotype vaccine containing the murine anti-idiotype MAb 1E10 (U.S. Pat.","No.","5,817,513 and U.S. Pat.","No.","6,063,379) and aluminum hydroxide as adjuvant, a clinical trial was carried out with 20 advanced malignant melanoma patients, which were not eligible for any other onco-specific treatment.","Patients received 4 doses of the vaccine, consisting in 2 mg of the 1E10 MAb.","Blood samples were obtained before the treatment and 14 days after each immunization for routine biochemistry determinations and for evaluating serum titers of antibodies against 1E10 MAb idiotype and NeuGc-GM3 ganglioside.","Antibody titers were measured by an ELISA assay considering positive values those equal or higher than 0.15.Vaccine administration to the patients produced mild toxicity, classified as degree I and II, according to the OMS.","In 16 of the 17 valuable patients strong IgG Ab3 antibody responses were elicited, detectable after receiving 2 or 3 doses of the vaccine.","The analysis of the specificity of this Ab3, showed a preferential recognition by patient's sera of 1E10 MAb, when compared with other isotype-matched control MAbs, which suggested the induction of idiotype-specific component in the antibody response to 1E10 MAb.","This was corroborated by the strong sera reactivity against the F(ab′)2 fragments of this MAb, with a median titer of 1:15000 (titers from 1:10000 to more than 1:100000), with little or no recognition of the F(ab′)2 fragments of the control MAbs used as controls.(FIG.","6).","The antibodies generated against the NeuGcGM3 were, in most of the cases, both IgM and IgG, with titers 1:4000 and 1:800, respectively (FIG.","7).","A patient with diagnosis of malignant melanoma and liver metastases was included in the clinical trial; after the treatment it was shown stabilization of the disease for 8 months and the patient's survival was 15 months.","Example 6 Combined Immunization with Anti-Idiotypic Vaccine Comprising Ab2 MAb1E10 and NeuGcGM3-VSSP Ganglioside Vaccine A melanoma patient with metastases, which had been submitted to monthly surgery, after the diagnosis, received 6 doses of the idiotypic vaccine containing Ab2 MAb1E10 and aluminum hydroxide gel (2 mg of the MAb per dose), days 0,14,28,42,56.During this period, also it was administered to the patient the ganglioside vaccine containing 200 ug NeuGcGM3-VSSP ganglioside and Montanide ISA 51 as adjuvant, days 7, 21, 35, 49, 63.After the patient received this immunization scheme, he was re-immunized monthly with both vaccines simultaneously during two month.","For the period of vaccination new lesions didn't appear and the patient condition was good.","Example 7 Recognition of Tumor Tissues by the MAb 14F7 NeuGcGM3 ganglioside is recognized by MAb 14F7 (patent application WO 99/40119).","Recognition of tumor tissues by the antibody is shown.","Formalin fixed tissue was included in paraffin.","The histology was evaluated in hematoxillin-eosin colored sections.","These sections were immune staining with the complex biotin-estreptavidin-peroxidase (Hsu, S. M. y Raine, L. 1981,J.","Histochem Cytochem., 29:1349-1353).","Briefly, the paraffin was removed and wet sections were treated with H2O2 3% (in methanol solution) for 30 minutes, to reduce endogenous peroxidase activity.","After incubation with MAb 14F7 (pure) for 1 hour at room temperature, biotinylated anti-mouse immunoglobulins and streptavidin-peroxidase complex were added (Dakopatts) at room temperature; between incubation times, the sections were washed with a Tris-HCl buffer solution.","The reaction (POD) it was developed with 5 mL of Tris-HCl, 0.005 mL buffer solution containing H2O2 to 30% and of 3-3 diaminobencidine 3 mg. After washing with hematoxillin-contrasted water (Mayer), the sections were covered with balm and covered.","The enzyme reaction renders a brown color.","The fresh biopsies from pathological tissues were obtained one hour after surgery.","All the tissues were washed with saline solution, and frozen in liquid nitrogen immediately and they were conserved frozen at −80° C. The frozen fragments were cut in a cryostat Leica at −25° C. Serial sections with 5 um were obtained, air dried and used immediately or conserved at −20° C. enveloped in aluminum paper; afterward the sections were fixed with para-formaldehyde 4% for 20 minutes.","In Table II it is shown the immunostaining with MAb 14F7 in several human tumors.","A strong membrane and cytoplasm staining is appreciated in more than 50% of tumor cells.","The dye was very intense in colon carcinomas, uterus, ovary, sarcomas, lymph nodes and brain metastases of breast cancer, as well as melanoma metastases.","TABLE II Tumor Immunostaining with MAb 14F7.Immunostaining Tumor (Positives/Total) Colon Carcinomas 9/9* Uterus Carcinomas 2/2* Ovary Carcinomas 2/2* Sarcomas 2/2* Breast Tumors and lymph 6/6* node metastases Melanoma metastases 2/2* *More than 50% of cells showed membrane and cytoplasm intensely immunostained.","BRIEF DESCRIPTION OF THE FIGURES FIG.","1.Mice Balb/c euthymic and athymic mice were immunized subcutaneously with 100 μg of the murine MAb P3 in CFA followed by a re-immunization with 50 μg of the MAb emulsified in IFA.","After three days, lymph nodes cells were collected and it was carried out the proliferation assay with different concentrations of the MAbs P3, A3, 1E10 and C5.FIG.","2.Mice Balb/c were immunized subcutaneously with 100 pg of the chimeric antibody P3 in CFA followed by a re-immunization with 50 μg of the antibody emulsified in IFA.","After three days, lymph nodes cells were collected and it was carried out the proliferation assay with different concentrations using murine and chimeric antibodies as control.","FIG.","3.Kinetics of growth for murine B16 tumor in mice immunized with the immunotherapeutic combinations, specifically the idiotypic vaccine comprising the Ab2 MAb1E10 and of the GM3-VSSP ganglioside vaccine, as it is detailed in the Example 3.FIG.","4: Evolution of skin metastases in melanoma patient.","The pictures were taken previous to the immunization with the vaccine NeuGcGM3/VSSP/ISA 51, 2 and 4 months after the treatment.","Uncolored halos around some lesions are observed, stabilized lesions, in one lesion a diminished size was observed and in one lesion increment was detected.","FIG.","5: Evolution of lung metastases in a melanoma patient.","The lung right vertex observed by computer assisted axial tomography, the lesion size was 18×20 mm before the immunization with the vaccine NeuGcGM3/VSSP/ISA 51 and 4 months after the immunization stabilization of the lesion is observed.","FIG.","6: Recognition of F(ab′)2 fragments from Ab2 MAb1E10 and from other MAbs with the same isotype by serum from patients immunized with the idiotypic vaccine containing Ab2 MAb1E10 and aluminum hydroxide gel as adjuvant.","FIG.","7: Recognition of GM3(NeuGc) and GM3(NeuAc) gangliosides serum from patients immunized with the idiotypic vaccine containing Ab2 MAb1E10 and aluminum hydroxide gel as adjuvant."]],"string":"[\n [\n \"Immunotherapeutic combinations for the treatment of tumours that overexpress gangliosides\",\n \"The present invention is related to the field of immunology and more specifically with immunotherapeutic combinations used for controlling growth and/or cell proliferation of tumors.\",\n \"By means of the present invention a therapeutic effect against tumors is observed by the combination of idiotypic vaccines whose principle active is an anti-ganglioside antibody (Ab1), with idiotypic vaccines whose principle active is anti-idiotype antibody (Ab2) obtained against an anti-ganglioside antibody, or vaccines whose principle active is one or more gangliosides.\",\n \"Combinations of these vaccines are described that cause a synergic therapeutic effect against tumors.\",\n \"The referred combinations can be applied to patients in different clinical conditions of tumors that express gangliósidos.\"\n ],\n [\n \"1.An immunotherapeutic combination for the immunotherapy of tumors that over-express gangliosides, comprising at least two of the following components: (A) An idiotypic vaccine comprising a murine anti-ganglioside monoclonal antibody (Ab1); (B) An idiotypic vaccine comprising an specific murine anti-idiotype monoclonal antibody (Ab2), against an anti-ganglioside monoclonal antibody; (C) A ganglioside vaccine.\",\n \"2.An immunotherapeutic combination according to claim 1, wherein said combination comprises A plus B or A plus C or B plus C. 3.An immunotherapeutic combination according to claim 1, wherein said vaccine A comprises the murine anti-ganglioside MAb P3 (Deposit Number ECACC 94113026).\",\n \"4.An immunotherapeutic combination according to claim 1, wherein said vaccine B comprises the murine anti-idiotype antibody 1E10 (Deposit Number ECACC 97112901).\",\n \"5.An immunotherapeutic combination according to claim 1, wherein said vaccine C comprises the NeuGcGM3 ganglioside.\",\n \"6.Immunotherapeutic combination according to claim 1, wherein said vaccine C comprises the NeuAcGM3 ganglioside.\",\n \"7.Immunotherapeutic combination according to claims 1, wherein A and B or A and C or B and C are administered simultaneously or alternately.\",\n \"8.Use of any of the immunotherapeutic combinations of the claim 1 for the treatment of tumors that over-express gangliosides.\",\n \"9.Use according to claim 8, for treatment of tumor of breast, lung, digestive system, urogenital system, melanomas, sarcomas and those from neuroectodermic tissues.\",\n \"10.A method for controlling growth and/or cell proliferation of tumors that over-express gangliosides, comprising the administration to a mammal of an immunotherapeutic combination of claim 7.11.Method according to claim 10, wherein said mammals are human.\"\n ],\n [\n \"<SOH> TECHNICAL FIELD <EOH>This invention relates to human medicine, and especially to therapeutic vaccines that induce an immune response to tumors that over-express gangliosides.\"\n ],\n [\n \"<SOH> BRIEF DESCRIPTION OF THE FIGURES <EOH>FIG.\",\n \"1 .\",\n \"Mice Balb/c euthymic and athymic mice were immunized subcutaneously with 100 μg of the murine MAb P3 in CFA followed by a re-immunization with 50 μg of the MAb emulsified in IFA.\",\n \"After three days, lymph nodes cells were collected and it was carried out the proliferation assay with different concentrations of the MAbs P3, A3, 1E10 and C5.FIG.\",\n \"2 .\",\n \"Mice Balb/c were immunized subcutaneously with 100 pg of the chimeric antibody P3 in CFA followed by a re-immunization with 50 μg of the antibody emulsified in IFA.\",\n \"After three days, lymph nodes cells were collected and it was carried out the proliferation assay with different concentrations using murine and chimeric antibodies as control.\",\n \"FIG.\",\n \"3 .\",\n \"Kinetics of growth for murine B16 tumor in mice immunized with the immunotherapeutic combinations, specifically the idiotypic vaccine comprising the Ab2 MAb1E10 and of the GM3-VSSP ganglioside vaccine, as it is detailed in the Example 3.FIG.\",\n \"4 : Evolution of skin metastases in melanoma patient.\",\n \"The pictures were taken previous to the immunization with the vaccine NeuGcGM3/VSSP/ISA 51, 2 and 4 months after the treatment.\",\n \"Uncolored halos around some lesions are observed, stabilized lesions, in one lesion a diminished size was observed and in one lesion increment was detected.\",\n \"FIG.\",\n \"5 : Evolution of lung metastases in a melanoma patient.\",\n \"The lung right vertex observed by computer assisted axial tomography, the lesion size was 18×20 mm before the immunization with the vaccine NeuGcGM3/VSSP/ISA 51 and 4 months after the immunization stabilization of the lesion is observed.\",\n \"FIG.\",\n \"6 : Recognition of F(ab′)2 fragments from Ab2 MAb1E10 and from other MAbs with the same isotype by serum from patients immunized with the idiotypic vaccine containing Ab2 MAb1E10 and aluminum hydroxide gel as adjuvant.\",\n \"FIG.\",\n \"7 : Recognition of GM3(NeuGc) and GM3(NeuAc) gangliosides serum from patients immunized with the idiotypic vaccine containing Ab2 MAb1E10 and aluminum hydroxide gel as adjuvant.\",\n \"detailed-description description=\\\"Detailed Description\\\" end=\\\"tail\\\"?\"\n ],\n [\n \"TECHNICAL FIELD This invention relates to human medicine, and especially to therapeutic vaccines that induce an immune response to tumors that over-express gangliosides.\",\n \"PRIOR ART Gangliosides are components of the plasmatic membranes of the most of mammalian cells.\",\n \"Even when these glycosphingolipids are expressed in normal tissues, they are very attractive targets for immunotherapy due to the expression patterns during the malignant transformation of the cells (Hakomori, S. Ann.\",\n \"Rev.\",\n \"Biochem.\",\n \"50; 1981:733-764; Hakomori, Cancer Res.\",\n \"45; 1985:2405-2414; Irie, R. F., et al.\",\n \"In: Therapeutic monoclonal antibodies.\",\n \"Borrebaeck, C. A. K., and Larrick, J. W.\",\n \"(Eds.).\",\n \"M Stockton Press, 1990, pp.\",\n \"75-94).\",\n \"The saccharide nature of the gangliosides, together to the fact that they are self-antigens makes them very low immunogenic molecules.\",\n \"Some strategies have been used to increase the immunogenicity of these antigens.\",\n \"They are based on presentation of gangliosides to the immune system in a different molecular environment.\",\n \"One of these strategies has been the use of conjugated vaccines in which the saccharide antigen is covalently linked to a carrier protein where the carrier is very immunogenic for T cells.\",\n \"It makes possible to obtain a strong and long lasting antibody immune response.\",\n \"Using this procedure has been possible rise IgG antibodies against such gangliosides, although the titers after the re-immunizations were smaller in comparison with those obtained in a classic immune response against thymus-independent antigens (Helling, F et al.\",\n \"Cancer Res.\",\n \"54; 1994:197-203; Helling, F et al.\",\n \"Cancer Res.\",\n \"55; 1995:2783-2788; Livingston P O et al.\",\n \"Cancer Immunol Immunother 45; 1997:10-19).\",\n \"New vaccine compositions have been reported to induce an immune response against N-acetylated and N-glycolylated gangliosides.\",\n \"These vaccines are based on hydrophobic conjugation between gangliosides and very small size proteoliposomes (VSSP) obtained from the association of the Outer Membrane Protein Complex (OMPC) from Neisseria meningitidis, Gram-negative bacteria, with gangliosides (gangliosides/VSSP) (Estévez F. Et al.\",\n \"Vaccine 18; 1999:190-197; U.S. Pat.\",\n \"No.\",\n \"5,788,985 y U.S. Pat.\",\n \"No.\",\n \"6,149,921).\",\n \"The vaccines composed by GM3 ganglioside/VSSP or the (NeuGcGM3) ganglioside/VSSP have raised high IgM and IgG antibodies titers specific against GM3 and NeuGcGM3.Otherwise, vaccination with GM3/VSSP increased survival of mice bearing melanoma B16, it also reduced tumor volumes and it increased rejection rate to the subcutaneous transplant of the tumor (Alonso et al.\",\n \"Int.\",\n \"J Oncology 15; 1999: 59-66; Car, A. et al.\",\n \"Melanoma Research, in press).\",\n \"The idiotype network theory proposed by Jerne 1974 (Jerne, N. K. Ann.\",\n \"Immunol.\",\n \"125C; 1974:373-389), pointed out the immune system as an antibody network with a complex interaction pattern between them and several natural antigens, that interactions happen through the variable regions or idiotype (Id) and by interfacing idiotype-antiidiotype, the immune system is regulated.\",\n \"Jerne's theory supported new strategies for cancer active immunotherapy (AI) based on the use of anti-idiotype vaccines.\",\n \"The vaccination is carried out with an antibody (Ab1) that recognizes the tumor-associated antigen, which induces anti-idiotype antibodies (Ab2).\",\n \"These anti-idiotype antibodies mimic the nominal antigen and in turn they generate anti-anti-idiotype antibodies (Ab3) against the tumor-associated antigen (Schmolling J, et al.\",\n \"Hybridoma 14; 1995:183-186).\",\n \"On the other hand, the induction of immune response against tumor associated antigens has been reported after vaccination with anti-Id antibodies (Ab2) (Raychauhuri, S. et al.\",\n \"J. Immunol.\",\n \"137; 1986:1743-1749; Raychauhuri, S. et al.\",\n \"J. Immunol.\",\n \"139; 1987:3902-3910; Bhattacharya-Chatterjee, M. et al.\",\n \"J. Immunol.\",\n \"139; 1987:1354-1360; Bhattacharya-Chatterjee, M. et al.\",\n \"J. Immunol.\",\n \"141; 1988:1398-1403; Herlyn, D. et al.\",\n \"Intern.\",\n \"Rev.\",\n \"Immunol.\",\n \"4; 1989:347-357; Chen, Z-J.\",\n \"et al.\",\n \"Cell.\",\n \"Imm.\",\n \"Immunother.\",\n \"Cancer; 1990:351-359; Herlyn, D. et al.\",\n \"In Vivo 5; 1991:615-624; Furuya, A. et al.\",\n \"Anticancer Res.\",\n \"12; 1992:27-32; Mittelman, A. et al.\",\n \"Proc.\",\n \"Natl.\",\n \"Acad.\",\n \"Sci.\",\n \"USA 89; 1992:466-470; Durrant, L G. et al.\",\n \"Cancer Res.\",\n \"54; 1994:4837-4840; Mittelman, A. et al.\",\n \"Cancer Res.\",\n \"54; 1994:415-421; Schmitt, H. et al.\",\n \"Hybridoma 13; 1994:389-396; Chakrobarty, M. et al.\",\n \"J. Immunother.\",\n \"18; 1995:95-103; Chakrobarty, M. et al.\",\n \"Cancer Res.\",\n \"55; 1995:1525-1530; Foon, K A. et al.\",\n \"Clin.\",\n \"Cancer Res.\",\n \"1; 1995:1285-1294; Herlyn, D. et al.\",\n \"Hybridoma 14; 1995:159-166; Sclebusch, H. et al.\",\n \"Hybridoma 14; 1995:167-174; Herlyn, D. et al.\",\n \"Cancer Immunol.\",\n \"Immunother.\",\n \"43; 1996:65-76).\",\n \"Another choice, concerning the use of Ab2 antibodies and in order to increase the immunogenicity of saccharide residues, involves the possibility that such saccharide epitopo can be represented by a protein epitopo on the antibody molecule.\",\n \"In fact, many of these anti-Id antibodies have been obtained; they mimic gangliosides highly expressed in tumor cells such as GM3, GD3 and GD2 (Yamamoto, S et al.\",\n \"J. Natl.\",\n \"Cancer Inst.\",\n \"82; 1990:1757-1760; Chapman, P. B. et al.\",\n \"J. Clin.\",\n \"Invest 88; 1991:186-192; Cheung N-K V, et al.\",\n \"Int J Cancer 54; 1993:499-505; Saleh M N. Et al.\",\n \"J Immunol 151; 1993:3390-3398; Sen G. Et al.\",\n \"J Immunotherapy 21; 1998: 75-83).\",\n \"Promissory results have been achieved from clinical trials in cancer patients by using Ab2 antibodies simultaneously with BCG or QS21 as adjuvant (McCaffery M. Et al.\",\n \"Clin Cancer Res 2; 1996: 679-686.; Foon K A.\",\n \"Et al.\",\n \"Clin Cancer Res.\",\n \"4; 1998:1117-1124).\",\n \"The monoclonal antibody P3 (Deposit Number ECACC 94113026) is known from the prior art, which specifically recognizes the sialic acid in N-glycolyl-containing monosialo and disialogangliosides.\",\n \"This Mab Ab1 recognizes antigens on human breast tumors and melanomas.\",\n \"(U.S. Pat.\",\n \"No.\",\n \"5,788,985; Vázquez A M. Et al.\",\n \"Hybridoma 14; 1995:551-556; Moreno E. Et al.\",\n \"Glycobiology 8; 1998: 695-705; Marquina G. Et al.\",\n \"Cancer Res 56; 1996:5165-5171; Carr, A.\",\n \"Et al.\",\n \"Hybridoma 19 (3); 2000:241-247).\",\n \"The anti-idiotype antibody 1E10 (Ab2 MAb 1E10) (Deposit Number ECACC 97112901) obtained by immunization with MAb P3 is a gamma type antibody (not internal image).\",\n \"Ab2 MAb 1E10 showed anti-tumor effect on growth tumors of breast and melanomas (U.S. Pat.\",\n \"No.\",\n \"6,063,379; Vázquez et al.\",\n \"Hybridoma 14; 1995:183-186; Vazquez et al.\",\n \"Oncology Reports 7; 2000:751-756).\",\n \"Do not exist evidences in the prior art about the use of the combination between vaccines comprising gangliosides and pharmaceutical compositions comprising anti-gangliosides antibodies (Ab1) or anti-idiotype antibodies (Ab2), named idiotypic vaccines; neither the use of combinations of idiotypic vaccine Ab1 with idiotypic vaccine Ab2.The present invention refers the use of all the possible vaccine combinations described, in order to potentiate the effect that they produce each one of them separately.\",\n \"DETAILED DESCRIPTION OF THE INVENTION The present invention refers to a pharmaceutical composition or the combination of pharmaceutical compositions, useful for cancer immunotherapy, specifically for tumors that over express gangliosides, and that it comprises at least two of the following compounds: (A) An idiotypic vaccine comprising an anti-ganglioside antibody (Ab1); (B) An idiotypic vaccine comprising an anti-idiotype antibody (Ab2) against an anti-ganglioside antibody; and (C) A vaccine comprising a ganglioside.\",\n \"The pharmaceutical composition, or the combination of pharmaceutical compositions can have A plus B, or A plus C, or B plus C. Preferably that pharmaceutical composition, or combination of the pharmaceutical compositions, where A can be an idiotypic vaccine comprising the murine MAb P3 with Deposit Number ECACC 94113026; and where B can be an idiotypic vaccine comprising the murine anti-idiotype antibody 1E10 with Deposit Number ECACC 97112901 and where C can be a vaccine comprising N-glycolyl GM3 (NeuGcGM3) or N-acetyl GM3 (NeuAcGM3) gangliosides.\",\n \"In the present invention A plus B, or A plus C, or B plus C can be administered in a simultaneous or alternating way.\",\n \"The compositions of this invention may be useful in the treatment of cancer, particularly those which over express gangliosides as well as lung, breast, digestive system, urogenital system, melanomas, sarcomas and those derived from neuroectodermic tissue.\",\n \"The present invention also described the scheme for the administration of the pharmaceutical compositions, or combination of pharmaceutical compositions to treat mammalians.\",\n \"In the present invention a method is also described a method that comprises the administration to mammals of the pharmaceutical composition, or combination of pharmaceutical compositions described previously for the prevention or treatment of tumors of breast, lung, digestive system, urogenital system, melanomas, sarcomas and of neuroectodermic origin.\",\n \"1.Preparation of Vaccine Comprising Gangliosides: The vaccines comprising gangliosides are obtained according to the specification of Estevez and col in Vaccine 18,1999: 190-197 and in U.S. Pat.\",\n \"No.\",\n \"5,788,985 y U.S. Pat.\",\n \"No.\",\n \"6,149,921).\",\n \"Very Small Size Proteoliposomes (VSSP) obtained from the association of the Outer Membrane Protein Complex (OMPC) from the Gram-negative bacteria strain, Neisseda meningitidis, with synthetic or natural gangliosides incorporated therein (VSSP-G).\",\n \"The gangliosides could be (NeuGcGM3), GD3 or (NeuAcGM3).\",\n \"2.Preparation of Idiotypic Vaccines: The pharmaceutical compositions are obtained according to the specifications of U.S. Pat.\",\n \"No.\",\n \"5,817,513 and U.S. Pat.\",\n \"No.\",\n \"6,063,379.The vaccine comprises murine anti-ganglioside monoclonal antibodies such anti-gangliósidos such as MAb P3 or murine anti-idiotype monoclonal antibodies such as Mab 1E10 that recognize anti-ganglioside monoclonal antibodies.\",\n \"3.Immunotherapeutic Combinations that Potentiate the Immune Response and the Antitumor Effect of Ganglioside Vaccines and Idiotypic Vaccines in Animal Models: The referred procedures can be used in mice or any other species of mammals.\",\n \"The components of the combination are the ganglioside vaccine and the idiotypic vaccine; they can be combined in different ways.\",\n \"In one combination, the animals can be immunized with 3 to 10 doses in a range of 25 μg to 1 mg of the murine anti-ganglioside monoclonal antibody; the interval between doses can be 7 and 14 days.\",\n \"During this period the animals receive among 3 to 10 dose in a range among 60 to 1000 μg of the ganglioside vaccine, the interval between doses can be 7 to 14 days.\",\n \"In another combination, the animals can be immunized with 3 to 10 dose in a range of 25 μg to 1 mg of the murine anti-idiotypic antibody specific against an anti-ganglioside antibody; the interval between doses can be 7 to 14 days.\",\n \"During this period the animals receive among 3 to 10 dose in a range among 60 to 1000 μg of the ganglioside vaccine, the interval between doses can be 7 to 14 days.\",\n \"The administration of both types of vaccines can be simultaneous or alternating.\",\n \"The vaccines can be formulated as separate products or as a vaccine composition when they are administered in a simultaneous way.\",\n \"When vaccines are administered in an alternating way, the intervals between each type of vaccine can be 3-7 days.\",\n \"The vaccines are administered in an adjuvant that can be aluminum hydroxide (62.5 μg-2.5 mg), Montanide ISA 51 (0.1-1.2 ml/doses), or any other appropriate adjuvant.\",\n \"The total volume for each dose can be 10 μl-2 ml.\",\n \"Vaccines comprising gangliosides can be administered intradermic, subcutaneous, intramuscular, intraperitoneal, intramucosal or their combinations.\",\n \"The same administration routes can be used for vaccines comprising antibodies.\",\n \"The immunotherapeutic combinations increase the antibody immune response as well as the cellular immune response in treated animals.\",\n \"The time of appearance of local tumors, the tumor volume and survival of treated subjects is compared with the same parameters for the control group, in order to evaluate the effectiveness of immunotherapeutic combinations.\",\n \"When comparing these parameters among treated and control groups it can be observed a shorter time of appearance of local tumors, a decrease of the tumor volume and an increase in survival for the group treated with therapeutic combinations of the present invention.\",\n \"The control groups are not only those animals treated with the adjuvant, but also those groups treated with just only one vaccine instead of with their combinations.\",\n \"4.Immunotherapeutic Combinations that Potentiate the Immune Response and the Antitumor Effect of Ganglioside Vaccines and Idiotypic Vaccines in Human Patients: The procedure before referred can also be applied to cancer patients in different clinical stages.\",\n \"Particularly those tumors that over express gangliosides as well as lung, breast, digestive system, urogenital system, melanomas, sarcomas and those derived from neuroectodermic tissue In one combination, the patients can be immunized with 3 to 10 dose in a range of 0.1 to 5 mg of the murine anti-ganglioside monoclonal antibody; the interval between doses can be 7 and 14 days.\",\n \"During this period the patients receive among 3 to 10 dose in a range among 60 to 1000 μg of the ganglioside vaccine, the interval between doses can be 7 to 14 days.\",\n \"In a second combination, the patients can be immunized with 3 to 10 dose in a range of 0.1 to 5 mg of the murine anti-idiotypic antibody specific against an anti-ganglioside antibody; the interval between doses can be 7 to 14 days.\",\n \"During this period the patients receive among 4 to 6 dose in a range among 60 to 1000 μg of the ganglioside vaccine, the interval between doses can be 7 to 14 days.\",\n \"In a third combination, the patients can be immunized with the murine anti-ganglioside monoclonal antibody; doses, frequency and intervals can be the described previously.\",\n \"During this period the patients receive among 3 to 10 dose in a range among 0.1 to 2 mg of the murine anti-idiotypic antibody specific against an anti-ganglioside antibody.\",\n \"The administration of both types of vaccines can be simultaneous or alternating.\",\n \"The vaccines can be formulated as separate products or as a vaccine composition when they are administered in a simultaneously.\",\n \"When vaccines are administered in an alternating way, the intervals between each type of vaccine can be 3-7 days.\",\n \"The vaccines are administered in an adjuvant that can be aluminum hydroxide (1-5 mg/doses), Montanide ISA 51 (0.6-1.2 ml/doses), or any other appropriate adjuvant.\",\n \"The total volume for each dose can be 10 μl-2 ml.\",\n \"Vaccines comprising gangliosides can be administered intradermic, subcutaneous, intramuscular, intraperitoneal, intramucosal or their combinations.\",\n \"The same route can be used for vaccines comprising antibodies.\",\n \"During vaccination, some biochemical parameters and antibodies titers are monitored by measurements in blood.\",\n \"The frequency can be 1 week to 3 months.\",\n \"Cellular immunity is studied using lymphocytes from the patients.\",\n \"The extractions are carried out with a frequency oscillating from one week to three months.\",\n \"Finally, the patients are re-immunized with both vaccines at the concentrations before mentioned; the intervals between doses can be 1-6 months.\",\n \"They can be administered simultaneously or not and during 1 to 2 years.\",\n \"The present vaccination scheme induces in patients an antibody and cellular immune response increased, and therefore to reduce tumor burden.\",\n \"EXAMPLES Example 1 Activation of Anti-Idiotype T2 and Anti- Anti-Idiotype T3 Cells in the Syngeneic Model, Induced by Immunization with Murine MAB P3 Female Balb/c and nude mice of the same genetic background 6-8 weeks old, were immunized subcutaneously with 100 μg of murine MAb P3 (anti-N-glycolylated ganglioside, Deposit Number ECACC 94113026; Vázquez et al., Hybridoma 14; 1995: 551-558; Moreno et al.\",\n \"Glycobiolgy 8; 1998: 695-705) and complete Freund's adjuvant (CFA).\",\n \"Seven days later mice were re-immunized with 50 μg of the antibody in incomplete Freund's adjuvant (IFA).\",\n \"At day 10, draining lymph nodes were collected and the lymphocytes were obtained by pressing with a syringe.\",\n \"The cellular suspension was used in experiments of cellular proliferation, which was measured by 3H-thymidine incorporation.\",\n \"In vitro lymphocyte proliferation was measured by cultivating lymphocyte suspension in the presence of increased concentrations (25 to 150 μg/mL) of the murine MAb P3 and its Ab2 MAb 1E10 (Number of Deposit ECACC 97112901).\",\n \"Isotype-matched murine MAbs were used as controls.\",\n \"Stimulation indexes equal to or higher than 3 where considered positive.\",\n \"Lymphocytes obtained after the immunization of Balb/c mice with P3 MAb specifically proliferated in the presence of this MAb and this proliferation was not observed when the cells were cultured in the presence of the murine control MAb A3 (IgM) (Alfonso M. and col.: Hybridoma 14; 1995: 209-216).\",\n \"This proliferation was dependent on the presence of T cells; because it was not obtained with lymph node cells from P3 immunized athymic nu/nu mice.\",\n \"Immunization with the murine MAb P3 not only induced the proliferation of T cells against the idiotype of this Ab1 MAb (T2), but also induced the proliferation of anti-anti-idiotype T cells (T3) specific for the murine Ab2 MAb 1E10 (IgG1), and not against Mab C5 of the same isotype (IgG1) (FIG.\",\n \"1).\",\n \"Example 2 Activation of Anti-Idiotype (T2) and Anti-Anti-Idiotype (T3) Induced by Immunization with the Chimeric Antibody P3 Chimeric antibody P3, whose amino acid sequences of its variable regions of the heavy (VH) and light (VL) chains are respectively shown in FIGS.\",\n \"8 and 9, was used to immunize female Balb/c mice, 6-8 weeks old.\",\n \"For the first subcutaneous dose, 100 μg of the antibody emulsified in ACF was used.\",\n \"Seven days later animals were re-immunized with 50 μg of the antibody emulsified in AIF.\",\n \"At day 10, draining lymph nodes (LN) were collected and the LN cell lymphoproliferative capacity was analyzed by incubating in the presence of the murine and chimeric variants of antibodies P3 and Ab2 1E10, whose sequences of the variable regions are shown in the FIGS.\",\n \"13 (heavy chain) and 14 (light chain).\",\n \"C5 murine MAb and its chimeric variant were used as isotype-matched controls (Patent Application WO 97/33916 A1).\",\n \"Lymphocytes from mice immunized with the chimeric antibody P3 specifically proliferated when cultivated in the presence of this antibody or the chimeric 1E10 MAb, and also when cultivated in the presence of the murine variants of these antibodies.\",\n \"Specificity of proliferation was demonstrated by using isotype-matched control MAbs.\",\n \"Therefore, chimeric P3 MAb maintains the ability of the murine variant to induce specific proliferation of anti-idiotype (T2) and anti-anti-idiotype (T3) T cells (FIG.\",\n \"2).\",\n \"Example 3 Reinforcement of the Anti-Tumor Effect of the GM3-VSSP/Montanide ISA 51 Vaccine by its Combination with a Vaccine Composed by the Murine MAb 1E10 Adsorbed in Aluminum Hydroxide A group of 6-8 weeks old C57BL/6 female mice was immunized subcutaneously at days 0, 14, 28 and 42 with 50 μg of murine 1E10 MAb adsorbed in aluminum hydroxide.\",\n \"At days 7, 21, 35 and 49, mice received intramuscular doses with 120 μg of GM3-VSSP/Montanide ISA 51.Another group of mice was immunized in a similar way, but beginning the immunizations with the vaccine preparation containing GM3 ganglioside.\",\n \"Control groups separately immunized at the same time intervals and with the same number of doses of each vaccine or only with phosphate-buffered saline solution were used in these experiments.\",\n \"At day 63, mice in all groups were subcutaneously inoculated with 10 000 cells of the murine tumor B16.Tumor growth was evaluated in all groups.\",\n \"Mixed Lineal Pattern statistical data analysis was used for comparison of tumor growth between all groups and the group control that only received PBS.\",\n \"Vaccination with 1E10 MAb alone didn't protect mice against a challenge with 10,000 melanoma B16 cells.\",\n \"VSSP-GM3 vaccine caused a retard of the tumor growth (p<0.05).\",\n \"The combinations of GM3-VSSP and 1E10 MAb vaccines led to an increase in the protective effect observed with GM3-VSSP vaccine alone (p<0.05) (FIG.\",\n \"3).\",\n \"Example 4 Immunization Schedule of Cancer Patients with NeuGc-GM3/VSSP Ganglioside Vaccine Using Montanide ISA 51 as Adjuvant Aiming to demonstrate the safety and immunogenicity of the immunization with the NeuGc-GM3/VSSP vaccine using Montanide ISA 51 as adjuvant (U.S. Pat.\",\n \"No.\",\n \"5,788,985 and U.S. Pat.\",\n \"No.\",\n \"6,149,921), a clinical trial was carried out in which 20 patients with advanced malignant melanoma, which were not eligible for any other onco-specific treatment, were immunized with the vaccine.\",\n \"The patients received 9 doses of the vaccine preparation containing 200 μg of GM3 ganglioside.\",\n \"Doses were administered at days 0, 14, 28, 42, 56, 84, 112, 140, and 168.According to physician's criteria, patients received additional doses every 28 days after the sixth month until they completed one year of treatment.\",\n \"Blood samples were obtained for routine biochemistry determinations and for evaluating serum titers of anti-NeuGcGM3 ganglioside antibodies at days 0, 14, 28, 56, 112, 168, 224, 280, and 332.Antibodies titers were measured by ELISA.\",\n \"Serum dilutions were considered positive when anti-ganglioside OD values were equal or higher than 0.1 (referred to anti-methanol OD).\",\n \"The toxicity of the vaccine NeuGcGM3-VSSP/Montanide ISA 51 consisted on erythema, local pain, induration in the place of injection and fever, and this allowed classification of the toxicity as mild, grade I/II according to the OMS criteria.\",\n \"Patients developed specific antibody titers against NeuGcGM3 (between 1:80 and 1:2560).\",\n \"Detected antibody isotype included IgG and IgM (all patients) and IgG (75% of the patients; Table I).\",\n \"In Patient 01, which entered into the trial with a diagnosis of advanced malignant melanoma (Evolutionary Metastatic Disease, EMD), regression and stabilization of some cutaneous lesions was observed after two months of treatment, with uncolored halos around these lesions in which the presence of inflammatory infiltrate and necrosis was demonstrated by anatomopathologic studies in biopsies (FIG 4.).\",\n \"In patient 02, with diagnosis of advanced malignant melanoma, a stabilization of lung metastases lesion was observed in the right vertex 18-20 mm, during at least 4 months (FIG.\",\n \"5).\",\n \"TABLE I Antibody titers against NeuGcGM3 in patients with advanced malignant melanoma immunized with the NeuGcGM3/VSSP/Montanide ISA 51 vaccine.\",\n \"Patients Day IgM IgG IgA 01 0 0 0 0 14 640 80 320 28 320 160 320 56 1280 160 320 02 0 160 160 160 14 160 160 320 28 160 160 320 56 1280 320 320 03 0 80 320 0 14 320 320 0 28 640 320 0 56 640 640 0 04 0 0 0 0 14 160 80 160 28 1280 80 160 56 2560 640 2560 Example 5 Immunization Schedule in Cancer Patients with a Idiotypic Vaccine Containing 1E10 Ab2 Mab Aluminum Hydroxide-Precipitated Aiming to demonstrate the safety and immunogenicity of an idiotype vaccine containing the murine anti-idiotype MAb 1E10 (U.S. Pat.\",\n \"No.\",\n \"5,817,513 and U.S. Pat.\",\n \"No.\",\n \"6,063,379) and aluminum hydroxide as adjuvant, a clinical trial was carried out with 20 advanced malignant melanoma patients, which were not eligible for any other onco-specific treatment.\",\n \"Patients received 4 doses of the vaccine, consisting in 2 mg of the 1E10 MAb.\",\n \"Blood samples were obtained before the treatment and 14 days after each immunization for routine biochemistry determinations and for evaluating serum titers of antibodies against 1E10 MAb idiotype and NeuGc-GM3 ganglioside.\",\n \"Antibody titers were measured by an ELISA assay considering positive values those equal or higher than 0.15.Vaccine administration to the patients produced mild toxicity, classified as degree I and II, according to the OMS.\",\n \"In 16 of the 17 valuable patients strong IgG Ab3 antibody responses were elicited, detectable after receiving 2 or 3 doses of the vaccine.\",\n \"The analysis of the specificity of this Ab3, showed a preferential recognition by patient's sera of 1E10 MAb, when compared with other isotype-matched control MAbs, which suggested the induction of idiotype-specific component in the antibody response to 1E10 MAb.\",\n \"This was corroborated by the strong sera reactivity against the F(ab′)2 fragments of this MAb, with a median titer of 1:15000 (titers from 1:10000 to more than 1:100000), with little or no recognition of the F(ab′)2 fragments of the control MAbs used as controls.(FIG.\",\n \"6).\",\n \"The antibodies generated against the NeuGcGM3 were, in most of the cases, both IgM and IgG, with titers 1:4000 and 1:800, respectively (FIG.\",\n \"7).\",\n \"A patient with diagnosis of malignant melanoma and liver metastases was included in the clinical trial; after the treatment it was shown stabilization of the disease for 8 months and the patient's survival was 15 months.\",\n \"Example 6 Combined Immunization with Anti-Idiotypic Vaccine Comprising Ab2 MAb1E10 and NeuGcGM3-VSSP Ganglioside Vaccine A melanoma patient with metastases, which had been submitted to monthly surgery, after the diagnosis, received 6 doses of the idiotypic vaccine containing Ab2 MAb1E10 and aluminum hydroxide gel (2 mg of the MAb per dose), days 0,14,28,42,56.During this period, also it was administered to the patient the ganglioside vaccine containing 200 ug NeuGcGM3-VSSP ganglioside and Montanide ISA 51 as adjuvant, days 7, 21, 35, 49, 63.After the patient received this immunization scheme, he was re-immunized monthly with both vaccines simultaneously during two month.\",\n \"For the period of vaccination new lesions didn't appear and the patient condition was good.\",\n \"Example 7 Recognition of Tumor Tissues by the MAb 14F7 NeuGcGM3 ganglioside is recognized by MAb 14F7 (patent application WO 99/40119).\",\n \"Recognition of tumor tissues by the antibody is shown.\",\n \"Formalin fixed tissue was included in paraffin.\",\n \"The histology was evaluated in hematoxillin-eosin colored sections.\",\n \"These sections were immune staining with the complex biotin-estreptavidin-peroxidase (Hsu, S. M. y Raine, L. 1981,J.\",\n \"Histochem Cytochem., 29:1349-1353).\",\n \"Briefly, the paraffin was removed and wet sections were treated with H2O2 3% (in methanol solution) for 30 minutes, to reduce endogenous peroxidase activity.\",\n \"After incubation with MAb 14F7 (pure) for 1 hour at room temperature, biotinylated anti-mouse immunoglobulins and streptavidin-peroxidase complex were added (Dakopatts) at room temperature; between incubation times, the sections were washed with a Tris-HCl buffer solution.\",\n \"The reaction (POD) it was developed with 5 mL of Tris-HCl, 0.005 mL buffer solution containing H2O2 to 30% and of 3-3 diaminobencidine 3 mg. After washing with hematoxillin-contrasted water (Mayer), the sections were covered with balm and covered.\",\n \"The enzyme reaction renders a brown color.\",\n \"The fresh biopsies from pathological tissues were obtained one hour after surgery.\",\n \"All the tissues were washed with saline solution, and frozen in liquid nitrogen immediately and they were conserved frozen at −80° C. The frozen fragments were cut in a cryostat Leica at −25° C. Serial sections with 5 um were obtained, air dried and used immediately or conserved at −20° C. enveloped in aluminum paper; afterward the sections were fixed with para-formaldehyde 4% for 20 minutes.\",\n \"In Table II it is shown the immunostaining with MAb 14F7 in several human tumors.\",\n \"A strong membrane and cytoplasm staining is appreciated in more than 50% of tumor cells.\",\n \"The dye was very intense in colon carcinomas, uterus, ovary, sarcomas, lymph nodes and brain metastases of breast cancer, as well as melanoma metastases.\",\n \"TABLE II Tumor Immunostaining with MAb 14F7.Immunostaining Tumor (Positives/Total) Colon Carcinomas 9/9* Uterus Carcinomas 2/2* Ovary Carcinomas 2/2* Sarcomas 2/2* Breast Tumors and lymph 6/6* node metastases Melanoma metastases 2/2* *More than 50% of cells showed membrane and cytoplasm intensely immunostained.\",\n \"BRIEF DESCRIPTION OF THE FIGURES FIG.\",\n \"1.Mice Balb/c euthymic and athymic mice were immunized subcutaneously with 100 μg of the murine MAb P3 in CFA followed by a re-immunization with 50 μg of the MAb emulsified in IFA.\",\n \"After three days, lymph nodes cells were collected and it was carried out the proliferation assay with different concentrations of the MAbs P3, A3, 1E10 and C5.FIG.\",\n \"2.Mice Balb/c were immunized subcutaneously with 100 pg of the chimeric antibody P3 in CFA followed by a re-immunization with 50 μg of the antibody emulsified in IFA.\",\n \"After three days, lymph nodes cells were collected and it was carried out the proliferation assay with different concentrations using murine and chimeric antibodies as control.\",\n \"FIG.\",\n \"3.Kinetics of growth for murine B16 tumor in mice immunized with the immunotherapeutic combinations, specifically the idiotypic vaccine comprising the Ab2 MAb1E10 and of the GM3-VSSP ganglioside vaccine, as it is detailed in the Example 3.FIG.\",\n \"4: Evolution of skin metastases in melanoma patient.\",\n \"The pictures were taken previous to the immunization with the vaccine NeuGcGM3/VSSP/ISA 51, 2 and 4 months after the treatment.\",\n \"Uncolored halos around some lesions are observed, stabilized lesions, in one lesion a diminished size was observed and in one lesion increment was detected.\",\n \"FIG.\",\n \"5: Evolution of lung metastases in a melanoma patient.\",\n \"The lung right vertex observed by computer assisted axial tomography, the lesion size was 18×20 mm before the immunization with the vaccine NeuGcGM3/VSSP/ISA 51 and 4 months after the immunization stabilization of the lesion is observed.\",\n \"FIG.\",\n \"6: Recognition of F(ab′)2 fragments from Ab2 MAb1E10 and from other MAbs with the same isotype by serum from patients immunized with the idiotypic vaccine containing Ab2 MAb1E10 and aluminum hydroxide gel as adjuvant.\",\n \"FIG.\",\n \"7: Recognition of GM3(NeuGc) and GM3(NeuAc) gangliosides serum from patients immunized with the idiotypic vaccine containing Ab2 MAb1E10 and aluminum hydroxide gel as adjuvant.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473978"}}},{"rowIdx":776,"cells":{"text":{"kind":"list like","value":[["Device and method for data flow exchange between a client device and a server","The invention concerns a device for data flow exchange between a client device (3) provided with an Internet file and a server (4) adapted to provide on request from the client device (3), at least a service for transforming said file, the communication between the client (3) and the server (4) being based on a protocol of adaptation of Internet file content of the Internet Content Adaptation Protocol (iCAP) type.","The device comprises a adaptor (5) of iCAP type data flows into files adapted to be used directly by said server (4) or by said client device (3); the adaptor (5) comprises means for communicating with the client device (3) and the server (4) and configuration file wherein is/are stated the transformation service(s)."],["1.Device for exchanging flows between a client device (3) that has an Internet file and a server (4) able to provide, at the request of the client device (3), at least one service for converting said file, with communication between the client device (3) and the server (4) being based on an iCAP Internet file content adaptation protocol, characterised in that it comprises an adapter (5) of iCAP flows into files able to be used directly by said server (4) or said client device (3); and in that the adapter (5) includes means for communicating with the client device (3) and the server (4) and a configuration file in which the conversion service (services) is (are) declared.","2.Device according to claim 1, characterised in that the adapter (5) comprises a communication module (50) able to manage communication with the client device (3).","3.Device according to claim 1, characterised in that the adapter (5) comprises an Internet file header and content extraction module (51).","4.Device according to claim 3, characterised in that the adapter (5) comprises a header pre-analysis module (2).","5.Device according to claim 1, characterised in that the adapter (5) comprises at least one module (53a or 53b or 53c or 53d) for creating interface files between the adapter (5) and the server (4).","6.Device according to claim 1, characterised in that the adapter (5) comprises a module (54) is for calling up the adaptation service of the server (4).","7.Device according to claim 1, characterised in that the adapter (5) comprises a response module (55).","8.Device according to claim 1, characterised in that the adapter (5) comprises a module (56) for checking the integrity of the converted file intended for the client device (3).","9.Device according to claim 1, characterised in that the adapter (5) comprises a module (57) for restoring the converted file intended for the client device (3).","10.Device according to claim 1, characterised in that the adapter (5) is integrated with the server (4).","11.Method for exchanging flows between a client device that has an Internet file, and a server able to provide at least one file conversion service, with communication between the client and the server being based on an iCAP Internet file content adaptation protocol, characterised in that it consists in placing an adapter of iCAP flows into files able to be used between the client and the server and in that it comprises the following steps: a—the sending of an Internet file by said client device to the adapter, b—the adaptation of the Internet file by the adapter and the sending of the adapted file to said server, c—the execution of the conversion of the adapted file by said server and the sending of the converted file to said adapter, d—the adaptation of the converted file by the adapter e—the sending of the converted and adapted file to said client device.","12.Method according to claim 11, characterised in that step b—consists for the adapter in extracting the headers and possibly the content of the Internet file provided by the client device, pre-analysing the headers, generating interface files, calling up the server's conversion service, awaiting a response from the server.","13.Method according to claim 11, characterised in that step d—consists for the adapter in verifying the call return parameter and checking the integrity of the converted file, when the integrity check requests it, restoring the converted file.","14.Method according to claim 11, characterised in that it comprises a previous adapter initialisation step consisting in declaring the conversion service or services in an adapter configuration file."],["The present invention relates to a method for exchanging flows between a client device that has an Internet file and a server able to provide at the request of the client device a service for converting said file, communication between the client device and the server being based on an iCAP Internet file content adaptation protocol.","The field of the invention is that of communication between equipment in an Internet type telecommunication network and servers housing added value services, in the event content conversion or adaptation services.","In practice, content is presented in file form.","Communication is based on an Internet Content Adaptation Protocol defined by a group of companies in the context of the iCAP forum.","As examples of content conversion services may be cited file conversion services such as the conversion of Word documents into RTF documents, the search for viruses in a file, automatic file translation, the categorisation of content by semantic analysis returning a file corresponding to the main themes in the document, for the purpose for example of exercising parental control when licentious topics have been detected, etc.","An example of content conversion in accordance with the iCAP Protocol is shown in FIG.","1: a) an HTTP request is sent from the computer 1 of a user to a proxy cache 3 configured in the computer's browser; the computer 1 may be replaced by any other equipment able to send an HTTP request, likewise the cache 3 may be replaced by a router or by any other Internet network equipment, b) the cache 3 sends an HTTP request to the Web server 2 containing this page, c) the Web server returns the requested page to the cache 3, d) the cache 3 acting as the iCAP client device, known more straightforwardly as the iCAP client, requests a content conversion service from the iCAP Server 4, by transmitting the web page to it; the content conversion requested could also relate to the HTTP request itself and be requested directly, at the end of step a) or at the end of step c), in other words after the HTTP response has been sent; in the remainder of the document the term Internet file will be used to denote an HTTP response comprising one or more headers and a content, or an HTTP request not generally comprising content but comprising at least one header; e) the iCAP Server 4 processes this request by implementing the requested file conversion service then responds to the cache 3 by returning the converted file, for example the processed page, f) the cache 3 returns the response of the iCAP Server 4 to the user's computer 1 and stores this response.","As shown in this example, the role of the iCAP Protocol consists in connecting an iCAP Client 3 to the iCAP Server 4.At present in order to be able to implement this iCAP Protocol between a client 3 and a server 4 corresponding to the previous steps d) and e), it is |1 indispensable, as shown in FIG.","2, to develop a specific layer 41 within the file conversion service 40 implemented by the iCAP Server 4.This necessitates in particular a specific development for each file conversion service.","Moreover these developments are cumbersome and expensive.","The purpose of the present invention is therefore to reduce or even to eliminate the specific development that is required for each service to be compatible with the iCAP protocol.","The subject of the invention is a device for exchanging flows between a client who has an Internet file and a server able to provide, at the request of the client, at least one service for converting said file, with communication between the client and the server being based on an iCAP Internet file content adaptation protocol, mainly characterised in that it comprises an adapter of iCAP flows into files able to be used directly by said server or said client; and in that the adapter includes means: for communicating with the client and the server and a configuration file in which the conversion service (or services) is (or are) declared.","The adapter comprises to advantage a communication module able to manage communication with the client and/or a module for extracting headers and content from the Internet file and/or a module for pre-analysing headers and/or a module for creating interface files between the adapter and the server and/or a module for calling up the server's adaptation service and/or a response module and/or a module for checking the integrity of the converted file intended to the client and/or a module for restoring the converted file intended for the client.","The adapter may be integrated with the server.","The invention also relates to a method for exchanging flows between a client who has an Internet file, and a server able to provide at least one file conversion service, with communication between the client and the server being based on an iCAP Internet file content adaptation protocol, characterised in that it consists in placing an adapter of iCAP flows into files able to be used between the client and the server and in that it comprises the following steps: a—the sending of an Internet file, by said client to the adapter, b—the adaptation of the Internet file by the adapter and the sending of the adapted file to said server, c—the execution of the conversion of the adapted file, by said server and the sending of the converted file to said adapter, d—the adaptation of the converted file by the adapter e—the sending of the converted and adapted file to said client.","According to one characteristic of the invention, step b—consists for the adapter in extracting the headers and possibly the content of the Internet file provided by the client, pre-analysing the headers, generating interface files, calling up the server's conversion service, awaiting a response from the server.","Step d—consists to advantage for the adapter in verifying the call return parameter and checking the integrity of the converted file, when the integrity check requests it, restoring the converted file.","According to an additional characteristic, the method according to the invention comprises a previous step of initialising the adapter consisting in declaring the conversion service or services in an adapter configuration file.","Other particularities and advantages of the invention will emerge clearly from reading the description given by way of a non-restrictive example and in relation to the appended drawings in which: FIG.","1 already described shows diagrammatically the steps of calling upon a file conversion system, FIG.","2 already described shows diagrammatically the operation of a current file conversion system, FIG.","3 shows diagrammatically a file conversion system according to the invention, FIG.","4 shows diagrammatically the main operational elements of an iCAP flow exchange device according to the invention, FIG.","5 shows diagrammatically an example of an Internet file, FIGS.","6a), 6b), and 6c) show diagrammatically operational elements of a device according to the invention during conversion service call up, the execution of this service and the response respectively, FIG.","7 is a diagram of relationships between classes.","As shown in FIG.","3, the invention consists in providing an adapter 5 preferably in the server 4, so as, Ion the one hand in the client-server direction, to adapt files so that they may be directly used by the file conversion services 40 and, on the other hand in the server-client direction, to provide a check on the integrity of the converted file and possibly its restoration.","The adapter 5 makes it possible to formalise the exchanges between the iCAP client 3 and the file conversion services-by means of an adaptation of the iCAP flows in steps d) and e) in order to make them compatible with file conversion services 40 without it being necessary to develop an iCAP layer at file conversion service level.","This then makes it possible to define file conversion services that are modular, open-ended and independent of the iCAP layers.","The operation of the adapter according to the invention will now be described in relation to FIG.","4: the adapter 5 is firstly initialised in order particularly to know the conversion service or services that may be called upon: in a configuration file are 1entered moreover operational parameters (communication port, size of the conversion service request waiting queue, directory where interface files with the server are generated, etc), the names of the conversion services and the specific parameters associated with the conversion services and the way to invoke them (access path, service compilation model (using static or dynamic libraries or executables), etc.","); in one embodiment example, the operational parameters were initialised by the following fields and values: the communication port, server.default_port=8080 the size of the waiting queue, server.default_backlog=20 the directory, server.temp directory=/home/cbe/files the access path to specific files of a filter service, SVC.def_directory=/home/filtrage/SVC/def since it is initialised, the adapter 5 waits for the iCAP client 3 to connect via a communication module 50 thus allowing it to converse with the iCAP client 3 and to store temporarily all data exchanged during the communication phases.","When file conversion involves a response from the Web server 2, this data shown in FIG.","5 comprises, in addition to a content, the HTTP headers sent from the Web browser of the computer 1, the HTTP headers returned by the destination Web server, as well as the iCAP headers relating to the client 3.When file conversion involves a request, this data comprises the HTTP headers sent from the Web browser of the computer 1, as well as the iCAP headers relating to the iCAP client 3; the other data exists possibly in the event of the conversion being requested after step c) as shown in FIG.","1, but is not involved since it is a request conversion.","Reference is now made to FIG.","4.The module 50 manages the TCP/IP connection and the iCAP protocol in a conventional way by opening particularly a TCP communication port; this module 50 is in wait mode so long as no iCAP request reaches it.","When an iCAP request is received, the module sets up a new communication socket responsible for processing the conversion service requested by the client 3; the communication port so released returns to wait mode for another request.","The following written instruction in C++ language in the LINUX operating system may for example be used: The iCAPAdapter>handleNewClients( ); The data sent by the client 3 is transmitted to a module 500 via the communication socket that has been created.","The communication module 50 additionally comprises temporary storage means of all the data exchanged during the communication phases, by means of the module 500, the adapter 5 extracts the headers and the content from the data received from the module 50, analyses the headers, creates four files isolating the different headers from the content, using for example the instruction int iCr+CreateFiles(new_fd,cCanal,Exceed); after exact shaping the adapter 5 then redirects the data flow to the requested conversion service 40 using for example the instruction if(_ServiceIcap)returnValue-_ServiceIcap →CallService(new.fd,cCanal,_icapRootDirectory,Exceed); the service requested 40 may be localised on the server 4 or be remote and invoked through the Internet network for example.","The module 500 then puts itself into wait mode for a response from the conversion service; the converted file returned by the service 40 is checked by the module 500 which verifies its integrity; the converted file will possibly be subject to shaping, or even to a set of additional actions to make it comply with the iCAP protocol; the checked file is delivered to the iCAP client 3 via the communication module 50 and the communication socket opened to handle this conversion service is closed, using for example the instruction CiCapGlobals::ManageResponse(Filtrage,Long_file,root Directory,socket_fd);.","The diagram shown in FIG.","7 illustrates the relationships existing between the classes used in the previous instructions.","We will now describe in more detail the functionalities of the module 500 in relation to FIGS.","6a), 6b), and 6c) corresponding to a conversion service call, conversion execution and response respectively.","When a conversion service is called upon, the data is transmitted via the communication module 50 to a header and content extraction module 51.This module 51 separates the different fields specific to each of the headers and invokes the header pre-analysis module 52 for each header.","The header pre-analysis module 52 verifies initially that the conversion service invoked is available on the iCAP server; in the event, it verifies that the conversion service invoked is described in the adapter's configuration file.","If this is not the case, it returns an error to the iCAP client 3, via the communication module 50, without passing through the iCAP Server.","The module 52 also analyses the iCAP request type: Web response modification or Web request modification.","In request modification mode, it is not generally necessary to extract the content or even the HTTP header from the Web server.","The module 52 identifies any parameters associated with the execution of the service.","The module 52 returns the results to the extraction module 51 which calls on modules 53a, 53b, 53c, 53d to create output interface files.","IThe modules 53a, 53b, 53c, 53d create the four files offering the file conversion services 40 a common interface, which facilitates the conversions to be carried out on the files: the header of the iCAP server 4 is entered into an iCAP-Out file which then contains the information relating to the iCAP client 3 as well as a number of HTTP request or response fields depending on whether it is request modification mode or response modification mode, the HTTP header sent by the computer 1 which contains information relating to the client browser as well as to the Web server invoked is entered into an HTTPClient-out file, the HTTP header returned by the Web server 2 which contains information relating to the content of the response, its URL address, its type, its length is entered into an HTTPServ-out file; this file may be empty in the case of request modification node; the content of the Web server response, expurgated of the headers (html file, gif, jpeg, etc), is entered into an F-Content-Out file; this file may be empty in the case of request modification mode.","Each file name is completed by an index number incremented by one unit at each new iCAP request.","Each file creation module 53a, 53b, 53c, 53d sends a return code to the extraction module 51 to indicate the result of the operation, as well as the names of the files.","Once the headers are analysed and the files created, the extraction module 51 invokes the module 54 for calling up the conversion services 40.This module 54 calls up the relevant conversion service by providing a set of parameters comprising particularly the name of the service call function, the address and the iCAP server port where the service is located and the names of the previously defined interface files.","As shown in FIG.","6b), the conversion service 40 then executes the conversion in relation to modules 53a, 53b, 53c, 53d allowing the interface files to be accessed, and returns the converted data to a response module 55.The response module 55 shown in FIG.","6c), recovers the data from the conversion service 40.Among this data there features a service call function return code indicating that the conversion has been correctly carried out; when this is not the case and a problem has been detected, the return code, which is then an extended error code, allows the response module 55 to make a fast diagnosis of the problem detected.","Among this data there also features additional information in the form of four input interface files corresponding to the output interface files.","Each of these files reflects the file conversion that has occurred; the result is: an additional modification of the iCAP header, possible modifications of the client HTTP header, possible modifications of the server HTTP header, the data resulting from the converted file.","The response module 55 may use these files to call up a new iCAP service and thus link the conversion services together in so far as the data and parameters are available and already listed in the directory.","This functionality may be activated when the services to be linked together have the same generic name.","A virus checking service may for example follow a “parental” control service preventing access to Web pages for which licentious topics have been detected, these two services being recognised under the generic name “control”.","After receiving the converted data, the response module 55 invokes the integrity check module 56.The module 56 verifies the integrity of the data by carrying out particularly coherence tests between for example the length of the converted file and the header information relating to this length.","The checked data is then sent to the file restoration module 57.The module 57 creates or completes the iCAP header with the information on the nature of the iCAP server 4.It modifies the HTTP headers of the computer 1 and/or of the Web server 2 as a function of the data in the integrity check module 56; in the event of the absence of one or more input interface files, the module 57 may recreate them from the output interface files.","Once restored, the file is transmitted to the communication module 50, which returns it to the iCAP client 3 who has initialised the iCAP request."]],"string":"[\n [\n \"Device and method for data flow exchange between a client device and a server\",\n \"The invention concerns a device for data flow exchange between a client device (3) provided with an Internet file and a server (4) adapted to provide on request from the client device (3), at least a service for transforming said file, the communication between the client (3) and the server (4) being based on a protocol of adaptation of Internet file content of the Internet Content Adaptation Protocol (iCAP) type.\",\n \"The device comprises a adaptor (5) of iCAP type data flows into files adapted to be used directly by said server (4) or by said client device (3); the adaptor (5) comprises means for communicating with the client device (3) and the server (4) and configuration file wherein is/are stated the transformation service(s).\"\n ],\n [\n \"1.Device for exchanging flows between a client device (3) that has an Internet file and a server (4) able to provide, at the request of the client device (3), at least one service for converting said file, with communication between the client device (3) and the server (4) being based on an iCAP Internet file content adaptation protocol, characterised in that it comprises an adapter (5) of iCAP flows into files able to be used directly by said server (4) or said client device (3); and in that the adapter (5) includes means for communicating with the client device (3) and the server (4) and a configuration file in which the conversion service (services) is (are) declared.\",\n \"2.Device according to claim 1, characterised in that the adapter (5) comprises a communication module (50) able to manage communication with the client device (3).\",\n \"3.Device according to claim 1, characterised in that the adapter (5) comprises an Internet file header and content extraction module (51).\",\n \"4.Device according to claim 3, characterised in that the adapter (5) comprises a header pre-analysis module (2).\",\n \"5.Device according to claim 1, characterised in that the adapter (5) comprises at least one module (53a or 53b or 53c or 53d) for creating interface files between the adapter (5) and the server (4).\",\n \"6.Device according to claim 1, characterised in that the adapter (5) comprises a module (54) is for calling up the adaptation service of the server (4).\",\n \"7.Device according to claim 1, characterised in that the adapter (5) comprises a response module (55).\",\n \"8.Device according to claim 1, characterised in that the adapter (5) comprises a module (56) for checking the integrity of the converted file intended for the client device (3).\",\n \"9.Device according to claim 1, characterised in that the adapter (5) comprises a module (57) for restoring the converted file intended for the client device (3).\",\n \"10.Device according to claim 1, characterised in that the adapter (5) is integrated with the server (4).\",\n \"11.Method for exchanging flows between a client device that has an Internet file, and a server able to provide at least one file conversion service, with communication between the client and the server being based on an iCAP Internet file content adaptation protocol, characterised in that it consists in placing an adapter of iCAP flows into files able to be used between the client and the server and in that it comprises the following steps: a—the sending of an Internet file by said client device to the adapter, b—the adaptation of the Internet file by the adapter and the sending of the adapted file to said server, c—the execution of the conversion of the adapted file by said server and the sending of the converted file to said adapter, d—the adaptation of the converted file by the adapter e—the sending of the converted and adapted file to said client device.\",\n \"12.Method according to claim 11, characterised in that step b—consists for the adapter in extracting the headers and possibly the content of the Internet file provided by the client device, pre-analysing the headers, generating interface files, calling up the server's conversion service, awaiting a response from the server.\",\n \"13.Method according to claim 11, characterised in that step d—consists for the adapter in verifying the call return parameter and checking the integrity of the converted file, when the integrity check requests it, restoring the converted file.\",\n \"14.Method according to claim 11, characterised in that it comprises a previous adapter initialisation step consisting in declaring the conversion service or services in an adapter configuration file.\"\n ],\n [\n \"The present invention relates to a method for exchanging flows between a client device that has an Internet file and a server able to provide at the request of the client device a service for converting said file, communication between the client device and the server being based on an iCAP Internet file content adaptation protocol.\",\n \"The field of the invention is that of communication between equipment in an Internet type telecommunication network and servers housing added value services, in the event content conversion or adaptation services.\",\n \"In practice, content is presented in file form.\",\n \"Communication is based on an Internet Content Adaptation Protocol defined by a group of companies in the context of the iCAP forum.\",\n \"As examples of content conversion services may be cited file conversion services such as the conversion of Word documents into RTF documents, the search for viruses in a file, automatic file translation, the categorisation of content by semantic analysis returning a file corresponding to the main themes in the document, for the purpose for example of exercising parental control when licentious topics have been detected, etc.\",\n \"An example of content conversion in accordance with the iCAP Protocol is shown in FIG.\",\n \"1: a) an HTTP request is sent from the computer 1 of a user to a proxy cache 3 configured in the computer's browser; the computer 1 may be replaced by any other equipment able to send an HTTP request, likewise the cache 3 may be replaced by a router or by any other Internet network equipment, b) the cache 3 sends an HTTP request to the Web server 2 containing this page, c) the Web server returns the requested page to the cache 3, d) the cache 3 acting as the iCAP client device, known more straightforwardly as the iCAP client, requests a content conversion service from the iCAP Server 4, by transmitting the web page to it; the content conversion requested could also relate to the HTTP request itself and be requested directly, at the end of step a) or at the end of step c), in other words after the HTTP response has been sent; in the remainder of the document the term Internet file will be used to denote an HTTP response comprising one or more headers and a content, or an HTTP request not generally comprising content but comprising at least one header; e) the iCAP Server 4 processes this request by implementing the requested file conversion service then responds to the cache 3 by returning the converted file, for example the processed page, f) the cache 3 returns the response of the iCAP Server 4 to the user's computer 1 and stores this response.\",\n \"As shown in this example, the role of the iCAP Protocol consists in connecting an iCAP Client 3 to the iCAP Server 4.At present in order to be able to implement this iCAP Protocol between a client 3 and a server 4 corresponding to the previous steps d) and e), it is |1 indispensable, as shown in FIG.\",\n \"2, to develop a specific layer 41 within the file conversion service 40 implemented by the iCAP Server 4.This necessitates in particular a specific development for each file conversion service.\",\n \"Moreover these developments are cumbersome and expensive.\",\n \"The purpose of the present invention is therefore to reduce or even to eliminate the specific development that is required for each service to be compatible with the iCAP protocol.\",\n \"The subject of the invention is a device for exchanging flows between a client who has an Internet file and a server able to provide, at the request of the client, at least one service for converting said file, with communication between the client and the server being based on an iCAP Internet file content adaptation protocol, mainly characterised in that it comprises an adapter of iCAP flows into files able to be used directly by said server or said client; and in that the adapter includes means: for communicating with the client and the server and a configuration file in which the conversion service (or services) is (or are) declared.\",\n \"The adapter comprises to advantage a communication module able to manage communication with the client and/or a module for extracting headers and content from the Internet file and/or a module for pre-analysing headers and/or a module for creating interface files between the adapter and the server and/or a module for calling up the server's adaptation service and/or a response module and/or a module for checking the integrity of the converted file intended to the client and/or a module for restoring the converted file intended for the client.\",\n \"The adapter may be integrated with the server.\",\n \"The invention also relates to a method for exchanging flows between a client who has an Internet file, and a server able to provide at least one file conversion service, with communication between the client and the server being based on an iCAP Internet file content adaptation protocol, characterised in that it consists in placing an adapter of iCAP flows into files able to be used between the client and the server and in that it comprises the following steps: a—the sending of an Internet file, by said client to the adapter, b—the adaptation of the Internet file by the adapter and the sending of the adapted file to said server, c—the execution of the conversion of the adapted file, by said server and the sending of the converted file to said adapter, d—the adaptation of the converted file by the adapter e—the sending of the converted and adapted file to said client.\",\n \"According to one characteristic of the invention, step b—consists for the adapter in extracting the headers and possibly the content of the Internet file provided by the client, pre-analysing the headers, generating interface files, calling up the server's conversion service, awaiting a response from the server.\",\n \"Step d—consists to advantage for the adapter in verifying the call return parameter and checking the integrity of the converted file, when the integrity check requests it, restoring the converted file.\",\n \"According to an additional characteristic, the method according to the invention comprises a previous step of initialising the adapter consisting in declaring the conversion service or services in an adapter configuration file.\",\n \"Other particularities and advantages of the invention will emerge clearly from reading the description given by way of a non-restrictive example and in relation to the appended drawings in which: FIG.\",\n \"1 already described shows diagrammatically the steps of calling upon a file conversion system, FIG.\",\n \"2 already described shows diagrammatically the operation of a current file conversion system, FIG.\",\n \"3 shows diagrammatically a file conversion system according to the invention, FIG.\",\n \"4 shows diagrammatically the main operational elements of an iCAP flow exchange device according to the invention, FIG.\",\n \"5 shows diagrammatically an example of an Internet file, FIGS.\",\n \"6a), 6b), and 6c) show diagrammatically operational elements of a device according to the invention during conversion service call up, the execution of this service and the response respectively, FIG.\",\n \"7 is a diagram of relationships between classes.\",\n \"As shown in FIG.\",\n \"3, the invention consists in providing an adapter 5 preferably in the server 4, so as, Ion the one hand in the client-server direction, to adapt files so that they may be directly used by the file conversion services 40 and, on the other hand in the server-client direction, to provide a check on the integrity of the converted file and possibly its restoration.\",\n \"The adapter 5 makes it possible to formalise the exchanges between the iCAP client 3 and the file conversion services-by means of an adaptation of the iCAP flows in steps d) and e) in order to make them compatible with file conversion services 40 without it being necessary to develop an iCAP layer at file conversion service level.\",\n \"This then makes it possible to define file conversion services that are modular, open-ended and independent of the iCAP layers.\",\n \"The operation of the adapter according to the invention will now be described in relation to FIG.\",\n \"4: the adapter 5 is firstly initialised in order particularly to know the conversion service or services that may be called upon: in a configuration file are 1entered moreover operational parameters (communication port, size of the conversion service request waiting queue, directory where interface files with the server are generated, etc), the names of the conversion services and the specific parameters associated with the conversion services and the way to invoke them (access path, service compilation model (using static or dynamic libraries or executables), etc.\",\n \"); in one embodiment example, the operational parameters were initialised by the following fields and values: the communication port, server.default_port=8080 the size of the waiting queue, server.default_backlog=20 the directory, server.temp directory=/home/cbe/files the access path to specific files of a filter service, SVC.def_directory=/home/filtrage/SVC/def since it is initialised, the adapter 5 waits for the iCAP client 3 to connect via a communication module 50 thus allowing it to converse with the iCAP client 3 and to store temporarily all data exchanged during the communication phases.\",\n \"When file conversion involves a response from the Web server 2, this data shown in FIG.\",\n \"5 comprises, in addition to a content, the HTTP headers sent from the Web browser of the computer 1, the HTTP headers returned by the destination Web server, as well as the iCAP headers relating to the client 3.When file conversion involves a request, this data comprises the HTTP headers sent from the Web browser of the computer 1, as well as the iCAP headers relating to the iCAP client 3; the other data exists possibly in the event of the conversion being requested after step c) as shown in FIG.\",\n \"1, but is not involved since it is a request conversion.\",\n \"Reference is now made to FIG.\",\n \"4.The module 50 manages the TCP/IP connection and the iCAP protocol in a conventional way by opening particularly a TCP communication port; this module 50 is in wait mode so long as no iCAP request reaches it.\",\n \"When an iCAP request is received, the module sets up a new communication socket responsible for processing the conversion service requested by the client 3; the communication port so released returns to wait mode for another request.\",\n \"The following written instruction in C++ language in the LINUX operating system may for example be used: The iCAPAdapter>handleNewClients( ); The data sent by the client 3 is transmitted to a module 500 via the communication socket that has been created.\",\n \"The communication module 50 additionally comprises temporary storage means of all the data exchanged during the communication phases, by means of the module 500, the adapter 5 extracts the headers and the content from the data received from the module 50, analyses the headers, creates four files isolating the different headers from the content, using for example the instruction int iCr+CreateFiles(new_fd,cCanal,Exceed); after exact shaping the adapter 5 then redirects the data flow to the requested conversion service 40 using for example the instruction if(_ServiceIcap)returnValue-_ServiceIcap →CallService(new.fd,cCanal,_icapRootDirectory,Exceed); the service requested 40 may be localised on the server 4 or be remote and invoked through the Internet network for example.\",\n \"The module 500 then puts itself into wait mode for a response from the conversion service; the converted file returned by the service 40 is checked by the module 500 which verifies its integrity; the converted file will possibly be subject to shaping, or even to a set of additional actions to make it comply with the iCAP protocol; the checked file is delivered to the iCAP client 3 via the communication module 50 and the communication socket opened to handle this conversion service is closed, using for example the instruction CiCapGlobals::ManageResponse(Filtrage,Long_file,root Directory,socket_fd);.\",\n \"The diagram shown in FIG.\",\n \"7 illustrates the relationships existing between the classes used in the previous instructions.\",\n \"We will now describe in more detail the functionalities of the module 500 in relation to FIGS.\",\n \"6a), 6b), and 6c) corresponding to a conversion service call, conversion execution and response respectively.\",\n \"When a conversion service is called upon, the data is transmitted via the communication module 50 to a header and content extraction module 51.This module 51 separates the different fields specific to each of the headers and invokes the header pre-analysis module 52 for each header.\",\n \"The header pre-analysis module 52 verifies initially that the conversion service invoked is available on the iCAP server; in the event, it verifies that the conversion service invoked is described in the adapter's configuration file.\",\n \"If this is not the case, it returns an error to the iCAP client 3, via the communication module 50, without passing through the iCAP Server.\",\n \"The module 52 also analyses the iCAP request type: Web response modification or Web request modification.\",\n \"In request modification mode, it is not generally necessary to extract the content or even the HTTP header from the Web server.\",\n \"The module 52 identifies any parameters associated with the execution of the service.\",\n \"The module 52 returns the results to the extraction module 51 which calls on modules 53a, 53b, 53c, 53d to create output interface files.\",\n \"IThe modules 53a, 53b, 53c, 53d create the four files offering the file conversion services 40 a common interface, which facilitates the conversions to be carried out on the files: the header of the iCAP server 4 is entered into an iCAP-Out file which then contains the information relating to the iCAP client 3 as well as a number of HTTP request or response fields depending on whether it is request modification mode or response modification mode, the HTTP header sent by the computer 1 which contains information relating to the client browser as well as to the Web server invoked is entered into an HTTPClient-out file, the HTTP header returned by the Web server 2 which contains information relating to the content of the response, its URL address, its type, its length is entered into an HTTPServ-out file; this file may be empty in the case of request modification node; the content of the Web server response, expurgated of the headers (html file, gif, jpeg, etc), is entered into an F-Content-Out file; this file may be empty in the case of request modification mode.\",\n \"Each file name is completed by an index number incremented by one unit at each new iCAP request.\",\n \"Each file creation module 53a, 53b, 53c, 53d sends a return code to the extraction module 51 to indicate the result of the operation, as well as the names of the files.\",\n \"Once the headers are analysed and the files created, the extraction module 51 invokes the module 54 for calling up the conversion services 40.This module 54 calls up the relevant conversion service by providing a set of parameters comprising particularly the name of the service call function, the address and the iCAP server port where the service is located and the names of the previously defined interface files.\",\n \"As shown in FIG.\",\n \"6b), the conversion service 40 then executes the conversion in relation to modules 53a, 53b, 53c, 53d allowing the interface files to be accessed, and returns the converted data to a response module 55.The response module 55 shown in FIG.\",\n \"6c), recovers the data from the conversion service 40.Among this data there features a service call function return code indicating that the conversion has been correctly carried out; when this is not the case and a problem has been detected, the return code, which is then an extended error code, allows the response module 55 to make a fast diagnosis of the problem detected.\",\n \"Among this data there also features additional information in the form of four input interface files corresponding to the output interface files.\",\n \"Each of these files reflects the file conversion that has occurred; the result is: an additional modification of the iCAP header, possible modifications of the client HTTP header, possible modifications of the server HTTP header, the data resulting from the converted file.\",\n \"The response module 55 may use these files to call up a new iCAP service and thus link the conversion services together in so far as the data and parameters are available and already listed in the directory.\",\n \"This functionality may be activated when the services to be linked together have the same generic name.\",\n \"A virus checking service may for example follow a “parental” control service preventing access to Web pages for which licentious topics have been detected, these two services being recognised under the generic name “control”.\",\n \"After receiving the converted data, the response module 55 invokes the integrity check module 56.The module 56 verifies the integrity of the data by carrying out particularly coherence tests between for example the length of the converted file and the header information relating to this length.\",\n \"The checked data is then sent to the file restoration module 57.The module 57 creates or completes the iCAP header with the information on the nature of the iCAP server 4.It modifies the HTTP headers of the computer 1 and/or of the Web server 2 as a function of the data in the integrity check module 56; in the event of the absence of one or more input interface files, the module 57 may recreate them from the output interface files.\",\n \"Once restored, the file is transmitted to the communication module 50, which returns it to the iCAP client 3 who has initialised the iCAP request.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473984"}}},{"rowIdx":777,"cells":{"text":{"kind":"list like","value":[["Method for eliminating metal halides that are present in a liquid or gaseous, organic or non-organic effluent","The invention relates to a method for eliminating metal halides which are present in a liquid or gaseous, organic or non-organic effluent.","According to the invention, the elimination is carried out by absorption of said metal halides on alumina agglomerates.","The inventive method is characterised in that: the specific surface area of said agglomerates is between 50 and 350 m2/g, preferably between 70 and 300 m2/g and, better still, between 80 and 250 m2/g; and the V80Å thereof is greater than or equal to 20 ml/100 g, preferably greater than or equal to 25 ml/100 g, better still greater than or equal to 30 ml/100 g and, optimally, greater than or equal to 35 ml/100 g."],["1.A method of eliminating metal halides that are present in a liquid or gaseous, organic or non-organic effluent, in which this elimination is carried out by adsorption of said metal halides on alumina agglomerates, characterized in that wherein: said agglomerates have a specific surface area of between 50 and 350 m2/g, preferably between 70 and 300 m2/g and even more preferably between 80 and 250 m2/g; and wherein said agglomerates have a V80Å of greater than or equal to 20 ml/100 g, preferably greater than or equal to 25 ml/100 g, even more preferably greater than or equal to 30 ml/100 g and optimally greater than or equal to 35 ml/100 g. 2.The method as claimed in claim 1, wherein said agglomerates have a V400Å of greater than or equal to 10 ml/100 g, preferably greater than or equal to 15 ml/100 g and even more preferably greater than or equal to 20 ml/100 g. 3.The method as claimed in claim 1 wherein said agglomerates have a V37Å of greater than or equal to 45 ml/100 g and preferably greater than or equal to 55 ml/100 g. 4.The method as claimed in claim 1 wherein said agglomerates comprise one or more dopant compounds selected from compounds of alkali metals, alkaline-earth metals and rare earths, having a maximum content of 20%, preferably less than 10%.","5.The method as claimed in claim 1 wherein said agglomerates are in the form of beads.","6.The method as claimed in claim 5, wherein said beads have a diameter of less than or equal to 8 mm, preferably between 1 and 5 mm.","7.The method as claimed in claim 1 wherein said agglomerates are in the form of extruded materials.","8.The method as claimed in claim 7, wherein said extruded materials are of cylindrical shape.","9.The method as claimed in claim 7, wherein said extruded materials are of polylobate shape.","10.The method as claimed in claim 7 wherein said extruded materials have an inscribed diameter of their cross section of less than or equal to 4 mm.","11.The method as claimed in claim 1 wherein said alumina agglomerates are in the form of powder.","12.The method as claimed in claim 1 wherein-said effluent is a dichloroethylene-based medium and in that said metal halide is ferric chloride."],["The invention relates to the field of the elimination of impurities contained in organic or non-organic industrial effluents in the liquid or gaseous state.","More precisely, it relates to the elimination of impurities consisting of metal halides contained in these effluents by absorption on alumina agglomerates.","Many gaseous or liquid industrial effluents contain impurities that it is desirable to eliminate.","These impurities may pose problems of various types.","Among these mention may be made of: interference with processes in which the effluent participates, for example formation of undesirable products or inhibition or poisoning of a catalyst; degradation of the quality of the final product resulting from a lack of purity, from undesirable coloration, etc.","; and formation of industrial waste coming from treatment of the effluent, said waste being difficult to reprocess and therefore posing environmental problems.","It is known to improve the purity of certain liquid or gaseous, organic or non-organic industrial effluents by passing them over a mineral material, such as alumina, that retains certain impurities by adsorption on its surface.","In particular, it is known to use alumina agglomerates to purify industrial effluents containing metal halides in trace quantities that it is desired to eliminate.","It is usually accepted that these alumina particles must have a high specific surface area and therefore predominantly pores of very small size.","The object of the invention is to propose a method of purifying industrial effluents containing metal halides by selective adsorption on alumina, using forms of alumina that are relatively inexpensive to manufacture but nevertheless are extremely effective for the application envisioned, much more effective than the alumina particles for this purpose in the prior art.","For this purpose, the subject of the invention is a method of eliminating metal halides that are present in a liquid or gaseous, organic or non-organic effluent, in which this elimination is carried out by adsorption of said metal halides on alumina agglomerates, characterized in that: said agglomerates have a specific surface area of between 50 and 350 m2/g, preferably between 70 and 300 m2/g and even more preferably between 80 and 250 m2/g; and in that said agglomerates have a V80Å of greater than or equal to 20 ml/100 g, preferably greater than or equal to 25 ml/100 g, even more preferably greater than or equal to 30 ml/100 g and optimally greater than or equal to 35 ml/100 g. Preferably, said agglomerates have a V400Å of greater than or equal to 10 ml/100 g, preferably greater than or equal to 15 ml/100 g and even more preferably greater than or equal to 20 ml/100 g. Preferably, said agglomerates have a V37Å of greater than or equal to 45 ml/100 g and preferably greater than or equal to 55 ml/100 g. Said agglomerates may include one or more dopant compounds selected from compounds of alkali metals, alkaline-earth metals and rare earths, having a maximum content of 20%, preferably less than 10%.","Said agglomerates may be in the form of beads, preferably having a diameter of less than or equal to 8 mm and more preferably between 1 and 5 mm.","Said agglomerates may be in the form of extruded materials, for example with a cylindrical or polylobate shape.","Said extruded materials preferably have an inscribed diameter of their cross section of less than or equal to 4 mm.","Said alumina agglomerates may be in the form of powder.","In one particular application of the invention, said effluent is found downstream of a polyvinyl chloride production unit.","The medium may then be based on dichloroethylene and said metal halide is ferric chloride.","As will have been understood, the invention consists in using, as selective adsorbent materials, a class of alumina agglomerates exhibiting particular characteristics in terms of both specific surface area and of porous structure.","Surprisingly, in considering what had previously been determined experimentally, these agglomerates must have a relatively low specific surface area and a porosity profile in which pores of very small diameter do not necessarily represent a very large volume.","However, these agglomerates possess remarkably high adsorbent properties with respect to metal halides contained in liquid or gaseous, organic or non-organic industrial effluents (it being possible, for example, for the latter to be aqueous solutions).","The invention will be more clearly understood from the description that follows, given with reference to the single appended figure.","This shows, for various alumina agglomerates according to the invention and various control alumina agglomerates, the degree of elimination (in percent) of the ferric chloride contained in a ferric chloride solution in acetophenone brought into contact with an alumina agglomerate after 37 hours of reaction.","A preferential, but in no way limiting, application of the invention is downstream of a polyvinyl chloride (PVC) production line.","After polymerization of the PVC, a residual dichloroethylene-based medium may remain which contains traces of ferric chloride.","This ferric chloride must be eliminated before such a residual medium is recycled.","It is also known to carry out this elimination by adsorption on alumina agglomerates such as those mentioned later with regard to the reference materials.","It will be seen that the use, for this purpose, of one particular class of alumina agglomerates substantially improves the efficiency of a ferric chloride elimination operation.","The alumina agglomerates used within the context of the method of eliminating metal halides according to the invention must necessarily have a specific surface area of between 50 and 350 m2/g, preferably between 70 and 300 m2/g and advantageously between 80 and 250 m2/g.","Also necessarily, they have a volume occupied by pores with a diameter of greater than or equal to 80 Å (denoted in short by the notation V80Å) of greater than or equal to 20 ml/100 g, preferably greater than or equal to 25 ml/100 g, advantageously greater than or equal to 30 ml/100 g or even than greater than or equal to 35 ml/100 g. It will be noted that the presence of pores having a diameter of less than 80 Å is of only little importance within the context of the invention.","The little importance ascribed to such microporosity goes, as mentioned, counter to what has been commonly accepted hitherto.","According to a preferred variant of the invention, the volume occupied by the pores having a diameter of greater than or equal to 400 Å (V400Å) is greater than or equal to 10 ml/100 g, advantageously greater than or equal to 20 ml/100 g. The V80Å and V400Å may be determined by a conventional mercury porosimetry method.","For this purpose, the alumina specimen is first placed in a column into which mercury under a pressure P is introduced.","Since mercury does not wet alumina, its penetration or non-penetration into the pores of the specimen having a given diameter depends on the value of P. In order to be filled, the finest pores require a higher pressure P to be established than for filling the coarser pores.","By measuring the amount of mercury penetrating the specimen for various values of P, it is possible to determine the volume occupied by the pores of diameter greater than given values of this diameter.","According to one particular form of the invention, the alumina agglomerates may be chemically modified by the addition of alkali metal or alkaline-earth metal compounds, or rare-earth compounds, or a mixture of such compounds.","Preferably, compounds based on sodium, potassium, calcium, magnesium or lanthanum are chosen.","Sodium is a preferred example, and may be introduced in the form of one or more precursors of its oxide Na2O.","The addition of one or more dopant compounds may be carried out before or after the forming operation or during it.","The dopant compounds are present in the alumina agglomerate with a total mass content of less than 20%, preferably less than 10%.","These dopant compounds enhance the adsorbent properties of the surface of the alumina agglomerates with respect to the metal halide molecules that it is desired to eliminate.","The alumina may be used in powder form, but preferably it is used after a forming step.","Beads, advantageously with a diameter of less than 8 mm, preferably mostly between 1 and 5 mm, constitute a preferred form of the alumina agglomerates according to the invention.","Another preferred form is that of cylindrical or polylobate extrudates, the inscribed diameter of their cross section preferably being less than 4 mm.","The beads may be obtained by means of a rotational technique, by agglomeration of an alumina powder in a pelletizer or drum.","This type of process makes it possible to obtain, in a known manner, beads with controlled pore distributions and diameters, these distributions and dimensions being, in general, created during the agglomeration step.","The porosity may be created by various means, such as the choice of the particle size of the alumina powder or the agglomeration of several alumina powders of different particle sizes.","Another method consists in mixing with the alumina powder, before or during the agglomeration step, a compound, called a pore former, that disappears when it is heated and thus creates porosity in the beads.","As pore-former compounds used, mention may be made, by way of example, of wood flour, charcoal, sulfur, tars, plastics or emulsions of plastics, such as polyvinyl chloride and polyvinyl alcohols, naphthalene or the like.","The amount of pore-former compounds added is determined by the desired volume.","One or more heat treatments then complete the operation of forming the beads.","The extrudates may be obtained by mixing and then extruding an alumina gel or an alumina powder or a mixture of various raw materials.","The initial alumina powder may be obtained conventionally by rapid dehydration of an aluminium hydroxide (for example, hydrargilite).","The addition of one or more dopant compounds may be carried out before or after the forming operation or during the latter.","As examples, the ferric chloride (FeCl3) adsorption results for various control alumina agglomerates and for alumina agglomerates having the characteristics required by the method according to the invention will be compared.","Eight alumina agglomerates were considered, the use of aluminas A, B and C forming a part of the prior art and the use of aluminas D, E, F, G and H corresponding to the invention.","100 ppm of FeCl3.6H2O were placed in a beaker containing 250 ml of acetophenone.","Next, 1 g of alumina pretreated at 300° C., in the form of beads or extruded materials, was then added.","The beaker was then isolated from the ambient air, shielded from light in order to avoid any degradation of the solvent (which was photosensitive) and subjected to magnetic stirring, the beads or extrudates being isolated from the bar magnet so as to avoid any undesirable attrition during the experiment.","The description of the alumina agglomerates used is given in table I, in which the diameters are in mm, the specific surface areas are in m2/g and the porosities are in ml/100 g. Table I mentions, apart from the abovementioned parameters, the V37Å of each agglomerate, that is to say the volume occupied by the pores having a diameter greater than or equal to 37 Å.","The difference between V37Å and V80Å is representative of the amount of pores with very small diameters of the agglomerate tested.","Finally, the Na2O content of the agglomerates, expressed in ppm, is mentioned.","TABLE I Characteristics of the alumina agglomerates tested Controls Invention Alumina A B C D E F G H Form Beads Beads Beads Extrudates Beads Beads Extrudates Beads Diameter 1.4-2.8 1.4-2.8 2-4 1.2 2.0-2.8 1.4-2.8 1.2 2.0-2.8 Specific 337 257 6 266 196 150 251 172 surface area V37Å 35.6 37.2 53.5 68.4 68.0 102.3 59.8 58.4 V80Å 14.1 9.4 53.4 48.9 57.5 98.5 41.5 46.5 V400Å 5.3 4.6 53.2 5.8 20.0 53.7 5.4 15.2 Na2O 3500 20000 700 500 700 700 20000 20000 The iron chloride concentration of the acetophenone solution was monitored by UV-visible analysis, in particular by monitoring the change in absorbance at the wavelength of 378 nm.","After 37 hours of reaction at room temperature, the degrees of elimination of iron chloride from the organic solution were thus determined, and these are plotted in the diagram shown in FIG.","1.The aluminas used in the prior art show an adsorption potential markedly inferior to that of the aluminas used in the method according to the invention.","The ferric chloride adsorption efficiency of control alumina C, having only large-diameter pores and a very low specific surface area, is very mediocre.","It should be noted that the aluminas of the method according to the invention have a specific surface area that is not particularly high: it is of the same order of magnitude or substantially less than that of control alumina B and substantially less than that of control alumina A.","Compared with control aluminas A and B, that have a similar or higher specific surface area, the aluminas according to the invention are distinguished by their relatively high V80Å.","However, it should also be noted that, in many cases, the aluminas according to the invention exhibit a small difference between V80Å and V37Å, this being an indicator of the fact that they have few pores of very small size.","In particular, this is the case for alumina F that gives the best ferric chloride adsorption results.","However, it will be preferable for V37Å to be usefully at least 45 ml/100 g, preferably greater than 55 ml/100 g. It should also be noted that this alumina F has a high V400Å, and therefore large-diameter pores present at a relatively high quantity.","This all results in quite a low specific surface area, which nevertheless does not compromise the quality of the results obtained, quite to the contrary.","These results therefore go counter to what was commonly accepted as having to constitute preferred characteristics of the alumina agglomerates in their envisaged application of adsorbing metal chlorides.","Finally, it should be noted that the aluminas of the examples according to the invention do not have very high Na2O (dopant compound) contents; they are at most 2%.","Despite this, they exhibit ferric chloride adsorption properties substantially higher than the control aluminas of comparable Na2O content.","This clearly shows that, for this application, the role of the porosity of the agglomerates is predominant, the doping of these agglomerates by alkali or alkaline-earth metal compounds being merely a variant of the invention.","The invention is not limited to the specific examples that have been mentioned, namely the adsorption of ferric chloride from an effluent based on dichloroethylene or acetophenone.","The use of alumina agglomerates as described for the adsorption of metal halides may be envisioned for the treatment of any gaseous or liquid, organic or non-organic effluents.","It is particularly applicable to aqueous solutions."]],"string":"[\n [\n \"Method for eliminating metal halides that are present in a liquid or gaseous, organic or non-organic effluent\",\n \"The invention relates to a method for eliminating metal halides which are present in a liquid or gaseous, organic or non-organic effluent.\",\n \"According to the invention, the elimination is carried out by absorption of said metal halides on alumina agglomerates.\",\n \"The inventive method is characterised in that: the specific surface area of said agglomerates is between 50 and 350 m2/g, preferably between 70 and 300 m2/g and, better still, between 80 and 250 m2/g; and the V80Å thereof is greater than or equal to 20 ml/100 g, preferably greater than or equal to 25 ml/100 g, better still greater than or equal to 30 ml/100 g and, optimally, greater than or equal to 35 ml/100 g.\"\n ],\n [\n \"1.A method of eliminating metal halides that are present in a liquid or gaseous, organic or non-organic effluent, in which this elimination is carried out by adsorption of said metal halides on alumina agglomerates, characterized in that wherein: said agglomerates have a specific surface area of between 50 and 350 m2/g, preferably between 70 and 300 m2/g and even more preferably between 80 and 250 m2/g; and wherein said agglomerates have a V80Å of greater than or equal to 20 ml/100 g, preferably greater than or equal to 25 ml/100 g, even more preferably greater than or equal to 30 ml/100 g and optimally greater than or equal to 35 ml/100 g. 2.The method as claimed in claim 1, wherein said agglomerates have a V400Å of greater than or equal to 10 ml/100 g, preferably greater than or equal to 15 ml/100 g and even more preferably greater than or equal to 20 ml/100 g. 3.The method as claimed in claim 1 wherein said agglomerates have a V37Å of greater than or equal to 45 ml/100 g and preferably greater than or equal to 55 ml/100 g. 4.The method as claimed in claim 1 wherein said agglomerates comprise one or more dopant compounds selected from compounds of alkali metals, alkaline-earth metals and rare earths, having a maximum content of 20%, preferably less than 10%.\",\n \"5.The method as claimed in claim 1 wherein said agglomerates are in the form of beads.\",\n \"6.The method as claimed in claim 5, wherein said beads have a diameter of less than or equal to 8 mm, preferably between 1 and 5 mm.\",\n \"7.The method as claimed in claim 1 wherein said agglomerates are in the form of extruded materials.\",\n \"8.The method as claimed in claim 7, wherein said extruded materials are of cylindrical shape.\",\n \"9.The method as claimed in claim 7, wherein said extruded materials are of polylobate shape.\",\n \"10.The method as claimed in claim 7 wherein said extruded materials have an inscribed diameter of their cross section of less than or equal to 4 mm.\",\n \"11.The method as claimed in claim 1 wherein said alumina agglomerates are in the form of powder.\",\n \"12.The method as claimed in claim 1 wherein-said effluent is a dichloroethylene-based medium and in that said metal halide is ferric chloride.\"\n ],\n [\n \"The invention relates to the field of the elimination of impurities contained in organic or non-organic industrial effluents in the liquid or gaseous state.\",\n \"More precisely, it relates to the elimination of impurities consisting of metal halides contained in these effluents by absorption on alumina agglomerates.\",\n \"Many gaseous or liquid industrial effluents contain impurities that it is desirable to eliminate.\",\n \"These impurities may pose problems of various types.\",\n \"Among these mention may be made of: interference with processes in which the effluent participates, for example formation of undesirable products or inhibition or poisoning of a catalyst; degradation of the quality of the final product resulting from a lack of purity, from undesirable coloration, etc.\",\n \"; and formation of industrial waste coming from treatment of the effluent, said waste being difficult to reprocess and therefore posing environmental problems.\",\n \"It is known to improve the purity of certain liquid or gaseous, organic or non-organic industrial effluents by passing them over a mineral material, such as alumina, that retains certain impurities by adsorption on its surface.\",\n \"In particular, it is known to use alumina agglomerates to purify industrial effluents containing metal halides in trace quantities that it is desired to eliminate.\",\n \"It is usually accepted that these alumina particles must have a high specific surface area and therefore predominantly pores of very small size.\",\n \"The object of the invention is to propose a method of purifying industrial effluents containing metal halides by selective adsorption on alumina, using forms of alumina that are relatively inexpensive to manufacture but nevertheless are extremely effective for the application envisioned, much more effective than the alumina particles for this purpose in the prior art.\",\n \"For this purpose, the subject of the invention is a method of eliminating metal halides that are present in a liquid or gaseous, organic or non-organic effluent, in which this elimination is carried out by adsorption of said metal halides on alumina agglomerates, characterized in that: said agglomerates have a specific surface area of between 50 and 350 m2/g, preferably between 70 and 300 m2/g and even more preferably between 80 and 250 m2/g; and in that said agglomerates have a V80Å of greater than or equal to 20 ml/100 g, preferably greater than or equal to 25 ml/100 g, even more preferably greater than or equal to 30 ml/100 g and optimally greater than or equal to 35 ml/100 g. Preferably, said agglomerates have a V400Å of greater than or equal to 10 ml/100 g, preferably greater than or equal to 15 ml/100 g and even more preferably greater than or equal to 20 ml/100 g. Preferably, said agglomerates have a V37Å of greater than or equal to 45 ml/100 g and preferably greater than or equal to 55 ml/100 g. Said agglomerates may include one or more dopant compounds selected from compounds of alkali metals, alkaline-earth metals and rare earths, having a maximum content of 20%, preferably less than 10%.\",\n \"Said agglomerates may be in the form of beads, preferably having a diameter of less than or equal to 8 mm and more preferably between 1 and 5 mm.\",\n \"Said agglomerates may be in the form of extruded materials, for example with a cylindrical or polylobate shape.\",\n \"Said extruded materials preferably have an inscribed diameter of their cross section of less than or equal to 4 mm.\",\n \"Said alumina agglomerates may be in the form of powder.\",\n \"In one particular application of the invention, said effluent is found downstream of a polyvinyl chloride production unit.\",\n \"The medium may then be based on dichloroethylene and said metal halide is ferric chloride.\",\n \"As will have been understood, the invention consists in using, as selective adsorbent materials, a class of alumina agglomerates exhibiting particular characteristics in terms of both specific surface area and of porous structure.\",\n \"Surprisingly, in considering what had previously been determined experimentally, these agglomerates must have a relatively low specific surface area and a porosity profile in which pores of very small diameter do not necessarily represent a very large volume.\",\n \"However, these agglomerates possess remarkably high adsorbent properties with respect to metal halides contained in liquid or gaseous, organic or non-organic industrial effluents (it being possible, for example, for the latter to be aqueous solutions).\",\n \"The invention will be more clearly understood from the description that follows, given with reference to the single appended figure.\",\n \"This shows, for various alumina agglomerates according to the invention and various control alumina agglomerates, the degree of elimination (in percent) of the ferric chloride contained in a ferric chloride solution in acetophenone brought into contact with an alumina agglomerate after 37 hours of reaction.\",\n \"A preferential, but in no way limiting, application of the invention is downstream of a polyvinyl chloride (PVC) production line.\",\n \"After polymerization of the PVC, a residual dichloroethylene-based medium may remain which contains traces of ferric chloride.\",\n \"This ferric chloride must be eliminated before such a residual medium is recycled.\",\n \"It is also known to carry out this elimination by adsorption on alumina agglomerates such as those mentioned later with regard to the reference materials.\",\n \"It will be seen that the use, for this purpose, of one particular class of alumina agglomerates substantially improves the efficiency of a ferric chloride elimination operation.\",\n \"The alumina agglomerates used within the context of the method of eliminating metal halides according to the invention must necessarily have a specific surface area of between 50 and 350 m2/g, preferably between 70 and 300 m2/g and advantageously between 80 and 250 m2/g.\",\n \"Also necessarily, they have a volume occupied by pores with a diameter of greater than or equal to 80 Å (denoted in short by the notation V80Å) of greater than or equal to 20 ml/100 g, preferably greater than or equal to 25 ml/100 g, advantageously greater than or equal to 30 ml/100 g or even than greater than or equal to 35 ml/100 g. It will be noted that the presence of pores having a diameter of less than 80 Å is of only little importance within the context of the invention.\",\n \"The little importance ascribed to such microporosity goes, as mentioned, counter to what has been commonly accepted hitherto.\",\n \"According to a preferred variant of the invention, the volume occupied by the pores having a diameter of greater than or equal to 400 Å (V400Å) is greater than or equal to 10 ml/100 g, advantageously greater than or equal to 20 ml/100 g. The V80Å and V400Å may be determined by a conventional mercury porosimetry method.\",\n \"For this purpose, the alumina specimen is first placed in a column into which mercury under a pressure P is introduced.\",\n \"Since mercury does not wet alumina, its penetration or non-penetration into the pores of the specimen having a given diameter depends on the value of P. In order to be filled, the finest pores require a higher pressure P to be established than for filling the coarser pores.\",\n \"By measuring the amount of mercury penetrating the specimen for various values of P, it is possible to determine the volume occupied by the pores of diameter greater than given values of this diameter.\",\n \"According to one particular form of the invention, the alumina agglomerates may be chemically modified by the addition of alkali metal or alkaline-earth metal compounds, or rare-earth compounds, or a mixture of such compounds.\",\n \"Preferably, compounds based on sodium, potassium, calcium, magnesium or lanthanum are chosen.\",\n \"Sodium is a preferred example, and may be introduced in the form of one or more precursors of its oxide Na2O.\",\n \"The addition of one or more dopant compounds may be carried out before or after the forming operation or during it.\",\n \"The dopant compounds are present in the alumina agglomerate with a total mass content of less than 20%, preferably less than 10%.\",\n \"These dopant compounds enhance the adsorbent properties of the surface of the alumina agglomerates with respect to the metal halide molecules that it is desired to eliminate.\",\n \"The alumina may be used in powder form, but preferably it is used after a forming step.\",\n \"Beads, advantageously with a diameter of less than 8 mm, preferably mostly between 1 and 5 mm, constitute a preferred form of the alumina agglomerates according to the invention.\",\n \"Another preferred form is that of cylindrical or polylobate extrudates, the inscribed diameter of their cross section preferably being less than 4 mm.\",\n \"The beads may be obtained by means of a rotational technique, by agglomeration of an alumina powder in a pelletizer or drum.\",\n \"This type of process makes it possible to obtain, in a known manner, beads with controlled pore distributions and diameters, these distributions and dimensions being, in general, created during the agglomeration step.\",\n \"The porosity may be created by various means, such as the choice of the particle size of the alumina powder or the agglomeration of several alumina powders of different particle sizes.\",\n \"Another method consists in mixing with the alumina powder, before or during the agglomeration step, a compound, called a pore former, that disappears when it is heated and thus creates porosity in the beads.\",\n \"As pore-former compounds used, mention may be made, by way of example, of wood flour, charcoal, sulfur, tars, plastics or emulsions of plastics, such as polyvinyl chloride and polyvinyl alcohols, naphthalene or the like.\",\n \"The amount of pore-former compounds added is determined by the desired volume.\",\n \"One or more heat treatments then complete the operation of forming the beads.\",\n \"The extrudates may be obtained by mixing and then extruding an alumina gel or an alumina powder or a mixture of various raw materials.\",\n \"The initial alumina powder may be obtained conventionally by rapid dehydration of an aluminium hydroxide (for example, hydrargilite).\",\n \"The addition of one or more dopant compounds may be carried out before or after the forming operation or during the latter.\",\n \"As examples, the ferric chloride (FeCl3) adsorption results for various control alumina agglomerates and for alumina agglomerates having the characteristics required by the method according to the invention will be compared.\",\n \"Eight alumina agglomerates were considered, the use of aluminas A, B and C forming a part of the prior art and the use of aluminas D, E, F, G and H corresponding to the invention.\",\n \"100 ppm of FeCl3.6H2O were placed in a beaker containing 250 ml of acetophenone.\",\n \"Next, 1 g of alumina pretreated at 300° C., in the form of beads or extruded materials, was then added.\",\n \"The beaker was then isolated from the ambient air, shielded from light in order to avoid any degradation of the solvent (which was photosensitive) and subjected to magnetic stirring, the beads or extrudates being isolated from the bar magnet so as to avoid any undesirable attrition during the experiment.\",\n \"The description of the alumina agglomerates used is given in table I, in which the diameters are in mm, the specific surface areas are in m2/g and the porosities are in ml/100 g. Table I mentions, apart from the abovementioned parameters, the V37Å of each agglomerate, that is to say the volume occupied by the pores having a diameter greater than or equal to 37 Å.\",\n \"The difference between V37Å and V80Å is representative of the amount of pores with very small diameters of the agglomerate tested.\",\n \"Finally, the Na2O content of the agglomerates, expressed in ppm, is mentioned.\",\n \"TABLE I Characteristics of the alumina agglomerates tested Controls Invention Alumina A B C D E F G H Form Beads Beads Beads Extrudates Beads Beads Extrudates Beads Diameter 1.4-2.8 1.4-2.8 2-4 1.2 2.0-2.8 1.4-2.8 1.2 2.0-2.8 Specific 337 257 6 266 196 150 251 172 surface area V37Å 35.6 37.2 53.5 68.4 68.0 102.3 59.8 58.4 V80Å 14.1 9.4 53.4 48.9 57.5 98.5 41.5 46.5 V400Å 5.3 4.6 53.2 5.8 20.0 53.7 5.4 15.2 Na2O 3500 20000 700 500 700 700 20000 20000 The iron chloride concentration of the acetophenone solution was monitored by UV-visible analysis, in particular by monitoring the change in absorbance at the wavelength of 378 nm.\",\n \"After 37 hours of reaction at room temperature, the degrees of elimination of iron chloride from the organic solution were thus determined, and these are plotted in the diagram shown in FIG.\",\n \"1.The aluminas used in the prior art show an adsorption potential markedly inferior to that of the aluminas used in the method according to the invention.\",\n \"The ferric chloride adsorption efficiency of control alumina C, having only large-diameter pores and a very low specific surface area, is very mediocre.\",\n \"It should be noted that the aluminas of the method according to the invention have a specific surface area that is not particularly high: it is of the same order of magnitude or substantially less than that of control alumina B and substantially less than that of control alumina A.\",\n \"Compared with control aluminas A and B, that have a similar or higher specific surface area, the aluminas according to the invention are distinguished by their relatively high V80Å.\",\n \"However, it should also be noted that, in many cases, the aluminas according to the invention exhibit a small difference between V80Å and V37Å, this being an indicator of the fact that they have few pores of very small size.\",\n \"In particular, this is the case for alumina F that gives the best ferric chloride adsorption results.\",\n \"However, it will be preferable for V37Å to be usefully at least 45 ml/100 g, preferably greater than 55 ml/100 g. It should also be noted that this alumina F has a high V400Å, and therefore large-diameter pores present at a relatively high quantity.\",\n \"This all results in quite a low specific surface area, which nevertheless does not compromise the quality of the results obtained, quite to the contrary.\",\n \"These results therefore go counter to what was commonly accepted as having to constitute preferred characteristics of the alumina agglomerates in their envisaged application of adsorbing metal chlorides.\",\n \"Finally, it should be noted that the aluminas of the examples according to the invention do not have very high Na2O (dopant compound) contents; they are at most 2%.\",\n \"Despite this, they exhibit ferric chloride adsorption properties substantially higher than the control aluminas of comparable Na2O content.\",\n \"This clearly shows that, for this application, the role of the porosity of the agglomerates is predominant, the doping of these agglomerates by alkali or alkaline-earth metal compounds being merely a variant of the invention.\",\n \"The invention is not limited to the specific examples that have been mentioned, namely the adsorption of ferric chloride from an effluent based on dichloroethylene or acetophenone.\",\n \"The use of alumina agglomerates as described for the adsorption of metal halides may be envisioned for the treatment of any gaseous or liquid, organic or non-organic effluents.\",\n \"It is particularly applicable to aqueous solutions.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473987"}}},{"rowIdx":778,"cells":{"text":{"kind":"list like","value":[["Semi-continuous photochemical method and device therefor","In the photochemical synthetic process in semi-continuous mode according to the invention, a reactor comprising two zones is used, the radiating portion of the lamp(s) being totally immersed in a first zone which is completely filled with reaction medium and spills off via an overflow into a second zone whose volume is sufficient to contain the volume of reaction medium originating from the first zone and corresponding substantially to the volume of the reagent(s) gradually introduced."],["1.Photochemical synthetic process in semi-continuous mode in which at least one of the reagents is introduced, from the start of the reaction, in total into the photochemical reactor, the other reagent(s) then being introduced gradually, characterized in that the process is performed in a reactor comprising two zones, the radiating portion of the lamp(s) being totally immersed in a first zone which is completely filled with reaction medium and spills off via an overflow into a second zone whose volume is sufficient to contain the volume of reaction medium originating from the first zone and corresponding substantially to the volume of the reagent(s) gradually introduced.","2.Process according to claim 1, in which the reagent initially introduced in total into the first zone is a mercaptan or hydrogen sulphide, the reagent gradually introduced being an alkene.","3.Process according to claim 1, in which a photoinitiator is also introduced gradually.","4.Process according to claim 1, in which one or more lamp(s) delivering UV radiation of wavelengths between 200 and 400 nm is(are) used.","5.Process according to one of claim 1, which is performed at a pressure ranging from 0.1 to 50 bar relative.","6.Process according to one of claim 1, which is performed at a temperature of between −20 and 120° C. 7.Process according to claim 1, which is performed at a temperature of between 10 and 90° C. 8.Process according to claim 2, in which the mercaptan is methyl mercaptan and the alkene is ethylene.","9.Process according to claim 1, in which the ratio of the volume of the first zone to the volume of the second zone is between 0.4 and 0.7.10.Process according to claim 5, which is performed at a pressure of between 2 and 10 bar relative.","11.Device for carrying out the process according to claim 1, characterized in that it essentially comprises: a first zone 1 in which at least one lamp 2 is placed such that at least all of its radiating portion 2a is totally immersed, a second zone 3 adjacent to the zone 1 and serving as an overspill therefor, pipes 4 and 5 and a distributor 9 for ensuring, via at least one pump 6 and a heat exchanger 7, circulation of the reaction medium from the zone 3 to the zone 1, at least one pipe 8 for introducing the reagents, a degassing vent 10, and a discharge pipe 11."],["<SOH> BACKGROUND OF THE INVENTION <EOH>The expression “process performed in semi-continuous mode” means here a process in which at least one of the reagents is, from the start of the reaction, introduced in total into the photochemical reactor while at least one of the reagents is introduced gradually into the reactor as it is consumed.","Such a procedure is occasionally found to be necessary in order to observe the optimum synthetic conditions.","In a photochemical process operating in liquid medium, it is necessary for the radiating portion of the lamp to be constantly fully immersed in the liquid reaction medium.","This total immersion is essential so as not to illuminate a gas phase whose behaviour, under ultraviolet irradiation, may prove to be hazardous or an inconvenience for the reaction under consideration.","Thus, for example, in the synthesis of sulphides and mercaptans by photochemical reaction of an alkene with a mercaptan or hydrogen sulphide, UV irradiation of the gas phase may produce elemental sulphur which is a well-known inhibitor of free-radical reactions such as those carried out to produce sulphides or mercaptans.","Moreover, total immersion of the radiating portion of the lamp also makes it possible to remove, via the reaction medium, the heat emitted by the lamp; this arrangement simplifies the process and minimizes the equipment required (jacket, cooling-water circuit, exchangers, etc.).","However, this arrangement is incompatible with the use of a semi-continuous process in which the gradual introduction of at least one of the reagents entails an increase in the volume of the reaction medium in a reactor of fixed geometry."],["<SOH> SUMMARY OF THE INVENTION <EOH>According to the invention, this problem has been solved by carrying out the photochemical process in a reactor comprising two zones, the radiating portion of the lamp being totally immersed in a first zone which is completely filled with reaction medium and spills off via an overflow into a second zone whose volume is sufficient to contain the volume of reaction medium originating from the first zone and corresponding substantially to the volume of the reagent(s) gradually introduced."],["FIELD OF THE INVENTION The present invention relates to the field of photochemical processes and its subject is, more particularly, a photochemical process performed semi-continuously, and also a device for carrying it out.","BACKGROUND OF THE INVENTION The expression “process performed in semi-continuous mode” means here a process in which at least one of the reagents is, from the start of the reaction, introduced in total into the photochemical reactor while at least one of the reagents is introduced gradually into the reactor as it is consumed.","Such a procedure is occasionally found to be necessary in order to observe the optimum synthetic conditions.","In a photochemical process operating in liquid medium, it is necessary for the radiating portion of the lamp to be constantly fully immersed in the liquid reaction medium.","This total immersion is essential so as not to illuminate a gas phase whose behaviour, under ultraviolet irradiation, may prove to be hazardous or an inconvenience for the reaction under consideration.","Thus, for example, in the synthesis of sulphides and mercaptans by photochemical reaction of an alkene with a mercaptan or hydrogen sulphide, UV irradiation of the gas phase may produce elemental sulphur which is a well-known inhibitor of free-radical reactions such as those carried out to produce sulphides or mercaptans.","Moreover, total immersion of the radiating portion of the lamp also makes it possible to remove, via the reaction medium, the heat emitted by the lamp; this arrangement simplifies the process and minimizes the equipment required (jacket, cooling-water circuit, exchangers, etc.).","However, this arrangement is incompatible with the use of a semi-continuous process in which the gradual introduction of at least one of the reagents entails an increase in the volume of the reaction medium in a reactor of fixed geometry.","SUMMARY OF THE INVENTION According to the invention, this problem has been solved by carrying out the photochemical process in a reactor comprising two zones, the radiating portion of the lamp being totally immersed in a first zone which is completely filled with reaction medium and spills off via an overflow into a second zone whose volume is sufficient to contain the volume of reaction medium originating from the first zone and corresponding substantially to the volume of the reagent(s) gradually introduced.","BRIEF DESCRIPTION OF THE DRAWINGS FIG.","1 shows the essential components of a reactor in accordance with the present invention.","FIG.","2 represents a device which is suitable for the semi-continuous manufacture of sulphides from maercaptan and an alkene in the presence of photoinitiator.","The invention will be understood more clearly in the light of the scheme in the attached FIG.","1 which shows essential components of a reactor in accordance with the present invention.","This reactor comprises a reaction zone (1) completely filled with reaction medium in which is at least totally immersed the radiating portion (2a) of a UV lamp (2) and, adjacent to the zone (1), a jacket zone (3) into which spills out the overflow from zone (1), pipes (4) and (5) for recycling, via at least one pump (6) and a heat exchanger (7), into the zone (1) via the distributor (9) the reaction medium after the additional reagent(s) has(have) been introduced therein via at least one pipe (8), a gassing vent (10) and a pipe (11) allowing the zone (1) of the reactor to be emptied.","In the preferred embodiment as represented in FIG.","1, the cylindrical zone (1) and the jacket zone (3) are coaxial.","However, since the inventive concept is to work in a reactor with two zones, one filled with reaction medium in which is totally immersed the radiating portion of the lamp, and the other collecting the overflow from the first zone, it would not constitute a departure from the context of the present invention to work in a reactor in which the two zones (1) and (3) were not coaxial.","In accordance with the present invention, the lamp (2) is installed in the zone (1) such that at least all of the radiating portion (approximately 65% of the length of the lamp) is immersed in the reaction medium, the top portion (about 35%) corresponding to the electrical portion (supply cable) not needing to be immersed.","However, it would not constitute a departure from the context of the present invention to immerse a greater portion of the lamp than that corresponding to the radiating portion.","As mentioned above, the volume of the zone (3) should be sufficient to contain the volume of reaction medium originating from the zone (1) and corresponding substantially to the volume of the reagent(s) gradually introduced.","The ratio between the volumes of the zones (1) and (3) thus depends on the reaction under consideration and may be calculated easily by a person skilled in the art from the reaction parameters (volume of zone 1, reaction time, volumes of the reagents, etc.).","The process and the device according to the invention have been designed for the photochemical manufacture of methyl ethyl sulphide (CH3SCH2CH3) from ethylene and methyl mercaptan, but they are also useful not only for the photochemical manufacture of other sulphides or mercaptans, but also more generally for any photochemical synthesis requiring the continuous supply of one or more reagents into the reactor during an operation.","The photochemical synthesis of sulphides and mercaptans by reacting an alkene with a mercaptan or hydrogen sulphide, optionally in the presence of a photoinitiator is generally carried out with a light source consisting of at least one low-pressure mercury lamp or any other source which delivers UV radiation of wavelengths between 200 and 400 nm.","This reaction is preferably performed at a pressure above atmospheric pressure which may range from 0.1 to 50 bar relative depending on the starting alkene and the starting mercaptan (or H2S); for the synthesis of methyl ethyl sulphide, the process is preferably performed at a pressure of between 2 and 10 bar relative.","As regards the reaction temperature, it is generally between −20 and 120° C. (preferably between 10 and 90° C.) and depends not only on the cooling system but also on the chosen working pressure.","For the manufacture of methyl ethyl sulphide, the ratio of the volume V1 of the zone (1) to the volume V3 of the zone (3) is generally between 0.4 and 0.7.The scheme in the attached FIG.","2 represents a device which is suitable for the semi-continuous manufacture of sulphides from a mercaptan and an alkene in the presence of a photoinitiator.","In this scheme, the inlets (8a), (8b) and (8c) are, respectively, those of the mercaptan, the photoinitiator and the alkene.","The operation consists firstly in filling the zone (1) with the mercaptan until it overflows into the annular zone (3) to a level (3a) which is sufficient to be able to start the pump (6) used to circulate and stir the reaction medium.","After loading the mercaptan via the pipe (8a), the pump (6) is switched on and the liquid originating from the zone (3) is cooled in the exchanger (7) and then conveyed to the zone (1).","Once the circulation is stable, the lamp (2) is switched on and, after a few minutes of establishing the working regime, the reaction starts with the simultaneous introduction of the photoinitiator via the pipe (8b) and of the alkene via the pipe (8c).","Since the volume of the reaction medium increases gradually with the introduction of the photoinitiator and of the alkene, the level in the zone (3) rises up to, for example, the level (3b), but the radiating portion of the lamp always remains totally immersed in the reaction medium and at the same time cooled thereby.","When the desired amount of mercaptan has been reacted, the supply of alkene and photoinitiator are stopped and the lamp is then switched off.","After degassing via the vent (10), the reaction medium is collected via the pipe (12) and optionally conveyed to a purification zone which, since it does not form the subject of the invention, is not represented herein.","The example which follows illustrates the invention without limiting it.","EXAMPLE In the device described in FIG.","2, methyl ethyl sulphide was prepared using, as light source, a low-pressure mercury lamp which re-emits by fluorescence radiation in the region of 350 nm.","This 58 watt lamp was placed axially in the zone (1) of the reactor having a total capacity of 50 litres, the volumes V1 and V3 being 15 and 35 litres, respectively.","23.2 kg of liquid methyl mercaptan were initially loaded so as to fill the zone (1) and to overflow into the zone (3).","Ethylene was introduced at a flow rate of 1.03 kg/hour and the photoinitiator (2,2-dimethoxy-2-phenylacetophenone as a solution at 200 g/l in methyl ethyl sulphide) was introduced at a flow rate such that the concentration of 2,2-dimethoxy-2-phenylacetophenone in the reaction medium was 0.1 g/litre.","The pressure in the reactor rose from 3 to 9 bar absolute; the temperature was maintained at 45° C. The supply of ethylene and photoinitiator was stopped after 13.1 hours and the reaction was left to continue while keeping the reaction medium in circulation for about one hour.","The amount of crude methyl ethyl sulphide collected after depressurization in the flow 12 was 35.95 kg.","At atmospheric pressure and at room temperature, this product had the following weight composition: Constituent % by weight Fractions 0.29 Methyl mercaptan 0.001 Ethyl mercaptan 0.004 Dimethyl sulphide 0.07 Methyl ethyl sulphide 99.57 Diethyl sulphide 0.002 Heavy fractions 0.06 The gaseous effluent, exiting via 10, weighed 0.76 kg and had the following weight composition: Constituent % by weight Fractions 39.3 Methyl mercaptan 60.6 From these results, the yield of methyl ethyl sulphide is established at 97% relative to the ethylene and at 98% relative to the methyl mercaptan."]],"string":"[\n [\n \"Semi-continuous photochemical method and device therefor\",\n \"In the photochemical synthetic process in semi-continuous mode according to the invention, a reactor comprising two zones is used, the radiating portion of the lamp(s) being totally immersed in a first zone which is completely filled with reaction medium and spills off via an overflow into a second zone whose volume is sufficient to contain the volume of reaction medium originating from the first zone and corresponding substantially to the volume of the reagent(s) gradually introduced.\"\n ],\n [\n \"1.Photochemical synthetic process in semi-continuous mode in which at least one of the reagents is introduced, from the start of the reaction, in total into the photochemical reactor, the other reagent(s) then being introduced gradually, characterized in that the process is performed in a reactor comprising two zones, the radiating portion of the lamp(s) being totally immersed in a first zone which is completely filled with reaction medium and spills off via an overflow into a second zone whose volume is sufficient to contain the volume of reaction medium originating from the first zone and corresponding substantially to the volume of the reagent(s) gradually introduced.\",\n \"2.Process according to claim 1, in which the reagent initially introduced in total into the first zone is a mercaptan or hydrogen sulphide, the reagent gradually introduced being an alkene.\",\n \"3.Process according to claim 1, in which a photoinitiator is also introduced gradually.\",\n \"4.Process according to claim 1, in which one or more lamp(s) delivering UV radiation of wavelengths between 200 and 400 nm is(are) used.\",\n \"5.Process according to one of claim 1, which is performed at a pressure ranging from 0.1 to 50 bar relative.\",\n \"6.Process according to one of claim 1, which is performed at a temperature of between −20 and 120° C. 7.Process according to claim 1, which is performed at a temperature of between 10 and 90° C. 8.Process according to claim 2, in which the mercaptan is methyl mercaptan and the alkene is ethylene.\",\n \"9.Process according to claim 1, in which the ratio of the volume of the first zone to the volume of the second zone is between 0.4 and 0.7.10.Process according to claim 5, which is performed at a pressure of between 2 and 10 bar relative.\",\n \"11.Device for carrying out the process according to claim 1, characterized in that it essentially comprises: a first zone 1 in which at least one lamp 2 is placed such that at least all of its radiating portion 2a is totally immersed, a second zone 3 adjacent to the zone 1 and serving as an overspill therefor, pipes 4 and 5 and a distributor 9 for ensuring, via at least one pump 6 and a heat exchanger 7, circulation of the reaction medium from the zone 3 to the zone 1, at least one pipe 8 for introducing the reagents, a degassing vent 10, and a discharge pipe 11.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>The expression “process performed in semi-continuous mode” means here a process in which at least one of the reagents is, from the start of the reaction, introduced in total into the photochemical reactor while at least one of the reagents is introduced gradually into the reactor as it is consumed.\",\n \"Such a procedure is occasionally found to be necessary in order to observe the optimum synthetic conditions.\",\n \"In a photochemical process operating in liquid medium, it is necessary for the radiating portion of the lamp to be constantly fully immersed in the liquid reaction medium.\",\n \"This total immersion is essential so as not to illuminate a gas phase whose behaviour, under ultraviolet irradiation, may prove to be hazardous or an inconvenience for the reaction under consideration.\",\n \"Thus, for example, in the synthesis of sulphides and mercaptans by photochemical reaction of an alkene with a mercaptan or hydrogen sulphide, UV irradiation of the gas phase may produce elemental sulphur which is a well-known inhibitor of free-radical reactions such as those carried out to produce sulphides or mercaptans.\",\n \"Moreover, total immersion of the radiating portion of the lamp also makes it possible to remove, via the reaction medium, the heat emitted by the lamp; this arrangement simplifies the process and minimizes the equipment required (jacket, cooling-water circuit, exchangers, etc.).\",\n \"However, this arrangement is incompatible with the use of a semi-continuous process in which the gradual introduction of at least one of the reagents entails an increase in the volume of the reaction medium in a reactor of fixed geometry.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>According to the invention, this problem has been solved by carrying out the photochemical process in a reactor comprising two zones, the radiating portion of the lamp being totally immersed in a first zone which is completely filled with reaction medium and spills off via an overflow into a second zone whose volume is sufficient to contain the volume of reaction medium originating from the first zone and corresponding substantially to the volume of the reagent(s) gradually introduced.\"\n ],\n [\n \"FIELD OF THE INVENTION The present invention relates to the field of photochemical processes and its subject is, more particularly, a photochemical process performed semi-continuously, and also a device for carrying it out.\",\n \"BACKGROUND OF THE INVENTION The expression “process performed in semi-continuous mode” means here a process in which at least one of the reagents is, from the start of the reaction, introduced in total into the photochemical reactor while at least one of the reagents is introduced gradually into the reactor as it is consumed.\",\n \"Such a procedure is occasionally found to be necessary in order to observe the optimum synthetic conditions.\",\n \"In a photochemical process operating in liquid medium, it is necessary for the radiating portion of the lamp to be constantly fully immersed in the liquid reaction medium.\",\n \"This total immersion is essential so as not to illuminate a gas phase whose behaviour, under ultraviolet irradiation, may prove to be hazardous or an inconvenience for the reaction under consideration.\",\n \"Thus, for example, in the synthesis of sulphides and mercaptans by photochemical reaction of an alkene with a mercaptan or hydrogen sulphide, UV irradiation of the gas phase may produce elemental sulphur which is a well-known inhibitor of free-radical reactions such as those carried out to produce sulphides or mercaptans.\",\n \"Moreover, total immersion of the radiating portion of the lamp also makes it possible to remove, via the reaction medium, the heat emitted by the lamp; this arrangement simplifies the process and minimizes the equipment required (jacket, cooling-water circuit, exchangers, etc.).\",\n \"However, this arrangement is incompatible with the use of a semi-continuous process in which the gradual introduction of at least one of the reagents entails an increase in the volume of the reaction medium in a reactor of fixed geometry.\",\n \"SUMMARY OF THE INVENTION According to the invention, this problem has been solved by carrying out the photochemical process in a reactor comprising two zones, the radiating portion of the lamp being totally immersed in a first zone which is completely filled with reaction medium and spills off via an overflow into a second zone whose volume is sufficient to contain the volume of reaction medium originating from the first zone and corresponding substantially to the volume of the reagent(s) gradually introduced.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS FIG.\",\n \"1 shows the essential components of a reactor in accordance with the present invention.\",\n \"FIG.\",\n \"2 represents a device which is suitable for the semi-continuous manufacture of sulphides from maercaptan and an alkene in the presence of photoinitiator.\",\n \"The invention will be understood more clearly in the light of the scheme in the attached FIG.\",\n \"1 which shows essential components of a reactor in accordance with the present invention.\",\n \"This reactor comprises a reaction zone (1) completely filled with reaction medium in which is at least totally immersed the radiating portion (2a) of a UV lamp (2) and, adjacent to the zone (1), a jacket zone (3) into which spills out the overflow from zone (1), pipes (4) and (5) for recycling, via at least one pump (6) and a heat exchanger (7), into the zone (1) via the distributor (9) the reaction medium after the additional reagent(s) has(have) been introduced therein via at least one pipe (8), a gassing vent (10) and a pipe (11) allowing the zone (1) of the reactor to be emptied.\",\n \"In the preferred embodiment as represented in FIG.\",\n \"1, the cylindrical zone (1) and the jacket zone (3) are coaxial.\",\n \"However, since the inventive concept is to work in a reactor with two zones, one filled with reaction medium in which is totally immersed the radiating portion of the lamp, and the other collecting the overflow from the first zone, it would not constitute a departure from the context of the present invention to work in a reactor in which the two zones (1) and (3) were not coaxial.\",\n \"In accordance with the present invention, the lamp (2) is installed in the zone (1) such that at least all of the radiating portion (approximately 65% of the length of the lamp) is immersed in the reaction medium, the top portion (about 35%) corresponding to the electrical portion (supply cable) not needing to be immersed.\",\n \"However, it would not constitute a departure from the context of the present invention to immerse a greater portion of the lamp than that corresponding to the radiating portion.\",\n \"As mentioned above, the volume of the zone (3) should be sufficient to contain the volume of reaction medium originating from the zone (1) and corresponding substantially to the volume of the reagent(s) gradually introduced.\",\n \"The ratio between the volumes of the zones (1) and (3) thus depends on the reaction under consideration and may be calculated easily by a person skilled in the art from the reaction parameters (volume of zone 1, reaction time, volumes of the reagents, etc.).\",\n \"The process and the device according to the invention have been designed for the photochemical manufacture of methyl ethyl sulphide (CH3SCH2CH3) from ethylene and methyl mercaptan, but they are also useful not only for the photochemical manufacture of other sulphides or mercaptans, but also more generally for any photochemical synthesis requiring the continuous supply of one or more reagents into the reactor during an operation.\",\n \"The photochemical synthesis of sulphides and mercaptans by reacting an alkene with a mercaptan or hydrogen sulphide, optionally in the presence of a photoinitiator is generally carried out with a light source consisting of at least one low-pressure mercury lamp or any other source which delivers UV radiation of wavelengths between 200 and 400 nm.\",\n \"This reaction is preferably performed at a pressure above atmospheric pressure which may range from 0.1 to 50 bar relative depending on the starting alkene and the starting mercaptan (or H2S); for the synthesis of methyl ethyl sulphide, the process is preferably performed at a pressure of between 2 and 10 bar relative.\",\n \"As regards the reaction temperature, it is generally between −20 and 120° C. (preferably between 10 and 90° C.) and depends not only on the cooling system but also on the chosen working pressure.\",\n \"For the manufacture of methyl ethyl sulphide, the ratio of the volume V1 of the zone (1) to the volume V3 of the zone (3) is generally between 0.4 and 0.7.The scheme in the attached FIG.\",\n \"2 represents a device which is suitable for the semi-continuous manufacture of sulphides from a mercaptan and an alkene in the presence of a photoinitiator.\",\n \"In this scheme, the inlets (8a), (8b) and (8c) are, respectively, those of the mercaptan, the photoinitiator and the alkene.\",\n \"The operation consists firstly in filling the zone (1) with the mercaptan until it overflows into the annular zone (3) to a level (3a) which is sufficient to be able to start the pump (6) used to circulate and stir the reaction medium.\",\n \"After loading the mercaptan via the pipe (8a), the pump (6) is switched on and the liquid originating from the zone (3) is cooled in the exchanger (7) and then conveyed to the zone (1).\",\n \"Once the circulation is stable, the lamp (2) is switched on and, after a few minutes of establishing the working regime, the reaction starts with the simultaneous introduction of the photoinitiator via the pipe (8b) and of the alkene via the pipe (8c).\",\n \"Since the volume of the reaction medium increases gradually with the introduction of the photoinitiator and of the alkene, the level in the zone (3) rises up to, for example, the level (3b), but the radiating portion of the lamp always remains totally immersed in the reaction medium and at the same time cooled thereby.\",\n \"When the desired amount of mercaptan has been reacted, the supply of alkene and photoinitiator are stopped and the lamp is then switched off.\",\n \"After degassing via the vent (10), the reaction medium is collected via the pipe (12) and optionally conveyed to a purification zone which, since it does not form the subject of the invention, is not represented herein.\",\n \"The example which follows illustrates the invention without limiting it.\",\n \"EXAMPLE In the device described in FIG.\",\n \"2, methyl ethyl sulphide was prepared using, as light source, a low-pressure mercury lamp which re-emits by fluorescence radiation in the region of 350 nm.\",\n \"This 58 watt lamp was placed axially in the zone (1) of the reactor having a total capacity of 50 litres, the volumes V1 and V3 being 15 and 35 litres, respectively.\",\n \"23.2 kg of liquid methyl mercaptan were initially loaded so as to fill the zone (1) and to overflow into the zone (3).\",\n \"Ethylene was introduced at a flow rate of 1.03 kg/hour and the photoinitiator (2,2-dimethoxy-2-phenylacetophenone as a solution at 200 g/l in methyl ethyl sulphide) was introduced at a flow rate such that the concentration of 2,2-dimethoxy-2-phenylacetophenone in the reaction medium was 0.1 g/litre.\",\n \"The pressure in the reactor rose from 3 to 9 bar absolute; the temperature was maintained at 45° C. The supply of ethylene and photoinitiator was stopped after 13.1 hours and the reaction was left to continue while keeping the reaction medium in circulation for about one hour.\",\n \"The amount of crude methyl ethyl sulphide collected after depressurization in the flow 12 was 35.95 kg.\",\n \"At atmospheric pressure and at room temperature, this product had the following weight composition: Constituent % by weight Fractions 0.29 Methyl mercaptan 0.001 Ethyl mercaptan 0.004 Dimethyl sulphide 0.07 Methyl ethyl sulphide 99.57 Diethyl sulphide 0.002 Heavy fractions 0.06 The gaseous effluent, exiting via 10, weighed 0.76 kg and had the following weight composition: Constituent % by weight Fractions 39.3 Methyl mercaptan 60.6 From these results, the yield of methyl ethyl sulphide is established at 97% relative to the ethylene and at 98% relative to the methyl mercaptan.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10473993"}}},{"rowIdx":779,"cells":{"text":{"kind":"list like","value":[["Modifying bitstreams","The bitstream includes digital codes representing an information signal.","At least one digital code is selected.","The code occupies a part of the bitstream which is to contain at least one watermark code which represents a watermark perceptible in the information signal.","The selected digital code(s) are removed from the said part of the bitstream.","The watermark code(s) are put in the said part of the bitstream in place of the selected code(s).","The number of bits of the selected code(s) removed from the said part of the bitstream is greater than or equal to the number of bits of the said watermark code(s) put in the said part.","The removed selected code(s) are appended to an end of the bitstream and/or placed in watermark user data fields created in the bitstream."],["1.A method of modifying a bitstream, the bitstream including digital codes representing information, the method comprising selecting at least one digital code which occupies a part of the bitstream which is to contain at least one modification code which represents a modification of the information, removing the selected digital code(s) from the said part of the bitstream, and putting the modified code(s) in the said part of the bitstream in place of the selected code(s), wherein the number of bits of the selected code(s) removed from the bitstream is greater than or equal to the number of bits of the said modification code(s) put into the bitstream; the method further comprising one or both of (1) storing the removed selected code(s) in association with the bitstream and (2) placing the removed selected codes in one or more removed code data fields in the bitstream.","2.A method according to claim 1, comprising redistributing the selected removed codes to a removed code data field and storing the removed selected codes in association with the bitstream in a preferred order, which is: 1) within the removed code data field; and 2) storing the removed selected codes in association with the bitstream.","3.A method according to claim 1, comprising the steps of both storing some of the removed selected codes in association with the bitstream and placing others of the removed selected codes in the said one or more removed code data fields.","4.A method according to claim 1, wherein the step of (1) storing the removed selected code(s) in association with the bitstream comprises appending the removed selected code(s) to an end of the bitstream.","5.A method according to claim 1, wherein the step of (1) storing the removed selected code(s) in association with the bitstream comprises storing the removed selected codes in a file.","6.A method according to claim 5, wherein the file is appended to an end of the bitstream.","7.A method according to claim 6, wherein the file is appended to the start end of the bitstream.","8.A method according to claim 5, wherein the file is separate from, and associated with, the bitstream.","9.A method according to claim 1, comprising the step of appending all the removed selected code(s) to an end of the bitstream.","10.A method according to claim 1, comprising the step of placing all the removed selected code(s) in the said one or more removed code data fields.","11.A method according to claim 10, comprising the step of compressing an information signal to produce the said bitstream, the amount of compression being based on a prediction of the amount of data in the modified bitstream to allow all the removed selected codes to be placed in removed code data fields within the bitstream.","12.A method according to claim 1, comprising the step of adding padding bits to the modified bitstream so that the bitrate of the modified bitstream is unchanged relative to the original bitstream.","13.A method according to claim 1, comprising the step of using a template to define the part or parts of the bitstream which is/are to contain modification code(s).","14.A method according to claim 1 wherein the said digital codes representing the information are codes representing transform coefficients of the information.","15.A method according to claim 14, wherein the coefficients are DCT coefficients.","16.A method according to claim 15, wherein the DCT coefficients include DC and AC coefficients, all the codes representing the DC and AC coefficients are removed from the said part of the bitstream and the modification codes placed in the said part of the bitstream are codes representing DC coefficients.","17.A method according to claim 16, wherein the codes removed from the same part of the bitstream are grouped in a group identified by a group code.","18.A method according to claim 17, wherein the said part of the bitstream is at least one DCT block.","19.A method according to claim 16, wherein the bitstream is modifiable by: at least one of 1) the technique of removing codes representing AC coefficients and 2) the technique of removing codes representing DC coefficients: replacing the DC codes by modified DC codes and storing the removed selected code(s) (a) in association with the bitstream and/or (b) placing them in one or more removed code data fields in the bitstream: the method further comprising the step of assessing the effects of the said techniques on the number of bits in the bitstream and selecting, in dependence on the assessment, one or more of the said techniques for performing the modification of the bitstream.","20.A method according to claim 15, wherein the bitstream is a video bitstream which comprises at least intra encoded pictures and the method is performed on the intra-encoded pictures.","21.A method according to claim 20, wherein the bitstream comprises inter-encoded pictures and the method is performed on the inter-encoded pictures.","22.A method according to claim 21, wherein the inter-encoded pictures include DC codes representing DC coefficients, and comprising the steps of removing selected ones of the said DC codes, storing them in association with the bitstream and/or placing them in one or more removed code data fields in the bitstream and replacing the removed codes with modification codes of predetermined value.","23.A method according to claim 22, wherein the said DC codes representing DC coefficients in inter-encoded pictures represent quantised change values and the said predetermined value represents the minimum quantum of change greater than zero.","24.A method according to claim 23, wherein the said predetermined value is plus or minus one.","25.A method according to claim 21, wherein the inter-encoded pictures include codes representing motion vectors and further comprising the steps of removing the motion vector codes associated with parts of the bitstream which are to contain modification codes, replacing the original motion vector codes with motion vector modification codes representing zero motion vectors, and appending the removed motion vector codes to the end of the bitstream and/or placing the removed codes in removed code data fields in the bitstream.","26.A method according to claim 25, comprising the further steps of removing the motion vector codes associated with parts of the inter-encoded pictures which, in the picture, are adjacent the said parts of the bitstream which are to contain modification codes, replacing the original motion vector codes of the said associated parts with motion vector modification codes representing zero motion vectors, and appending the removed motion vector codes to the end of the bitstream and/or placing the removed codes in removed code data fields in the bitstream.","27.A method according to claim 25, comprising providing a first template defining the blocks of intra-encoded pictures to be modified and a second template defining the blocks corresponding to the blocks of the first template of the inter-encoded pictures for which the associated motion vectors are to be set to zero.","28.A method according to claim 27, wherein the second template defines macroblocks which include the said blocks for which the associated motion vectors are to be set to zero.","29.A method according to claim 28, wherein the second template defines further macroblocks, for which the associated motion vectors are to be set to zero, bordering the said first-mentioned macroblocks.","30.A method according to claim 20, wherein the bitstream comprises DCT blocks which may be encoded as field type or frame type with type codes indicating the type of encoding, and comprising the step of removing selected ones of the said type codes indicating field type and replacing the removed type codes with modification type codes indicating frame type and storing the removed type codes in association with the bitstream and/or placing them in one or more removed code data fields in the bitstream.","31.A method according to claim 15, wherein the bitstream is an MPEG2 bitstream.","32.A method according to claim 1, comprising the further step of providing data indicating portions of the bit stream which contain modification codes.","33.A method according to claim 31, wherein the bitstream includes a supplementary data field, and comprising the step of providing data in the bitstream which distinguishes the removed code data field(s) from the said supplementary data field.","34.A method according to claim 1, comprising the step of encrypting the removed selected codes before storing them in association with the bitstream and/or placing them in the said removed code data field(s).","35.A method of restoring an original bitstream, the method comprising receiving a modified bitstream which has modification codes in selected parts of the bitstream from which original codes have been removed and stored, the method comprising the further steps of: parsing the bitstream; taking the original codes from store; and replacing the modification codes with the original codes.","36.A method of restoring an original bitstream, the method comprising receiving a modified bitstream which has modification codes in selected parts of the bitstream from which original codes have been removed and placed in modification user data portions of the bitstream, the method comprising the further steps of: parsing the bitstream; storing the original codes; and replacing the modification codes with the stored original codes.","37.A method of restoring an original bitstream, the method comprising receiving a modified bitstream which has modification codes in selected parts of the bitstream from which original codes have been removed and placed in modification user data portions of the bitstream and appended to an end of the bitstream, the method comprising the further steps of: parsing the bitstream; storing the original codes; and replacing the modification codes with the stored original codes.","38.A method according to claim 36, wherein the original codes are encrypted and comprising the further step of decrypting them before replacing the modification codes with the decrypted original codes.","39.A method according to claim 35, comprising the step of using a template which defines the part or parts of the bitstream which contain the modification code(s) to select those codes for replacement.","40.A method according to claim 1, wherein the modification creates a perceptible effect in the information represented by the information signal.","41.Apparatus arranged to carry out the method of claim 1.42.A computer program arranged to carry out the method of claim 1, when run on a suitable processor.","43.A transmission medium storing a computer program according to claim 42.44.A medium according to claim 43 which is a storage medium.","45.A medium according to claim 43 which is a communications channel.","46.A modified bitstream which has modification codes in selected parts of the bitstream from which original codes have been removed and appended to an end of the bitstream.","47.A modified bitstream which has modification codes in selected parts of the bitstream from which original codes have been removed and placed in modification user data portions within the bitstream.","48.A modified bitstream which has modification codes in selected parts of the bitstream from which original codes have been removed and some placed in modification user data portions within the bitstream and others appended to an end of the bitstream.","49.Apparatus for modifying a bitstream, the bitstream including digital codes representing information, the apparatus comprising a selecting stage operable to select at least one digital code which occupies a part of the bitstream which is to contain at least one modification code which represents a modification perceptible in the information, a removal stage operable to remove the selected digital code(s) from the said part of the bitstream, a modification code placing stage operable to put the modification code(s) in the said part of the bitstream in place of the selected code(s), wherein the number of bits of the selected code(s) removed from the bitstream is greater than or equal to the number of bits of the said modification code(s) put into the bitstream, and a removed code placing stage operable to (1) store the removed selected code(s) in association with the bitstream and/or (2) place the removed selected codes in one or more removed code data fields in the bitstream.","50.Apparatus according to claim 49, wherein the removed code placing stage is operable to redistribute the selected removed codes to a removed code data field and store the removed selected codes in association with the bitstream in a preferred order, which is: 1) within the removed code data field; and 2) store the removed selected codes in association with the bitstream.","51.Apparatus according to claim 49, wherein the removed code placing stage is operable to both store some of the removed selected codes in association with the bitstream and place others of the removed selected codes in the said one or more removed code data fields.","52.Apparatus according to claim 49, wherein the removed code placing stage is operable to (1) store the removed selected code(s) in association with the bitstream by appending the removed selected code(s) to an end of the bitstream.","53.Apparatus according to claim 49, wherein the removed code placing stage is operable to (1) store the removed selected code(s) in association with the bitstream by storing the removed selected codes in a file.","54.Apparatus according to claim 53, wherein the removed code placing stage is operable to append the file to an end of the bitstream.","55.Apparatus according to claim 53, wherein the removed code placing is operable to store the file separate from, but associated with, the bitstream.","56.Apparatus according to claim 49, wherein the removed code placing stage is operable to append all the removed selected code(s) to an end of the bitstream.","57.Apparatus according to claim 49, wherein the removed code placing stage is operable to place all the removed selected code(s) in the said one or more removed code data fields.","58.Apparatus according to claim 49, wherein the said digital codes represent DCT coefficients of the said information.","59.Apparatus according to claim 58, wherein the DCT coefficients include DC and AC coefficients, the removal stage is operable to remove all the codes representing the DC and AC coefficients from the said part of the bitstream and the modification code placing stage is operable to place the modification codes representing DC coefficients in the said part of the bitstream.","60.Apparatus according to claim 58, wherein the removal stage is operable to carry out at least one of 1) a technique to remove codes representing AC coefficients and 2) a technique to remove codes representing DC coefficients: the modification code placing stage is operable to replace the removed DC codes by modified DC codes, and the removed code placing stage is operable to store the removed selected code(s) in association with the bitstream and/or place them in one or more removed code data fields in the bitstream, the apparatus further comprising a stage operable to assess the effects of the said techniques on the number of bits in the bitstream and select one or more of the said techniques in dependence on the assessment.","61.Apparatus according to claim 60 wherein the removed code placing stage is operable to append the removed selected code(s) to an end of the bitstream and/or place them in one or more removed code data fields in the bitstream.","62.Apparatus according to claim 59, wherein the bitstream comprises at least intra encoded pictures and the apparatus is arranged to modify the intra-encoded pictures.","63.Apparatus according to claim 62, wherein the bitstream comprises inter-encoded pictures and the apparatus is arranged to additionally modify the inter-encoded pictures.","64.Apparatus according to claim 63, wherein the inter-encoded pictures include codes representing DC coefficients, the apparatus comprising a stage operable to remove selected ones of the said codes, and a placing stage operable to store them in association with the bitstream and/or place them in one or more removed code data fields in the bitstream and a replacing stage operable to replace the removed codes with modification codes of predetermined value.","65.Apparatus according to claim 64, wherein the said codes representing DC coefficients in inter-encoded pictures represent quantised change values and the said predetermined value represents the minimum quantum of change greater than zero.","66.Apparatus according to claim 65, wherein the said predetermined value is plus or minus one.","67.Apparatus according to claim 63, wherein the inter-encoded pictures include codes representing motion vectors and further comprising a motion vector code removing stage operable to remove the motion vector codes associated with parts of the bitstream which are to contain modification codes, replace the original motion vector codes with motion vector modification codes representing zero motion vectors, and store the removed motion vector codes in association with the bitstream and/or placing the removed codes in removed code data fields in the bitstream.","68.Apparatus according to claim 67, wherein the motion vector code removing stage is further operable to remove the motion vector codes associated with parts of the inter-encoded pictures which, in the picture, are adjacent the said parts of the bitstream which are to contain modification codes, replace the original motion vector codes of the said associated parts with motion vector modification codes representing zero motion vectors, and store the removed motion vector codes in association with the bitstream and/or place the removed codes in removed code data fields in the bitstream.","69.Apparatus according to claim 58, wherein the bitstream comprises DCT blocks which may be encoded as field type or frame type with type codes indicating the type of encoding, and comprising a removal stage operable to remove selected ones of the said type codes indicating field type and a replacing stage operable to replace the removed type codes with modification type codes indicating frame type and a removed code placing stage operable to store the removed type codes in association with the bitstream and/or place them in one or more removed code data fields in the bitstream.","70.Apparatus for restoring an original bitstream, the apparatus comprising a receiving stage operable to receive a modified bitstream which has modification codes in selected parts of the bitstream from which original codes have been removed and stored, a parsing stage operable to parse the bitstream; a store operable to store the original codes; and a replacing stage operable to replace the modification codes with the original codes.","71.Apparatus for restoring an original bitstream, the apparatus comprising a receiving stage operable to a receive a modified bitstream which has modification codes in selected parts of the bitstream from which original codes have been removed and placed in modification user data portions of the bitstream: a parsing stage operable to parse the bitstream; a store for storing the original codes; and a replacing stage operable to replace the modification codes with the stored original codes.","72.Apparatus for restoring an original bitstream, comprising a receiving stage operable to receive a modified bitstream which has modification codes in selected parts of the bitstream from which original codes have been removed and placed in modification user data portions of the bitstream and appended to an end of the bitstream, a parsing stage operable to parse the bitstream; a store for storing the original codes; and a replacing stage operable to replace the modification codes with the stored original codes.","73.Apparatus according to claim 71, for use with a bitstream wherein the original codes are encrypted and comprising a decrypting stage operable to decrypt the original codes before replacing the modification codes with the decrypted original codes.","74.A data carrier storing a template defining the part or parts of a bitstream which is/are to contain modification code(s) and at least one decryption key.","75.The invention of claim 74 wherein the information signal includes one or more of: video data; audio data; audio/visual data; and other data.","76.A method of visibly watermarking a video sequence having intra-encoded pictures and inter-encoded pictures, the pictures comprising data representing blocks of transform coefficients of the video, motion vectors being associated with at least some of the blocks of the inter-encoded pictures, the method comprising providing at least one template defining the blocks to be modified to visibly watermark the video, modifying at least some transform coefficients in the blocks defined by the template(s) in at least the intra-encoded pictures to visibly watermark the video, and setting to zero the motion vectors associated with blocks of the inter-encoded pictures defined by the template(s) and corresponding to the modified blocks of the intra-encoded pictures.","77.A method according to claim 76, comprising providing a first template defining the blocks of intra-encoded pictures to be modified and a second template defining the blocks corresponding to the blocks of the first template of the inter-encoded pictures for which the associated motion vectors are to be set to zero.","78.A method according to claim 77, wherein the second template defines macroblocks which include the said blocks for which the associated motion vectors are to be set to zero.","79.A method according to claim 78, wherein the second template defines further macroblocks, for which the associated motion vectors are to be set to zero, bordering the said first-mentioned macroblocks.","80.A method according to claim 76, wherein the video sequence is represented by a bitstream including digital codes representing the blocks and motion vectors, the method comprising selecting the digital codes which represent the blocks to be modified and the motion vectors to be set to zero, removing the selected digital code(s) from the bitstream, and putting modified codes in the bitstream in place of the selected codes, the method further comprising one or both of (1) appending the removed codes to an end of the bitstream and (2) placing the removed codes in one or more removed code data fields in the bitstream.","81.A method according to claim 80, further comprising encrypting the removed codes.","82.Apparatus arranged to carry out the method of claim 76.83.A computer program which when run on a suitable data processor implements the method of claim 76.84.A data carrier on which is stored a program according to claim 83.85.Apparatus for visibly watermarking a video sequence having intra-encoded pictures and inter-encoded pictures, the pictures comprising data representing blocks of transform coefficients of the video, motion vectors being associated with at least some of the blocks of the inter-encoded pictures, the apparatus comprising a storage means for storing at least one template defining the blocks to be modified to visibly watermark the video, a modifier arranged to modify at least some transform coefficients in the blocks defined by the template(s) in at least the intra-encoded pictures to visibly watermark the video, and a means arranged to set to zero the motion vectors associated with blocks of the inter-encoded pictures defined by the template(s) and corresponding to the modified blocks of the intra-encoded pictures.","86.Apparatus according to claim 85, wherein the storage means stores a first template defining the blocks of intra-encoded pictures to be modified and a second template defining the blocks corresponding to the blocks of the first template of the inter-encoded pictures for which the associated motion vectors are to be set to zero.","87.Apparatus according to claim 86, wherein the second template defines macroblocks which include the said blocks for which the associated motion vectors are to be set to zero.","88.Apparatus according to claim 87, wherein the second template defines further macroblocks, for which the associated motion vectors are to be set to zero, bordering the said first-mentioned macroblocks.","89.Apparatus according to claim 85, wherein the video sequence is represented by a bitstream including digital codes representing the blocks and motion vectors, the apparatus comprising means for selecting the digital codes which represent the blocks to be modified and the motion vectors to be set to zero, means for removing the selected digital code(s) from the bitstream, and putting modified codes in the bitstream in place of the selected codes, and means for (1) appending the removed codes to an end of the bitstream and/or (2) placing the removed codes in one or more removed code data fields in the bitstream.","90.A method of restoring an original bitstream visibly watermarked by the method of claim 80, the modified bitstream having modified codes representing modified blocks and motion vectors set to zero in selected parts of the bitstream from which original codes have been removed and placed in modification user data portions of the bitstream and/or appended to the end of the bitstream, the method comprising the further steps of: parsing the bitstream; storing the original codes; and replacing the modified codes with the stored original codes.","91.A method according to claim 90, wherein the original codes are encrypted and comprising the further step of decrypting them before replacing the modification codes with the decrypted original codes.","92.Apparatus for restoring an original bitstream, comprising: a receiving stage operable to receive a modified bitstream visibly watermarked by the method of claim 5, the modified bitstream having modified codes representing modified blocks and motion vectors set to zero in selected parts of the bitstream from which original codes have been removed and placed in modification user data portions of the bitstream and/or appended to the end of the bitstream the apparatus comprising a parsing stage operable to parse the bitstream; a store operable to store the original codes; and a replacing stage operable to replace the modified codes with the original stored codes.","93.Apparatus according to claim 92, wherein the original codes are encrypted and comprising the further a stage for decrypting them before replacing the modified codes with the decrypted original codes.","94.Apparatus arranged to carry out the method of claim 90.95.A computer program which when run on a suitable data processor implements the method of claim 90.96.A data carrier on which is stored a program according to claim 95.97.","(canceled) 98.","(canceled) 99.","(canceled) 100.","(canceled)"],["The present invention relates to a method of modifying a bitstream, apparatus for modifying a bitstream, a computer program for modifying a bitstream, a storage medium storing such a program, method of removing a modification from a bitstream, apparatus for removing a modification from a bitstream, a computer program for removing a modification from a bitstream, a storage medium storing such a program, a modified bitstream and a data carrier carrying modification removal data.","It is known to provide a watermark in video, audio, audio/visual and/or other information signals.","The watermark is provided to check the provenance of the information signal and/or to identify the owner or other persons associated with the information signal.","A watermark may be perceptible or hidden.","Preferred embodiments of the present invention use perceptible watermarks.","It is also known to watermark an image by transforming the image from the spatial domain to a transform domain, e.g.","to the wavelet domain and embed a watermark by changing the wavelet coefficients.","The transform domain image with the watermark is then inverse transformed to the spatial domain.","See for example “An image watermarking method based on the wavelet transform” by Hisashi Inoue, et al, IEEE, 0-7803-5467-2/99.U.S.","Pat.","No.","5,809,139 (Girod et al) discloses the invisible watermarking of video material which is already compressed.","Video which has been compressed by MPEG2 is entropy decoded and dequantised to obtain the DCT coefficients.","A spread spectrum spatial domain watermark is transformed to the DCT domain and the transformed watermark added to the DCT coefficients of the transformed video.","To maintain the bit rate of the compressed video even with the addition of the watermark, various steps are taken.","Only non-zero coefficients are modified.","Also the number of bits of an encoded watermarked coefficient is compared against the number of bits of the coefficient prior to watermarking (plus any extra number of spare bits saved in previous coding operations).","A control controls selection of the output between the watermarked bitstream and the original unwatermarked bitstream.","If an entropy encoded unwatermarked coefficient uses n0 bits and the watermarked entropy encoded coefficient uses n1 bits then a watermarked coefficient is output if n1<=n0+n3, where n3 is the number of spare bits available for use.","The watermark can be detected in a video decoder but it is not possible to restore the watermarked video to the original video by reversing the watermarking algorithm.","WO 99/10837 (Digimarc) discloses a method of watermarking a video image in which the image is compressed using for example MPEG2 and the DCT coefficients are modified to embed a watermark.","The bit rate is preserved by maintaining a count that represents the amount the bit rate has been increased by the modifications to the coefficients less the amount the bit rate has been decreased by the modifications.","If the cumulative change exceeds a positive or negative limit, then no further changes are made.","The watermark can be detected in a video decoder but it is not possible to restore the watermarked video to the original video by reversing the watermarking algorithm.","Both U.S. Pat.","No.","5,809,139 and WO 99/10837 require means which monitor the bit rate and which stops embedding of a watermark if a specified limit is reached.","For US-A-5809139 and WO 99/10837 that results in a random distribution of the watermark in an image.","These proposals are aimed at irreversible imperceptible watermarks which embed data representing a message in the image.","It is known to compression encode information signals especially image signals.","Also, if a modification is made to the information signals which adds data to the information signals, then the compression is usually less efficient.","It is desirable to process compressed information signals, so as to produce modification of the information signals which is perceptible in the decompressed information signals, e.g.","a visible modification in an image, with a minimal change to the number of bits representing the compressed information signals.","Copending prior application EP 01310113.4 (U.S. Ser.","No.","10/006,050) is an example of a method and apparatus for modifying a compressed video bitstream and for removing the watermark therefrom.","A compression process carried out on the bitstream uses DCT coefficients.","The modifying process is carried out to ensure the watermarked bitstream has no more bits than, and thus the same bit rate as, the bitstream before it was watermarked.","That is done by encrypting fixed length parts of codes representing DCT coefficients without increasing the number of bits in the fixed length codes.","The most preferred embodiment is a compressed bitstream of I frames and the technique works well with such a bitstream to produce a visible watermark.","The technique can be applied to other compressed bitstreams but the visibility of the watermark in the displayed image may not be as desired due to the limitation of not increasing the number of bits.","MPEG2 is well known and is defined in ISO/IEC/13818.The invention seeks to provide such a technique which is applicable to bitstreams containing video, audio, audio/visual and/or other information signals and which produces a modification, preferably a removable and perceptible modification, without increasing the bitrate, and preferably without changing the bit rate, of the information signal.","An embodiment of the present invention seeks to provide a modifying technique which can be applied to a compressed bitstream, (e.g.","MPEG2) and which can produce a modification of desired perceptibility, whilst at the same time not changing the bit rate in the compressed bitstream and which modification is removable from the bitstream.","One aspect of the invention provides a method of modifying a bitstream, the bitstream including digital codes representing information, the method comprising selecting at least one digital code which occupies a part of the bitstream which is to contain at least one modification code which represents a modification of the information, removing the selected digital code(s) from the said part of the bitstream, and putting the modified code(s) in the said part of the bitstream in place of the selected code(s), wherein the number of bits of the selected code(s) removed from the bitstream is greater than or equal to the number of bits of the said modification code(s) put into the bitstream; the method further comprising one or both of (1) storing the removed selected code(s) in association with the bitstream and (2) placing the removed selected codes in one or more removed code data fields in the bitstream.","Preferably, the modification is such as to be perceptible in the information represented by the bitstream.","According to a second aspect of the present invention there is provided a method of modifying a bitstream, the bitstream including digital codes representing information, the method comprising selecting at least one digital code which occupies a part of the bitstream which is to contain at least one modifying code which represents a modification perceptible in the information, removing the selected digital code(s) from the said part of the bitstream, putting the modification code(s) in the said part of the bitstream in place of the selected code(s), wherein the number of bits of the selected code(s) removed from the bitstream is greater than or equal to the number of bits of the said modification code(s) put in the said part, and appending the removed selected code(s) to an end of, but not within, the bitstream According to a third aspect of the present invention, there is provided a method of modifying a bitstream having an information signal portion including digital codes representing information, the method comprising selecting digital codes which occupy a part of the said information signal portion which is to contain at least one modification code which represents a modification perceptible in the information, removing the selected digital codes from the said part of the bitstream, putting the modification code(s) in the said part of the bitstream in place of the selected codes, creating a modification user data portion of the bitstream appended with the said part and storing at least one of the removed selected code(s) in the said modification user data portion, wherein the number of bits in the said information signal portion containing the modification codes plus the number of bits in the said modification user data portion is less than or equal to the number of bits in the said information signal portion before removal of the selected codes.","The modification in embodiments of the invention is the provision of a perceptible modification of the information represented by bitstream rather than the hiding of data within the bitstream.","In this way, the bitrate of the modified bitstream is maintained equal to that of the original bitstream.","The addition of the modification codes produces additional data.","By removing the original codes and appending them to the end of the bitstream, the amount of data in the bitstream is not increased.","Spare space created in the bitstream by the removal of the original codes may be used as the modification user data fields for at least some of the original codes and any codes which cannot be included in those fields are appended to an end of the bitstream.","The codes may be appended to the start end of the bitstream or to the finish end.","The codes may be in a separate file or in a file appended to the start end or the finish end of the bitstream.","The bitstream in preferred examples of the invention is a compressed bitstream preferably compressed according to the MPEG2 standard.","It is particularly important in MPEG2 that the bitrate of the bitstream conforms to the standard.","Thus the present invention makes possible that the MPEG2 bitstream even with the modification codes in it complies with the standard and any excess data resulting from the modifying process is not in the MPEG2 compliant bitstream but appended to it.","Excess data is the data which cannot be placed in the modification user data fields in the bitstream.","Preferred embodiments of the present invention allow all the original data to be preserved allowing removal of the modification data and restoration of the original bitstream.","(This assumes, of course, that data is not lost due to other processing of the modified bitstream for example transmission via a noisy channel or further processing of the modified signal.)","A fourth aspect of the invention provides an apparatus for modifying a bitstream, the bitstream including digital codes representing information, the apparatus comprising a selecting stage operable to select at least one digital code which occupies a part of the bitstream which is to contain at least one modification code which represents a modification perceptible in the information, a removal stage operable to remove the selected digital code(s) from the said part of the bitstream, a modification code placing stage operable to put the modification code(s) in the said part of the bitstream in place of the selected code(s), wherein the number of bits of the selected code(s) removed from the bitstream is greater than or equal to the number of bits of the said modification code(s) put into the bitstream, and a removed code placing stage operable to (1) store the removed selected code(s) in association with the bitstream and/or (2) place the removed selected codes in one or more removed code data fields in the bitstream.","In an embodiment of the invention the bitstream comprises digital codes representing transform coefficients of the information signal, for example DCT coefficients, including DC and AC coefficients.","An example of the invention uses various techniques for modifying such a bitstream.","Those techniques include 1) removing AC coefficients and 2) removing DC coefficients.","The removed coefficients are replaced by modification codes.","The effects of the various techniques on the number of bits in the modified bitstream is assessed and one or more of the techniques is selected for the actual modification of the bitstream in dependence on the assessment.","This allows the techniques to be selected to maintain the bitrate of the modified bit stream equal to that of the original bitstream.","The original bitstream may include inter-encoded pictures including codes representing motion vectors.","If the modification represents a visible watermark in a picture, the motion vectors distort the watermark.","An example of the invention, comprises the steps of removing the motion vector codes associated with the said part of the bitstream which is to contain at least one modification code, replacing the original motion vector codes with motion vector modification codes representing zero motion vectors, and placing the removed motion vector codes at the end of the bitstream and/or in removed code data fields in the bitstream.","This reduces distortion of the watermark.","MPEG2 bitstreams may include DCT blocks encoded as field type or frame type with type codes indicating the type of encoding.","It has been found that if a field type DCT block is modified but an adjacent field type block is not modified the watermark is distorted by stripes.","Thus an example of the invention comprises the step of removing selected ones of the said type codes indicating field type and replacing the removed type codes with modification type codes indicating frame type and placing the removed type codes in one or both of (1) the end of the bitstream and (2) in one or more removed code data fields in the bitstream.","Other aspects of the invention are specified in the claims to which attention is invited.","For a better understanding of the present invention, and to show how the same may be carried into effect, reference will now be made, by way of example, to the accompanying drawings in which: FIG.","1 is a schematic diagram of an image frame; FIG.","2 is a schematic diagram of blocks of 8×8 pixels in the image frame; FIG.","3 is a schematic diagram of a DCT block; FIG.","4 is a schematic diagram of macroblocks; FIG.","5 is a schematic diagram of rows of macroblocks and slices; FIG.","6 is a schematic diagram of a Group of Pictures (GOP); FIG.","7 is a schematic diagram of one frame of picture data; FIG.","8 is a schematic diagram of a bitstream of one macroblock; FIG.","9 is a schematic diagram of a template defining a watermark; FIG.","10 is a flow diagram illustrating an overview of an illustrative watermarking method applied to intra-frames according to the present invention; FIGS.","11 to 13 are schematic diagrams of illustrative data formats used in examples of the present invention; FIG.","14 is schematic diagram showing user data distributed amongst compressed picture frames; FIG.","15 is a flow diagram of a watermark encryption/decryption process; FIG.","16 is a flow diagram illustrating an overview of an illustrative watermark removal method according to the present invention; FIG.","17 is an overview of an illustrative watermarking system according to the present invention; FIG.","18 is a schematic block diagram of a watermark removal apparatus; FIG.","19 is a schematic diagram of three buffers used in the watermarking process and in the watermark removal process; FIG.","20 is a flow diagram illustrating an overview of an illustrative watermarking method applied to non-intra-frames according to the present invention; FIG.","21A illustrates watermarked macroblocks and FIG.","21B is a flow diagram of a DCT type change process; FIG.","22 is a flow diagram of a technique selection process; FIG.","23 illustrates templates used in association with Motion Vectors; FIG.","24 is a flow diagram illustrating an overview of an illustrative method applied to motion vector codes in an embodiment of the present invention; FIG.","25 illustrates the coding of motion vectors; FIG.","26 illustrates watermarking a video sequence of frame pictures; FIG.","27 illustrates a) watermarking a video sequence of field pictures, and b) watermarking a video sequence of field and frame pictures; and FIG.","28 illustrates watermarked macroblocks for interlaced fields and for a frame; FIGS.","29 to 31 illustrates modifications of the invention.","In the following illustrative description, an MPEG2 compressed bitstream is modified.","The modification is referred to as a watermark.","Applying the watermark to the bitstream is referred to as embedding.","Restoring the original bitstream is referred to as washing.","FIRST EXAMPLE A first example of the present invention will now be described.","For ease of explanation, the example describes the placing of watermark data into I frames of an MPEG2 video bitstream which is a single stream with no audio; i.e.","an MPEG2 video sequence file.","Alternatively, the bitstream may be a packetised bitstream.","However the invention is not limited to I frames nor is it limited to video nor is it limited to MPEG2.Also for ease of explanation, the following discussion refers only to luminance information.","However the invention is not limited to luminance information.","Firstly, MPEG2 will be reviewed.","Referring to FIG.","1, an image frame 2 is shown.","The image is represented by digital pixels.","As shown in FIG.","2 blocks 4 of 8×8 pixels are selected and as shown in FIG.","3 those blocks are subject to a Discrete Cosine Transform (DCT) to form DCT blocks containing 8×8 DCT coefficients.","The coefficients include a DC coefficient 6 and AC coefficients 7.To form a serial bitstream the coefficients are scanned in a zig-zag scan pattern 8 as illustrated in FIG.","3.","(Other patterns are known).","Groups of four adjacent DCT blocks are grouped as macroblocks 10 as shown in FIG.","4.As shown in FIG.","5, the image frame is organised as rows R1, R2, R3 etc.","of macroblocks.","The macroblocks are organised into slices.","A slice is a horizontal collection of macroblocks.","A new slice starts at the beginning of each row of macroblocks.","A slice may contain any number of macroblocks.","A row of macroblocks may contain any number of slices.","Referring to FIG.","6, MPEG2 has the following types of frame.","An I frame is an intra encoded frame; that is a frame in which all the image data is contained in that frame without reference to any other frame.","A P frame is a frame that is coded using motion compensated prediction from a previous I frame or P frame.","A B frame is a frame coded using motion compensated prediction from a previous and/or future I frame or P frame.","For ease of explanation the following description of the first example refers only to I frames.","Frames are organised into Groups of Pictures called GOPs.","FIG.","6 shows part of a typical GOP in which a repeated sequence of I, B, B and P frames form the GOP.","Other sequences are possible.","The number of frames in a typical GOP is 12 or 15, but other numbers of frames are possible.","FIG.","7 and FIG.","11 illustrates the data format of what is referred to herein as picture data.","Picture data herein includes all data between picture start codes and includes headers (HDR), userdata (UD) and the image data 16 representing one frame of the image.","MPEG2 allows user data (UD) to be placed in the picture data ahead of the image data 16.There may be no user data fields UD or one or more user data fields.","The DCT blocks and macroblocks are formatted as shown in FIG.","8 in the image data 16.Each block includes a DC coefficient followed by AC coefficients.","Blocks are separated by End of Block codes EOB.","In I frames the DC coefficients are differentially encoded.","The differential values are referred to herein as DC differentials.","The AC coefficients are run length encoded and represented by Variable Length Codes (VLCs) and are referred to herein as AC VLCs.","The MPEG2 encoding process must produce a bitstream which is decodable by all decoders which comply with the MPEG2 standard.","Decoders include a buffer which temporarily stores the bitstream as it is decoded in the buffer.","The MPEG2 standard defines the size of the buffer.","Generally, the buffer should neither underflow nor overflow otherwise data is lost.","(Underflow may be allowed under certain conditions) The encoding process is carried out such that the bit rate is constrained to prevent underflow and overflow.","Modifying an encoded bitstream, in this example of the invention for the purpose of applying a visible watermark to it, risks changing the bit rate and thus risks underflow or overflow.","The example of the invention described hereinafter seeks to apply a visible watermark to the bitstream without changing the bit rate of the picture data and thus without causing the buffer to underflow or overflow and to allow the watermark to be removed and the original image restored.","Referring to FIG.","9, a template 18 defines the watermark.","In the example of FIG.","9 the watermark is in the form of an S as in Sony but the watermark could have any other form.","The template defines the watermark to the resolution of a DCT block Templates are further discussed below in the section headed “Templates”.","Referring to FIG.","10, in overview, the first example of the invention may operate as follows.","These operations are performed only on I frames in this example but the invention is not limited to performing these operations on I frames.","In step S2, the macroblocks which fall within the watermark S are selected and all the DC differential and AC VLCs of those macroblocks are removed and in step S4 stored.","In step S6, new DC differentials which represent the watermark are put into the blocks of the template macroblocks in place of the ones which were removed.","(The AC VLCs are not replaced in those blocks).","The DC coefficients in I frames are differentially encoded and so the new DC differentials need to be calculated in dependence on the DC differentials of the macroblocks spatially preceding and following the macroblocks at the edges of the template as will be described in more detail below.","In step S8 the number of bits Nr removed from an I frame by removing the original DC differentials and AC VLCs is determined and the number of bits Na added to the I frame by the new DC differentials is determined.","The difference Nr-Na indicates the amount of spare data space in the picture data.","If that space exceeds a threshold amount (5 bytes for MPEG2) then referring to FIG.","11B a visible watermark user data space 20 is created in the picture data.","At least some of the stored DC differentials and AC VLCs are placed in that data space 20.To ensure that the bit rate of the picture data is not changed zero padding 22 may be added to the picture data.","If insufficient spare space is available to create the data space 20 then zero padding 22 is added.","The coefficients placed in the data space 20 are encrypted before they are placed in the space 20 to ensure that only authorised persons can reconstruct the original image.","That process is repeated (S10) for each I frame in a video sequence.","Referring to FIG.","14, a portion of a video sequence is shown.","FIG.","14 shows only I frames for ease of explanation.","The picture data frames are shown in simplified form for ease of explanation.","If the sequence comprises P and/or B frames, the I frames would be separated by such frames.","The first I frame P1 of the sequence includes watermark user data space 20 which contains some of the removed coefficients of frame P1.Frame P2 has a data space 20 and the other coefficients removed from frame P1 are in that data space.","In the example of FIG.","14, the user data space 20 of frame P2 has capacity to contain some of the removed coefficients of the frame P2.The others of the removed coefficients of frame P2 are in the user data space 20 of frame P3.Frame P4 is shown as having no spare capacity for a user data space 20 and some of the coefficients removed from frame P3 are in the user data space 20 of frame P5.Thus as illustrated by FIG.","14, the coefficients removed in serial order from the image data space of the frames of a video sequence are taken from store, encrypted and put, in the same serial order, into watermark user data spaces 20 created in the frames of picture data but without increasing the bit rate of the picture data.","The end of the video sequence is indicated by an end of sequence code 26.The user data spaces created in the video sequence may not provide sufficient spare capacity to contain all the removed coefficients.","Thus as shown in step S12 of FIG.","10, the remaining coefficients are taken from the store, encrypted and appended to the end of the video sequence after the end of sequence code 26 as shown at 28 in FIG.","14.The data appended to the end of the MPEG2 bitstream is part of the MPEG2 bitstream but because it is not prefaced by a start code, the appended data is ignored by an MPEG2 decoder.","Although the process has so far been described only with reference to I frames, the video sequence typically also includes P and B frames.","Some P and B frames may include zero padding.","If so the zero padding may be removed and used for a watermark user data block and the removed original coefficients may be placed in the user data block.","The zero padding is reinstated in the process of restoring the original bitstream.","That overview omits some details of operation which will be discussed in the following discussion.","End of Block and Other Codes As best shown in FIG.","13, the original data 30 (DC differentials and AC VLCs) removed from the image data 16 includes the original (or copies thereof) end of block codes EOB.","The data 32 (i.e.","the new DC differentials) added to the image data 16 includes new (or the original) end of block codes EOB.","In addition the newly created watermark user data spaces 20 include userdata start codes 34 which increase the number of bits in the picture data frames.","When determining the spare capacity available in the picture data frames once the original AC and DC differentials have been removed only the DC differentials and AC variable length codes are included since the EOB codes remain.","Flagging Watermark Userdata Blocks Referring to FIG.","12, it is necessary to be able to distinguish a watermark userdata block 20 inserted by the watermarking process from any user data block originally present.","For this purpose, the following technique is used.","Regardless of whether a picture frame contains an original userdata block 36, if there is space within the picture frame to include a watermark userdata block 20 the watermark userdata block 20 is the last userdata block in the picture frame.","See FIG.","12B.","The last bit 38 of the last byte of the watermark userdata is set to ‘1’ (i.e.","send a whole number of bytes up to the last byte in the userdata then only send 7-bits of watermark data followed by a single ‘1’ bit).","As shown in FIG.","12C if a picture contains one or more original userdata blocks 36 and there is no space within the picture to include a watermark userdata block then the last bit 40 of the last byte of the last block of userdata is stored in the watermark data and replaced by a ‘0’.","If a picture contains no original userdata blocks and there is no space within the picture to include a watermark userdata block then no operation is required.","Changing the last bit as described above is only an example.","The first bit could be changed instead.","Encryption of Watermark Data, FIG.","15 The watermark data as it appears in the watermarked MPEG-2 file must not contain MPEG start codes since they will terminate userdata areas prematurely or cause other undesirable decoder behaviour.","The nature of the codes used to represent DC differentials and AC VLCs is such that start codes cannot be accidentally generated within the displayed image data.","It should not therefore be possible to generate them in the watermark data either providing care is exercised when generating flag information.","However, the encryption of the watermark data may result in start codes being produced.","It is therefore necessary to check the encrypted data for start codes and where they occur revert to the unencrypted watermark data (this does not affect security since it is only possible to determine which bytes have been left unencrypted when decrypting with the correct key).","Where it has been necessary to revert to the unencrypted watermark data, a check must also be performed to ensure that this did not result in the generation of another start code across the encrypted/unencrypted data boundary.","Any suitable encryption algorithm may be used.","Currently the AES algorithm is preferred.","FIG.","15 shows an illustrative encryption process using an Exclusive OR (XOR) operation.","Because the XOR operation is used, the decryption process is identical to the encryption process.","In FIG.","15: “idx” is an index count indexing the watermark data bytes to be encrypted (or decrypted); “zero_count” is a count of the number of consecutive zeros in the bitstream; “ubyte” is a byte of unencrypted watermark data; “ebyte” is a byte of encrypted watermark data; “r” is a random number generated by the encryption algorithm and is different for each byte; “zero_count++” means increment zero_count; and “idx++” means increment idx.","In overview, the encryption process is monitored to ensure that an MPEG start code is not generated by chance.","An MPEG start code is for example 3 hexadecimal bytes 00 00 01.Such a code is not generated in the unencrypted watermark data but it is possible it would occur by chance in the encrypted data.","If such a code is generated in the encrypted data the unencrypted data is used instead.","The process of FIG.","15 checks for a sequence of zeros (step S57) or a sequence of zeros followed by a one (step S58) in which case it outputs (S60) the unencrypted data ubyte, otherwise it outputs (S59) the encrypted data ebyte.","Referring to FIG.","15, the encryption (or decryption) process starts at step S50 and proceeds to step S51 in which idx and zero_count are set to zero.","In step S52, idx is compared to the number num_bytes of bytes of watermark data to be encrypted (decrypted).","If all the bytes have been processed, the process ends at step S53, otherwise it proceeds to step S54 which gets the (next) watermark data byte ubyte to be processed and step S55 which gets a random number r for encrypting the data byte.","In step S56 the data byte is encrypted by performing ebyte=ubyteXORr.","Steps S57 and S58 check whether there is likely to be an MPEG start code in the resulting data stream.","Step S57 checks whether ubyte or ebyte is a sequence of zeros in which case step S59 outputs ubyte.","Step S58 checks whether there is a sequence of zeros (zero-count >1) followed by a one (ebyte=1).","If yes, ubyte is output S60, otherwise ebyte is output S59.At step S61, if ubyte is zero, then the zero_count is incremented, otherwise zero count is set to zero in step S62 and finally at step S64 idx is incremented and the process returns to step S52.Differential DC Recoding The DC coefficients for intra blocks are coded differentially throughout a slice.","The original differential value is stored and a new differential value calculated and used to replace it as described above.","This allows the change to be made to provide a desired, e.g.","fixed, luminance level and gives good definition of the template design.","Referring to FIG.","4, assume an I frame is being modified, and assume the central macroblock 10 is the macroblock in the template to contain watermark data and the left and right hand macroblocks are outside the template.","The DC differential 60 of the left hand block is outside the template and is not affected by the watermark.","DC differentials 61 to 64 are in the template.","DC differentials 61 to 64 are removed from the macroblock and stored.","DC differentials 61 to 64 are replaced by new differential values representing the watermark.","Assume, for ease of explanation that the watermark is an area of white level.","DC differential 61 is replaced by a new differential value representing the desired white level and DC differential 62 to 64 which all represent the same white level as DC differential 61 are thus set to zero.","DC differential 65 is recalculated.","Exiting the template involves replacing the original differential value (i.e.","of block 65) with a new one.","There is no need to store the original differential value (of block 65) as the process can be reversed in the washer because it can be calculated from the difference between: a) the value of block 65 calculated from the watermarked differential values converted to non-differential values; and b) the original (washed) non-differential value of block 64.FIG.","4 assumes a template coincides with a macroblock.","That is advantageous in that it minimises the number of entry and exit points and maximises the number of zero DC differentials.","However, a template may not coincide with a macroblock.","P and B Frames Intra macroblocks in P and B frames can be modified as described above.","Non-intra coded macroblocks can have the AC VLCs removed as described above but non-intra encoded macroblocks are predictively encoded and so the changes are less perceptible.","Motion vector watermarking can only be performed on P and B frames.","Chrominance The foregoing description assumes watermark data is provided only in luminance.","However macroblocks may also include DCT blocks of chrominance information.","Watermark data may be included in the chrominance DCT blocks in the same way as described for luminance.","Also the watermark data may be included only in chrominance DCT blocks and not in luminance blocks but that is not currently preferred.","Overview of Restoring Original Image Data (FIG.","16) Referring to FIG.","13C, there is shown an example of a watermarked bit stream (in a temporary picture buffer 74) but as it will be transferred to a main buffer 70 to replace the original bitstream for transmission to a decoder.","When received at the decoder, the watermarked image will be displayed unless the original image is restored.","FIG.","16 illustrates an example of an overview of a process according to the invention for restoring (washing) the original image.","In step S20, the watermarked bitstream is stored and parsed.","In step S22, the encrypted watermark userdata is extracted from the bitstream and stored.","Also, any data appended to the end of the bitstream is stored.","In step S24, the stored encrypted watermark user data is decrypted to restore the original DC differentials and the AC VLCs.","In step S26, restored original DC and AC VLCs replace the DC differentials which represented the watermark.","The resultant bitstream is an MPEG2 compressed bitstream but omitting watermarking data.","In step S28, the restored bitstream is decompressed in conventional MPEG2 manner.","Because all the original coefficients were retained as user data and/or appended to the end of the bitstream, the original bitstream is exactly restorable.","The resulting washed file is a bit for bit match to the original unwatermarked file.","An example of a process for restoring the original image data is described in more detail in the section “Watermark Wash Code”.","Illustrative Implementation (FIGS.","17, 18 and 19).","Referring to FIG.","17, an illustrative watermarking system is shown An uncompressed digital video sequence from a source 50 is applied to a conventional MPEG2 encoder 52.Such encoders are well known.","In this example the encoder 52 is a computer running compression software but the encoder could be a special purpose processor.","The compressed video sequence is stored as a file in a store 54, e.g.","the hard disk of the computer 52.A watermark embedder 56 receives the compressed video sequence from the store 54.In this example the embedder is a computer running embedding software which will be described below but the embedder could be a special purpose processor.","The embedding process involves a template such as is shown in FIG.","9 and an encryption algorithm which uses at least one encryption key, both of which are needed to restore the original image at the decoder.","In this example, the computer has a smart card interface 57 via which the template and the key(s) are stored securely on a smart card 64 or other secure data carrier.","Alternatively, the template and key(s) may be transmitted in secure manner across a network or in any other secure manner.","The embedder applies the watermark to the compressed bitstream and the watermarked bitstream is transferred to another file store 60 (for example in a studio 62) via a transmission link 58 which may be a wireless link, a network, a data storage device for example a disc or tape, amongst other examples.","Referring to FIG.","18, a decoder is shown.","The watermarked bitstream is taken from the store 60 and applied to a de-embedder also called a “washer” 66 which restores the original bitstream.","The washer in this example is a computer running de-embedding software which will be described below but the washer could be a special purpose processor.","The washing process involves the template such as is shown in FIG.","9 and the encryption algorithm which uses at least one encryption key, both of which are needed to restore the original image at the washer.","In this example, the computer has a smart card interface 63 via which the template and the key(s) are received from the smart card 64.The output of the washer is a compressed bitstream which is fed to a conventional MPEG2 decoder 65 for decompression and display on a display 67.Buffers Referring to FIGS.","19 (and 13), for the purposes of embedding the watermark data in the compressed bitstream, a file (54 in FIG.","12) and three buffers are used.","They are: a main buffer 70 which stores the original MPEG2 compliant compressed bitstream from the compressor and which outputs the watermarked bitstream; a watermark data buffer 72 which stores the original DC differentials and AC VLCs; and a temporary picture buffer 74 in which the watermarked bitstream is constructed for transfer to the main buffer.","These buffers may be physical buffers or in this example are virtual buffers defined by software.","The washing process correspondingly uses three buffers, a main buffer 70, a watermark data buffer 72 and a temporary buffer 74.Watermark Embedding The following sets out, by way of example, a pseudo code implementation of various processes described above.","Watermark Embed—Pseudo Code Implementation Example Create a main buffer and a watermark data buffer WHILE pictures still to do Read data for one picture from file into the main buffer Create a temporary picture buffer WHILE NOT at start of displayed image data Parse the main buffer Enable all selected watermarking techniques (flag in the watermark data buffer) WHILE NOT end of picture CALL Process Slice Check that space constraints can be met IF space constraints not met Disable some of the watermarking techniques for this picture (flag in the watermark data buffer) WHILE NOT end of picture CALL Process Slice Determine space available in picture with new watermarked displayed image data IF less than 5 bytes IF original user data present Flag the last block of userdata to indicate it is not watermark data Output the displayed picture data to the main buffer Pad the picture to size using zero bytes ELSE Output a user data start code WHILE there is watermark data AND there is space in the picture to fill Encrypt watermark data Output encrypted watermark data to user data space in the temporary picture buffer Flag the block of user data to indicate it is watermark data Output the displayed picture data to the main buffer IF there is space in the picture to fill Pad the picture to size using zero bytes Write the modified picture from the main buffer to file Write any data remaining in the watermark data buffer to file Process Slice function WHILE in slice IF data is not related to the watermark techniques POP the data from the main buffer PUSH the data to the picture buffer ELSE POP the data from the main buffer PUSH the data to the watermark data buffer Calculate replacement value Encode replacement value PUSH replacement value to the picture buffer Watermark Embed Code By way of explanation of the Watermark Embed code, the MPEG2 compliant bitstream is stored in a file, e.g.","54 in FIG.","12, the buffers 70, 72 and 74 are created and the data for one picture is written from the file into the main buffer 70 where it is parsed.","Next selection of watermarking techniques takes place.","The selection of watermarking techniques” are described in more detail below with reference to FIG.","22.Initially, all watermarking techniques are enabled.","The “process slice function” which will be described in more detail separately is then run.","In overview, that function places the original DC differentials and AC VLCs in the watermark data buffer 72; and calculates the new watermark DC differentials (i.e.","replacement values) and places the replacement values in the picture buffer 74.A check is then made to ensure that a space constraint can be met.","Initially it is assumed that all of a particular set of watermarking techniques is applied to the bitstream.","If that does not satisfy the space constraint, then one or more of the techniques are disabled.","One space constraint, in this example, is passed if the number of bits freed by removing AC VLCs from the image is greater than the number of bits needed to make changes to the DC differentials.","That indicates that the watermarked display image is no greater than the original display image.","If it is not met then that indicates that the removal of the AC VLCs has not freed enough space for the watermarking DC differentials.","The process slice function is run again with the selected techniques.","A step “Determine space available”, in this example, checks the space available in the picture with the new watermark DC differentials in it.","If there is less than 5 bytes of spare space then no watermark user data field 20 can be included.","If the bitstream includes one or more user data fields 36, the last bit of the last such field 36 is set to zero as shown at 40 in FIG.","11C.","The displayed picture data which includes the new DC differentials representing the watermark is output from the picture buffer 74 to the main buffer 70 and the picture size is padded to its original size by zero bits; see 22 in FIG.","11 or 12.If for example, there is sufficient space in the picture data including the new DC differentials representing the watermark, then a watermark user data field 20 is created in the picture buffer 74 by outputting a user data start code.","The original AC VLCs and DC differentials stored in the watermark data buffer 72 are encrypted and placed in the watermark user data field in the temporary picture buffer 74.The last bit of the watermark user data field is set to 1 as shown at 38 in FIG.","11B to indicate it contains the original AC VLCs and DC differentials.","The contents of the temporary picture buffer 74 are output to the main buffer 70.If there is space to fill in the watermarked displayed image data space 23, the space 23 is filled with zero bits as indicated at 22 in FIG.","11C.","The data in the main buffer 70 is then written to a file.","Any data remaining in the watermark data buffer 72, i.e.","the original AC VLCs and DC differentials which are not in the watermark user data, are encrypted and also appended to the end of the bitstream forming the file.","Process Slice Function.","By way of explanation of this function, data is processed, in this example, in slices as described with reference to FIG.","5 above.","However that is not essential.","It is merely a convenient way for the software to operate.","The code POP indicates that data is removed from a buffer.","The code PUSH indicates that data is moved into a buffer.","Thus the data in the main buffer 70 has been parsed as described above.","All data up to the start of the displayed image data is skipped over in the main buffer and left there.","The only data which is PUSHed into the picture buffer is the displayed image data.","This makes it easier to modify an original user data block.","In cases where watermark user data is to be added, it is necessary to insert this in front of the watermarked display image data.","Data which is related to watermark data, e.g.","the AC VLCs and DC differentials are removed from the main buffer 70 and put into the watermark data buffer 72.The new replacement values of the DC differentials are calculated and encoded and put into the picture buffer 74.Watermark Wash—Pseudo Code Implementation Example Create a main buffer and a watermark data buffer CALL Initialise Watermark Data WHILE pictures still to do Read data for one picture from file into the main buffer Create a temporary picture buffer CALL Get Watermark Data WHILE NOT at start of displayed image data Parse the main buffer Enable watermarking techniques as indicated by the setting of flags in the watermark data WHILE NOT end of picture CALL Process Slice Output the contents of the picture buffer to the main buffer IF there is space in the picture to fill Pad with zeros Write the modified picture from the main buffer to file Initialise Watermark Data Function Isolate watermark user data blocks from the file Decrypt the data Store in a temporary file Get Watermark Data Function IF watermark data buffer is not full AND watermark data still present in temporary file Read watermark data from temporary file into watermark data buffer Process Slice Function (not the Same as the Function Used to Embed the Watermark) WHILE in slice IF data is not related to the watermark techniques POP the data from the main buffer PUSH the data to the picture buffer ELSE POP the data from the main buffer POP original data from the watermark data buffer PUSH original value to the picture buffer Watermark Wash Code The watermark wash code restores the original image data.","By way of explanation, a main buffer 70 and a watermark data buffer 72 and a temporary buffer 74 are created at the decoder 66.Also a temporary file is created by a function “Initialise Watermark Data function”.","The initialise watermark data function isolates the watermark user data blocks from the file, i.e.","the encrypted original AC and DC differentials, decrypts them (using the key from the smart card 64) and stores the decrypted AC VLCs and DC differentials in the temporary file.","If there is picture data which has not been washed, that data is read from the temporary file into the main buffer.","The temporary buffer 74 is created at this stage.","A function Get Watermark Data determines whether the watermark data buffer 72 is not full AND watermark data is still present in the temporary file.","If so, then watermark data is read from the temporary file into the watermark data buffer.","This ensures that there is always at least enough data in the watermark data buffer to wash the current picture.","“Enable watermarking techniques .",".",".","” reads the 1 bit flag from the watermark data to determine if the picture was successfully watermarked using all selected techniques.","If it was not, then additional flag bits are read to determine which techniques were used.","CALL process slice operates as follows: POP and PUSH indicate the same operations as given above.","Data in the main buffer has been parsed.","Data in the main buffer which is not related to watermarking is removed and placed in the picture buffer 74.Watermark data (i.e.","the DC differentials which represent the watermark) is removed from the main buffer 70.The decrypted original AC VLCs and DC differentials in the watermark data buffer 72 are removed therefrom and put into the picture buffer 74.Further steps transfer the data from the picture buffer to the main buffer Finally, the restored picture is written from the main buffer to a file.","It will be appreciated that the process described above requires the template stored in the data carrier 64 to determine which blocks contain watermark data and which do not.","Also the key(s) needed for decryption are derived from the data carrier 64.Motion Vectors which have been modified as described in the section below “Fourth Example—Motion Vectors” are also restored to their original values in the same way.","SECOND EXAMPLE Non-Intra DC Changes In the first example, the watermark is applied by replacing the original DC differential values of the DC coefficients in template blocks in Intra frames with new values representing the watermark.","No other coefficients are replaced.","The original values are stored in the watermark user data pspace and/or at the end of the bitstream.","In a second example of the invention, in addition to replacing the intra frame DC coefficients, the DC coefficients in template blocks in non-intra frames (that is P and B frames) are replaced by new values and the original values stored in the watermark user data space and/or at the end of the bitstream.","In non-intra frames, DC coefficients are coded as the difference between the current DC coefficient and the desired coefficient is coded since the magnitude of the change is usually small.","However, there are instances where the magnitude might be much larger and this may cause the DC value of a watermarked block to clip at the limits of the range.","To avoid this the magnitude of the DC coefficient can be changed to the minimum value of 1.","(Changing the value to zero would completely eliminate the VLC and alter the run-length of the following code).","The original VLC is stored in the watermark data buffer 72 and the new code put in its place.","The VLC representing a run-length of zero and a level change of ±1 is the shortest VLC in the coding table so this process guarantees never to increase the number of bits required to code the picture.","It can therefore always be performed whenever intra DC changes are also being performed.","The sign of the level change is set to be positive when the DC change is to black and negative when the change is to white.","This prevents clipping of the DC from occurring.","Referring to FIG.","20, non-intra frames are processed as follows.","Step S30 determines the frame type.","If the frame is a non-intra frame the process proceeds to step S32, otherwise if the frame is an intra-frame the process proceeds as set out in FIG.","10.In step S34, the codes representing DC coefficients (which are difference values as described above) in template blocks are removed and stored in the watermark data buffer 72 (see FIG.","12B).","In step S36, new change values of +/−1 are placed in the bitstream in place of the removed coefficients.","In steps S36 and S38, the bitstream is reformed in the temporary buffer 74 (see FIG.","12C), the removed coefficient values being placed in the watermark userdata space 20 if space is available and otherwise in step S40 appended to the end of the bitstream.","The original DC coefficients are encrypted as described above.","AC VLCs are not changed in non-intra frames.","THIRD EXAMPLE Frame Pictures, Changing DCT type, FIG.","21 The first and second examples may be modified as follows.","There are two types of picture: field and frame.","In field pictures only one type of DCT block occurs.","In this, third, example, we are concerned only with frame pictures.","In frame pictures two types of DCT blocks can occur; field and frame.","The field DCT is used when the video source is interlaced because it improves the correlation between adjacent rows and consequently increases the proportion of the signal power in the lower frequency components of the DCT.","However, it is not used exclusively for this purpose.","In a macroblock all the blocks have the same type.","Referring to FIG.","21A, if a DC change is made in a frame picture to one DCT block B1 but not to the DCT block either directly vertically above or below it in the same macroblock MB and the field type is being used for the DCT blocks of that macroblock then the change will result in stripes across the entire height of the macroblock.","This is not desirable as it distorts the watermark and if the watermark is in the form of alpha-numerics they may become harder to read.","The appearance of stripes is entirely dependent on the encoding of the content and in particular the proportion of macroblocks coded with the field DCT.","In accordance with an example of the present invention, to solve this problem the DCT type of any macroblock in a frame picture, which contains at least one DCT block to be watermarked, is set to frame.","This involves storing the original DCT type code in the watermark data buffer 72 (FIG.","13) and replacing it with the code for a frame DCT.","This does not to affect the number of bits required by the picture since the DCT type code is always a 1-bit value.","This change can therefore always be made whenever the DC watermark is used.","As with motion vector changes, the DCT type operates at the macroblock level so any change will affect all DCT blocks within the macroblock regardless of whether they are watermarked or not.","In this case the side-effect that occurs is a re-ordering of the rows within the macroblock.","This may result in an obviously, visibly, incorrect macroblock but the appearance of the macroblock may not be objectionable.","FIG.","21B illustrates an example method to process the DCT type (including, where appropriate, changing from field to frame DCT and reversing this change).","In FIG.","21B: “picture_structure==FRAME” detects Frame pictures; “&&” is logical AND “frame_pred_frame_dct ==0” detects that the macroblocks have the DCT type specified on an individual basis rather than being universally set to frame DCT type for this picture; ∥ is logical OR; and “macroblock intra∥macroblock_pattern” detects intra-encoded macroblocks and non-intra blocks which include coded DCT blocks.","Intra-encoded macroblocks have all DCT blocks coded.","It is necessary to detect this because only coded blocks have a DCT type.","DCT type (dct_type) is denoted by a 1 bit code: 1=field type; 0=frame type.","The original bitstream is stored in a main buffer 70 and the codes which are removed are temporarily stored in a watermark data buffer 72 (see FIG.","13).","The process starts at step S70 and proceeds to step S71 and S72 which detect macroblocks which have a DCT type.","If a macroblock has no DCT type the process ends at step END.","If a macroblock has a DCT type then the 1 bit code representing DCT type is removed (POPped) from the main buffer 70 in step S73.Step S74 determines that DC changes are enabled (see the section “selective techniques” below) and the macroblock is within the template.","If yes, step S75 determines whether a watermark is being embedded in the bitstream or a watermarked bitstream is being restored to its original form (washed).","If the bitstream is being watermarked, then in step S76, the dct_type code (that was removed from the main buffer in S73) is placed in (PUSHed into) the watermark data buffer 72 (see FIG.","13).","The value of the dct_type code is then changed to frame type (=0) in step S77 and in step S79 the new code is PUSHed into the temporary buffer 74 (see FIG.","13).","If the NO route is taken from step S74 the POPped dct_type code is PUSHed into the temporary buffer in step S79.In the reverse process steps S70 to S74 proceed as described above.","Then at step S75 it is determined that the watermarking process is being reversed (washing) and at step S78 the dct_type code for the watermarked stream is replaced by the original one that is extracted from the watermark data buffer 72.In step S79, the dct_type code is then PUSHed into the temporary buffer 74 as before.","At the end of the process shown in FIG.","21B, the temporary buffer 74 contains the new dct_type code and the watermark data buffer 72 contains the original codes.","The original dct_type codes from buffer 72 are transferred to the watermark user data spaces 20 or appended to the end of the bitstream.","The new dct_type codes are placed in the bitstream in place of the original codes.","That is done in the same way as described above in the section “Watermark Embed Code”.","Selective Techniques As described above, a picture is watermarked by: a) First Example—removing all codes representing original DC and AC coefficients from template blocks in intra frames and replacing the DC coefficients by new values representing a watermark; b) Second Example—optionally, in addition removing codes representing original DC coefficients in non-intra frames and replacing them with new values; c) Optionally, in addition, replacing original motion vector values of template blocks with zero values.","More generally three techniques may be used: 1.Enabling change of DC coefficients in template blocks; 2.Enabling removal of AC coefficients from template blocks; and; 3.Enabling change of Motion Vector values in template blocks.","Usually, a picture watermarked using the example techniques described here will require fewer bits to encode than the original picture.","In the first example this is due to the complete removal of AC coefficients but the coding of a large number of zero differential values for both DC and motion vectors also makes a significant contribution.","However, there are instances where, to embed the watermark completely, the number of bits inserted into a picture ends up being greater than the number of bits removed from that picture.","For example, this can easily occur with a completely black frame, commonly encountered in video sequences.","If this occurs, then in order to satisfy the constraint that the number of bits used to represent the picture must remain the same it becomes necessary to disable some of the techniques used to watermark the picture until the constraint can be met.","In examples of the invention, the selection is made on a per-picture basis so it is necessary to store additional information to facilitate correct washing of the picture.","Flag bits are inserted to indicate which techniques are enabled for the current picture.","The system used at present is shown in Table A.","It easily adapts to the addition of further techniques and is also efficient regardless of the selected subset of techniques chosen from the full set of techniques available.","Note that the choice of the first flag bit is designed to prevent the occurrence of MPEG-2 start codes within the watermarked data.","TABLE A Selective technique flag bits Whenever possible the intra DC change and AC removal techniques should both be applied as a minimum since this gives good visibility and robustness.","If intra DC changes are applied then non-intra DC and DCT type changes can also be used since they are guaranteed not to increase the number of bits required by the picture.","Changes to the motion vectors improve the visual quality and integrity of the watermark but are not essential.","FIG.","22 is a flow diagram illustrating the selection, by way of example, of the three watermarking techniques: 1.Enabling change of DC coefficients in template blocks; 2.Enabling removal of AC coefficients from template blocks; and; 3.Enabling change of Motion Vector values in template blocks.","The following points should be noted: AC bits freed is always non-negative.","AC removal can always be performed and will never result in an increase in picture size.","Bits for DC changes and Bits for MV changes may be positive or negative.","If the value is positive then the number of bits needed to perform the technique is greater than the number of bits originally present.","A sufficient number of bits must therefore be freed using another technique (e.g.","AC removal) if the changes are to be performed on the current picture.","If the value is negative the technique requires fewer bits than originally present so some bits have actually been freed and the changes can always be performed.","DC changes are given priority over MV changes whenever the number of bits freed is insufficient to compensate for the bits needed to perform both techniques Referring to FIG.","22, the process starts at step S80 and proceeds to step S81 at which initially all three techniques are enabled.","In step S82 a picture (that is a frame or field) is processed to apply all of the three enabled techniques.","Step S83 sets a reprocess flag to true to indicate that initially it is assumed that the picture can be reprocessed using all three techniques.","In overview, a picture is processed in one pass using all three techniques to apply a watermark and the result is analysed.","If the result is not successful, the picture is reprocessed in a second pass with one or more of the techniques disabled.","AC Bits Freed Allowing All Three Techniques Step S84 determines whether removing the AC coefficients removes any data thus freeing bits for changing the DC coefficients.","(For a black picture there may be no AC coefficients).","If the number of bits freed exceeds zero then step S85 determines whether the number of AC bits freed exceeds the sum of bits for DC changes and bits for MV (Motion Vector) changes.","The number of bits for DC changes is the difference between the number of bits freed by removing the original DC codes and the number of bits used by the new DC codes.","Likewise the number of bits for MV changes is the difference between the number of bits freed by removing the original MV codes and the number of bits used by the new MV codes.","If the number of AC bits freed is NOT less than the sum of the bits for DC changes ad the bits for MV changes (i.e.","sufficient AC bits are freed to allow both DC and MV changes) then all three techniques can be used and so there is no requirement to reprocess the picture.","Thus the reprocess flag is changed to false in step S90.Step S91 determines whether the reprocess flag is true or false.","In this instance it is false so the process ends at the END step.","Step S84—no AC Bits Freed This may occur on a black picture which thus has no AC coefficients and so it is not possible to embed any watermark.","That is not a problem because a black picture has no content to protect.","If in step S84 it is determined that no AC bits are freed then the process proceeds to step S93 which disables the AC removal technique.","Step S94 tests whether the bits for DC change is greater than zero.","If it is then the DC change technique is disabled in step S95.If there are no bits for DC change or step S95 has occurred the process proceeds to step S96 which test whether the number of bits for MV change is greater than zero.","If it is then in step S97 the MV change technique is disabled.","In summary if no AC bits are freed, the DC and MV changes are disabled or there are no DC and MV change bits.","In either case the reprocess flag is still true and so the picture is processed (again) at step S92.Note that whilst ‘AC bits freed ’ is always non-negative, ‘bits for DC changes’ and ‘Bits for MV changes’ may be positive or negative.","If there is no AC data to remove, then DC and MV changes can only be performed if the replacement data has fewer bits than the original data: i.e.","‘Bits for DC changes <=0 and similarly for MV changes.","It is therefore possible (but unlikely) to have DC and/or MV changes enabled with AC removal disabled.","It is always necessary to reprocess when any of the techniques is disabled since the original assumption is that all three techniques are enabled so the flag data must be changed and the first pass watermarked frame is overwritten by the second pass watermarked frame.","The lack of AC VLC data means that no space can be freed in the picture data to allow changes in the differential DC values that increase the number of bits.","In such instances it may not be possible to embed the watermark but this is not a problem since there is no content to protect.","However, a decoder which removes the watermark herein referred to as a watermark washer must be told where these instances have occurred so that it does not attempt to remove the watermark from the frame.","Additional flag bits are used to indicate this.","AC Bits Less than the Sum of Bits for DC and MV Changes Step S85.If there are insufficient bits freed to do both DC and MV changes then the process proceeds to step S86.In this case, step S86 tests whether the number of AC bits freed is sufficient to do the DC changes.","If there are sufficient bits to do the DC changes (NO output of S86) then step S97 occurs and MV changes are disabled.","The process then proceeds to step S91.Because the reprocess flag is true, but MV changes are disabled, then the picture is processed anew with MV changes disabled and DC changes enabled in step S92.If step S86 determines that there are insufficient bits freed to do the DC changes (YES output of S86), then the process proceeds to step S87 which tests whether the number of AC bits freed is sufficient to do the MV changes.","If there are sufficient bits to do the MV changes (NO output of S87) then step S88 disables DC changes.","The process then proceeds to step S91.Because the reprocess flag is true, but DC changes are disabled, then the picture is processed anew with DC changes disabled and MV changes enabled in step S92.If step S87 determines that there are insufficient bits to do the MV changes (YES output of S87) then step S890 disables both DC and MV changes and the picture is reprocessed via steps S91 and S92 with only the AC removal technique enabled.","In a modification of the process of FIG.","22, AC removal is permanently enabled because it has been found that AC removal never causes problems.","In FIG.","22, step S93 is then removed and a fig bit for AC removal is not required.","FOURTH EXAMPLE Motion Vectors—FIGS.","23 to 25 Setting Motion Vectors to Zero Referring to FIG.","23A, motion vectors are associated with macroblocks (shown by bold squares MB) rather than DCT blocks (shown by lighter squares B).","Thus a motion vector template as indicated by light shading may be defined based on the watermark template shown by dark shading.","Even if, as assumed above in the first example, only I frames contain watermark data, motion vectors may undesirably distort the watermark.","It is preferable but not essential to determine whether any macroblock in a P and or B frame falls within the motion vector template.","If it does, then the value of any motion vector of that macroblock is set to zero.","Motion vectors are differentially encoded so the differential value is set to represent a value of zero for the motion vector.","(That is, Motion Vectors are processed in the same way as described above for the DC differentials).","As described above with reference to the DC differentials, the original codes representing the Motion Vectors of DCT macroblocks, which include DCT blocks within the template, are removed from the bitstream and replaced by modified codes which in this case represent Motion Vectors of zero value.","The original Motion Vector codes are placed in the watermark user data spaces and/or appended to the end of the bitstream.","Thus referring to FIGS.","13 and 24:— The bitstream is stored in the main buffer 70.In step S100, it is determined whether in a non-intra frame a macroblock falls within the motion vector template.","If it does then the motion vector code associated with that macroblock is removed from the bitstream (S102) in the main buffer 70 and stored (S104) in the watermark data buffer 72.A new motion vector code representing zero value for motion vector is placed in the bitstream in place of the original value in the temporary buffer 74 (S106) and the original motion vector codes are placed in the watermark user data space 20 in the temporary buffer 74 (step S108) and/or appended to the end of the MPEG compliant bitstream (S110).","The original motion vector codes are encrypted before being placed in the user data space or appended to the end of the bitstream as described above with reference to FIG.","15.Motion Vectors may be set to zero as described in this section in any of the examples given above.","Whilst this reduces distortion of the watermark, the watermark may still be distorted especially at the edges.","The next section headed “Borders” describes an illustrative method of further reducing distortion of the watermark.","Borders FIG.","23B Referring to FIG.","23B, to further reduce distortion of the watermark, the motion vector template is extended to include macroblocks which do not include watermark template blocks but which border such blocks.","The motion vector codes of all the macroblocks in the extended motion vector template are processed as described above with reference to FIG.","24.This reduces the distortion of the watermark as defined by the watermark template but because the motion vectors of the macroblocks bordering the watermark template, which do not include any watermark blocks, the picture is distorted in the bordering macroblocks.","It is a matter of subjective judgement whether the reduced distortion of the watermark is outweighed by the increased distortion of the picture.","Changing the Values of Motion Vector Codes, FIG.","25 Motion Vectors are differentially encoded, and are recoded, in the same way as DC coefficients as described above in the section headed “Differential DC recoding”.","P frames have forward prediction motion vectors but B frames have two types of motion vectors: forward prediction and backward prediction.","The two types are processed separately as will be described with reference to FIG.","25.FIG.","25A shows a much simplified motion vector template of three macroblocks MB between other macroblocks O which are not part of the watermark and its border.","FIG.","25B shows the prediction types of the macroblocks.","F indicates forward and B indicates backward.","Some macroblocks FB are both forward and backward predictive.","Referring to FIGS.","25 C to E, the numbers shown in those Figures are for explanation only and do not purport to represent realistic values for motion vectors.","FIG.","25C shows non-differential values for forward predictive motion vectors.","The corresponding differential values are shown in FIG.","25D.","It will be noted that the backward predictive motion vectors are ignored.","When the motion vectors in the three template macroblocks are set to zero, the differentials of those blocks are recalculated so the effective values within the template are effectively zero but the effective values of neighbouring blocks outside the template are not changed.","That results in the differential value of the macroblock at the leading edge of the template denoted “Entry” changing relative to its preceding macroblock to provide effectively zero motion vector value and the differential value of the trailing edge macroblock denoted “Exit” changing to maintain the original value of the next bordering macroblock.","It will be appreciated that the forward predictive motion vectors values are in a sequence of differential values which are independent of the backward predictive values and that the Entry and Exit macroblocks of the two sequences do not necessarily coincide.","It will also be appreciated that the original differential value of the motion vector of the macroblock bordering the exit from the templates does not need to be stored as described above for block 65 in the section “Differential DC Recoding” but it is preferred to store it.","Templates—FIGS.","26 to 28 As discussed above, there are two types of picture: field and frame.","A video sequence may contain: only field pictures; only frame pictures; or both field and frame pictures.","Sequence of Frame Pictures Only Consider a sequence of only frame pictures.","A template is defined to a resolution of one DCT block horizontally and vertically (8 pixels by 8 pixels).","For the example frame picture shown in FIG.","26A, having 720 pixels per line and 576 lines, the required template size is 90 blocks by 72 blocks.","A template in the form of an X is defined to the resolution of one block in FIG.","26B.","The resulting watermarked picture shown in FIG.","26C has a displayed watermark resolution of one block of 8×8 pixels.","Sequence of Field Pictures Only Consider a sequence of only field pictures.","Referring to FIGS.","27 A1 and A2, two fields each of 720×288 pixels are interlaced to define an interlaced frame of 720×576 pixels.","In a field picture, a DCT block still corresponds to an 8×8 pixel block even though the number of lines is half that of a frame picture.","A macroblock is 2×2 DCT blocks as in a frame picture.","A template (X in FIG.","27B) for a field picture is defined to the resolution of one DCT block.","The same template is used for both fields to prevent the occurrence of stripes.","No special adjustment of the template for each field is required because block positions are fixed relative to the start of a field.","Referring to FIG.","28 in respect of one macroblock as shown in FIG.","28C a template indicates one block shown in solid black is to be watermarked.","In the interleaved frame, two macroblocks from the respective two fields are interleaved to form a composite of two DCT blocks horizontally by four DCT blocks vertically as shown in FIGS.","28A and 28D.","The composite consists of a macroblock B1 from field one and a macroblock B2 from field two.","The lines of the interlaced frame are indicated by F1 for lines from field one and F2 for lines from field two.","The watermark X as displayed has a resolution of 8 pixels horizontally by 16 pixels vertically as shown in FIG.","28D even though the template has a resolution of 8 pixels by 8 pixels as shown in FIG.","28C.","Sequence of Field and Frame Pictures Consider a video sequence having at least one field picture and at least one frame picture.","The template, X in FIG.","27B, is designed for the one or more field pictures and has a resolution of one DCT block.","That results in a displayed watermark having a resolution of one block horizontally by two blocks vertically.","The watermark in the one or more frame pictures needs to be consistent with that of the field pictures; this can be ensured by automatically deriving the template for frame pictures from the template used for field pictures.","The simplest way to achieve this is to double up every row of the field template.","The frame template then has an effective resolution of one block horizontally by two blocks vertically to ensure the displayed watermark in the frame pictures matches that of the field pictures.","In practice a video sequence is examined to determine the type(s) of picture in it.","For field-only and frame-only sequences, a suitable template for that type of picture is used.","For a sequence having both types of picture, a template suitable for field pictures is chosen and used for the field pictures in the sequence.","A template for use with the frame pictures is then derived from the field template to ensure a consistent appearance to the watermark.","The templates may be bit-maps stored in one or more stores.","Modifications Packetised Bit Stream Where the original sequence is an MPEG-2 packetised stream (either packetised video sequence or audio/video multiplex) the video sequence end code is not required provided that the watermarked video sequence is repacketised so that the residual watermark data is placed after the Packetised Stream end code.","To simplify the embedding of packetised streams (audio/video multiplex), the stream can be pre-processed to extract the video sequence.","The video sequence can then be embedded separately as described above.","The embedded video sequence can then be repacketised with the audio using the existing packetised stream file as a template, allowing reuse of all packet information and preserving all timing (a useful side-effect of maintaining the picture data size).","This additional stage of processing is not described in the psuedo code above since it is not essential to the operation of the embedding process.","Varying Compression Ratio The examples described above assume the video is compressed for minimum compression to maximise picture quality before the watermark data is embedded in the compressed bitstream.","Thus the watermark user data fields are unlikely to contain all the encrypted original coefficients and the residual data is appended to the end of the file.","In a version of the invention, which is not currently preferred, an estimate of the data space needed to contain the watermarked data is provided before compression is carried out and the compression controlled to provide sufficient spare space to accommodate all the original coefficients in watermark user data fields and the new DC differentials in the displayed image data fields.","Appending the Encrypted Original AC VLCs and DC Differentials.","In the examples described above the original AC VLCs and DC differentials in excess of those stored in the watermark user data fields are appended to the end of the file.","However, they could be placed at the beginning of the file.","In a currently preferred implementation the number of residual bytes appended to the end of the file is counted.","The count is placed after the residual data to enable the start of the residual data to be easily accessed.","Alternative Method of Storing Original AC VLCs and DC Differentials in User Data Fields In the foregoing description, the original AC VLCs and DC differentials are stored in the user data fields 20 of the compressed bitstream.","As shown in FIG.","14, in the example of frames P1 and P2, some data of frame P1 is stored in the user data space 20 of frame P1 and any data in excess of that is stored in the user data space of frame P2.That process continues throughout the bitstream.","Any residual data is appended to the end of the bitstream.","In a modification of that process, as shown in FIG.","29, original AC VLCs and DC differentials of frame P1 are stored in the user data space 20 of frame P1.For every frame there is produced a residual data field 291 comprising the number of bytes B of encrypted AC VLCs and DC differentials which are not included in the user data space of that frame together with the encrypted AC VLCs and DC differentials.","If there are no residual AC VLCs and DC differentials, the number B for that frame is zero.","Thus each frame has in the user data spaces 20 thereof only data originating from that frame and the residual data is stored elsewhere for example in a buffer 290 in FIG.","29.The residual data, i.e.","contents of the buffer 290 in this example, may be appended to an end of the MPEG bitstream as shown in FIG.","30.As indicated at 292 it may be appended to the end at the start of the bitstream or as indicated at 293 it may be appended to the end at the finish of the bitstream.","The codes B representing the number of bits of user data are chosen so that they do not represent or can be confused with other codes used in MPEG bitstreams, especially MPEG start codes.","Files As described above, any residual data comprising the codes B and the encrypted residual AC VLCs and DC differentials is appended to an end of the bitstream.","In a modification, that residual data is stored in a file separate from the bitstream.","The file may be appended to an end of the bitstream as shown in FIG.","30 at the beginning 292 of the bitstream or at the finish 293.As shown in FIG.","31, the file may be stored in a transmission medium 294 for example a data carrier or a communications channel separate from the bitstream.","The data carrier may be a floppy disc or tape or other storage medium.","The file is associated with the bitstream.","That may be done by appending it to the bitstream as shown in FIG.","30.If the file is stored separately as shown in FIG.","31 it may be associated with the bitstream by use of for example an identifier which links the file to the bitstream.","An example of such an identifier is a UMID.","Washing To wash the watermark from the bitstream of FIG.","29, the bitstream P1 to Pn is stored and parsed.","The residual data comprising the codes B and the AC VLCs and DC differentials is retrieved from the appended data 292 or 293 in the parsing process or from the file 292, 293 or 294 and stored.","For each frame, the data in the user data field 20 is decrypted and used to replace the modification data in the bitstream.","The number of bytes B is read for that field.","If it is greater than zero, then the residual data for that field is decrypted and used to replace the remaining modification data of that frame.","In the example of an MPEG bitstream the frames are not numbered nor are the residual data fields 291.Such numbering is not needed: every frame and its associated residual data field are processed in succession.","Such numbering could be provided in other examples of the invention.","Selecting Watermarking Techniques The foregoing description describes the selection of three watermarking techniques.","However selection could be made from many techniques including: a) replacement of DC differentials by modified DC differentials and removal of the original DC differentials; b) removal of AC VLCs to make space for the original DC differentials in the bitstream; c) placing removed coefficients in user data blocks; d) placing removed coefficients at the end of the bitstream; e) modification of motion vectors for watermarking; f) modification of colour information for watermarking; and g) placing coefficients in spare space in P and/or B frames.","The techniques may be used separately or together.","For example it may be possible in some circumstances to replace only the DC differentials with modified coefficients without removing AC VLCs.","Typically, but not always, modification of coefficients and motion vectors for watermarking increases the number of bits.","The techniques used may be selected depending on the effect they have on the bitrate.","If a particular set of techniques results in too many bits (e.g.","a) plus c)) then a different set of techniques may be selected.","The techniques used are indicated in the watermarked bit stream by flags which identify the techniques so that when the watermarked bitstream is applied to a washer for removing the watermark the washer is supplied with the information needed to correctly remove the watermark.","Applications The examples of the invention described above can be applied in the following contexts.","Content Brokering The visibly watermarked files produced using the technique described allow preview of content using a standard MPEG-2 player (the watermarked file is a valid MPEG-2 stream) there is no need for a bespoke player.","The watermarked file contains all information needed to reverse the process (apart from the encryption key and the template file).","It is therefore only necessary to distribute/transmit one copy of the content (as compared with the conventional procedure of distributing a protected copy of the sequence followed by an unprotected copy when the buyer has purchased the content).","There are a number of forms that this distribution can take: The watermarked content could be stored on a secure server but the encryption of the watermark data within the file could be performed at the moment a copy of the file is requested.","The client can enter their details which are then stored on the secure server with a copy of the encryption key used.","If the client then wishes to purchase the content the encryption key can be retrieved from the secure server using the client's details as an index.","This means that one client's key cannot be used to wash another client's copy of the watermarked content.","Of course, it would still be possible for the client with the key to provide their copy of the content (either watermarked or washed) to someone else.","Once the client has purchased the content there is the option of either washing the file to produce a file without the visible watermark or to leave the local copy of the file watermarked and to wash it in real-time as required.","The former case is likely to be acceptable since it is no different to the current model.","Pay-Per-View Broadcast The nature of the watermarking process makes it possible to develop a real-time washer.","This means that protected content can be streamed to the viewer for preview.","If the viewer subsequently decides to pay for the content they can be given a key to enter into their set-top box (for example) and the content is washed locally as it arrives—there is no need to change the data being transmitted.","In order to facilitate real-time washing it may be necessary to download all the encrypted watermark information to the set-top box first.","Alternatively, the content compression process can be modified to incorporate padding in the picture frames sufficient to compensate for the additional watermark data.","With a given template design calculations can be made to determine the worst case increase that may need to be compensated.","The invention may be implemented by hardware or software or a combination of both.","The invention provides a computer program which when run on a computer implements the methods described above.","The invention also provides a store storing such a program."]],"string":"[\n [\n \"Modifying bitstreams\",\n \"The bitstream includes digital codes representing an information signal.\",\n \"At least one digital code is selected.\",\n \"The code occupies a part of the bitstream which is to contain at least one watermark code which represents a watermark perceptible in the information signal.\",\n \"The selected digital code(s) are removed from the said part of the bitstream.\",\n \"The watermark code(s) are put in the said part of the bitstream in place of the selected code(s).\",\n \"The number of bits of the selected code(s) removed from the said part of the bitstream is greater than or equal to the number of bits of the said watermark code(s) put in the said part.\",\n \"The removed selected code(s) are appended to an end of the bitstream and/or placed in watermark user data fields created in the bitstream.\"\n ],\n [\n \"1.A method of modifying a bitstream, the bitstream including digital codes representing information, the method comprising selecting at least one digital code which occupies a part of the bitstream which is to contain at least one modification code which represents a modification of the information, removing the selected digital code(s) from the said part of the bitstream, and putting the modified code(s) in the said part of the bitstream in place of the selected code(s), wherein the number of bits of the selected code(s) removed from the bitstream is greater than or equal to the number of bits of the said modification code(s) put into the bitstream; the method further comprising one or both of (1) storing the removed selected code(s) in association with the bitstream and (2) placing the removed selected codes in one or more removed code data fields in the bitstream.\",\n \"2.A method according to claim 1, comprising redistributing the selected removed codes to a removed code data field and storing the removed selected codes in association with the bitstream in a preferred order, which is: 1) within the removed code data field; and 2) storing the removed selected codes in association with the bitstream.\",\n \"3.A method according to claim 1, comprising the steps of both storing some of the removed selected codes in association with the bitstream and placing others of the removed selected codes in the said one or more removed code data fields.\",\n \"4.A method according to claim 1, wherein the step of (1) storing the removed selected code(s) in association with the bitstream comprises appending the removed selected code(s) to an end of the bitstream.\",\n \"5.A method according to claim 1, wherein the step of (1) storing the removed selected code(s) in association with the bitstream comprises storing the removed selected codes in a file.\",\n \"6.A method according to claim 5, wherein the file is appended to an end of the bitstream.\",\n \"7.A method according to claim 6, wherein the file is appended to the start end of the bitstream.\",\n \"8.A method according to claim 5, wherein the file is separate from, and associated with, the bitstream.\",\n \"9.A method according to claim 1, comprising the step of appending all the removed selected code(s) to an end of the bitstream.\",\n \"10.A method according to claim 1, comprising the step of placing all the removed selected code(s) in the said one or more removed code data fields.\",\n \"11.A method according to claim 10, comprising the step of compressing an information signal to produce the said bitstream, the amount of compression being based on a prediction of the amount of data in the modified bitstream to allow all the removed selected codes to be placed in removed code data fields within the bitstream.\",\n \"12.A method according to claim 1, comprising the step of adding padding bits to the modified bitstream so that the bitrate of the modified bitstream is unchanged relative to the original bitstream.\",\n \"13.A method according to claim 1, comprising the step of using a template to define the part or parts of the bitstream which is/are to contain modification code(s).\",\n \"14.A method according to claim 1 wherein the said digital codes representing the information are codes representing transform coefficients of the information.\",\n \"15.A method according to claim 14, wherein the coefficients are DCT coefficients.\",\n \"16.A method according to claim 15, wherein the DCT coefficients include DC and AC coefficients, all the codes representing the DC and AC coefficients are removed from the said part of the bitstream and the modification codes placed in the said part of the bitstream are codes representing DC coefficients.\",\n \"17.A method according to claim 16, wherein the codes removed from the same part of the bitstream are grouped in a group identified by a group code.\",\n \"18.A method according to claim 17, wherein the said part of the bitstream is at least one DCT block.\",\n \"19.A method according to claim 16, wherein the bitstream is modifiable by: at least one of 1) the technique of removing codes representing AC coefficients and 2) the technique of removing codes representing DC coefficients: replacing the DC codes by modified DC codes and storing the removed selected code(s) (a) in association with the bitstream and/or (b) placing them in one or more removed code data fields in the bitstream: the method further comprising the step of assessing the effects of the said techniques on the number of bits in the bitstream and selecting, in dependence on the assessment, one or more of the said techniques for performing the modification of the bitstream.\",\n \"20.A method according to claim 15, wherein the bitstream is a video bitstream which comprises at least intra encoded pictures and the method is performed on the intra-encoded pictures.\",\n \"21.A method according to claim 20, wherein the bitstream comprises inter-encoded pictures and the method is performed on the inter-encoded pictures.\",\n \"22.A method according to claim 21, wherein the inter-encoded pictures include DC codes representing DC coefficients, and comprising the steps of removing selected ones of the said DC codes, storing them in association with the bitstream and/or placing them in one or more removed code data fields in the bitstream and replacing the removed codes with modification codes of predetermined value.\",\n \"23.A method according to claim 22, wherein the said DC codes representing DC coefficients in inter-encoded pictures represent quantised change values and the said predetermined value represents the minimum quantum of change greater than zero.\",\n \"24.A method according to claim 23, wherein the said predetermined value is plus or minus one.\",\n \"25.A method according to claim 21, wherein the inter-encoded pictures include codes representing motion vectors and further comprising the steps of removing the motion vector codes associated with parts of the bitstream which are to contain modification codes, replacing the original motion vector codes with motion vector modification codes representing zero motion vectors, and appending the removed motion vector codes to the end of the bitstream and/or placing the removed codes in removed code data fields in the bitstream.\",\n \"26.A method according to claim 25, comprising the further steps of removing the motion vector codes associated with parts of the inter-encoded pictures which, in the picture, are adjacent the said parts of the bitstream which are to contain modification codes, replacing the original motion vector codes of the said associated parts with motion vector modification codes representing zero motion vectors, and appending the removed motion vector codes to the end of the bitstream and/or placing the removed codes in removed code data fields in the bitstream.\",\n \"27.A method according to claim 25, comprising providing a first template defining the blocks of intra-encoded pictures to be modified and a second template defining the blocks corresponding to the blocks of the first template of the inter-encoded pictures for which the associated motion vectors are to be set to zero.\",\n \"28.A method according to claim 27, wherein the second template defines macroblocks which include the said blocks for which the associated motion vectors are to be set to zero.\",\n \"29.A method according to claim 28, wherein the second template defines further macroblocks, for which the associated motion vectors are to be set to zero, bordering the said first-mentioned macroblocks.\",\n \"30.A method according to claim 20, wherein the bitstream comprises DCT blocks which may be encoded as field type or frame type with type codes indicating the type of encoding, and comprising the step of removing selected ones of the said type codes indicating field type and replacing the removed type codes with modification type codes indicating frame type and storing the removed type codes in association with the bitstream and/or placing them in one or more removed code data fields in the bitstream.\",\n \"31.A method according to claim 15, wherein the bitstream is an MPEG2 bitstream.\",\n \"32.A method according to claim 1, comprising the further step of providing data indicating portions of the bit stream which contain modification codes.\",\n \"33.A method according to claim 31, wherein the bitstream includes a supplementary data field, and comprising the step of providing data in the bitstream which distinguishes the removed code data field(s) from the said supplementary data field.\",\n \"34.A method according to claim 1, comprising the step of encrypting the removed selected codes before storing them in association with the bitstream and/or placing them in the said removed code data field(s).\",\n \"35.A method of restoring an original bitstream, the method comprising receiving a modified bitstream which has modification codes in selected parts of the bitstream from which original codes have been removed and stored, the method comprising the further steps of: parsing the bitstream; taking the original codes from store; and replacing the modification codes with the original codes.\",\n \"36.A method of restoring an original bitstream, the method comprising receiving a modified bitstream which has modification codes in selected parts of the bitstream from which original codes have been removed and placed in modification user data portions of the bitstream, the method comprising the further steps of: parsing the bitstream; storing the original codes; and replacing the modification codes with the stored original codes.\",\n \"37.A method of restoring an original bitstream, the method comprising receiving a modified bitstream which has modification codes in selected parts of the bitstream from which original codes have been removed and placed in modification user data portions of the bitstream and appended to an end of the bitstream, the method comprising the further steps of: parsing the bitstream; storing the original codes; and replacing the modification codes with the stored original codes.\",\n \"38.A method according to claim 36, wherein the original codes are encrypted and comprising the further step of decrypting them before replacing the modification codes with the decrypted original codes.\",\n \"39.A method according to claim 35, comprising the step of using a template which defines the part or parts of the bitstream which contain the modification code(s) to select those codes for replacement.\",\n \"40.A method according to claim 1, wherein the modification creates a perceptible effect in the information represented by the information signal.\",\n \"41.Apparatus arranged to carry out the method of claim 1.42.A computer program arranged to carry out the method of claim 1, when run on a suitable processor.\",\n \"43.A transmission medium storing a computer program according to claim 42.44.A medium according to claim 43 which is a storage medium.\",\n \"45.A medium according to claim 43 which is a communications channel.\",\n \"46.A modified bitstream which has modification codes in selected parts of the bitstream from which original codes have been removed and appended to an end of the bitstream.\",\n \"47.A modified bitstream which has modification codes in selected parts of the bitstream from which original codes have been removed and placed in modification user data portions within the bitstream.\",\n \"48.A modified bitstream which has modification codes in selected parts of the bitstream from which original codes have been removed and some placed in modification user data portions within the bitstream and others appended to an end of the bitstream.\",\n \"49.Apparatus for modifying a bitstream, the bitstream including digital codes representing information, the apparatus comprising a selecting stage operable to select at least one digital code which occupies a part of the bitstream which is to contain at least one modification code which represents a modification perceptible in the information, a removal stage operable to remove the selected digital code(s) from the said part of the bitstream, a modification code placing stage operable to put the modification code(s) in the said part of the bitstream in place of the selected code(s), wherein the number of bits of the selected code(s) removed from the bitstream is greater than or equal to the number of bits of the said modification code(s) put into the bitstream, and a removed code placing stage operable to (1) store the removed selected code(s) in association with the bitstream and/or (2) place the removed selected codes in one or more removed code data fields in the bitstream.\",\n \"50.Apparatus according to claim 49, wherein the removed code placing stage is operable to redistribute the selected removed codes to a removed code data field and store the removed selected codes in association with the bitstream in a preferred order, which is: 1) within the removed code data field; and 2) store the removed selected codes in association with the bitstream.\",\n \"51.Apparatus according to claim 49, wherein the removed code placing stage is operable to both store some of the removed selected codes in association with the bitstream and place others of the removed selected codes in the said one or more removed code data fields.\",\n \"52.Apparatus according to claim 49, wherein the removed code placing stage is operable to (1) store the removed selected code(s) in association with the bitstream by appending the removed selected code(s) to an end of the bitstream.\",\n \"53.Apparatus according to claim 49, wherein the removed code placing stage is operable to (1) store the removed selected code(s) in association with the bitstream by storing the removed selected codes in a file.\",\n \"54.Apparatus according to claim 53, wherein the removed code placing stage is operable to append the file to an end of the bitstream.\",\n \"55.Apparatus according to claim 53, wherein the removed code placing is operable to store the file separate from, but associated with, the bitstream.\",\n \"56.Apparatus according to claim 49, wherein the removed code placing stage is operable to append all the removed selected code(s) to an end of the bitstream.\",\n \"57.Apparatus according to claim 49, wherein the removed code placing stage is operable to place all the removed selected code(s) in the said one or more removed code data fields.\",\n \"58.Apparatus according to claim 49, wherein the said digital codes represent DCT coefficients of the said information.\",\n \"59.Apparatus according to claim 58, wherein the DCT coefficients include DC and AC coefficients, the removal stage is operable to remove all the codes representing the DC and AC coefficients from the said part of the bitstream and the modification code placing stage is operable to place the modification codes representing DC coefficients in the said part of the bitstream.\",\n \"60.Apparatus according to claim 58, wherein the removal stage is operable to carry out at least one of 1) a technique to remove codes representing AC coefficients and 2) a technique to remove codes representing DC coefficients: the modification code placing stage is operable to replace the removed DC codes by modified DC codes, and the removed code placing stage is operable to store the removed selected code(s) in association with the bitstream and/or place them in one or more removed code data fields in the bitstream, the apparatus further comprising a stage operable to assess the effects of the said techniques on the number of bits in the bitstream and select one or more of the said techniques in dependence on the assessment.\",\n \"61.Apparatus according to claim 60 wherein the removed code placing stage is operable to append the removed selected code(s) to an end of the bitstream and/or place them in one or more removed code data fields in the bitstream.\",\n \"62.Apparatus according to claim 59, wherein the bitstream comprises at least intra encoded pictures and the apparatus is arranged to modify the intra-encoded pictures.\",\n \"63.Apparatus according to claim 62, wherein the bitstream comprises inter-encoded pictures and the apparatus is arranged to additionally modify the inter-encoded pictures.\",\n \"64.Apparatus according to claim 63, wherein the inter-encoded pictures include codes representing DC coefficients, the apparatus comprising a stage operable to remove selected ones of the said codes, and a placing stage operable to store them in association with the bitstream and/or place them in one or more removed code data fields in the bitstream and a replacing stage operable to replace the removed codes with modification codes of predetermined value.\",\n \"65.Apparatus according to claim 64, wherein the said codes representing DC coefficients in inter-encoded pictures represent quantised change values and the said predetermined value represents the minimum quantum of change greater than zero.\",\n \"66.Apparatus according to claim 65, wherein the said predetermined value is plus or minus one.\",\n \"67.Apparatus according to claim 63, wherein the inter-encoded pictures include codes representing motion vectors and further comprising a motion vector code removing stage operable to remove the motion vector codes associated with parts of the bitstream which are to contain modification codes, replace the original motion vector codes with motion vector modification codes representing zero motion vectors, and store the removed motion vector codes in association with the bitstream and/or placing the removed codes in removed code data fields in the bitstream.\",\n \"68.Apparatus according to claim 67, wherein the motion vector code removing stage is further operable to remove the motion vector codes associated with parts of the inter-encoded pictures which, in the picture, are adjacent the said parts of the bitstream which are to contain modification codes, replace the original motion vector codes of the said associated parts with motion vector modification codes representing zero motion vectors, and store the removed motion vector codes in association with the bitstream and/or place the removed codes in removed code data fields in the bitstream.\",\n \"69.Apparatus according to claim 58, wherein the bitstream comprises DCT blocks which may be encoded as field type or frame type with type codes indicating the type of encoding, and comprising a removal stage operable to remove selected ones of the said type codes indicating field type and a replacing stage operable to replace the removed type codes with modification type codes indicating frame type and a removed code placing stage operable to store the removed type codes in association with the bitstream and/or place them in one or more removed code data fields in the bitstream.\",\n \"70.Apparatus for restoring an original bitstream, the apparatus comprising a receiving stage operable to receive a modified bitstream which has modification codes in selected parts of the bitstream from which original codes have been removed and stored, a parsing stage operable to parse the bitstream; a store operable to store the original codes; and a replacing stage operable to replace the modification codes with the original codes.\",\n \"71.Apparatus for restoring an original bitstream, the apparatus comprising a receiving stage operable to a receive a modified bitstream which has modification codes in selected parts of the bitstream from which original codes have been removed and placed in modification user data portions of the bitstream: a parsing stage operable to parse the bitstream; a store for storing the original codes; and a replacing stage operable to replace the modification codes with the stored original codes.\",\n \"72.Apparatus for restoring an original bitstream, comprising a receiving stage operable to receive a modified bitstream which has modification codes in selected parts of the bitstream from which original codes have been removed and placed in modification user data portions of the bitstream and appended to an end of the bitstream, a parsing stage operable to parse the bitstream; a store for storing the original codes; and a replacing stage operable to replace the modification codes with the stored original codes.\",\n \"73.Apparatus according to claim 71, for use with a bitstream wherein the original codes are encrypted and comprising a decrypting stage operable to decrypt the original codes before replacing the modification codes with the decrypted original codes.\",\n \"74.A data carrier storing a template defining the part or parts of a bitstream which is/are to contain modification code(s) and at least one decryption key.\",\n \"75.The invention of claim 74 wherein the information signal includes one or more of: video data; audio data; audio/visual data; and other data.\",\n \"76.A method of visibly watermarking a video sequence having intra-encoded pictures and inter-encoded pictures, the pictures comprising data representing blocks of transform coefficients of the video, motion vectors being associated with at least some of the blocks of the inter-encoded pictures, the method comprising providing at least one template defining the blocks to be modified to visibly watermark the video, modifying at least some transform coefficients in the blocks defined by the template(s) in at least the intra-encoded pictures to visibly watermark the video, and setting to zero the motion vectors associated with blocks of the inter-encoded pictures defined by the template(s) and corresponding to the modified blocks of the intra-encoded pictures.\",\n \"77.A method according to claim 76, comprising providing a first template defining the blocks of intra-encoded pictures to be modified and a second template defining the blocks corresponding to the blocks of the first template of the inter-encoded pictures for which the associated motion vectors are to be set to zero.\",\n \"78.A method according to claim 77, wherein the second template defines macroblocks which include the said blocks for which the associated motion vectors are to be set to zero.\",\n \"79.A method according to claim 78, wherein the second template defines further macroblocks, for which the associated motion vectors are to be set to zero, bordering the said first-mentioned macroblocks.\",\n \"80.A method according to claim 76, wherein the video sequence is represented by a bitstream including digital codes representing the blocks and motion vectors, the method comprising selecting the digital codes which represent the blocks to be modified and the motion vectors to be set to zero, removing the selected digital code(s) from the bitstream, and putting modified codes in the bitstream in place of the selected codes, the method further comprising one or both of (1) appending the removed codes to an end of the bitstream and (2) placing the removed codes in one or more removed code data fields in the bitstream.\",\n \"81.A method according to claim 80, further comprising encrypting the removed codes.\",\n \"82.Apparatus arranged to carry out the method of claim 76.83.A computer program which when run on a suitable data processor implements the method of claim 76.84.A data carrier on which is stored a program according to claim 83.85.Apparatus for visibly watermarking a video sequence having intra-encoded pictures and inter-encoded pictures, the pictures comprising data representing blocks of transform coefficients of the video, motion vectors being associated with at least some of the blocks of the inter-encoded pictures, the apparatus comprising a storage means for storing at least one template defining the blocks to be modified to visibly watermark the video, a modifier arranged to modify at least some transform coefficients in the blocks defined by the template(s) in at least the intra-encoded pictures to visibly watermark the video, and a means arranged to set to zero the motion vectors associated with blocks of the inter-encoded pictures defined by the template(s) and corresponding to the modified blocks of the intra-encoded pictures.\",\n \"86.Apparatus according to claim 85, wherein the storage means stores a first template defining the blocks of intra-encoded pictures to be modified and a second template defining the blocks corresponding to the blocks of the first template of the inter-encoded pictures for which the associated motion vectors are to be set to zero.\",\n \"87.Apparatus according to claim 86, wherein the second template defines macroblocks which include the said blocks for which the associated motion vectors are to be set to zero.\",\n \"88.Apparatus according to claim 87, wherein the second template defines further macroblocks, for which the associated motion vectors are to be set to zero, bordering the said first-mentioned macroblocks.\",\n \"89.Apparatus according to claim 85, wherein the video sequence is represented by a bitstream including digital codes representing the blocks and motion vectors, the apparatus comprising means for selecting the digital codes which represent the blocks to be modified and the motion vectors to be set to zero, means for removing the selected digital code(s) from the bitstream, and putting modified codes in the bitstream in place of the selected codes, and means for (1) appending the removed codes to an end of the bitstream and/or (2) placing the removed codes in one or more removed code data fields in the bitstream.\",\n \"90.A method of restoring an original bitstream visibly watermarked by the method of claim 80, the modified bitstream having modified codes representing modified blocks and motion vectors set to zero in selected parts of the bitstream from which original codes have been removed and placed in modification user data portions of the bitstream and/or appended to the end of the bitstream, the method comprising the further steps of: parsing the bitstream; storing the original codes; and replacing the modified codes with the stored original codes.\",\n \"91.A method according to claim 90, wherein the original codes are encrypted and comprising the further step of decrypting them before replacing the modification codes with the decrypted original codes.\",\n \"92.Apparatus for restoring an original bitstream, comprising: a receiving stage operable to receive a modified bitstream visibly watermarked by the method of claim 5, the modified bitstream having modified codes representing modified blocks and motion vectors set to zero in selected parts of the bitstream from which original codes have been removed and placed in modification user data portions of the bitstream and/or appended to the end of the bitstream the apparatus comprising a parsing stage operable to parse the bitstream; a store operable to store the original codes; and a replacing stage operable to replace the modified codes with the original stored codes.\",\n \"93.Apparatus according to claim 92, wherein the original codes are encrypted and comprising the further a stage for decrypting them before replacing the modified codes with the decrypted original codes.\",\n \"94.Apparatus arranged to carry out the method of claim 90.95.A computer program which when run on a suitable data processor implements the method of claim 90.96.A data carrier on which is stored a program according to claim 95.97.\",\n \"(canceled) 98.\",\n \"(canceled) 99.\",\n \"(canceled) 100.\",\n \"(canceled)\"\n ],\n [\n \"The present invention relates to a method of modifying a bitstream, apparatus for modifying a bitstream, a computer program for modifying a bitstream, a storage medium storing such a program, method of removing a modification from a bitstream, apparatus for removing a modification from a bitstream, a computer program for removing a modification from a bitstream, a storage medium storing such a program, a modified bitstream and a data carrier carrying modification removal data.\",\n \"It is known to provide a watermark in video, audio, audio/visual and/or other information signals.\",\n \"The watermark is provided to check the provenance of the information signal and/or to identify the owner or other persons associated with the information signal.\",\n \"A watermark may be perceptible or hidden.\",\n \"Preferred embodiments of the present invention use perceptible watermarks.\",\n \"It is also known to watermark an image by transforming the image from the spatial domain to a transform domain, e.g.\",\n \"to the wavelet domain and embed a watermark by changing the wavelet coefficients.\",\n \"The transform domain image with the watermark is then inverse transformed to the spatial domain.\",\n \"See for example “An image watermarking method based on the wavelet transform” by Hisashi Inoue, et al, IEEE, 0-7803-5467-2/99.U.S.\",\n \"Pat.\",\n \"No.\",\n \"5,809,139 (Girod et al) discloses the invisible watermarking of video material which is already compressed.\",\n \"Video which has been compressed by MPEG2 is entropy decoded and dequantised to obtain the DCT coefficients.\",\n \"A spread spectrum spatial domain watermark is transformed to the DCT domain and the transformed watermark added to the DCT coefficients of the transformed video.\",\n \"To maintain the bit rate of the compressed video even with the addition of the watermark, various steps are taken.\",\n \"Only non-zero coefficients are modified.\",\n \"Also the number of bits of an encoded watermarked coefficient is compared against the number of bits of the coefficient prior to watermarking (plus any extra number of spare bits saved in previous coding operations).\",\n \"A control controls selection of the output between the watermarked bitstream and the original unwatermarked bitstream.\",\n \"If an entropy encoded unwatermarked coefficient uses n0 bits and the watermarked entropy encoded coefficient uses n1 bits then a watermarked coefficient is output if n1<=n0+n3, where n3 is the number of spare bits available for use.\",\n \"The watermark can be detected in a video decoder but it is not possible to restore the watermarked video to the original video by reversing the watermarking algorithm.\",\n \"WO 99/10837 (Digimarc) discloses a method of watermarking a video image in which the image is compressed using for example MPEG2 and the DCT coefficients are modified to embed a watermark.\",\n \"The bit rate is preserved by maintaining a count that represents the amount the bit rate has been increased by the modifications to the coefficients less the amount the bit rate has been decreased by the modifications.\",\n \"If the cumulative change exceeds a positive or negative limit, then no further changes are made.\",\n \"The watermark can be detected in a video decoder but it is not possible to restore the watermarked video to the original video by reversing the watermarking algorithm.\",\n \"Both U.S. Pat.\",\n \"No.\",\n \"5,809,139 and WO 99/10837 require means which monitor the bit rate and which stops embedding of a watermark if a specified limit is reached.\",\n \"For US-A-5809139 and WO 99/10837 that results in a random distribution of the watermark in an image.\",\n \"These proposals are aimed at irreversible imperceptible watermarks which embed data representing a message in the image.\",\n \"It is known to compression encode information signals especially image signals.\",\n \"Also, if a modification is made to the information signals which adds data to the information signals, then the compression is usually less efficient.\",\n \"It is desirable to process compressed information signals, so as to produce modification of the information signals which is perceptible in the decompressed information signals, e.g.\",\n \"a visible modification in an image, with a minimal change to the number of bits representing the compressed information signals.\",\n \"Copending prior application EP 01310113.4 (U.S. Ser.\",\n \"No.\",\n \"10/006,050) is an example of a method and apparatus for modifying a compressed video bitstream and for removing the watermark therefrom.\",\n \"A compression process carried out on the bitstream uses DCT coefficients.\",\n \"The modifying process is carried out to ensure the watermarked bitstream has no more bits than, and thus the same bit rate as, the bitstream before it was watermarked.\",\n \"That is done by encrypting fixed length parts of codes representing DCT coefficients without increasing the number of bits in the fixed length codes.\",\n \"The most preferred embodiment is a compressed bitstream of I frames and the technique works well with such a bitstream to produce a visible watermark.\",\n \"The technique can be applied to other compressed bitstreams but the visibility of the watermark in the displayed image may not be as desired due to the limitation of not increasing the number of bits.\",\n \"MPEG2 is well known and is defined in ISO/IEC/13818.The invention seeks to provide such a technique which is applicable to bitstreams containing video, audio, audio/visual and/or other information signals and which produces a modification, preferably a removable and perceptible modification, without increasing the bitrate, and preferably without changing the bit rate, of the information signal.\",\n \"An embodiment of the present invention seeks to provide a modifying technique which can be applied to a compressed bitstream, (e.g.\",\n \"MPEG2) and which can produce a modification of desired perceptibility, whilst at the same time not changing the bit rate in the compressed bitstream and which modification is removable from the bitstream.\",\n \"One aspect of the invention provides a method of modifying a bitstream, the bitstream including digital codes representing information, the method comprising selecting at least one digital code which occupies a part of the bitstream which is to contain at least one modification code which represents a modification of the information, removing the selected digital code(s) from the said part of the bitstream, and putting the modified code(s) in the said part of the bitstream in place of the selected code(s), wherein the number of bits of the selected code(s) removed from the bitstream is greater than or equal to the number of bits of the said modification code(s) put into the bitstream; the method further comprising one or both of (1) storing the removed selected code(s) in association with the bitstream and (2) placing the removed selected codes in one or more removed code data fields in the bitstream.\",\n \"Preferably, the modification is such as to be perceptible in the information represented by the bitstream.\",\n \"According to a second aspect of the present invention there is provided a method of modifying a bitstream, the bitstream including digital codes representing information, the method comprising selecting at least one digital code which occupies a part of the bitstream which is to contain at least one modifying code which represents a modification perceptible in the information, removing the selected digital code(s) from the said part of the bitstream, putting the modification code(s) in the said part of the bitstream in place of the selected code(s), wherein the number of bits of the selected code(s) removed from the bitstream is greater than or equal to the number of bits of the said modification code(s) put in the said part, and appending the removed selected code(s) to an end of, but not within, the bitstream According to a third aspect of the present invention, there is provided a method of modifying a bitstream having an information signal portion including digital codes representing information, the method comprising selecting digital codes which occupy a part of the said information signal portion which is to contain at least one modification code which represents a modification perceptible in the information, removing the selected digital codes from the said part of the bitstream, putting the modification code(s) in the said part of the bitstream in place of the selected codes, creating a modification user data portion of the bitstream appended with the said part and storing at least one of the removed selected code(s) in the said modification user data portion, wherein the number of bits in the said information signal portion containing the modification codes plus the number of bits in the said modification user data portion is less than or equal to the number of bits in the said information signal portion before removal of the selected codes.\",\n \"The modification in embodiments of the invention is the provision of a perceptible modification of the information represented by bitstream rather than the hiding of data within the bitstream.\",\n \"In this way, the bitrate of the modified bitstream is maintained equal to that of the original bitstream.\",\n \"The addition of the modification codes produces additional data.\",\n \"By removing the original codes and appending them to the end of the bitstream, the amount of data in the bitstream is not increased.\",\n \"Spare space created in the bitstream by the removal of the original codes may be used as the modification user data fields for at least some of the original codes and any codes which cannot be included in those fields are appended to an end of the bitstream.\",\n \"The codes may be appended to the start end of the bitstream or to the finish end.\",\n \"The codes may be in a separate file or in a file appended to the start end or the finish end of the bitstream.\",\n \"The bitstream in preferred examples of the invention is a compressed bitstream preferably compressed according to the MPEG2 standard.\",\n \"It is particularly important in MPEG2 that the bitrate of the bitstream conforms to the standard.\",\n \"Thus the present invention makes possible that the MPEG2 bitstream even with the modification codes in it complies with the standard and any excess data resulting from the modifying process is not in the MPEG2 compliant bitstream but appended to it.\",\n \"Excess data is the data which cannot be placed in the modification user data fields in the bitstream.\",\n \"Preferred embodiments of the present invention allow all the original data to be preserved allowing removal of the modification data and restoration of the original bitstream.\",\n \"(This assumes, of course, that data is not lost due to other processing of the modified bitstream for example transmission via a noisy channel or further processing of the modified signal.)\",\n \"A fourth aspect of the invention provides an apparatus for modifying a bitstream, the bitstream including digital codes representing information, the apparatus comprising a selecting stage operable to select at least one digital code which occupies a part of the bitstream which is to contain at least one modification code which represents a modification perceptible in the information, a removal stage operable to remove the selected digital code(s) from the said part of the bitstream, a modification code placing stage operable to put the modification code(s) in the said part of the bitstream in place of the selected code(s), wherein the number of bits of the selected code(s) removed from the bitstream is greater than or equal to the number of bits of the said modification code(s) put into the bitstream, and a removed code placing stage operable to (1) store the removed selected code(s) in association with the bitstream and/or (2) place the removed selected codes in one or more removed code data fields in the bitstream.\",\n \"In an embodiment of the invention the bitstream comprises digital codes representing transform coefficients of the information signal, for example DCT coefficients, including DC and AC coefficients.\",\n \"An example of the invention uses various techniques for modifying such a bitstream.\",\n \"Those techniques include 1) removing AC coefficients and 2) removing DC coefficients.\",\n \"The removed coefficients are replaced by modification codes.\",\n \"The effects of the various techniques on the number of bits in the modified bitstream is assessed and one or more of the techniques is selected for the actual modification of the bitstream in dependence on the assessment.\",\n \"This allows the techniques to be selected to maintain the bitrate of the modified bit stream equal to that of the original bitstream.\",\n \"The original bitstream may include inter-encoded pictures including codes representing motion vectors.\",\n \"If the modification represents a visible watermark in a picture, the motion vectors distort the watermark.\",\n \"An example of the invention, comprises the steps of removing the motion vector codes associated with the said part of the bitstream which is to contain at least one modification code, replacing the original motion vector codes with motion vector modification codes representing zero motion vectors, and placing the removed motion vector codes at the end of the bitstream and/or in removed code data fields in the bitstream.\",\n \"This reduces distortion of the watermark.\",\n \"MPEG2 bitstreams may include DCT blocks encoded as field type or frame type with type codes indicating the type of encoding.\",\n \"It has been found that if a field type DCT block is modified but an adjacent field type block is not modified the watermark is distorted by stripes.\",\n \"Thus an example of the invention comprises the step of removing selected ones of the said type codes indicating field type and replacing the removed type codes with modification type codes indicating frame type and placing the removed type codes in one or both of (1) the end of the bitstream and (2) in one or more removed code data fields in the bitstream.\",\n \"Other aspects of the invention are specified in the claims to which attention is invited.\",\n \"For a better understanding of the present invention, and to show how the same may be carried into effect, reference will now be made, by way of example, to the accompanying drawings in which: FIG.\",\n \"1 is a schematic diagram of an image frame; FIG.\",\n \"2 is a schematic diagram of blocks of 8×8 pixels in the image frame; FIG.\",\n \"3 is a schematic diagram of a DCT block; FIG.\",\n \"4 is a schematic diagram of macroblocks; FIG.\",\n \"5 is a schematic diagram of rows of macroblocks and slices; FIG.\",\n \"6 is a schematic diagram of a Group of Pictures (GOP); FIG.\",\n \"7 is a schematic diagram of one frame of picture data; FIG.\",\n \"8 is a schematic diagram of a bitstream of one macroblock; FIG.\",\n \"9 is a schematic diagram of a template defining a watermark; FIG.\",\n \"10 is a flow diagram illustrating an overview of an illustrative watermarking method applied to intra-frames according to the present invention; FIGS.\",\n \"11 to 13 are schematic diagrams of illustrative data formats used in examples of the present invention; FIG.\",\n \"14 is schematic diagram showing user data distributed amongst compressed picture frames; FIG.\",\n \"15 is a flow diagram of a watermark encryption/decryption process; FIG.\",\n \"16 is a flow diagram illustrating an overview of an illustrative watermark removal method according to the present invention; FIG.\",\n \"17 is an overview of an illustrative watermarking system according to the present invention; FIG.\",\n \"18 is a schematic block diagram of a watermark removal apparatus; FIG.\",\n \"19 is a schematic diagram of three buffers used in the watermarking process and in the watermark removal process; FIG.\",\n \"20 is a flow diagram illustrating an overview of an illustrative watermarking method applied to non-intra-frames according to the present invention; FIG.\",\n \"21A illustrates watermarked macroblocks and FIG.\",\n \"21B is a flow diagram of a DCT type change process; FIG.\",\n \"22 is a flow diagram of a technique selection process; FIG.\",\n \"23 illustrates templates used in association with Motion Vectors; FIG.\",\n \"24 is a flow diagram illustrating an overview of an illustrative method applied to motion vector codes in an embodiment of the present invention; FIG.\",\n \"25 illustrates the coding of motion vectors; FIG.\",\n \"26 illustrates watermarking a video sequence of frame pictures; FIG.\",\n \"27 illustrates a) watermarking a video sequence of field pictures, and b) watermarking a video sequence of field and frame pictures; and FIG.\",\n \"28 illustrates watermarked macroblocks for interlaced fields and for a frame; FIGS.\",\n \"29 to 31 illustrates modifications of the invention.\",\n \"In the following illustrative description, an MPEG2 compressed bitstream is modified.\",\n \"The modification is referred to as a watermark.\",\n \"Applying the watermark to the bitstream is referred to as embedding.\",\n \"Restoring the original bitstream is referred to as washing.\",\n \"FIRST EXAMPLE A first example of the present invention will now be described.\",\n \"For ease of explanation, the example describes the placing of watermark data into I frames of an MPEG2 video bitstream which is a single stream with no audio; i.e.\",\n \"an MPEG2 video sequence file.\",\n \"Alternatively, the bitstream may be a packetised bitstream.\",\n \"However the invention is not limited to I frames nor is it limited to video nor is it limited to MPEG2.Also for ease of explanation, the following discussion refers only to luminance information.\",\n \"However the invention is not limited to luminance information.\",\n \"Firstly, MPEG2 will be reviewed.\",\n \"Referring to FIG.\",\n \"1, an image frame 2 is shown.\",\n \"The image is represented by digital pixels.\",\n \"As shown in FIG.\",\n \"2 blocks 4 of 8×8 pixels are selected and as shown in FIG.\",\n \"3 those blocks are subject to a Discrete Cosine Transform (DCT) to form DCT blocks containing 8×8 DCT coefficients.\",\n \"The coefficients include a DC coefficient 6 and AC coefficients 7.To form a serial bitstream the coefficients are scanned in a zig-zag scan pattern 8 as illustrated in FIG.\",\n \"3.\",\n \"(Other patterns are known).\",\n \"Groups of four adjacent DCT blocks are grouped as macroblocks 10 as shown in FIG.\",\n \"4.As shown in FIG.\",\n \"5, the image frame is organised as rows R1, R2, R3 etc.\",\n \"of macroblocks.\",\n \"The macroblocks are organised into slices.\",\n \"A slice is a horizontal collection of macroblocks.\",\n \"A new slice starts at the beginning of each row of macroblocks.\",\n \"A slice may contain any number of macroblocks.\",\n \"A row of macroblocks may contain any number of slices.\",\n \"Referring to FIG.\",\n \"6, MPEG2 has the following types of frame.\",\n \"An I frame is an intra encoded frame; that is a frame in which all the image data is contained in that frame without reference to any other frame.\",\n \"A P frame is a frame that is coded using motion compensated prediction from a previous I frame or P frame.\",\n \"A B frame is a frame coded using motion compensated prediction from a previous and/or future I frame or P frame.\",\n \"For ease of explanation the following description of the first example refers only to I frames.\",\n \"Frames are organised into Groups of Pictures called GOPs.\",\n \"FIG.\",\n \"6 shows part of a typical GOP in which a repeated sequence of I, B, B and P frames form the GOP.\",\n \"Other sequences are possible.\",\n \"The number of frames in a typical GOP is 12 or 15, but other numbers of frames are possible.\",\n \"FIG.\",\n \"7 and FIG.\",\n \"11 illustrates the data format of what is referred to herein as picture data.\",\n \"Picture data herein includes all data between picture start codes and includes headers (HDR), userdata (UD) and the image data 16 representing one frame of the image.\",\n \"MPEG2 allows user data (UD) to be placed in the picture data ahead of the image data 16.There may be no user data fields UD or one or more user data fields.\",\n \"The DCT blocks and macroblocks are formatted as shown in FIG.\",\n \"8 in the image data 16.Each block includes a DC coefficient followed by AC coefficients.\",\n \"Blocks are separated by End of Block codes EOB.\",\n \"In I frames the DC coefficients are differentially encoded.\",\n \"The differential values are referred to herein as DC differentials.\",\n \"The AC coefficients are run length encoded and represented by Variable Length Codes (VLCs) and are referred to herein as AC VLCs.\",\n \"The MPEG2 encoding process must produce a bitstream which is decodable by all decoders which comply with the MPEG2 standard.\",\n \"Decoders include a buffer which temporarily stores the bitstream as it is decoded in the buffer.\",\n \"The MPEG2 standard defines the size of the buffer.\",\n \"Generally, the buffer should neither underflow nor overflow otherwise data is lost.\",\n \"(Underflow may be allowed under certain conditions) The encoding process is carried out such that the bit rate is constrained to prevent underflow and overflow.\",\n \"Modifying an encoded bitstream, in this example of the invention for the purpose of applying a visible watermark to it, risks changing the bit rate and thus risks underflow or overflow.\",\n \"The example of the invention described hereinafter seeks to apply a visible watermark to the bitstream without changing the bit rate of the picture data and thus without causing the buffer to underflow or overflow and to allow the watermark to be removed and the original image restored.\",\n \"Referring to FIG.\",\n \"9, a template 18 defines the watermark.\",\n \"In the example of FIG.\",\n \"9 the watermark is in the form of an S as in Sony but the watermark could have any other form.\",\n \"The template defines the watermark to the resolution of a DCT block Templates are further discussed below in the section headed “Templates”.\",\n \"Referring to FIG.\",\n \"10, in overview, the first example of the invention may operate as follows.\",\n \"These operations are performed only on I frames in this example but the invention is not limited to performing these operations on I frames.\",\n \"In step S2, the macroblocks which fall within the watermark S are selected and all the DC differential and AC VLCs of those macroblocks are removed and in step S4 stored.\",\n \"In step S6, new DC differentials which represent the watermark are put into the blocks of the template macroblocks in place of the ones which were removed.\",\n \"(The AC VLCs are not replaced in those blocks).\",\n \"The DC coefficients in I frames are differentially encoded and so the new DC differentials need to be calculated in dependence on the DC differentials of the macroblocks spatially preceding and following the macroblocks at the edges of the template as will be described in more detail below.\",\n \"In step S8 the number of bits Nr removed from an I frame by removing the original DC differentials and AC VLCs is determined and the number of bits Na added to the I frame by the new DC differentials is determined.\",\n \"The difference Nr-Na indicates the amount of spare data space in the picture data.\",\n \"If that space exceeds a threshold amount (5 bytes for MPEG2) then referring to FIG.\",\n \"11B a visible watermark user data space 20 is created in the picture data.\",\n \"At least some of the stored DC differentials and AC VLCs are placed in that data space 20.To ensure that the bit rate of the picture data is not changed zero padding 22 may be added to the picture data.\",\n \"If insufficient spare space is available to create the data space 20 then zero padding 22 is added.\",\n \"The coefficients placed in the data space 20 are encrypted before they are placed in the space 20 to ensure that only authorised persons can reconstruct the original image.\",\n \"That process is repeated (S10) for each I frame in a video sequence.\",\n \"Referring to FIG.\",\n \"14, a portion of a video sequence is shown.\",\n \"FIG.\",\n \"14 shows only I frames for ease of explanation.\",\n \"The picture data frames are shown in simplified form for ease of explanation.\",\n \"If the sequence comprises P and/or B frames, the I frames would be separated by such frames.\",\n \"The first I frame P1 of the sequence includes watermark user data space 20 which contains some of the removed coefficients of frame P1.Frame P2 has a data space 20 and the other coefficients removed from frame P1 are in that data space.\",\n \"In the example of FIG.\",\n \"14, the user data space 20 of frame P2 has capacity to contain some of the removed coefficients of the frame P2.The others of the removed coefficients of frame P2 are in the user data space 20 of frame P3.Frame P4 is shown as having no spare capacity for a user data space 20 and some of the coefficients removed from frame P3 are in the user data space 20 of frame P5.Thus as illustrated by FIG.\",\n \"14, the coefficients removed in serial order from the image data space of the frames of a video sequence are taken from store, encrypted and put, in the same serial order, into watermark user data spaces 20 created in the frames of picture data but without increasing the bit rate of the picture data.\",\n \"The end of the video sequence is indicated by an end of sequence code 26.The user data spaces created in the video sequence may not provide sufficient spare capacity to contain all the removed coefficients.\",\n \"Thus as shown in step S12 of FIG.\",\n \"10, the remaining coefficients are taken from the store, encrypted and appended to the end of the video sequence after the end of sequence code 26 as shown at 28 in FIG.\",\n \"14.The data appended to the end of the MPEG2 bitstream is part of the MPEG2 bitstream but because it is not prefaced by a start code, the appended data is ignored by an MPEG2 decoder.\",\n \"Although the process has so far been described only with reference to I frames, the video sequence typically also includes P and B frames.\",\n \"Some P and B frames may include zero padding.\",\n \"If so the zero padding may be removed and used for a watermark user data block and the removed original coefficients may be placed in the user data block.\",\n \"The zero padding is reinstated in the process of restoring the original bitstream.\",\n \"That overview omits some details of operation which will be discussed in the following discussion.\",\n \"End of Block and Other Codes As best shown in FIG.\",\n \"13, the original data 30 (DC differentials and AC VLCs) removed from the image data 16 includes the original (or copies thereof) end of block codes EOB.\",\n \"The data 32 (i.e.\",\n \"the new DC differentials) added to the image data 16 includes new (or the original) end of block codes EOB.\",\n \"In addition the newly created watermark user data spaces 20 include userdata start codes 34 which increase the number of bits in the picture data frames.\",\n \"When determining the spare capacity available in the picture data frames once the original AC and DC differentials have been removed only the DC differentials and AC variable length codes are included since the EOB codes remain.\",\n \"Flagging Watermark Userdata Blocks Referring to FIG.\",\n \"12, it is necessary to be able to distinguish a watermark userdata block 20 inserted by the watermarking process from any user data block originally present.\",\n \"For this purpose, the following technique is used.\",\n \"Regardless of whether a picture frame contains an original userdata block 36, if there is space within the picture frame to include a watermark userdata block 20 the watermark userdata block 20 is the last userdata block in the picture frame.\",\n \"See FIG.\",\n \"12B.\",\n \"The last bit 38 of the last byte of the watermark userdata is set to ‘1’ (i.e.\",\n \"send a whole number of bytes up to the last byte in the userdata then only send 7-bits of watermark data followed by a single ‘1’ bit).\",\n \"As shown in FIG.\",\n \"12C if a picture contains one or more original userdata blocks 36 and there is no space within the picture to include a watermark userdata block then the last bit 40 of the last byte of the last block of userdata is stored in the watermark data and replaced by a ‘0’.\",\n \"If a picture contains no original userdata blocks and there is no space within the picture to include a watermark userdata block then no operation is required.\",\n \"Changing the last bit as described above is only an example.\",\n \"The first bit could be changed instead.\",\n \"Encryption of Watermark Data, FIG.\",\n \"15 The watermark data as it appears in the watermarked MPEG-2 file must not contain MPEG start codes since they will terminate userdata areas prematurely or cause other undesirable decoder behaviour.\",\n \"The nature of the codes used to represent DC differentials and AC VLCs is such that start codes cannot be accidentally generated within the displayed image data.\",\n \"It should not therefore be possible to generate them in the watermark data either providing care is exercised when generating flag information.\",\n \"However, the encryption of the watermark data may result in start codes being produced.\",\n \"It is therefore necessary to check the encrypted data for start codes and where they occur revert to the unencrypted watermark data (this does not affect security since it is only possible to determine which bytes have been left unencrypted when decrypting with the correct key).\",\n \"Where it has been necessary to revert to the unencrypted watermark data, a check must also be performed to ensure that this did not result in the generation of another start code across the encrypted/unencrypted data boundary.\",\n \"Any suitable encryption algorithm may be used.\",\n \"Currently the AES algorithm is preferred.\",\n \"FIG.\",\n \"15 shows an illustrative encryption process using an Exclusive OR (XOR) operation.\",\n \"Because the XOR operation is used, the decryption process is identical to the encryption process.\",\n \"In FIG.\",\n \"15: “idx” is an index count indexing the watermark data bytes to be encrypted (or decrypted); “zero_count” is a count of the number of consecutive zeros in the bitstream; “ubyte” is a byte of unencrypted watermark data; “ebyte” is a byte of encrypted watermark data; “r” is a random number generated by the encryption algorithm and is different for each byte; “zero_count++” means increment zero_count; and “idx++” means increment idx.\",\n \"In overview, the encryption process is monitored to ensure that an MPEG start code is not generated by chance.\",\n \"An MPEG start code is for example 3 hexadecimal bytes 00 00 01.Such a code is not generated in the unencrypted watermark data but it is possible it would occur by chance in the encrypted data.\",\n \"If such a code is generated in the encrypted data the unencrypted data is used instead.\",\n \"The process of FIG.\",\n \"15 checks for a sequence of zeros (step S57) or a sequence of zeros followed by a one (step S58) in which case it outputs (S60) the unencrypted data ubyte, otherwise it outputs (S59) the encrypted data ebyte.\",\n \"Referring to FIG.\",\n \"15, the encryption (or decryption) process starts at step S50 and proceeds to step S51 in which idx and zero_count are set to zero.\",\n \"In step S52, idx is compared to the number num_bytes of bytes of watermark data to be encrypted (decrypted).\",\n \"If all the bytes have been processed, the process ends at step S53, otherwise it proceeds to step S54 which gets the (next) watermark data byte ubyte to be processed and step S55 which gets a random number r for encrypting the data byte.\",\n \"In step S56 the data byte is encrypted by performing ebyte=ubyteXORr.\",\n \"Steps S57 and S58 check whether there is likely to be an MPEG start code in the resulting data stream.\",\n \"Step S57 checks whether ubyte or ebyte is a sequence of zeros in which case step S59 outputs ubyte.\",\n \"Step S58 checks whether there is a sequence of zeros (zero-count >1) followed by a one (ebyte=1).\",\n \"If yes, ubyte is output S60, otherwise ebyte is output S59.At step S61, if ubyte is zero, then the zero_count is incremented, otherwise zero count is set to zero in step S62 and finally at step S64 idx is incremented and the process returns to step S52.Differential DC Recoding The DC coefficients for intra blocks are coded differentially throughout a slice.\",\n \"The original differential value is stored and a new differential value calculated and used to replace it as described above.\",\n \"This allows the change to be made to provide a desired, e.g.\",\n \"fixed, luminance level and gives good definition of the template design.\",\n \"Referring to FIG.\",\n \"4, assume an I frame is being modified, and assume the central macroblock 10 is the macroblock in the template to contain watermark data and the left and right hand macroblocks are outside the template.\",\n \"The DC differential 60 of the left hand block is outside the template and is not affected by the watermark.\",\n \"DC differentials 61 to 64 are in the template.\",\n \"DC differentials 61 to 64 are removed from the macroblock and stored.\",\n \"DC differentials 61 to 64 are replaced by new differential values representing the watermark.\",\n \"Assume, for ease of explanation that the watermark is an area of white level.\",\n \"DC differential 61 is replaced by a new differential value representing the desired white level and DC differential 62 to 64 which all represent the same white level as DC differential 61 are thus set to zero.\",\n \"DC differential 65 is recalculated.\",\n \"Exiting the template involves replacing the original differential value (i.e.\",\n \"of block 65) with a new one.\",\n \"There is no need to store the original differential value (of block 65) as the process can be reversed in the washer because it can be calculated from the difference between: a) the value of block 65 calculated from the watermarked differential values converted to non-differential values; and b) the original (washed) non-differential value of block 64.FIG.\",\n \"4 assumes a template coincides with a macroblock.\",\n \"That is advantageous in that it minimises the number of entry and exit points and maximises the number of zero DC differentials.\",\n \"However, a template may not coincide with a macroblock.\",\n \"P and B Frames Intra macroblocks in P and B frames can be modified as described above.\",\n \"Non-intra coded macroblocks can have the AC VLCs removed as described above but non-intra encoded macroblocks are predictively encoded and so the changes are less perceptible.\",\n \"Motion vector watermarking can only be performed on P and B frames.\",\n \"Chrominance The foregoing description assumes watermark data is provided only in luminance.\",\n \"However macroblocks may also include DCT blocks of chrominance information.\",\n \"Watermark data may be included in the chrominance DCT blocks in the same way as described for luminance.\",\n \"Also the watermark data may be included only in chrominance DCT blocks and not in luminance blocks but that is not currently preferred.\",\n \"Overview of Restoring Original Image Data (FIG.\",\n \"16) Referring to FIG.\",\n \"13C, there is shown an example of a watermarked bit stream (in a temporary picture buffer 74) but as it will be transferred to a main buffer 70 to replace the original bitstream for transmission to a decoder.\",\n \"When received at the decoder, the watermarked image will be displayed unless the original image is restored.\",\n \"FIG.\",\n \"16 illustrates an example of an overview of a process according to the invention for restoring (washing) the original image.\",\n \"In step S20, the watermarked bitstream is stored and parsed.\",\n \"In step S22, the encrypted watermark userdata is extracted from the bitstream and stored.\",\n \"Also, any data appended to the end of the bitstream is stored.\",\n \"In step S24, the stored encrypted watermark user data is decrypted to restore the original DC differentials and the AC VLCs.\",\n \"In step S26, restored original DC and AC VLCs replace the DC differentials which represented the watermark.\",\n \"The resultant bitstream is an MPEG2 compressed bitstream but omitting watermarking data.\",\n \"In step S28, the restored bitstream is decompressed in conventional MPEG2 manner.\",\n \"Because all the original coefficients were retained as user data and/or appended to the end of the bitstream, the original bitstream is exactly restorable.\",\n \"The resulting washed file is a bit for bit match to the original unwatermarked file.\",\n \"An example of a process for restoring the original image data is described in more detail in the section “Watermark Wash Code”.\",\n \"Illustrative Implementation (FIGS.\",\n \"17, 18 and 19).\",\n \"Referring to FIG.\",\n \"17, an illustrative watermarking system is shown An uncompressed digital video sequence from a source 50 is applied to a conventional MPEG2 encoder 52.Such encoders are well known.\",\n \"In this example the encoder 52 is a computer running compression software but the encoder could be a special purpose processor.\",\n \"The compressed video sequence is stored as a file in a store 54, e.g.\",\n \"the hard disk of the computer 52.A watermark embedder 56 receives the compressed video sequence from the store 54.In this example the embedder is a computer running embedding software which will be described below but the embedder could be a special purpose processor.\",\n \"The embedding process involves a template such as is shown in FIG.\",\n \"9 and an encryption algorithm which uses at least one encryption key, both of which are needed to restore the original image at the decoder.\",\n \"In this example, the computer has a smart card interface 57 via which the template and the key(s) are stored securely on a smart card 64 or other secure data carrier.\",\n \"Alternatively, the template and key(s) may be transmitted in secure manner across a network or in any other secure manner.\",\n \"The embedder applies the watermark to the compressed bitstream and the watermarked bitstream is transferred to another file store 60 (for example in a studio 62) via a transmission link 58 which may be a wireless link, a network, a data storage device for example a disc or tape, amongst other examples.\",\n \"Referring to FIG.\",\n \"18, a decoder is shown.\",\n \"The watermarked bitstream is taken from the store 60 and applied to a de-embedder also called a “washer” 66 which restores the original bitstream.\",\n \"The washer in this example is a computer running de-embedding software which will be described below but the washer could be a special purpose processor.\",\n \"The washing process involves the template such as is shown in FIG.\",\n \"9 and the encryption algorithm which uses at least one encryption key, both of which are needed to restore the original image at the washer.\",\n \"In this example, the computer has a smart card interface 63 via which the template and the key(s) are received from the smart card 64.The output of the washer is a compressed bitstream which is fed to a conventional MPEG2 decoder 65 for decompression and display on a display 67.Buffers Referring to FIGS.\",\n \"19 (and 13), for the purposes of embedding the watermark data in the compressed bitstream, a file (54 in FIG.\",\n \"12) and three buffers are used.\",\n \"They are: a main buffer 70 which stores the original MPEG2 compliant compressed bitstream from the compressor and which outputs the watermarked bitstream; a watermark data buffer 72 which stores the original DC differentials and AC VLCs; and a temporary picture buffer 74 in which the watermarked bitstream is constructed for transfer to the main buffer.\",\n \"These buffers may be physical buffers or in this example are virtual buffers defined by software.\",\n \"The washing process correspondingly uses three buffers, a main buffer 70, a watermark data buffer 72 and a temporary buffer 74.Watermark Embedding The following sets out, by way of example, a pseudo code implementation of various processes described above.\",\n \"Watermark Embed—Pseudo Code Implementation Example Create a main buffer and a watermark data buffer WHILE pictures still to do Read data for one picture from file into the main buffer Create a temporary picture buffer WHILE NOT at start of displayed image data Parse the main buffer Enable all selected watermarking techniques (flag in the watermark data buffer) WHILE NOT end of picture CALL Process Slice Check that space constraints can be met IF space constraints not met Disable some of the watermarking techniques for this picture (flag in the watermark data buffer) WHILE NOT end of picture CALL Process Slice Determine space available in picture with new watermarked displayed image data IF less than 5 bytes IF original user data present Flag the last block of userdata to indicate it is not watermark data Output the displayed picture data to the main buffer Pad the picture to size using zero bytes ELSE Output a user data start code WHILE there is watermark data AND there is space in the picture to fill Encrypt watermark data Output encrypted watermark data to user data space in the temporary picture buffer Flag the block of user data to indicate it is watermark data Output the displayed picture data to the main buffer IF there is space in the picture to fill Pad the picture to size using zero bytes Write the modified picture from the main buffer to file Write any data remaining in the watermark data buffer to file Process Slice function WHILE in slice IF data is not related to the watermark techniques POP the data from the main buffer PUSH the data to the picture buffer ELSE POP the data from the main buffer PUSH the data to the watermark data buffer Calculate replacement value Encode replacement value PUSH replacement value to the picture buffer Watermark Embed Code By way of explanation of the Watermark Embed code, the MPEG2 compliant bitstream is stored in a file, e.g.\",\n \"54 in FIG.\",\n \"12, the buffers 70, 72 and 74 are created and the data for one picture is written from the file into the main buffer 70 where it is parsed.\",\n \"Next selection of watermarking techniques takes place.\",\n \"The selection of watermarking techniques” are described in more detail below with reference to FIG.\",\n \"22.Initially, all watermarking techniques are enabled.\",\n \"The “process slice function” which will be described in more detail separately is then run.\",\n \"In overview, that function places the original DC differentials and AC VLCs in the watermark data buffer 72; and calculates the new watermark DC differentials (i.e.\",\n \"replacement values) and places the replacement values in the picture buffer 74.A check is then made to ensure that a space constraint can be met.\",\n \"Initially it is assumed that all of a particular set of watermarking techniques is applied to the bitstream.\",\n \"If that does not satisfy the space constraint, then one or more of the techniques are disabled.\",\n \"One space constraint, in this example, is passed if the number of bits freed by removing AC VLCs from the image is greater than the number of bits needed to make changes to the DC differentials.\",\n \"That indicates that the watermarked display image is no greater than the original display image.\",\n \"If it is not met then that indicates that the removal of the AC VLCs has not freed enough space for the watermarking DC differentials.\",\n \"The process slice function is run again with the selected techniques.\",\n \"A step “Determine space available”, in this example, checks the space available in the picture with the new watermark DC differentials in it.\",\n \"If there is less than 5 bytes of spare space then no watermark user data field 20 can be included.\",\n \"If the bitstream includes one or more user data fields 36, the last bit of the last such field 36 is set to zero as shown at 40 in FIG.\",\n \"11C.\",\n \"The displayed picture data which includes the new DC differentials representing the watermark is output from the picture buffer 74 to the main buffer 70 and the picture size is padded to its original size by zero bits; see 22 in FIG.\",\n \"11 or 12.If for example, there is sufficient space in the picture data including the new DC differentials representing the watermark, then a watermark user data field 20 is created in the picture buffer 74 by outputting a user data start code.\",\n \"The original AC VLCs and DC differentials stored in the watermark data buffer 72 are encrypted and placed in the watermark user data field in the temporary picture buffer 74.The last bit of the watermark user data field is set to 1 as shown at 38 in FIG.\",\n \"11B to indicate it contains the original AC VLCs and DC differentials.\",\n \"The contents of the temporary picture buffer 74 are output to the main buffer 70.If there is space to fill in the watermarked displayed image data space 23, the space 23 is filled with zero bits as indicated at 22 in FIG.\",\n \"11C.\",\n \"The data in the main buffer 70 is then written to a file.\",\n \"Any data remaining in the watermark data buffer 72, i.e.\",\n \"the original AC VLCs and DC differentials which are not in the watermark user data, are encrypted and also appended to the end of the bitstream forming the file.\",\n \"Process Slice Function.\",\n \"By way of explanation of this function, data is processed, in this example, in slices as described with reference to FIG.\",\n \"5 above.\",\n \"However that is not essential.\",\n \"It is merely a convenient way for the software to operate.\",\n \"The code POP indicates that data is removed from a buffer.\",\n \"The code PUSH indicates that data is moved into a buffer.\",\n \"Thus the data in the main buffer 70 has been parsed as described above.\",\n \"All data up to the start of the displayed image data is skipped over in the main buffer and left there.\",\n \"The only data which is PUSHed into the picture buffer is the displayed image data.\",\n \"This makes it easier to modify an original user data block.\",\n \"In cases where watermark user data is to be added, it is necessary to insert this in front of the watermarked display image data.\",\n \"Data which is related to watermark data, e.g.\",\n \"the AC VLCs and DC differentials are removed from the main buffer 70 and put into the watermark data buffer 72.The new replacement values of the DC differentials are calculated and encoded and put into the picture buffer 74.Watermark Wash—Pseudo Code Implementation Example Create a main buffer and a watermark data buffer CALL Initialise Watermark Data WHILE pictures still to do Read data for one picture from file into the main buffer Create a temporary picture buffer CALL Get Watermark Data WHILE NOT at start of displayed image data Parse the main buffer Enable watermarking techniques as indicated by the setting of flags in the watermark data WHILE NOT end of picture CALL Process Slice Output the contents of the picture buffer to the main buffer IF there is space in the picture to fill Pad with zeros Write the modified picture from the main buffer to file Initialise Watermark Data Function Isolate watermark user data blocks from the file Decrypt the data Store in a temporary file Get Watermark Data Function IF watermark data buffer is not full AND watermark data still present in temporary file Read watermark data from temporary file into watermark data buffer Process Slice Function (not the Same as the Function Used to Embed the Watermark) WHILE in slice IF data is not related to the watermark techniques POP the data from the main buffer PUSH the data to the picture buffer ELSE POP the data from the main buffer POP original data from the watermark data buffer PUSH original value to the picture buffer Watermark Wash Code The watermark wash code restores the original image data.\",\n \"By way of explanation, a main buffer 70 and a watermark data buffer 72 and a temporary buffer 74 are created at the decoder 66.Also a temporary file is created by a function “Initialise Watermark Data function”.\",\n \"The initialise watermark data function isolates the watermark user data blocks from the file, i.e.\",\n \"the encrypted original AC and DC differentials, decrypts them (using the key from the smart card 64) and stores the decrypted AC VLCs and DC differentials in the temporary file.\",\n \"If there is picture data which has not been washed, that data is read from the temporary file into the main buffer.\",\n \"The temporary buffer 74 is created at this stage.\",\n \"A function Get Watermark Data determines whether the watermark data buffer 72 is not full AND watermark data is still present in the temporary file.\",\n \"If so, then watermark data is read from the temporary file into the watermark data buffer.\",\n \"This ensures that there is always at least enough data in the watermark data buffer to wash the current picture.\",\n \"“Enable watermarking techniques .\",\n \".\",\n \".\",\n \"” reads the 1 bit flag from the watermark data to determine if the picture was successfully watermarked using all selected techniques.\",\n \"If it was not, then additional flag bits are read to determine which techniques were used.\",\n \"CALL process slice operates as follows: POP and PUSH indicate the same operations as given above.\",\n \"Data in the main buffer has been parsed.\",\n \"Data in the main buffer which is not related to watermarking is removed and placed in the picture buffer 74.Watermark data (i.e.\",\n \"the DC differentials which represent the watermark) is removed from the main buffer 70.The decrypted original AC VLCs and DC differentials in the watermark data buffer 72 are removed therefrom and put into the picture buffer 74.Further steps transfer the data from the picture buffer to the main buffer Finally, the restored picture is written from the main buffer to a file.\",\n \"It will be appreciated that the process described above requires the template stored in the data carrier 64 to determine which blocks contain watermark data and which do not.\",\n \"Also the key(s) needed for decryption are derived from the data carrier 64.Motion Vectors which have been modified as described in the section below “Fourth Example—Motion Vectors” are also restored to their original values in the same way.\",\n \"SECOND EXAMPLE Non-Intra DC Changes In the first example, the watermark is applied by replacing the original DC differential values of the DC coefficients in template blocks in Intra frames with new values representing the watermark.\",\n \"No other coefficients are replaced.\",\n \"The original values are stored in the watermark user data pspace and/or at the end of the bitstream.\",\n \"In a second example of the invention, in addition to replacing the intra frame DC coefficients, the DC coefficients in template blocks in non-intra frames (that is P and B frames) are replaced by new values and the original values stored in the watermark user data space and/or at the end of the bitstream.\",\n \"In non-intra frames, DC coefficients are coded as the difference between the current DC coefficient and the desired coefficient is coded since the magnitude of the change is usually small.\",\n \"However, there are instances where the magnitude might be much larger and this may cause the DC value of a watermarked block to clip at the limits of the range.\",\n \"To avoid this the magnitude of the DC coefficient can be changed to the minimum value of 1.\",\n \"(Changing the value to zero would completely eliminate the VLC and alter the run-length of the following code).\",\n \"The original VLC is stored in the watermark data buffer 72 and the new code put in its place.\",\n \"The VLC representing a run-length of zero and a level change of ±1 is the shortest VLC in the coding table so this process guarantees never to increase the number of bits required to code the picture.\",\n \"It can therefore always be performed whenever intra DC changes are also being performed.\",\n \"The sign of the level change is set to be positive when the DC change is to black and negative when the change is to white.\",\n \"This prevents clipping of the DC from occurring.\",\n \"Referring to FIG.\",\n \"20, non-intra frames are processed as follows.\",\n \"Step S30 determines the frame type.\",\n \"If the frame is a non-intra frame the process proceeds to step S32, otherwise if the frame is an intra-frame the process proceeds as set out in FIG.\",\n \"10.In step S34, the codes representing DC coefficients (which are difference values as described above) in template blocks are removed and stored in the watermark data buffer 72 (see FIG.\",\n \"12B).\",\n \"In step S36, new change values of +/−1 are placed in the bitstream in place of the removed coefficients.\",\n \"In steps S36 and S38, the bitstream is reformed in the temporary buffer 74 (see FIG.\",\n \"12C), the removed coefficient values being placed in the watermark userdata space 20 if space is available and otherwise in step S40 appended to the end of the bitstream.\",\n \"The original DC coefficients are encrypted as described above.\",\n \"AC VLCs are not changed in non-intra frames.\",\n \"THIRD EXAMPLE Frame Pictures, Changing DCT type, FIG.\",\n \"21 The first and second examples may be modified as follows.\",\n \"There are two types of picture: field and frame.\",\n \"In field pictures only one type of DCT block occurs.\",\n \"In this, third, example, we are concerned only with frame pictures.\",\n \"In frame pictures two types of DCT blocks can occur; field and frame.\",\n \"The field DCT is used when the video source is interlaced because it improves the correlation between adjacent rows and consequently increases the proportion of the signal power in the lower frequency components of the DCT.\",\n \"However, it is not used exclusively for this purpose.\",\n \"In a macroblock all the blocks have the same type.\",\n \"Referring to FIG.\",\n \"21A, if a DC change is made in a frame picture to one DCT block B1 but not to the DCT block either directly vertically above or below it in the same macroblock MB and the field type is being used for the DCT blocks of that macroblock then the change will result in stripes across the entire height of the macroblock.\",\n \"This is not desirable as it distorts the watermark and if the watermark is in the form of alpha-numerics they may become harder to read.\",\n \"The appearance of stripes is entirely dependent on the encoding of the content and in particular the proportion of macroblocks coded with the field DCT.\",\n \"In accordance with an example of the present invention, to solve this problem the DCT type of any macroblock in a frame picture, which contains at least one DCT block to be watermarked, is set to frame.\",\n \"This involves storing the original DCT type code in the watermark data buffer 72 (FIG.\",\n \"13) and replacing it with the code for a frame DCT.\",\n \"This does not to affect the number of bits required by the picture since the DCT type code is always a 1-bit value.\",\n \"This change can therefore always be made whenever the DC watermark is used.\",\n \"As with motion vector changes, the DCT type operates at the macroblock level so any change will affect all DCT blocks within the macroblock regardless of whether they are watermarked or not.\",\n \"In this case the side-effect that occurs is a re-ordering of the rows within the macroblock.\",\n \"This may result in an obviously, visibly, incorrect macroblock but the appearance of the macroblock may not be objectionable.\",\n \"FIG.\",\n \"21B illustrates an example method to process the DCT type (including, where appropriate, changing from field to frame DCT and reversing this change).\",\n \"In FIG.\",\n \"21B: “picture_structure==FRAME” detects Frame pictures; “&&” is logical AND “frame_pred_frame_dct ==0” detects that the macroblocks have the DCT type specified on an individual basis rather than being universally set to frame DCT type for this picture; ∥ is logical OR; and “macroblock intra∥macroblock_pattern” detects intra-encoded macroblocks and non-intra blocks which include coded DCT blocks.\",\n \"Intra-encoded macroblocks have all DCT blocks coded.\",\n \"It is necessary to detect this because only coded blocks have a DCT type.\",\n \"DCT type (dct_type) is denoted by a 1 bit code: 1=field type; 0=frame type.\",\n \"The original bitstream is stored in a main buffer 70 and the codes which are removed are temporarily stored in a watermark data buffer 72 (see FIG.\",\n \"13).\",\n \"The process starts at step S70 and proceeds to step S71 and S72 which detect macroblocks which have a DCT type.\",\n \"If a macroblock has no DCT type the process ends at step END.\",\n \"If a macroblock has a DCT type then the 1 bit code representing DCT type is removed (POPped) from the main buffer 70 in step S73.Step S74 determines that DC changes are enabled (see the section “selective techniques” below) and the macroblock is within the template.\",\n \"If yes, step S75 determines whether a watermark is being embedded in the bitstream or a watermarked bitstream is being restored to its original form (washed).\",\n \"If the bitstream is being watermarked, then in step S76, the dct_type code (that was removed from the main buffer in S73) is placed in (PUSHed into) the watermark data buffer 72 (see FIG.\",\n \"13).\",\n \"The value of the dct_type code is then changed to frame type (=0) in step S77 and in step S79 the new code is PUSHed into the temporary buffer 74 (see FIG.\",\n \"13).\",\n \"If the NO route is taken from step S74 the POPped dct_type code is PUSHed into the temporary buffer in step S79.In the reverse process steps S70 to S74 proceed as described above.\",\n \"Then at step S75 it is determined that the watermarking process is being reversed (washing) and at step S78 the dct_type code for the watermarked stream is replaced by the original one that is extracted from the watermark data buffer 72.In step S79, the dct_type code is then PUSHed into the temporary buffer 74 as before.\",\n \"At the end of the process shown in FIG.\",\n \"21B, the temporary buffer 74 contains the new dct_type code and the watermark data buffer 72 contains the original codes.\",\n \"The original dct_type codes from buffer 72 are transferred to the watermark user data spaces 20 or appended to the end of the bitstream.\",\n \"The new dct_type codes are placed in the bitstream in place of the original codes.\",\n \"That is done in the same way as described above in the section “Watermark Embed Code”.\",\n \"Selective Techniques As described above, a picture is watermarked by: a) First Example—removing all codes representing original DC and AC coefficients from template blocks in intra frames and replacing the DC coefficients by new values representing a watermark; b) Second Example—optionally, in addition removing codes representing original DC coefficients in non-intra frames and replacing them with new values; c) Optionally, in addition, replacing original motion vector values of template blocks with zero values.\",\n \"More generally three techniques may be used: 1.Enabling change of DC coefficients in template blocks; 2.Enabling removal of AC coefficients from template blocks; and; 3.Enabling change of Motion Vector values in template blocks.\",\n \"Usually, a picture watermarked using the example techniques described here will require fewer bits to encode than the original picture.\",\n \"In the first example this is due to the complete removal of AC coefficients but the coding of a large number of zero differential values for both DC and motion vectors also makes a significant contribution.\",\n \"However, there are instances where, to embed the watermark completely, the number of bits inserted into a picture ends up being greater than the number of bits removed from that picture.\",\n \"For example, this can easily occur with a completely black frame, commonly encountered in video sequences.\",\n \"If this occurs, then in order to satisfy the constraint that the number of bits used to represent the picture must remain the same it becomes necessary to disable some of the techniques used to watermark the picture until the constraint can be met.\",\n \"In examples of the invention, the selection is made on a per-picture basis so it is necessary to store additional information to facilitate correct washing of the picture.\",\n \"Flag bits are inserted to indicate which techniques are enabled for the current picture.\",\n \"The system used at present is shown in Table A.\",\n \"It easily adapts to the addition of further techniques and is also efficient regardless of the selected subset of techniques chosen from the full set of techniques available.\",\n \"Note that the choice of the first flag bit is designed to prevent the occurrence of MPEG-2 start codes within the watermarked data.\",\n \"TABLE A Selective technique flag bits Whenever possible the intra DC change and AC removal techniques should both be applied as a minimum since this gives good visibility and robustness.\",\n \"If intra DC changes are applied then non-intra DC and DCT type changes can also be used since they are guaranteed not to increase the number of bits required by the picture.\",\n \"Changes to the motion vectors improve the visual quality and integrity of the watermark but are not essential.\",\n \"FIG.\",\n \"22 is a flow diagram illustrating the selection, by way of example, of the three watermarking techniques: 1.Enabling change of DC coefficients in template blocks; 2.Enabling removal of AC coefficients from template blocks; and; 3.Enabling change of Motion Vector values in template blocks.\",\n \"The following points should be noted: AC bits freed is always non-negative.\",\n \"AC removal can always be performed and will never result in an increase in picture size.\",\n \"Bits for DC changes and Bits for MV changes may be positive or negative.\",\n \"If the value is positive then the number of bits needed to perform the technique is greater than the number of bits originally present.\",\n \"A sufficient number of bits must therefore be freed using another technique (e.g.\",\n \"AC removal) if the changes are to be performed on the current picture.\",\n \"If the value is negative the technique requires fewer bits than originally present so some bits have actually been freed and the changes can always be performed.\",\n \"DC changes are given priority over MV changes whenever the number of bits freed is insufficient to compensate for the bits needed to perform both techniques Referring to FIG.\",\n \"22, the process starts at step S80 and proceeds to step S81 at which initially all three techniques are enabled.\",\n \"In step S82 a picture (that is a frame or field) is processed to apply all of the three enabled techniques.\",\n \"Step S83 sets a reprocess flag to true to indicate that initially it is assumed that the picture can be reprocessed using all three techniques.\",\n \"In overview, a picture is processed in one pass using all three techniques to apply a watermark and the result is analysed.\",\n \"If the result is not successful, the picture is reprocessed in a second pass with one or more of the techniques disabled.\",\n \"AC Bits Freed Allowing All Three Techniques Step S84 determines whether removing the AC coefficients removes any data thus freeing bits for changing the DC coefficients.\",\n \"(For a black picture there may be no AC coefficients).\",\n \"If the number of bits freed exceeds zero then step S85 determines whether the number of AC bits freed exceeds the sum of bits for DC changes and bits for MV (Motion Vector) changes.\",\n \"The number of bits for DC changes is the difference between the number of bits freed by removing the original DC codes and the number of bits used by the new DC codes.\",\n \"Likewise the number of bits for MV changes is the difference between the number of bits freed by removing the original MV codes and the number of bits used by the new MV codes.\",\n \"If the number of AC bits freed is NOT less than the sum of the bits for DC changes ad the bits for MV changes (i.e.\",\n \"sufficient AC bits are freed to allow both DC and MV changes) then all three techniques can be used and so there is no requirement to reprocess the picture.\",\n \"Thus the reprocess flag is changed to false in step S90.Step S91 determines whether the reprocess flag is true or false.\",\n \"In this instance it is false so the process ends at the END step.\",\n \"Step S84—no AC Bits Freed This may occur on a black picture which thus has no AC coefficients and so it is not possible to embed any watermark.\",\n \"That is not a problem because a black picture has no content to protect.\",\n \"If in step S84 it is determined that no AC bits are freed then the process proceeds to step S93 which disables the AC removal technique.\",\n \"Step S94 tests whether the bits for DC change is greater than zero.\",\n \"If it is then the DC change technique is disabled in step S95.If there are no bits for DC change or step S95 has occurred the process proceeds to step S96 which test whether the number of bits for MV change is greater than zero.\",\n \"If it is then in step S97 the MV change technique is disabled.\",\n \"In summary if no AC bits are freed, the DC and MV changes are disabled or there are no DC and MV change bits.\",\n \"In either case the reprocess flag is still true and so the picture is processed (again) at step S92.Note that whilst ‘AC bits freed ’ is always non-negative, ‘bits for DC changes’ and ‘Bits for MV changes’ may be positive or negative.\",\n \"If there is no AC data to remove, then DC and MV changes can only be performed if the replacement data has fewer bits than the original data: i.e.\",\n \"‘Bits for DC changes <=0 and similarly for MV changes.\",\n \"It is therefore possible (but unlikely) to have DC and/or MV changes enabled with AC removal disabled.\",\n \"It is always necessary to reprocess when any of the techniques is disabled since the original assumption is that all three techniques are enabled so the flag data must be changed and the first pass watermarked frame is overwritten by the second pass watermarked frame.\",\n \"The lack of AC VLC data means that no space can be freed in the picture data to allow changes in the differential DC values that increase the number of bits.\",\n \"In such instances it may not be possible to embed the watermark but this is not a problem since there is no content to protect.\",\n \"However, a decoder which removes the watermark herein referred to as a watermark washer must be told where these instances have occurred so that it does not attempt to remove the watermark from the frame.\",\n \"Additional flag bits are used to indicate this.\",\n \"AC Bits Less than the Sum of Bits for DC and MV Changes Step S85.If there are insufficient bits freed to do both DC and MV changes then the process proceeds to step S86.In this case, step S86 tests whether the number of AC bits freed is sufficient to do the DC changes.\",\n \"If there are sufficient bits to do the DC changes (NO output of S86) then step S97 occurs and MV changes are disabled.\",\n \"The process then proceeds to step S91.Because the reprocess flag is true, but MV changes are disabled, then the picture is processed anew with MV changes disabled and DC changes enabled in step S92.If step S86 determines that there are insufficient bits freed to do the DC changes (YES output of S86), then the process proceeds to step S87 which tests whether the number of AC bits freed is sufficient to do the MV changes.\",\n \"If there are sufficient bits to do the MV changes (NO output of S87) then step S88 disables DC changes.\",\n \"The process then proceeds to step S91.Because the reprocess flag is true, but DC changes are disabled, then the picture is processed anew with DC changes disabled and MV changes enabled in step S92.If step S87 determines that there are insufficient bits to do the MV changes (YES output of S87) then step S890 disables both DC and MV changes and the picture is reprocessed via steps S91 and S92 with only the AC removal technique enabled.\",\n \"In a modification of the process of FIG.\",\n \"22, AC removal is permanently enabled because it has been found that AC removal never causes problems.\",\n \"In FIG.\",\n \"22, step S93 is then removed and a fig bit for AC removal is not required.\",\n \"FOURTH EXAMPLE Motion Vectors—FIGS.\",\n \"23 to 25 Setting Motion Vectors to Zero Referring to FIG.\",\n \"23A, motion vectors are associated with macroblocks (shown by bold squares MB) rather than DCT blocks (shown by lighter squares B).\",\n \"Thus a motion vector template as indicated by light shading may be defined based on the watermark template shown by dark shading.\",\n \"Even if, as assumed above in the first example, only I frames contain watermark data, motion vectors may undesirably distort the watermark.\",\n \"It is preferable but not essential to determine whether any macroblock in a P and or B frame falls within the motion vector template.\",\n \"If it does, then the value of any motion vector of that macroblock is set to zero.\",\n \"Motion vectors are differentially encoded so the differential value is set to represent a value of zero for the motion vector.\",\n \"(That is, Motion Vectors are processed in the same way as described above for the DC differentials).\",\n \"As described above with reference to the DC differentials, the original codes representing the Motion Vectors of DCT macroblocks, which include DCT blocks within the template, are removed from the bitstream and replaced by modified codes which in this case represent Motion Vectors of zero value.\",\n \"The original Motion Vector codes are placed in the watermark user data spaces and/or appended to the end of the bitstream.\",\n \"Thus referring to FIGS.\",\n \"13 and 24:— The bitstream is stored in the main buffer 70.In step S100, it is determined whether in a non-intra frame a macroblock falls within the motion vector template.\",\n \"If it does then the motion vector code associated with that macroblock is removed from the bitstream (S102) in the main buffer 70 and stored (S104) in the watermark data buffer 72.A new motion vector code representing zero value for motion vector is placed in the bitstream in place of the original value in the temporary buffer 74 (S106) and the original motion vector codes are placed in the watermark user data space 20 in the temporary buffer 74 (step S108) and/or appended to the end of the MPEG compliant bitstream (S110).\",\n \"The original motion vector codes are encrypted before being placed in the user data space or appended to the end of the bitstream as described above with reference to FIG.\",\n \"15.Motion Vectors may be set to zero as described in this section in any of the examples given above.\",\n \"Whilst this reduces distortion of the watermark, the watermark may still be distorted especially at the edges.\",\n \"The next section headed “Borders” describes an illustrative method of further reducing distortion of the watermark.\",\n \"Borders FIG.\",\n \"23B Referring to FIG.\",\n \"23B, to further reduce distortion of the watermark, the motion vector template is extended to include macroblocks which do not include watermark template blocks but which border such blocks.\",\n \"The motion vector codes of all the macroblocks in the extended motion vector template are processed as described above with reference to FIG.\",\n \"24.This reduces the distortion of the watermark as defined by the watermark template but because the motion vectors of the macroblocks bordering the watermark template, which do not include any watermark blocks, the picture is distorted in the bordering macroblocks.\",\n \"It is a matter of subjective judgement whether the reduced distortion of the watermark is outweighed by the increased distortion of the picture.\",\n \"Changing the Values of Motion Vector Codes, FIG.\",\n \"25 Motion Vectors are differentially encoded, and are recoded, in the same way as DC coefficients as described above in the section headed “Differential DC recoding”.\",\n \"P frames have forward prediction motion vectors but B frames have two types of motion vectors: forward prediction and backward prediction.\",\n \"The two types are processed separately as will be described with reference to FIG.\",\n \"25.FIG.\",\n \"25A shows a much simplified motion vector template of three macroblocks MB between other macroblocks O which are not part of the watermark and its border.\",\n \"FIG.\",\n \"25B shows the prediction types of the macroblocks.\",\n \"F indicates forward and B indicates backward.\",\n \"Some macroblocks FB are both forward and backward predictive.\",\n \"Referring to FIGS.\",\n \"25 C to E, the numbers shown in those Figures are for explanation only and do not purport to represent realistic values for motion vectors.\",\n \"FIG.\",\n \"25C shows non-differential values for forward predictive motion vectors.\",\n \"The corresponding differential values are shown in FIG.\",\n \"25D.\",\n \"It will be noted that the backward predictive motion vectors are ignored.\",\n \"When the motion vectors in the three template macroblocks are set to zero, the differentials of those blocks are recalculated so the effective values within the template are effectively zero but the effective values of neighbouring blocks outside the template are not changed.\",\n \"That results in the differential value of the macroblock at the leading edge of the template denoted “Entry” changing relative to its preceding macroblock to provide effectively zero motion vector value and the differential value of the trailing edge macroblock denoted “Exit” changing to maintain the original value of the next bordering macroblock.\",\n \"It will be appreciated that the forward predictive motion vectors values are in a sequence of differential values which are independent of the backward predictive values and that the Entry and Exit macroblocks of the two sequences do not necessarily coincide.\",\n \"It will also be appreciated that the original differential value of the motion vector of the macroblock bordering the exit from the templates does not need to be stored as described above for block 65 in the section “Differential DC Recoding” but it is preferred to store it.\",\n \"Templates—FIGS.\",\n \"26 to 28 As discussed above, there are two types of picture: field and frame.\",\n \"A video sequence may contain: only field pictures; only frame pictures; or both field and frame pictures.\",\n \"Sequence of Frame Pictures Only Consider a sequence of only frame pictures.\",\n \"A template is defined to a resolution of one DCT block horizontally and vertically (8 pixels by 8 pixels).\",\n \"For the example frame picture shown in FIG.\",\n \"26A, having 720 pixels per line and 576 lines, the required template size is 90 blocks by 72 blocks.\",\n \"A template in the form of an X is defined to the resolution of one block in FIG.\",\n \"26B.\",\n \"The resulting watermarked picture shown in FIG.\",\n \"26C has a displayed watermark resolution of one block of 8×8 pixels.\",\n \"Sequence of Field Pictures Only Consider a sequence of only field pictures.\",\n \"Referring to FIGS.\",\n \"27 A1 and A2, two fields each of 720×288 pixels are interlaced to define an interlaced frame of 720×576 pixels.\",\n \"In a field picture, a DCT block still corresponds to an 8×8 pixel block even though the number of lines is half that of a frame picture.\",\n \"A macroblock is 2×2 DCT blocks as in a frame picture.\",\n \"A template (X in FIG.\",\n \"27B) for a field picture is defined to the resolution of one DCT block.\",\n \"The same template is used for both fields to prevent the occurrence of stripes.\",\n \"No special adjustment of the template for each field is required because block positions are fixed relative to the start of a field.\",\n \"Referring to FIG.\",\n \"28 in respect of one macroblock as shown in FIG.\",\n \"28C a template indicates one block shown in solid black is to be watermarked.\",\n \"In the interleaved frame, two macroblocks from the respective two fields are interleaved to form a composite of two DCT blocks horizontally by four DCT blocks vertically as shown in FIGS.\",\n \"28A and 28D.\",\n \"The composite consists of a macroblock B1 from field one and a macroblock B2 from field two.\",\n \"The lines of the interlaced frame are indicated by F1 for lines from field one and F2 for lines from field two.\",\n \"The watermark X as displayed has a resolution of 8 pixels horizontally by 16 pixels vertically as shown in FIG.\",\n \"28D even though the template has a resolution of 8 pixels by 8 pixels as shown in FIG.\",\n \"28C.\",\n \"Sequence of Field and Frame Pictures Consider a video sequence having at least one field picture and at least one frame picture.\",\n \"The template, X in FIG.\",\n \"27B, is designed for the one or more field pictures and has a resolution of one DCT block.\",\n \"That results in a displayed watermark having a resolution of one block horizontally by two blocks vertically.\",\n \"The watermark in the one or more frame pictures needs to be consistent with that of the field pictures; this can be ensured by automatically deriving the template for frame pictures from the template used for field pictures.\",\n \"The simplest way to achieve this is to double up every row of the field template.\",\n \"The frame template then has an effective resolution of one block horizontally by two blocks vertically to ensure the displayed watermark in the frame pictures matches that of the field pictures.\",\n \"In practice a video sequence is examined to determine the type(s) of picture in it.\",\n \"For field-only and frame-only sequences, a suitable template for that type of picture is used.\",\n \"For a sequence having both types of picture, a template suitable for field pictures is chosen and used for the field pictures in the sequence.\",\n \"A template for use with the frame pictures is then derived from the field template to ensure a consistent appearance to the watermark.\",\n \"The templates may be bit-maps stored in one or more stores.\",\n \"Modifications Packetised Bit Stream Where the original sequence is an MPEG-2 packetised stream (either packetised video sequence or audio/video multiplex) the video sequence end code is not required provided that the watermarked video sequence is repacketised so that the residual watermark data is placed after the Packetised Stream end code.\",\n \"To simplify the embedding of packetised streams (audio/video multiplex), the stream can be pre-processed to extract the video sequence.\",\n \"The video sequence can then be embedded separately as described above.\",\n \"The embedded video sequence can then be repacketised with the audio using the existing packetised stream file as a template, allowing reuse of all packet information and preserving all timing (a useful side-effect of maintaining the picture data size).\",\n \"This additional stage of processing is not described in the psuedo code above since it is not essential to the operation of the embedding process.\",\n \"Varying Compression Ratio The examples described above assume the video is compressed for minimum compression to maximise picture quality before the watermark data is embedded in the compressed bitstream.\",\n \"Thus the watermark user data fields are unlikely to contain all the encrypted original coefficients and the residual data is appended to the end of the file.\",\n \"In a version of the invention, which is not currently preferred, an estimate of the data space needed to contain the watermarked data is provided before compression is carried out and the compression controlled to provide sufficient spare space to accommodate all the original coefficients in watermark user data fields and the new DC differentials in the displayed image data fields.\",\n \"Appending the Encrypted Original AC VLCs and DC Differentials.\",\n \"In the examples described above the original AC VLCs and DC differentials in excess of those stored in the watermark user data fields are appended to the end of the file.\",\n \"However, they could be placed at the beginning of the file.\",\n \"In a currently preferred implementation the number of residual bytes appended to the end of the file is counted.\",\n \"The count is placed after the residual data to enable the start of the residual data to be easily accessed.\",\n \"Alternative Method of Storing Original AC VLCs and DC Differentials in User Data Fields In the foregoing description, the original AC VLCs and DC differentials are stored in the user data fields 20 of the compressed bitstream.\",\n \"As shown in FIG.\",\n \"14, in the example of frames P1 and P2, some data of frame P1 is stored in the user data space 20 of frame P1 and any data in excess of that is stored in the user data space of frame P2.That process continues throughout the bitstream.\",\n \"Any residual data is appended to the end of the bitstream.\",\n \"In a modification of that process, as shown in FIG.\",\n \"29, original AC VLCs and DC differentials of frame P1 are stored in the user data space 20 of frame P1.For every frame there is produced a residual data field 291 comprising the number of bytes B of encrypted AC VLCs and DC differentials which are not included in the user data space of that frame together with the encrypted AC VLCs and DC differentials.\",\n \"If there are no residual AC VLCs and DC differentials, the number B for that frame is zero.\",\n \"Thus each frame has in the user data spaces 20 thereof only data originating from that frame and the residual data is stored elsewhere for example in a buffer 290 in FIG.\",\n \"29.The residual data, i.e.\",\n \"contents of the buffer 290 in this example, may be appended to an end of the MPEG bitstream as shown in FIG.\",\n \"30.As indicated at 292 it may be appended to the end at the start of the bitstream or as indicated at 293 it may be appended to the end at the finish of the bitstream.\",\n \"The codes B representing the number of bits of user data are chosen so that they do not represent or can be confused with other codes used in MPEG bitstreams, especially MPEG start codes.\",\n \"Files As described above, any residual data comprising the codes B and the encrypted residual AC VLCs and DC differentials is appended to an end of the bitstream.\",\n \"In a modification, that residual data is stored in a file separate from the bitstream.\",\n \"The file may be appended to an end of the bitstream as shown in FIG.\",\n \"30 at the beginning 292 of the bitstream or at the finish 293.As shown in FIG.\",\n \"31, the file may be stored in a transmission medium 294 for example a data carrier or a communications channel separate from the bitstream.\",\n \"The data carrier may be a floppy disc or tape or other storage medium.\",\n \"The file is associated with the bitstream.\",\n \"That may be done by appending it to the bitstream as shown in FIG.\",\n \"30.If the file is stored separately as shown in FIG.\",\n \"31 it may be associated with the bitstream by use of for example an identifier which links the file to the bitstream.\",\n \"An example of such an identifier is a UMID.\",\n \"Washing To wash the watermark from the bitstream of FIG.\",\n \"29, the bitstream P1 to Pn is stored and parsed.\",\n \"The residual data comprising the codes B and the AC VLCs and DC differentials is retrieved from the appended data 292 or 293 in the parsing process or from the file 292, 293 or 294 and stored.\",\n \"For each frame, the data in the user data field 20 is decrypted and used to replace the modification data in the bitstream.\",\n \"The number of bytes B is read for that field.\",\n \"If it is greater than zero, then the residual data for that field is decrypted and used to replace the remaining modification data of that frame.\",\n \"In the example of an MPEG bitstream the frames are not numbered nor are the residual data fields 291.Such numbering is not needed: every frame and its associated residual data field are processed in succession.\",\n \"Such numbering could be provided in other examples of the invention.\",\n \"Selecting Watermarking Techniques The foregoing description describes the selection of three watermarking techniques.\",\n \"However selection could be made from many techniques including: a) replacement of DC differentials by modified DC differentials and removal of the original DC differentials; b) removal of AC VLCs to make space for the original DC differentials in the bitstream; c) placing removed coefficients in user data blocks; d) placing removed coefficients at the end of the bitstream; e) modification of motion vectors for watermarking; f) modification of colour information for watermarking; and g) placing coefficients in spare space in P and/or B frames.\",\n \"The techniques may be used separately or together.\",\n \"For example it may be possible in some circumstances to replace only the DC differentials with modified coefficients without removing AC VLCs.\",\n \"Typically, but not always, modification of coefficients and motion vectors for watermarking increases the number of bits.\",\n \"The techniques used may be selected depending on the effect they have on the bitrate.\",\n \"If a particular set of techniques results in too many bits (e.g.\",\n \"a) plus c)) then a different set of techniques may be selected.\",\n \"The techniques used are indicated in the watermarked bit stream by flags which identify the techniques so that when the watermarked bitstream is applied to a washer for removing the watermark the washer is supplied with the information needed to correctly remove the watermark.\",\n \"Applications The examples of the invention described above can be applied in the following contexts.\",\n \"Content Brokering The visibly watermarked files produced using the technique described allow preview of content using a standard MPEG-2 player (the watermarked file is a valid MPEG-2 stream) there is no need for a bespoke player.\",\n \"The watermarked file contains all information needed to reverse the process (apart from the encryption key and the template file).\",\n \"It is therefore only necessary to distribute/transmit one copy of the content (as compared with the conventional procedure of distributing a protected copy of the sequence followed by an unprotected copy when the buyer has purchased the content).\",\n \"There are a number of forms that this distribution can take: The watermarked content could be stored on a secure server but the encryption of the watermark data within the file could be performed at the moment a copy of the file is requested.\",\n \"The client can enter their details which are then stored on the secure server with a copy of the encryption key used.\",\n \"If the client then wishes to purchase the content the encryption key can be retrieved from the secure server using the client's details as an index.\",\n \"This means that one client's key cannot be used to wash another client's copy of the watermarked content.\",\n \"Of course, it would still be possible for the client with the key to provide their copy of the content (either watermarked or washed) to someone else.\",\n \"Once the client has purchased the content there is the option of either washing the file to produce a file without the visible watermark or to leave the local copy of the file watermarked and to wash it in real-time as required.\",\n \"The former case is likely to be acceptable since it is no different to the current model.\",\n \"Pay-Per-View Broadcast The nature of the watermarking process makes it possible to develop a real-time washer.\",\n \"This means that protected content can be streamed to the viewer for preview.\",\n \"If the viewer subsequently decides to pay for the content they can be given a key to enter into their set-top box (for example) and the content is washed locally as it arrives—there is no need to change the data being transmitted.\",\n \"In order to facilitate real-time washing it may be necessary to download all the encrypted watermark information to the set-top box first.\",\n \"Alternatively, the content compression process can be modified to incorporate padding in the picture frames sufficient to compensate for the additional watermark data.\",\n \"With a given template design calculations can be made to determine the worst case increase that may need to be compensated.\",\n \"The invention may be implemented by hardware or software or a combination of both.\",\n \"The invention provides a computer program which when run on a computer implements the methods described above.\",\n \"The invention also provides a store storing such a program.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10474013"}}},{"rowIdx":780,"cells":{"text":{"kind":"list like","value":[["Dual core photonic crystal fibers(pcf) with special dispersion properties","An optical fibre for transmitting light, said optical fibre having an axial direction and a cross section perpendicular to said axial direction, said optical fibre comprising: (1) a first core region comprising a first core material having a refractive index Nco,1; (2) a microstructured first cladding region surrounding the first core region, said first cladding region comprising a first cladding material and a plurality of spaced apart first cladding features or elements that are elongated in the fibre axial direction and disposed in the first cladding material, said first cladding material having a refractive index Ncl,1 and each said first cladding feature or element having a refractive index being lower than Ncl,1, whereby a resultant geometrical index Nge,cl, 1?","of the first cladding region is lowered compared to Ncl,1; (3) a second core region surrounding said first cladding region, said second core region comprising a second core material having a refractive index Nco,2, and (4) a second cladding region surrounding the second core region, said second cladding region comprising a second cladding material having a refractive index Ncl,2, wherein the first core material, the first cladding material and the first cladding features, the second core material, and the second cladding material are selected and arranged so that Nco,1>Nge,cl,1, Nco,2>Nge,cl,1, and Nco,2>Ncl,2."],["1-103.","(canceled) 104.An optical fiber for transmitting light, said optical fiber having an axial direction and a cross section perpendicular to said axial direction, said optical fiber comprising: (1) a first core region comprising a first core material having a refractive index Nco,1, (2) a microstructured first cladding region surrounding the first core region, said first cladding region comprising a first cladding material and a plurality of spaced apart first cladding features or elements that are elongated in the fiber axial direction and disposed in the first cladding material, said first cladding material having a refractive index Ncl,1 and each said first cladding feature or element having a refractive index being lower than Ncl,1, whereby a resultant geometrical index Nge,cl,1 of the first cladding region is lowered compared to Ncl,1, (3) a second core region surrounding said first cladding region, said second core region comprising a second core material having a refractive index Nco,2, and (4) a second cladding region surrounding the second core region, said second cladding region comprising a second cladding material having a refractive index Ncl,2, wherein the first core material, the first cladding material and the first cladding features, the second core material, and the second cladding material are selected and arranged so that Nco,1 >Nge,cl,1, Nco,2 >Nge,cl,1, and Nco,2>Ncl,2.105.The optical fiber according to claim 104 wherein the second cladding region is homogeneous.","106.The optical fiber according to claim 104 wherein the second core region is micro-structured having a plurality of spaced apart second core features or elements that are elongated in the fiber axial direction and disposed in the second core material, each said second core feature or element having refractive index, Nco,2 being higher than a refractive index, Nco,back,2, of the second core material, whereby a resultant geometrical index Nge,co,2 of the second core region is higher compared to Nco,back,2.107.The optical fiber according to claim 104 wherein Ncl,2 is equal to or lower than Nco,back,2.108.The optical fiber according to claim 104 wherein Ncl,2 is equal to or lower than Ncl,1.109.The optical fiber according to claim 104 wherein the second cladding region is micro-structured having a plurality of spaced apart second cladding features or elements that are elongated in the fiber axial direction and disposed in the second cladding material, each said second cladding feature or element having a refractive index being lower than Ncl,2, whereby a resultant geometrical index Nge,cl,2 of the second cladding region is lowered compared to Ncl,2.110.The optical fiber according to claim 104 wherein the first core region is a solid core region.","111.The optical fiber according to claim 104 wherein Nco,1 is larger than Nco,2.112.The optical fiber according to claim 104 wherein Nco,1 is identical to Nco,2.113.The optical fiber according to claim 104 wherein Nco,1 is lower than Nco,2.114.The optical fiber according to claim 104 wherein said second core region is a homogeneous region.","115.The optical fiber according to claim 104 wherein the second core region is micro-structured having a plurality of spaced apart second core features or elements that are elongated in the fiber axial direction and disposed in the second core material, each said second core feature or element having a refractive index being lower than Nco,2 whereby a resultant geometrical index Nge,co,2 of the second core region is lowered compared to Nco,2, and wherein Nge,co,2>Nge,cl,1, and Nge,co,2>Nge,cl,2.116.The optical fiber according to claim 106 wherein the second core region further comprises low index second core features.","117.The optical fiber according to claim 104 wherein the first cladding features or elements are positioned equidistantly with respect to a center of the fiber.","118.The optical fiber according to claim 117 wherein the first cladding features or elements are of similar size.","119.The optical fiber according to claim 106 wherein the second core features or elements are positioned equidistantly with respect to a center of the fiber.","120.The optical fiber according to claim 119 wherein the second core features are of similar size.","121.The optical fiber according to claim 117 wherein the size of the second core features is larger or equal to the size of the first cladding features.","122.The optical fiber according to claim 117 wherein the size of the second core features is smaller or larger than the size of the first cladding features.","123.The optical fiber according to claim 104 wherein the optical fiber comprises silica glass and Nco,2 is equal to or larger than 1.444.124.The optical fiber according to claim 104 wherein the number of first cladding features is in the range from 6 to 18.125.The optical fiber according to claim 106 wherein the number of second core features is in the range from 6 to 18.126.The optical fiber according to claim 125 wherein the second core features are characterized by a representative center-to-center distance, Λco,2 between two adjacent second core features.","127.The optical fiber according to claim 126 wherein the second core features are characterized by a cross-sectional dimension in the range from 0.2Λco,2 to 0.9Λco,2.128.The optical fiber according to claim 104 wherein Nco,1 is in the range from 1.45 to 1.49, and Ncl,1 is in the range from 1.43 to 1.45, and Nco,2 is in the range from 1.44 to 1.47, and Ncl,2 is in the range from 1.43 to 1.45.129.The optical fiber according to claim 104 wherein the first core region is micro-structured having one or more spaced apart first core features or elements that are elongated in the fiber axial direction and disposed in the first core material, each said first core feature having a refractive index being lower than Nco,1, whereby a resultant geometrical index Nge,co,1 of the first core region is lowered compared to Nco,1, and wherein Nge,co,1>Nge,cl,1.130.The optical fiber according to any one of the claim 104 wherein the first core region is micro-structured having one or more spaced apart first core features or elements that are elongated in the fiber axial direction and disposed in the first core material, each said first core feature or element having a refractive index being higher than Nco,1, whereby a resultant geometrical index Nge,co,1 of the first core region is higher compared to Nco,1.131.The optical fiber according to claim 130 wherein the optical fiber comprises one first core feature or element.","132.The optical fiber according to claim 131 wherein the first core feature or element is not making contact with the first cladding features or elements.","133.The optical fiber according to claim 131 wherein the first core feature or element has a diameter in the range from 2 μm to 4 μm, such as in the range from 2.5 μm to 3.0 μm.","134.The optical fiber according to claim 131 wherein the first core feature or element has a diameter in the range from 0.5 to 0.8 times a center-to-center distance between two adjacent first cladding features or elements.","135.The optical fiber according to claim 131 wherein the first core region has a background material being identical to the first cladding background material.","136.The optical fiber according to claim 129 wherein Nge,co,1 is larger than Nge,co,2.137.The optical fiber according to claim 129 wherein Nge,co,1 is identical to Nge,co,2.138.The optical fiber according to claim 129 wherein Nge,co,1 is lower than Nge,co,2.139.The optical fiber according to claim 104 wherein the first core material is a background material of the first core region, the first cladding material is a background material of the first cladding region, the second core material is a background material of the second core region, and/or the second cladding material is a background material of the second cladding region.","140.The optical fiber according to claim 104 wherein the first cladding region has a substantially annular shape in the cross section.","141.The optical fiber according to claim 104 wherein the second core region has a substantially annular shape in the cross section.","142.The optical fiber according to claim 104 wherein the second cladding region has a substantially annular shape in the cross section.","143.The optical fiber according to claim 104 wherein the fiber is dimensioned for transmitting or guiding light of one or more predetermined wavelengths.","144.The optical fiber according to claim 143 wherein one of said predetermined wavelengths is in the range from 1.3 μm to 1.7 μm, such as in the range of 1.5 μm to 1.62 μm.","145.The optical fiber according to claim 104 wherein said first core region has a first inscribed core diameter, Dco,1, being larger than 3 times said predetermined wavelength, such as larger than 5 times said predetermined wavelength, such as larger than 7 times said predetermined wavelength, such as larger than 10 times said predetermined wavelength.","146.The optical fiber according to claim 104 wherein said first core region has a first inscribed core diameter, Dco,1, in the range from 4 μm to 25 μm, such as in the range from 4.0 μm to 5.0 μm, such as in the range from 5.0 μm to 6.5 μm, such as in the range from 6.5 μm to 10.0 μm, such as in the range from 10.0 μm to 25.0 μm.","147.The optical fiber according to claim 104 wherein said first core region has a varying refractive index profile, said varying refractive index profile having a highest refractive index equal to Nco,1 and said varying index profile being characterized by an α-profile, where α is in the range from 0 to 100, such as α equal to 2, 3 or higher.","148.The optical fiber according to claim 106 wherein said second core features are positioned substantially circularly symmetric with respect to a center of said first core region.","149.The optical fiber according to claim 106 wherein said second core features are arranged in a single layer surrounding said first core region, such that a distance from a second core feature to a center of said first core region is substantially identical for all second core features.","150.The optical fiber according to claim 106 wherein said second core features are arranged in two or more layers surrounding said first core region.","151.An optical fiber according to claim 106 wherein Λco,2 is in the range from 0.2 μm to 20 μm.","152.The optical fiber according to claim 104 wherein said first cladding features are positioned substantially circularly symmetric with respect to a center of said first core region.","153.The optical fiber according to claim 104 wherein said first cladding features are arranged in a single layer surrounding said first core region, such that a distance from a first cladding feature to a center of said first core region is substantially identical for all first cladding features.","154.The optical fiber according to claim 104 wherein said first cladding features are arranged in two or more layers surrounding said first core region.","155.The optical fiber according to claim 104 wherein the number of said first cladding features is equal to or larger than 3, such as equal to or larger than 6, such as equal to or larger than 8, such as equal to or larger than 18.156.The optical fiber according to claim 104 wherein said first cladding features have a diameter Dcl,1 and a center-to-center spacing between nearest first cladding features of Λcl,1, and Dcl,1/Λcl,1 is in the range from 0.2 to 0.8, such as from 0.4 to 0.6.157.The optical fiber according to claim 156 wherein Dcl,1 is in the range from 0.1 μm to 10 μm.","158.The optical fiber according to claim 156 wherein Λcl,1 is in the range from 0.2 μm to 20 μm, such as from 3.0 μm to 5.0 μm.","159.The optical fiber according to claim 106 wherein said second cladding features have a diameter Dcl,2 and a center-to-center spacing between nearest second cladding features of Λcl,2, where Dcl,2/Λcl,2 is in the range from 0.01 to 0.5, such as from 0.1 to 0.2.160.The optical fiber according to claim 159 wherein Dcl,2 is in the range from 0.1 μm to 5 μm.","161.The optical fiber according to claim 159 wherein Λcl,2 is in the range from 0.2 μm to 20 μm.","162.The optical fiber according to claim 159 wherein Λcl,1 is larger than Λcl,2, such as larger than or equal to 3Λcl,2.163.The optical fiber according to claim 159 wherein Λcl,1 is substantially identical to Λcl,2.164.The optical fiber according to claim 159 wherein Dcl,1 is larger than Dcl,2.165.The optical fiber according to claim 159 wherein Dcl,1/Λcl,1 and Dcl,2/Λcl,2 are substantially identical.","166.The optical fiber according to claim 103 wherein said optical fiber comprises one or more glass materials.","167.The optical fiber according to claim 166 wherein said optical fiber comprises silica.","168.The optical fiber according to claim 103 wherein said optical fiber comprises polymer.","169.The optical fiber according to claim 166 wherein said first cladding features, second core features, and/or second cladding features are voids comprising vacuum, air, or another gas.","170.The optical fiber according to claim 104 wherein said optical fiber has a non-circular shape of the outer cladding, such as a non-polygonal shape, such as an elliptical shape.","171.The optical fiber according to claim 170 wherein said outer shape has a predetermined orientation in the cross section, the predetermined orientation being determined from the position of said first cladding features.","172.The optical fiber according to claim 104 wherein said first core region has a non-circular shape in the cross section, such as an elliptical shape, providing a high birefringence in the optical fiber.","173.The optical fiber according to claim 104 wherein said first cladding region has a non-circular, inner shape in the cross section, such as an elliptical, inner shape, providing a high birefringence in the optical fiber.","174.The optical fiber according to claim 104 wherein said optical fiber guides light at a predetermined wavelength in a single mode.","175.The optical fiber according to claim 104 wherein said optical fiber guides light at a predetermined wavelength in a higher order mode.","176.The optical fiber according to claim 104 wherein the second core has a diameter larger than 10 μm, such as in the range from 10 μm to 15 μm, such as in the range from 15 μm to 20 μm.","177.The optical fiber according to claim 104 wherein the first core has a diameter larger than 3 μm, such as in the range from 4 μm to 8 μm, such as in the range from 4 μm to 6 μm.","178.The optical fiber according to claim 104 wherein the first core and the second core are separated by a distance of at least 3 μm, such as at least 4 μm.","179.The optical fiber according to claim 104 wherein the first core has a non-zero distance to a low-index feature of the first cladding, such as a distance larger than 0.5 μm, such as larger than 1.0 μm.","180.The optical fiber according to claim 104 wherein light is propagating substantially in the first core.","181.The optical fiber according to claim 104 wherein light is coupled substantially to the first core.","182.The optical fiber according to claim 104 wherein the second core has a larger area than the first core.","183.The optical fiber according to claim 104 wherein the second core has an effective refractive index, nco2,eff being larger than (nco,1,eff+ncl,2,eff)/2 or larger than (nco,1,eff+ncl,1,eff)/2, where nco,1,eff is an effective refractive index of the first core region, ncl,1,eff is an effective refractive index of the first cladding region, and ncl,2,eff is an effective refractive index of the second cladding region.","184.The optical fiber according to claim 104 wherein the second core has a mean or geometric refractive index, nco2,g being larger than (nco,1,g+ncl,2,g)/2 or larger than (nco,1,g+ncl,1,g)/2, where nco,1,g is a mean or geometric refractive index of the first core region, ncl,1,g is a mean or geometric refractive index of the first cladding region, and ncl,2,g is a mean or geometric refractive index of the second cladding region.","185.The optical fiber according to claim 104 wherein said fiber is dimensioned to guide light in a single mode in the first core region at a first wavelength, λ1, being in the range from 1500 to 1640 nm, and said optical fiber exhibits negative dispersion at λ1.186.The optical fiber according to claim 185 wherein the optical fiber is dimensioned to guide light in the second core region for light shorter than a cut-off wavelength, λc, where λc is more than 30 nm shorter than λ1.187.An article comprising a module, said module being employed for dispersion compensation in an optical communication system, said module comprising an optical fiber according to claim 104.188.An optical fiber for transmitting light, said optical fiber having an axial direction and a cross section perpendicular to said axial direction, said optical fiber comprising: (1) a first core region comprising a first core material having a refractive index Nco,1 and at least one first core feature having a refractive being lower than Nco,1, whereby a resultant geometrical index Nge,co,1 of the first core region is lowered compared to Nco,1, (2) a micro-structured first cladding region, said first cladding region surrounding the first core region, and said first cladding region comprising a first cladding material and a plurality of spaced apart first cladding features that are elongated in the fiber axial direction and disposed in the first cladding material, said first cladding material having a refractive index Ncl,1 and each said first cladding feature having a refractive index being lower than Ncl,1, whereby a resultant geometrical index Nge,cl,1 of the first cladding region is lowered compared to Ncl,1, (3) a second core region, said second core region surrounding said first cladding region, and said second core region comprising a second core material having a refractive index Nco,2, and (4) a micro-structured second cladding region, said second cladding region surrounding the second core region, and said second cladding region comprising a second cladding material and a plurality of spaced apart second cladding features that are elongated in the fiber axial direction and disposed in the second cladding material, said second cladding material having a refractive index Ncl,2 and each said second cladding feature having an refractive index being lower than Ncl,2, whereby a resultant geometrical index Nge,cl,2 of the second cladding region is lowered compared to Ncl,2, wherein the first core material, the first cladding material and the first cladding features, the second core material, and the second cladding material and the second cladding features are selected and arranged so that Nco,1>Nge,cl,1, Nco,2>Nge,cl,1, Nco,2>Nge,cl,2, Nge,co,1<Nge,cl,2, and the second cladding features are placed in a substantially two dimensionally periodic structure, whereby the second cladding region exhibits photonic band gap effect.","189.The optical fiber according to claim 188 wherein said second cladding features are placed in a substantially two dimensional periodic structure.","190.The optical fiber according to claim 188 wherein said second cladding features are placed in a radial periodic structure.","191.The optical fiber according to claim 188 wherein the second core has a diameter larger than 10 μm, such as in the range from 10 μm to 15 μm, such as in the range from 15 μm to 20 μm.","192.The optical fiber according to claim 188 wherein the first core has a diameter larger than 3 μm, such as in the range from 4 μm to 8 μm, such as in the range from 4 μm to 6 μm.","193.The optical fiber according to claim 188 wherein the first core and the second core are separated by a distance of at least 3 μm, such as at least 4 μm.","194.The optical fiber according to claim 188 wherein the first core has a non-zero distance to a low-index feature of the first cladding, such as a distance larger than 0.5 μm, such as larger than 1.0 μm.","195.The optical fiber according to claim 188 wherein light is propagating substantially in the first core.","196.The optical fiber according to claim 188 wherein light is coupled substantially to the first core.","197.The optical fiber according to claim 188 wherein the second core has a larger area than the first core.","198.The optical fiber according claim 188 wherein the second core has an effective refractive index, nco2,eff being larger than (nco,1,eff+ncl,2,eff)/2 or larger than (nco,1,eff+ncl,1,eff)/2, where nco,1,eff is an effective refractive index of the first core region, ncl,1,eff is an effective refractive index of the first cladding region, and ncl,2,eff is an effective refractive index of the second cladding region.","199.The optical fiber according to claim 188 wherein the second core has a mean or geometric refractive index, nco2,g being larger than (nco,1,g+ncl,2,g)/2 or larger than (nco,1,g+ncl,1,g)/2, where nco,1,g is a mean or geometric refractive index of the first core region, ncl,1,g is a mean or geometric refractive index of the first cladding region, and ncl,2,g is a mean or geometric refractive index of the second cladding region.","200.The optical fiber according to claim 188 wherein said fiber is dimensioned to guide light in a single mode in the first core region at a first wavelength, λ1, being in the range from 1500 to 1640 nm, and said optical fiber exhibits negative dispersion at λ1.201.The optical fiber according to claim 200 wherein the optical fiber is dimensioned to guide light in the second core region for light shorter than a cut-off wavelength, λc, where λc is more than 30 nm shorter than λ1.202.An article comprising a module, said module being employed for dispersion compensation in an optical communication system, said module comprising an optical fiber according to claim 188.203.An optical transmission line, comprising at least: (1) a length of standard transmission fiber with a positive dispersion and a positive dispersion slope at least at a wavelength, λ1, of 1550 nm, and (2) a length of dispersion compensating fiber, such as a length of dispersion compensating fiber spooled in a fiber module, wherein said dispersion compensating has negative dispersion with a numerical value of the dispersion being larger than 500 ps/nm/km at λ1.204.An optical transmission line, comprising at least: (1) a length of standard transmission fibre with a positive dispersion and a positive dispersion slope at least at a wavelength, λ1, of 1550 nm, and (2) a length of dispersion compensating fibre, such as a length of dispersion compensating fibre according to claim 188 spooled in a fibre module, wherein said dispersion compensating has negative dispersion with a numerical value of the dispersion being larger than 500 ps/nm/km at λ1.205.The optical transmission line according to claim 203 wherein the dispersion compensating fiber has a relative dispersion slope of more than 0.01 nm−1, such as more than 0.02 nm−1.206.An optical fiber for transmitting or guiding light, said optical fiber having an axial direction and a cross section perpendicular to said axial direction, said optical fiber comprising: (1) a first core region comprising a first core material having a refractive index Nco,1, (2) a microstructured first cladding region surrounding the first core region, said first cladding region comprising a first cladding material and a plurality of spaced apart first cladding features or elements that are elongated in the fiber axial direction and disposed in the first cladding material, said first cladding material having a refractive index Ncl,1 and each said first cladding feature or element having a refractive index being lower than Ncl,1, whereby a resultant geometrical index Nge,cl,1 of the first cladding region is lowered compared to Ncl,1, (3) a second core region surrounding said first cladding region, said second core region comprising a second core material having a refractive index Nco,2, and (4) a micro-structured second cladding region surrounding the second core region, said second cladding region comprising a second cladding material and a plurality of spaced apart second cladding features or elements that are elongated in the fiber axial direction and disposed in the second cladding material, said second cladding material having a refractive index Ncl,2 and each said second cladding feature or element having an refractive index being lower than Ncl,2, whereby a resultant geometrical index Nge,cl,2 of the second cladding region is lowered compared to Ncl,2, wherein the first core material, the first cladding material and the first cladding features or elements, the second core material, and the second cladding material and the second cladding features or elements are selected and arranged so that Ncl,1>Nge,cl,1, Nco,2>Nge,cl,1, and Nco,2>Nge,cl,2.207.The optical fiber according to claim 206 wherein the second core region is micro-structured having a plurality of spaced apart second core features or elements that are elongated in the fiber axial direction and disposed in the second core material, each said second core feature or element having a refractive index being higher than Nco,2, whereby a resultant geometrical index Nge,co,2 of the second core region is higher compared to Nco,2.208.The optical fiber according to claim 207 wherein Nco,1 is larger than Nge,co,2.209.The optical fiber according to claim 207 wherein Nco,1 is identical to Nge,co,2.210.The optical fiber according to claim 207 wherein Nco,1 is lower than Nge,co,2.211.The optical fiber according to claim 207 wherein the number of said second core features is equal to or larger than 6, such as equal to or larger than 18.212.The optical fiber according to claim 207 wherein said second core features have a diameter Dco,2 and a center-to-center spacing between nearest second core features of Λco,2, and Dco,2/Λco,2 is in the range from 0.01 to 0.5, such as from 0.1 to 0.2.213.The optical fiber according to claim 212 wherein Dco,2 is in the range from 0.1 μm to 5 μm.","214.The optical fiber according to claim 206 wherein said first cladding features have a diameter Dcl,1 and a center-to-center spacing between nearest first cladding features of Λcl,1, and Dcl,1/Λcl,1 is in the range from 0.2 to 0.8, such as from 0.4 to 0.6.215.The optical fiber according to claim 214 wherein Dcl,1/Λcl,1 is larger than Dco,2/Λco,2.216.An optical fiber for transmitting light, said optical fiber having an axial direction and a cross section perpendicular to said axial direction, said optical fiber comprising: (1) a first core region comprising a first core material having a refractive index Nco,1 and at least one first core feature having a refractive being lower than Nco,1, whereby a resultant geometrical index Nge,co,1 of the first core region is lowered compared to Nco,1, (2) a micro-structured first cladding region, said first cladding region surrounding the first core region, and said first cladding region comprising a first cladding material and a plurality of spaced apart first cladding features that are elongated in the fiber axial direction and disposed in the first cladding material, said first cladding material having a refractive index Ncl,1 and each said first cladding feature having a refractive index being lower than Ncl,1, whereby a resultant geometrical index Nge,cl,1 of the first cladding region is lowered compared to Ncl,1, (3) a second core region, said second core region surrounding said first cladding region, and said second core region comprising a second core material having a refractive index Nco,2, and (4) a micro-structured second cladding region, said second cladding region surrounding the second core region, and said second cladding region comprising a second cladding material and a plurality of spaced apart second cladding features that are elongated in the fiber axial direction and disposed in the second cladding material, said second cladding material having a refractive index Ncl,2 and each said second cladding feature having an refractive index being lower than Ncl,2, whereby a resultant geometrical index Nge,cl,2 of the second cladding region is lowered compared to Ncl,2, wherein the first core material, the first cladding material and the first cladding features, the second core material, and the second cladding material and the second cladding features are selected and arranged so that Nco,1>Nge,cl,1, Nco,2>Nge,cl,1, Nco,2>Nge,cl,2, Nge,co,1<Nge,cl,2, and the second cladding features are placed so that the second cladding region exhibits photonic band gap effect.","217.The optical fiber according to claim 216 wherein the second core has a diameter larger than 10 μm, such as in the range from 10 μm to 15 μm, such as in the range from 15 μm to 20 μm.","218.The optical fiber according to claim 216 wherein the first core has a diameter larger than 3 μm, such as in the range from 4 μm to 8 μm, such as in the range from 4 μm to 6 μm.","219.The optical fiber according to claim 216 wherein the first core and the second core are separated by a distance of at least 3 μm, such as at least 4 μm.","220.The optical fiber according to claim 216 wherein the first core has a non-zero distance to a low-index feature of the first cladding, such as a distance larger than 0.5 μm, such as larger than 1.0 μm.","221.The optical fiber according to claim 216 wherein light is propagating substantially in the first core.","222.The optical fiber according to claim 216 wherein light is coupled substantially to the first core.","223.The optical fiber according to claim 216 wherein the second core has a larger area than the first core.","224.The optical fiber according to claim 216 wherein the second core has an effective refractive index, nco2,eff being larger than (nco,1,eff+ncl,2,eff)/2 or larger than (nco,1,eff+ncl,1,eff)/2, where nco,1,eff is an effective refractive index of the first core region, ncl,1,eff is an effective refractive index of the first cladding region, and ncl,2,eff is an effective refractive index of the second cladding region.","225.The optical fiber according to claim 216 wherein the second core has a mean or geometric refractive index, nco2,g being larger than (nco,1,g+ncl,2,g)/2 or larger than (nco,1,g+ncl,1,g)/2, where nco,1,g is a mean or geometric refractive index of the first core region, ncl,1,g is a mean or geometric refractive index of the first cladding region, and ncl,2,g is a mean or geometric refractive index of the second cladding region.","226.The optical fiber according to claim 216 wherein said fiber is dimensioned to guide light in a single mode in the first core region at a first wavelength, λ1, being in the range from 1500 to 1640 nm, and said optical fiber exhibits negative dispersion at λ1.227.The optical fiber according to claim 226 wherein the optical fiber is dimensioned to guide light in the second core region for light shorter than a cut-off wavelength, λc, where λc is more than 30 nm shorter than λ1.228.An article comprising a module, said module being employed for dispersion compensation in an optical communication system, said module comprising an optical fiber according to claim 216.229.The article according to claim 228 wherein the dispersion compensating fiber has a relative dispersion slope of more than 0.01 nm−1, such as more than 0.02 nm−1.230.An optical transmission line, comprising at least: (1) a length of standard transmission fiber with a positive dispersion and a positive dispersion slope at least at a wavelength, λ1, of 1550 nm, and (2) a length of dispersion compensating fiber, such as a length of dispersion compensating fiber spooled in a fiber module, wherein said dispersion compensating has negative dispersion with a numerical value of the dispersion being larger than 500 ps/nm/km at λ1.231.An optical transmission line, comprising at least: (1) a length of standard transmission fibre with a positive dispersion and a positive dispersion slope at least at a wavelength, λ1, of 1550 nm, and (2) a length of dispersion compensating fibre, such as a length of dispersion compensating fibre according to claim 216 spooled in a fibre module, wherein said dispersion compensating has negative dispersion with a numerical value of the dispersion being larger than 500 ps/nm/km at λ1.232.The optical transmission line according to claim 230 wherein the dispersion compensating fiber has a relative dispersion slope of more than 0.01 nm−1, such as more than 0.02 nm−1.233.An article comprising a module, said module being employed for dispersion compensation in an optical communication system, said module comprising an optical fiber according to claim 216.234.An optical fiber having an axial direction and a cross section perpendicular to said axial direction, the optical fiber having negative dispersion in the vicinity of a predetermined wavelength and being employed for dispersion compensation; CHARACTERIZED IN THAT; said optical fiber comprises: (1) a first core region being positioned substantially in a center of the optical fiber, the first core region comprising a first core material of refractive index, Nco,1; 2) a first cladding region surrounding the first core region, the first cladding region comprises a multiplicity of spaced apart first cladding features that are elongated in the axial direction and disposed in the first cladding material, said first cladding material having a refractive index Ncl,1; (3) a second core region surrounding the first cladding region, the second core region comprising a second core material of refractive index, Nco,2, and the second core region having a substantially annular shape in the cross section; (4) a second cladding region surrounding the second core region, the second cladding region comprising a second cladding material of refractive index, Ncl,2, and the second cladding region having a substantially annular shape in the cross section.","235.The optical fiber according to claim 234 wherein said predetermined wavelength is in a range from 1.3 μm to 1.7 μm, such as from around 1.5 μm to 1.62 μm.","236.The optical fiber according to claim 235 wherein Nco,1 is larger than Nco,2.237.The optical fiber according to claim 235 wherein Nco,1 is identical to Nco,2.238.The optical fiber according to claim 235 wherein Nco,1 is lower than Nco,2.239.The optical fiber according to claim 234 wherein said first cladding region has a substantially annular shape in the cross section.","240.The optical fiber according to claim 234 wherein said optical fiber has a first inscribed core diameter, Dco,1, being larger than 3 times said predetermined wavelength, such as larger than 5 times said predetermined wavelength, such as larger than 7 times said predetermined wavelength, such as larger than 10 times said predetermined wavelength.","241.The optical fiber according to claim 234 wherein Dco,1 is in the range from 4 μm to 25 μm, such as in the range from 4.0 μm to 5.0 μm, such as in the range from 5.0 μm to 6.5 μm, such as in the range from 6.5 μm to 10.0 μm, such as in the range from 10.0 μm to 25.0 μm.","242.The optical fiber according to claim 241 wherein said first core region has a varying refractive index profile, said varying refractive index profile having a highest refractive index equal to Nco,1, and said varying index profile being characterized by an α-profile, where α is in the range from 0 to 100, such as α equal to 2, 3 or higher.","243.The optical fiber according to claim 234 wherein said second core region is a homogeneous region.","244.The optical fiber according to claim 234, wherein said second core region comprises a multiplicity of spaced apart second core features that are elongated in the axial direction and disposed in the second core material.","245.The optical fiber according to claim 244 wherein said second core features are positioned substantially circularly symmetric with respect to a center of said first core region.","246.The optical fiber according to claim 234 wherein said second core features are arranged in a single layer surrounding said first core region, such that a distance from a second core feature to a center of said first core region is substantially identical for all second core features.","247.The optical fiber according to claim 244 wherein said second core features are arranged in two or more layers surrounding said first core region.","248.The optical fiber according to claim 244 wherein the number of said second core features is equal to or larger than 6, such as equal to or larger than 18.249.The optical fiber according to claim 244 wherein said second core features have a diameter Dco,2 and a center-to-center spacing between nearest second core features of Λco,2, and Dco,2/Λco,2 is in the range from 0.01 to 0.5, such as from 0.1 to 0.2.250.The optical fiber according to claim 249 wherein Dco,2 is in the range from 0.1 μm to 5 μm.","251.The optical fiber according to claim 249 wherein Λco,2 is in the range from 0.2 μm to 20 μm.","252.The optical fiber according to claim 234 wherein said first cladding features are positioned substantially circularly symmetric with respect to a center of said first core region.","253.The optical fiber according to claim 234 wherein said first cladding features are arranged in a single layer surrounding said first core region, such that a distance from a first cladding feature to a center of said first core region is substantially identical for all first cladding features.","254.The optical fiber according to claim 234 wherein said first cladding features are arranged in two or more layers surrounding said first core region.","255.The optical fiber according to claim 234 wherein the number of said first cladding features is equal to or larger than 3, such as equal to or larger than 6, such as equal to or larger than 8, such as equal to or larger than 18.256.The optical fiber according to claim 234 wherein said first cladding features have a diameter Dcl,1 and a center-to-center spacing between nearest first cladding features of Λcl,1, and Dcl,1/Λcl,1 is in the range from 0.2 to 0.8, such as from 0.4 to 0.6.257.The optical fiber according to claim 256, wherein Dcl,1 is in the range from 0.1 μm to 10 μm.","258.The optical fiber according to claim 256 wherein Λcl,1 is in the range from 0.2 μm to 20 μm.","259.The optical fiber according to claim 256 wherein Dcl,1/Λcl,1 is larger than Dco,2/Λco,2.260.The optical fiber according to claim 234 wherein said second cladding region comprises a multiplicity of spaced apart second cladding features that are elongated in the axial direction and disposed in said second cladding material.","261.The optical fiber according to claim 260 wherein said second cladding features have a diameter Dcl,2 and a center-to-center spacing between nearest second cladding features of Λcl,2, where Dcl,2/Λcl,2 is in the range from 0.01 to 0.5, such as from 0.1 to 0.2.262.The optical fiber according to claim 261 wherein Dcl,2 is in the range from 0.1 μm to 5 μm.","263.The optical fiber according to claim 261 wherein Λcl,2 is in the range from 0.2 μm to 20 μm.","264.The optical fiber according to claim 261 wherein Λcl,1 is larger than Λcl,2, such as larger than or equal to 3Λcl,2.265.The optical fiber according to claim 261 wherein Λcl,1 is substantially identical to Λcl,2.266.The optical fiber according to claim 261 wherein Dcl,1 is larger than Dcl,2.267.The optical fiber according to claim 261 wherein Dcl,1/Λcl,1 and Dcl,2/Λcl,2 are substantially identical.","268.The optical fiber according to claim 234 wherein said optical fiber comprises one or more glass materials.","269.The optical fiber according to claim 268 wherein said optical fiber comprises silica.","270.The optical fiber according to claim 234 wherein said optical fiber comprises polymer.","271.The optical fiber according to claim 268 wherein said first cladding features, second core features, and/or second cladding features are voids comprising vacuum, air, or another gas.","272.The optical fiber according to claim 234 wherein said optical fiber has a non-circular shape of the outer cladding, such as a non-polygonal shape, such as an elliptical shape.","273.The optical fiber according to claim 272 wherein said outer shape has a predetermined orientation in the cross section, the predetermined orientation being determined from the position of said first cladding features.","274.The optical fiber according to claim 234 wherein said first core region has a non-circular shape in the cross section, such as an elliptical shape, providing a high birefringence in the optical fiber.","275.The optical fiber according to claim 234 wherein said first cladding region has a non-circular, inner shape in the cross section, such as an elliptical, inner shape, providing a high birefringence in the optical fiber.","276.The optical fiber according to claim 234 wherein said optical fiber guides light at predetermined wavelength in a single mode.","277.The optical fiber according to claim 234 wherein said optical fiber guides light at predetermined wavelength in a higher order mode.","278.The optical fiber according to claim 234 wherein the second core has a diameter larger than 10 μm, such as in the range from 10 μm to 15 μm, such as in the range from 15 μm to 20 μm.","279.The optical fiber according to claim 234 wherein the first core has a diameter larger than 3 μm, such as in the range from 4 μm to 8 μm, such as in the range from 4 μm to 6 μm.","280.The optical fiber according to claim 234 wherein the first core and the second core are separated by a distance of at least 3 μm, such as at least 4 μm.","281.The optical fiber according to claim 234 wherein the first core has a non-zero distance to a low-index feature of the first cladding, such as a distance larger than 0.5 μm, such as larger than 1.0 μm.","282.The optical fiber according to claim 234 wherein light is propagating substantially in the first core.","283.The optical fiber according to claim 234 wherein light is coupled substantially to the first core.","284.The optical fiber according to claim 234 wherein the second core has a larger area than the first core.","285.The optical fiber according to claim 234 wherein the second core has an effective refractive index, nco2,eff being larger than (nco,1,eff+ncl,2,eff)/2 or larger than (nco,1,eff+ncl,1,eff)/2, where nco,1,eff is an effective refractive index of the first core region, ncl,1,eff is an effective refractive index of the first cladding region, and ncl,2,eff is an effective refractive index of the second cladding region.","286.The optical fiber according to claim 234 wherein the second core has a mean or geometric refractive index, nco2,g being larger than (nco,1,g+ncl,2,g)/2 or larger than (nco,1,g+ncl,1,g)/2, where nco,1,g is a mean or geometric refractive index of the first core region, ncl,1,g is a mean or geometric refractive index of the first cladding region, and ncl,2,g is a mean or geometric refractive index of the second cladding region.","287.The optical fiber according to claim 234 wherein said fiber is dimensioned to guide light in a single mode in the first core region at a first wavelength, λ1, being in the range from 1500 to 1640 nm, and said optical fiber exhibits negative dispersion at λ1.288.The optical fiber according to claim 287 wherein the optical fiber is dimensioned to guide light in the second core region for light shorter than a cut-off wavelength, λc, where λc is more than 30 nm shorter than λ1.289.An article comprising a module, said module being employed for dispersion compensation in an optical communication system, said module comprising an optical fiber according to claim 234.290.An optical transmission line, comprising at least: (1) a length of standard transmission fiber with a positive dispersion and a positive dispersion slope at least at a wavelength, λ1, of 1550 nm, and (2) a length of dispersion compensating fiber, such as a length of dispersion compensating fiber spooled in a fiber module, wherein said dispersion compensating has negative dispersion with a numerical value of the dispersion being larger than 500 ps/nm/km at λ1.291.An optical transmission line, comprising at least: (1) a length of standard transmission fibre with a positive dispersion and a positive dispersion slope at least at a wavelength, λ1, of 1550 nm, and (2) a length of dispersion compensating fibre, such as a length of dispersion compensating fibre according to claim 234 spooled in a fibre module, wherein said dispersion compensating has negative dispersion with a numerical value of the dispersion being larger than 500 ps/nm/km at λ1.292.The optical transmission line according to claim 290 wherein the dispersion compensating fiber has a relative dispersion slope of more than 0.01 nm−1, such as more than 0.02 nm−1 claim 293.An article comprising a module, said module being employed for dispersion compensation in an optical communication system, said module comprising an optical fiber according to claim 234."],["<SOH> 1.BACKGROUND OF THE INVENTION <EOH>The present invention relates to new designs of microstructured fibers providing improvements with respect to dispersion compensation (including dispersion slope compensation) for fiber optical communication systems and with respect to non-linear optical fibers for applications at near-infrared wavelengths."],["<SOH> SUMMARY OF THE INVENTION <EOH>In a first aspect, the present invention relates to an optical fiber having an axial direction and a cross section perpendicular to said axial direction.","The optical fiber has negative dispersion in the vicinity of a predetermined wavelength.","The optical fiber is characterised by a number of regions: 1) a first core region being positioned substantially in a center of the optical fiber, the first core region comprising a first core material of refractive index, N co,1 .","In the present context, “positioned substantially” means that it is desired to have the first core placed in the center, but manners of production may introduce minor structural deviations such that a center of the first core region and a center of the whole optical fiber may not coincide.","Also, the optical fiber may have a non-circular outer shape in the cross section, making the definition of a center of the optical fiber less stringent.","In the case of a non-circular outer shape, the center is determined as the center-of-gravity of a two-dimensional element with the same outer shape as the optical fiber.","2) a first cladding region surrounding the first core region, the first cladding region comprises a multiplicity of spaced apart first cladding features that are elongated in the axial direction and disposed in the first cladding material, said first cladding material having a refractive index N cl,1 ; 3) a second core region surrounding the first cladding region, the second core region comprising a second core material of refractive index, N co,2 , and the second core region having a substantially annular shape in the cross section.","Since the interface between the inner cladding and the outer core may be an interface where at least on one side there is a microstructure (the inner cladding region), the interface may not necessarily be seen as a smooth interface (e.g.","in the case of the two regions having identical background refractive indices).","Therefore, the shape of the outer core region may not necessarily be a viewed as a smooth ring-like shape—hence the wording “substantially annular shape”; 4) a second cladding region surrounding the second core region, the second cladding region comprising a second cladding material of refractive index, N cl,2 , and the second cladding region having a substantially annular shape in the cross section.","In the present context, the wording “substantially annular shape” is used for the same reasons as discussed above.","Typically, the predetermined wavelength is in a range from 1.3 μm to 1.7 μm, such as from around 1.5 μm to 1.62 μm.","In a preferred embodiment, the optical fiber has a higher refractive index in the first core region than in the second core region.","In another preferred embodiment, the optical fiber has identical refractive index in the first core region and in the second core region.","This allows for example to manufacture the fiber in a single material, such as e.g.","pure silica.","In a preferred embodiment, the optical fiber has a lower refractive index in the first core region than in the second core region.","This allows for example to have a very large mode field diameter.","In another preferred embodiment, the first cladding region has a substantially annular shape in the cross section.","The wording “substantially” being used for the same reasons as previously discussed.","In another preferred embodiment, the optical fiber has a first inscribed core diameter, D co,1 , being larger than 3 times said predetermined wavelength, such as larger than 5 times said predetermined wavelength, such as larger than 7 times said predetermined wavelength, such as larger than 10 times said predetermined wavelength.","In another preferred embodiment, D co,1 is in the range from 4 μm to 25 μm, such as in the range from 4.0 μm to 5.0 μm, such as in the range from 5.0 μm to 6.5 μm, such as in the range from 6.5 μm to 10.0 μm, such as in the range from 10.0 μm to 25.0 μm.","In another preferred embodiment, the first core region has a varying refractive index profile, said varying refractive index profile having a highest refractive index equal to N co,1 , and said varying index profile being characterised by an □-profile, where □ is in the range from 0 to 100, such as □ equal to 2, 3 or higher.","In another preferred embodiment, the second core region is a homogeneous region.","In another preferred embodiment, the second core region is microstructured.","This increases the effective refractive index ranges that are available for the second core region without the use of doping.","In another preferred embodiment, the microstructured features in the second core are positioned substantially circularly symmetric with respect to a center of said first core region.","This is preferred in order to lower polarisation mode dispersion (PMD) in the fiber.","An optical fiber according to any one of the preceding claims, wherein said second core features are arranged in a single layer surrounding said first core region, such that a distance from a second core feature to a center of said first core region is substantially identical for all second core features.","This is preferred in order to lower PMD, and further preferred for ease of fabrication.","In another preferred embodiment, the second core features are arranged in two or more layers surrounding said first core region.","This is preferred to reduce bending losses.","In another preferred embodiment, the number of second core features is equal to or larger than 6, such as equal to or larger than 18.A high number is preferred to modify the effective refractive index of the second core region.","In another preferred embodiment, the second core features have a diameter D co,2 and a center-to-center spacing between nearest second core features of Λ co,2 , and D co,2 /Λ co,2 is in the range from 0.01 to 0.5, such as from 0.1 to 0.2.Generally, the microstructured features in the second core should be relatively small in order to ensure the possibility of the second core to support a limited number of modes.","In another preferred embodiment, D co,2 is in the range from 0.1 μm to 5 μm.","Typically, this is the design parameter range that provides strongest dispersion.","In another preferred embodiment, λ/Λ co,2 is in the range from 0.2 μm to 20 μm.","Typically, this is the design parameter range that provides strongest dispersion.","In another preferred embodiment, the first cladding features are positioned substantially circularly symmetric with respect to a center of said first core region.","This is preferred to lower PMD in the fiber.","In another preferred embodiment, the first cladding features are arranged in a single layer surrounding said first core region, such that a distance from a first cladding feature to a center of said first core region is substantially identical for all first cladding features.","This is preferred to lower PMD in the fiber.","In another preferred embodiment, the said first cladding features are arranged in two or more layers surrounding said first core region.","This is preferred to lower bending losses in the fibers and/or to provide stronger dispersion.","In another preferred embodiment, the number of said first cladding features is equal to or larger than 3, such as equal to or larger than 6, such as equal to or larger than 8, such as equal to or larger than 18.This range of numbers of features allows for tailoring of the birefringence of the fiber to a large degree.","Higher numbers of features may be desired to provide low PMD for example through more circular symmetric feature arrangements.","In another preferred embodiment, the first cladding features have a diameter D cl,1 and a center-to-center spacing between nearest first cladding features of Λ cl,1 , and D cl,1 /Λ cl,1 is in the range from 0.2 to 0.8, such as from 0.4 to 0.6.To provide strong dispersion, relatively large features are required.","Hence, the features should be large enough to provide a buffer region between the two cores.","In another preferred embodiment, D cl,1 is in the range from 0.1 μm to 10 μm.","Typically, this range provides the most attractive design parameter range in order to obtain strong dispersion for wavelengths around 1.55 μm.","In another preferred embodiment, Λ cl,1 is in the range from 0.2 μm to 20 μm.","Typically, this range provides the most attractive design parameter range in order to obtain strong dispersion for wavelengths around 1.55 μm.","In another preferred embodiment, D cl,1 /Λ cl,1 is larger than D co,2 /Λ co,2 .","This is preferred to ensure that the first cladding region acts as a buffer between the two cores.","In another preferred embodiment, the second cladding region comprises a multiplicity of spaced apart second cladding features that are elongated in the axial direction and disposed in said second cladding material.","This provides further means for tailoring the dispersion properties of the fiber.","In another preferred embodiment, second cladding features have a diameter D cl,2 and a center-to-center spacing between nearest second cladding features of □cl,2, where D cl,2 /Λ cl,2 is in the range from 0.01 to 0.5, such as from 0.1 to 0.2.It is often preferred that these features are comparable to the features of the second core.","It may be preferred that the second core is only weakly guiding or has a cut-off close to or around the operational wavelength.","Hence the second cladding features may be preferred to have a limited size in order to not provide strong confinement for the second core—or alternatively not to provide confinement of a high number of modes in the second core.","In another preferred embodiment, D cl,2 is in the range from 0.1 μm to 5 μm.","In accordance with the above-stated preferred embodiment, this is preferred to have the second cladding features being comparable in size to the second core features.","Similarly, it is further preferred that Λ cl,2 is in the range from 0.2 μm to 20 μm.","In another preferred embodiment, Λcl, 1 is larger than Λ cl,2 .","This is preferred in order to allow the first and second cladding regions to have the same background material, but the effective refractive index of the two regions to be different at a given wavelength, such as the predetermined wavelength.","In particular, it is preferred to have Λ cl,1 larger or equal to 3Λ cl,2 as such a ratio may easily be realised using well known methods for fabricating microstructured fibers, such as methods employing stacking of capillary tubes.","In another preferred embodiment, Λ cl,1 is substantially identical to Λ cl,2 .","This may be preferred as similar pitches (i.e.","center-to-center features spacing or lattice period in the case of cladding comprising periodic feature arrangements) often allow simple fabrication, thereby improving reproducibility of the fiber.","In another preferred embodiment, D cl,1 is larger than D cl,2 .","This is preferred in order for the first cladding region to have a lower effective, refractive index than the second cladding.","In another preferred embodiment, D cl,1 /Λ cl,1 and D cl,2 /Λ cl,2 are substantially identical.","This may be preferred as similar hole and pitch sizes provide the simplest fabrication, thereby improving reproducibility of the fiber.","In another preferred embodiment, the optical fiber comprises one or more glass materials.","In another preferred embodiment, the optical fiber comprises silica.","In another preferred embodiment, the optical fiber comprises polymer.","In another preferred embodiment, the first cladding features, second core features, and/or second cladding features are voids comprising vacuum, air, or another gas.","In another preferred embodiment, the optical fiber has a non-circular shape of the outer cladding, such as a non-polygonal shape, such as an elliptical shape.","This may be advantageous for a range of reasons—for example for fibers with high birefringence (for example for handling splicing issues), or to ensure bending in certain preferred directions.","In another preferred embodiment, the outer shape has a predetermined orientation in the cross section, the predetermined orientation being determined from the position of said first cladding features.","This may be preferred to further ensure bending in certain preferred directions with respect to the actual directions in the microstructure in and/or around the core region.","Also for polarisation maintaining, dispersion compensating fibers this is preferred.","In another preferred embodiment, the first core region has a non-circular shape in the cross section, such as an elliptical shape.","This is preferred to provide a high birefringence in the optical fiber.","In another preferred embodiment, the first cladding region has a non-circular, inner shape in the cross section, such as an elliptical, inner shape, providing a high birefringence in the optical fiber.","This is further preferred for polarisation maintaining, dispersion compensating fiber applications.","In another preferred embodiment, the optical fiber guides light at predetermined wavelength in a single mode.","In another preferred embodiment, the optical fiber guides light at predetermined wavelength in a higher order mode.","This is preferred as higher order modes may provide even stronger dispersion than the fundamental mode including the possibility of strong positive dispersion.","In another preferred embodiment, the invention covers an article comprising a module, where the module is employed for dispersion compensation in an optical communication system, and the module comprising an optical fiber according to the invention.","In a second aspect, the present invention relates to an optical fiber having an axial direction and a cross section perpendicular to said axial direction.","The optical fiber has positive dispersion in the vicinity of a predetermined wavelength.","The optical fiber is characterised by a number of regions: 1) a first core region being positioned substantially in a center of the optical fiber, the first core region comprising a first core material of refractive index, N co,1 .","In the present context, “positioned substantially” means that it is desired to have the first core placed in the center, but manners of production may introduce minor structural deviations such that a center of the first core region and a center of the whole optical fiber may not coincide.","Also, the optical fiber may have a non-circular outer shape in the cross section, making the definition of a center of the optical fiber less stringent.","In the case of a non-circular outer shape, the center is determined as the center-of-gravity of a two-dimensional element with the same outer shape as the optical fiber.","2) a first cladding region surrounding the first core region, the first cladding region comprises a multiplicity of spaced apart first cladding features that are elongated in the axial direction and disposed in the first cladding material, said first cladding material having a refractive index N cl,1 ; 3) a second core region surrounding the first cladding region, the second core region comprising a second core material of refractive index, N co,2 , and the second core region having a substantially annular shape in the cross section.","Since the interface between the inner cladding and the outer core may be an interface where at least on one side there is a microstructure (the inner cladding region), the interface may not necessarily be seen as a smooth interface (e.g.","in the case of the two regions having identical background refractive indices).","Therefore, the shape of the outer core region may not necessarily be viewed as a smooth ring-like shape—hence the wording “substantially annular shape”; 4) a second cladding region surrounding the second core region, the second cladding region comprising a second cladding material of refractive index, N cl,2 , and the second cladding region having a substantially annular shape in the cross section.","In the present context, the wording “substantially annular shape” is used for the same reasons as discussed above.","In a preferred embodiment, the predetermined wavelength is in a range from 1.3 μm to 1.7 μm, such as from around 1.5 μm to 1.62 μm.","In another preferred embodiment, the optical fiber guides light at the predetermined wavelength in a higher-order mode.","This is preferred as a higher order mode in a fiber according to the present invention may exhibit a very high positive dispersion or a very high negative dispersion.","In a preferred embodiment, the present invention covers an article (this being a fiber optical communication system or parts thereof) that includes a module being employed for dispersion compensation in an optical communication system.","The module comprises an optical fiber according to the present invention.","In another preferred embodiment, the present invention covers an article that comprises a mode-converter that enables coupling from a conventional single mode fiber to an optical fiber according to the present invention.","Finally, it should be mentioned that the present invention covers both fibers that guide light by a modified version of total internal reflection as well as fibers that guide light by photonic band gap effects.","Glossary and Definitions For microstructures, a directly measurable quantity is the so-called “filling fraction” that is the volume of disposed features in a microstructure relative to the total volume of a microstructure.","For fibers that are invariant in the axial fiber direction, the filling fraction may be determined from direct inspection of the fiber cross-section, e.g.","using inspection methods known in the art.","In this application we distinguish between “refractive index”, “geometrical index” and “effective index”.","The refractive index is the conventional refractive index of a homogeneous material—naturally, this is also used to describe the refractive indices of the various materials themselves in a microstructured medium.","The geometrical index of a structure is the geometrically weighted refractive index of the structure.","As an example, a micro-structure consisting of air features that occupy 40% of the structure (the air features themselves having a refractive index=1.0) and a background material of silica (the silica background material having a refractive index ≈1.45) has a geometrical index of 0.4×1.0+0.6×1.45=1.27.The procedure of determining the effective refractive index, which for short is referred to as the effective index, of a given micro-structure at a given wavelength is well-known to those skilled in the art (see e.g., Joannopoulos et al., “Photonic Crystals”, Princeton University Press, 1995, or Broeng et al., Optical Fiber Technology, Vol.","5, pp.","305-330, 1999).","Usually, a numerical method capable of solving Maxwell's equation on full vectorial form is required for accurate determination of the effective indices of microstructures.","The present invention makes use of employing such a method that has been well documented in the literature (see previous Joannopoulos-reference).","In the long-wavelength regime, the effective index is roughly identical to the weighted average of the refractive indices of the constituents of the material, that is, the effective index is close to the geometrical index in this wavelength regime."],["1.BACKGROUND OF THE INVENTION The present invention relates to new designs of microstructured fibers providing improvements with respect to dispersion compensation (including dispersion slope compensation) for fiber optical communication systems and with respect to non-linear optical fibers for applications at near-infrared wavelengths.","THE TECHNICAL FIELD The development of optical amplifiers operating in the 1550 nm wavelength band of optical communication has during the past ten years formed the basis for a tremendous development of wavelength division multiplexed optical communication systems typically operating in the wavelength interval from 1530 nm to 1610 nm—and rapidly expanding.","These amplifiers have to a large degree removed the loss limitations of the optical communication links, and have paved the way for much longer transmission spans operating at significantly higher transmission bit rates (having a much higher transmission capacity).","At the same time the development of optical fiber amplifiers at first had the result that systems originally developed to operate in the 1310 nm wavelength band could be upgraded to the 1550 nm band with significant advantages —provided that the dispersion limitations of these systems could be compensated in an efficient manner.","To provide a picture of the possibilities and needs for dispersion compensation, it may be noted that when the bit rate is increased to 10 Gbit/s, the dispersion limits the transmission to around 50 km, which obviously raises the need for some sort of dispersion compensation.","As described by Grüner-Nielsen et al., ECOC'2000, pp.91-94, non-zero dispersion fibers (NZDF) seems to be the choice for future communication systems due to their low-dispersion and low-non-linear penalties.","For bit-rates of 10 Gbit/s NZDFs will not need dispersion compensation before some hundred kilometres of transmission.","However, in the future, when the bit rate increases to 40 Gbit/s, dispersion compensation will be needed already after approximately 5 kilometres for non-shifted fibers and after around 30 kilometres when NZDFs are used.","It is noteworthy that today several different types of optical transmission fibre are installed with various dispersion properties.","However, still a significant part of the installed optical fiber cables make use of non-shifted single-mode fibers, i.e.","optical fibres with zero dispersion wavelength at 1310 nm and a dispersion of about 17 ps/km/nm at the wavelength of 1550 nm.","The most established dispersion-compensation method today is the use of dispersion compensating fibers (DCFs) operating in the single-mode regime.","The use of dispersion compensating fibers was proposed by Lin, Kogelnik, and Cohen, Optics Letters, Vol.5, pp.476-478, 1980.The first DCFs were step index fibers, where the zero dispersion wavelength was moved to wavelengths above 1550 nm by increasing the core index and narrowing the core diameter (see Onishi et al., Electronics Letters, Vol.","30, pp.","161-163, 1994, and Bjarklev et al., Optics Letters, Vol.19, pp.","62-64, 1994).","These relatively simple fiber designs allow for control of the dispersion value at a given wavelength, but they do generally not provide the possibility of simultaneous control of the dispersion slope value.","It could be argued that one of the ultimate limits for the dispersion compensating properties of this simple step index fiber design may be obtained for a very thin rod of a high index material placed in vacuum (the low-index cladding).","At this point it is relevant to refer to the recent development within the area of fiber optics, where a large interest has been pointed towards so-called microstructured fibers, also known as photonic crystal fibers, photonic band gap fibers, hole-assisted fibres and holey fibers.","These are fibers providing a number of new properties that are of interest to a wide range of areas such as optical communications, sensor technology, spectroscopy, and medicine (see e.g.","Broeng et al., Optical Fiber Technology, Vol.","5, pp.","305-330, 1999; Broeng et al., Optics Communications, Vol.","156, pp.","240-244, 1998; Broeng et al, Optics Letters, Vol.","25, pp.","96-98, 2000; WO 99/64903; WO 99/64904; WO 00/60390; Birks et al., Electronics Letters, Vol.31, pp.1941-1943, October 1995; Knight et al., Journal of the Optical Society of America, A., Vol.15, pp.748-752, March 1998; Knight et al., Optical Materials Vol.","11, pp.","143-151, 1999; U.S. Pat.","No.","5,802,236; Monro et al., Journal of Lightwave Technology, Vol.","17, pp.","1093-1102, 1999; Ferrando et al., Optics Letters, Vol.","24, pp.","276-278, 1999; WO 00/06506).","The fibres are characterized by having a core surrounded by thin, parallel, voids/holes in a background material.","The background material is most often a single material such as e.g.","silica glass, and the voids/holes commonly contain air or vacuum, but they may also be filled with other glass materials, polymers, liquids, or gasses.","Depending on the application, the voids/holes may be periodically or randomly distributed, or they may be distributed in specially designed arrangements incorporating both periodic and non-periodic regions (see e.g.","WO 99/64903; WO 99/64904; WO 00/60390; U.S. Pat.","No.","5,802,236, Monro et al., Optics Letters, Vol.","25, pp.","206-208, 2000).","Now returning to the picture of a single rod of high-index material placed in vacuum, it should be noted that Birks et al.","(in IEEE Photonics Technology Letters, Vol.11, pp.","674-676, 1999) argued that properties of photonic crystal fibers (PCFs) could be modelled by a silica rod in air.","Such approximate calculations indicated that the dispersion of PCFs could exceed −2000 ps/km/nm—or they could compensate (to within + or −0.2%) of the dispersion of 35 times their length of standard fiber over a 100 nm wavelength range.","As already mentioned, it is a disadvantage of the step-index fibers that they cannot perform simultaneous compensation of dispersion and dispersion slope, and it is a further disadvantage that very high index contrasts are needed (either obtained through high co-dopant levels or by placing single rods in air).","The high-index contrast step-index fibers also have to be designed with very narrow core dimensions in order for the fibers to be single-moded.","Regarding non-linearity, this is a further disadvantage especially for systems operating at 40 Gb/s or higher.","After now having discussed the simplest approach of obtaining dispersion compensation in standard optical fibers and in photonic crystal fibers, it is useful to look at the further development of standard technology DCFs.","As we already have mentioned, one of the key limitations to the simple step-index design was the lack of simultaneous control of dispersion and dispersion slope.","One may argue that this limitation simply is due to a quite limited number of design parameters (for a step-index fiber only the core-cladding refractive index difference and the core radius may be selected).","The approach in obtaining more design flexibility dates back to the early days of standard fiber technology, and dispersion modification techniques were already studied in the early 1980'ies (see e.g., Monerie, IEEE Journal of Quantum Electronics, Vol.18, pp.535-542, 1982), where double-clad designs (also known as W-fibers) were evaluated.","Fibers of this double-clad type also have equivalents realised by photonic crystal fiber technology, and these microstructured fibers have also been explored for dispersion compensating applications.","The first microstructured fibers for such applications were disclosed by DiGiovanni et al.","(see EP 0 810 453 and U.S. Pat.","No.","5,802,236).","DiGiovanni et al.","discussed how microstructured fibers with a cladding divided into two regions could provide improved dispersion characteristics compared to conventional fibers.","DiGiovanni et al.","used microstructured fibers with an inner, microstructured cladding region having a lower effective refractive index compared to an outer, microstructured cladding region to obtain a dispersion of more than −1500 ps/nm/km.","The fibers disclosed by DiGiovanni et al.","comprise a single core region (preferably in the centre of the fiber), and it is stated in U.S. Pat.","No.","5,802,236 that “In general, the microstructure cladding features should be disposed such that the cladding region does not contain any matrix regions of extend in the x-y plane sufficient to act as a secondary core, i.e.","to support a propagating radiation mode”.","The present inventors have, however, realised that it is advantageous to design microstructured fibres that have at least one region that can support one or more propagating modes other than the mode(s) supported in the central core region.","Hence, such a region (that has larger dimensions than the central core and an effective refractive index higher than (No+Nc)/2, where No is the core refractive index and Nc is the effective refractive index of the cladding) that may act as a second core is preferred.","In fact, the present inventors have realized that coupling between a mode in the central core region and a mode in the second core is highly advantageous for microstructured optical fibers with special dispersion properties.","It is further important to notice that the fiber may be operated to guide light only in a single of the cores—preferably the central core.","This desired mechanism may also be described as avoided “crossing” of the core modes, as shall be discussed at a later stage in the detailed description of the invention.","It should at this point be noted that strong dispersion compensation generally is obtained by spectrally shifting the effective refractive index of the guided mode(s).","This requires refractive index profile control on a sub-wavelength scale, and the effect is closely related to the spatial redistribution of the optical mode as a function of wavelength.","When this spatial redistribution of the guided mode takes place in a W-fiber design, the guided mode shifts (for increasing wavelengths) from the high index core to the cladding.","This is an effect that generally ties the dispersion compensating effect strongly together with a strong leakage loss and macro bending loss sensitivity very near the desired wavelength for dispersion compensation.","Consequently, W-designs have to be made for an extremely narrow selection of production parameters, and this fiber design, therefore, becomes less attractive than designs, where the spatial distribution of optical power is shifted within guided regions of the fiber core.","This is also the reason why standard fiber technology generally utilises triple-cladding designs for dispersion compensation (see e.g., Vengsarkar et al., OFC'93, San Jose, USA, Feb. 21-26, 1993, Post deadline paper PD13; Antos, and Smith, IEEE Journal of Lightwave Technology; vol.12, pp.","1739-1745, 1994; Vengsarkar et al., OFC'94, paper ThK2, pp.","222-227, 1994; Onishi et al., ECOC'94, pp.681-684,1994; Akasaka et al., OFC'96, paper ThA3, pp.","201-202,1996).","These designs have several advantages over the step index design—among others is negative dispersion slope to provide partly or full dispersion slope compensation.","Using standard fiber technology, triple-cladding designs have a narrow high-delta core (i.e.","highly doped to obtain a large refractive index difference compared to pure silica) surrounded by a deeply depressed cladding followed by a raised cladding (we here mention the different elements in an order starting at the centre of the fiber cross-section and stepping outwards in radial direction).","The third cladding layer is generally made of pure silica.","It should also be noted that in order to suppress higher-order mode guidance (lowering the cut-off wavelength of the DCF) an additional depressed cladding layer is often introduced between the raised cladding layer and the outermost cladding.","However, for simplicity (and because its influence on the fundamental mode properties often are limited) this fourth cladding layer is often not discussed in large detail.","As we have discussed with the more simple standard fiber designs, also triple-cladding design equivalents using microstructured—or PCF—technology have been considered by Fajardo et al.","WO 00/16141.Fajardo et al.","describe an optical waveguide in which the density and thus the effective refractive indices of the cladding layer is caused to change in a pre-selected way axially along the fiber.","The axial change in density of the cladding layer is due to the fraction of the cladding volume that is air or a glass composition different from that of the base cladding glass.","Fajardo et al.","argue that the axial variation in clad indices changes the signal mode power distribution, thereby changing key waveguide fiber parameters such as magnitude and sign of dispersion, cut-off wavelength and zero dispersion wavelength along the fiber length.","It is a disadvantage of the dispersion compensating fibers using standard fiber fabrication techniques (i.e., fibers fabricated from doped silica) that limited dopant levels (given by internal stress, material deposition efficiencies etc.)","does not provide as large (negative) dispersion values as other competing techniques.","It is a further disadvantage that standard single-mode DCFs typically have significantly smaller spotsize values than the transmission fibers, whose dispersion they are to compensate.","The spotsize mismatch generally leads to significant coupling losses, and/or complicated splicing techniques including intermediate fibers, special techniques of material diffusion control etc.","When a DCF is used for discrete compensation at the terminal sites, one of the drawbacks of DCFs is that the total link attenuation is increased.","The added attenuation must be compensated with additional gain in amplifiers, which degrades the signal-to-noise ratio and increases the cost of the system.","To minimise this problem, the DCF should have as high a negative dispersion, DDCF, as possible and as low attenuation, αDCF, as possible.","Therefore, a figure of merit (FOM) for dispersion compensating fibers may be defined as the numerical value of DDCF/αDCF.","Typical FOM values of presently employed DCFs is around 200 ps/nm/dB, where the attenuation is around 0.5 dB/km and the dispersion is around −100 ps/nm/km.","Hence, for a fiber according to the present invention providing a dispersion of −400 ps/nm/km or more negative, loss issues are less restricting than for conventional DCFs—or DCFs according to the present invention may provide higher FOM for similar loss levels.","For a dispersion compensating fiber according to the present invention that provides for example a dispersion of −1000 ps/nm/km or more negative, an improved FOM may be achieved for fiber attenuation of up to 5 dB/km compared to conventional DCFs.","It should be noted that dispersion compensating fibers fabricated in large-scale production using standard fiber technology normally are designed to have a dispersion of about −100 ps/km/nm at the wavelength of 1550 nm—and having FOM in the order of 200 ps/nm/dB (see e.g., Grüner-Nielsen et al.","ECOC'2000, Munich, pp.","91-94).","However, more negative dispersion values may be obtained—still having good FOM values, and as an example the work of Knudsen et al., LEOS'2000 annual meeting, Paper TuZ2, pp.338-339 may be mentioned.","This works demonstrated a dispersion of −190 ps/km/nm for a FOM of 307 ps/nm/dB at 1550 nm.","Considering the background of dispersion compensation through the use of specially designed fibers it also becomes relevant to examine the dispersion compensating fibers described by Thyagarajan et al., IEEE Photonics Technology Letters, Vol.8, pp.","1510-1512, 1996.In this letter, Thyagarajan et al.","describe how very large negative dispersion values (as low as −5100 ps/km/nm) are predicted for fibers consisting of two concentric cores.","The desired dispersion compensation is obtained using a centrally placed high-index core (from 2-2.5% relative refractive index difference) and a ring shaped core, chosen independently so that each of the cores supports a single azimuthally symmetric propagating mode at a wavelength of 1550 nm.","The idea described by Thyagarajan et al.","is that the two-core fiber design then will support two azimuthally symmetric modes (similar to two so-called super modes of a directional coupler), and that these modes are nearly phase matched at 1550 nm.","Close to this phase matching wavelength, the mode index of the composite core fiber changes rapidly due to a strong coupling between the two individual modes of the inner core and outer core.","Due to a strong refractive index asymmetry between the two cores, there is a rapid change in the slope of the wavelength variation of the fundamental mode index (of the composite structure) leading to a large dispersion around 1550 nm.","A fiber of this dual concentric core type has been fabricated, and dispersion results as low as −1800 ps/km/nm at the wavelength of 1550 nm has recently been presented by Auguste et al., IEE Electronics Letters, Vol.","36, pp.","1689-1691, 2000.Also in this work the functionality of the fiber is described through the coupling between two super modes—one of the super modes having a large negative dispersion and the other having a large positive dispersion—and light being guided through the fiber only in the super mode having negative dispersion.","Note also that fibers of this type has been further described by Auguste et al., Optics Communications, Vol.","178, pp.","71-77, 2000, where the influence of a central dip—generally present in fibers fabricated by the modified chemical vapour deposition (MCVD) method—is studied and demonstrated to be of importance.","Considering these dual concentric core fiber designs, it is important to note that the fiber propagation properties also may be interpreted by a more ordinary mode description using linearly polarised (LP) modes in optical fibers.","If we look at the dual concentric core designs from this point of view, we find that the LP01 mode (representing one of the previously mentioned super modes) shows a strong negative dispersion.","At the same time, the LP02 mode (representing the other of the previously mentioned super modes) shows a strong positive dispersion, and more important—the fiber supports guidance of the LP02-mode.","Looking closer at the fiber designs presented by Thyagarajan et al., one also finds that the LP11 mode is strongly guided by the fiber structure.","It may be a disadvantage of the dual concentric core designs fabricated by standard fiber technology that the fibers strongly guide several modes.","It is a further disadvantage of the dual concentric core designs fabricated by standard fiber technology that very high co-dopant concentrations generally are needed, since it may result in highly increased fiber attenuation, and moreover, it is difficult to maintain a homogeneous, and reproducible dopant level.","The latter issue is indicated by Auguste et al.","(Optics Communications, Vol.","178, pp.","71-77, 2000), where it is described how the well known central index dip in MCVD fabricated fibers may introduce strong variations in the spectral location of the dispersion peak (more than 100 nm shifting was reported).","It is yet a further disadvantage of the standard technology dual concentric core fibers that the limited index contrast in doped glass technology makes it necessary to operate with ring shaped cores of rather wide outer radii (fibers presented by Thyagarajan et al.","and Auguste et al.","have radii of the outermost core near 20 microns).","This leads to a strongly spatially extended fundamental mode field for wavelengths above 1550 nm, which eventually may result in significant macro-bending sensitivity.","High dopant levels are not necessary in PCFs, because index contrast may be obtained through microstructuring.","Therefore, the parameter sensitivity may be reduced, and fibers with different guiding properties between the guided and higher order modes may be obtained (mode discrimination is possible).","Through a wider range of design possibilities using PCF technology, we may solve the bending loss problem, and at the same time obtain a larger negative dispersion.","It is an object of the present invention to provide improved designs and design parameters for microstructured fibers employed for dispersion compensating applications (including dispersion slope compensation).","The improvements relates to strong dispersion, control of dispersion slope, low bending losses, large mode-field diameters, lower sensitivity on parameter fluctuations—allowing better reproducibility compared to prior art fibers, and lower requirements on doping levels of silica.","Photonic crystal fibres (PCFs) having multiple cladding layers have been described by Hasegawa et al.","in EP 1 118 887 A2.Hasegawa et al.","claim an optical fiber including a core region and cladding regions of not less than three layers, which surround the core region in order, wherein at least one of the cladding regions has lower mean refractive index than both adjacent cladding regions, and at least one cladding region is provided with a plurality of sub medium regions each having a refractive index lower than a main medium constituting this cladding region.","The present inventors have, however, realised that it is advantageous to design microstructured fibers that have at least one region that can support one or more propagating modes other than the mode(s) supported in the central core region.","It is a disadvantage of the fibers described by Hasegawa et al.","that they do not include such a region that may act as a core in its own right supporting a propagating mode, since it may limit the possible magnitude of the obtainable dispersion.","Furthermore, the present inventors have realised that it may be advantageous not to have a cladding region with a lower effective refractive index than two adjacent cladding regions.","In fact the present inventors have realised that it is an advantage to have a region with a higher effective refractive index than two adjacent cladding—such a region is here termed “second core”.","The fibres disclosed by Hasegawa et al.","exhibit a negative dispersion of around −200 ps/km/nm, whereas the micro-structured fibers disclosed by the present inventors provides negative dispersion in excess of −10000 ps/nm/km.","In the patent application by Hasegawa et al., it is given that the mean refractive index n3 of the outer cladding region obeys the relation n2>n3, where n2 is the refractive index of the second inner cladding.","Hasegawa et al.","obtains this relation by incorporating low-index features in cladding region 3.This is in contrast to a number of preferred embodiments of the present invention, where the outer cladding consists of a homogeneous material.","SUMMARY OF THE INVENTION In a first aspect, the present invention relates to an optical fiber having an axial direction and a cross section perpendicular to said axial direction.","The optical fiber has negative dispersion in the vicinity of a predetermined wavelength.","The optical fiber is characterised by a number of regions: 1) a first core region being positioned substantially in a center of the optical fiber, the first core region comprising a first core material of refractive index, Nco,1.In the present context, “positioned substantially” means that it is desired to have the first core placed in the center, but manners of production may introduce minor structural deviations such that a center of the first core region and a center of the whole optical fiber may not coincide.","Also, the optical fiber may have a non-circular outer shape in the cross section, making the definition of a center of the optical fiber less stringent.","In the case of a non-circular outer shape, the center is determined as the center-of-gravity of a two-dimensional element with the same outer shape as the optical fiber.","2) a first cladding region surrounding the first core region, the first cladding region comprises a multiplicity of spaced apart first cladding features that are elongated in the axial direction and disposed in the first cladding material, said first cladding material having a refractive index Ncl,1; 3) a second core region surrounding the first cladding region, the second core region comprising a second core material of refractive index, Nco,2, and the second core region having a substantially annular shape in the cross section.","Since the interface between the inner cladding and the outer core may be an interface where at least on one side there is a microstructure (the inner cladding region), the interface may not necessarily be seen as a smooth interface (e.g.","in the case of the two regions having identical background refractive indices).","Therefore, the shape of the outer core region may not necessarily be a viewed as a smooth ring-like shape—hence the wording “substantially annular shape”; 4) a second cladding region surrounding the second core region, the second cladding region comprising a second cladding material of refractive index, Ncl,2, and the second cladding region having a substantially annular shape in the cross section.","In the present context, the wording “substantially annular shape” is used for the same reasons as discussed above.","Typically, the predetermined wavelength is in a range from 1.3 μm to 1.7 μm, such as from around 1.5 μm to 1.62 μm.","In a preferred embodiment, the optical fiber has a higher refractive index in the first core region than in the second core region.","In another preferred embodiment, the optical fiber has identical refractive index in the first core region and in the second core region.","This allows for example to manufacture the fiber in a single material, such as e.g.","pure silica.","In a preferred embodiment, the optical fiber has a lower refractive index in the first core region than in the second core region.","This allows for example to have a very large mode field diameter.","In another preferred embodiment, the first cladding region has a substantially annular shape in the cross section.","The wording “substantially” being used for the same reasons as previously discussed.","In another preferred embodiment, the optical fiber has a first inscribed core diameter, Dco,1, being larger than 3 times said predetermined wavelength, such as larger than 5 times said predetermined wavelength, such as larger than 7 times said predetermined wavelength, such as larger than 10 times said predetermined wavelength.","In another preferred embodiment, Dco,1 is in the range from 4 μm to 25 μm, such as in the range from 4.0 μm to 5.0 μm, such as in the range from 5.0 μm to 6.5 μm, such as in the range from 6.5 μm to 10.0 μm, such as in the range from 10.0 μm to 25.0 μm.","In another preferred embodiment, the first core region has a varying refractive index profile, said varying refractive index profile having a highest refractive index equal to Nco,1, and said varying index profile being characterised by an □-profile, where □ is in the range from 0 to 100, such as □ equal to 2, 3 or higher.","In another preferred embodiment, the second core region is a homogeneous region.","In another preferred embodiment, the second core region is microstructured.","This increases the effective refractive index ranges that are available for the second core region without the use of doping.","In another preferred embodiment, the microstructured features in the second core are positioned substantially circularly symmetric with respect to a center of said first core region.","This is preferred in order to lower polarisation mode dispersion (PMD) in the fiber.","An optical fiber according to any one of the preceding claims, wherein said second core features are arranged in a single layer surrounding said first core region, such that a distance from a second core feature to a center of said first core region is substantially identical for all second core features.","This is preferred in order to lower PMD, and further preferred for ease of fabrication.","In another preferred embodiment, the second core features are arranged in two or more layers surrounding said first core region.","This is preferred to reduce bending losses.","In another preferred embodiment, the number of second core features is equal to or larger than 6, such as equal to or larger than 18.A high number is preferred to modify the effective refractive index of the second core region.","In another preferred embodiment, the second core features have a diameter Dco,2 and a center-to-center spacing between nearest second core features of Λco,2, and Dco,2/Λco,2 is in the range from 0.01 to 0.5, such as from 0.1 to 0.2.Generally, the microstructured features in the second core should be relatively small in order to ensure the possibility of the second core to support a limited number of modes.","In another preferred embodiment, Dco,2 is in the range from 0.1 μm to 5 μm.","Typically, this is the design parameter range that provides strongest dispersion.","In another preferred embodiment, λ/Λco,2 is in the range from 0.2 μm to 20 μm.","Typically, this is the design parameter range that provides strongest dispersion.","In another preferred embodiment, the first cladding features are positioned substantially circularly symmetric with respect to a center of said first core region.","This is preferred to lower PMD in the fiber.","In another preferred embodiment, the first cladding features are arranged in a single layer surrounding said first core region, such that a distance from a first cladding feature to a center of said first core region is substantially identical for all first cladding features.","This is preferred to lower PMD in the fiber.","In another preferred embodiment, the said first cladding features are arranged in two or more layers surrounding said first core region.","This is preferred to lower bending losses in the fibers and/or to provide stronger dispersion.","In another preferred embodiment, the number of said first cladding features is equal to or larger than 3, such as equal to or larger than 6, such as equal to or larger than 8, such as equal to or larger than 18.This range of numbers of features allows for tailoring of the birefringence of the fiber to a large degree.","Higher numbers of features may be desired to provide low PMD for example through more circular symmetric feature arrangements.","In another preferred embodiment, the first cladding features have a diameter Dcl,1 and a center-to-center spacing between nearest first cladding features of Λcl,1, and Dcl,1/Λcl,1 is in the range from 0.2 to 0.8, such as from 0.4 to 0.6.To provide strong dispersion, relatively large features are required.","Hence, the features should be large enough to provide a buffer region between the two cores.","In another preferred embodiment, Dcl,1 is in the range from 0.1 μm to 10 μm.","Typically, this range provides the most attractive design parameter range in order to obtain strong dispersion for wavelengths around 1.55 μm.","In another preferred embodiment, Λcl,1 is in the range from 0.2 μm to 20 μm.","Typically, this range provides the most attractive design parameter range in order to obtain strong dispersion for wavelengths around 1.55 μm.","In another preferred embodiment, Dcl,1/Λcl,1 is larger than Dco,2/Λco,2.This is preferred to ensure that the first cladding region acts as a buffer between the two cores.","In another preferred embodiment, the second cladding region comprises a multiplicity of spaced apart second cladding features that are elongated in the axial direction and disposed in said second cladding material.","This provides further means for tailoring the dispersion properties of the fiber.","In another preferred embodiment, second cladding features have a diameter Dcl,2 and a center-to-center spacing between nearest second cladding features of □cl,2, where Dcl,2/Λcl,2 is in the range from 0.01 to 0.5, such as from 0.1 to 0.2.It is often preferred that these features are comparable to the features of the second core.","It may be preferred that the second core is only weakly guiding or has a cut-off close to or around the operational wavelength.","Hence the second cladding features may be preferred to have a limited size in order to not provide strong confinement for the second core—or alternatively not to provide confinement of a high number of modes in the second core.","In another preferred embodiment, Dcl,2 is in the range from 0.1 μm to 5 μm.","In accordance with the above-stated preferred embodiment, this is preferred to have the second cladding features being comparable in size to the second core features.","Similarly, it is further preferred that Λcl,2 is in the range from 0.2 μm to 20 μm.","In another preferred embodiment, Λcl,1 is larger than Λcl,2.This is preferred in order to allow the first and second cladding regions to have the same background material, but the effective refractive index of the two regions to be different at a given wavelength, such as the predetermined wavelength.","In particular, it is preferred to have Λcl,1 larger or equal to 3Λcl,2 as such a ratio may easily be realised using well known methods for fabricating microstructured fibers, such as methods employing stacking of capillary tubes.","In another preferred embodiment, Λcl,1 is substantially identical to Λcl,2.This may be preferred as similar pitches (i.e.","center-to-center features spacing or lattice period in the case of cladding comprising periodic feature arrangements) often allow simple fabrication, thereby improving reproducibility of the fiber.","In another preferred embodiment, Dcl,1 is larger than Dcl,2.This is preferred in order for the first cladding region to have a lower effective, refractive index than the second cladding.","In another preferred embodiment, Dcl,1/Λcl,1 and Dcl,2/Λcl,2 are substantially identical.","This may be preferred as similar hole and pitch sizes provide the simplest fabrication, thereby improving reproducibility of the fiber.","In another preferred embodiment, the optical fiber comprises one or more glass materials.","In another preferred embodiment, the optical fiber comprises silica.","In another preferred embodiment, the optical fiber comprises polymer.","In another preferred embodiment, the first cladding features, second core features, and/or second cladding features are voids comprising vacuum, air, or another gas.","In another preferred embodiment, the optical fiber has a non-circular shape of the outer cladding, such as a non-polygonal shape, such as an elliptical shape.","This may be advantageous for a range of reasons—for example for fibers with high birefringence (for example for handling splicing issues), or to ensure bending in certain preferred directions.","In another preferred embodiment, the outer shape has a predetermined orientation in the cross section, the predetermined orientation being determined from the position of said first cladding features.","This may be preferred to further ensure bending in certain preferred directions with respect to the actual directions in the microstructure in and/or around the core region.","Also for polarisation maintaining, dispersion compensating fibers this is preferred.","In another preferred embodiment, the first core region has a non-circular shape in the cross section, such as an elliptical shape.","This is preferred to provide a high birefringence in the optical fiber.","In another preferred embodiment, the first cladding region has a non-circular, inner shape in the cross section, such as an elliptical, inner shape, providing a high birefringence in the optical fiber.","This is further preferred for polarisation maintaining, dispersion compensating fiber applications.","In another preferred embodiment, the optical fiber guides light at predetermined wavelength in a single mode.","In another preferred embodiment, the optical fiber guides light at predetermined wavelength in a higher order mode.","This is preferred as higher order modes may provide even stronger dispersion than the fundamental mode including the possibility of strong positive dispersion.","In another preferred embodiment, the invention covers an article comprising a module, where the module is employed for dispersion compensation in an optical communication system, and the module comprising an optical fiber according to the invention.","In a second aspect, the present invention relates to an optical fiber having an axial direction and a cross section perpendicular to said axial direction.","The optical fiber has positive dispersion in the vicinity of a predetermined wavelength.","The optical fiber is characterised by a number of regions: 1) a first core region being positioned substantially in a center of the optical fiber, the first core region comprising a first core material of refractive index, Nco,1.In the present context, “positioned substantially” means that it is desired to have the first core placed in the center, but manners of production may introduce minor structural deviations such that a center of the first core region and a center of the whole optical fiber may not coincide.","Also, the optical fiber may have a non-circular outer shape in the cross section, making the definition of a center of the optical fiber less stringent.","In the case of a non-circular outer shape, the center is determined as the center-of-gravity of a two-dimensional element with the same outer shape as the optical fiber.","2) a first cladding region surrounding the first core region, the first cladding region comprises a multiplicity of spaced apart first cladding features that are elongated in the axial direction and disposed in the first cladding material, said first cladding material having a refractive index Ncl,1; 3) a second core region surrounding the first cladding region, the second core region comprising a second core material of refractive index, Nco,2, and the second core region having a substantially annular shape in the cross section.","Since the interface between the inner cladding and the outer core may be an interface where at least on one side there is a microstructure (the inner cladding region), the interface may not necessarily be seen as a smooth interface (e.g.","in the case of the two regions having identical background refractive indices).","Therefore, the shape of the outer core region may not necessarily be viewed as a smooth ring-like shape—hence the wording “substantially annular shape”; 4) a second cladding region surrounding the second core region, the second cladding region comprising a second cladding material of refractive index, Ncl,2, and the second cladding region having a substantially annular shape in the cross section.","In the present context, the wording “substantially annular shape” is used for the same reasons as discussed above.","In a preferred embodiment, the predetermined wavelength is in a range from 1.3 μm to 1.7 μm, such as from around 1.5 μm to 1.62 μm.","In another preferred embodiment, the optical fiber guides light at the predetermined wavelength in a higher-order mode.","This is preferred as a higher order mode in a fiber according to the present invention may exhibit a very high positive dispersion or a very high negative dispersion.","In a preferred embodiment, the present invention covers an article (this being a fiber optical communication system or parts thereof) that includes a module being employed for dispersion compensation in an optical communication system.","The module comprises an optical fiber according to the present invention.","In another preferred embodiment, the present invention covers an article that comprises a mode-converter that enables coupling from a conventional single mode fiber to an optical fiber according to the present invention.","Finally, it should be mentioned that the present invention covers both fibers that guide light by a modified version of total internal reflection as well as fibers that guide light by photonic band gap effects.","Glossary and Definitions For microstructures, a directly measurable quantity is the so-called “filling fraction” that is the volume of disposed features in a microstructure relative to the total volume of a microstructure.","For fibers that are invariant in the axial fiber direction, the filling fraction may be determined from direct inspection of the fiber cross-section, e.g.","using inspection methods known in the art.","In this application we distinguish between “refractive index”, “geometrical index” and “effective index”.","The refractive index is the conventional refractive index of a homogeneous material—naturally, this is also used to describe the refractive indices of the various materials themselves in a microstructured medium.","The geometrical index of a structure is the geometrically weighted refractive index of the structure.","As an example, a micro-structure consisting of air features that occupy 40% of the structure (the air features themselves having a refractive index=1.0) and a background material of silica (the silica background material having a refractive index ≈1.45) has a geometrical index of 0.4×1.0+0.6×1.45=1.27.The procedure of determining the effective refractive index, which for short is referred to as the effective index, of a given micro-structure at a given wavelength is well-known to those skilled in the art (see e.g., Joannopoulos et al., “Photonic Crystals”, Princeton University Press, 1995, or Broeng et al., Optical Fiber Technology, Vol.","5, pp.","305-330, 1999).","Usually, a numerical method capable of solving Maxwell's equation on full vectorial form is required for accurate determination of the effective indices of microstructures.","The present invention makes use of employing such a method that has been well documented in the literature (see previous Joannopoulos-reference).","In the long-wavelength regime, the effective index is roughly identical to the weighted average of the refractive indices of the constituents of the material, that is, the effective index is close to the geometrical index in this wavelength regime.","3.BRIEF DESCRIPTION OF THE DRAWINGS In the following, by way of examples only, the invention is further disclosed with detailed description of preferred embodiments.","Reference is made to the drawings in which: FIG.","1a (prior art) shows schematically the refractive index profile of a conventional step-index fiber.","FIG.","1b (prior art) shows schematically the cross-section of an equivalent index-guiding photonic crystal fiber.","FIG.","2a (prior art) shows schematically the refractive index profile of a conventional W-type fiber generally suggested for dispersion management.","FIG.","2b (prior art) shows schematically the cross-section of a microstructured fiber with two different air-void dimensions used to obtain an effective index profile of the W-type.","FIG.","3a (prior art) shows schematically the refractive index profile of a conventional triple-clad fiber of a type generally used for standard dispersion compensating fibers.","FIG.","3b (prior art) shows schematically the cross-section of a microstructured fiber with three different air-void dimensions used to obtain an effective index profile of the triple-clad type.","FIG.","4 (prior art) shows the effective index of micro-structures comprising air holes as a function of normalised wavelength for different air hole dimensions for.","FIG.","5 (prior art) shows schematically the cross-section of a microstructured fiber for dispersion compensation, the fiber having a doped central core surrounded by an annular inner cladding section containing larger air holes than those used to form the microstructured outer cladding; The fiber comprises further a high-index features in the center of the core.","FIG.","6 (prior art) shows the effective mode indices of periodic outer cladding regions and of a hypothetical honeycomb structures shown in FIG.","5 (representing the central fiber region) as a function of the normalised wavelength.","Curves are shown for honeycomb structures with different air-hole dimensions.","FIG.","7 shows the effective mode indices of periodic outer cladding regions and of a hypothetical honeycomb structures (representing the central fiber region) as a function of the normalised wavelength.","Curves are shown for different doping levels in the central core, and for different honeycomb air-hole dimensions.","FIG.","8 (embodiment) shows schematically the cross-section of microstructured fiber for dispersion compensation, the fiber having a doped central core surrounded by an annular inner cladding section containing larger air holes than those used to form the microstructured outer cladding.","Between the inner and outer cladding regions, a higher effective index is obtained in an annular segment, where air holes have been removed (compared to the outer cladding).","FIG.","9 shows a simulation of the mode indices for a fiber according to the present invention.","FIG.","10 shows a simulation of the dispersion of a dispersion-compensating fiber according to the present invention.","FIG.","11 shows the field distribution of the fundamental mode for a microstructured fiber according to the invention.","FIG.","12 shows schematically a fiber according to the present invention where the outer cladding is homogeneous and a) the outer core comprises high-index features or b) the outer core comprises an annular high-index region.","FIG.","13 shows qualitatively the operation of a dispersion compensation fiber according to the present invention that may have large negative dispersion and low bending losses.","FIG.","14 shows schematically a preferred positioning of low-index features in the inner cladding and of high-index features in the outer core with respect to each other.","FIG.","15 shows schematically a fiber according to the present invention where the outer cladding is homogeneous and comprises a material different from the background material of the inner cladding and of the outer core.","Typically, the outer cladding material has a lower refractive index than the background material of the inner cladding and of the outer core.","FIG.","16 shows a simulation of the mode indices for another fiber according to the present invention.","FIG.","17 shows a simulation of the dispersion of a range of fibers according to the present invention.","FIG.","18 shows a simulation of the wavelength of strongest negative dispersion and the corresponding hole spacing for a range of fibers according to the present invention designed for operation around 1550 nm.","FIG.","19 shows a simulation of the strongest negative dispersion value of a range of fibers according to the present invention.","FIG.","20 shows a simulation of the width of the dispersion “dip” of a range of fibers according to the present invention.","FIG.","21 shows a simulation of the dispersion and relative dispersion slope for a fiber according to the present invention.","FIG.","22 shows a simulation of the dispersion and relative dispersion slope for another fiber according to the present invention.","FIG.","23 shows a simulation of the dispersion and relative dispersion slope for yet another fiber according to the present invention.","FIG.","24 illustrates the operation of another fiber according to the present invention.","The figure illustrates the possibility of using higher order modes in the central core for dispersion compensating applications for fibers according to the present invention.","FIG.","25 shows a schematic example of another fiber according to the present invention.","FIG.","26 illustrates the operation of a fiber according to the present invention having an outer core with the same refractive index as the refractive index of the inner cladding region.","FIG.","27 illustrates the operation of a fiber according to the present invention being comparable to the fiber in the previous figure, but with a larger size of the outer core.","FIG.","28 shows a simulation of the dispersion and the effective area of the fiber of the previous figure.","FIG.","29 shows a simulation of the dispersion and the effective area of a fiber according to the present invention having a high-index core not being in contact with the inner cladding.","FIG.","30 shows a comparison of the effective area of the fibers in the two previous figures.","FIG.","31 shows a comparison of the dispersion of the fibers in FIGS.","28 and 29.FIG.","32 shows schematically the cross-section of a micro-structured fiber for dispersion compensation, the fiber has a core with a doped feature, the core being surrounded by an annular inner cladding section containing more closely placed air holes than those used to form the microstructured outer cladding.","Between the inner and outer cladding regions, a higher effective index is obtained in an annular segment, where air holes have been removed (compared to the outer cladding).","FIG.","33 shows schematically the cross-section of a micro-structured fiber for dispersion compensation, the fiber having a core with a doped portion and the core is surrounded by an annular inner cladding section containing more closely placed air holes than those used to form the microstructured outer cladding.","Compared to the previous figure, additional air holes have been removed in the outer core region.","FIG.","34 schematically shows the cross-section of a micro-structured fiber according to the invention, in which the outer core comprises high-index features.","FIG.","35 schematically shows the cross-section of a micro-structured fiber according to the invention, in which the outer core comprises a single annular feature with a higher refractive index than the outer cladding background material.","FIG.","36 schematically shows the cross-section of a micro-structured fiber according to the invention, in which the outer cladding is formed by air holes placed in concentric circles.","FIG.","37 schematically shows the cross-section of a micro-structured fiber according to the invention, in which the outer cladding is formed by air-holes placed in concentric circles, and where the inner cladding is formed by air holes of a non-circular cross section.","a) Four glass segments are used to keep the central part of the fiber in place.","b) Three glass segments are used to keep the central part of the fiber in place.","FIG.","38 shows schematically the effective index variation of microstructures that may be utilised for realising fibers according to the present invention with a core comprising material with a maximum refractive index being lower than a highest refractive index of any material in the remainder of the fiber.","FIG.","39 schematically shows the cross-section of a micro-structured photonic band gap fiber according to the invention.","A honeycomb-structured cladding has been used, and the core is formed by an additional hole.","Surrounding the central hole is a doped region.","FIG.","40 schematically shows the cross-section of a micro-structured photonic band gap fibre according to the invention.","The fiber contains an annular ring with a number of additional air holes placed to enhance the dispersion compensating properties of the PBG fiber, and the outer core comprises small low-index features.","FIG.","41 illustrates the effective index of a band gap guided mode in a fiber structure according to the invention.","FIG.","42 shows a simulation of the inverse group velocity dispersion that may be obtained for fibers according to the present invention.","FIG.","43 shows microscope pictures of a real fiber according to the present invention.","The figure further shows the dispersion characteristics of the fiber at wavelengths around 1550 nm.","FIGS.","44a-44c show microscope pictures of another real fiber according to the present invention.","The figure further shows the dispersion characteristics of the fiber at wavelengths around 1550 nm.","FIGS.","45a-45c show another example of a real fiber according to the present invention.","FIGS.","46a-46b provide a schematic example of a fiber according to the present invention.","FIGS.","47a-47b provide a similar schematic example.","4.DETAILED DESCRIPTION In FIG.","1a, the index profile of standard step-index fiber design according to prior-art is illustrated.","In this type of fiber design, the core region 101 is generally formed by a narrow circular section, which is strongly co-doped by germanium to provide the raised index in comparison with the surrounding cladding region 102.It should, however, be noted that the fiber also may be realised by an un-doped (pure silica) core surrounded by a doped cladding region (typically F-doped silica).","The design also includes a thin solid rod of glass placed in air (or vacuum) or even another lower index material compared to the core glass.","In FIG.","1b, the equivalent microstructured fiber is shown schematically.","In this realisation, an un-doped core 103 is surrounded by a cladding 104 that is formed by placing air holes 105- or voids—in a silica background material.","This realisation of the cladding provides an average refractive cladding index, which is lower than the refractive index of the core.","The inscribed, dashed circle 106 in FIG.","1b indicates one division between the core region and the cladding region—defining the core diameter, Dco, as the inscribed diameter.","In FIG.","2a, the refractive index profile of a prior art fiber of the so-called w-profile type is illustrated.","This type of refractive index is characterised by a core 201—typically formed by an index raised by germanium co-doping.","The core is surrounded by a depressed (typically through flour co-doping) cladding region 202, and this inner cladding region is surrounded by an outer cladding 203, which typically is realised in pure silica.","As described in the background description of the invention, these fibers operate by a spectrally dependent effective mode index, which is related to spectrally dependent power redistribution of the fundamental mode.","FIG.","2b (prior art) illustrates an example of a micro-structured fiber realised to form an equivalent to the W-profile fiber of standard technology.","Here, the core is formed by a solid core region 204 surrounded by an inner cladding region 205 in which air holes 206 is placed.","Since the air holes 206 of the inner cladding region have a larger cross section than the air holes 208 of the outer cladding 207, the effective index of the inner cladding becomes smaller than that of the outer cladding.","Generally, this type of W-profile fiber will show significant macro-bending losses near the wavelength where a significant negative dispersion is obtained.","In FIG.","3a (prior art), the index profile of a triple-cladding standard optical fiber for dispersion compensation (DCF) is illustrated.","A DCF of this type has a narrow highly-doped core 301 (typically germanium is used as index raising dopant) surrounded by a depressed cladding 302 (typically flour-doped to refractive index levels in the order of −0.5%) This germanium doping in the central part of the fiber may typically result in an increase of the refractive index of 2-3% using rather narrow core radii in the order of 1-2 μm.","The highly doped core is favourable as it enables a high negative dispersion, but the drawback is increased attenuation (as described by Grüner-Nielsen et al., ECOC'2000, pp.91-94).","The depressed cladding decreases the dispersion and is necessary to obtain negative dispersion slope around 1550 nm.","Another index ring 303 with raised index compared to the cladding is often introduced to improve bending properties, but the refractive index is normally not raised much above 0.5%, since a high index value will lead to multimode operation of the fiber.","Outermost the fiber has a pure silica cladding 304.FIG.","3b (prior art) illustrates the transversal cross section of a microstructured optical fiber designed to make use of similar (but enhanced) properties compared to standard fiber technology.","The illustration shows the core 305 formed by a solid material.","The core is surrounded by an inner cladding 306 in which air holes having relatively large cross sections 307 are placed to form an effective depressed cladding.","The inner cladding is, furthermore, surrounded by a second region 308 in which small air holes 309 are placed to form a higher effective index compared to that of the inner cladding region 306.Finally, the waveguide consist of an outermost cladding 310 in which air holes 311 of a medium size (compared to those of the inner regions) are placed.","It should be noted that we throughout this description generally will assume that the basis material is silica or doped silica, since this is the most commonly used material for fabrication of optical fibers.","However, the principles and ideas behind the invention is not limited to these material combinations, and it may for future applications, where new spectral ranges of the optical fiber technology is explored, be more advantageous to use different material compositions, such as (but not limited to) different compound glasses (e.g.","chalcogenide), polymers and/or low-melting point glasses.","Microstructured fibers are generally divided into two types depending on the physical mechanism responsible for the light guidance: index-guiding microstructured fibers are characterised by a core having a higher refractive index than a surrounding cladding region (as known from conventional optical fibers), and photonic band gap micro-structured fibres are characterised by a core having effectively a lower refractive index than a surrounding cladding region having periodically distributed features (see Broeng et al, Optical Fiber Technology, Vol.","5, pp.","305-330, 1999 for a presentation of the classification of the two groups).","While the physical mechanism causing the waveguidance in the two types of microstructured fibers is different, the improvements disclosed by the present inventors relate to both types of microstructured fibers.","In FIG.","4 (prior art) is depicted how the effective index of the fundamental space filling mode (the “effective refractive index”), β/k, of a periodic structure varies with the normalised wavelength, λ/Λ.","Here λ is the free space wavelength of light, Λ is the distance between the centres of two nearest neighbouring air holes, β is the propagation constant and k is the free space wave-number, all of which are well known by those skilled in the art.","The periodic structure consists of circular air holes (the refractive index of air is 1.0 in the calculations) placed on a triangular lattice on a silica background (the refractive index of silica has been approximated by a refractive index of 1.45).","Three curves are shown: the curve at the top 401 is for a periodic structure where the air holes are placed in a triangular lattice (also known as close-packed arrangement) and have a diameter, d, of 0.2Λ, where Λ is the lattice period (equal to the center-to-center of the air holes for the triangular lattice).","The curve 402 is for a similar structure but with d=0.3Λ, while the curve at the bottom 403 is for a similar structure but with d=0.5Λ.","Notice that the effective refractive index is low and not strongly dependent on the wavelength when the wavelength, λ, is large compared to the structural size (symbolised by Λ).","It is known from the prior art that a low refractive index of the cladding is an advantage if strong dispersion compensation is desired.","If a photonic crystal fiber (PCF) with a silica core region (see FIG.","1b), surrounded by a periodic cladding region (such as the ones for which effective indices have been calculated as shown in FIG.","4) is to be used as a dispersion compensating fiber one may, therefore, assume that an inner cladding region with a small pitch, compared to the wavelength, is to be used, since this gives the lowest cladding index according to the effective index curves for periodic structures shown in FIG.","4.Furthermore, since the effective index of the cladding according to FIG.","4 is lowest when large air filling fractions are employed, one may assume that one should employ cladding structures with large air-filling fractions to obtain maximum dispersion compensation.","This is in agreement with the teachings by Birks et al.","in IEEE Photonics Technology Letters, Vol.","11, pp.674-76, 1999 and Optical Fiber Communication Conference, FG2-1, pp.","108-10.1999, and DiGiovanni et al.","in U.S. Pat.","No.","5,802,236.In the Birks-references, the cladding index is assumed to be one (which is of course less than the actual index of any real life microstructured cladding), while the core is supposed to be a circular glass-rod of very small diameter (0.4-1.2 μm).","According to the invention, PCFs for dispersion compensating purposes can, therefore, be made with a cross-section similar to the one shown schematically in FIG.","1b.","If the air-filling fraction of the cladding is large, the group velocity dispersion will typically have minimum values when the inter-hole distance of the cladding is small compared to the wavelength.","This corresponds to a core diameter, which is small compared to the wavelength, in agreement with the findings by Birks et al.","One should notice that a number of problems are associated with small core sizes: coupling light into the fiber using standard splicing techniques is associated with significant coupling losses because of a poor mode-overlap with the standard fiber coupled from or to.","This is well understood by those skilled in the art.","Further, small core sizes enhance non-linear effects in the high bit rate systems of the future.","This is also well understood by those skilled in the art.","However, the group velocity dispersion values that may be obtained, using the schematic design shown in FIG.","1b, are very impressive: group velocity dispersion may be as low as −2000 ps/nm/km according to Birks et al.","It is a disadvantage of the microstructured fibers with a cross-section similar to the one depicted in FIG.","1b with group velocity dispersion less than −400 ps/nm/km, that the fibers have small core size of around or less than 1 μm and, therefore, poor coupling coefficient to standard optical fibers.","It is a further disadvantage of the microstructured fibers with a cross-section similar to the one depicted in FIG.","1b with group velocity dispersion less than −400 ps/nm/km, that the fibers have small effective mode-areas and, therefore, suffer from non-linear effects in the future high bit rate transmission system.","Such fibers would, therefore, have a detrimental effect on the bit rates that can be transmitted by the future multi-gigabit systems.","The present inventors have realised that a number of ways exist of designing microstructured fibers with large negative group velocity dispersion, while avoiding the small core sizes that results from the large index contrast inherent in the designs suggested so far for dispersion compensating PCFs.","To understand the basic idea, consider again the mode-indices of FIG.","4.If one could design a fiber with a core index that abruptly rises above the cladding index as the frequency is increased, one would obtain a mode-index that curves upward in an index plot similar to the index plot shown in FIG.","4.This corresponds to large negative group velocity dispersion, GVD, according to the formula for group velocity dispersion: GVD = - λ c ⅆ 2 n ⅆ λ 2 .","The only apparent problem with this approach is that the index curves shown in FIG.","4 never seem to cross.","However, the index curves shown in FIG.","4 are for a specific class of periodic structures.","The problem has already been solved for microstructured fibers of the prior art: By enlarging the inner cladding air-holes, such as shown schematically on FIG.","2b, the field is not allowed to be guided in the core region with a mode-index, which exceeds that of the cladding index at large wavelengths.","At some wavelength, the mode-index of the fundamental cladding mode rises very abruptly, leading to large negative group velocity dispersion over a limited wavelength range.","This is because the fundamental mode is localised within the central core region at short wavelengths, which causes the mode-index to rise abruptly, while the mode at the same time becomes guided.","Unfortunately, this abrupt rise in mode-index happens, when the core region is relatively small compared to the core region of standard optical fibers, thus giving the same problems with non-linear effects and poor coupling as the basic fiber design depicted in FIG.","1a.","The present inventors have realized, however, that by doping the central core region, giving it an index of refraction that is higher than the index of refraction of the background material of the cladding region, one may obtain large negative group velocity dispersion, while having larger core regions than the core regions of the prior art fibers.","Such a design is shown schematically in FIG.","5, which shows the higher refractive index of the core region 500, the inner cladding air holes 501, the outer cladding air holes 502 and the background material 503 with a lower refractive index than 500.It should be noticed that the doped core is not in contact with the inner cladding features 501.As shall be discussed in more details later, in connection with FIG.","31, the present inventors have realized that this provides an advantageous design direction for lowering the mode field overlap with the low-index features and thereby lowering potential loss-mechanisms associated with the low-index features and their interfaces with the background material of the fiber (such as absorption and scattering).","In FIG.","6 (prior art) there is illustrated a number of mode-indices of periodic structures, which illustrates why effective dispersion compensation may be obtained by a fiber design as illustrated in FIG.","5.The curve at the top 601 repeats the effective index of the curve 401 of FIG.","4—this curve is referred to as the outer cladding index, since it is supposed to represent the outer cladding structure of the micro-structured fiber shown in FIG.","5 (502 and 503).","The illustration also shows the effective indices of hypothetical honeycomb periodic structures, where the background has an index of refraction of 1.46.In a sense, these honeycomb structures may represent a periodic version of the central core region and its surrounding 6 holes of inner cladding structure—see for example Riishede et al.","Proc.","27th Eur.","Conf.","On Opt.","Comm (ECOC'01), pp.","Th.A.1.5, 2001.Three curves are shown: curve 602 corresponds to a honeycomb-structure with a hole diameter, d, of 50% of the inter-hole distance, Λ, (center-to-center), 603 corresponds to d=0.6Λ, and 604 corresponds to d=0.7Λ.","The present inventors have realized that any strictly guided modes must have an effective index that lies above the effective cladding index, yet below the effective refractive index of the core region (approximated by the honeycomb-index).","At some wavelength (λ/Λ around 0.5-0.6 for the design parameters of this example), the honeycomb indices are seen from FIG.","6 to rise above that of the outer cladding index.","This is caused by the higher refractive index of the central core region compared to the background refractive index of the cladding region.","The central core, therefore, only guides light at shorter wavelengths than the wavelength, where the honeycomb index and the cladding index cross (to the left of the crossing point in FIG.","6).","As before, the guiding is associated with the fundamental mode being localised to the core region, thus giving large negative group velocity dispersion, when the cladding index abruptly rises above the core index—here the mode field distribution of the fundamental mode will shift away from the central core.","However, by doping the central core we can employ small holes—or complete elimination of these—in the outer cladding structure, and quite large holes in the inner cladding region, and still obtain a crossing wavelength.","Actually, a crossing wavelength will always exist, if the core region has a region with higher refractive index than any index of refraction in the outer cladding region.","One may vary the hole-size and obtain different crossing points.","FIG.","6 also illustrates that enlarging the inner cladding air holes moves the crossing point to the left.","This is an advantage, since for a fixed operational wavelength, λ, this corresponds to a larger structure size, Λ, and, therefore, a larger core region.","To exemplify; 602 crosses 601 at λ/Λ=0.57, 603 crosses 601 at λ/Λ=0.46 and 604 crosses 601 at λ/Λ=0.44.Choosing λ=1.55 μm as the operation wavelength this corresponds to Λ=2.72 μm for 602, Λ=3.37 μm for 603 and Λ=3.52 μm for 604.One can, therefore, adjust the core size corresponding to large negative group velocity dispersion relatively freely, by simply adjusting the inner cladding hole size.","The above-mentioned Λ-values correspond to inscribed core diameters of between: 4.1 μm for 602, 4.7 μm for 603, and 4.6 μm for 604.Other possibilities of adjusting the core size to a desired value exist, however.","The cladding index 601 is repeated as 701, and the honeycomb index for a fiber with an inner cladding-hole diameter of 60% of the inter-hole distance 603 is repeated as 703 in FIG.","7.The diameter of the core region with refractive index of 1.46 is equal to the inter-hole distance in 703.In FIG.","7 is also shown the dramatic effect of changing the doping levels of the central core region: If the refractive index of the region with increased index is raised to 1.47 (curve 702) while maintaining the diameter (relative to the hole distance) of the doped region, the crossing point may move significantly to the right (from λ/Λ=0.46 to λ/Λ=0.72).","This can be counter-acted by reducing the diameter of the doped region to 50% of the inter-hole distance as shown by 704.For a wavelength of 1.55 μm (curve 702) corresponds to Λ=2.15 μm, curve 703 corresponds to Λ=3.37 μm and curve 704 corresponds to Λ=3.52 μm at their respective crossings with curve 701.A large number of possibilities, therefore, exist for adjusting the core size (and thereby the mode field diameter) with this design.","These possibilities include: 1) The value of the cladding index.","The higher the cladding index the larger the core for a given core design.","The present inventors have realised that largest core region is obtained when the outer cladding is a homogeneous material (such as pure, fused silica or silica glass doped with for example Fluorine).","2) The size of the inner cladding air holes.","3) The maximum of the refractive index in the doped core and the refractive index profile.","Generally, a larger refractive index step between the central core and the background material of the outer cladding leads to a smaller core size as shown by FIG.","7.4) The diameter of the up-doped central part of the core region.","Generally a smaller diameter leads to larger Λ, and thereby a larger core area, and larger mode field diameter, as shown by FIG.","7.The actual combination of parameters that should be employed depends on the specific application of the micro-structured fiber.","For dispersion and dispersion slope compensating applications, there is a general interest in core regions that are relatively large in order to eliminate degrading effects caused by non-linearity.","The present invention, therefore, applies to fibers with a core diameter that is more than three times the free space wavelength of the light that should be dispersion compensated.","For standard purposes (dispersion compensation in the so-called communication window) this means that that the core diameter of the fibers of the present invention should be larger than 4.0 μm.","On the other hand, the design ideas outlined in the present invention may also be used to tailor the dispersion to zero or close-to-zero at wavelength around 1550 nm for fibers with small core sizes—such as core sizes below 4 μm in diameter.","The present invention, therefore, also covers fibers for non-linear applications.","The present invention does not limit itself to structures with the principal design shown in FIG.","5.It is well known that dispersion-compensating fibers with a depressed cladding region tend to have large bending losses.","As a general rule, the bending losses increase when a larger core region is employed.","Since the present invention in a main aspect relates to large core regions it would be advantageous to design the fiber such that bending losses are suppressed.","The present inventors have realised that this is indeed possible with the type of design shown schematically in FIG.","8.The core region 800, the inner cladding region comprising low-index features 801, and the outer cladding region 803 and 804 are similar to those shown in FIG.","5.However, some of the innermost cladding air holes have been removed at positions 802.This effectively creates what may be termed a raised outer cladding region (similar to that shown in FIG.","3 and denoted 308), or alternatively it may be termed an outer core region 802 if one follows the line of thought presented by Thyagarajan et. al.","in IEEE Photonics Technology Letters, Vol.","8, no.","11, November 1996, pp.","1510-12.Note, however, that compared to the approach by Thyagarajan et al., the microstructured fiber does not need high doping levels, which are difficult to control, and we may even operate with much larger effective index contrasts.","Since one may in some sense view the inner core 800 and the inner cladding 801 as one core (which was theoretically exploited to let honeycomb indices represent the core index in FIG.","6), we approximately know the slope of the effective guided mode index of the central core region.","Looking at FIG.","6, we realise that strong negative group velocity dispersion will result, if the slope of this mode is numerically large, while the slope of a mode guided mainly within an outer core is numerically small.","In designs such as that shown in FIG.","3a, the negative group velocity dispersion depends critically on the type of design.","If 303 cannot be seen as capable of supporting guided mode(s), only modest dispersion may result (a typical result will be less than nominally 300 ps/nm/km).","If on the other hand 303 represents an outer core, strong dispersion may result (even more negative than −5000 ps/nm/km).","It should be understood that by using the term “represent an outer core” is meant that the core may support a guided mode, but not necessarily does so—for example if no light is coupled into the core from the input end of the fiber.","With designs such as that shown in FIG.","8 on the other hand, the numerical value of the dispersion may be large in both cases (and actually can be equally large in structures such as the one shown in FIG.","5).","This is because the core index may have a large negative slope, when it crosses the cladding index, as illustrated in FIG.","6.One additional functionality is that the numerical value of the group velocity dispersion of micro-structured fibers such as shown in FIG.","8, therefore, does not depend critically on whether the outer core guides an independent mode in region 802 or not—it may still affect the guided mode in the central core 800.It may still be advantageous to have an outer core capable of supporting a mode.","This is because this will lift the effective index of the fundamental mode above the effective cladding index at the wavelength regime of strong dispersion and thereby reduces the bending losses.","FIG.","9 shows the operation of a microstructured fiber with the design schematically shown in FIG.","8.Curve 901 shows the effective core index as a function of the normalised wavelength, curve 903 shows the effective cladding index as a function of the normalised wavelength and curve 902 shows the effective mode-index of the fundamental mode as a function of the normalised wavelength.","For this fiber, the refractive index of the up-doped central part of the inner core 800 is 1.46.The diameter of 800 is equal to the inter-hole distance, Λ.","The refractive index of the background material of the rest of the fiber 804 is 1.45.The diameter of the inner cladding air holes is 0.6Λ.","The diameter of the outer cladding air holes is 0.2Λ.","Six of the innermost outer cladding holes were removed to create the outer core region 802 in the manner shown in FIG.","8.Looking at FIG.","9, it is seen how the mode-index of the fundamental mode 902 has a slope similar to the mode-index of the cladding 903 at long normalised wavelengths.","At normalised wavelengths near 0.45 the slope of the fundamental mode 902 changes and approaches the slope of the core index 901.This gives a strong upward curvature on 902 that correspond to numerically large negative group velocity dispersion of the fundamental mode 902.In this wavelength regime, where the fiber exhibits strong negative dispersion, the central and the outer core may support modes of approximately similar effective index.","The actual group velocity dispersion is shown in FIG.","10 for a fiber such as the one described above for FIG.","9.FIG.","10 is for a fiber with Λ=3.45 μm.","This corresponds to a central core 800 with refractive index=1.46 and diameter=3.45 μm.","The inner cladding air holes have diameter 2.07 μm (giving an inscribed diameter of 4.8 μm), while the outer cladding air holes have diameter 0.69 μm.","The background refractive index is 1.45.In agreement with the effective index variation 902, FIG.","10 shows strong negative group velocity dispersion, GVD, of less than −1600 ps/nm/km.","Designs with larger doping levels show even numerically larger group velocity dispersion values (down to −10000 ps/nm/km).","Also the large negative slope on the group velocity dispersion at wavelengths shorter than 1.55 μm should be noticed.","Such a large dispersion slope makes these fibers ideal for dispersion slope compensation schemes.","In some instances, the slope of these dispersion compensation fibers may even be too large.","However, systems having smaller slope (relative to the GVD-values) may be created by cascading fibers with different GVD characteristics.","It is, therefore, an advantage of these fibers that they may be used to not only compensate the dispersion but also the dispersion slope of virtually any system with positive group velocity dispersion.","In order to illustrate the good mode confinement of the dispersion compensating fiber designs according to the present invention, FIG.","11 shows the mode field distribution of the fundamental mode calculated at a wavelength of 1550 nm.","The fiber parameters are selected according to the description provided in connection with the illustration of the group velocity dispersion in FIG.","10.The field is seen to avoid the air holes in the inner cladding region, being advantageous in terms of low losses—as previously discussed.","This being mainly attributed to the core region having a high-index features being not in contact with the inner-cladding region.","In a preferred embodiment of the present invention, the outer core contains material with a higher refractive index than the refractive index of the background material in the outer cladding.","An example of a fiber that has a number of high-index features present in the outer core is illustrated in FIG.","12.In order to reduce fabrication efforts, it may further be preferred to eliminate the use of low-index features in the outer cladding.","The fiber in FIG.","12, therefore, consists of a first high-index type core region 1200, a single layer of low-index features 1201 surrounding the core and a single layer of high-index features 1202 surrounding both the core region 1200 and the low-index features 1201.Outside the core region only a single background material 1203 is present—this background material being similar for the inner cladding region, the outer core region and the outer cladding region.","Due to the higher refractive index of the features 1202 compared to the background material and the low-index features 1201, the ensemble of high-index features may act as a core in its own right that may support a propagating mode.","The outer core may also be formed from an annular region 1204 of high refractive index material.","The two core regions (these being the central core region 1200 and the ensemble of high-index features 1202 or the annular region 1204) thereby are able to support each their different core modes.","These core modes will be decoupled at most wavelengths and only couple to each other over a relatively short wavelength range.","Within this short wavelength range, the mode distributions in the fiber will change dramatically and the fiber will exhibit a strong dispersion.","The qualitative operation of this type of fiber is schematically illustrated in FIG.","13.As a function of wavelength, FIG.","13a shows schematically, the effective index of a mode that is guided in a fiber comprising only the central core region 1200 and of a mode that is guided in a fiber comprising only the outer core 1201 (this outer core mode being supported by the ensemble of high-index features 1203 or the annular region 1204).","The two mode curves are seen to cross at a normalised wavelength, λcoupling/Λ.","Around this wavelength, the two cores will couple strongly to each other for a composite fiber comprising both cores.","At the coupling wavelength, for a fiber with a design as for example shown in FIG.","12, the effective indices of modes supported by the two cores will have a lifted degeneracy—or so-called avoided crossing will take place.","In this manner the fiber is capable of guiding light in a fundamental mode corresponding to the curve 1303 that exhibits a strong negative dispersion at wavelengths around λcoupling/Λ.","The fiber may support a second mode guided indicated by the curve 1304, but it is not necessary that the fiber carries any substantial energy in the mode 1304 in order to exhibit negative dispersion in mode 1303.The fiber of FIG.","13b will guide light in the central core at wavelengths shorter than the wavelength of strongest coupling (roughly equal to the coupling wavelength indicated in FIG.","13) and the fiber will here exhibit negative dispersion with a negative dispersion slope—this is the range that dispersion compensating fibers typically are preferred to operate in.","The wavelength-axis in FIG.","13 has been normalised with respect to pitch (for example the center-to-center spacing between two adjacent low-index features 1201) to indicate that the coupling wavelength may be changed by changing the pitch.","The figure also indicates the index of a homogeneous cladding with a refractive index of around 1.444—corresponding to pure silica being used as background material 1203.The higher refractive index of the core 1200 as well as of the high-index features 1202 compared to the background material 1203 cause the coupling wavelength to occur in a regime where both the index of both core modes are above the cladding index.","In this manner, the coupling—and thereby the strong dispersion—may be obtain while the fiber is not sensitive to bending losses.","It is worth noticing that the mode indices of both core modes may be lower than the cladding index at long wavelengths.","This is due to the low-index features 1201 having a refractive index significantly below that of the cladding.","The core modes may, however, well be guided for mode indices below the cladding index—as is the case for conventional one-material photonic crystal fibers—but the modes may in this case experience leakage losses.","These leakage losses are typically reduced for one material photonic crystal fibers by having a sufficient number of low-index features surrounding the core region that supports a propagating mode—and the outer cladding may be micro-structured as for the fiber in FIG.","8.For homogeneous outer cladding regions, the potential problem of leakage-loss at the coupling wavelength may be eliminated by the use of features 1202 having a higher refractive index than the background material 1203.It is an advantage that there are different maximum refractive indices in the two core regions—and that the core region with the lower maximum refractive index has a larger special extend than the other core region.","As an example, the inner or central core region (referred to as first core) may have a maximum refractive index of 1.47 and the outer core region (referred to as second core) may have a maximum refractive index of 1.465.The first core should then be designed to have the smallest cross-sectional area of the two cores, whereby the index curve of the mode guided in the first core will have a more steep slope than the index curve for the mode supported by the second core.","The magnitude of the negative group velocity dispersion depends on various things.","The strength of the coupling between the two modes may be quantified by the field overlap.","At λcoupling/Λ (where the two un-coupled modes cross) the splitting of the two modes will be proportional to the coupling strength.","Often the coupling strength will only have weak wavelength dependence and in that case the wavelength-range where coupling is efficient in changing the modes is strongly related to the slopes of the two mode curves.","When the slopes of the coupling mode curves are as different as possible, the effective wavelength-range will be small and the modes (which have a curved behaviour) will get the smallest possible bending curvature, i.e.","numerically large group velocity dispersion will be exhibited when the slopes are as different as possible.","The present inventors have realized that microstructured fibers with a coupled-core design may also be advantageous for non-linear applications, where it is desired to have fibers with small core and a dispersion close to zero over a broad wavelength range (e.g.","around 1550 nm)—such as in the wavelength range from 1300 nm to 1700 nm (this is still in the regime of negative waveguide dispersion).","Looking at FIG.","13, it is seen that it will be preferred to move the coupling wavelength to smaller values of wavelength/pitch in order to reduce the fiber dimensions at a given wavelength.","Hence, it may be preferred that the two core regions have a maximum refractive index that is close to each other and that the area of the two core regions is not too different.","Alternatively, it is preferred that the outer core region has an effective refractive index close to the (effective) refractive index of the cladding.","According to the present invention, a number of characteristics are preferred for fibers as schematically shown in FIG.","12: Core region 1200 having a maximum refractive index, Nco,1.Typically, silica doped with Germanium and/or other doping materials will be used to form the core.","Various refractive index profiles of the core may be preferred, but typically a profile as close to a step-index profile will be preferred.","Hence, typically Nco,1 is larger than 1.444, such as preferably larger than 1.46, such as larger than 1.47 at wavelengths around 1550 nm.","Background material 1204 having a refractive index, Ncl,2, that is lower than Nco,1.Typically, the background material is pure silica, having a refractive index of around 1.444 at wavelength around 1550 nm, but silica doped with a doping material to lower the refractive index of Ncl,1 below 1.444 may also be preferred, such as fluorine-doped silica glass.","Low-index features 1201 having a refractive index, Ncl,1, a diameter, dcl,1, and a center-to-center spacing between two adjacent low-index features, Λcl,1.Typically, these low-index features are voids, hence Ncl,1 is around 1.0, but low index features formed from down-doped silica may also be preferred.","For dispersion compensation applications, typically the low-index features will have a diameter, dcl,1, in the range from around 0.5Λcl,1 to 0.9Λcl,1 and for non-linear fiber applications, typically, dcl,1 will be in the range from 0.2Λcl,1 to 0.5Λcl,1.In order to reduce degrading effects, such as polarisation mode dispersion, PMD, it is often preferred that all low-index features are identical and equally spaced.","Also other numbers of low-index features than 6 is covered by the present invention as well as more than one “ring” of low-index features surrounding the core region 1200.High-index features 1202 having a refractive index, Nco,2, a diameter, dco,2, and a center-to-center spacing between two adjacent high-index features, Λco,2.Typically, these high-index features comprise doped silica glass, hence Nco,2 is larger than 1.444 at wavelengths around 1550 nm, but high-index features formed from pure or down-doped silica may also be preferred.","For dispersion compensation applications, typically the high-index features will have Nco,2 in the range from 1.444 to 1.460 at wavelengths around 1550 nm and a diameter, dco,2, in the range from around 0.1Λco,2 to 0.9Λco,2 and for non-linear fiber applications, typically, dco,2 will be in the same range.","Furthermore, in order for the ensemble of high-index features to support only a single-mode, it is often preferred that the size of the high-index features is smaller than the size of the low-index features 1201.The present inventors have further found that it is an advantage that the refractive index profile of the high-index features is as close as possible to a step-index profile in order for the coupling to be as dramatic as possible—and thereby the dispersion as strong as possible.","In order to reduce degrading effects, such as polarisation mode dispersion, PMD, it is often preferred that all high-index features are identical and equally spaced.","Alternatively, the ensemble of high-index features in the outer core may be substituted by a homogeneous high-index annular region 1204.Such an annular region—being either the complete or parts of the outer core region—may have the same functionality as the ensemble of high-index features.","The width, the maximum refractive index and the index profile of the annular region are key parameters for the fiber.","Also other numbers of high-index features than 6 is covered by the present invention as well as more than one “ring” of low-index features surrounding the core region 1200.In order to optimise the coupling between modes supported by the first core region 1200 and the second core region (formed from the high-index features 1202), it is often preferred that the high-index features are positioned such that light penetrating between two low-index features from the core region 1200 will experience the high-index features most strongly.","To quantify this, it may be preferred that a first polygon 1400 drawn from centres of high-index features and a second polygon 1401 drawn from centres of low-index features are concentric, but rotated with respect to each other an angle given by 0.5(N1/360′), where N1 is the number of low-index features.","Hence, for a fiber with 6 low-index features (as in FIG.","14), the first polygon 1400 is rotated 30 degrees compared to the second polygon 1401, and both polygons are coinciding.","Alternatively, the second polygon may have a similar orientation as the first polygon.","As schematically illustrated in FIG.","15, it may also be preferred that the outer cladding region 1501 comprises a different material—or a material with a different refractive index—than the background material 1500 of the inner cladding and/or the outer core.","It is important to notice that the above-described design ideas for providing microstructured optical fibers for dispersion and dispersion slope compensation as well as for non-linear applications is profoundly different from the ideas presented by DiGiovanni et al.","in U.S. Pat.","No.","5,802,236, where a second core region capable of supporting a propagating mode is taught to be disadvantageous.","To illustrate in more detail the operation and some of the possibilities of fibers according to the present invention, various design parameters of fibers as shown in FIG.","12a shall be considered next.","FIG.","16 shows a simulation of a fiber according to the present invention having a design as schematically shown in FIG.","12.The fiber has the following design parameters: The first core has a step-index profile with a maximum refractive index of 1.471 (corresponding to a refractive index change of 1.892% compared to pure silica having refractive index of 1.444 at λ=1550 nm) and a background material of refractive index 1.444 (pure silica).","The inner cladding region comprises a background material of refractive index 1.444 and six voids of diameter 0.5Λ, where Λ is the center-to-centre spacing between to two nearest voids.","The second core region comprises a background material of refractive index 1.444 and six high-index features of refractive index 1.459 (corresponding to a raised index of 1.055% compared to pure silica).","The outer cladding region is homogeneous and comprises pure silica with a refractive index of 1.444.While the outer cladding region is homogeneous in this case, it may also comprise varies sub-regions for example for lowering bending losses.","As seen from FIG.","16, the fiber supports a fundamental mode 1600 that has a strongly dispersive behaviour at normalized wavelengths, λ/Λ, of around 0.36.The curve 1601 represent the effective index of a mode that may be supported by the second core and the curve 1602 represent the refractive index of the outer cladding region.","It should be noted that it is here chosen to use a normalisation factor, Λ.","The fibre in FIG.","16 will have Λ of around 4.3 μm for negative dispersion operation of around λ=1550 nm.","This provides a core size that is significantly larger than the core sizes disclosed by the previously discussed DiGiovanni et al.","and Birks et al.","references.","Furthermore, as shall be demonstrated next, the negative dispersion that may be exhibited by fibers according to the present invention can be significantly larger than for the fibers disclosed in the above-mentioned DiGiovanni and Birks references.","For tailoring fibers according to the present invention for given specifications, a wide range of parameters may be tuned.","The following figures show examples of how various fiber characteristics (such as dispersion, RDS and core size) may be tuned by adjusting design parameters.","The present inventors have found that the negative dispersion depends strongly on the size of the inner cladding features.","FIG.","17 shows a simulation of the dispersion characteristics of fibers as in FIG.","16, but with A ranging from 4.08 μm to 4.46 μm, and inner features being voids with a size that ranges from d/Λ of around 0.30 to 0.65.As seen from the figure, the dispersion has a characteristic “dip” with a dispersion minimum (the strongest dispersion) and a certain width.","It is worth noticing varying the size of the inner cladding features has a strong influence on the minimum dispersion values as well as the width of the “dip” whereas the wavelength of strongest negative dispersion is only varied slightly.","As seen from the figure, negative dispersion that is significantly beyond prior art dispersion compensating fibers is feasible.","FIG.","18 shows the normalized wavelength of strongest dispersion, λ*/Λ, as a function of size of the inner cladding features (solid curve).","The figure shows that by adjusting the features size, d/Λ, from around 0.25 to 0.70, λ*/Λ can be varied from around 0.34 to 0.385.For the strongest dispersion to occur at a wavelength of around 1550 nm, this corresponds to Λ varying from around 4.03 μm to 4.50 μm (dashed curve), and d ranging from around 1.0 μm to 3.2 μm.","These Λ values provide a relatively large separation between the high-index feature in the core 1200 and the air holes 1201.The distance from the center of the fiber to a center of an air hole 1201 is similar to Λ, hence, the afore-mentioned separation is given by s1=Λ−d−dco,1.With d/Λ ranging from 0.25 to 0.70 and dco,1 ranging from 0.5Λ to 0.8Λ this provides a separation s1 ranging up to 1.2 μm.","For other designs that are within the present invention, the separation may be larger as may be deduced from the various examples of fibers according to the present invention.","A non-zero separation, s1, provides an important technical advantage in terms of lowering potential losses associated with the air holes—as shall be discussed in further detail at a later stage.","Another important feature of fibers according to the present invention is the relatively large separation between the first core 1200 and the second core (ensemble of high-index features 1202 or ring-shaped element 1204).","This separation is typically larger than Λ, such as larger than 4.0 μm as apparent from the various examples of the present application.","Alternatively, this advantage is apparent in the relatively large diameter of the second core (the diameter being defined as the radial distance in the cross-section from the of the fiber to a center of the second core region (such as for example a center of one of the second core features 802, 1202 or a center of the ring-shaped region 1204).","As apparent from the various examples of fibers according to the present invention, e.g.","as illustrated in the various embodiments in the figures, this diameter is typically in the range from 10 μm to 20 μm.","Larger diameters may be desired for fibers operating in a higher-order mode—as shall be discussed at a later stage.","It is important to notice, that the above-described separations and diameters are significantly larger than for fibers disclosed by Fajardo et al.","in WO 01/98819.The above-described feature is advantageous in separating the two cores for more simple in-coupling (including splicing) such that light may be coupled substantially only into the first core region.","For splicing to a standard transmission fiber (which dispersion is desired to be compensated) it is an advantage that the second core has a diameter larger than the core diameter of the transmission fiber.","Typical transmission fibers have a core diameter of around 8 μm, and in order to avoid coupling into the second core at a fiber splice, it is an advantage that the diamater of the second core is larger than 8 μm (preferably larger than 10 μm) such that its overlap with the core of the transmission fiber is limited or negligible when the center of the first core of the dual core fiber is aligned to the center of the core of the transmission fiber.","Furthermore, in order to lower in-coupling losses, it is desired that the first core—and importantly the high-index feature of the first core—has a size that is approaching the size of the core of the transmission fiber nor an intermediate fiber that has been inserted between the transmission fiber and the dispersion compensating fiber to provide mode adaption.","In the prior art, intermediate fibers having a core size smaller than the transmission fiber is often inserted between the transmission fiber and the dispersion compensating fiber in order to adapt the core sizes.","Typically, intermediate fibers may adapt cores from around 8-9 μm in diameter to cores of down to 4 μm.","Hence, it is desirable to realize dual core dispersion compensating fibers with a first core having a diameter of around 4 μm to 8 μm and a second core diameter of larger than 9 μm.","As apparent from the present application, such a feature is characteristic for a number of fibers according to the present application.","The relatively large dimensions for the here-disclosed fibers provide several important technical advantageous as discussed throughout the application apart from the facilitated in-coupling—including relatively large MFD for lowering non-linear effects, lower PMD as lower birefringence is typically found for micro-structured fibers of larger dimensions and symmetric feature arrangements, lower losses as larger dimensions provide decreased field intensity in the air holes of the first cladding region for light guided in the first core.","One or more of these technical advantages may be obtained for fibers according to the present invention at the same time as a stronger dispersion is obtained.","FIG.","19 shows a simulation of the value of minimum dispersion (strongest negative dispersion) for a fiber according to the present invention having inner cladding features varying in size from d/Λ of around 0.25 to 0.70 (normalized dispersion is indicated with a solid line—right axis).","For operation around 1550 nm, it is possible to vary the dispersion from around −400 ps/nm/km to more than −10000 ps/nm/km (dashed curve—left axis).","As previously mentioned, such dispersion characteristics are significantly stronger than for any prior art optical fiber.","As seen from FIG.","17, the width of the negative dispersion “dip” depends strongly on the size of the inner cladding features.","Generally, a larger inner features size results in stronger dispersion, but a more narrow “dip”.","In FIG.","20, the width, λ, of the dispersion “dip” is simulated as a function of inner cladding features size (normalized width, □λ/Λ, is shown by solid line (right axis) and for operation at λ=1550 nm, the absolute width, □□, is shown by the dashed line (left axis)).","The width is determined as the full-width-half-maximum (FWHM) of the numerical value of the dispersion.","As seen from the figure, for a FWHM value of more, than 30 nm, the fiber should be designed with feature size, d/Λ, of around 0.45 or less.","FIG.","21 shows a calculation of the RDS values and the dispersion of another fiber according to the present invention having d/Λ=0.4 and Λ=4.214 μm.","As seen from the figure, the fiber exhibits a strong negative dispersion of more than −500 ps/nm/km as well as a high RDS value of around 0.03 nm−1 at a wavelength of 1550 nm.","Such a fiber is well suited for dispersion and dispersion slope compensation.","FIGS.","22 and 23 shows similar results for fibers having d/Λ=0.5 and d/Λ=0.6.FIG.","24 shows another example of a fiber according to the present invention.","In this figures, the outer core is a homogeneous material.","The figure shows the operation of the fiber, which is similar to the previously discussed fibers of FIG.","12.It is important to notice that the fibre exhibits an abrupt cut-off both of the fundamental mode 2400 at λ/a of around 0.28, as well as of higher-order modes at λ/a around 0.1, where a is the radius of the high-index feature in the core (the first higher-order mode is shown by the line 2403).","This behaviour is also characteristic for the other types of fibers disclosed in the present application.","The higher-order mode with a cut-off of around λ/a=0.1 corresponds to an LP11 mode that is supported by the first core.","This mode will exhibit a strong negative dispersion around λ/a=0.1 that may be utilized for dispersion compensating applications.","Hence, for operation at a given wavelength, the structural dimensions of the fiber should be scale approximately 0.28/0.10=2.8 times larger for operation in the higher order mode.","An operation around λ/a of 0.1 corresponds to a value of around 15 μm for operation at a wavelength of around 1550 nm.","This results in a significantly larger core size compared to the core size of the previously discussed fibers operating in a fundamental mode.","Furthermore, as the cut-off of the higher-order mode is seen to be even more abrupt for the higher order mode compared to the fundamental mode—even stronger negative dispersion may be obtained.","For the single-mode dispersion compensating fiber, λ=1.55 μm, dictates an a-value of around 5.5 μm, and for the higher-order mode dispersion compensating fiber, an a-value of around 15 μm is dictated.","The present invention also relates to higher-order mode dispersion compensating fibers for other fiber designs that are disclosed by the present invention—dimensions must naturally be scaled and relations between design parameter may changes for transfer of a design for single-mode dispersion compensation to higher-order mode dispersion compensation.","FIG.","25 shows another schematic example of a dispersion compensating fibre according to the present invention.","The fiber is characterized by a first core region 2500 in the center of the fiber, an inner cladding region (a first cladding region) comprising low-index features 2501 and a background material 2502, a second core region 2503 and an outer cladding region 2504 (a second cladding region).","The first core 2500 has a size being adapted such that the core makes contact with features in the inner cladding 2501—this type of core has also been discussed by Fajardo et al.","in WO 01/98819.The first core 2500 is characterized by a refractive index being higher than the refractive index of the features 2501 as well as that of the background material 2502 of the inner cladding.","The second core 2503 that may either be homogenous (as shown in FIG.","25) or it may contain a number of micro-structured features (as discussed previously for example in FIG.","13).","The second core comprises material with a higher refractive index compared to the effective index of the inner cladding region and a lower refractive index compared to a maximum refractive index of the first core.","In a preferred, embodiment, the background material of the inner cladding region 2502 is identical to the refractive index of the second core 2503.The material of 2502 and 2503 may in this case be identical such as pure silica or silica comprising one or more index-raising dopants, such as for example Ge.","An important issue for dispersion compensating fibers is their bending losses.","The present inventors have analysed a number of fibers according to the present invention and found an important design parameter related to the cross-sectional size of the second core (or the total area of high-index feature in the second core in the case of an ensemble of high-index features forming the second core).","This parameter is important for lowering bending losses of the fibers as shall be demonstrated using the FIGS.","26 and 27.Looking first at FIG.","26, the fibre is characterized by a first core with a refractive index of 1.461 and the inner cladding region comprises six air holes with diameter, d. The background material of the inner cladding region is similar to the material of the outer core and it has a refractive index of 1.444 (pure silica).","The outer cladding comprises silica with a refractive index of 1.437, this may be obtain for example by F-doping of silica.","The fiber in FIG.","26 has an outer radius of the inner cladding region labelled R and an outer radius of the second core labelled R1.The fiber has R1/R=2 and d/R=0.37.In FIG.","26 is illustrated a simulation of the operation of the fiber.","The figure illustrates the effective indices of various modes of the fiber as a function of normalized wavelength, λ/2R.","The curve 2600 represents the fundamental mode guided in the core region, the curve 2601 represent a higher order mode that is supported by the fiber, but not necessarily carries any energy, and the curve 2602 represent the refractive index of the outer cladding region.","The figure shows that the curve 2600 has a strongly dispersive behaviour at normalized wavelengths around 0.9.The dispersive behaviour is noticeable by the “bending” of the curve 2600.This shows that the fiber exhibits a strong negative dispersion at normalized wavelengths around 0.9—and the fiber may, therefore, in principle, be used for dispersion compensating applications.","There is, however, an important issue to consider for this fibre.","This issue relates to the index difference between the curves 2600 and 2602.The two curves have a relatively small difference in the wavelength regime of strongly dispersive behaviour (for this specific example the difference is around 0.001 or less at the wavelength of strongest dispersion).","This small difference has the negative effect that the mode corresponding to curve 2600 will not be strongly confined at the strongly dispersive regime and may experience high bending losses.","The present inventors have realized that the potential problem of bending losses may be solved by careful design of the second core—more specifically by increasing the cross-sectional area of this region—or equivalently its width.","As shown in FIG.","27, the present inventors have found that for R1/R above 2, such as around 4, the strongly dispersive behaviour of the curve 2700 (to be compared with 2600 in FIG.","26), may be lifted significantly above the refractive index of the outer cladding 2702—thereby lowering the bending losses of the fiber.","Hence, it is advantageous to have the outer diameter of the second core cladding region being more than two times larger than outer diameter of the inner cladding region (this diameter here being defined as 2R), and preferably around 4 times larger.","Naturally, as those skilled in the art will understand, the exact position of the strongly dispersive regime depends on a number of design parameters including the refractive index of the various materials that the fiber is composed of as well as the morphology of the various microstructures and regions in the fiber (among others, these being size, shape and arrangement).","Considering next the dispersion that may be obtained for the fiber of FIG.","27b, FIG.","28 shows the normalized dispersion (solid curve), where the factor 2R has been used for normalization.","FIG.","28 also shows the effective mode area (dashed curve) of the fundamental mode.","As seen, the effective area is strongly increasing as the negative dispersion regime is reached (it should be noticed that the fiber should be operated at the short wavelength side of the dispersion dip).","The large increase in effective area is attributed to the fundamental mode being coupled to the second core at wavelengths longer than the minimum dispersion wavelength.","It is at this stage worth comparing the fiber in FIG.","28, to one of the previously studied fibers with a core not being in contact with the inner cladding features.","FIG.","29a shows dispersion and effective area of the fundamental mode for a fiber with a design as shown in FIG.","12a.","In FIG.","29a, a normalization factor Λ (center-to-center hole spacing in the inner cladding region) has been employed.","For a more direct comparison using absolute values of the design parameters, the two fibers of FIGS.","28b and 29b may be designed for dispersion compensation around λ=1550 nm.","FIG.","30 shows the effective area of the fundamental mode of the fiber of FIG.","28 (dashed curve 3001) and of FIG.","29b (solid curve 3000).","As apparent from the figure, both fibers may exhibit an effective area of more than 15 μm2 at wavelengths around 1550 nm, and in fact more than 20 μm2 may be exhibited by both fibers.","While this is comparable to conventional dispersion compensating fibers, it is significantly larger than for the fibers disclosed in the previously discussed DiGiovanni et al.","and Birks et al.","references.","Considering next the dispersion of the fibers according to the present invention, FIG.","31, shows that both fibers may exhibit a negative dispersion around 1550 nm that is significantly larger than what is obtained using conventional dispersion compensation fibres as well as the fibers disclosed by Hasegawa et al.","in EP 1 118 887.Hence, the present invention provides new dispersion compensating fibers have effective areas being comparable to conventional dispersion compensating fibers, but with significantly larger negative dispersion.","For a further comparison of fibers having a high-index core in direct contact with the inner cladding features and a high-index core being separated from the inner cladding, FIG.","31 shows that a stronger dispersion may be obtained in the latter case.","A further important issue is the mode field distribution in and around the core at the wavelengths of negative dispersion.","Looking at FIGS.","28a and 29a, it is found that the negative dispersion occurs for λ/2R of around 0.8 for the fiber in FIG.","28b and for λ/Λof around 0.36 for the fiber in FIG.","29.For operation of around 1550 nm, this yields 2R of around 1.9 μm and Λ of around 4.3 μm.","Hence, the inner cladding features for the fiber in FIG.","29b will be positioned substantially further away from the fiber center compared to the inner cladding features of the fiber in FIG.","28.Since both fibers have approximately similar effective area at wavelengths around 1550 nm (as shown in FIG.","30), the field overlap of the guided mode with the inner cladding features will be substantially lower for the fiber in FIG.","29b compared to the fiber in FIG.","28.Generally, the inner cladding features are air holes and may be desirable to limit the amount of field in the air holes in order to lower potential loss mechanism attributed to the air holes (this may for example be absorption losses from OH content in the air holes or scattering losses from surface roughness of the air holes).","Also in the case of lowering PMD, it may be an advantage to move the air holes as far away from the mode field as possible, since the high index contrast between the air holes and the background material may result in significant polarization effects.","For these reasons it may be preferred to have dispersion compensating fibers according to the present invention where the first core comprises a high-index features being isolated from the inner cladding features.","Preferably, the first core feature has a diameter of around 0.7Λ or less.","In other cases, however, it may be preferred to have as large an overlap between the mode field and the inner cladding features as possible.","This may for example be the case for tuneable dispersion compensating fibers, where the material in the inner cladding features may be altered in one way or another (for example by changing the refractive index of the inner cladding features dynamically using non-linear optical, acoustic-optical, or thermal processes, other mechanisms could be physical pressurizing or bending of the fibers).","This may be particularly interesting for dispersion compensating fibres according to the present invention comprising polymers in the inner cladding features.","Numerous possibilities exist for obtaining the required dispersion compensating effect as described in the previous text.","In the following a number of further examples of fibers according to the present invention shall be presented.","In FIG.","32 is schematically shown a preferred embodiment of the desired fiber.","Here the cross-section of a micro-structured fibre for dispersion compensation is shown having a doped central core 3201 surrounded by an annular inner cladding section containing more closely placed air holes 3202 than those used to form the micro-structured outer cladding 3203.Between the inner and outer cladding regions, a higher effective index is obtained in an annular segment 3204, where air-holes have been removed (compared to the air-filling fraction of the outer cladding).","It should be noted that the closer spacing of the holes provides the desired lowering of the effective index, but the present type of design becomes more simple from a manufacturing point of view, because it indicates how similar effects may be obtained both through control of air-hole spacing and through control of their size.","Yet another example of a preferred embodiment of a fiber according to the invention is schematically shown in FIG.","33.In this embodiment, the cross-section of a micro-structured fibre for dispersion compensation is realized by having a doped central core 3301 surrounded by an annular inner cladding section containing more closely placed air holes 3302 than those used to form the micro-structured outer cladding 3303.This is in accordance with the example illustrated in FIG.","32.However, compared to previous examples, additional air holes have been removed in the outer core region 3303, hereby allowing for a higher effective refractive index.","Comparing FIG.","32 and FIG.","33, we may formulate the differences as follows.","The fibre structure presented in FIG.","32 has an outer annular core section formed by a micro-structured section (in which the air-hole spacing is larger than that of the outer cladding), whereas the structure illustrated in FIG.","33 has an outer core region formed by a solid material.","It should be noted that these examples only indicate some of the design possibilities and optimal designs according to the parameters specified in this invention include the possibilities that not only size and spacing of air holes but also background doping may be used for example using high-index features or a annular element having a center that coincides with the fiber center—as previously discussed.","The outer core region may for example contain a number of solid, high-index features 3400 as illustrated in FIG.","34—or be a homogeneous annular region 3500 with refractive index higher than the refractive index of the outer cladding background material, as illustrated in FIG.","35.In the examples of the invention presented so far, the microstructured outer cladding has been illustrated as close packed air holes placed in a background material.","It should be stressed that other air-hole distributions may be used to obtain the desired optical properties.","Examples of such placements of air holes could be in rectangular-, honeycomb-, or Kagomé structures.","However, even further possibilities exist, and one preferred embodiment is schematically shown in FIG.","36, where the cross-section of a microstructured fiber according to the invention is shown.","In this preferred embodiment the outer cladding is formed by air-holes 3604 placed in concentric circles.","The fiber has a doped central core 3601 surrounded by an annular inner cladding section containing concentrically placed air holes 3602.Between the inner and outer claddings, the second core region 3603 is shown.","Note that in this specific example, the second core region is formed by a homogeneous material (such as pure silica or silica doped with germanium, fluorine and/or other material(s)), but it could also be realized through a microstructured section containing low- and/or high-index features.","The air holes that are used to tailor the effective refractive index do not necessarily have to be of circular cross sections.","In FIG.","37, a preferred embodiment of a fiber according to the invention is schematically shown.","FIG.","37a illustrates the cross-section of a micro-structured fibre according to the invention, in which the outer cladding is formed by air-holes 3704 placed in concentric circles, and where the inner cladding is formed by air holes of a non-circular cross section 3702.As in the previously discussed embodiments of the invention, the fiber has a doped central core 3701 surrounded by the annular inner cladding section.","Between the inner and outer claddings, the second core region 3703 is shown.","FIG.","37b shows yet another preferred embodiment having a doped central core 3705 surrounded by an annular inner cladding section containing non-circular air holes 3706.Between the inner cladding and the outer cladding 3708, the second core region 3707 is shown.","To further illustrate this difference between standard fibers and micro-structured fibers, we move on to showing the possibility of having a mode with large negative dispersion in a design where one would expect this to be impossible using standard index analogies between micro-structured fibers and standard fibers, such as those shown in FIGS.","1-3.We employ a central core design such as the one shown in FIG.","8, however, we assume the doping material does create a material with a refractive index being lower than the refractive index of silica (one could dope with e.g.","fluorine to obtain this effect).","Surrounding the inner cladding is an outer silica core, while the outer cladding is a micro-structured cladding with a small pitch.","According to the standard view of the core regions (such as illustrated in FIGS.","1-3) we, therefore, have an inner core with a refractive index being lower than the refractive index of the outer core.","According to this view, the fundamental and the second order mode will, therefore, swap place (the fundamental mode will be in the outer core at short wavelengths, and in the inner core at long wavelengths) compared to the design suggested by Thyagarajan et al.","in IEEE Photonics Technology Letters, Vol.","8, pp.","1510-12, 1996.This has detrimental effects on the performance of the fiber, which is the reason why Thyagarajan et al.","suggested the higher index in the central core.","The present inventors have, however, realized that it may also be preferred that the central core region does not comprise the highest refractive index material of the fiber.","FIG.","38 illustrates the effective core index 3802 (which is the honeycomb-index corresponding to the inner core and the inner cladding) and the cladding index 3801.The core index 3802 is seen to cross the cladding index 3801 while 3802 has a large negative slope.","This will make it possible to have a fundamental mode with the desired dispersion characteristics, provided that the silica regions of the outer core are sufficiently small to ensure that the effective mode index of the outer core is near the outer cladding effective refractive index.","Notice, that this crossing between the core and the cladding indices is made possible, despite the low refractive index of the core region and the large holes in the inner cladding, by a small inter-hole distance in the outer cladding region, compared to the inter-hole distance in the inner cladding region (the inter-hole distance in the inner cladding region is three times the inter-hole distance in the outer cladding).","We now turn to a different aspect of the present invention.","Micro-structured cladding structures with a periodically varying refractive index have the potential of exhibiting photonic band gaps.","These are frequency regions where no light transmittance is allowed in the cladding region (see e.g.","Barkou et al., Optical Fiber Communication Conference, FG5-1, pp.","117-119, 1999).","By employing a periodically varying refractive index it is, therefore, possible to obtain guidance of light within low index core regions.","It has been shown both theoretically (Broeng et.","al., Optics Letters, Vol.","25, pp.","96-98, 2000) and experimentally (Cregan et.","al., Science, Vol.","285, pp.","1539-1539, 1999) that this even allows guidance of light within a hollow core, a result that cannot be explained by the theories of standard optical fibers.","Standard fibers, on the other hand, always demand that the core region has the higher refractive index, while the cladding region has the lower refractive index.","The present inventors have realized that one can also have a guided fundamental mode with negative group velocity dispersion with effective mode-index that is below the effective refractive index of the cladding region.","It is important to stress that such a mode cannot be guided according to standard fiber technology.","It is well-known from the theory of photonic band gap fibres that a mode with a mode-index below the effective refractive cladding index can only be guided if the propagation constant of the guided mode lies within a photonic band gap at the particular wavelength.","It is not known (and not at all obvious) that it is possible to make a photonic band gap fibre design where a fundamental mode with large numerical group velocity dispersion exists.","The present inventors have realized a way to obtain this.","As with the designs described above we need a dual concentric core arrangement to obtain such a fundamental mode.","Also the fundamental mode should be guided mainly in the outer core region at long wavelengths and in the inner core region at shorter wavelengths.","As with the fibers described above the fundamental mode exhibits large negative group velocity dispersion only if the slope of the mode-index as a function of the wavelength differ significantly between the two modes when phase matching between the two potential modes occur.","This is an analogue to the case of the preceding fibers.","The core design, however, that may obtain this feature in photonic band gap guiding fibres turn out to be quite different.","Consider the design shown schematically in FIG.","39.The outer cladding consists of air holes 3904 placed on a periodic lattice (a honeycomb lattice in the example shown).","The holes are distributed within a background dielectric material 3903, such as silica.","The enlarged air holes 3902 correspond to the inner cladding of the preceding designs.","They act to lower the effective refractive index of the central core region at long wavelengths so that no guided modes exist in the central core region at long wavelengths.","At shorter wavelengths a mode becomes guided within a photonic band gap.","This may happen even without the central hole 3900 and the high-index ring 3901 being present.","The purpose of 3900 and 3901 will be explained later.","In the design shown in FIG.","40 an outer, low-index core region is included.","It is created by adding air holes 4005 within the six innermost honeycombs.","Notice that this would not act as a core in a standard fiber, since we have effectively lowered the effective index of the outer core region.","In FIG.","40 the outer cladding air holes 4004, the background material 4003, the inner cladding air holes 4002 the high-index ring 4001 and the central air hole 4000 correspond to the ones shown in FIG.","39.In FIG.","41 is shown some effective indices that may be obtained in a fiber with the principal design shown in FIG.","40.The effective refractive index 4100 of the outer cladding lies at the top, corresponding to the fact that no index guided modes exist in the micro-structured fiber, at the wavelengths where the fundamental mode exhibits numerically large negative group velocity dispersion (index-guided modes exist at shorter wavelengths, however, these of not shown for reasons of clarity).","Below 4100 are the photonic band gap edges 4101 and 4102.Any guided mode that is not an index guided mode must have a mode-index between the two photonic band gap edges, since these define the intervals that are not allowed to propagate in the cladding.","A micro-structured fiber-structure such as the one shown in FIG.","39 will have a mode-index as shown by 4105.The purpose of the central air-hole 4000 and the ring with raised refractive index 4001 compared to the background refractive index of the outer cladding is to obtain a mode-index 4105 which traverses the photonic band gap bounded by 4101 and 4102 as steeply as possible.","Notice that the mode-index enters the top of the photonic band gap 4101 at short wavelengths and then descends through the band gap as the wavelength is increased.","A fiber with only the outer core region created by the added air-holes 4005 on the other hand will have a mode-index behaving as 4106.Notice that this mode-index enters the bottom of the photonic band gap 4102 at short wavelengths and then rises through the band gap as the wavelength is increased.","The two types of cores therefore give rise to modes that traverse the photonic band gap quite differently.","The slope of the mode-index as a function of the wavelength of the two modes is therefore quite different when they have an equal mode-index (when they are phase-matched).","Combining the two cores, as shown in FIG.","26 an operation with avoided-crossing is created similar to that referred to with the index-guiding designs.","The fundamental mode has the effective index 4103, while the second mode (that need not carry any energy) has the effective index 4104.The curvature of 4103 corresponds to numerically large negative group velocity dispersion, while the curvature of 4104 corresponds to numerically large positive group velocity dispersion.","Notice that the slope of the two modes 4103 and 4104 tend toward the slope of the two core modes 4105 and 4106 at wavelengths differing from the wavelengths with numerically large group velocity dispersion.","In the wavelength region with numerically large group velocity dispersion the energy of the mode 4103 is moved from the inner core region 4001 to the outer core region 4005 as the wavelength is increased.","Correspondingly, the energy of the mode 4104 is moved from the outer core region 4005 to the inner core region 4001 as the wavelength is increased.","The inventors have realized that a number of advantages exist for designs such as the one shown in FIG.","40, compared to index guiding fiber designs for group velocity dispersion.","It is an advantage that the negative slope of 4102 is larger than the negative slope of 4106.This acts to suppress the mode 4104 if the mode-index of the fundamental mode 4103 is near the photonic band gap bottom, when the fundamental mode exhibits large negative group velocity dispersion.","It is a further advantage that fibers with a design similar to the one shown in FIG.","40 may be designed so that the positive group velocity dispersion of the mode 4104 occurs at wavelengths where the mode-index of 4104 is near the upper photonic band gap edge 4101.This will act to suppress the fundamental mode 4103 so that the positive group velocity dispersion of 4104 can be exploited.","It is therefore an advantage that micro-structured fibers with the design shown in FIG.","40 may be designed to exhibit large positive group velocity dispersion.","This is expected to be of increasing importance as fiber systems with negative group velocity dispersion become more common, in order to e.g.","reduce self-phase modulation in fiber systems.","The present inventors have realized that it is advantageous to use fibers that guide light by the photonic band gap effect for dispersion compensating purposes.","This may seem as a surprise to those skilled in the field, since fibers guiding light by the photonic band gap effect typically have positive dispersion.","However, using a dual concentric core arrangement it becomes possible to have a fundamental super mode with negative group velocity dispersion.","The present inventors have realized that dual concentric core fibers that guide light by the photonic band gap effect are able to have even larger numerical values of the group velocity dispersion than the group velocity dispersion made possible by the prior art.","As indicated by FIG.","41 the large values of group velocity dispersion appear near the wavelength where 4105 crosses 4106.Curve 4105 may be seen as the mode-index of the guided mode within the central core 4001 when the outer core 4005 is omitted, and 4106 may correspondingly be seen as the mode-index of the guided mode within the outer core 4005 when the central core 4001 is omitted.","Using dual concentric cores for group velocity dispersion the amount of group velocity dispersion that can be obtained is directly linked to the difference between the group velocity of the guided modes with a mode-index corresponding to 4105 and 4106.This difference is maximized when the group velocity corresponding to 4105 is minimized while the group velocity corresponding to 4106 is maximized for a given mode-index, β/k.","To investigate the group velocities that can be obtained for guided modes in silica-air fibers, a formula known and understood by those skilled in the art can be applied.","v g = c β k ∫ ∏ E × H * · z ⅆ A ∫ ∏ n 2 E × H * · z ⅆ A Here vg is the group velocity, c is the vacuum velocity of light, β k is the mode-index, E is the electric field, H* is the conjugated magnetic field, n is the refractive index of the material at the given place, II is the cross-section where the mode exist, and z is the length direction of the fiber.","Assuming as an example that the fiber consist of material with n=1 (air) and n=1.45 (silica) we can rewrite this formula as v g = c β k ∫ air E × H * · z ⅆ A + ∫ silica E × H * · z ⅆ A ∫ air E × H * · z ⅆ A + n silica 2 ∫ silica E × H * · z ⅆ A The formula shows that the group velocity of a guided mode depends on the mode-index, β k , as well as the amount of the modal power that propagates in silica, relative to the modal power that propagates in air.","To be more specific the relative amount, fsilica, of the modal power, P = ∫ ∏ E × H * · z ⅆ A , that exist in silica, ∫ silica E × H * · z ⅆ A P , can be used to calculate the group velocity for a guided mode at a given mode-index, such that: v g = c β k 1 + f silica ( n silica 2 - 1 ) From this formula it is seen that a guided mode with the lowest group velocity possible in silica/air will have all its power propagating in silica.","Such a mode will have the group velocity v g = c β k n silica 2 .","The maximum group velocity possible corresponds to a mode with as much of the field as possible propagating in air.","When the mode-index is less than 1 this corresponds to a mode with all its power propagating in air.","Such a mode has the group velocity: v g = c β k , where β k ≤ 1.When the mode-index is greater than 1 the maximum group velocity is equal to the phase-velocity of the mode such that: v g = c β / k , where β 2 ≥ 1.The group velocity dispersion, GVD, of a guided mode may be written as: GVD = ⅆ ( 1 v g ) ⅆ λ where λ is the free space wavelength of the mode.","The amount of group velocity that can be obtained for a given mode-index can therefore be qualitatively estimated by showing the inverse group velocity, 1 v g as a function of the mode-index, β k .","Such a plot is shown in FIG.","42, where 4201 shows the inverse group velocity of a mode that propagates solely in silica, 4202 shows a mode that propagates with the maximum group velocity possible.","4203 is 4202 subtracted from 4201 and therefore shows the propagation time-difference (in pico-seconds, ps) between the slow 4201 and the fast 4202 mode per kilometre of fiber.","Curve 4203 clearly indicates that the propagation time-difference increases strongly as the mode-index, β k is lowered.","Since this propagation time-difference is also a measure of the maximum group velocity dispersion, GVD, that can be obtained by using coupled core regions, curve 4203 therefore also indicates that it is advantageous to have a low mode-index, β k , for dispersion compensating purposes.","Since this is possible by using photonic band gap technology, curve 4203 therefore shows that it is an advantage to use fibers that guide light by the photonic band gap effect for group velocity dispersion.","The present invention therefore also covers photonic crystal fibers with a cladding structure that exhibits photonic band gaps.","It is also important to notice that the present invention may be utilized for fiber applications, wherein fibers according to the present invention guides light in higher order modes.","The dispersion of higher order mode may be even stronger than the dispersion of the fundamental mode.","Apart from the negative dispersion, as a special case, higher order modes in fibers according to the present invention may exhibit very high positive dispersion.","Such fibers, guiding light in a higher order mode, and having large positive dispersion may be used for dispersion compensating schemes for fiber-optical communication systems having a negative dispersion over the transmission length.","Hence, the present invention also covers dispersion compensating fibers that are guiding light in a higher order mode.","Especially, the present invention covers dispersion compensating fibers that are guiding light in a higher order mode with a large positive dispersion.","Typically, it is necessary to employ mode converters in systems that utilize dispersion compensating fibers that guide light in a higher order mode.","Finally, it should be mentioned, that for both single mode dispersion compensation as well as for higher order mode dispersion compensation, the dispersion compensating fibers are typically packed in modules, and the present invention also covers modules that incorporate fibres being disclosed in the present invention.","To fabricate fibers according to the present invention known techniques of stacking and pulling capillary tubes and rods may be used—see for example U.S. Pat.","No.","5,802,236.FIG.","43 shows a schematic example of the cross-section of a preform that may be used to fabricate a fiber with a design as schematically shown in FIG.","12a.","The preform comprises a central rod 4300 with a high-index material in its center.","The rod may have various profiles of the refractive index distribution—for example a step-index profile or a parabolic profile—and it may preferably be fabricated using MCVD techniques.","Surrounding the central rod is, for this example, six capillary tubes 4301.The capillary tubes are surrounded by a number of rods including rods 4302 comprising a high-index material.","The stack of rods and tubes may be put into a larger over-cladding tube 4304 to yield a final preform.","This preform may be drawn into fiber in one or more steps as described for example in U.S. Pat.","No.","5,802,236.In order to realise different size of voids in the first and the second cladding, it may be advantageous to provide a different pressure to the tubes in the two different regions during one or more steps of the fibre drawing process.","In this manner it may for example be advantageous to supply a higher pressure to tubes that define larger voids and a lower pressure to tube that define smaller voids.","Depending on the material, of the fiber, drawing temperatures and speeds may vary significantly and experiments may be needed to determine optimum drawing conditions including pressure.","Alternatively, the preform may be stacked with tubes of different size, for example different inner diameter.","If the preform is drawn at a relatively low temperature, such as around 1850° C.-1900° C. the preform may be drawn to fiber such that the tubes do not collapse.","The tubes with smaller inner diameter will thereby result in voids of smaller size.","Pressure control of the tubes may be advantageous in providing additional control of the void size.","Various manners of applying pressure may be used.","It is for example possible to draw the fiber in two steps, wherein a first step tubes are kept open in both ends and the dimensions of the preform is scaled approximately linearly to a cane during drawing in a fiber drawing tower at a temperature of around 1900° C. to 2000° C. The preform may have a diameter of around 1-50 mm and the cane a diameter of around 1-5 mm.","In the second step, the cane may be drawn to fiber, where one or more regions are connectorized to a pressure-control chamber such that void size in the final fiber may be controlled.","One way of using pressure control for realisation of micro-structured fibers has for example been described by Russell et al.","in WO 00/49436.FIG.","44 shows two microscope pictures (top and bottom illumination with white light) of a fabricated fiber according to the present invention having a design as schematically shown in FIG.","12a.","The fibre has Λ of around 4 μm and is designed to exhibit negative dispersion around 1550 nm.","The figure also shows the measured dispersion characteristics of the fiber (bottom figure).","The fiber is seen to exhibit a dispersion of around −1000 ps/nm/km around 1550 nm—as desirable for dispersion compensating applications.","The fiber further has a strong negative dispersion slope with a relative dispersion slope (RDS)—as desirable for dispersion compensation in WDM systems.","The measured dispersion characteristic is for a mode confined to the central (or first) core of the fiber.","The mode field diameter at 1550 nm is around 4 mm, giving an effective area of around 15 μm2.Hence, the experimental results confirm the simulated results present in the FIGS.","16-23.The core comprises a high-index features being separated from the inner cladding features.","The core features have a refractive index profile with a maximum refractive index of around 1.47 and a diameter of around 0.7 times the center-to-center spacing of the inner cladding features.","The fiber has six high-index features in the outer (or second) core that each has a refractive index profile with a maximum refractive index of around 1.46.The six second core features have a diameter similar to the first core feature.","The features in the first and second core are not directly visible in the microscope picture with top illumination (right, top figure) that mainly serves to show the dimensions of the inner cladding features and their position.","However, using bottom illumination of the fiber and coupling white light to both the first and second core, the high-index features of the two cores become more visible (left, top figure).","It should briefly be mentioned that for operation of the dispersion compensating fibres at near-infrared wavelengths, the mode index of the two cores region is significantly different for light in the visible range (and in according with this and the description of the fibers according to the present invention) and the cores become largely decoupled at the visible where light will be more tightly confined to the high-index features than for the wavelengths of negative dispersion (wavelengths around the previously discussed coupling wavelength regime).","FIG.","45 shows another example of a real fiber according to the present invention.","In comparison with the fiber in FIG.","44, the inner cladding features have a smaller size and the dispersion characteristics is seen to exhibit a less negative minimum, but a broader dispersion “dip”.","This result is also in agreement with the previously presented simulation results.","The fiber presented in FIG.","45 has spectrally broad negative dispersion characteristics and a relatively large RDS, hence making the fiber attractive for dispersion compensation of WDM systems operating around 1550 nm.","FIG.","46 provides a schematic example of a fiber according to the present invention, where the separation between the first core 4600 and the second core 4602 it illustrated.","The separation is characterised by the distance d1-2.The first cladding region comprises low-index features 4601 and the second cladding 4602 is homogeneous.","The figure further illustrates the refractive index profile along one radial direction in the cross-section of the fiber.","The index profile illustrates how the core 4600 is not in contact with the low-index features 4601, but separated by a region 4604.In preferred embodiments, this region comprises a background material being pure silica.","The index profile further shows the radial distance from a center of the second core region to a center of the first core region, d1-2.Twice this distance is also referred to as the diameter of the second core.","FIG.","47 shows a similar illustration as FIG.","46, but for a fiber where the second core is formed from an ensemble of high-index features (4700).","The use of a separation between the first core and the low-index elements has several advantageous—as for example discussed in connection with FIG.","31, and the use of a relatively large diameter of the second core has several advantages as for example discussed in connection with FIG.","18.While FIGS.","46 and 47 illustrates close to parabolic index profiles of the first and second cores, a broad range of other profiles may be used according to the present invention."]],"string":"[\n [\n \"Dual core photonic crystal fibers(pcf) with special dispersion properties\",\n \"An optical fibre for transmitting light, said optical fibre having an axial direction and a cross section perpendicular to said axial direction, said optical fibre comprising: (1) a first core region comprising a first core material having a refractive index Nco,1; (2) a microstructured first cladding region surrounding the first core region, said first cladding region comprising a first cladding material and a plurality of spaced apart first cladding features or elements that are elongated in the fibre axial direction and disposed in the first cladding material, said first cladding material having a refractive index Ncl,1 and each said first cladding feature or element having a refractive index being lower than Ncl,1, whereby a resultant geometrical index Nge,cl, 1?\",\n \"of the first cladding region is lowered compared to Ncl,1; (3) a second core region surrounding said first cladding region, said second core region comprising a second core material having a refractive index Nco,2, and (4) a second cladding region surrounding the second core region, said second cladding region comprising a second cladding material having a refractive index Ncl,2, wherein the first core material, the first cladding material and the first cladding features, the second core material, and the second cladding material are selected and arranged so that Nco,1>Nge,cl,1, Nco,2>Nge,cl,1, and Nco,2>Ncl,2.\"\n ],\n [\n \"1-103.\",\n \"(canceled) 104.An optical fiber for transmitting light, said optical fiber having an axial direction and a cross section perpendicular to said axial direction, said optical fiber comprising: (1) a first core region comprising a first core material having a refractive index Nco,1, (2) a microstructured first cladding region surrounding the first core region, said first cladding region comprising a first cladding material and a plurality of spaced apart first cladding features or elements that are elongated in the fiber axial direction and disposed in the first cladding material, said first cladding material having a refractive index Ncl,1 and each said first cladding feature or element having a refractive index being lower than Ncl,1, whereby a resultant geometrical index Nge,cl,1 of the first cladding region is lowered compared to Ncl,1, (3) a second core region surrounding said first cladding region, said second core region comprising a second core material having a refractive index Nco,2, and (4) a second cladding region surrounding the second core region, said second cladding region comprising a second cladding material having a refractive index Ncl,2, wherein the first core material, the first cladding material and the first cladding features, the second core material, and the second cladding material are selected and arranged so that Nco,1 >Nge,cl,1, Nco,2 >Nge,cl,1, and Nco,2>Ncl,2.105.The optical fiber according to claim 104 wherein the second cladding region is homogeneous.\",\n \"106.The optical fiber according to claim 104 wherein the second core region is micro-structured having a plurality of spaced apart second core features or elements that are elongated in the fiber axial direction and disposed in the second core material, each said second core feature or element having refractive index, Nco,2 being higher than a refractive index, Nco,back,2, of the second core material, whereby a resultant geometrical index Nge,co,2 of the second core region is higher compared to Nco,back,2.107.The optical fiber according to claim 104 wherein Ncl,2 is equal to or lower than Nco,back,2.108.The optical fiber according to claim 104 wherein Ncl,2 is equal to or lower than Ncl,1.109.The optical fiber according to claim 104 wherein the second cladding region is micro-structured having a plurality of spaced apart second cladding features or elements that are elongated in the fiber axial direction and disposed in the second cladding material, each said second cladding feature or element having a refractive index being lower than Ncl,2, whereby a resultant geometrical index Nge,cl,2 of the second cladding region is lowered compared to Ncl,2.110.The optical fiber according to claim 104 wherein the first core region is a solid core region.\",\n \"111.The optical fiber according to claim 104 wherein Nco,1 is larger than Nco,2.112.The optical fiber according to claim 104 wherein Nco,1 is identical to Nco,2.113.The optical fiber according to claim 104 wherein Nco,1 is lower than Nco,2.114.The optical fiber according to claim 104 wherein said second core region is a homogeneous region.\",\n \"115.The optical fiber according to claim 104 wherein the second core region is micro-structured having a plurality of spaced apart second core features or elements that are elongated in the fiber axial direction and disposed in the second core material, each said second core feature or element having a refractive index being lower than Nco,2 whereby a resultant geometrical index Nge,co,2 of the second core region is lowered compared to Nco,2, and wherein Nge,co,2>Nge,cl,1, and Nge,co,2>Nge,cl,2.116.The optical fiber according to claim 106 wherein the second core region further comprises low index second core features.\",\n \"117.The optical fiber according to claim 104 wherein the first cladding features or elements are positioned equidistantly with respect to a center of the fiber.\",\n \"118.The optical fiber according to claim 117 wherein the first cladding features or elements are of similar size.\",\n \"119.The optical fiber according to claim 106 wherein the second core features or elements are positioned equidistantly with respect to a center of the fiber.\",\n \"120.The optical fiber according to claim 119 wherein the second core features are of similar size.\",\n \"121.The optical fiber according to claim 117 wherein the size of the second core features is larger or equal to the size of the first cladding features.\",\n \"122.The optical fiber according to claim 117 wherein the size of the second core features is smaller or larger than the size of the first cladding features.\",\n \"123.The optical fiber according to claim 104 wherein the optical fiber comprises silica glass and Nco,2 is equal to or larger than 1.444.124.The optical fiber according to claim 104 wherein the number of first cladding features is in the range from 6 to 18.125.The optical fiber according to claim 106 wherein the number of second core features is in the range from 6 to 18.126.The optical fiber according to claim 125 wherein the second core features are characterized by a representative center-to-center distance, Λco,2 between two adjacent second core features.\",\n \"127.The optical fiber according to claim 126 wherein the second core features are characterized by a cross-sectional dimension in the range from 0.2Λco,2 to 0.9Λco,2.128.The optical fiber according to claim 104 wherein Nco,1 is in the range from 1.45 to 1.49, and Ncl,1 is in the range from 1.43 to 1.45, and Nco,2 is in the range from 1.44 to 1.47, and Ncl,2 is in the range from 1.43 to 1.45.129.The optical fiber according to claim 104 wherein the first core region is micro-structured having one or more spaced apart first core features or elements that are elongated in the fiber axial direction and disposed in the first core material, each said first core feature having a refractive index being lower than Nco,1, whereby a resultant geometrical index Nge,co,1 of the first core region is lowered compared to Nco,1, and wherein Nge,co,1>Nge,cl,1.130.The optical fiber according to any one of the claim 104 wherein the first core region is micro-structured having one or more spaced apart first core features or elements that are elongated in the fiber axial direction and disposed in the first core material, each said first core feature or element having a refractive index being higher than Nco,1, whereby a resultant geometrical index Nge,co,1 of the first core region is higher compared to Nco,1.131.The optical fiber according to claim 130 wherein the optical fiber comprises one first core feature or element.\",\n \"132.The optical fiber according to claim 131 wherein the first core feature or element is not making contact with the first cladding features or elements.\",\n \"133.The optical fiber according to claim 131 wherein the first core feature or element has a diameter in the range from 2 μm to 4 μm, such as in the range from 2.5 μm to 3.0 μm.\",\n \"134.The optical fiber according to claim 131 wherein the first core feature or element has a diameter in the range from 0.5 to 0.8 times a center-to-center distance between two adjacent first cladding features or elements.\",\n \"135.The optical fiber according to claim 131 wherein the first core region has a background material being identical to the first cladding background material.\",\n \"136.The optical fiber according to claim 129 wherein Nge,co,1 is larger than Nge,co,2.137.The optical fiber according to claim 129 wherein Nge,co,1 is identical to Nge,co,2.138.The optical fiber according to claim 129 wherein Nge,co,1 is lower than Nge,co,2.139.The optical fiber according to claim 104 wherein the first core material is a background material of the first core region, the first cladding material is a background material of the first cladding region, the second core material is a background material of the second core region, and/or the second cladding material is a background material of the second cladding region.\",\n \"140.The optical fiber according to claim 104 wherein the first cladding region has a substantially annular shape in the cross section.\",\n \"141.The optical fiber according to claim 104 wherein the second core region has a substantially annular shape in the cross section.\",\n \"142.The optical fiber according to claim 104 wherein the second cladding region has a substantially annular shape in the cross section.\",\n \"143.The optical fiber according to claim 104 wherein the fiber is dimensioned for transmitting or guiding light of one or more predetermined wavelengths.\",\n \"144.The optical fiber according to claim 143 wherein one of said predetermined wavelengths is in the range from 1.3 μm to 1.7 μm, such as in the range of 1.5 μm to 1.62 μm.\",\n \"145.The optical fiber according to claim 104 wherein said first core region has a first inscribed core diameter, Dco,1, being larger than 3 times said predetermined wavelength, such as larger than 5 times said predetermined wavelength, such as larger than 7 times said predetermined wavelength, such as larger than 10 times said predetermined wavelength.\",\n \"146.The optical fiber according to claim 104 wherein said first core region has a first inscribed core diameter, Dco,1, in the range from 4 μm to 25 μm, such as in the range from 4.0 μm to 5.0 μm, such as in the range from 5.0 μm to 6.5 μm, such as in the range from 6.5 μm to 10.0 μm, such as in the range from 10.0 μm to 25.0 μm.\",\n \"147.The optical fiber according to claim 104 wherein said first core region has a varying refractive index profile, said varying refractive index profile having a highest refractive index equal to Nco,1 and said varying index profile being characterized by an α-profile, where α is in the range from 0 to 100, such as α equal to 2, 3 or higher.\",\n \"148.The optical fiber according to claim 106 wherein said second core features are positioned substantially circularly symmetric with respect to a center of said first core region.\",\n \"149.The optical fiber according to claim 106 wherein said second core features are arranged in a single layer surrounding said first core region, such that a distance from a second core feature to a center of said first core region is substantially identical for all second core features.\",\n \"150.The optical fiber according to claim 106 wherein said second core features are arranged in two or more layers surrounding said first core region.\",\n \"151.An optical fiber according to claim 106 wherein Λco,2 is in the range from 0.2 μm to 20 μm.\",\n \"152.The optical fiber according to claim 104 wherein said first cladding features are positioned substantially circularly symmetric with respect to a center of said first core region.\",\n \"153.The optical fiber according to claim 104 wherein said first cladding features are arranged in a single layer surrounding said first core region, such that a distance from a first cladding feature to a center of said first core region is substantially identical for all first cladding features.\",\n \"154.The optical fiber according to claim 104 wherein said first cladding features are arranged in two or more layers surrounding said first core region.\",\n \"155.The optical fiber according to claim 104 wherein the number of said first cladding features is equal to or larger than 3, such as equal to or larger than 6, such as equal to or larger than 8, such as equal to or larger than 18.156.The optical fiber according to claim 104 wherein said first cladding features have a diameter Dcl,1 and a center-to-center spacing between nearest first cladding features of Λcl,1, and Dcl,1/Λcl,1 is in the range from 0.2 to 0.8, such as from 0.4 to 0.6.157.The optical fiber according to claim 156 wherein Dcl,1 is in the range from 0.1 μm to 10 μm.\",\n \"158.The optical fiber according to claim 156 wherein Λcl,1 is in the range from 0.2 μm to 20 μm, such as from 3.0 μm to 5.0 μm.\",\n \"159.The optical fiber according to claim 106 wherein said second cladding features have a diameter Dcl,2 and a center-to-center spacing between nearest second cladding features of Λcl,2, where Dcl,2/Λcl,2 is in the range from 0.01 to 0.5, such as from 0.1 to 0.2.160.The optical fiber according to claim 159 wherein Dcl,2 is in the range from 0.1 μm to 5 μm.\",\n \"161.The optical fiber according to claim 159 wherein Λcl,2 is in the range from 0.2 μm to 20 μm.\",\n \"162.The optical fiber according to claim 159 wherein Λcl,1 is larger than Λcl,2, such as larger than or equal to 3Λcl,2.163.The optical fiber according to claim 159 wherein Λcl,1 is substantially identical to Λcl,2.164.The optical fiber according to claim 159 wherein Dcl,1 is larger than Dcl,2.165.The optical fiber according to claim 159 wherein Dcl,1/Λcl,1 and Dcl,2/Λcl,2 are substantially identical.\",\n \"166.The optical fiber according to claim 103 wherein said optical fiber comprises one or more glass materials.\",\n \"167.The optical fiber according to claim 166 wherein said optical fiber comprises silica.\",\n \"168.The optical fiber according to claim 103 wherein said optical fiber comprises polymer.\",\n \"169.The optical fiber according to claim 166 wherein said first cladding features, second core features, and/or second cladding features are voids comprising vacuum, air, or another gas.\",\n \"170.The optical fiber according to claim 104 wherein said optical fiber has a non-circular shape of the outer cladding, such as a non-polygonal shape, such as an elliptical shape.\",\n \"171.The optical fiber according to claim 170 wherein said outer shape has a predetermined orientation in the cross section, the predetermined orientation being determined from the position of said first cladding features.\",\n \"172.The optical fiber according to claim 104 wherein said first core region has a non-circular shape in the cross section, such as an elliptical shape, providing a high birefringence in the optical fiber.\",\n \"173.The optical fiber according to claim 104 wherein said first cladding region has a non-circular, inner shape in the cross section, such as an elliptical, inner shape, providing a high birefringence in the optical fiber.\",\n \"174.The optical fiber according to claim 104 wherein said optical fiber guides light at a predetermined wavelength in a single mode.\",\n \"175.The optical fiber according to claim 104 wherein said optical fiber guides light at a predetermined wavelength in a higher order mode.\",\n \"176.The optical fiber according to claim 104 wherein the second core has a diameter larger than 10 μm, such as in the range from 10 μm to 15 μm, such as in the range from 15 μm to 20 μm.\",\n \"177.The optical fiber according to claim 104 wherein the first core has a diameter larger than 3 μm, such as in the range from 4 μm to 8 μm, such as in the range from 4 μm to 6 μm.\",\n \"178.The optical fiber according to claim 104 wherein the first core and the second core are separated by a distance of at least 3 μm, such as at least 4 μm.\",\n \"179.The optical fiber according to claim 104 wherein the first core has a non-zero distance to a low-index feature of the first cladding, such as a distance larger than 0.5 μm, such as larger than 1.0 μm.\",\n \"180.The optical fiber according to claim 104 wherein light is propagating substantially in the first core.\",\n \"181.The optical fiber according to claim 104 wherein light is coupled substantially to the first core.\",\n \"182.The optical fiber according to claim 104 wherein the second core has a larger area than the first core.\",\n \"183.The optical fiber according to claim 104 wherein the second core has an effective refractive index, nco2,eff being larger than (nco,1,eff+ncl,2,eff)/2 or larger than (nco,1,eff+ncl,1,eff)/2, where nco,1,eff is an effective refractive index of the first core region, ncl,1,eff is an effective refractive index of the first cladding region, and ncl,2,eff is an effective refractive index of the second cladding region.\",\n \"184.The optical fiber according to claim 104 wherein the second core has a mean or geometric refractive index, nco2,g being larger than (nco,1,g+ncl,2,g)/2 or larger than (nco,1,g+ncl,1,g)/2, where nco,1,g is a mean or geometric refractive index of the first core region, ncl,1,g is a mean or geometric refractive index of the first cladding region, and ncl,2,g is a mean or geometric refractive index of the second cladding region.\",\n \"185.The optical fiber according to claim 104 wherein said fiber is dimensioned to guide light in a single mode in the first core region at a first wavelength, λ1, being in the range from 1500 to 1640 nm, and said optical fiber exhibits negative dispersion at λ1.186.The optical fiber according to claim 185 wherein the optical fiber is dimensioned to guide light in the second core region for light shorter than a cut-off wavelength, λc, where λc is more than 30 nm shorter than λ1.187.An article comprising a module, said module being employed for dispersion compensation in an optical communication system, said module comprising an optical fiber according to claim 104.188.An optical fiber for transmitting light, said optical fiber having an axial direction and a cross section perpendicular to said axial direction, said optical fiber comprising: (1) a first core region comprising a first core material having a refractive index Nco,1 and at least one first core feature having a refractive being lower than Nco,1, whereby a resultant geometrical index Nge,co,1 of the first core region is lowered compared to Nco,1, (2) a micro-structured first cladding region, said first cladding region surrounding the first core region, and said first cladding region comprising a first cladding material and a plurality of spaced apart first cladding features that are elongated in the fiber axial direction and disposed in the first cladding material, said first cladding material having a refractive index Ncl,1 and each said first cladding feature having a refractive index being lower than Ncl,1, whereby a resultant geometrical index Nge,cl,1 of the first cladding region is lowered compared to Ncl,1, (3) a second core region, said second core region surrounding said first cladding region, and said second core region comprising a second core material having a refractive index Nco,2, and (4) a micro-structured second cladding region, said second cladding region surrounding the second core region, and said second cladding region comprising a second cladding material and a plurality of spaced apart second cladding features that are elongated in the fiber axial direction and disposed in the second cladding material, said second cladding material having a refractive index Ncl,2 and each said second cladding feature having an refractive index being lower than Ncl,2, whereby a resultant geometrical index Nge,cl,2 of the second cladding region is lowered compared to Ncl,2, wherein the first core material, the first cladding material and the first cladding features, the second core material, and the second cladding material and the second cladding features are selected and arranged so that Nco,1>Nge,cl,1, Nco,2>Nge,cl,1, Nco,2>Nge,cl,2, Nge,co,1<Nge,cl,2, and the second cladding features are placed in a substantially two dimensionally periodic structure, whereby the second cladding region exhibits photonic band gap effect.\",\n \"189.The optical fiber according to claim 188 wherein said second cladding features are placed in a substantially two dimensional periodic structure.\",\n \"190.The optical fiber according to claim 188 wherein said second cladding features are placed in a radial periodic structure.\",\n \"191.The optical fiber according to claim 188 wherein the second core has a diameter larger than 10 μm, such as in the range from 10 μm to 15 μm, such as in the range from 15 μm to 20 μm.\",\n \"192.The optical fiber according to claim 188 wherein the first core has a diameter larger than 3 μm, such as in the range from 4 μm to 8 μm, such as in the range from 4 μm to 6 μm.\",\n \"193.The optical fiber according to claim 188 wherein the first core and the second core are separated by a distance of at least 3 μm, such as at least 4 μm.\",\n \"194.The optical fiber according to claim 188 wherein the first core has a non-zero distance to a low-index feature of the first cladding, such as a distance larger than 0.5 μm, such as larger than 1.0 μm.\",\n \"195.The optical fiber according to claim 188 wherein light is propagating substantially in the first core.\",\n \"196.The optical fiber according to claim 188 wherein light is coupled substantially to the first core.\",\n \"197.The optical fiber according to claim 188 wherein the second core has a larger area than the first core.\",\n \"198.The optical fiber according claim 188 wherein the second core has an effective refractive index, nco2,eff being larger than (nco,1,eff+ncl,2,eff)/2 or larger than (nco,1,eff+ncl,1,eff)/2, where nco,1,eff is an effective refractive index of the first core region, ncl,1,eff is an effective refractive index of the first cladding region, and ncl,2,eff is an effective refractive index of the second cladding region.\",\n \"199.The optical fiber according to claim 188 wherein the second core has a mean or geometric refractive index, nco2,g being larger than (nco,1,g+ncl,2,g)/2 or larger than (nco,1,g+ncl,1,g)/2, where nco,1,g is a mean or geometric refractive index of the first core region, ncl,1,g is a mean or geometric refractive index of the first cladding region, and ncl,2,g is a mean or geometric refractive index of the second cladding region.\",\n \"200.The optical fiber according to claim 188 wherein said fiber is dimensioned to guide light in a single mode in the first core region at a first wavelength, λ1, being in the range from 1500 to 1640 nm, and said optical fiber exhibits negative dispersion at λ1.201.The optical fiber according to claim 200 wherein the optical fiber is dimensioned to guide light in the second core region for light shorter than a cut-off wavelength, λc, where λc is more than 30 nm shorter than λ1.202.An article comprising a module, said module being employed for dispersion compensation in an optical communication system, said module comprising an optical fiber according to claim 188.203.An optical transmission line, comprising at least: (1) a length of standard transmission fiber with a positive dispersion and a positive dispersion slope at least at a wavelength, λ1, of 1550 nm, and (2) a length of dispersion compensating fiber, such as a length of dispersion compensating fiber spooled in a fiber module, wherein said dispersion compensating has negative dispersion with a numerical value of the dispersion being larger than 500 ps/nm/km at λ1.204.An optical transmission line, comprising at least: (1) a length of standard transmission fibre with a positive dispersion and a positive dispersion slope at least at a wavelength, λ1, of 1550 nm, and (2) a length of dispersion compensating fibre, such as a length of dispersion compensating fibre according to claim 188 spooled in a fibre module, wherein said dispersion compensating has negative dispersion with a numerical value of the dispersion being larger than 500 ps/nm/km at λ1.205.The optical transmission line according to claim 203 wherein the dispersion compensating fiber has a relative dispersion slope of more than 0.01 nm−1, such as more than 0.02 nm−1.206.An optical fiber for transmitting or guiding light, said optical fiber having an axial direction and a cross section perpendicular to said axial direction, said optical fiber comprising: (1) a first core region comprising a first core material having a refractive index Nco,1, (2) a microstructured first cladding region surrounding the first core region, said first cladding region comprising a first cladding material and a plurality of spaced apart first cladding features or elements that are elongated in the fiber axial direction and disposed in the first cladding material, said first cladding material having a refractive index Ncl,1 and each said first cladding feature or element having a refractive index being lower than Ncl,1, whereby a resultant geometrical index Nge,cl,1 of the first cladding region is lowered compared to Ncl,1, (3) a second core region surrounding said first cladding region, said second core region comprising a second core material having a refractive index Nco,2, and (4) a micro-structured second cladding region surrounding the second core region, said second cladding region comprising a second cladding material and a plurality of spaced apart second cladding features or elements that are elongated in the fiber axial direction and disposed in the second cladding material, said second cladding material having a refractive index Ncl,2 and each said second cladding feature or element having an refractive index being lower than Ncl,2, whereby a resultant geometrical index Nge,cl,2 of the second cladding region is lowered compared to Ncl,2, wherein the first core material, the first cladding material and the first cladding features or elements, the second core material, and the second cladding material and the second cladding features or elements are selected and arranged so that Ncl,1>Nge,cl,1, Nco,2>Nge,cl,1, and Nco,2>Nge,cl,2.207.The optical fiber according to claim 206 wherein the second core region is micro-structured having a plurality of spaced apart second core features or elements that are elongated in the fiber axial direction and disposed in the second core material, each said second core feature or element having a refractive index being higher than Nco,2, whereby a resultant geometrical index Nge,co,2 of the second core region is higher compared to Nco,2.208.The optical fiber according to claim 207 wherein Nco,1 is larger than Nge,co,2.209.The optical fiber according to claim 207 wherein Nco,1 is identical to Nge,co,2.210.The optical fiber according to claim 207 wherein Nco,1 is lower than Nge,co,2.211.The optical fiber according to claim 207 wherein the number of said second core features is equal to or larger than 6, such as equal to or larger than 18.212.The optical fiber according to claim 207 wherein said second core features have a diameter Dco,2 and a center-to-center spacing between nearest second core features of Λco,2, and Dco,2/Λco,2 is in the range from 0.01 to 0.5, such as from 0.1 to 0.2.213.The optical fiber according to claim 212 wherein Dco,2 is in the range from 0.1 μm to 5 μm.\",\n \"214.The optical fiber according to claim 206 wherein said first cladding features have a diameter Dcl,1 and a center-to-center spacing between nearest first cladding features of Λcl,1, and Dcl,1/Λcl,1 is in the range from 0.2 to 0.8, such as from 0.4 to 0.6.215.The optical fiber according to claim 214 wherein Dcl,1/Λcl,1 is larger than Dco,2/Λco,2.216.An optical fiber for transmitting light, said optical fiber having an axial direction and a cross section perpendicular to said axial direction, said optical fiber comprising: (1) a first core region comprising a first core material having a refractive index Nco,1 and at least one first core feature having a refractive being lower than Nco,1, whereby a resultant geometrical index Nge,co,1 of the first core region is lowered compared to Nco,1, (2) a micro-structured first cladding region, said first cladding region surrounding the first core region, and said first cladding region comprising a first cladding material and a plurality of spaced apart first cladding features that are elongated in the fiber axial direction and disposed in the first cladding material, said first cladding material having a refractive index Ncl,1 and each said first cladding feature having a refractive index being lower than Ncl,1, whereby a resultant geometrical index Nge,cl,1 of the first cladding region is lowered compared to Ncl,1, (3) a second core region, said second core region surrounding said first cladding region, and said second core region comprising a second core material having a refractive index Nco,2, and (4) a micro-structured second cladding region, said second cladding region surrounding the second core region, and said second cladding region comprising a second cladding material and a plurality of spaced apart second cladding features that are elongated in the fiber axial direction and disposed in the second cladding material, said second cladding material having a refractive index Ncl,2 and each said second cladding feature having an refractive index being lower than Ncl,2, whereby a resultant geometrical index Nge,cl,2 of the second cladding region is lowered compared to Ncl,2, wherein the first core material, the first cladding material and the first cladding features, the second core material, and the second cladding material and the second cladding features are selected and arranged so that Nco,1>Nge,cl,1, Nco,2>Nge,cl,1, Nco,2>Nge,cl,2, Nge,co,1<Nge,cl,2, and the second cladding features are placed so that the second cladding region exhibits photonic band gap effect.\",\n \"217.The optical fiber according to claim 216 wherein the second core has a diameter larger than 10 μm, such as in the range from 10 μm to 15 μm, such as in the range from 15 μm to 20 μm.\",\n \"218.The optical fiber according to claim 216 wherein the first core has a diameter larger than 3 μm, such as in the range from 4 μm to 8 μm, such as in the range from 4 μm to 6 μm.\",\n \"219.The optical fiber according to claim 216 wherein the first core and the second core are separated by a distance of at least 3 μm, such as at least 4 μm.\",\n \"220.The optical fiber according to claim 216 wherein the first core has a non-zero distance to a low-index feature of the first cladding, such as a distance larger than 0.5 μm, such as larger than 1.0 μm.\",\n \"221.The optical fiber according to claim 216 wherein light is propagating substantially in the first core.\",\n \"222.The optical fiber according to claim 216 wherein light is coupled substantially to the first core.\",\n \"223.The optical fiber according to claim 216 wherein the second core has a larger area than the first core.\",\n \"224.The optical fiber according to claim 216 wherein the second core has an effective refractive index, nco2,eff being larger than (nco,1,eff+ncl,2,eff)/2 or larger than (nco,1,eff+ncl,1,eff)/2, where nco,1,eff is an effective refractive index of the first core region, ncl,1,eff is an effective refractive index of the first cladding region, and ncl,2,eff is an effective refractive index of the second cladding region.\",\n \"225.The optical fiber according to claim 216 wherein the second core has a mean or geometric refractive index, nco2,g being larger than (nco,1,g+ncl,2,g)/2 or larger than (nco,1,g+ncl,1,g)/2, where nco,1,g is a mean or geometric refractive index of the first core region, ncl,1,g is a mean or geometric refractive index of the first cladding region, and ncl,2,g is a mean or geometric refractive index of the second cladding region.\",\n \"226.The optical fiber according to claim 216 wherein said fiber is dimensioned to guide light in a single mode in the first core region at a first wavelength, λ1, being in the range from 1500 to 1640 nm, and said optical fiber exhibits negative dispersion at λ1.227.The optical fiber according to claim 226 wherein the optical fiber is dimensioned to guide light in the second core region for light shorter than a cut-off wavelength, λc, where λc is more than 30 nm shorter than λ1.228.An article comprising a module, said module being employed for dispersion compensation in an optical communication system, said module comprising an optical fiber according to claim 216.229.The article according to claim 228 wherein the dispersion compensating fiber has a relative dispersion slope of more than 0.01 nm−1, such as more than 0.02 nm−1.230.An optical transmission line, comprising at least: (1) a length of standard transmission fiber with a positive dispersion and a positive dispersion slope at least at a wavelength, λ1, of 1550 nm, and (2) a length of dispersion compensating fiber, such as a length of dispersion compensating fiber spooled in a fiber module, wherein said dispersion compensating has negative dispersion with a numerical value of the dispersion being larger than 500 ps/nm/km at λ1.231.An optical transmission line, comprising at least: (1) a length of standard transmission fibre with a positive dispersion and a positive dispersion slope at least at a wavelength, λ1, of 1550 nm, and (2) a length of dispersion compensating fibre, such as a length of dispersion compensating fibre according to claim 216 spooled in a fibre module, wherein said dispersion compensating has negative dispersion with a numerical value of the dispersion being larger than 500 ps/nm/km at λ1.232.The optical transmission line according to claim 230 wherein the dispersion compensating fiber has a relative dispersion slope of more than 0.01 nm−1, such as more than 0.02 nm−1.233.An article comprising a module, said module being employed for dispersion compensation in an optical communication system, said module comprising an optical fiber according to claim 216.234.An optical fiber having an axial direction and a cross section perpendicular to said axial direction, the optical fiber having negative dispersion in the vicinity of a predetermined wavelength and being employed for dispersion compensation; CHARACTERIZED IN THAT; said optical fiber comprises: (1) a first core region being positioned substantially in a center of the optical fiber, the first core region comprising a first core material of refractive index, Nco,1; 2) a first cladding region surrounding the first core region, the first cladding region comprises a multiplicity of spaced apart first cladding features that are elongated in the axial direction and disposed in the first cladding material, said first cladding material having a refractive index Ncl,1; (3) a second core region surrounding the first cladding region, the second core region comprising a second core material of refractive index, Nco,2, and the second core region having a substantially annular shape in the cross section; (4) a second cladding region surrounding the second core region, the second cladding region comprising a second cladding material of refractive index, Ncl,2, and the second cladding region having a substantially annular shape in the cross section.\",\n \"235.The optical fiber according to claim 234 wherein said predetermined wavelength is in a range from 1.3 μm to 1.7 μm, such as from around 1.5 μm to 1.62 μm.\",\n \"236.The optical fiber according to claim 235 wherein Nco,1 is larger than Nco,2.237.The optical fiber according to claim 235 wherein Nco,1 is identical to Nco,2.238.The optical fiber according to claim 235 wherein Nco,1 is lower than Nco,2.239.The optical fiber according to claim 234 wherein said first cladding region has a substantially annular shape in the cross section.\",\n \"240.The optical fiber according to claim 234 wherein said optical fiber has a first inscribed core diameter, Dco,1, being larger than 3 times said predetermined wavelength, such as larger than 5 times said predetermined wavelength, such as larger than 7 times said predetermined wavelength, such as larger than 10 times said predetermined wavelength.\",\n \"241.The optical fiber according to claim 234 wherein Dco,1 is in the range from 4 μm to 25 μm, such as in the range from 4.0 μm to 5.0 μm, such as in the range from 5.0 μm to 6.5 μm, such as in the range from 6.5 μm to 10.0 μm, such as in the range from 10.0 μm to 25.0 μm.\",\n \"242.The optical fiber according to claim 241 wherein said first core region has a varying refractive index profile, said varying refractive index profile having a highest refractive index equal to Nco,1, and said varying index profile being characterized by an α-profile, where α is in the range from 0 to 100, such as α equal to 2, 3 or higher.\",\n \"243.The optical fiber according to claim 234 wherein said second core region is a homogeneous region.\",\n \"244.The optical fiber according to claim 234, wherein said second core region comprises a multiplicity of spaced apart second core features that are elongated in the axial direction and disposed in the second core material.\",\n \"245.The optical fiber according to claim 244 wherein said second core features are positioned substantially circularly symmetric with respect to a center of said first core region.\",\n \"246.The optical fiber according to claim 234 wherein said second core features are arranged in a single layer surrounding said first core region, such that a distance from a second core feature to a center of said first core region is substantially identical for all second core features.\",\n \"247.The optical fiber according to claim 244 wherein said second core features are arranged in two or more layers surrounding said first core region.\",\n \"248.The optical fiber according to claim 244 wherein the number of said second core features is equal to or larger than 6, such as equal to or larger than 18.249.The optical fiber according to claim 244 wherein said second core features have a diameter Dco,2 and a center-to-center spacing between nearest second core features of Λco,2, and Dco,2/Λco,2 is in the range from 0.01 to 0.5, such as from 0.1 to 0.2.250.The optical fiber according to claim 249 wherein Dco,2 is in the range from 0.1 μm to 5 μm.\",\n \"251.The optical fiber according to claim 249 wherein Λco,2 is in the range from 0.2 μm to 20 μm.\",\n \"252.The optical fiber according to claim 234 wherein said first cladding features are positioned substantially circularly symmetric with respect to a center of said first core region.\",\n \"253.The optical fiber according to claim 234 wherein said first cladding features are arranged in a single layer surrounding said first core region, such that a distance from a first cladding feature to a center of said first core region is substantially identical for all first cladding features.\",\n \"254.The optical fiber according to claim 234 wherein said first cladding features are arranged in two or more layers surrounding said first core region.\",\n \"255.The optical fiber according to claim 234 wherein the number of said first cladding features is equal to or larger than 3, such as equal to or larger than 6, such as equal to or larger than 8, such as equal to or larger than 18.256.The optical fiber according to claim 234 wherein said first cladding features have a diameter Dcl,1 and a center-to-center spacing between nearest first cladding features of Λcl,1, and Dcl,1/Λcl,1 is in the range from 0.2 to 0.8, such as from 0.4 to 0.6.257.The optical fiber according to claim 256, wherein Dcl,1 is in the range from 0.1 μm to 10 μm.\",\n \"258.The optical fiber according to claim 256 wherein Λcl,1 is in the range from 0.2 μm to 20 μm.\",\n \"259.The optical fiber according to claim 256 wherein Dcl,1/Λcl,1 is larger than Dco,2/Λco,2.260.The optical fiber according to claim 234 wherein said second cladding region comprises a multiplicity of spaced apart second cladding features that are elongated in the axial direction and disposed in said second cladding material.\",\n \"261.The optical fiber according to claim 260 wherein said second cladding features have a diameter Dcl,2 and a center-to-center spacing between nearest second cladding features of Λcl,2, where Dcl,2/Λcl,2 is in the range from 0.01 to 0.5, such as from 0.1 to 0.2.262.The optical fiber according to claim 261 wherein Dcl,2 is in the range from 0.1 μm to 5 μm.\",\n \"263.The optical fiber according to claim 261 wherein Λcl,2 is in the range from 0.2 μm to 20 μm.\",\n \"264.The optical fiber according to claim 261 wherein Λcl,1 is larger than Λcl,2, such as larger than or equal to 3Λcl,2.265.The optical fiber according to claim 261 wherein Λcl,1 is substantially identical to Λcl,2.266.The optical fiber according to claim 261 wherein Dcl,1 is larger than Dcl,2.267.The optical fiber according to claim 261 wherein Dcl,1/Λcl,1 and Dcl,2/Λcl,2 are substantially identical.\",\n \"268.The optical fiber according to claim 234 wherein said optical fiber comprises one or more glass materials.\",\n \"269.The optical fiber according to claim 268 wherein said optical fiber comprises silica.\",\n \"270.The optical fiber according to claim 234 wherein said optical fiber comprises polymer.\",\n \"271.The optical fiber according to claim 268 wherein said first cladding features, second core features, and/or second cladding features are voids comprising vacuum, air, or another gas.\",\n \"272.The optical fiber according to claim 234 wherein said optical fiber has a non-circular shape of the outer cladding, such as a non-polygonal shape, such as an elliptical shape.\",\n \"273.The optical fiber according to claim 272 wherein said outer shape has a predetermined orientation in the cross section, the predetermined orientation being determined from the position of said first cladding features.\",\n \"274.The optical fiber according to claim 234 wherein said first core region has a non-circular shape in the cross section, such as an elliptical shape, providing a high birefringence in the optical fiber.\",\n \"275.The optical fiber according to claim 234 wherein said first cladding region has a non-circular, inner shape in the cross section, such as an elliptical, inner shape, providing a high birefringence in the optical fiber.\",\n \"276.The optical fiber according to claim 234 wherein said optical fiber guides light at predetermined wavelength in a single mode.\",\n \"277.The optical fiber according to claim 234 wherein said optical fiber guides light at predetermined wavelength in a higher order mode.\",\n \"278.The optical fiber according to claim 234 wherein the second core has a diameter larger than 10 μm, such as in the range from 10 μm to 15 μm, such as in the range from 15 μm to 20 μm.\",\n \"279.The optical fiber according to claim 234 wherein the first core has a diameter larger than 3 μm, such as in the range from 4 μm to 8 μm, such as in the range from 4 μm to 6 μm.\",\n \"280.The optical fiber according to claim 234 wherein the first core and the second core are separated by a distance of at least 3 μm, such as at least 4 μm.\",\n \"281.The optical fiber according to claim 234 wherein the first core has a non-zero distance to a low-index feature of the first cladding, such as a distance larger than 0.5 μm, such as larger than 1.0 μm.\",\n \"282.The optical fiber according to claim 234 wherein light is propagating substantially in the first core.\",\n \"283.The optical fiber according to claim 234 wherein light is coupled substantially to the first core.\",\n \"284.The optical fiber according to claim 234 wherein the second core has a larger area than the first core.\",\n \"285.The optical fiber according to claim 234 wherein the second core has an effective refractive index, nco2,eff being larger than (nco,1,eff+ncl,2,eff)/2 or larger than (nco,1,eff+ncl,1,eff)/2, where nco,1,eff is an effective refractive index of the first core region, ncl,1,eff is an effective refractive index of the first cladding region, and ncl,2,eff is an effective refractive index of the second cladding region.\",\n \"286.The optical fiber according to claim 234 wherein the second core has a mean or geometric refractive index, nco2,g being larger than (nco,1,g+ncl,2,g)/2 or larger than (nco,1,g+ncl,1,g)/2, where nco,1,g is a mean or geometric refractive index of the first core region, ncl,1,g is a mean or geometric refractive index of the first cladding region, and ncl,2,g is a mean or geometric refractive index of the second cladding region.\",\n \"287.The optical fiber according to claim 234 wherein said fiber is dimensioned to guide light in a single mode in the first core region at a first wavelength, λ1, being in the range from 1500 to 1640 nm, and said optical fiber exhibits negative dispersion at λ1.288.The optical fiber according to claim 287 wherein the optical fiber is dimensioned to guide light in the second core region for light shorter than a cut-off wavelength, λc, where λc is more than 30 nm shorter than λ1.289.An article comprising a module, said module being employed for dispersion compensation in an optical communication system, said module comprising an optical fiber according to claim 234.290.An optical transmission line, comprising at least: (1) a length of standard transmission fiber with a positive dispersion and a positive dispersion slope at least at a wavelength, λ1, of 1550 nm, and (2) a length of dispersion compensating fiber, such as a length of dispersion compensating fiber spooled in a fiber module, wherein said dispersion compensating has negative dispersion with a numerical value of the dispersion being larger than 500 ps/nm/km at λ1.291.An optical transmission line, comprising at least: (1) a length of standard transmission fibre with a positive dispersion and a positive dispersion slope at least at a wavelength, λ1, of 1550 nm, and (2) a length of dispersion compensating fibre, such as a length of dispersion compensating fibre according to claim 234 spooled in a fibre module, wherein said dispersion compensating has negative dispersion with a numerical value of the dispersion being larger than 500 ps/nm/km at λ1.292.The optical transmission line according to claim 290 wherein the dispersion compensating fiber has a relative dispersion slope of more than 0.01 nm−1, such as more than 0.02 nm−1 claim 293.An article comprising a module, said module being employed for dispersion compensation in an optical communication system, said module comprising an optical fiber according to claim 234.\"\n ],\n [\n \"<SOH> 1.BACKGROUND OF THE INVENTION <EOH>The present invention relates to new designs of microstructured fibers providing improvements with respect to dispersion compensation (including dispersion slope compensation) for fiber optical communication systems and with respect to non-linear optical fibers for applications at near-infrared wavelengths.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>In a first aspect, the present invention relates to an optical fiber having an axial direction and a cross section perpendicular to said axial direction.\",\n \"The optical fiber has negative dispersion in the vicinity of a predetermined wavelength.\",\n \"The optical fiber is characterised by a number of regions: 1) a first core region being positioned substantially in a center of the optical fiber, the first core region comprising a first core material of refractive index, N co,1 .\",\n \"In the present context, “positioned substantially” means that it is desired to have the first core placed in the center, but manners of production may introduce minor structural deviations such that a center of the first core region and a center of the whole optical fiber may not coincide.\",\n \"Also, the optical fiber may have a non-circular outer shape in the cross section, making the definition of a center of the optical fiber less stringent.\",\n \"In the case of a non-circular outer shape, the center is determined as the center-of-gravity of a two-dimensional element with the same outer shape as the optical fiber.\",\n \"2) a first cladding region surrounding the first core region, the first cladding region comprises a multiplicity of spaced apart first cladding features that are elongated in the axial direction and disposed in the first cladding material, said first cladding material having a refractive index N cl,1 ; 3) a second core region surrounding the first cladding region, the second core region comprising a second core material of refractive index, N co,2 , and the second core region having a substantially annular shape in the cross section.\",\n \"Since the interface between the inner cladding and the outer core may be an interface where at least on one side there is a microstructure (the inner cladding region), the interface may not necessarily be seen as a smooth interface (e.g.\",\n \"in the case of the two regions having identical background refractive indices).\",\n \"Therefore, the shape of the outer core region may not necessarily be a viewed as a smooth ring-like shape—hence the wording “substantially annular shape”; 4) a second cladding region surrounding the second core region, the second cladding region comprising a second cladding material of refractive index, N cl,2 , and the second cladding region having a substantially annular shape in the cross section.\",\n \"In the present context, the wording “substantially annular shape” is used for the same reasons as discussed above.\",\n \"Typically, the predetermined wavelength is in a range from 1.3 μm to 1.7 μm, such as from around 1.5 μm to 1.62 μm.\",\n \"In a preferred embodiment, the optical fiber has a higher refractive index in the first core region than in the second core region.\",\n \"In another preferred embodiment, the optical fiber has identical refractive index in the first core region and in the second core region.\",\n \"This allows for example to manufacture the fiber in a single material, such as e.g.\",\n \"pure silica.\",\n \"In a preferred embodiment, the optical fiber has a lower refractive index in the first core region than in the second core region.\",\n \"This allows for example to have a very large mode field diameter.\",\n \"In another preferred embodiment, the first cladding region has a substantially annular shape in the cross section.\",\n \"The wording “substantially” being used for the same reasons as previously discussed.\",\n \"In another preferred embodiment, the optical fiber has a first inscribed core diameter, D co,1 , being larger than 3 times said predetermined wavelength, such as larger than 5 times said predetermined wavelength, such as larger than 7 times said predetermined wavelength, such as larger than 10 times said predetermined wavelength.\",\n \"In another preferred embodiment, D co,1 is in the range from 4 μm to 25 μm, such as in the range from 4.0 μm to 5.0 μm, such as in the range from 5.0 μm to 6.5 μm, such as in the range from 6.5 μm to 10.0 μm, such as in the range from 10.0 μm to 25.0 μm.\",\n \"In another preferred embodiment, the first core region has a varying refractive index profile, said varying refractive index profile having a highest refractive index equal to N co,1 , and said varying index profile being characterised by an □-profile, where □ is in the range from 0 to 100, such as □ equal to 2, 3 or higher.\",\n \"In another preferred embodiment, the second core region is a homogeneous region.\",\n \"In another preferred embodiment, the second core region is microstructured.\",\n \"This increases the effective refractive index ranges that are available for the second core region without the use of doping.\",\n \"In another preferred embodiment, the microstructured features in the second core are positioned substantially circularly symmetric with respect to a center of said first core region.\",\n \"This is preferred in order to lower polarisation mode dispersion (PMD) in the fiber.\",\n \"An optical fiber according to any one of the preceding claims, wherein said second core features are arranged in a single layer surrounding said first core region, such that a distance from a second core feature to a center of said first core region is substantially identical for all second core features.\",\n \"This is preferred in order to lower PMD, and further preferred for ease of fabrication.\",\n \"In another preferred embodiment, the second core features are arranged in two or more layers surrounding said first core region.\",\n \"This is preferred to reduce bending losses.\",\n \"In another preferred embodiment, the number of second core features is equal to or larger than 6, such as equal to or larger than 18.A high number is preferred to modify the effective refractive index of the second core region.\",\n \"In another preferred embodiment, the second core features have a diameter D co,2 and a center-to-center spacing between nearest second core features of Λ co,2 , and D co,2 /Λ co,2 is in the range from 0.01 to 0.5, such as from 0.1 to 0.2.Generally, the microstructured features in the second core should be relatively small in order to ensure the possibility of the second core to support a limited number of modes.\",\n \"In another preferred embodiment, D co,2 is in the range from 0.1 μm to 5 μm.\",\n \"Typically, this is the design parameter range that provides strongest dispersion.\",\n \"In another preferred embodiment, λ/Λ co,2 is in the range from 0.2 μm to 20 μm.\",\n \"Typically, this is the design parameter range that provides strongest dispersion.\",\n \"In another preferred embodiment, the first cladding features are positioned substantially circularly symmetric with respect to a center of said first core region.\",\n \"This is preferred to lower PMD in the fiber.\",\n \"In another preferred embodiment, the first cladding features are arranged in a single layer surrounding said first core region, such that a distance from a first cladding feature to a center of said first core region is substantially identical for all first cladding features.\",\n \"This is preferred to lower PMD in the fiber.\",\n \"In another preferred embodiment, the said first cladding features are arranged in two or more layers surrounding said first core region.\",\n \"This is preferred to lower bending losses in the fibers and/or to provide stronger dispersion.\",\n \"In another preferred embodiment, the number of said first cladding features is equal to or larger than 3, such as equal to or larger than 6, such as equal to or larger than 8, such as equal to or larger than 18.This range of numbers of features allows for tailoring of the birefringence of the fiber to a large degree.\",\n \"Higher numbers of features may be desired to provide low PMD for example through more circular symmetric feature arrangements.\",\n \"In another preferred embodiment, the first cladding features have a diameter D cl,1 and a center-to-center spacing between nearest first cladding features of Λ cl,1 , and D cl,1 /Λ cl,1 is in the range from 0.2 to 0.8, such as from 0.4 to 0.6.To provide strong dispersion, relatively large features are required.\",\n \"Hence, the features should be large enough to provide a buffer region between the two cores.\",\n \"In another preferred embodiment, D cl,1 is in the range from 0.1 μm to 10 μm.\",\n \"Typically, this range provides the most attractive design parameter range in order to obtain strong dispersion for wavelengths around 1.55 μm.\",\n \"In another preferred embodiment, Λ cl,1 is in the range from 0.2 μm to 20 μm.\",\n \"Typically, this range provides the most attractive design parameter range in order to obtain strong dispersion for wavelengths around 1.55 μm.\",\n \"In another preferred embodiment, D cl,1 /Λ cl,1 is larger than D co,2 /Λ co,2 .\",\n \"This is preferred to ensure that the first cladding region acts as a buffer between the two cores.\",\n \"In another preferred embodiment, the second cladding region comprises a multiplicity of spaced apart second cladding features that are elongated in the axial direction and disposed in said second cladding material.\",\n \"This provides further means for tailoring the dispersion properties of the fiber.\",\n \"In another preferred embodiment, second cladding features have a diameter D cl,2 and a center-to-center spacing between nearest second cladding features of □cl,2, where D cl,2 /Λ cl,2 is in the range from 0.01 to 0.5, such as from 0.1 to 0.2.It is often preferred that these features are comparable to the features of the second core.\",\n \"It may be preferred that the second core is only weakly guiding or has a cut-off close to or around the operational wavelength.\",\n \"Hence the second cladding features may be preferred to have a limited size in order to not provide strong confinement for the second core—or alternatively not to provide confinement of a high number of modes in the second core.\",\n \"In another preferred embodiment, D cl,2 is in the range from 0.1 μm to 5 μm.\",\n \"In accordance with the above-stated preferred embodiment, this is preferred to have the second cladding features being comparable in size to the second core features.\",\n \"Similarly, it is further preferred that Λ cl,2 is in the range from 0.2 μm to 20 μm.\",\n \"In another preferred embodiment, Λcl, 1 is larger than Λ cl,2 .\",\n \"This is preferred in order to allow the first and second cladding regions to have the same background material, but the effective refractive index of the two regions to be different at a given wavelength, such as the predetermined wavelength.\",\n \"In particular, it is preferred to have Λ cl,1 larger or equal to 3Λ cl,2 as such a ratio may easily be realised using well known methods for fabricating microstructured fibers, such as methods employing stacking of capillary tubes.\",\n \"In another preferred embodiment, Λ cl,1 is substantially identical to Λ cl,2 .\",\n \"This may be preferred as similar pitches (i.e.\",\n \"center-to-center features spacing or lattice period in the case of cladding comprising periodic feature arrangements) often allow simple fabrication, thereby improving reproducibility of the fiber.\",\n \"In another preferred embodiment, D cl,1 is larger than D cl,2 .\",\n \"This is preferred in order for the first cladding region to have a lower effective, refractive index than the second cladding.\",\n \"In another preferred embodiment, D cl,1 /Λ cl,1 and D cl,2 /Λ cl,2 are substantially identical.\",\n \"This may be preferred as similar hole and pitch sizes provide the simplest fabrication, thereby improving reproducibility of the fiber.\",\n \"In another preferred embodiment, the optical fiber comprises one or more glass materials.\",\n \"In another preferred embodiment, the optical fiber comprises silica.\",\n \"In another preferred embodiment, the optical fiber comprises polymer.\",\n \"In another preferred embodiment, the first cladding features, second core features, and/or second cladding features are voids comprising vacuum, air, or another gas.\",\n \"In another preferred embodiment, the optical fiber has a non-circular shape of the outer cladding, such as a non-polygonal shape, such as an elliptical shape.\",\n \"This may be advantageous for a range of reasons—for example for fibers with high birefringence (for example for handling splicing issues), or to ensure bending in certain preferred directions.\",\n \"In another preferred embodiment, the outer shape has a predetermined orientation in the cross section, the predetermined orientation being determined from the position of said first cladding features.\",\n \"This may be preferred to further ensure bending in certain preferred directions with respect to the actual directions in the microstructure in and/or around the core region.\",\n \"Also for polarisation maintaining, dispersion compensating fibers this is preferred.\",\n \"In another preferred embodiment, the first core region has a non-circular shape in the cross section, such as an elliptical shape.\",\n \"This is preferred to provide a high birefringence in the optical fiber.\",\n \"In another preferred embodiment, the first cladding region has a non-circular, inner shape in the cross section, such as an elliptical, inner shape, providing a high birefringence in the optical fiber.\",\n \"This is further preferred for polarisation maintaining, dispersion compensating fiber applications.\",\n \"In another preferred embodiment, the optical fiber guides light at predetermined wavelength in a single mode.\",\n \"In another preferred embodiment, the optical fiber guides light at predetermined wavelength in a higher order mode.\",\n \"This is preferred as higher order modes may provide even stronger dispersion than the fundamental mode including the possibility of strong positive dispersion.\",\n \"In another preferred embodiment, the invention covers an article comprising a module, where the module is employed for dispersion compensation in an optical communication system, and the module comprising an optical fiber according to the invention.\",\n \"In a second aspect, the present invention relates to an optical fiber having an axial direction and a cross section perpendicular to said axial direction.\",\n \"The optical fiber has positive dispersion in the vicinity of a predetermined wavelength.\",\n \"The optical fiber is characterised by a number of regions: 1) a first core region being positioned substantially in a center of the optical fiber, the first core region comprising a first core material of refractive index, N co,1 .\",\n \"In the present context, “positioned substantially” means that it is desired to have the first core placed in the center, but manners of production may introduce minor structural deviations such that a center of the first core region and a center of the whole optical fiber may not coincide.\",\n \"Also, the optical fiber may have a non-circular outer shape in the cross section, making the definition of a center of the optical fiber less stringent.\",\n \"In the case of a non-circular outer shape, the center is determined as the center-of-gravity of a two-dimensional element with the same outer shape as the optical fiber.\",\n \"2) a first cladding region surrounding the first core region, the first cladding region comprises a multiplicity of spaced apart first cladding features that are elongated in the axial direction and disposed in the first cladding material, said first cladding material having a refractive index N cl,1 ; 3) a second core region surrounding the first cladding region, the second core region comprising a second core material of refractive index, N co,2 , and the second core region having a substantially annular shape in the cross section.\",\n \"Since the interface between the inner cladding and the outer core may be an interface where at least on one side there is a microstructure (the inner cladding region), the interface may not necessarily be seen as a smooth interface (e.g.\",\n \"in the case of the two regions having identical background refractive indices).\",\n \"Therefore, the shape of the outer core region may not necessarily be viewed as a smooth ring-like shape—hence the wording “substantially annular shape”; 4) a second cladding region surrounding the second core region, the second cladding region comprising a second cladding material of refractive index, N cl,2 , and the second cladding region having a substantially annular shape in the cross section.\",\n \"In the present context, the wording “substantially annular shape” is used for the same reasons as discussed above.\",\n \"In a preferred embodiment, the predetermined wavelength is in a range from 1.3 μm to 1.7 μm, such as from around 1.5 μm to 1.62 μm.\",\n \"In another preferred embodiment, the optical fiber guides light at the predetermined wavelength in a higher-order mode.\",\n \"This is preferred as a higher order mode in a fiber according to the present invention may exhibit a very high positive dispersion or a very high negative dispersion.\",\n \"In a preferred embodiment, the present invention covers an article (this being a fiber optical communication system or parts thereof) that includes a module being employed for dispersion compensation in an optical communication system.\",\n \"The module comprises an optical fiber according to the present invention.\",\n \"In another preferred embodiment, the present invention covers an article that comprises a mode-converter that enables coupling from a conventional single mode fiber to an optical fiber according to the present invention.\",\n \"Finally, it should be mentioned that the present invention covers both fibers that guide light by a modified version of total internal reflection as well as fibers that guide light by photonic band gap effects.\",\n \"Glossary and Definitions For microstructures, a directly measurable quantity is the so-called “filling fraction” that is the volume of disposed features in a microstructure relative to the total volume of a microstructure.\",\n \"For fibers that are invariant in the axial fiber direction, the filling fraction may be determined from direct inspection of the fiber cross-section, e.g.\",\n \"using inspection methods known in the art.\",\n \"In this application we distinguish between “refractive index”, “geometrical index” and “effective index”.\",\n \"The refractive index is the conventional refractive index of a homogeneous material—naturally, this is also used to describe the refractive indices of the various materials themselves in a microstructured medium.\",\n \"The geometrical index of a structure is the geometrically weighted refractive index of the structure.\",\n \"As an example, a micro-structure consisting of air features that occupy 40% of the structure (the air features themselves having a refractive index=1.0) and a background material of silica (the silica background material having a refractive index ≈1.45) has a geometrical index of 0.4×1.0+0.6×1.45=1.27.The procedure of determining the effective refractive index, which for short is referred to as the effective index, of a given micro-structure at a given wavelength is well-known to those skilled in the art (see e.g., Joannopoulos et al., “Photonic Crystals”, Princeton University Press, 1995, or Broeng et al., Optical Fiber Technology, Vol.\",\n \"5, pp.\",\n \"305-330, 1999).\",\n \"Usually, a numerical method capable of solving Maxwell's equation on full vectorial form is required for accurate determination of the effective indices of microstructures.\",\n \"The present invention makes use of employing such a method that has been well documented in the literature (see previous Joannopoulos-reference).\",\n \"In the long-wavelength regime, the effective index is roughly identical to the weighted average of the refractive indices of the constituents of the material, that is, the effective index is close to the geometrical index in this wavelength regime.\"\n ],\n [\n \"1.BACKGROUND OF THE INVENTION The present invention relates to new designs of microstructured fibers providing improvements with respect to dispersion compensation (including dispersion slope compensation) for fiber optical communication systems and with respect to non-linear optical fibers for applications at near-infrared wavelengths.\",\n \"THE TECHNICAL FIELD The development of optical amplifiers operating in the 1550 nm wavelength band of optical communication has during the past ten years formed the basis for a tremendous development of wavelength division multiplexed optical communication systems typically operating in the wavelength interval from 1530 nm to 1610 nm—and rapidly expanding.\",\n \"These amplifiers have to a large degree removed the loss limitations of the optical communication links, and have paved the way for much longer transmission spans operating at significantly higher transmission bit rates (having a much higher transmission capacity).\",\n \"At the same time the development of optical fiber amplifiers at first had the result that systems originally developed to operate in the 1310 nm wavelength band could be upgraded to the 1550 nm band with significant advantages —provided that the dispersion limitations of these systems could be compensated in an efficient manner.\",\n \"To provide a picture of the possibilities and needs for dispersion compensation, it may be noted that when the bit rate is increased to 10 Gbit/s, the dispersion limits the transmission to around 50 km, which obviously raises the need for some sort of dispersion compensation.\",\n \"As described by Grüner-Nielsen et al., ECOC'2000, pp.91-94, non-zero dispersion fibers (NZDF) seems to be the choice for future communication systems due to their low-dispersion and low-non-linear penalties.\",\n \"For bit-rates of 10 Gbit/s NZDFs will not need dispersion compensation before some hundred kilometres of transmission.\",\n \"However, in the future, when the bit rate increases to 40 Gbit/s, dispersion compensation will be needed already after approximately 5 kilometres for non-shifted fibers and after around 30 kilometres when NZDFs are used.\",\n \"It is noteworthy that today several different types of optical transmission fibre are installed with various dispersion properties.\",\n \"However, still a significant part of the installed optical fiber cables make use of non-shifted single-mode fibers, i.e.\",\n \"optical fibres with zero dispersion wavelength at 1310 nm and a dispersion of about 17 ps/km/nm at the wavelength of 1550 nm.\",\n \"The most established dispersion-compensation method today is the use of dispersion compensating fibers (DCFs) operating in the single-mode regime.\",\n \"The use of dispersion compensating fibers was proposed by Lin, Kogelnik, and Cohen, Optics Letters, Vol.5, pp.476-478, 1980.The first DCFs were step index fibers, where the zero dispersion wavelength was moved to wavelengths above 1550 nm by increasing the core index and narrowing the core diameter (see Onishi et al., Electronics Letters, Vol.\",\n \"30, pp.\",\n \"161-163, 1994, and Bjarklev et al., Optics Letters, Vol.19, pp.\",\n \"62-64, 1994).\",\n \"These relatively simple fiber designs allow for control of the dispersion value at a given wavelength, but they do generally not provide the possibility of simultaneous control of the dispersion slope value.\",\n \"It could be argued that one of the ultimate limits for the dispersion compensating properties of this simple step index fiber design may be obtained for a very thin rod of a high index material placed in vacuum (the low-index cladding).\",\n \"At this point it is relevant to refer to the recent development within the area of fiber optics, where a large interest has been pointed towards so-called microstructured fibers, also known as photonic crystal fibers, photonic band gap fibers, hole-assisted fibres and holey fibers.\",\n \"These are fibers providing a number of new properties that are of interest to a wide range of areas such as optical communications, sensor technology, spectroscopy, and medicine (see e.g.\",\n \"Broeng et al., Optical Fiber Technology, Vol.\",\n \"5, pp.\",\n \"305-330, 1999; Broeng et al., Optics Communications, Vol.\",\n \"156, pp.\",\n \"240-244, 1998; Broeng et al, Optics Letters, Vol.\",\n \"25, pp.\",\n \"96-98, 2000; WO 99/64903; WO 99/64904; WO 00/60390; Birks et al., Electronics Letters, Vol.31, pp.1941-1943, October 1995; Knight et al., Journal of the Optical Society of America, A., Vol.15, pp.748-752, March 1998; Knight et al., Optical Materials Vol.\",\n \"11, pp.\",\n \"143-151, 1999; U.S. Pat.\",\n \"No.\",\n \"5,802,236; Monro et al., Journal of Lightwave Technology, Vol.\",\n \"17, pp.\",\n \"1093-1102, 1999; Ferrando et al., Optics Letters, Vol.\",\n \"24, pp.\",\n \"276-278, 1999; WO 00/06506).\",\n \"The fibres are characterized by having a core surrounded by thin, parallel, voids/holes in a background material.\",\n \"The background material is most often a single material such as e.g.\",\n \"silica glass, and the voids/holes commonly contain air or vacuum, but they may also be filled with other glass materials, polymers, liquids, or gasses.\",\n \"Depending on the application, the voids/holes may be periodically or randomly distributed, or they may be distributed in specially designed arrangements incorporating both periodic and non-periodic regions (see e.g.\",\n \"WO 99/64903; WO 99/64904; WO 00/60390; U.S. Pat.\",\n \"No.\",\n \"5,802,236, Monro et al., Optics Letters, Vol.\",\n \"25, pp.\",\n \"206-208, 2000).\",\n \"Now returning to the picture of a single rod of high-index material placed in vacuum, it should be noted that Birks et al.\",\n \"(in IEEE Photonics Technology Letters, Vol.11, pp.\",\n \"674-676, 1999) argued that properties of photonic crystal fibers (PCFs) could be modelled by a silica rod in air.\",\n \"Such approximate calculations indicated that the dispersion of PCFs could exceed −2000 ps/km/nm—or they could compensate (to within + or −0.2%) of the dispersion of 35 times their length of standard fiber over a 100 nm wavelength range.\",\n \"As already mentioned, it is a disadvantage of the step-index fibers that they cannot perform simultaneous compensation of dispersion and dispersion slope, and it is a further disadvantage that very high index contrasts are needed (either obtained through high co-dopant levels or by placing single rods in air).\",\n \"The high-index contrast step-index fibers also have to be designed with very narrow core dimensions in order for the fibers to be single-moded.\",\n \"Regarding non-linearity, this is a further disadvantage especially for systems operating at 40 Gb/s or higher.\",\n \"After now having discussed the simplest approach of obtaining dispersion compensation in standard optical fibers and in photonic crystal fibers, it is useful to look at the further development of standard technology DCFs.\",\n \"As we already have mentioned, one of the key limitations to the simple step-index design was the lack of simultaneous control of dispersion and dispersion slope.\",\n \"One may argue that this limitation simply is due to a quite limited number of design parameters (for a step-index fiber only the core-cladding refractive index difference and the core radius may be selected).\",\n \"The approach in obtaining more design flexibility dates back to the early days of standard fiber technology, and dispersion modification techniques were already studied in the early 1980'ies (see e.g., Monerie, IEEE Journal of Quantum Electronics, Vol.18, pp.535-542, 1982), where double-clad designs (also known as W-fibers) were evaluated.\",\n \"Fibers of this double-clad type also have equivalents realised by photonic crystal fiber technology, and these microstructured fibers have also been explored for dispersion compensating applications.\",\n \"The first microstructured fibers for such applications were disclosed by DiGiovanni et al.\",\n \"(see EP 0 810 453 and U.S. Pat.\",\n \"No.\",\n \"5,802,236).\",\n \"DiGiovanni et al.\",\n \"discussed how microstructured fibers with a cladding divided into two regions could provide improved dispersion characteristics compared to conventional fibers.\",\n \"DiGiovanni et al.\",\n \"used microstructured fibers with an inner, microstructured cladding region having a lower effective refractive index compared to an outer, microstructured cladding region to obtain a dispersion of more than −1500 ps/nm/km.\",\n \"The fibers disclosed by DiGiovanni et al.\",\n \"comprise a single core region (preferably in the centre of the fiber), and it is stated in U.S. Pat.\",\n \"No.\",\n \"5,802,236 that “In general, the microstructure cladding features should be disposed such that the cladding region does not contain any matrix regions of extend in the x-y plane sufficient to act as a secondary core, i.e.\",\n \"to support a propagating radiation mode”.\",\n \"The present inventors have, however, realised that it is advantageous to design microstructured fibres that have at least one region that can support one or more propagating modes other than the mode(s) supported in the central core region.\",\n \"Hence, such a region (that has larger dimensions than the central core and an effective refractive index higher than (No+Nc)/2, where No is the core refractive index and Nc is the effective refractive index of the cladding) that may act as a second core is preferred.\",\n \"In fact, the present inventors have realized that coupling between a mode in the central core region and a mode in the second core is highly advantageous for microstructured optical fibers with special dispersion properties.\",\n \"It is further important to notice that the fiber may be operated to guide light only in a single of the cores—preferably the central core.\",\n \"This desired mechanism may also be described as avoided “crossing” of the core modes, as shall be discussed at a later stage in the detailed description of the invention.\",\n \"It should at this point be noted that strong dispersion compensation generally is obtained by spectrally shifting the effective refractive index of the guided mode(s).\",\n \"This requires refractive index profile control on a sub-wavelength scale, and the effect is closely related to the spatial redistribution of the optical mode as a function of wavelength.\",\n \"When this spatial redistribution of the guided mode takes place in a W-fiber design, the guided mode shifts (for increasing wavelengths) from the high index core to the cladding.\",\n \"This is an effect that generally ties the dispersion compensating effect strongly together with a strong leakage loss and macro bending loss sensitivity very near the desired wavelength for dispersion compensation.\",\n \"Consequently, W-designs have to be made for an extremely narrow selection of production parameters, and this fiber design, therefore, becomes less attractive than designs, where the spatial distribution of optical power is shifted within guided regions of the fiber core.\",\n \"This is also the reason why standard fiber technology generally utilises triple-cladding designs for dispersion compensation (see e.g., Vengsarkar et al., OFC'93, San Jose, USA, Feb. 21-26, 1993, Post deadline paper PD13; Antos, and Smith, IEEE Journal of Lightwave Technology; vol.12, pp.\",\n \"1739-1745, 1994; Vengsarkar et al., OFC'94, paper ThK2, pp.\",\n \"222-227, 1994; Onishi et al., ECOC'94, pp.681-684,1994; Akasaka et al., OFC'96, paper ThA3, pp.\",\n \"201-202,1996).\",\n \"These designs have several advantages over the step index design—among others is negative dispersion slope to provide partly or full dispersion slope compensation.\",\n \"Using standard fiber technology, triple-cladding designs have a narrow high-delta core (i.e.\",\n \"highly doped to obtain a large refractive index difference compared to pure silica) surrounded by a deeply depressed cladding followed by a raised cladding (we here mention the different elements in an order starting at the centre of the fiber cross-section and stepping outwards in radial direction).\",\n \"The third cladding layer is generally made of pure silica.\",\n \"It should also be noted that in order to suppress higher-order mode guidance (lowering the cut-off wavelength of the DCF) an additional depressed cladding layer is often introduced between the raised cladding layer and the outermost cladding.\",\n \"However, for simplicity (and because its influence on the fundamental mode properties often are limited) this fourth cladding layer is often not discussed in large detail.\",\n \"As we have discussed with the more simple standard fiber designs, also triple-cladding design equivalents using microstructured—or PCF—technology have been considered by Fajardo et al.\",\n \"WO 00/16141.Fajardo et al.\",\n \"describe an optical waveguide in which the density and thus the effective refractive indices of the cladding layer is caused to change in a pre-selected way axially along the fiber.\",\n \"The axial change in density of the cladding layer is due to the fraction of the cladding volume that is air or a glass composition different from that of the base cladding glass.\",\n \"Fajardo et al.\",\n \"argue that the axial variation in clad indices changes the signal mode power distribution, thereby changing key waveguide fiber parameters such as magnitude and sign of dispersion, cut-off wavelength and zero dispersion wavelength along the fiber length.\",\n \"It is a disadvantage of the dispersion compensating fibers using standard fiber fabrication techniques (i.e., fibers fabricated from doped silica) that limited dopant levels (given by internal stress, material deposition efficiencies etc.)\",\n \"does not provide as large (negative) dispersion values as other competing techniques.\",\n \"It is a further disadvantage that standard single-mode DCFs typically have significantly smaller spotsize values than the transmission fibers, whose dispersion they are to compensate.\",\n \"The spotsize mismatch generally leads to significant coupling losses, and/or complicated splicing techniques including intermediate fibers, special techniques of material diffusion control etc.\",\n \"When a DCF is used for discrete compensation at the terminal sites, one of the drawbacks of DCFs is that the total link attenuation is increased.\",\n \"The added attenuation must be compensated with additional gain in amplifiers, which degrades the signal-to-noise ratio and increases the cost of the system.\",\n \"To minimise this problem, the DCF should have as high a negative dispersion, DDCF, as possible and as low attenuation, αDCF, as possible.\",\n \"Therefore, a figure of merit (FOM) for dispersion compensating fibers may be defined as the numerical value of DDCF/αDCF.\",\n \"Typical FOM values of presently employed DCFs is around 200 ps/nm/dB, where the attenuation is around 0.5 dB/km and the dispersion is around −100 ps/nm/km.\",\n \"Hence, for a fiber according to the present invention providing a dispersion of −400 ps/nm/km or more negative, loss issues are less restricting than for conventional DCFs—or DCFs according to the present invention may provide higher FOM for similar loss levels.\",\n \"For a dispersion compensating fiber according to the present invention that provides for example a dispersion of −1000 ps/nm/km or more negative, an improved FOM may be achieved for fiber attenuation of up to 5 dB/km compared to conventional DCFs.\",\n \"It should be noted that dispersion compensating fibers fabricated in large-scale production using standard fiber technology normally are designed to have a dispersion of about −100 ps/km/nm at the wavelength of 1550 nm—and having FOM in the order of 200 ps/nm/dB (see e.g., Grüner-Nielsen et al.\",\n \"ECOC'2000, Munich, pp.\",\n \"91-94).\",\n \"However, more negative dispersion values may be obtained—still having good FOM values, and as an example the work of Knudsen et al., LEOS'2000 annual meeting, Paper TuZ2, pp.338-339 may be mentioned.\",\n \"This works demonstrated a dispersion of −190 ps/km/nm for a FOM of 307 ps/nm/dB at 1550 nm.\",\n \"Considering the background of dispersion compensation through the use of specially designed fibers it also becomes relevant to examine the dispersion compensating fibers described by Thyagarajan et al., IEEE Photonics Technology Letters, Vol.8, pp.\",\n \"1510-1512, 1996.In this letter, Thyagarajan et al.\",\n \"describe how very large negative dispersion values (as low as −5100 ps/km/nm) are predicted for fibers consisting of two concentric cores.\",\n \"The desired dispersion compensation is obtained using a centrally placed high-index core (from 2-2.5% relative refractive index difference) and a ring shaped core, chosen independently so that each of the cores supports a single azimuthally symmetric propagating mode at a wavelength of 1550 nm.\",\n \"The idea described by Thyagarajan et al.\",\n \"is that the two-core fiber design then will support two azimuthally symmetric modes (similar to two so-called super modes of a directional coupler), and that these modes are nearly phase matched at 1550 nm.\",\n \"Close to this phase matching wavelength, the mode index of the composite core fiber changes rapidly due to a strong coupling between the two individual modes of the inner core and outer core.\",\n \"Due to a strong refractive index asymmetry between the two cores, there is a rapid change in the slope of the wavelength variation of the fundamental mode index (of the composite structure) leading to a large dispersion around 1550 nm.\",\n \"A fiber of this dual concentric core type has been fabricated, and dispersion results as low as −1800 ps/km/nm at the wavelength of 1550 nm has recently been presented by Auguste et al., IEE Electronics Letters, Vol.\",\n \"36, pp.\",\n \"1689-1691, 2000.Also in this work the functionality of the fiber is described through the coupling between two super modes—one of the super modes having a large negative dispersion and the other having a large positive dispersion—and light being guided through the fiber only in the super mode having negative dispersion.\",\n \"Note also that fibers of this type has been further described by Auguste et al., Optics Communications, Vol.\",\n \"178, pp.\",\n \"71-77, 2000, where the influence of a central dip—generally present in fibers fabricated by the modified chemical vapour deposition (MCVD) method—is studied and demonstrated to be of importance.\",\n \"Considering these dual concentric core fiber designs, it is important to note that the fiber propagation properties also may be interpreted by a more ordinary mode description using linearly polarised (LP) modes in optical fibers.\",\n \"If we look at the dual concentric core designs from this point of view, we find that the LP01 mode (representing one of the previously mentioned super modes) shows a strong negative dispersion.\",\n \"At the same time, the LP02 mode (representing the other of the previously mentioned super modes) shows a strong positive dispersion, and more important—the fiber supports guidance of the LP02-mode.\",\n \"Looking closer at the fiber designs presented by Thyagarajan et al., one also finds that the LP11 mode is strongly guided by the fiber structure.\",\n \"It may be a disadvantage of the dual concentric core designs fabricated by standard fiber technology that the fibers strongly guide several modes.\",\n \"It is a further disadvantage of the dual concentric core designs fabricated by standard fiber technology that very high co-dopant concentrations generally are needed, since it may result in highly increased fiber attenuation, and moreover, it is difficult to maintain a homogeneous, and reproducible dopant level.\",\n \"The latter issue is indicated by Auguste et al.\",\n \"(Optics Communications, Vol.\",\n \"178, pp.\",\n \"71-77, 2000), where it is described how the well known central index dip in MCVD fabricated fibers may introduce strong variations in the spectral location of the dispersion peak (more than 100 nm shifting was reported).\",\n \"It is yet a further disadvantage of the standard technology dual concentric core fibers that the limited index contrast in doped glass technology makes it necessary to operate with ring shaped cores of rather wide outer radii (fibers presented by Thyagarajan et al.\",\n \"and Auguste et al.\",\n \"have radii of the outermost core near 20 microns).\",\n \"This leads to a strongly spatially extended fundamental mode field for wavelengths above 1550 nm, which eventually may result in significant macro-bending sensitivity.\",\n \"High dopant levels are not necessary in PCFs, because index contrast may be obtained through microstructuring.\",\n \"Therefore, the parameter sensitivity may be reduced, and fibers with different guiding properties between the guided and higher order modes may be obtained (mode discrimination is possible).\",\n \"Through a wider range of design possibilities using PCF technology, we may solve the bending loss problem, and at the same time obtain a larger negative dispersion.\",\n \"It is an object of the present invention to provide improved designs and design parameters for microstructured fibers employed for dispersion compensating applications (including dispersion slope compensation).\",\n \"The improvements relates to strong dispersion, control of dispersion slope, low bending losses, large mode-field diameters, lower sensitivity on parameter fluctuations—allowing better reproducibility compared to prior art fibers, and lower requirements on doping levels of silica.\",\n \"Photonic crystal fibres (PCFs) having multiple cladding layers have been described by Hasegawa et al.\",\n \"in EP 1 118 887 A2.Hasegawa et al.\",\n \"claim an optical fiber including a core region and cladding regions of not less than three layers, which surround the core region in order, wherein at least one of the cladding regions has lower mean refractive index than both adjacent cladding regions, and at least one cladding region is provided with a plurality of sub medium regions each having a refractive index lower than a main medium constituting this cladding region.\",\n \"The present inventors have, however, realised that it is advantageous to design microstructured fibers that have at least one region that can support one or more propagating modes other than the mode(s) supported in the central core region.\",\n \"It is a disadvantage of the fibers described by Hasegawa et al.\",\n \"that they do not include such a region that may act as a core in its own right supporting a propagating mode, since it may limit the possible magnitude of the obtainable dispersion.\",\n \"Furthermore, the present inventors have realised that it may be advantageous not to have a cladding region with a lower effective refractive index than two adjacent cladding regions.\",\n \"In fact the present inventors have realised that it is an advantage to have a region with a higher effective refractive index than two adjacent cladding—such a region is here termed “second core”.\",\n \"The fibres disclosed by Hasegawa et al.\",\n \"exhibit a negative dispersion of around −200 ps/km/nm, whereas the micro-structured fibers disclosed by the present inventors provides negative dispersion in excess of −10000 ps/nm/km.\",\n \"In the patent application by Hasegawa et al., it is given that the mean refractive index n3 of the outer cladding region obeys the relation n2>n3, where n2 is the refractive index of the second inner cladding.\",\n \"Hasegawa et al.\",\n \"obtains this relation by incorporating low-index features in cladding region 3.This is in contrast to a number of preferred embodiments of the present invention, where the outer cladding consists of a homogeneous material.\",\n \"SUMMARY OF THE INVENTION In a first aspect, the present invention relates to an optical fiber having an axial direction and a cross section perpendicular to said axial direction.\",\n \"The optical fiber has negative dispersion in the vicinity of a predetermined wavelength.\",\n \"The optical fiber is characterised by a number of regions: 1) a first core region being positioned substantially in a center of the optical fiber, the first core region comprising a first core material of refractive index, Nco,1.In the present context, “positioned substantially” means that it is desired to have the first core placed in the center, but manners of production may introduce minor structural deviations such that a center of the first core region and a center of the whole optical fiber may not coincide.\",\n \"Also, the optical fiber may have a non-circular outer shape in the cross section, making the definition of a center of the optical fiber less stringent.\",\n \"In the case of a non-circular outer shape, the center is determined as the center-of-gravity of a two-dimensional element with the same outer shape as the optical fiber.\",\n \"2) a first cladding region surrounding the first core region, the first cladding region comprises a multiplicity of spaced apart first cladding features that are elongated in the axial direction and disposed in the first cladding material, said first cladding material having a refractive index Ncl,1; 3) a second core region surrounding the first cladding region, the second core region comprising a second core material of refractive index, Nco,2, and the second core region having a substantially annular shape in the cross section.\",\n \"Since the interface between the inner cladding and the outer core may be an interface where at least on one side there is a microstructure (the inner cladding region), the interface may not necessarily be seen as a smooth interface (e.g.\",\n \"in the case of the two regions having identical background refractive indices).\",\n \"Therefore, the shape of the outer core region may not necessarily be a viewed as a smooth ring-like shape—hence the wording “substantially annular shape”; 4) a second cladding region surrounding the second core region, the second cladding region comprising a second cladding material of refractive index, Ncl,2, and the second cladding region having a substantially annular shape in the cross section.\",\n \"In the present context, the wording “substantially annular shape” is used for the same reasons as discussed above.\",\n \"Typically, the predetermined wavelength is in a range from 1.3 μm to 1.7 μm, such as from around 1.5 μm to 1.62 μm.\",\n \"In a preferred embodiment, the optical fiber has a higher refractive index in the first core region than in the second core region.\",\n \"In another preferred embodiment, the optical fiber has identical refractive index in the first core region and in the second core region.\",\n \"This allows for example to manufacture the fiber in a single material, such as e.g.\",\n \"pure silica.\",\n \"In a preferred embodiment, the optical fiber has a lower refractive index in the first core region than in the second core region.\",\n \"This allows for example to have a very large mode field diameter.\",\n \"In another preferred embodiment, the first cladding region has a substantially annular shape in the cross section.\",\n \"The wording “substantially” being used for the same reasons as previously discussed.\",\n \"In another preferred embodiment, the optical fiber has a first inscribed core diameter, Dco,1, being larger than 3 times said predetermined wavelength, such as larger than 5 times said predetermined wavelength, such as larger than 7 times said predetermined wavelength, such as larger than 10 times said predetermined wavelength.\",\n \"In another preferred embodiment, Dco,1 is in the range from 4 μm to 25 μm, such as in the range from 4.0 μm to 5.0 μm, such as in the range from 5.0 μm to 6.5 μm, such as in the range from 6.5 μm to 10.0 μm, such as in the range from 10.0 μm to 25.0 μm.\",\n \"In another preferred embodiment, the first core region has a varying refractive index profile, said varying refractive index profile having a highest refractive index equal to Nco,1, and said varying index profile being characterised by an □-profile, where □ is in the range from 0 to 100, such as □ equal to 2, 3 or higher.\",\n \"In another preferred embodiment, the second core region is a homogeneous region.\",\n \"In another preferred embodiment, the second core region is microstructured.\",\n \"This increases the effective refractive index ranges that are available for the second core region without the use of doping.\",\n \"In another preferred embodiment, the microstructured features in the second core are positioned substantially circularly symmetric with respect to a center of said first core region.\",\n \"This is preferred in order to lower polarisation mode dispersion (PMD) in the fiber.\",\n \"An optical fiber according to any one of the preceding claims, wherein said second core features are arranged in a single layer surrounding said first core region, such that a distance from a second core feature to a center of said first core region is substantially identical for all second core features.\",\n \"This is preferred in order to lower PMD, and further preferred for ease of fabrication.\",\n \"In another preferred embodiment, the second core features are arranged in two or more layers surrounding said first core region.\",\n \"This is preferred to reduce bending losses.\",\n \"In another preferred embodiment, the number of second core features is equal to or larger than 6, such as equal to or larger than 18.A high number is preferred to modify the effective refractive index of the second core region.\",\n \"In another preferred embodiment, the second core features have a diameter Dco,2 and a center-to-center spacing between nearest second core features of Λco,2, and Dco,2/Λco,2 is in the range from 0.01 to 0.5, such as from 0.1 to 0.2.Generally, the microstructured features in the second core should be relatively small in order to ensure the possibility of the second core to support a limited number of modes.\",\n \"In another preferred embodiment, Dco,2 is in the range from 0.1 μm to 5 μm.\",\n \"Typically, this is the design parameter range that provides strongest dispersion.\",\n \"In another preferred embodiment, λ/Λco,2 is in the range from 0.2 μm to 20 μm.\",\n \"Typically, this is the design parameter range that provides strongest dispersion.\",\n \"In another preferred embodiment, the first cladding features are positioned substantially circularly symmetric with respect to a center of said first core region.\",\n \"This is preferred to lower PMD in the fiber.\",\n \"In another preferred embodiment, the first cladding features are arranged in a single layer surrounding said first core region, such that a distance from a first cladding feature to a center of said first core region is substantially identical for all first cladding features.\",\n \"This is preferred to lower PMD in the fiber.\",\n \"In another preferred embodiment, the said first cladding features are arranged in two or more layers surrounding said first core region.\",\n \"This is preferred to lower bending losses in the fibers and/or to provide stronger dispersion.\",\n \"In another preferred embodiment, the number of said first cladding features is equal to or larger than 3, such as equal to or larger than 6, such as equal to or larger than 8, such as equal to or larger than 18.This range of numbers of features allows for tailoring of the birefringence of the fiber to a large degree.\",\n \"Higher numbers of features may be desired to provide low PMD for example through more circular symmetric feature arrangements.\",\n \"In another preferred embodiment, the first cladding features have a diameter Dcl,1 and a center-to-center spacing between nearest first cladding features of Λcl,1, and Dcl,1/Λcl,1 is in the range from 0.2 to 0.8, such as from 0.4 to 0.6.To provide strong dispersion, relatively large features are required.\",\n \"Hence, the features should be large enough to provide a buffer region between the two cores.\",\n \"In another preferred embodiment, Dcl,1 is in the range from 0.1 μm to 10 μm.\",\n \"Typically, this range provides the most attractive design parameter range in order to obtain strong dispersion for wavelengths around 1.55 μm.\",\n \"In another preferred embodiment, Λcl,1 is in the range from 0.2 μm to 20 μm.\",\n \"Typically, this range provides the most attractive design parameter range in order to obtain strong dispersion for wavelengths around 1.55 μm.\",\n \"In another preferred embodiment, Dcl,1/Λcl,1 is larger than Dco,2/Λco,2.This is preferred to ensure that the first cladding region acts as a buffer between the two cores.\",\n \"In another preferred embodiment, the second cladding region comprises a multiplicity of spaced apart second cladding features that are elongated in the axial direction and disposed in said second cladding material.\",\n \"This provides further means for tailoring the dispersion properties of the fiber.\",\n \"In another preferred embodiment, second cladding features have a diameter Dcl,2 and a center-to-center spacing between nearest second cladding features of □cl,2, where Dcl,2/Λcl,2 is in the range from 0.01 to 0.5, such as from 0.1 to 0.2.It is often preferred that these features are comparable to the features of the second core.\",\n \"It may be preferred that the second core is only weakly guiding or has a cut-off close to or around the operational wavelength.\",\n \"Hence the second cladding features may be preferred to have a limited size in order to not provide strong confinement for the second core—or alternatively not to provide confinement of a high number of modes in the second core.\",\n \"In another preferred embodiment, Dcl,2 is in the range from 0.1 μm to 5 μm.\",\n \"In accordance with the above-stated preferred embodiment, this is preferred to have the second cladding features being comparable in size to the second core features.\",\n \"Similarly, it is further preferred that Λcl,2 is in the range from 0.2 μm to 20 μm.\",\n \"In another preferred embodiment, Λcl,1 is larger than Λcl,2.This is preferred in order to allow the first and second cladding regions to have the same background material, but the effective refractive index of the two regions to be different at a given wavelength, such as the predetermined wavelength.\",\n \"In particular, it is preferred to have Λcl,1 larger or equal to 3Λcl,2 as such a ratio may easily be realised using well known methods for fabricating microstructured fibers, such as methods employing stacking of capillary tubes.\",\n \"In another preferred embodiment, Λcl,1 is substantially identical to Λcl,2.This may be preferred as similar pitches (i.e.\",\n \"center-to-center features spacing or lattice period in the case of cladding comprising periodic feature arrangements) often allow simple fabrication, thereby improving reproducibility of the fiber.\",\n \"In another preferred embodiment, Dcl,1 is larger than Dcl,2.This is preferred in order for the first cladding region to have a lower effective, refractive index than the second cladding.\",\n \"In another preferred embodiment, Dcl,1/Λcl,1 and Dcl,2/Λcl,2 are substantially identical.\",\n \"This may be preferred as similar hole and pitch sizes provide the simplest fabrication, thereby improving reproducibility of the fiber.\",\n \"In another preferred embodiment, the optical fiber comprises one or more glass materials.\",\n \"In another preferred embodiment, the optical fiber comprises silica.\",\n \"In another preferred embodiment, the optical fiber comprises polymer.\",\n \"In another preferred embodiment, the first cladding features, second core features, and/or second cladding features are voids comprising vacuum, air, or another gas.\",\n \"In another preferred embodiment, the optical fiber has a non-circular shape of the outer cladding, such as a non-polygonal shape, such as an elliptical shape.\",\n \"This may be advantageous for a range of reasons—for example for fibers with high birefringence (for example for handling splicing issues), or to ensure bending in certain preferred directions.\",\n \"In another preferred embodiment, the outer shape has a predetermined orientation in the cross section, the predetermined orientation being determined from the position of said first cladding features.\",\n \"This may be preferred to further ensure bending in certain preferred directions with respect to the actual directions in the microstructure in and/or around the core region.\",\n \"Also for polarisation maintaining, dispersion compensating fibers this is preferred.\",\n \"In another preferred embodiment, the first core region has a non-circular shape in the cross section, such as an elliptical shape.\",\n \"This is preferred to provide a high birefringence in the optical fiber.\",\n \"In another preferred embodiment, the first cladding region has a non-circular, inner shape in the cross section, such as an elliptical, inner shape, providing a high birefringence in the optical fiber.\",\n \"This is further preferred for polarisation maintaining, dispersion compensating fiber applications.\",\n \"In another preferred embodiment, the optical fiber guides light at predetermined wavelength in a single mode.\",\n \"In another preferred embodiment, the optical fiber guides light at predetermined wavelength in a higher order mode.\",\n \"This is preferred as higher order modes may provide even stronger dispersion than the fundamental mode including the possibility of strong positive dispersion.\",\n \"In another preferred embodiment, the invention covers an article comprising a module, where the module is employed for dispersion compensation in an optical communication system, and the module comprising an optical fiber according to the invention.\",\n \"In a second aspect, the present invention relates to an optical fiber having an axial direction and a cross section perpendicular to said axial direction.\",\n \"The optical fiber has positive dispersion in the vicinity of a predetermined wavelength.\",\n \"The optical fiber is characterised by a number of regions: 1) a first core region being positioned substantially in a center of the optical fiber, the first core region comprising a first core material of refractive index, Nco,1.In the present context, “positioned substantially” means that it is desired to have the first core placed in the center, but manners of production may introduce minor structural deviations such that a center of the first core region and a center of the whole optical fiber may not coincide.\",\n \"Also, the optical fiber may have a non-circular outer shape in the cross section, making the definition of a center of the optical fiber less stringent.\",\n \"In the case of a non-circular outer shape, the center is determined as the center-of-gravity of a two-dimensional element with the same outer shape as the optical fiber.\",\n \"2) a first cladding region surrounding the first core region, the first cladding region comprises a multiplicity of spaced apart first cladding features that are elongated in the axial direction and disposed in the first cladding material, said first cladding material having a refractive index Ncl,1; 3) a second core region surrounding the first cladding region, the second core region comprising a second core material of refractive index, Nco,2, and the second core region having a substantially annular shape in the cross section.\",\n \"Since the interface between the inner cladding and the outer core may be an interface where at least on one side there is a microstructure (the inner cladding region), the interface may not necessarily be seen as a smooth interface (e.g.\",\n \"in the case of the two regions having identical background refractive indices).\",\n \"Therefore, the shape of the outer core region may not necessarily be viewed as a smooth ring-like shape—hence the wording “substantially annular shape”; 4) a second cladding region surrounding the second core region, the second cladding region comprising a second cladding material of refractive index, Ncl,2, and the second cladding region having a substantially annular shape in the cross section.\",\n \"In the present context, the wording “substantially annular shape” is used for the same reasons as discussed above.\",\n \"In a preferred embodiment, the predetermined wavelength is in a range from 1.3 μm to 1.7 μm, such as from around 1.5 μm to 1.62 μm.\",\n \"In another preferred embodiment, the optical fiber guides light at the predetermined wavelength in a higher-order mode.\",\n \"This is preferred as a higher order mode in a fiber according to the present invention may exhibit a very high positive dispersion or a very high negative dispersion.\",\n \"In a preferred embodiment, the present invention covers an article (this being a fiber optical communication system or parts thereof) that includes a module being employed for dispersion compensation in an optical communication system.\",\n \"The module comprises an optical fiber according to the present invention.\",\n \"In another preferred embodiment, the present invention covers an article that comprises a mode-converter that enables coupling from a conventional single mode fiber to an optical fiber according to the present invention.\",\n \"Finally, it should be mentioned that the present invention covers both fibers that guide light by a modified version of total internal reflection as well as fibers that guide light by photonic band gap effects.\",\n \"Glossary and Definitions For microstructures, a directly measurable quantity is the so-called “filling fraction” that is the volume of disposed features in a microstructure relative to the total volume of a microstructure.\",\n \"For fibers that are invariant in the axial fiber direction, the filling fraction may be determined from direct inspection of the fiber cross-section, e.g.\",\n \"using inspection methods known in the art.\",\n \"In this application we distinguish between “refractive index”, “geometrical index” and “effective index”.\",\n \"The refractive index is the conventional refractive index of a homogeneous material—naturally, this is also used to describe the refractive indices of the various materials themselves in a microstructured medium.\",\n \"The geometrical index of a structure is the geometrically weighted refractive index of the structure.\",\n \"As an example, a micro-structure consisting of air features that occupy 40% of the structure (the air features themselves having a refractive index=1.0) and a background material of silica (the silica background material having a refractive index ≈1.45) has a geometrical index of 0.4×1.0+0.6×1.45=1.27.The procedure of determining the effective refractive index, which for short is referred to as the effective index, of a given micro-structure at a given wavelength is well-known to those skilled in the art (see e.g., Joannopoulos et al., “Photonic Crystals”, Princeton University Press, 1995, or Broeng et al., Optical Fiber Technology, Vol.\",\n \"5, pp.\",\n \"305-330, 1999).\",\n \"Usually, a numerical method capable of solving Maxwell's equation on full vectorial form is required for accurate determination of the effective indices of microstructures.\",\n \"The present invention makes use of employing such a method that has been well documented in the literature (see previous Joannopoulos-reference).\",\n \"In the long-wavelength regime, the effective index is roughly identical to the weighted average of the refractive indices of the constituents of the material, that is, the effective index is close to the geometrical index in this wavelength regime.\",\n \"3.BRIEF DESCRIPTION OF THE DRAWINGS In the following, by way of examples only, the invention is further disclosed with detailed description of preferred embodiments.\",\n \"Reference is made to the drawings in which: FIG.\",\n \"1a (prior art) shows schematically the refractive index profile of a conventional step-index fiber.\",\n \"FIG.\",\n \"1b (prior art) shows schematically the cross-section of an equivalent index-guiding photonic crystal fiber.\",\n \"FIG.\",\n \"2a (prior art) shows schematically the refractive index profile of a conventional W-type fiber generally suggested for dispersion management.\",\n \"FIG.\",\n \"2b (prior art) shows schematically the cross-section of a microstructured fiber with two different air-void dimensions used to obtain an effective index profile of the W-type.\",\n \"FIG.\",\n \"3a (prior art) shows schematically the refractive index profile of a conventional triple-clad fiber of a type generally used for standard dispersion compensating fibers.\",\n \"FIG.\",\n \"3b (prior art) shows schematically the cross-section of a microstructured fiber with three different air-void dimensions used to obtain an effective index profile of the triple-clad type.\",\n \"FIG.\",\n \"4 (prior art) shows the effective index of micro-structures comprising air holes as a function of normalised wavelength for different air hole dimensions for.\",\n \"FIG.\",\n \"5 (prior art) shows schematically the cross-section of a microstructured fiber for dispersion compensation, the fiber having a doped central core surrounded by an annular inner cladding section containing larger air holes than those used to form the microstructured outer cladding; The fiber comprises further a high-index features in the center of the core.\",\n \"FIG.\",\n \"6 (prior art) shows the effective mode indices of periodic outer cladding regions and of a hypothetical honeycomb structures shown in FIG.\",\n \"5 (representing the central fiber region) as a function of the normalised wavelength.\",\n \"Curves are shown for honeycomb structures with different air-hole dimensions.\",\n \"FIG.\",\n \"7 shows the effective mode indices of periodic outer cladding regions and of a hypothetical honeycomb structures (representing the central fiber region) as a function of the normalised wavelength.\",\n \"Curves are shown for different doping levels in the central core, and for different honeycomb air-hole dimensions.\",\n \"FIG.\",\n \"8 (embodiment) shows schematically the cross-section of microstructured fiber for dispersion compensation, the fiber having a doped central core surrounded by an annular inner cladding section containing larger air holes than those used to form the microstructured outer cladding.\",\n \"Between the inner and outer cladding regions, a higher effective index is obtained in an annular segment, where air holes have been removed (compared to the outer cladding).\",\n \"FIG.\",\n \"9 shows a simulation of the mode indices for a fiber according to the present invention.\",\n \"FIG.\",\n \"10 shows a simulation of the dispersion of a dispersion-compensating fiber according to the present invention.\",\n \"FIG.\",\n \"11 shows the field distribution of the fundamental mode for a microstructured fiber according to the invention.\",\n \"FIG.\",\n \"12 shows schematically a fiber according to the present invention where the outer cladding is homogeneous and a) the outer core comprises high-index features or b) the outer core comprises an annular high-index region.\",\n \"FIG.\",\n \"13 shows qualitatively the operation of a dispersion compensation fiber according to the present invention that may have large negative dispersion and low bending losses.\",\n \"FIG.\",\n \"14 shows schematically a preferred positioning of low-index features in the inner cladding and of high-index features in the outer core with respect to each other.\",\n \"FIG.\",\n \"15 shows schematically a fiber according to the present invention where the outer cladding is homogeneous and comprises a material different from the background material of the inner cladding and of the outer core.\",\n \"Typically, the outer cladding material has a lower refractive index than the background material of the inner cladding and of the outer core.\",\n \"FIG.\",\n \"16 shows a simulation of the mode indices for another fiber according to the present invention.\",\n \"FIG.\",\n \"17 shows a simulation of the dispersion of a range of fibers according to the present invention.\",\n \"FIG.\",\n \"18 shows a simulation of the wavelength of strongest negative dispersion and the corresponding hole spacing for a range of fibers according to the present invention designed for operation around 1550 nm.\",\n \"FIG.\",\n \"19 shows a simulation of the strongest negative dispersion value of a range of fibers according to the present invention.\",\n \"FIG.\",\n \"20 shows a simulation of the width of the dispersion “dip” of a range of fibers according to the present invention.\",\n \"FIG.\",\n \"21 shows a simulation of the dispersion and relative dispersion slope for a fiber according to the present invention.\",\n \"FIG.\",\n \"22 shows a simulation of the dispersion and relative dispersion slope for another fiber according to the present invention.\",\n \"FIG.\",\n \"23 shows a simulation of the dispersion and relative dispersion slope for yet another fiber according to the present invention.\",\n \"FIG.\",\n \"24 illustrates the operation of another fiber according to the present invention.\",\n \"The figure illustrates the possibility of using higher order modes in the central core for dispersion compensating applications for fibers according to the present invention.\",\n \"FIG.\",\n \"25 shows a schematic example of another fiber according to the present invention.\",\n \"FIG.\",\n \"26 illustrates the operation of a fiber according to the present invention having an outer core with the same refractive index as the refractive index of the inner cladding region.\",\n \"FIG.\",\n \"27 illustrates the operation of a fiber according to the present invention being comparable to the fiber in the previous figure, but with a larger size of the outer core.\",\n \"FIG.\",\n \"28 shows a simulation of the dispersion and the effective area of the fiber of the previous figure.\",\n \"FIG.\",\n \"29 shows a simulation of the dispersion and the effective area of a fiber according to the present invention having a high-index core not being in contact with the inner cladding.\",\n \"FIG.\",\n \"30 shows a comparison of the effective area of the fibers in the two previous figures.\",\n \"FIG.\",\n \"31 shows a comparison of the dispersion of the fibers in FIGS.\",\n \"28 and 29.FIG.\",\n \"32 shows schematically the cross-section of a micro-structured fiber for dispersion compensation, the fiber has a core with a doped feature, the core being surrounded by an annular inner cladding section containing more closely placed air holes than those used to form the microstructured outer cladding.\",\n \"Between the inner and outer cladding regions, a higher effective index is obtained in an annular segment, where air holes have been removed (compared to the outer cladding).\",\n \"FIG.\",\n \"33 shows schematically the cross-section of a micro-structured fiber for dispersion compensation, the fiber having a core with a doped portion and the core is surrounded by an annular inner cladding section containing more closely placed air holes than those used to form the microstructured outer cladding.\",\n \"Compared to the previous figure, additional air holes have been removed in the outer core region.\",\n \"FIG.\",\n \"34 schematically shows the cross-section of a micro-structured fiber according to the invention, in which the outer core comprises high-index features.\",\n \"FIG.\",\n \"35 schematically shows the cross-section of a micro-structured fiber according to the invention, in which the outer core comprises a single annular feature with a higher refractive index than the outer cladding background material.\",\n \"FIG.\",\n \"36 schematically shows the cross-section of a micro-structured fiber according to the invention, in which the outer cladding is formed by air holes placed in concentric circles.\",\n \"FIG.\",\n \"37 schematically shows the cross-section of a micro-structured fiber according to the invention, in which the outer cladding is formed by air-holes placed in concentric circles, and where the inner cladding is formed by air holes of a non-circular cross section.\",\n \"a) Four glass segments are used to keep the central part of the fiber in place.\",\n \"b) Three glass segments are used to keep the central part of the fiber in place.\",\n \"FIG.\",\n \"38 shows schematically the effective index variation of microstructures that may be utilised for realising fibers according to the present invention with a core comprising material with a maximum refractive index being lower than a highest refractive index of any material in the remainder of the fiber.\",\n \"FIG.\",\n \"39 schematically shows the cross-section of a micro-structured photonic band gap fiber according to the invention.\",\n \"A honeycomb-structured cladding has been used, and the core is formed by an additional hole.\",\n \"Surrounding the central hole is a doped region.\",\n \"FIG.\",\n \"40 schematically shows the cross-section of a micro-structured photonic band gap fibre according to the invention.\",\n \"The fiber contains an annular ring with a number of additional air holes placed to enhance the dispersion compensating properties of the PBG fiber, and the outer core comprises small low-index features.\",\n \"FIG.\",\n \"41 illustrates the effective index of a band gap guided mode in a fiber structure according to the invention.\",\n \"FIG.\",\n \"42 shows a simulation of the inverse group velocity dispersion that may be obtained for fibers according to the present invention.\",\n \"FIG.\",\n \"43 shows microscope pictures of a real fiber according to the present invention.\",\n \"The figure further shows the dispersion characteristics of the fiber at wavelengths around 1550 nm.\",\n \"FIGS.\",\n \"44a-44c show microscope pictures of another real fiber according to the present invention.\",\n \"The figure further shows the dispersion characteristics of the fiber at wavelengths around 1550 nm.\",\n \"FIGS.\",\n \"45a-45c show another example of a real fiber according to the present invention.\",\n \"FIGS.\",\n \"46a-46b provide a schematic example of a fiber according to the present invention.\",\n \"FIGS.\",\n \"47a-47b provide a similar schematic example.\",\n \"4.DETAILED DESCRIPTION In FIG.\",\n \"1a, the index profile of standard step-index fiber design according to prior-art is illustrated.\",\n \"In this type of fiber design, the core region 101 is generally formed by a narrow circular section, which is strongly co-doped by germanium to provide the raised index in comparison with the surrounding cladding region 102.It should, however, be noted that the fiber also may be realised by an un-doped (pure silica) core surrounded by a doped cladding region (typically F-doped silica).\",\n \"The design also includes a thin solid rod of glass placed in air (or vacuum) or even another lower index material compared to the core glass.\",\n \"In FIG.\",\n \"1b, the equivalent microstructured fiber is shown schematically.\",\n \"In this realisation, an un-doped core 103 is surrounded by a cladding 104 that is formed by placing air holes 105- or voids—in a silica background material.\",\n \"This realisation of the cladding provides an average refractive cladding index, which is lower than the refractive index of the core.\",\n \"The inscribed, dashed circle 106 in FIG.\",\n \"1b indicates one division between the core region and the cladding region—defining the core diameter, Dco, as the inscribed diameter.\",\n \"In FIG.\",\n \"2a, the refractive index profile of a prior art fiber of the so-called w-profile type is illustrated.\",\n \"This type of refractive index is characterised by a core 201—typically formed by an index raised by germanium co-doping.\",\n \"The core is surrounded by a depressed (typically through flour co-doping) cladding region 202, and this inner cladding region is surrounded by an outer cladding 203, which typically is realised in pure silica.\",\n \"As described in the background description of the invention, these fibers operate by a spectrally dependent effective mode index, which is related to spectrally dependent power redistribution of the fundamental mode.\",\n \"FIG.\",\n \"2b (prior art) illustrates an example of a micro-structured fiber realised to form an equivalent to the W-profile fiber of standard technology.\",\n \"Here, the core is formed by a solid core region 204 surrounded by an inner cladding region 205 in which air holes 206 is placed.\",\n \"Since the air holes 206 of the inner cladding region have a larger cross section than the air holes 208 of the outer cladding 207, the effective index of the inner cladding becomes smaller than that of the outer cladding.\",\n \"Generally, this type of W-profile fiber will show significant macro-bending losses near the wavelength where a significant negative dispersion is obtained.\",\n \"In FIG.\",\n \"3a (prior art), the index profile of a triple-cladding standard optical fiber for dispersion compensation (DCF) is illustrated.\",\n \"A DCF of this type has a narrow highly-doped core 301 (typically germanium is used as index raising dopant) surrounded by a depressed cladding 302 (typically flour-doped to refractive index levels in the order of −0.5%) This germanium doping in the central part of the fiber may typically result in an increase of the refractive index of 2-3% using rather narrow core radii in the order of 1-2 μm.\",\n \"The highly doped core is favourable as it enables a high negative dispersion, but the drawback is increased attenuation (as described by Grüner-Nielsen et al., ECOC'2000, pp.91-94).\",\n \"The depressed cladding decreases the dispersion and is necessary to obtain negative dispersion slope around 1550 nm.\",\n \"Another index ring 303 with raised index compared to the cladding is often introduced to improve bending properties, but the refractive index is normally not raised much above 0.5%, since a high index value will lead to multimode operation of the fiber.\",\n \"Outermost the fiber has a pure silica cladding 304.FIG.\",\n \"3b (prior art) illustrates the transversal cross section of a microstructured optical fiber designed to make use of similar (but enhanced) properties compared to standard fiber technology.\",\n \"The illustration shows the core 305 formed by a solid material.\",\n \"The core is surrounded by an inner cladding 306 in which air holes having relatively large cross sections 307 are placed to form an effective depressed cladding.\",\n \"The inner cladding is, furthermore, surrounded by a second region 308 in which small air holes 309 are placed to form a higher effective index compared to that of the inner cladding region 306.Finally, the waveguide consist of an outermost cladding 310 in which air holes 311 of a medium size (compared to those of the inner regions) are placed.\",\n \"It should be noted that we throughout this description generally will assume that the basis material is silica or doped silica, since this is the most commonly used material for fabrication of optical fibers.\",\n \"However, the principles and ideas behind the invention is not limited to these material combinations, and it may for future applications, where new spectral ranges of the optical fiber technology is explored, be more advantageous to use different material compositions, such as (but not limited to) different compound glasses (e.g.\",\n \"chalcogenide), polymers and/or low-melting point glasses.\",\n \"Microstructured fibers are generally divided into two types depending on the physical mechanism responsible for the light guidance: index-guiding microstructured fibers are characterised by a core having a higher refractive index than a surrounding cladding region (as known from conventional optical fibers), and photonic band gap micro-structured fibres are characterised by a core having effectively a lower refractive index than a surrounding cladding region having periodically distributed features (see Broeng et al, Optical Fiber Technology, Vol.\",\n \"5, pp.\",\n \"305-330, 1999 for a presentation of the classification of the two groups).\",\n \"While the physical mechanism causing the waveguidance in the two types of microstructured fibers is different, the improvements disclosed by the present inventors relate to both types of microstructured fibers.\",\n \"In FIG.\",\n \"4 (prior art) is depicted how the effective index of the fundamental space filling mode (the “effective refractive index”), β/k, of a periodic structure varies with the normalised wavelength, λ/Λ.\",\n \"Here λ is the free space wavelength of light, Λ is the distance between the centres of two nearest neighbouring air holes, β is the propagation constant and k is the free space wave-number, all of which are well known by those skilled in the art.\",\n \"The periodic structure consists of circular air holes (the refractive index of air is 1.0 in the calculations) placed on a triangular lattice on a silica background (the refractive index of silica has been approximated by a refractive index of 1.45).\",\n \"Three curves are shown: the curve at the top 401 is for a periodic structure where the air holes are placed in a triangular lattice (also known as close-packed arrangement) and have a diameter, d, of 0.2Λ, where Λ is the lattice period (equal to the center-to-center of the air holes for the triangular lattice).\",\n \"The curve 402 is for a similar structure but with d=0.3Λ, while the curve at the bottom 403 is for a similar structure but with d=0.5Λ.\",\n \"Notice that the effective refractive index is low and not strongly dependent on the wavelength when the wavelength, λ, is large compared to the structural size (symbolised by Λ).\",\n \"It is known from the prior art that a low refractive index of the cladding is an advantage if strong dispersion compensation is desired.\",\n \"If a photonic crystal fiber (PCF) with a silica core region (see FIG.\",\n \"1b), surrounded by a periodic cladding region (such as the ones for which effective indices have been calculated as shown in FIG.\",\n \"4) is to be used as a dispersion compensating fiber one may, therefore, assume that an inner cladding region with a small pitch, compared to the wavelength, is to be used, since this gives the lowest cladding index according to the effective index curves for periodic structures shown in FIG.\",\n \"4.Furthermore, since the effective index of the cladding according to FIG.\",\n \"4 is lowest when large air filling fractions are employed, one may assume that one should employ cladding structures with large air-filling fractions to obtain maximum dispersion compensation.\",\n \"This is in agreement with the teachings by Birks et al.\",\n \"in IEEE Photonics Technology Letters, Vol.\",\n \"11, pp.674-76, 1999 and Optical Fiber Communication Conference, FG2-1, pp.\",\n \"108-10.1999, and DiGiovanni et al.\",\n \"in U.S. Pat.\",\n \"No.\",\n \"5,802,236.In the Birks-references, the cladding index is assumed to be one (which is of course less than the actual index of any real life microstructured cladding), while the core is supposed to be a circular glass-rod of very small diameter (0.4-1.2 μm).\",\n \"According to the invention, PCFs for dispersion compensating purposes can, therefore, be made with a cross-section similar to the one shown schematically in FIG.\",\n \"1b.\",\n \"If the air-filling fraction of the cladding is large, the group velocity dispersion will typically have minimum values when the inter-hole distance of the cladding is small compared to the wavelength.\",\n \"This corresponds to a core diameter, which is small compared to the wavelength, in agreement with the findings by Birks et al.\",\n \"One should notice that a number of problems are associated with small core sizes: coupling light into the fiber using standard splicing techniques is associated with significant coupling losses because of a poor mode-overlap with the standard fiber coupled from or to.\",\n \"This is well understood by those skilled in the art.\",\n \"Further, small core sizes enhance non-linear effects in the high bit rate systems of the future.\",\n \"This is also well understood by those skilled in the art.\",\n \"However, the group velocity dispersion values that may be obtained, using the schematic design shown in FIG.\",\n \"1b, are very impressive: group velocity dispersion may be as low as −2000 ps/nm/km according to Birks et al.\",\n \"It is a disadvantage of the microstructured fibers with a cross-section similar to the one depicted in FIG.\",\n \"1b with group velocity dispersion less than −400 ps/nm/km, that the fibers have small core size of around or less than 1 μm and, therefore, poor coupling coefficient to standard optical fibers.\",\n \"It is a further disadvantage of the microstructured fibers with a cross-section similar to the one depicted in FIG.\",\n \"1b with group velocity dispersion less than −400 ps/nm/km, that the fibers have small effective mode-areas and, therefore, suffer from non-linear effects in the future high bit rate transmission system.\",\n \"Such fibers would, therefore, have a detrimental effect on the bit rates that can be transmitted by the future multi-gigabit systems.\",\n \"The present inventors have realised that a number of ways exist of designing microstructured fibers with large negative group velocity dispersion, while avoiding the small core sizes that results from the large index contrast inherent in the designs suggested so far for dispersion compensating PCFs.\",\n \"To understand the basic idea, consider again the mode-indices of FIG.\",\n \"4.If one could design a fiber with a core index that abruptly rises above the cladding index as the frequency is increased, one would obtain a mode-index that curves upward in an index plot similar to the index plot shown in FIG.\",\n \"4.This corresponds to large negative group velocity dispersion, GVD, according to the formula for group velocity dispersion: GVD = - λ c ⅆ 2 n ⅆ λ 2 .\",\n \"The only apparent problem with this approach is that the index curves shown in FIG.\",\n \"4 never seem to cross.\",\n \"However, the index curves shown in FIG.\",\n \"4 are for a specific class of periodic structures.\",\n \"The problem has already been solved for microstructured fibers of the prior art: By enlarging the inner cladding air-holes, such as shown schematically on FIG.\",\n \"2b, the field is not allowed to be guided in the core region with a mode-index, which exceeds that of the cladding index at large wavelengths.\",\n \"At some wavelength, the mode-index of the fundamental cladding mode rises very abruptly, leading to large negative group velocity dispersion over a limited wavelength range.\",\n \"This is because the fundamental mode is localised within the central core region at short wavelengths, which causes the mode-index to rise abruptly, while the mode at the same time becomes guided.\",\n \"Unfortunately, this abrupt rise in mode-index happens, when the core region is relatively small compared to the core region of standard optical fibers, thus giving the same problems with non-linear effects and poor coupling as the basic fiber design depicted in FIG.\",\n \"1a.\",\n \"The present inventors have realized, however, that by doping the central core region, giving it an index of refraction that is higher than the index of refraction of the background material of the cladding region, one may obtain large negative group velocity dispersion, while having larger core regions than the core regions of the prior art fibers.\",\n \"Such a design is shown schematically in FIG.\",\n \"5, which shows the higher refractive index of the core region 500, the inner cladding air holes 501, the outer cladding air holes 502 and the background material 503 with a lower refractive index than 500.It should be noticed that the doped core is not in contact with the inner cladding features 501.As shall be discussed in more details later, in connection with FIG.\",\n \"31, the present inventors have realized that this provides an advantageous design direction for lowering the mode field overlap with the low-index features and thereby lowering potential loss-mechanisms associated with the low-index features and their interfaces with the background material of the fiber (such as absorption and scattering).\",\n \"In FIG.\",\n \"6 (prior art) there is illustrated a number of mode-indices of periodic structures, which illustrates why effective dispersion compensation may be obtained by a fiber design as illustrated in FIG.\",\n \"5.The curve at the top 601 repeats the effective index of the curve 401 of FIG.\",\n \"4—this curve is referred to as the outer cladding index, since it is supposed to represent the outer cladding structure of the micro-structured fiber shown in FIG.\",\n \"5 (502 and 503).\",\n \"The illustration also shows the effective indices of hypothetical honeycomb periodic structures, where the background has an index of refraction of 1.46.In a sense, these honeycomb structures may represent a periodic version of the central core region and its surrounding 6 holes of inner cladding structure—see for example Riishede et al.\",\n \"Proc.\",\n \"27th Eur.\",\n \"Conf.\",\n \"On Opt.\",\n \"Comm (ECOC'01), pp.\",\n \"Th.A.1.5, 2001.Three curves are shown: curve 602 corresponds to a honeycomb-structure with a hole diameter, d, of 50% of the inter-hole distance, Λ, (center-to-center), 603 corresponds to d=0.6Λ, and 604 corresponds to d=0.7Λ.\",\n \"The present inventors have realized that any strictly guided modes must have an effective index that lies above the effective cladding index, yet below the effective refractive index of the core region (approximated by the honeycomb-index).\",\n \"At some wavelength (λ/Λ around 0.5-0.6 for the design parameters of this example), the honeycomb indices are seen from FIG.\",\n \"6 to rise above that of the outer cladding index.\",\n \"This is caused by the higher refractive index of the central core region compared to the background refractive index of the cladding region.\",\n \"The central core, therefore, only guides light at shorter wavelengths than the wavelength, where the honeycomb index and the cladding index cross (to the left of the crossing point in FIG.\",\n \"6).\",\n \"As before, the guiding is associated with the fundamental mode being localised to the core region, thus giving large negative group velocity dispersion, when the cladding index abruptly rises above the core index—here the mode field distribution of the fundamental mode will shift away from the central core.\",\n \"However, by doping the central core we can employ small holes—or complete elimination of these—in the outer cladding structure, and quite large holes in the inner cladding region, and still obtain a crossing wavelength.\",\n \"Actually, a crossing wavelength will always exist, if the core region has a region with higher refractive index than any index of refraction in the outer cladding region.\",\n \"One may vary the hole-size and obtain different crossing points.\",\n \"FIG.\",\n \"6 also illustrates that enlarging the inner cladding air holes moves the crossing point to the left.\",\n \"This is an advantage, since for a fixed operational wavelength, λ, this corresponds to a larger structure size, Λ, and, therefore, a larger core region.\",\n \"To exemplify; 602 crosses 601 at λ/Λ=0.57, 603 crosses 601 at λ/Λ=0.46 and 604 crosses 601 at λ/Λ=0.44.Choosing λ=1.55 μm as the operation wavelength this corresponds to Λ=2.72 μm for 602, Λ=3.37 μm for 603 and Λ=3.52 μm for 604.One can, therefore, adjust the core size corresponding to large negative group velocity dispersion relatively freely, by simply adjusting the inner cladding hole size.\",\n \"The above-mentioned Λ-values correspond to inscribed core diameters of between: 4.1 μm for 602, 4.7 μm for 603, and 4.6 μm for 604.Other possibilities of adjusting the core size to a desired value exist, however.\",\n \"The cladding index 601 is repeated as 701, and the honeycomb index for a fiber with an inner cladding-hole diameter of 60% of the inter-hole distance 603 is repeated as 703 in FIG.\",\n \"7.The diameter of the core region with refractive index of 1.46 is equal to the inter-hole distance in 703.In FIG.\",\n \"7 is also shown the dramatic effect of changing the doping levels of the central core region: If the refractive index of the region with increased index is raised to 1.47 (curve 702) while maintaining the diameter (relative to the hole distance) of the doped region, the crossing point may move significantly to the right (from λ/Λ=0.46 to λ/Λ=0.72).\",\n \"This can be counter-acted by reducing the diameter of the doped region to 50% of the inter-hole distance as shown by 704.For a wavelength of 1.55 μm (curve 702) corresponds to Λ=2.15 μm, curve 703 corresponds to Λ=3.37 μm and curve 704 corresponds to Λ=3.52 μm at their respective crossings with curve 701.A large number of possibilities, therefore, exist for adjusting the core size (and thereby the mode field diameter) with this design.\",\n \"These possibilities include: 1) The value of the cladding index.\",\n \"The higher the cladding index the larger the core for a given core design.\",\n \"The present inventors have realised that largest core region is obtained when the outer cladding is a homogeneous material (such as pure, fused silica or silica glass doped with for example Fluorine).\",\n \"2) The size of the inner cladding air holes.\",\n \"3) The maximum of the refractive index in the doped core and the refractive index profile.\",\n \"Generally, a larger refractive index step between the central core and the background material of the outer cladding leads to a smaller core size as shown by FIG.\",\n \"7.4) The diameter of the up-doped central part of the core region.\",\n \"Generally a smaller diameter leads to larger Λ, and thereby a larger core area, and larger mode field diameter, as shown by FIG.\",\n \"7.The actual combination of parameters that should be employed depends on the specific application of the micro-structured fiber.\",\n \"For dispersion and dispersion slope compensating applications, there is a general interest in core regions that are relatively large in order to eliminate degrading effects caused by non-linearity.\",\n \"The present invention, therefore, applies to fibers with a core diameter that is more than three times the free space wavelength of the light that should be dispersion compensated.\",\n \"For standard purposes (dispersion compensation in the so-called communication window) this means that that the core diameter of the fibers of the present invention should be larger than 4.0 μm.\",\n \"On the other hand, the design ideas outlined in the present invention may also be used to tailor the dispersion to zero or close-to-zero at wavelength around 1550 nm for fibers with small core sizes—such as core sizes below 4 μm in diameter.\",\n \"The present invention, therefore, also covers fibers for non-linear applications.\",\n \"The present invention does not limit itself to structures with the principal design shown in FIG.\",\n \"5.It is well known that dispersion-compensating fibers with a depressed cladding region tend to have large bending losses.\",\n \"As a general rule, the bending losses increase when a larger core region is employed.\",\n \"Since the present invention in a main aspect relates to large core regions it would be advantageous to design the fiber such that bending losses are suppressed.\",\n \"The present inventors have realised that this is indeed possible with the type of design shown schematically in FIG.\",\n \"8.The core region 800, the inner cladding region comprising low-index features 801, and the outer cladding region 803 and 804 are similar to those shown in FIG.\",\n \"5.However, some of the innermost cladding air holes have been removed at positions 802.This effectively creates what may be termed a raised outer cladding region (similar to that shown in FIG.\",\n \"3 and denoted 308), or alternatively it may be termed an outer core region 802 if one follows the line of thought presented by Thyagarajan et. al.\",\n \"in IEEE Photonics Technology Letters, Vol.\",\n \"8, no.\",\n \"11, November 1996, pp.\",\n \"1510-12.Note, however, that compared to the approach by Thyagarajan et al., the microstructured fiber does not need high doping levels, which are difficult to control, and we may even operate with much larger effective index contrasts.\",\n \"Since one may in some sense view the inner core 800 and the inner cladding 801 as one core (which was theoretically exploited to let honeycomb indices represent the core index in FIG.\",\n \"6), we approximately know the slope of the effective guided mode index of the central core region.\",\n \"Looking at FIG.\",\n \"6, we realise that strong negative group velocity dispersion will result, if the slope of this mode is numerically large, while the slope of a mode guided mainly within an outer core is numerically small.\",\n \"In designs such as that shown in FIG.\",\n \"3a, the negative group velocity dispersion depends critically on the type of design.\",\n \"If 303 cannot be seen as capable of supporting guided mode(s), only modest dispersion may result (a typical result will be less than nominally 300 ps/nm/km).\",\n \"If on the other hand 303 represents an outer core, strong dispersion may result (even more negative than −5000 ps/nm/km).\",\n \"It should be understood that by using the term “represent an outer core” is meant that the core may support a guided mode, but not necessarily does so—for example if no light is coupled into the core from the input end of the fiber.\",\n \"With designs such as that shown in FIG.\",\n \"8 on the other hand, the numerical value of the dispersion may be large in both cases (and actually can be equally large in structures such as the one shown in FIG.\",\n \"5).\",\n \"This is because the core index may have a large negative slope, when it crosses the cladding index, as illustrated in FIG.\",\n \"6.One additional functionality is that the numerical value of the group velocity dispersion of micro-structured fibers such as shown in FIG.\",\n \"8, therefore, does not depend critically on whether the outer core guides an independent mode in region 802 or not—it may still affect the guided mode in the central core 800.It may still be advantageous to have an outer core capable of supporting a mode.\",\n \"This is because this will lift the effective index of the fundamental mode above the effective cladding index at the wavelength regime of strong dispersion and thereby reduces the bending losses.\",\n \"FIG.\",\n \"9 shows the operation of a microstructured fiber with the design schematically shown in FIG.\",\n \"8.Curve 901 shows the effective core index as a function of the normalised wavelength, curve 903 shows the effective cladding index as a function of the normalised wavelength and curve 902 shows the effective mode-index of the fundamental mode as a function of the normalised wavelength.\",\n \"For this fiber, the refractive index of the up-doped central part of the inner core 800 is 1.46.The diameter of 800 is equal to the inter-hole distance, Λ.\",\n \"The refractive index of the background material of the rest of the fiber 804 is 1.45.The diameter of the inner cladding air holes is 0.6Λ.\",\n \"The diameter of the outer cladding air holes is 0.2Λ.\",\n \"Six of the innermost outer cladding holes were removed to create the outer core region 802 in the manner shown in FIG.\",\n \"8.Looking at FIG.\",\n \"9, it is seen how the mode-index of the fundamental mode 902 has a slope similar to the mode-index of the cladding 903 at long normalised wavelengths.\",\n \"At normalised wavelengths near 0.45 the slope of the fundamental mode 902 changes and approaches the slope of the core index 901.This gives a strong upward curvature on 902 that correspond to numerically large negative group velocity dispersion of the fundamental mode 902.In this wavelength regime, where the fiber exhibits strong negative dispersion, the central and the outer core may support modes of approximately similar effective index.\",\n \"The actual group velocity dispersion is shown in FIG.\",\n \"10 for a fiber such as the one described above for FIG.\",\n \"9.FIG.\",\n \"10 is for a fiber with Λ=3.45 μm.\",\n \"This corresponds to a central core 800 with refractive index=1.46 and diameter=3.45 μm.\",\n \"The inner cladding air holes have diameter 2.07 μm (giving an inscribed diameter of 4.8 μm), while the outer cladding air holes have diameter 0.69 μm.\",\n \"The background refractive index is 1.45.In agreement with the effective index variation 902, FIG.\",\n \"10 shows strong negative group velocity dispersion, GVD, of less than −1600 ps/nm/km.\",\n \"Designs with larger doping levels show even numerically larger group velocity dispersion values (down to −10000 ps/nm/km).\",\n \"Also the large negative slope on the group velocity dispersion at wavelengths shorter than 1.55 μm should be noticed.\",\n \"Such a large dispersion slope makes these fibers ideal for dispersion slope compensation schemes.\",\n \"In some instances, the slope of these dispersion compensation fibers may even be too large.\",\n \"However, systems having smaller slope (relative to the GVD-values) may be created by cascading fibers with different GVD characteristics.\",\n \"It is, therefore, an advantage of these fibers that they may be used to not only compensate the dispersion but also the dispersion slope of virtually any system with positive group velocity dispersion.\",\n \"In order to illustrate the good mode confinement of the dispersion compensating fiber designs according to the present invention, FIG.\",\n \"11 shows the mode field distribution of the fundamental mode calculated at a wavelength of 1550 nm.\",\n \"The fiber parameters are selected according to the description provided in connection with the illustration of the group velocity dispersion in FIG.\",\n \"10.The field is seen to avoid the air holes in the inner cladding region, being advantageous in terms of low losses—as previously discussed.\",\n \"This being mainly attributed to the core region having a high-index features being not in contact with the inner-cladding region.\",\n \"In a preferred embodiment of the present invention, the outer core contains material with a higher refractive index than the refractive index of the background material in the outer cladding.\",\n \"An example of a fiber that has a number of high-index features present in the outer core is illustrated in FIG.\",\n \"12.In order to reduce fabrication efforts, it may further be preferred to eliminate the use of low-index features in the outer cladding.\",\n \"The fiber in FIG.\",\n \"12, therefore, consists of a first high-index type core region 1200, a single layer of low-index features 1201 surrounding the core and a single layer of high-index features 1202 surrounding both the core region 1200 and the low-index features 1201.Outside the core region only a single background material 1203 is present—this background material being similar for the inner cladding region, the outer core region and the outer cladding region.\",\n \"Due to the higher refractive index of the features 1202 compared to the background material and the low-index features 1201, the ensemble of high-index features may act as a core in its own right that may support a propagating mode.\",\n \"The outer core may also be formed from an annular region 1204 of high refractive index material.\",\n \"The two core regions (these being the central core region 1200 and the ensemble of high-index features 1202 or the annular region 1204) thereby are able to support each their different core modes.\",\n \"These core modes will be decoupled at most wavelengths and only couple to each other over a relatively short wavelength range.\",\n \"Within this short wavelength range, the mode distributions in the fiber will change dramatically and the fiber will exhibit a strong dispersion.\",\n \"The qualitative operation of this type of fiber is schematically illustrated in FIG.\",\n \"13.As a function of wavelength, FIG.\",\n \"13a shows schematically, the effective index of a mode that is guided in a fiber comprising only the central core region 1200 and of a mode that is guided in a fiber comprising only the outer core 1201 (this outer core mode being supported by the ensemble of high-index features 1203 or the annular region 1204).\",\n \"The two mode curves are seen to cross at a normalised wavelength, λcoupling/Λ.\",\n \"Around this wavelength, the two cores will couple strongly to each other for a composite fiber comprising both cores.\",\n \"At the coupling wavelength, for a fiber with a design as for example shown in FIG.\",\n \"12, the effective indices of modes supported by the two cores will have a lifted degeneracy—or so-called avoided crossing will take place.\",\n \"In this manner the fiber is capable of guiding light in a fundamental mode corresponding to the curve 1303 that exhibits a strong negative dispersion at wavelengths around λcoupling/Λ.\",\n \"The fiber may support a second mode guided indicated by the curve 1304, but it is not necessary that the fiber carries any substantial energy in the mode 1304 in order to exhibit negative dispersion in mode 1303.The fiber of FIG.\",\n \"13b will guide light in the central core at wavelengths shorter than the wavelength of strongest coupling (roughly equal to the coupling wavelength indicated in FIG.\",\n \"13) and the fiber will here exhibit negative dispersion with a negative dispersion slope—this is the range that dispersion compensating fibers typically are preferred to operate in.\",\n \"The wavelength-axis in FIG.\",\n \"13 has been normalised with respect to pitch (for example the center-to-center spacing between two adjacent low-index features 1201) to indicate that the coupling wavelength may be changed by changing the pitch.\",\n \"The figure also indicates the index of a homogeneous cladding with a refractive index of around 1.444—corresponding to pure silica being used as background material 1203.The higher refractive index of the core 1200 as well as of the high-index features 1202 compared to the background material 1203 cause the coupling wavelength to occur in a regime where both the index of both core modes are above the cladding index.\",\n \"In this manner, the coupling—and thereby the strong dispersion—may be obtain while the fiber is not sensitive to bending losses.\",\n \"It is worth noticing that the mode indices of both core modes may be lower than the cladding index at long wavelengths.\",\n \"This is due to the low-index features 1201 having a refractive index significantly below that of the cladding.\",\n \"The core modes may, however, well be guided for mode indices below the cladding index—as is the case for conventional one-material photonic crystal fibers—but the modes may in this case experience leakage losses.\",\n \"These leakage losses are typically reduced for one material photonic crystal fibers by having a sufficient number of low-index features surrounding the core region that supports a propagating mode—and the outer cladding may be micro-structured as for the fiber in FIG.\",\n \"8.For homogeneous outer cladding regions, the potential problem of leakage-loss at the coupling wavelength may be eliminated by the use of features 1202 having a higher refractive index than the background material 1203.It is an advantage that there are different maximum refractive indices in the two core regions—and that the core region with the lower maximum refractive index has a larger special extend than the other core region.\",\n \"As an example, the inner or central core region (referred to as first core) may have a maximum refractive index of 1.47 and the outer core region (referred to as second core) may have a maximum refractive index of 1.465.The first core should then be designed to have the smallest cross-sectional area of the two cores, whereby the index curve of the mode guided in the first core will have a more steep slope than the index curve for the mode supported by the second core.\",\n \"The magnitude of the negative group velocity dispersion depends on various things.\",\n \"The strength of the coupling between the two modes may be quantified by the field overlap.\",\n \"At λcoupling/Λ (where the two un-coupled modes cross) the splitting of the two modes will be proportional to the coupling strength.\",\n \"Often the coupling strength will only have weak wavelength dependence and in that case the wavelength-range where coupling is efficient in changing the modes is strongly related to the slopes of the two mode curves.\",\n \"When the slopes of the coupling mode curves are as different as possible, the effective wavelength-range will be small and the modes (which have a curved behaviour) will get the smallest possible bending curvature, i.e.\",\n \"numerically large group velocity dispersion will be exhibited when the slopes are as different as possible.\",\n \"The present inventors have realized that microstructured fibers with a coupled-core design may also be advantageous for non-linear applications, where it is desired to have fibers with small core and a dispersion close to zero over a broad wavelength range (e.g.\",\n \"around 1550 nm)—such as in the wavelength range from 1300 nm to 1700 nm (this is still in the regime of negative waveguide dispersion).\",\n \"Looking at FIG.\",\n \"13, it is seen that it will be preferred to move the coupling wavelength to smaller values of wavelength/pitch in order to reduce the fiber dimensions at a given wavelength.\",\n \"Hence, it may be preferred that the two core regions have a maximum refractive index that is close to each other and that the area of the two core regions is not too different.\",\n \"Alternatively, it is preferred that the outer core region has an effective refractive index close to the (effective) refractive index of the cladding.\",\n \"According to the present invention, a number of characteristics are preferred for fibers as schematically shown in FIG.\",\n \"12: Core region 1200 having a maximum refractive index, Nco,1.Typically, silica doped with Germanium and/or other doping materials will be used to form the core.\",\n \"Various refractive index profiles of the core may be preferred, but typically a profile as close to a step-index profile will be preferred.\",\n \"Hence, typically Nco,1 is larger than 1.444, such as preferably larger than 1.46, such as larger than 1.47 at wavelengths around 1550 nm.\",\n \"Background material 1204 having a refractive index, Ncl,2, that is lower than Nco,1.Typically, the background material is pure silica, having a refractive index of around 1.444 at wavelength around 1550 nm, but silica doped with a doping material to lower the refractive index of Ncl,1 below 1.444 may also be preferred, such as fluorine-doped silica glass.\",\n \"Low-index features 1201 having a refractive index, Ncl,1, a diameter, dcl,1, and a center-to-center spacing between two adjacent low-index features, Λcl,1.Typically, these low-index features are voids, hence Ncl,1 is around 1.0, but low index features formed from down-doped silica may also be preferred.\",\n \"For dispersion compensation applications, typically the low-index features will have a diameter, dcl,1, in the range from around 0.5Λcl,1 to 0.9Λcl,1 and for non-linear fiber applications, typically, dcl,1 will be in the range from 0.2Λcl,1 to 0.5Λcl,1.In order to reduce degrading effects, such as polarisation mode dispersion, PMD, it is often preferred that all low-index features are identical and equally spaced.\",\n \"Also other numbers of low-index features than 6 is covered by the present invention as well as more than one “ring” of low-index features surrounding the core region 1200.High-index features 1202 having a refractive index, Nco,2, a diameter, dco,2, and a center-to-center spacing between two adjacent high-index features, Λco,2.Typically, these high-index features comprise doped silica glass, hence Nco,2 is larger than 1.444 at wavelengths around 1550 nm, but high-index features formed from pure or down-doped silica may also be preferred.\",\n \"For dispersion compensation applications, typically the high-index features will have Nco,2 in the range from 1.444 to 1.460 at wavelengths around 1550 nm and a diameter, dco,2, in the range from around 0.1Λco,2 to 0.9Λco,2 and for non-linear fiber applications, typically, dco,2 will be in the same range.\",\n \"Furthermore, in order for the ensemble of high-index features to support only a single-mode, it is often preferred that the size of the high-index features is smaller than the size of the low-index features 1201.The present inventors have further found that it is an advantage that the refractive index profile of the high-index features is as close as possible to a step-index profile in order for the coupling to be as dramatic as possible—and thereby the dispersion as strong as possible.\",\n \"In order to reduce degrading effects, such as polarisation mode dispersion, PMD, it is often preferred that all high-index features are identical and equally spaced.\",\n \"Alternatively, the ensemble of high-index features in the outer core may be substituted by a homogeneous high-index annular region 1204.Such an annular region—being either the complete or parts of the outer core region—may have the same functionality as the ensemble of high-index features.\",\n \"The width, the maximum refractive index and the index profile of the annular region are key parameters for the fiber.\",\n \"Also other numbers of high-index features than 6 is covered by the present invention as well as more than one “ring” of low-index features surrounding the core region 1200.In order to optimise the coupling between modes supported by the first core region 1200 and the second core region (formed from the high-index features 1202), it is often preferred that the high-index features are positioned such that light penetrating between two low-index features from the core region 1200 will experience the high-index features most strongly.\",\n \"To quantify this, it may be preferred that a first polygon 1400 drawn from centres of high-index features and a second polygon 1401 drawn from centres of low-index features are concentric, but rotated with respect to each other an angle given by 0.5(N1/360′), where N1 is the number of low-index features.\",\n \"Hence, for a fiber with 6 low-index features (as in FIG.\",\n \"14), the first polygon 1400 is rotated 30 degrees compared to the second polygon 1401, and both polygons are coinciding.\",\n \"Alternatively, the second polygon may have a similar orientation as the first polygon.\",\n \"As schematically illustrated in FIG.\",\n \"15, it may also be preferred that the outer cladding region 1501 comprises a different material—or a material with a different refractive index—than the background material 1500 of the inner cladding and/or the outer core.\",\n \"It is important to notice that the above-described design ideas for providing microstructured optical fibers for dispersion and dispersion slope compensation as well as for non-linear applications is profoundly different from the ideas presented by DiGiovanni et al.\",\n \"in U.S. Pat.\",\n \"No.\",\n \"5,802,236, where a second core region capable of supporting a propagating mode is taught to be disadvantageous.\",\n \"To illustrate in more detail the operation and some of the possibilities of fibers according to the present invention, various design parameters of fibers as shown in FIG.\",\n \"12a shall be considered next.\",\n \"FIG.\",\n \"16 shows a simulation of a fiber according to the present invention having a design as schematically shown in FIG.\",\n \"12.The fiber has the following design parameters: The first core has a step-index profile with a maximum refractive index of 1.471 (corresponding to a refractive index change of 1.892% compared to pure silica having refractive index of 1.444 at λ=1550 nm) and a background material of refractive index 1.444 (pure silica).\",\n \"The inner cladding region comprises a background material of refractive index 1.444 and six voids of diameter 0.5Λ, where Λ is the center-to-centre spacing between to two nearest voids.\",\n \"The second core region comprises a background material of refractive index 1.444 and six high-index features of refractive index 1.459 (corresponding to a raised index of 1.055% compared to pure silica).\",\n \"The outer cladding region is homogeneous and comprises pure silica with a refractive index of 1.444.While the outer cladding region is homogeneous in this case, it may also comprise varies sub-regions for example for lowering bending losses.\",\n \"As seen from FIG.\",\n \"16, the fiber supports a fundamental mode 1600 that has a strongly dispersive behaviour at normalized wavelengths, λ/Λ, of around 0.36.The curve 1601 represent the effective index of a mode that may be supported by the second core and the curve 1602 represent the refractive index of the outer cladding region.\",\n \"It should be noted that it is here chosen to use a normalisation factor, Λ.\",\n \"The fibre in FIG.\",\n \"16 will have Λ of around 4.3 μm for negative dispersion operation of around λ=1550 nm.\",\n \"This provides a core size that is significantly larger than the core sizes disclosed by the previously discussed DiGiovanni et al.\",\n \"and Birks et al.\",\n \"references.\",\n \"Furthermore, as shall be demonstrated next, the negative dispersion that may be exhibited by fibers according to the present invention can be significantly larger than for the fibers disclosed in the above-mentioned DiGiovanni and Birks references.\",\n \"For tailoring fibers according to the present invention for given specifications, a wide range of parameters may be tuned.\",\n \"The following figures show examples of how various fiber characteristics (such as dispersion, RDS and core size) may be tuned by adjusting design parameters.\",\n \"The present inventors have found that the negative dispersion depends strongly on the size of the inner cladding features.\",\n \"FIG.\",\n \"17 shows a simulation of the dispersion characteristics of fibers as in FIG.\",\n \"16, but with A ranging from 4.08 μm to 4.46 μm, and inner features being voids with a size that ranges from d/Λ of around 0.30 to 0.65.As seen from the figure, the dispersion has a characteristic “dip” with a dispersion minimum (the strongest dispersion) and a certain width.\",\n \"It is worth noticing varying the size of the inner cladding features has a strong influence on the minimum dispersion values as well as the width of the “dip” whereas the wavelength of strongest negative dispersion is only varied slightly.\",\n \"As seen from the figure, negative dispersion that is significantly beyond prior art dispersion compensating fibers is feasible.\",\n \"FIG.\",\n \"18 shows the normalized wavelength of strongest dispersion, λ*/Λ, as a function of size of the inner cladding features (solid curve).\",\n \"The figure shows that by adjusting the features size, d/Λ, from around 0.25 to 0.70, λ*/Λ can be varied from around 0.34 to 0.385.For the strongest dispersion to occur at a wavelength of around 1550 nm, this corresponds to Λ varying from around 4.03 μm to 4.50 μm (dashed curve), and d ranging from around 1.0 μm to 3.2 μm.\",\n \"These Λ values provide a relatively large separation between the high-index feature in the core 1200 and the air holes 1201.The distance from the center of the fiber to a center of an air hole 1201 is similar to Λ, hence, the afore-mentioned separation is given by s1=Λ−d−dco,1.With d/Λ ranging from 0.25 to 0.70 and dco,1 ranging from 0.5Λ to 0.8Λ this provides a separation s1 ranging up to 1.2 μm.\",\n \"For other designs that are within the present invention, the separation may be larger as may be deduced from the various examples of fibers according to the present invention.\",\n \"A non-zero separation, s1, provides an important technical advantage in terms of lowering potential losses associated with the air holes—as shall be discussed in further detail at a later stage.\",\n \"Another important feature of fibers according to the present invention is the relatively large separation between the first core 1200 and the second core (ensemble of high-index features 1202 or ring-shaped element 1204).\",\n \"This separation is typically larger than Λ, such as larger than 4.0 μm as apparent from the various examples of the present application.\",\n \"Alternatively, this advantage is apparent in the relatively large diameter of the second core (the diameter being defined as the radial distance in the cross-section from the of the fiber to a center of the second core region (such as for example a center of one of the second core features 802, 1202 or a center of the ring-shaped region 1204).\",\n \"As apparent from the various examples of fibers according to the present invention, e.g.\",\n \"as illustrated in the various embodiments in the figures, this diameter is typically in the range from 10 μm to 20 μm.\",\n \"Larger diameters may be desired for fibers operating in a higher-order mode—as shall be discussed at a later stage.\",\n \"It is important to notice, that the above-described separations and diameters are significantly larger than for fibers disclosed by Fajardo et al.\",\n \"in WO 01/98819.The above-described feature is advantageous in separating the two cores for more simple in-coupling (including splicing) such that light may be coupled substantially only into the first core region.\",\n \"For splicing to a standard transmission fiber (which dispersion is desired to be compensated) it is an advantage that the second core has a diameter larger than the core diameter of the transmission fiber.\",\n \"Typical transmission fibers have a core diameter of around 8 μm, and in order to avoid coupling into the second core at a fiber splice, it is an advantage that the diamater of the second core is larger than 8 μm (preferably larger than 10 μm) such that its overlap with the core of the transmission fiber is limited or negligible when the center of the first core of the dual core fiber is aligned to the center of the core of the transmission fiber.\",\n \"Furthermore, in order to lower in-coupling losses, it is desired that the first core—and importantly the high-index feature of the first core—has a size that is approaching the size of the core of the transmission fiber nor an intermediate fiber that has been inserted between the transmission fiber and the dispersion compensating fiber to provide mode adaption.\",\n \"In the prior art, intermediate fibers having a core size smaller than the transmission fiber is often inserted between the transmission fiber and the dispersion compensating fiber in order to adapt the core sizes.\",\n \"Typically, intermediate fibers may adapt cores from around 8-9 μm in diameter to cores of down to 4 μm.\",\n \"Hence, it is desirable to realize dual core dispersion compensating fibers with a first core having a diameter of around 4 μm to 8 μm and a second core diameter of larger than 9 μm.\",\n \"As apparent from the present application, such a feature is characteristic for a number of fibers according to the present application.\",\n \"The relatively large dimensions for the here-disclosed fibers provide several important technical advantageous as discussed throughout the application apart from the facilitated in-coupling—including relatively large MFD for lowering non-linear effects, lower PMD as lower birefringence is typically found for micro-structured fibers of larger dimensions and symmetric feature arrangements, lower losses as larger dimensions provide decreased field intensity in the air holes of the first cladding region for light guided in the first core.\",\n \"One or more of these technical advantages may be obtained for fibers according to the present invention at the same time as a stronger dispersion is obtained.\",\n \"FIG.\",\n \"19 shows a simulation of the value of minimum dispersion (strongest negative dispersion) for a fiber according to the present invention having inner cladding features varying in size from d/Λ of around 0.25 to 0.70 (normalized dispersion is indicated with a solid line—right axis).\",\n \"For operation around 1550 nm, it is possible to vary the dispersion from around −400 ps/nm/km to more than −10000 ps/nm/km (dashed curve—left axis).\",\n \"As previously mentioned, such dispersion characteristics are significantly stronger than for any prior art optical fiber.\",\n \"As seen from FIG.\",\n \"17, the width of the negative dispersion “dip” depends strongly on the size of the inner cladding features.\",\n \"Generally, a larger inner features size results in stronger dispersion, but a more narrow “dip”.\",\n \"In FIG.\",\n \"20, the width, λ, of the dispersion “dip” is simulated as a function of inner cladding features size (normalized width, □λ/Λ, is shown by solid line (right axis) and for operation at λ=1550 nm, the absolute width, □□, is shown by the dashed line (left axis)).\",\n \"The width is determined as the full-width-half-maximum (FWHM) of the numerical value of the dispersion.\",\n \"As seen from the figure, for a FWHM value of more, than 30 nm, the fiber should be designed with feature size, d/Λ, of around 0.45 or less.\",\n \"FIG.\",\n \"21 shows a calculation of the RDS values and the dispersion of another fiber according to the present invention having d/Λ=0.4 and Λ=4.214 μm.\",\n \"As seen from the figure, the fiber exhibits a strong negative dispersion of more than −500 ps/nm/km as well as a high RDS value of around 0.03 nm−1 at a wavelength of 1550 nm.\",\n \"Such a fiber is well suited for dispersion and dispersion slope compensation.\",\n \"FIGS.\",\n \"22 and 23 shows similar results for fibers having d/Λ=0.5 and d/Λ=0.6.FIG.\",\n \"24 shows another example of a fiber according to the present invention.\",\n \"In this figures, the outer core is a homogeneous material.\",\n \"The figure shows the operation of the fiber, which is similar to the previously discussed fibers of FIG.\",\n \"12.It is important to notice that the fibre exhibits an abrupt cut-off both of the fundamental mode 2400 at λ/a of around 0.28, as well as of higher-order modes at λ/a around 0.1, where a is the radius of the high-index feature in the core (the first higher-order mode is shown by the line 2403).\",\n \"This behaviour is also characteristic for the other types of fibers disclosed in the present application.\",\n \"The higher-order mode with a cut-off of around λ/a=0.1 corresponds to an LP11 mode that is supported by the first core.\",\n \"This mode will exhibit a strong negative dispersion around λ/a=0.1 that may be utilized for dispersion compensating applications.\",\n \"Hence, for operation at a given wavelength, the structural dimensions of the fiber should be scale approximately 0.28/0.10=2.8 times larger for operation in the higher order mode.\",\n \"An operation around λ/a of 0.1 corresponds to a value of around 15 μm for operation at a wavelength of around 1550 nm.\",\n \"This results in a significantly larger core size compared to the core size of the previously discussed fibers operating in a fundamental mode.\",\n \"Furthermore, as the cut-off of the higher-order mode is seen to be even more abrupt for the higher order mode compared to the fundamental mode—even stronger negative dispersion may be obtained.\",\n \"For the single-mode dispersion compensating fiber, λ=1.55 μm, dictates an a-value of around 5.5 μm, and for the higher-order mode dispersion compensating fiber, an a-value of around 15 μm is dictated.\",\n \"The present invention also relates to higher-order mode dispersion compensating fibers for other fiber designs that are disclosed by the present invention—dimensions must naturally be scaled and relations between design parameter may changes for transfer of a design for single-mode dispersion compensation to higher-order mode dispersion compensation.\",\n \"FIG.\",\n \"25 shows another schematic example of a dispersion compensating fibre according to the present invention.\",\n \"The fiber is characterized by a first core region 2500 in the center of the fiber, an inner cladding region (a first cladding region) comprising low-index features 2501 and a background material 2502, a second core region 2503 and an outer cladding region 2504 (a second cladding region).\",\n \"The first core 2500 has a size being adapted such that the core makes contact with features in the inner cladding 2501—this type of core has also been discussed by Fajardo et al.\",\n \"in WO 01/98819.The first core 2500 is characterized by a refractive index being higher than the refractive index of the features 2501 as well as that of the background material 2502 of the inner cladding.\",\n \"The second core 2503 that may either be homogenous (as shown in FIG.\",\n \"25) or it may contain a number of micro-structured features (as discussed previously for example in FIG.\",\n \"13).\",\n \"The second core comprises material with a higher refractive index compared to the effective index of the inner cladding region and a lower refractive index compared to a maximum refractive index of the first core.\",\n \"In a preferred, embodiment, the background material of the inner cladding region 2502 is identical to the refractive index of the second core 2503.The material of 2502 and 2503 may in this case be identical such as pure silica or silica comprising one or more index-raising dopants, such as for example Ge.\",\n \"An important issue for dispersion compensating fibers is their bending losses.\",\n \"The present inventors have analysed a number of fibers according to the present invention and found an important design parameter related to the cross-sectional size of the second core (or the total area of high-index feature in the second core in the case of an ensemble of high-index features forming the second core).\",\n \"This parameter is important for lowering bending losses of the fibers as shall be demonstrated using the FIGS.\",\n \"26 and 27.Looking first at FIG.\",\n \"26, the fibre is characterized by a first core with a refractive index of 1.461 and the inner cladding region comprises six air holes with diameter, d. The background material of the inner cladding region is similar to the material of the outer core and it has a refractive index of 1.444 (pure silica).\",\n \"The outer cladding comprises silica with a refractive index of 1.437, this may be obtain for example by F-doping of silica.\",\n \"The fiber in FIG.\",\n \"26 has an outer radius of the inner cladding region labelled R and an outer radius of the second core labelled R1.The fiber has R1/R=2 and d/R=0.37.In FIG.\",\n \"26 is illustrated a simulation of the operation of the fiber.\",\n \"The figure illustrates the effective indices of various modes of the fiber as a function of normalized wavelength, λ/2R.\",\n \"The curve 2600 represents the fundamental mode guided in the core region, the curve 2601 represent a higher order mode that is supported by the fiber, but not necessarily carries any energy, and the curve 2602 represent the refractive index of the outer cladding region.\",\n \"The figure shows that the curve 2600 has a strongly dispersive behaviour at normalized wavelengths around 0.9.The dispersive behaviour is noticeable by the “bending” of the curve 2600.This shows that the fiber exhibits a strong negative dispersion at normalized wavelengths around 0.9—and the fiber may, therefore, in principle, be used for dispersion compensating applications.\",\n \"There is, however, an important issue to consider for this fibre.\",\n \"This issue relates to the index difference between the curves 2600 and 2602.The two curves have a relatively small difference in the wavelength regime of strongly dispersive behaviour (for this specific example the difference is around 0.001 or less at the wavelength of strongest dispersion).\",\n \"This small difference has the negative effect that the mode corresponding to curve 2600 will not be strongly confined at the strongly dispersive regime and may experience high bending losses.\",\n \"The present inventors have realized that the potential problem of bending losses may be solved by careful design of the second core—more specifically by increasing the cross-sectional area of this region—or equivalently its width.\",\n \"As shown in FIG.\",\n \"27, the present inventors have found that for R1/R above 2, such as around 4, the strongly dispersive behaviour of the curve 2700 (to be compared with 2600 in FIG.\",\n \"26), may be lifted significantly above the refractive index of the outer cladding 2702—thereby lowering the bending losses of the fiber.\",\n \"Hence, it is advantageous to have the outer diameter of the second core cladding region being more than two times larger than outer diameter of the inner cladding region (this diameter here being defined as 2R), and preferably around 4 times larger.\",\n \"Naturally, as those skilled in the art will understand, the exact position of the strongly dispersive regime depends on a number of design parameters including the refractive index of the various materials that the fiber is composed of as well as the morphology of the various microstructures and regions in the fiber (among others, these being size, shape and arrangement).\",\n \"Considering next the dispersion that may be obtained for the fiber of FIG.\",\n \"27b, FIG.\",\n \"28 shows the normalized dispersion (solid curve), where the factor 2R has been used for normalization.\",\n \"FIG.\",\n \"28 also shows the effective mode area (dashed curve) of the fundamental mode.\",\n \"As seen, the effective area is strongly increasing as the negative dispersion regime is reached (it should be noticed that the fiber should be operated at the short wavelength side of the dispersion dip).\",\n \"The large increase in effective area is attributed to the fundamental mode being coupled to the second core at wavelengths longer than the minimum dispersion wavelength.\",\n \"It is at this stage worth comparing the fiber in FIG.\",\n \"28, to one of the previously studied fibers with a core not being in contact with the inner cladding features.\",\n \"FIG.\",\n \"29a shows dispersion and effective area of the fundamental mode for a fiber with a design as shown in FIG.\",\n \"12a.\",\n \"In FIG.\",\n \"29a, a normalization factor Λ (center-to-center hole spacing in the inner cladding region) has been employed.\",\n \"For a more direct comparison using absolute values of the design parameters, the two fibers of FIGS.\",\n \"28b and 29b may be designed for dispersion compensation around λ=1550 nm.\",\n \"FIG.\",\n \"30 shows the effective area of the fundamental mode of the fiber of FIG.\",\n \"28 (dashed curve 3001) and of FIG.\",\n \"29b (solid curve 3000).\",\n \"As apparent from the figure, both fibers may exhibit an effective area of more than 15 μm2 at wavelengths around 1550 nm, and in fact more than 20 μm2 may be exhibited by both fibers.\",\n \"While this is comparable to conventional dispersion compensating fibers, it is significantly larger than for the fibers disclosed in the previously discussed DiGiovanni et al.\",\n \"and Birks et al.\",\n \"references.\",\n \"Considering next the dispersion of the fibers according to the present invention, FIG.\",\n \"31, shows that both fibers may exhibit a negative dispersion around 1550 nm that is significantly larger than what is obtained using conventional dispersion compensation fibres as well as the fibers disclosed by Hasegawa et al.\",\n \"in EP 1 118 887.Hence, the present invention provides new dispersion compensating fibers have effective areas being comparable to conventional dispersion compensating fibers, but with significantly larger negative dispersion.\",\n \"For a further comparison of fibers having a high-index core in direct contact with the inner cladding features and a high-index core being separated from the inner cladding, FIG.\",\n \"31 shows that a stronger dispersion may be obtained in the latter case.\",\n \"A further important issue is the mode field distribution in and around the core at the wavelengths of negative dispersion.\",\n \"Looking at FIGS.\",\n \"28a and 29a, it is found that the negative dispersion occurs for λ/2R of around 0.8 for the fiber in FIG.\",\n \"28b and for λ/Λof around 0.36 for the fiber in FIG.\",\n \"29.For operation of around 1550 nm, this yields 2R of around 1.9 μm and Λ of around 4.3 μm.\",\n \"Hence, the inner cladding features for the fiber in FIG.\",\n \"29b will be positioned substantially further away from the fiber center compared to the inner cladding features of the fiber in FIG.\",\n \"28.Since both fibers have approximately similar effective area at wavelengths around 1550 nm (as shown in FIG.\",\n \"30), the field overlap of the guided mode with the inner cladding features will be substantially lower for the fiber in FIG.\",\n \"29b compared to the fiber in FIG.\",\n \"28.Generally, the inner cladding features are air holes and may be desirable to limit the amount of field in the air holes in order to lower potential loss mechanism attributed to the air holes (this may for example be absorption losses from OH content in the air holes or scattering losses from surface roughness of the air holes).\",\n \"Also in the case of lowering PMD, it may be an advantage to move the air holes as far away from the mode field as possible, since the high index contrast between the air holes and the background material may result in significant polarization effects.\",\n \"For these reasons it may be preferred to have dispersion compensating fibers according to the present invention where the first core comprises a high-index features being isolated from the inner cladding features.\",\n \"Preferably, the first core feature has a diameter of around 0.7Λ or less.\",\n \"In other cases, however, it may be preferred to have as large an overlap between the mode field and the inner cladding features as possible.\",\n \"This may for example be the case for tuneable dispersion compensating fibers, where the material in the inner cladding features may be altered in one way or another (for example by changing the refractive index of the inner cladding features dynamically using non-linear optical, acoustic-optical, or thermal processes, other mechanisms could be physical pressurizing or bending of the fibers).\",\n \"This may be particularly interesting for dispersion compensating fibres according to the present invention comprising polymers in the inner cladding features.\",\n \"Numerous possibilities exist for obtaining the required dispersion compensating effect as described in the previous text.\",\n \"In the following a number of further examples of fibers according to the present invention shall be presented.\",\n \"In FIG.\",\n \"32 is schematically shown a preferred embodiment of the desired fiber.\",\n \"Here the cross-section of a micro-structured fibre for dispersion compensation is shown having a doped central core 3201 surrounded by an annular inner cladding section containing more closely placed air holes 3202 than those used to form the micro-structured outer cladding 3203.Between the inner and outer cladding regions, a higher effective index is obtained in an annular segment 3204, where air-holes have been removed (compared to the air-filling fraction of the outer cladding).\",\n \"It should be noted that the closer spacing of the holes provides the desired lowering of the effective index, but the present type of design becomes more simple from a manufacturing point of view, because it indicates how similar effects may be obtained both through control of air-hole spacing and through control of their size.\",\n \"Yet another example of a preferred embodiment of a fiber according to the invention is schematically shown in FIG.\",\n \"33.In this embodiment, the cross-section of a micro-structured fibre for dispersion compensation is realized by having a doped central core 3301 surrounded by an annular inner cladding section containing more closely placed air holes 3302 than those used to form the micro-structured outer cladding 3303.This is in accordance with the example illustrated in FIG.\",\n \"32.However, compared to previous examples, additional air holes have been removed in the outer core region 3303, hereby allowing for a higher effective refractive index.\",\n \"Comparing FIG.\",\n \"32 and FIG.\",\n \"33, we may formulate the differences as follows.\",\n \"The fibre structure presented in FIG.\",\n \"32 has an outer annular core section formed by a micro-structured section (in which the air-hole spacing is larger than that of the outer cladding), whereas the structure illustrated in FIG.\",\n \"33 has an outer core region formed by a solid material.\",\n \"It should be noted that these examples only indicate some of the design possibilities and optimal designs according to the parameters specified in this invention include the possibilities that not only size and spacing of air holes but also background doping may be used for example using high-index features or a annular element having a center that coincides with the fiber center—as previously discussed.\",\n \"The outer core region may for example contain a number of solid, high-index features 3400 as illustrated in FIG.\",\n \"34—or be a homogeneous annular region 3500 with refractive index higher than the refractive index of the outer cladding background material, as illustrated in FIG.\",\n \"35.In the examples of the invention presented so far, the microstructured outer cladding has been illustrated as close packed air holes placed in a background material.\",\n \"It should be stressed that other air-hole distributions may be used to obtain the desired optical properties.\",\n \"Examples of such placements of air holes could be in rectangular-, honeycomb-, or Kagomé structures.\",\n \"However, even further possibilities exist, and one preferred embodiment is schematically shown in FIG.\",\n \"36, where the cross-section of a microstructured fiber according to the invention is shown.\",\n \"In this preferred embodiment the outer cladding is formed by air-holes 3604 placed in concentric circles.\",\n \"The fiber has a doped central core 3601 surrounded by an annular inner cladding section containing concentrically placed air holes 3602.Between the inner and outer claddings, the second core region 3603 is shown.\",\n \"Note that in this specific example, the second core region is formed by a homogeneous material (such as pure silica or silica doped with germanium, fluorine and/or other material(s)), but it could also be realized through a microstructured section containing low- and/or high-index features.\",\n \"The air holes that are used to tailor the effective refractive index do not necessarily have to be of circular cross sections.\",\n \"In FIG.\",\n \"37, a preferred embodiment of a fiber according to the invention is schematically shown.\",\n \"FIG.\",\n \"37a illustrates the cross-section of a micro-structured fibre according to the invention, in which the outer cladding is formed by air-holes 3704 placed in concentric circles, and where the inner cladding is formed by air holes of a non-circular cross section 3702.As in the previously discussed embodiments of the invention, the fiber has a doped central core 3701 surrounded by the annular inner cladding section.\",\n \"Between the inner and outer claddings, the second core region 3703 is shown.\",\n \"FIG.\",\n \"37b shows yet another preferred embodiment having a doped central core 3705 surrounded by an annular inner cladding section containing non-circular air holes 3706.Between the inner cladding and the outer cladding 3708, the second core region 3707 is shown.\",\n \"To further illustrate this difference between standard fibers and micro-structured fibers, we move on to showing the possibility of having a mode with large negative dispersion in a design where one would expect this to be impossible using standard index analogies between micro-structured fibers and standard fibers, such as those shown in FIGS.\",\n \"1-3.We employ a central core design such as the one shown in FIG.\",\n \"8, however, we assume the doping material does create a material with a refractive index being lower than the refractive index of silica (one could dope with e.g.\",\n \"fluorine to obtain this effect).\",\n \"Surrounding the inner cladding is an outer silica core, while the outer cladding is a micro-structured cladding with a small pitch.\",\n \"According to the standard view of the core regions (such as illustrated in FIGS.\",\n \"1-3) we, therefore, have an inner core with a refractive index being lower than the refractive index of the outer core.\",\n \"According to this view, the fundamental and the second order mode will, therefore, swap place (the fundamental mode will be in the outer core at short wavelengths, and in the inner core at long wavelengths) compared to the design suggested by Thyagarajan et al.\",\n \"in IEEE Photonics Technology Letters, Vol.\",\n \"8, pp.\",\n \"1510-12, 1996.This has detrimental effects on the performance of the fiber, which is the reason why Thyagarajan et al.\",\n \"suggested the higher index in the central core.\",\n \"The present inventors have, however, realized that it may also be preferred that the central core region does not comprise the highest refractive index material of the fiber.\",\n \"FIG.\",\n \"38 illustrates the effective core index 3802 (which is the honeycomb-index corresponding to the inner core and the inner cladding) and the cladding index 3801.The core index 3802 is seen to cross the cladding index 3801 while 3802 has a large negative slope.\",\n \"This will make it possible to have a fundamental mode with the desired dispersion characteristics, provided that the silica regions of the outer core are sufficiently small to ensure that the effective mode index of the outer core is near the outer cladding effective refractive index.\",\n \"Notice, that this crossing between the core and the cladding indices is made possible, despite the low refractive index of the core region and the large holes in the inner cladding, by a small inter-hole distance in the outer cladding region, compared to the inter-hole distance in the inner cladding region (the inter-hole distance in the inner cladding region is three times the inter-hole distance in the outer cladding).\",\n \"We now turn to a different aspect of the present invention.\",\n \"Micro-structured cladding structures with a periodically varying refractive index have the potential of exhibiting photonic band gaps.\",\n \"These are frequency regions where no light transmittance is allowed in the cladding region (see e.g.\",\n \"Barkou et al., Optical Fiber Communication Conference, FG5-1, pp.\",\n \"117-119, 1999).\",\n \"By employing a periodically varying refractive index it is, therefore, possible to obtain guidance of light within low index core regions.\",\n \"It has been shown both theoretically (Broeng et.\",\n \"al., Optics Letters, Vol.\",\n \"25, pp.\",\n \"96-98, 2000) and experimentally (Cregan et.\",\n \"al., Science, Vol.\",\n \"285, pp.\",\n \"1539-1539, 1999) that this even allows guidance of light within a hollow core, a result that cannot be explained by the theories of standard optical fibers.\",\n \"Standard fibers, on the other hand, always demand that the core region has the higher refractive index, while the cladding region has the lower refractive index.\",\n \"The present inventors have realized that one can also have a guided fundamental mode with negative group velocity dispersion with effective mode-index that is below the effective refractive index of the cladding region.\",\n \"It is important to stress that such a mode cannot be guided according to standard fiber technology.\",\n \"It is well-known from the theory of photonic band gap fibres that a mode with a mode-index below the effective refractive cladding index can only be guided if the propagation constant of the guided mode lies within a photonic band gap at the particular wavelength.\",\n \"It is not known (and not at all obvious) that it is possible to make a photonic band gap fibre design where a fundamental mode with large numerical group velocity dispersion exists.\",\n \"The present inventors have realized a way to obtain this.\",\n \"As with the designs described above we need a dual concentric core arrangement to obtain such a fundamental mode.\",\n \"Also the fundamental mode should be guided mainly in the outer core region at long wavelengths and in the inner core region at shorter wavelengths.\",\n \"As with the fibers described above the fundamental mode exhibits large negative group velocity dispersion only if the slope of the mode-index as a function of the wavelength differ significantly between the two modes when phase matching between the two potential modes occur.\",\n \"This is an analogue to the case of the preceding fibers.\",\n \"The core design, however, that may obtain this feature in photonic band gap guiding fibres turn out to be quite different.\",\n \"Consider the design shown schematically in FIG.\",\n \"39.The outer cladding consists of air holes 3904 placed on a periodic lattice (a honeycomb lattice in the example shown).\",\n \"The holes are distributed within a background dielectric material 3903, such as silica.\",\n \"The enlarged air holes 3902 correspond to the inner cladding of the preceding designs.\",\n \"They act to lower the effective refractive index of the central core region at long wavelengths so that no guided modes exist in the central core region at long wavelengths.\",\n \"At shorter wavelengths a mode becomes guided within a photonic band gap.\",\n \"This may happen even without the central hole 3900 and the high-index ring 3901 being present.\",\n \"The purpose of 3900 and 3901 will be explained later.\",\n \"In the design shown in FIG.\",\n \"40 an outer, low-index core region is included.\",\n \"It is created by adding air holes 4005 within the six innermost honeycombs.\",\n \"Notice that this would not act as a core in a standard fiber, since we have effectively lowered the effective index of the outer core region.\",\n \"In FIG.\",\n \"40 the outer cladding air holes 4004, the background material 4003, the inner cladding air holes 4002 the high-index ring 4001 and the central air hole 4000 correspond to the ones shown in FIG.\",\n \"39.In FIG.\",\n \"41 is shown some effective indices that may be obtained in a fiber with the principal design shown in FIG.\",\n \"40.The effective refractive index 4100 of the outer cladding lies at the top, corresponding to the fact that no index guided modes exist in the micro-structured fiber, at the wavelengths where the fundamental mode exhibits numerically large negative group velocity dispersion (index-guided modes exist at shorter wavelengths, however, these of not shown for reasons of clarity).\",\n \"Below 4100 are the photonic band gap edges 4101 and 4102.Any guided mode that is not an index guided mode must have a mode-index between the two photonic band gap edges, since these define the intervals that are not allowed to propagate in the cladding.\",\n \"A micro-structured fiber-structure such as the one shown in FIG.\",\n \"39 will have a mode-index as shown by 4105.The purpose of the central air-hole 4000 and the ring with raised refractive index 4001 compared to the background refractive index of the outer cladding is to obtain a mode-index 4105 which traverses the photonic band gap bounded by 4101 and 4102 as steeply as possible.\",\n \"Notice that the mode-index enters the top of the photonic band gap 4101 at short wavelengths and then descends through the band gap as the wavelength is increased.\",\n \"A fiber with only the outer core region created by the added air-holes 4005 on the other hand will have a mode-index behaving as 4106.Notice that this mode-index enters the bottom of the photonic band gap 4102 at short wavelengths and then rises through the band gap as the wavelength is increased.\",\n \"The two types of cores therefore give rise to modes that traverse the photonic band gap quite differently.\",\n \"The slope of the mode-index as a function of the wavelength of the two modes is therefore quite different when they have an equal mode-index (when they are phase-matched).\",\n \"Combining the two cores, as shown in FIG.\",\n \"26 an operation with avoided-crossing is created similar to that referred to with the index-guiding designs.\",\n \"The fundamental mode has the effective index 4103, while the second mode (that need not carry any energy) has the effective index 4104.The curvature of 4103 corresponds to numerically large negative group velocity dispersion, while the curvature of 4104 corresponds to numerically large positive group velocity dispersion.\",\n \"Notice that the slope of the two modes 4103 and 4104 tend toward the slope of the two core modes 4105 and 4106 at wavelengths differing from the wavelengths with numerically large group velocity dispersion.\",\n \"In the wavelength region with numerically large group velocity dispersion the energy of the mode 4103 is moved from the inner core region 4001 to the outer core region 4005 as the wavelength is increased.\",\n \"Correspondingly, the energy of the mode 4104 is moved from the outer core region 4005 to the inner core region 4001 as the wavelength is increased.\",\n \"The inventors have realized that a number of advantages exist for designs such as the one shown in FIG.\",\n \"40, compared to index guiding fiber designs for group velocity dispersion.\",\n \"It is an advantage that the negative slope of 4102 is larger than the negative slope of 4106.This acts to suppress the mode 4104 if the mode-index of the fundamental mode 4103 is near the photonic band gap bottom, when the fundamental mode exhibits large negative group velocity dispersion.\",\n \"It is a further advantage that fibers with a design similar to the one shown in FIG.\",\n \"40 may be designed so that the positive group velocity dispersion of the mode 4104 occurs at wavelengths where the mode-index of 4104 is near the upper photonic band gap edge 4101.This will act to suppress the fundamental mode 4103 so that the positive group velocity dispersion of 4104 can be exploited.\",\n \"It is therefore an advantage that micro-structured fibers with the design shown in FIG.\",\n \"40 may be designed to exhibit large positive group velocity dispersion.\",\n \"This is expected to be of increasing importance as fiber systems with negative group velocity dispersion become more common, in order to e.g.\",\n \"reduce self-phase modulation in fiber systems.\",\n \"The present inventors have realized that it is advantageous to use fibers that guide light by the photonic band gap effect for dispersion compensating purposes.\",\n \"This may seem as a surprise to those skilled in the field, since fibers guiding light by the photonic band gap effect typically have positive dispersion.\",\n \"However, using a dual concentric core arrangement it becomes possible to have a fundamental super mode with negative group velocity dispersion.\",\n \"The present inventors have realized that dual concentric core fibers that guide light by the photonic band gap effect are able to have even larger numerical values of the group velocity dispersion than the group velocity dispersion made possible by the prior art.\",\n \"As indicated by FIG.\",\n \"41 the large values of group velocity dispersion appear near the wavelength where 4105 crosses 4106.Curve 4105 may be seen as the mode-index of the guided mode within the central core 4001 when the outer core 4005 is omitted, and 4106 may correspondingly be seen as the mode-index of the guided mode within the outer core 4005 when the central core 4001 is omitted.\",\n \"Using dual concentric cores for group velocity dispersion the amount of group velocity dispersion that can be obtained is directly linked to the difference between the group velocity of the guided modes with a mode-index corresponding to 4105 and 4106.This difference is maximized when the group velocity corresponding to 4105 is minimized while the group velocity corresponding to 4106 is maximized for a given mode-index, β/k.\",\n \"To investigate the group velocities that can be obtained for guided modes in silica-air fibers, a formula known and understood by those skilled in the art can be applied.\",\n \"v g = c β k ∫ ∏ E × H * · z ⅆ A ∫ ∏ n 2 E × H * · z ⅆ A Here vg is the group velocity, c is the vacuum velocity of light, β k is the mode-index, E is the electric field, H* is the conjugated magnetic field, n is the refractive index of the material at the given place, II is the cross-section where the mode exist, and z is the length direction of the fiber.\",\n \"Assuming as an example that the fiber consist of material with n=1 (air) and n=1.45 (silica) we can rewrite this formula as v g = c β k ∫ air E × H * · z ⅆ A + ∫ silica E × H * · z ⅆ A ∫ air E × H * · z ⅆ A + n silica 2 ∫ silica E × H * · z ⅆ A The formula shows that the group velocity of a guided mode depends on the mode-index, β k , as well as the amount of the modal power that propagates in silica, relative to the modal power that propagates in air.\",\n \"To be more specific the relative amount, fsilica, of the modal power, P = ∫ ∏ E × H * · z ⅆ A , that exist in silica, ∫ silica E × H * · z ⅆ A P , can be used to calculate the group velocity for a guided mode at a given mode-index, such that: v g = c β k 1 + f silica ( n silica 2 - 1 ) From this formula it is seen that a guided mode with the lowest group velocity possible in silica/air will have all its power propagating in silica.\",\n \"Such a mode will have the group velocity v g = c β k n silica 2 .\",\n \"The maximum group velocity possible corresponds to a mode with as much of the field as possible propagating in air.\",\n \"When the mode-index is less than 1 this corresponds to a mode with all its power propagating in air.\",\n \"Such a mode has the group velocity: v g = c β k , where β k ≤ 1.When the mode-index is greater than 1 the maximum group velocity is equal to the phase-velocity of the mode such that: v g = c β / k , where β 2 ≥ 1.The group velocity dispersion, GVD, of a guided mode may be written as: GVD = ⅆ ( 1 v g ) ⅆ λ where λ is the free space wavelength of the mode.\",\n \"The amount of group velocity that can be obtained for a given mode-index can therefore be qualitatively estimated by showing the inverse group velocity, 1 v g as a function of the mode-index, β k .\",\n \"Such a plot is shown in FIG.\",\n \"42, where 4201 shows the inverse group velocity of a mode that propagates solely in silica, 4202 shows a mode that propagates with the maximum group velocity possible.\",\n \"4203 is 4202 subtracted from 4201 and therefore shows the propagation time-difference (in pico-seconds, ps) between the slow 4201 and the fast 4202 mode per kilometre of fiber.\",\n \"Curve 4203 clearly indicates that the propagation time-difference increases strongly as the mode-index, β k is lowered.\",\n \"Since this propagation time-difference is also a measure of the maximum group velocity dispersion, GVD, that can be obtained by using coupled core regions, curve 4203 therefore also indicates that it is advantageous to have a low mode-index, β k , for dispersion compensating purposes.\",\n \"Since this is possible by using photonic band gap technology, curve 4203 therefore shows that it is an advantage to use fibers that guide light by the photonic band gap effect for group velocity dispersion.\",\n \"The present invention therefore also covers photonic crystal fibers with a cladding structure that exhibits photonic band gaps.\",\n \"It is also important to notice that the present invention may be utilized for fiber applications, wherein fibers according to the present invention guides light in higher order modes.\",\n \"The dispersion of higher order mode may be even stronger than the dispersion of the fundamental mode.\",\n \"Apart from the negative dispersion, as a special case, higher order modes in fibers according to the present invention may exhibit very high positive dispersion.\",\n \"Such fibers, guiding light in a higher order mode, and having large positive dispersion may be used for dispersion compensating schemes for fiber-optical communication systems having a negative dispersion over the transmission length.\",\n \"Hence, the present invention also covers dispersion compensating fibers that are guiding light in a higher order mode.\",\n \"Especially, the present invention covers dispersion compensating fibers that are guiding light in a higher order mode with a large positive dispersion.\",\n \"Typically, it is necessary to employ mode converters in systems that utilize dispersion compensating fibers that guide light in a higher order mode.\",\n \"Finally, it should be mentioned, that for both single mode dispersion compensation as well as for higher order mode dispersion compensation, the dispersion compensating fibers are typically packed in modules, and the present invention also covers modules that incorporate fibres being disclosed in the present invention.\",\n \"To fabricate fibers according to the present invention known techniques of stacking and pulling capillary tubes and rods may be used—see for example U.S. Pat.\",\n \"No.\",\n \"5,802,236.FIG.\",\n \"43 shows a schematic example of the cross-section of a preform that may be used to fabricate a fiber with a design as schematically shown in FIG.\",\n \"12a.\",\n \"The preform comprises a central rod 4300 with a high-index material in its center.\",\n \"The rod may have various profiles of the refractive index distribution—for example a step-index profile or a parabolic profile—and it may preferably be fabricated using MCVD techniques.\",\n \"Surrounding the central rod is, for this example, six capillary tubes 4301.The capillary tubes are surrounded by a number of rods including rods 4302 comprising a high-index material.\",\n \"The stack of rods and tubes may be put into a larger over-cladding tube 4304 to yield a final preform.\",\n \"This preform may be drawn into fiber in one or more steps as described for example in U.S. Pat.\",\n \"No.\",\n \"5,802,236.In order to realise different size of voids in the first and the second cladding, it may be advantageous to provide a different pressure to the tubes in the two different regions during one or more steps of the fibre drawing process.\",\n \"In this manner it may for example be advantageous to supply a higher pressure to tubes that define larger voids and a lower pressure to tube that define smaller voids.\",\n \"Depending on the material, of the fiber, drawing temperatures and speeds may vary significantly and experiments may be needed to determine optimum drawing conditions including pressure.\",\n \"Alternatively, the preform may be stacked with tubes of different size, for example different inner diameter.\",\n \"If the preform is drawn at a relatively low temperature, such as around 1850° C.-1900° C. the preform may be drawn to fiber such that the tubes do not collapse.\",\n \"The tubes with smaller inner diameter will thereby result in voids of smaller size.\",\n \"Pressure control of the tubes may be advantageous in providing additional control of the void size.\",\n \"Various manners of applying pressure may be used.\",\n \"It is for example possible to draw the fiber in two steps, wherein a first step tubes are kept open in both ends and the dimensions of the preform is scaled approximately linearly to a cane during drawing in a fiber drawing tower at a temperature of around 1900° C. to 2000° C. The preform may have a diameter of around 1-50 mm and the cane a diameter of around 1-5 mm.\",\n \"In the second step, the cane may be drawn to fiber, where one or more regions are connectorized to a pressure-control chamber such that void size in the final fiber may be controlled.\",\n \"One way of using pressure control for realisation of micro-structured fibers has for example been described by Russell et al.\",\n \"in WO 00/49436.FIG.\",\n \"44 shows two microscope pictures (top and bottom illumination with white light) of a fabricated fiber according to the present invention having a design as schematically shown in FIG.\",\n \"12a.\",\n \"The fibre has Λ of around 4 μm and is designed to exhibit negative dispersion around 1550 nm.\",\n \"The figure also shows the measured dispersion characteristics of the fiber (bottom figure).\",\n \"The fiber is seen to exhibit a dispersion of around −1000 ps/nm/km around 1550 nm—as desirable for dispersion compensating applications.\",\n \"The fiber further has a strong negative dispersion slope with a relative dispersion slope (RDS)—as desirable for dispersion compensation in WDM systems.\",\n \"The measured dispersion characteristic is for a mode confined to the central (or first) core of the fiber.\",\n \"The mode field diameter at 1550 nm is around 4 mm, giving an effective area of around 15 μm2.Hence, the experimental results confirm the simulated results present in the FIGS.\",\n \"16-23.The core comprises a high-index features being separated from the inner cladding features.\",\n \"The core features have a refractive index profile with a maximum refractive index of around 1.47 and a diameter of around 0.7 times the center-to-center spacing of the inner cladding features.\",\n \"The fiber has six high-index features in the outer (or second) core that each has a refractive index profile with a maximum refractive index of around 1.46.The six second core features have a diameter similar to the first core feature.\",\n \"The features in the first and second core are not directly visible in the microscope picture with top illumination (right, top figure) that mainly serves to show the dimensions of the inner cladding features and their position.\",\n \"However, using bottom illumination of the fiber and coupling white light to both the first and second core, the high-index features of the two cores become more visible (left, top figure).\",\n \"It should briefly be mentioned that for operation of the dispersion compensating fibres at near-infrared wavelengths, the mode index of the two cores region is significantly different for light in the visible range (and in according with this and the description of the fibers according to the present invention) and the cores become largely decoupled at the visible where light will be more tightly confined to the high-index features than for the wavelengths of negative dispersion (wavelengths around the previously discussed coupling wavelength regime).\",\n \"FIG.\",\n \"45 shows another example of a real fiber according to the present invention.\",\n \"In comparison with the fiber in FIG.\",\n \"44, the inner cladding features have a smaller size and the dispersion characteristics is seen to exhibit a less negative minimum, but a broader dispersion “dip”.\",\n \"This result is also in agreement with the previously presented simulation results.\",\n \"The fiber presented in FIG.\",\n \"45 has spectrally broad negative dispersion characteristics and a relatively large RDS, hence making the fiber attractive for dispersion compensation of WDM systems operating around 1550 nm.\",\n \"FIG.\",\n \"46 provides a schematic example of a fiber according to the present invention, where the separation between the first core 4600 and the second core 4602 it illustrated.\",\n \"The separation is characterised by the distance d1-2.The first cladding region comprises low-index features 4601 and the second cladding 4602 is homogeneous.\",\n \"The figure further illustrates the refractive index profile along one radial direction in the cross-section of the fiber.\",\n \"The index profile illustrates how the core 4600 is not in contact with the low-index features 4601, but separated by a region 4604.In preferred embodiments, this region comprises a background material being pure silica.\",\n \"The index profile further shows the radial distance from a center of the second core region to a center of the first core region, d1-2.Twice this distance is also referred to as the diameter of the second core.\",\n \"FIG.\",\n \"47 shows a similar illustration as FIG.\",\n \"46, but for a fiber where the second core is formed from an ensemble of high-index features (4700).\",\n \"The use of a separation between the first core and the low-index elements has several advantageous—as for example discussed in connection with FIG.\",\n \"31, and the use of a relatively large diameter of the second core has several advantages as for example discussed in connection with FIG.\",\n \"18.While FIGS.\",\n \"46 and 47 illustrates close to parabolic index profiles of the first and second cores, a broad range of other profiles may be used according to the present invention.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10474030"}}},{"rowIdx":781,"cells":{"text":{"kind":"list like","value":[["Payment system","An electronic transaction payment system is provided having a vendor terminal associated with a vendor who provides goods or services to a purchaser, a vendor smart-card and a vendor smart-card reader for transmitting data to and receiving data from the vendor smart-card.","The system also includes a purchaser smart-card reader which is connected to the vendor terminal and which is operable for transmitting data to and receiving data from a purchaser smart-card.","In operation, payment for goods purchased by the purchaser is made between the purchaser smart-card and the vendor smart-card.","In a preferred embodiment, the payment data is encrypted using an encryption key specific to the transaction between the purchaser and the vendor.","A third party registry terminal may also be provided for providing validation of the vendor and/or the purchaser."],["1.An electronic transaction payment system comprising: a vendor terminal associated with a vendor who provides goods or services to a purchaser; a vendor smart-card; a vendor smart-card reader for transmitting data to and receiving data from said vendor smart-card; a purchaser smart-card; and a purchaser smart-card reader for transmitting data to and receiving data from said purchaser smart-card; wherein said vendor terminal comprises: (i) means for processing requests for vendor goods or services from said purchaser; (ii) means for generating cost data identifying the cost of requested goods or services; (iii) means for interfacing with said vendor smart-card reader; (iv) means for interfacing with said purchaser smart-card reader; and (v) means for transmitting the cost data to said purchaser smart-card via said purchaser smart-card reader interface; wherein said purchaser smart-card comprises: (i) means for receiving said cost data from said vendor terminal; (ii) means for encrypting payment data to be transmitted to said vendor smart-card; and (iii) means 25 for outputting the encrypted payment data for transmission to said vendor smart-card; and wherein said vendor smart-card comprises: (i) means for receiving said encrypted payment data from said purchaser smart-card; and (ii) means for decrypting said encrypted payment data received from said purchaser smart-card to obtain payment for the requested goods or services.","2.A system according to claim 1, wherein said purchaser smart-card further comprises means for digitally signing said payment data using a purchaser digital signature and wherein said vendor smart-card further comprises means for reading the digital signature applied to the payment data to establish the origin of the payment.","3.A system according to claim 2, wherein said vendor smart-card further comprises means for generating receipt data describing the goods or services requested and the payment obtained from said payment data; means for digitally signing said receipt data using a vendor digital signature; and means for outputting the signed receipt data for transmission to said purchaser smart-card; and wherein said purchaser smart-card further comprises means for receiving said signed receipt data and means for reading the vendor digital signature applied to the receipt data to establish the origin of the receipt.","4.A system according to claim 3, wherein at least one of each said digital signatures is generated using a time stamp.","5.A system according to claim 3, wherein at least one of each said digital signatures is generated using a random number.","6.A system according to claim 3, wherein at least one of each said digital signatures is generated using a smart-card identifier.","7.A system according to claim 3, wherein at least one of said digital signatures is generated using a terminal identifier.","8.A system according to claim 3, wherein each of said digital signatures is different.","9.A system according to claim 3, wherein said digital signatures are the same.","10.An electronic transaction payment system according to claim 2, wherein said means for digitally signing said payment data comprises said encrypting means; and said means for reading said purchaser digital signature comprises said decrypting means.","11.A system according to claim 1, wherein: said vendor terminal further comprises means for receiving a request for data describing the vendor; and means for outputting said request to said vendor smart-card; and said vendor smart-card further comprises means for receiving said request for data describing the vendor; means for generating data describing the vendor in accordance with said request; and means for outputting the data describing the vendor to said vendor terminal.","12.A system according to claim 11, wherein said purchaser smart-card further comprises means for receiving said data describing the vendor from said vendor smart-card; and means for determining whether said data describing the vendor conforms to a predetermined condition to determine whether said payment data should be generated for transmission to said vendor smart-card.","13.A system according to claim 1, comprising: a registry; a registry smart-card; and a registry smart-card reader for transmitting data to and receiving data from said registry smart-card; wherein said registry comprises: means for interfacing with said registry smart-card reader; and means for interfacing with said vendor terminal; wherein said purchaser smart-card further comprises means for outputting a request for data describing the vendor for transmission to said registry smart-card; wherein said registry smart-card comprises: means for receiving said request for data describing the vendor from said purchaser smart-card; means for generating data describing the vendor in accordance with said request; and means for outputting the data describing the vendor for transmission to said purchaser smart-card; and said purchaser smart-card further comprises: means for receiving said data describing the vendor from said registry smart-card; and means for determining whether said data describing the vendor conforms to a predetermined condition to determine whether said payment data should be generated for transmission to said vendor smart-card.","14.A system according to claim 1 wherein: said vendor terminal further comprises means for receiving a request for data describing the purchaser; and means for outputting said request for transmission to said purchaser smart-card; and said purchaser smart-card further comprises: means data describing the for receiving said request for data purchaser; for generating data describing the purchaser in accordance with said request; and means for outputting the data describing the purchaser for transmission to said vendor terminal.","15.A system according to claim 14, wherein said vendor smart-card further comprises: means for receiving said data describing the purchaser from said vendor smart-card; and means for determining whether said data 30 describing the purchaser conforms to a predetermined condition to determine whether said goods or services are to be provided to said purchaser or whether said payment data is to be accepted.","16.A system according to claim 1, comprising: a registry; a registry smart-card; and a registry smart-card reader for transmitting data 10 to and receiving data from said registry smart-card; wherein said registry comprises means for interfacing with said registry smart-card reader; and means for interfacing with said vendor terminal; wherein said vendor smart-card further comprises means for outputting a request for data describing the purchaser for transmission to said registry smart-card; wherein said registry smart-card further comprises: means for receiving said request for data describing the purchaser from said vendor smart-card; means for generating data describing the purchaser in accordance with said request; and means for outputting the data describing the purchaser for transmission to said vendor smart-card; and wherein said vendor smart-card further comprises: means for receiving said data describing the purchaser from said registry smart-card; and means for determining whether said data describing the purchaser conforms to the predetermined condition to determine whether said goods or services are to be provided to said purchaser 30 or whether said payment data is to be accepted.","17.A system according to claim 1, wherein said vendor smart-card further comprises a memory for storing an asymmetric encryption key pair comprising a vendor private key and a vendor public key; and means for outputting said vendor public key for transmission to said purchaser smart-card, wherein said purchaser smart-card further comprises means for receiving said vendor public key; and is operable to encrypt said payment data using said vendor public key; and wherein said means for decrypting said payment data in said vendor smart-card is operable to decrypt the payment data using the vendor private key.","18.A system according to claim 17 wherein said asymmetric encryption key pair are generated using an RSA algorithm.","19.A system according to claim 17, wherein said asymmetric encryption key pair are generated using an elliptic curve algorithm.","20.A system according to claim 1, wherein said purchaser smart-card reader interface comprises means for communication via a computer network.","21.A system according to claim 1 wherein said purchaser smart-card reader interface comprises means for communication via the Internet.","22.A system according to claim 1 comprising: a registry; a purchaser registry smart-card associated with the purchaser; and a registry smart-card reader for transmitting data to and receiving data from said purchaser registry smart-card; wherein said registry comprises means for 10 interfacing with said registry smart-card reader; means for interfacing with said vendor terminal; wherein said purchaser smart-card further comprises: means for generating instruction data, for instructing 15 said purchaser registry smart-card to make a payment, to be transmitted to said vendor smart-card; and means for outputting the instruction data for transmission to said purchaser registry smart-card; wherein said purchaser registry smart-card further comprises means for receiving said instruction data from 20 said purchaser smart-card; means for generating payment data to be transmitted to said vendor smart-card; means for encrypting said payment data; and means for outputting the encrypted payment data for transmission to said vendor smart-card; and wherein said vendor smart-25 card further comprises: means for receiving said encrypted payment data from said purchaser registry smart-card.","23.A system according to claim 1, further comprising: a registry; a vendor registry smart-card; and a registry smart-card reader for transmitting data to and receiving data from said vendor registry smart-card; wherein said purchaser smart-card further comprises means for outputting said encrypted payment data for transmission to said vendor registry smart-card; wherein said vendor registry smart-card comprises: means for receiving said encrypted payment data from said purchaser smart-card; means for decrypting said encrypted payment data received from said purchaser smart-card to obtain payment for the requested goods or services; means for generating acknowledgement data describing the obtained payment; and means for outputting said acknowledgement data for transmission to said vendor smart-card; and wherein said vendor smart-card further comprises means for receiving said acknowledgement data from said 20 vendor registry smart-card.","24.A system according to claim 1, further comprising a purchaser terminal associated with the purchaser, the purchaser terminal comprising: (i) means for generating a request for vendor goods or services from said purchaser; (ii) means for interfacing with said purchaser smart-card reader; (iii) means for interfacing with said vendor terminal; and (iv) means for transmitting said purchaser requests to said vendor 30 terminal via said purchaser smart-card.","25.A system according to claim 24, wherein data transmitted between said purchaser terminal and said vendor terminal is encrypted before transmission by the respective smart-cards.","26.An electronic transaction payment system comprising: a vendor terminal associated with a vendor who provides goods or services to a purchaser; a vendor smart-card; a vendor smart-card reader for transmitting data to and receiving data from said vendor smart-card; a purchaser smart-card; and a purchaser smart-card reader for transmitting data to and receiving data from said purchaser smart-card; wherein said vendor terminal comprises: (i) means for processing requests for vendor goods or services from said purchaser; (ii) means for generating cost data identifying the cost of requested goods or services; (iii) means for interfacing with said vendor smart-card reader; (iv) means for interfacing with said purchaser smart-card reader; and (v) means for transmitting the cost data to said purchaser smart-card via said purchaser smart-card interface; wherein said purchaser smart-card comprises: (i) means for receiving said cost data from said vendor terminal; (ii) means for digitally signing payment data to be transmitted to said vendor smart-card; (iii) means for outputting the signed payment data for transmission to said vendor smart-card; wherein said vendor smart-card comprises: (i) means for receiving said signed payment data from said purchaser smart-card to obtain payment for the requested goods or services; (ii) means for reading the digital signature applied to the payment data to establish the origin of the payment; (iii) means for generating receipt data describing the goods or services and the payment obtained from said payment data; (iv) means for digitally signing said receipt data; and (v) means for outputting 10 the signed receipt data for transmission to said purchaser smart-card, wherein said purchaser smart-card further comprises: (iv) means for receiving said signed receipt data; and (v) means for reading the digital signature applied to the receipt data to establish the origin of the receipt.","27.An electronic transaction payment system comprising: a vendor terminal associated with a vendor who provides goods or services to a purchaser; a vendor smart-card; a vendor smart-card reader for transmitting data to and receiving data from said vendor smart-card; a purchaser smart-card; and a purchaser smart-card reader for transmitting data to and receiving data from said purchaser smart-card; wherein said vendor terminal comprises: (i) means for processing requests for vendor goods or services from said purchaser; (ii) means for generating cost data identifying the cost of requested goods or services; (iii) means for interfacing with said vendor smart-card reader; (iv) means for interfacing with said purchaser smart-card; and (v) means for transmitting the cost data to said purchaser smart-card via said purchaser smart-card reader interface; wherein said purchaser smart-card comprises: (i) means for receiving said cost data from said vendor terminal; (ii) means for generating a request for data describing the vendor; and (iii) means for outputting the request for data describing the vendor for transmission 10 to said vendor smart-card; wherein said vendor smart-card comprises: (i) means for receiving said request for data describing the vendor from said purchaser smart-card; (ii) means for generating data describing the vendor in accordance with said request; and (iii) means for outputting the data describing the vendor for transmission to said purchaser smart-card; wherein said purchaser smart-card further comprises: (iv) means for receiving said data describing the vendor from said vendor smart-card; (v) means for determining whether said data describing the vendor conforms to a predetermined condition; (vi) means for, if said data describing the vendor conforms to said predetermined condition, generating payment data to be transmitted to said vendor smart-card; and (vii) means for outputting the payment data for transmission to said vendor smart-card; and wherein said vendor smart-card further comprises: (iv) means for receiving said payment data from said purchaser smart-card to obtain payment for requested goods or services.","28.An electronic transaction payment system comprising: a vendor terminal associated with a vendor who provides goods or services to a purchaser; a vendor smart-card; a vendor smart-card reader for transmitting data to and receiving data from said vendor smart-card; a purchaser smart-card; and a purchaser smart-card reader for transmitting data to and receiving data from said purchaser smart-card; wherein said vendor terminal comprises: (i) means for processing requests for vendor goods or services from said purchaser; (ii) means for generating cost data identifying the cost of requested goods or services; (iii) means for interfacing with said vendor smart-card reader; (iv) means for interfacing with said purchaser smart-card reader; and (v) means for transmitting the 20 cost data to said purchaser smart-card via said purchaser smart-card reader interface; wherein said purchaser smart-card comprises: (i) means for receiving said cost data from said vendor terminal; wherein said vendor smart-card comprises: (i) means for generating a request for data describing the purchaser; and (ii) means for outputting the request for data describing the purchaser for transmission to said purchaser smart-card; wherein said purchaser smart-card further comprises: (ii) means for receiving said request for data describing the purchaser from said vendor smart-card; (iii) means for generating data describing the purchaser in accordance with said request; and (iv) means for outputting the data describing the purchaser for transmission to said vendor smart-card; wherein said vendor smart-card further comprises: (iii) means for receiving said data describing the 10 purchaser from said purchaser smart-card; (iv) means for determining whether said data describing the purchaser conforms to a predetermined condition; (v) means for, if said data describing the purchaser conforms to said 15 predetermined condition, generating acceptance data to be transmitted to said purchaser smart-card; and (vi) means for outputting the acceptance data for transmission to the purchaser smart-card; and wherein said purchaser smart-card further comprises: (v) means for receiving said acceptance data from said vendor smart-card; (vi) means for generating payment data to be transmitted to said vendor smart-card; and (vii) means for outputting the payment data for transmission to said vendor smart-card; and wherein said vendor smart-card further comprises: (viii) means for receiving said payment data from said purchaser smart-card to obtain payment for the requested goods or services.","29.An electronic transaction payment system comprising: a vendor terminal associated with a vendor who provides goods or services to a purchaser; a vendor smart-card; a vendor smart-card reader for transmitting data to and receiving data from said vendor smart-card; a purchaser smart-card; and a purchaser smart-card reader for transmitting data to and receiving data from said purchaser smart-card; wherein said vendor terminal comprises: (i) means for processing requests for vendor goods or services from said purchaser; (ii) means for generating cost data identifying the cost of requested goods or services; (iii) means for interfacing with said vendor smart-card 15 reader; (iv) means for interfacing with said purchaser smart-card; and (v) means for transmitting the cost data to said purchaser smart-card via said purchaser smart-card reader interface; wherein said purchaser smart-card comprises: (i) means for receiving said cost data from said vendor terminal; (ii) means for generating a request for data describing the vendor; and (iii) means for outputting the request for data describing the vendor for transmission to said vendor smart-card; wherein said vendor smart-card comprises: (i) means for receiving said request for data describing the vendor from said purchaser smart-card; (ii) means for receiving data describing the vendor in accordance with said request; and (iii) means for outputting the data describing the vendor for transmission to said purchaser smart-card; wherein said purchaser smart-card further comprises: (iv) means for receiving said data describing the vendor from said vendor smart-card; (v) means for outputting said data describing said vendor to the purchaser; (vi) means for receiving an indication whether or not said data describing the vendor is acceptable to the purchaser; and (vii) means for, if said data describing the vendor is acceptable, causing payment data to be transmitted to said vendor smart-card; and wherein said vendor smart-card further comprises: (iv) means for receiving said payment data to obtain payment for the requested goods or services.","30.An electronic transaction payment system comprising: a vendor terminal associated with a vendor who provides goods or services to a purchaser; a vendor smart-card; a vendor smart-card reader for transmitting data to and receiving data from said vendor smart-card; a purchaser smart-card; and a purchaser smart-card reader for transmitting data to and receiving data from said purchaser smart-card; wherein said vendor terminal comprises: (i) means for processing requests for vendor goods or services from said purchaser; (ii) means for generating cost data identifying the cost of requested goods or services; (iii) means for interfacing with said vendor smart-card reader; (iv) means for interfacing with said purchaser smart-card reader; and (v) means for transmitting the cost data to said purchaser smart-card via said purchaser smart-card reader interface; wherein said purchaser smart-card comprises: (i) means for receiving said cost data from said vendor terminal; wherein said vendor smart-card comprises: (i) means for generating a request for data describing the purchaser; and (ii) means for outputting the request for data describing the purchaser for transmission to said purchaser smart-card; wherein said purchaser smart-card further comprises: (ii) means for receiving said request for data describing the purchaser from said vendor smart-card; (iii) means for receiving data describing the purchaser in accordance with said request; and (iv) means for outputting the data describing the purchaser for transmission to said vendor smart-card; wherein said vendor smart-card further comprises: (iii) means for receiving said data describing the purchaser from said purchaser smart-card; (iv) means for outputting said data describing the purchaser to the vendor; (v) means for receiving an indication whether or not said data describing the purchaser is acceptable to the vendor; (vi) means for, if said data describing the purchaser is acceptable, generating acceptance data to be transmitted to said purchaser smart-card; and (vii) means for outputting the acceptance data for transmission to the purchaser smart-card; and wherein said purchaser smart-card further comprises: (v) means for receiving said acceptance data from said vendor smart-card; (vi) means for generating payment data to be transmitted to said vendor smart-card; and (vii) means for outputting the payment data for transmission to said vendor smart-card; and wherein said vendor smart-card further comprises: (viii) means for receiving said payment data from said purchaser smart-card to obtain payment for the requested goods or services.","31.A smart-card programmed to be able to function as a purchaser smart-card according to claim 1.32.A smart-card programmed to be able to function as a vendor smart-card according to claim 1.33.A smart-card programmed to be able to function as a registry smart-card according to claim 13.34.A smart-card comprises: means for interfacing with a card reader; means for receiving details of a new transaction from an external terminal via said interfacing means and said card reader; means for generating a unique session ID for the new transaction; means for storing the generated session ID; means for receiving message data relating to the new transaction from the external terminal via said card reader and said interfacing means; means for tagging the received message data by appending the stored session ID to the message data; and means for outputting the tagged message data to said external terminal via said interface means and said card reader.","35.A smart-card according to claim 34, further comprising means for encrypting said tagged message data.","36.A smart-card according to claim 34, further comprising means for receiving a session ID generated by another smart-card involved in the new transaction and wherein said tagging means is operable to append the generated session ID and the received session ID to the message data.","37.A smart-card according to claim 34, further comprising a money store for storing electronic money and wherein said tagging means is operable to tag said electronic money with said generated session ID and to output the tagged electronic money back to said external terminal via said interface means in said card reader.","38.A smart-card according to claim 37, comprising means for storing remote account data and wherein said smart-card is operable to receive an instruction identifying whether or not payment for a current transaction is to be made from electronic money stored on the card or from the remote account and wherein said tagging means is operable to tag said electronic money with said session ID if said instruction identifies that the payment should be made from the card and is operable to tag the stored account data if said instruction indicates that the payment is to be made from the remote account.","39.A smart-card according to claim 34, further comprising means for storing user validation data; means for receiving instructions to output user validation data; and wherein said tagging means is operable to tag the user validation data with the session ID prior to outputting the tagged validation data to said external terminal via said interface means and said card reader.","40.A smart-card according to claim 34, further comprising a transaction log store for storing details of data transmitted during each new transaction.","41.A smart-card comprising: means for interfacing with a card reader; a card controller operable for receiving instructions from an external terminal via said interface means and said card reader; means for storing electronic money; means for storing account data relating to a remote account; wherein said card controller is operable to either output electronic money from said money store or to output account details from said account storing means in dependence upon instructions received from an external terminal via said interfacing means and said card reader.","42.A smart-card according to claim 41, wherein said card controller is operable to tag the electronic money or the account data with a unique identifier relating to a current transaction.","43.An electronic transaction system comprising a vendor 15 terminal, a purchaser terminal and a third party registry for providing validation of a purchaser associated with the purchaser terminal and/or validation of a vendor associated with the vendor terminal.","44.A signal carrying processor implementable instructions for causing a smart-card to become programmed as a smart-card according to claim 31.45.A storage medium storing processor implementable instructions for causing a smart-card to become programmed as a smart-card according to claim 31.46.A computer terminal programmed to function as a vendor terminal according to claim 1.47.A computer terminal programmed to function as a purchaser terminal according to claim 24.48.An electronic transaction payment characterized by the step of using an electronic transaction payment system according to claim 1."],["This invention relates to a payment system and in particular but not exclusively to a smart-card payment system wherein an entire purchase operation is explicitly linked as a single transaction and a variable level of user validation is provided.","Existing payment systems suffer from a number of drawbacks.","The use of cash for example is limited by the requirement for the actual transfer of possession of physical tokens and by the fact that if lost or stolen, cash is usable without restriction by a finder or thief.","The use of cheques goes some way to overcome the security worries of carrying cash but still requires the transfer of a physical token.","Cheques also introduce the problem of “clearing time”, that is, time taken by banks to actually transfer money from the cheque writer's account to the cheque receiver's account.","In addition, there is a possibility with a cheque that a bank may refuse to honour a cheque written by an account holder because the account holder has insufficient funds in their account.","In this circumstance, the receiver of the cheque has then transferred ownership of goods or provided a service and has received no payment for those goods or that service.","For ordering goods by telephone or using the internet, cash is totally unsuitable and cheques are very inconvenient because to pay by cheque for goods ordered by telephone or over the internet requires the order placed using the telephone or the internet to be confirmed in writing accompanied by a cheque for payment.","More suitable for use with internet or telephone orders are credit and debit cards.","When using a credit card, payment for the goods is actually made to the retailer by a credit card company which allows the credit card user to spend up to a predetermined borrowing limit for purchases.","Purchases made with a credit card must be paid for to the credit card company within a predetermined time limit or interest on the money borrowed becomes payable.","Debit cards on the other hand provide for an electronic transfer of funds direct from the card holder's bank account to the retailer's bank account.","Credit and debit cards also suffer from the problems of “clearing time”.","Both credit and debit cards use conventional magnetic stripe card technology.","Both payment systems also suffer from processing charges inherent in the system.","These processing charges are typically levied on the vendor and may be in the range of one to four percent of the sale transaction.","Sometimes all or part of this cost is passed onto the purchaser.","This transaction cost has a minimum size which is set by the banks and credit card companies operating the cards.","As a result, it is economically disadvantageous to process small value transactions using credit and debit cards.","Credit and debit cards do however have the advantage for telephone and internet transactions that there is no requirement for the transfer of physical tokens.","They can therefore be used to make purchases or to reserve restaurant tables for example, without the need for postal or personal transfer of payment.","The use of a credit card to make a purchase provides the purchaser with extra protection in that the credit card company must cover the cost of goods paid for on the card, over a certain minimum value, which are not delivered or with which there are certain other problems.","In addition, the credit card operator pays the vendor regardless of whether or not the purchaser ever repays that money to the credit card company.","Thus both vendor and purchaser benefit from the use of a credit card.","However, for telephone and internet transactions, which are defined by the credit card operator as “card holder not present” transactions, the risk of loss rests with the retailer not the card operator.","Thus, it becomes less beneficial for a vendor to accept credit cards as payment for telephone and internet orders because they must pay the handling fee and they receive no protection.","The so-called electronic purse (e.g.","the Mondex Electronic Purse System) has been developed to eliminate the problems of carrying actual cash and of the minimum processing charges for small debit or credit card transactions.","The electronic purse makes use of smart-card or chip-card technology.","To use the electronic purse, the user must either load cash onto the smart-card electronic purse at a vending machine or at the desk of a load agent such as a Post Office, or the user must transfer funds from their bank account or credit card to the smart-card electronic purse.","The purse may then be used to make purchases in the real world, over the internet or over the telephone, subject to the telephone being with an appropriate smart-card reader.","From a transaction processing point of view, full clearing would only be required when loading the purse, which would normally be a medium to large value transaction and therefore acceptable.","Payments could be batched into daily totals and cleared to the retailers as single daily transactions, again being of reasonable size.","There would be no requirement to account for every transaction back to the card holder, all that would happen would be that payment would be made out of the cash float created when taking cash to load the electronic purse.","Thus from a bank's point of view, the electronic purse solved a major problem.","However, from the card holder's point of view the electronic purse is less successful and all open electronic purse trials have tended towards failure in terms of take up.","Among the reasons for the poor take up are: poor education of the card holders, small scale trials leading to purse being usable at only a few locations, lock-in of card holder money to the card and places it could be used, no interest on prepayment, requirement for a bank account to back the electronic purse and fear over security and transaction accountability.","The requirement for a bank account to back the electronic purse is a major failing of the system.","This is because many people who could best benefit from an electronic purse (or other prepaid mechanisms) are those people who do not have bank accounts and who are therefore automatically excluded from using cheques, debit cards or credit cards.","Approximately twenty percent of the UK population are disallowed by the banks from having a bank account.","It should be noted that the Mondex Electronic Purse System was specifically developed to allow use by persons not having a bank account, however the operators still insisted on a user having a bank account before a card would be issued.","Regarding fear over security and transaction accountability, the problem is one of how much the card holder trusts the retailer.","For example, how does the card holder know how much credit is stored on his or her card and how do they know how much is removed?","For internet transactions, this problem grows to incorporate issues such as: how to identify how much is being deducted from the card, how to identify the other party to the transaction, how to identify the location of the other party to the transaction, whether goods will actually be shipped, whether the payment made will actually go to the vendor, whether the vendor will recognise the payment as being against the goods purchased, whether the payment could be fraudulently redirected and concerns over who will be able to see the delivery address specified.","The existing electronic purse schemes use so-called smart-cards or chip-cards.","These are cards having a microprocessor and memory mounted onto or into a card, with an interface provided for the microprocessor to be powered and interfaced with by a reader.","The standards for smart-cards where the interface is by means of physical connection terminals are set out in ISO 7816.The standards for smart-cards where the interface is by means of radio signals (including supply of power) are set out in ISO 14443.In an attempt to address some of the above problems, a number of banks and financial card operators including Mastercard and Visa have agreed a smart-card standard for credit and debit services such that a retailer could have a common terminal to support all smart credit and debit cards in a similar manner to the current situation with magnetic stripe cards.","This standard is known as EMV (Europay Mastercard Visa) and it applies to credit and debit services on smart-cards as well as the terminals to support them.","An electronic purse system based on the EMV specifications is the common electronic purse specifications (CEPS) with which the European Standard Organisation (CEN) has been involved.","CEPS requires a personalised system and additional hardware to identify the cardholder in order for him or her to digitally sign and certify all CEPS transactions.","Further, for use in telephone or internet transactions, there is a requirement in CEPS for digital signatures.","There is currently no worldwide or European standard for producing digital signatures.","Such digital signatures require a public key infrastructure administered by a certification authority which acts as a library or directory of public keys.","In addition, when it supplies a public key it adds a certificate signed with its own private key such that the recipient can then validate that the public key information is genuine within Europe, the various public key infrastructures that currently exist are not compatible and therefore such digital signatures are not interoperable across services and countries.","Thus the full implementation of CEPS will be delayed until such standardisation can be completed.","In summary therefore, cash and cheques require the exchange of physical tokens and are therefore totally unsuitable for real time trading by telephone or over the internet.","Credit and debit cards and cheques suffer from clearing time delays.","Credit and debit cards are also unsuitable for small value transactions due to the processing charges included.","Electronic purses have been unpopular due to the “tied-in” nature of the funds and the small number of places where trials have allowed use of the purses.","Credit and debit cards and electronic purses have the additional problem for internet and telephone transactions of requiring trust between purchaser and vendor.","CEPs has the drawback of an awkward and non-standard security system.","Aspects of the present invention relate to a smart-card payment system wherein: provision is made for payment by means of prepaid value stored on a card held by the purchaser (the so-called electronic purse); provision is made for payment by means of credit by a third party on behalf of the purchaser and accessed by means of a smart-card held by the purchaser (the so-called credit card facility); provision is made for payment by means of prepaid value stored on behalf of the purchaser and accessed by means of a smart-card held by the purchaser (the so-called debit card facility); provision is made for different levels of validation between the purchaser and the vendor including third party validation if required; the provision of a third party registry to permit the movement of value or funds between the registry and the smart-card; payment may be anonymous or personalised; the purchaser may use either of the electronic purse and debit card facilities without having to pass any means test, credit check or hold a bank account; and a system in which an entire purchase operation is inextricably linked as a single transaction and variable levels of user (purchaser and/or vendor) validation is provided.","The present invention seeks to address difficulties of the prior art.","According to one aspect, there is provided an electronic transaction payment system comprising a vendor terminal associated with a vendor who provides goods or services to a purchaser; a vendor smart-card; a vendor smart-card reader for transmitting data to and receiving data from the vendor smart-card; a purchaser smart-card reader for transmitting data to and receiving data from a purchaser smart-card.","The vendor terminal comprises means for processing requests for vendor goods or services from the purchaser, means for generating cost data identifying the cost of requested goods or services and means for transmitting the cost data to the purchaser smart-card.","The purchaser smart-card includes means for receiving the cost data from the vendor terminal and means for encrypting the payment data for transmission back to the vendor smart-card.","The vendor smart-card also includes means for receiving the encrypted payment data and means for decrypting the payment data to obtain payment for the requested goods or services.","Preferably, all communications relating to the purchase of the goods pass from the purchaser's smart-card to the vendor and are encrypted with an encryption key specific to the current transaction.","In this way, payment of the goods and/or services can be inextricably linked to the transaction itself.","The invention can be applied for electronic trading on, for example, the Internet and can be used at traditional points of sale.","In a preferred embodiment, a third party registry is provided to provide guarantees for the purchaser and/or the vendor if required.","The third party registry may provide additional support as requested and to a level requested by the vendor or purchaser, to ensure trust and confidence in the trade.","Either party in the transaction may choose to validate the credentials of the other party by making a reference to the third party registry.","In this case, the registry will provide the information and may levy an appropriate fee.","Preferred embodiments of the present invention will now be described by way of example only with reference to the accompanying drawings, in which: FIG.","1 shows an internet enabled computing environment in which the present invention may operate; FIG.","2 is a block diagram showing the main functional elements of a vendor terminal and a purchaser terminal shown in FIG.","1; FIG.","3 is a block diagram showing the main functional elements of a registry terminal shown in FIG.","1; FIG.","4 is a block diagram showing the main functional elements of a smart-card shown in FIG.","1; FIG.","5 is a flow chart showing the main processing steps performed by the terminals in FIG.","1 to process a transaction; FIG.","6 is a flow diagram showing in more detail the processing steps involved in a product selection and price agreement step which forms part of the processing steps shown in FIG.","5; FIG.","7 which comprises FIGS.","7A and 7B is a flow diagram showing in more detail the processing steps involved in a secure link set-up step which forms part of the processing steps shown in FIG.","5; FIG.","8 is a flow diagram showing in more detail the processing steps involved in a purchaser validation step which forms part of the processing steps shown in FIG.","5; FIG.","9 is a flow diagram showing in more detail the processing steps involved in a vendor validation step which forms part of the processing steps shown in FIG.","5; FIG.","10 is a flow diagram showing in more detail the processing steps involved in a delivery confirmation step which forms part of the processing steps shown in FIG.","5; FIG.","11 which comprises FIGS.","11A and 11B is a flow diagram showing in more detail the processing steps involved in a payment and receipting step which forms part of the processing steps shown in FIG.","5; FIG.","12 which comprises FIGS.","12A and 12B is a flow diagram showing in more detail the processing steps involved in a purchaser validation step which forms part of the processing steps shown in FIG.","5 according to an alternative embodiment; FIG.","13 is a flow diagram showing in more detail the processing steps involved in a purchaser validation step which forms part of the processing steps shown in FIG.","5 according to a further alternative embodiment; FIG.","14 which comprises FIGS.","14A and 14B is a flow diagram showing in more detail the processing steps involved in a vendor validation step which forms part of the processing steps shown in FIG.","5 according to another alternative embodiment; FIG.","15 is a flow diagram showing in more detail the processing steps involved in a payment and receipting step which forms part of the processing steps shown in FIG.","5 according to another alternative embodiment; FIG.","16 is a flow diagram showing in more detail the processing steps involved in a payment and receipting step which forms part of the processing steps shown in FIG.","5 according to another alternative embodiment; FIG.","17 is a flow diagram showing in more detail the processing steps involved in a payment and receipting step which forms part of the processing steps shown in FIG.","5 according to a further alternative embodiment; FIG.","18 which comprises FIGS.","18A and 18B is a flow diagram showing the main processing steps required to convert currency on an account card; FIG.","19 shows an internet enabled face-to-face retail environment in which the present invention may operate; FIG.","20 is a block diagram showing the main functional elements of a retailer terminal shown in FIG.","19.OVERVIEW In the following description it is assumed that the purchaser's terminal is already connected to the internet, as is the vendor's terminal.","Also, using his or her terminal, the purchaser has browsed the vendor's website and has selected a product or products that he or she wishes to purchase.","Referring to FIG.","1, the internet enabled computing environment 1 in which the payment system of the present embodiment is operated, comprises a purchaser terminal 5, a vendor terminal 21 and a registry terminal 25 which communicate electronically with one another via a network such as the internet 19.Attached to the purchaser terminal 5 is a smart-card slot 7p into which a purchaser smart-card 3 may be inserted.","In this embodiment, the purchaser smart-card 3 must be inserted into the smart-card slot 7p in order for any transaction to be made with a vendor.","Also attached to purchaser terminal 5 is a monitor 9 for displaying data to a user, a keyboard 11 and a mouse 13 which provide a user interface to the purchaser terminal 5 and a modem 15 for communicating via the internet 19.An electrical, electromagnetic or optical signal 17 is transmitted from and received by modem 15 in order to facilitate communication via the internet 19.As shown in FIG.","1, the vendor terminal 21 also includes a monitor 9, a keyboard 11, a mouse 13, a modem 15 and a smart-card slot 7v for receiving a vendor smart-card 23.Attached to the registry terminal 25 is a registry smart-card server 27 having a plurality of registry account smart-cards permanently inserted in card slots 7r therein.","In this embodiment, there is typically one registry account logical smart-card for each user of the payment system, although in practice many such logical smart-cards will be grouped onto one master smart-card which will substantially reduce the smart-card and smart-card reader requirement at the registry.","Each registry account is represented by a logical record on a master account card.","The number of master account cards will be equal to the number of accounts divided by the number of logical account records on each card.","Since the information on the card need only contain an account number and balance, with further information held in a connected hard disk at the registry 25, there could be as many as five thousand logical account records per master card.","In this embodiment, every registry account is represented by a disk based record at the registry 25.As and when a registry account is to be accessed for whatever reason, key information is copied to a spare logical account slot in a master card, so that card to card working may take place.","The number of master cards required will be just greater than the number of parallel transactions required to provide adequate performance.","After a transaction is completed, the disk record is updated.","The card record is retained until the logical account slot is required.","This provides a form of caching to provide better performance for those actively using their smart-cards and registry.","Typically, a retailer's account will be more active than a purchaser's account.","FIG.","2 shows the functional elements of the purchaser terminal 5 and the vendor terminal 21.As shown, these terminals include a central processing unit (CPU) 41 which carries out processing operations in accordance with instructions stored in a working memory 43 and in accordance with user input received either via the keyboard and mouse interface 49 or the smart-card reader 53.The terminals also include a monitor interface 51 for interfacing to the display, a modem interface for interfacing with the modem 15 and a non-volatile software store 47 (such as a hard disk) which stores the processing instructions used to control the operation of the CPU 41 together with other data used by the terminals.","As shown, these components are connected together via a bus 45.FIG.","3 shows the functional elements of the registry terminal 25.As shown, the registry terminal 25 also includes a CPU 41, a working memory 43, a software store 47, a monitor interface 51 and a modem interface 55 which are all connected together by a bus 45.In addition, the smart-card server 27 is connected to the registry terminal 25 via a smart-card server interface 57.FIG.","4 shows the functional elements of a smart-card 3,23 used in this embodiment.","As shown, the smart-card 3,23 includes a card controller 62 which carries out processing operations stored in a processing instructions store 64 in accordance with instructions received from the purchaser terminal 5, vendor terminal 21 or the registry terminal 25 via a card reader interface 66.Coupled to the card controller 62 is a money store 68, which holds data representing the amounts of money held on the smart-card 3,23.The smart-card 3,23 also includes a user validation store 70 which stores information about the smart-card owner such as the owner's name, age, address etc.","When a smart-card 3,23 is used to make a transaction, a single unique session ID is generated by a session ID generator 72 on the smart-card 3,23.The generated session ID is encrypted by an encryption engine 74 also on the smart-card 3,23 and then stored in a session ID store 76.As will be described in further detail later, this session ID is appended to all subsequent messages related to the current transaction by the card controller 62, and the card controller 62 is also operable to compare the session ID of received messages to the stored session ID to check that the message belongs to the current transaction.","As shown in FIG.","4, the smart-card 3,23 further comprises a key store 78 for storing data needed for encryption and decryption, such as public and private keys, which will be described in more detail later.","The key store 78 is coupled to the encryption engine 74, so that messages relating to a transaction can be encrypted by the encryption engine 74 using the stored encryption key data, before transmission back to the terminal via the card reader interface.","Also coupled to the card controller 62 is a transaction log store 80 for storing data representing each transaction made using the smart-card 3,23 and a registry data store 82 for storing details of the user account held at the registry.","Referring now to FIG.","5, there are shown the principal functional steps which must be performed to complete a purchase transaction using the payment system of the present embodiment.","As shown, at step S4-1, the user, having selected a desired product already, provides input indicating a product selection to the purchaser terminal 5.The purchaser terminal 5 then communicates with the vendor terminal 21 by transmitting signals 17 between the modems 55 attached to each of the purchaser terminal 5 and the vendor terminal 21 via the internet 19.The purchaser terminal 5 transmits the product selection to the vendor terminal 21, which then returns price and delivery terms to the purchaser 5.The terms are then displayed by the purchaser terminal 5 on monitor 9 and the purchaser terminal 5 then awaits an input from the purchaser to indicate approval or disapproval of those terms via the keyboard 11 or mouse 13.The inputted approval or disapproval is then communicated by the purchaser terminal 5 to the vendor terminal 21 via the internet.","Next, at step S4-3, the purchaser smart-card 3 inserted in the purchaser terminal 5 and vendor smart-card 23 inserted in the vendor terminal 21 create and exchange secure encryption keys (which are stored in the respective key stores 78 of the purchaser and vendor smart-cards 3,23) to be used throughout the payment session and to be used as the basis for the secure exchange of information using data encryption between terminals.","Before payment is made, however, in this embodiment the smart-cards 3,23 return control to the terminals so that user validation can be performed.","This is initiated at step S4-5, where the vendor terminal 21 displays on monitor 9 a request for user input to indicate whether validation of the purchaser should be undertaken.","Such input is received via the keyboard 11 or mouse 13.If validation is to be undertaken, processing proceeds to step S4-7 where validation is performed.","To perform validation, as will be described in more detail below, the vendor terminal 21 communicates with purchaser terminal 5 or the registry 25 via the internet 19 to request identifier data about the purchaser so that certain pre-specified levels of knowledge about the purchaser are obtained.","Following purchaser validation, or if purchaser validation is not to be undertaken, the processing proceeds to step S4-9, where the purchaser's terminal 5 displays on monitor 9 a request for user input to indicate whether validation of the vendor should be undertaken.","If vendor validation is required, then at step S4-11, validation of the vendor is performed, which is a similar process to that described above for purchaser validation.","Following such validation, or if validation of the vendor is not required, processing continues at step S4-13.At this step, it is decided whether or not delivery of the product will be required.","To determine whether delivery is required, the vendor terminal 21 analyses the delivery terms agreed at step S4-1.From these terms it will be clear whether anything is to be delivered.","It is to be understood that delivery may include electronic transfer of data as well as postal delivery of physical items.","If delivery is required, then the delivery data is confirmed at step S4-15 where the purchaser terminal transmits the relevant delivery address (which may be a postal address, an e-mail address or an FTP (file transfer protocol) server address for example) to the vendor terminal 21.Following this, or in the event that delivery is not required, control is passed back to the smart-cards 3,23 where payment and receipting is performed at step S4-17.In this step, the purchaser terminal 5 causes the appropriate value to transfer from the money store 68 of the purchaser smart-card 3 to the money store 68 of the vendor smart-card 23, during which a receipt message is transferred from the vendor smart-card 23 to the purchaser smart-card 3.An overview of the smart-card payment system of this embodiment has been given above.","The system offers a number of advantages, including: i) the provision of a single session ID which links all messages in a transaction including the terms, user validation and payment, thereby inextricably linking the payment to the transaction; ii) the provision of different levels of user (purchaser and/or vendor) validation; and iii) the provision of a third party registry for validation of one or more of the parties to a transaction if required.","These and other advantages of the payment system of the present embodiment will become apparent from the following more detailed description.","Product Selection FIG.","6 shows an expansion of step S4-1 in which the purchaser terminal 5 transfers data describing a product selection to the vendor terminal 23 and the purchaser and vendor terminals 5, 23 communicate to establish the price and delivery terms for the transaction.","In FIG.","6, the steps undertaken by the purchaser's terminal are shown to the left hand side of the central dotted line and those steps undertaken by the vendor's terminal are shown on the right hand side of the central dotted line.","At step S5-1, the purchaser terminal 5 receives product selection input from the user via the keyboard 11 or mouse 13, and at step S5-3 product selection data is transmitted to the vendor terminal 21 by transmitting signals 17 from the modem 55 connected to the purchaser terminal 5 to the modem 55 connected to the vendor terminal 21 via the internet 19.The product selection data is received by the vendor terminal 21 at step S5-5.At step S5-7, the vendor terminal 21 compares the product selection data to a product database to determine the price of the product and at step S5-11 the price and any necessary delivery terms data are transmitted to the purchaser terminal 5.At step S5-13 the purchaser terminal 5 receives the price and delivery terms data, which data is then displayed to a user at step S5-15.The purchaser terminal 5 then receives acceptance input from the user at step S5-17 and at step S5-19 the terminal 5 processes the input from the user to determine whether the user accepted the price and delivery terms.","If the price and delivery terms are not accepted, then at step S5-21 the purchaser terminal 5 transmits that the terms are not accepted to the vendor terminal 21 which is received at step S5-23.The transaction process then ends.","If the user does accept the price and delivery terms, then at step S5-25 the purchaser terminal 5 transmits that the terms have been accepted and at step S5-27 the vendor terminal 21 receives that the terms have been accepted.","Following this, the selection of product and agreement of price and delivery terms is complete and referring again to FIG.","5 processing then continues to step S4-3.Establish Secure Link The processing of step S4-3 is shown in more detail in FIG.","7.At step S6-1, the vendor terminal 21 transmits a request for the purchaser to identify how payment is to be made.","At step S6-3, that request is received at the purchaser terminal 5 and at step S6-5 the purchaser terminal 5 transmits that the payment system of the present embodiment is to be used, ie payment using the smart-cards 3,23.This is received at step S6-7 by the vendor terminal 21.Once it has been established that the payment system of the current embodiment is to be used, control passes to the smart-cards 3,23 which establish a secure session between themselves.","This is achieved, in this embodiment, using so called public key encryption where each party transmits to the other a public key with which data should be encrypted.","It is only possible to decrypt the encrypted data using a private key which is retained by each party.","The present embodiment uses a modified public key encryption scheme which allows a new, unique session ID to be used for each transaction in connection with the asymmetric key pair.","The setting up of the secure session begins at step S6-9 where the vendor smart-card 23 transmits its public key from its key store 78 to the purchaser smart-card 3.The purchaser smart-card 3 receives the public key from the vendor smart-card 23 at step S6-11 and stores it in its key store 78.Next, at step S6-13, the purchaser smart-card 3 generates, using the session ID generator 72, a purchaser session ID which it appends to all communications transmitted to the vendor smart-card 25 during the current session.","In the present embodiment, the purchaser session ID takes the form of a block of data made up of a time stamp, a random number, a terminal ID and a card ID, all encrypted using the purchaser smart-card's public key (so that only the purchaser smart-card 3 can decrypt the purchaser session ID).","Then at step S6-15, the purchaser smart-card 3 encrypts the purchaser session ID using the vendor smart-card's public key and transmits this to the vendor smart-card 23, along with its own public key.","At step S6-17, the vendor smart-card 23 receives and decrypts the data to recover the purchaser session ID and the purchaser public key, which it stores within the session ID store 76 and the key store 78 respectively of the vendor smart-card 23.Next, at step S6-19, the vendor smart-card 23 generates a vendor session ID using its session ID generator 72.In the present embodiment, the vendor session ID comprises a time stamp, a random number, a terminal ID and a card ID, all encrypted using the vendor smart-card's public key (so that only the vendor smart-card 23 can decrypt the vendor session ID).","At step S6-21, the vendor smart-card 23 then encrypts the vendor session ID using the purchaser smart-card's public key and transmits it to the purchaser smart-card 3.At step s6-23, the purchaser smart-card 3 receives and decrypts the data to recover the vendor session ID which it stores in its session ID store 76.At step S6-25, the purchaser smart-card 3 transmits a message to the vendor smart-card 23 stating that the vendor session ID has been received, which message is received by the vendor smart-card 23 at step S6-27.The session ID generated by each of the purchaser smart-card 3 and the vendor smart-card 23 will be different for a given session (because of the time stamp and the random number), but they will be inextricably linked to the one session and the two smart-cards.","In the present embodiment, the session IDs are added to every transfer of data passing between the terminals after it has been established that the payment system of the present embodiment is to be used.","This will take the form of appending both session IDs to the end of the message and then encrypting the whole with the recipient's public key.","By performing the encryption each message is protected from being read by an unauthorised party, and by using the session ID each message is digitally signed to certify that the sender of the data is who they claim to be and that the message is valid.","It should be noted that only the purchaser smart-card 3 and the vendor smart-card 23 will be able to: validate the session IDs, encrypt data and decrypt data.","The cards will be use their own encryption engines 74 for this purpose, rendering it unnecessary for the terminals to be secure devices.","Part of the session will include the transmission of a user input such as acceptance or refusal of validation information supplied and/or delivery data.","This will be entered as plain text (i.e.","unencrypted and uncoded) by the vendor or purchaser and then passed to the local smart-card for appending of session IDs, encryption and transfer to the other party.","Again, it is not necessary for the terminal to be a secure device.","Purchaser Validation Referring now to FIG.","8, the processing steps to perform the validation of a purchaser (step S4-7) will now be described.","First, the vendor or vendor terminal 23 must determine the level of validation required at step S7-1.Note that if it is determined at step S4-5 that no validation is required, then the processing steps shown in FIG.","8 will not be undertaken.","In this embodiment, the level of validation may be selected manually by the vendor, in which case validation options will be displayed by the vendor terminal 21 on monitor 9 and which will then await an input from the vendor via the keyboard 11 or the mouse 13.Alternatively, the level of validation may be selected by the vendor terminal automatically, in which case the characteristics (eg value, nature etc) of the transaction will be compared to a predetermined set of rules for deciding the level of validation required.","Once the level of validation has been determined, the vendor terminal 21 will transmit a request for the purchaser validation at step S7-3 indicating the level of validation required.","In the present embodiment, it is assumed that a relatively low level of validation is required.","Therefore the validation request is sent to the purchaser terminal 5, not the registry 25.At step S7-5, the purchaser terminal 5 receives the validation request and at step S7-7 transmits the requested validation data (which may either be input by the purchaser or is stored in the validation store 70 of the purchaser smart-card 3).","At step S7-9 the vendor terminal 21 receives the validation data and at step S7-11 determines whether the validation data received is acceptable.","This is done either automatically by comparing the received validation data with pre-stored data or manually by displaying the validation data to the vendor and awaiting confirmation from the vendor that it is acceptable.","If the validation data is unacceptable then processing continues at step S7-13 where the vendor terminal 21 transmits to the purchaser terminal 5 that the validation data provided was unacceptable.","This is received at step S7-15 by the purchaser terminal 5 and then the transaction process ends.","Alternatively, if the validation data is acceptable, processing continues at step S7-17 where the vendor terminal 21 transmits to the purchaser terminal 5 that the validation data provided was acceptable and at step S7-19 the purchaser terminal 5 receives this.","The validation of the purchaser is then complete and, referring again to FIG.","5, processing continues at step S4-9.Vendor Validation If it is decided at step S4-9 that validation of the vendor is required, as described above, then the steps shown in FIG.","9 are performed (step S4-11).","The processing of FIG.","9 is substantially the same as the processing of FIG.","8, except that it is the purchaser terminal 5 requesting the validation data and the vendor terminal 21 transmitting it.","Thus at step S8-1, the purchaser terminal 5 determines the level of validation that is required in substantially the same manner as described above with reference to purchaser validation in FIG.","8, following which it transmits a vendor validation request either to the vendor terminal 21 or to the registry 25 at step S8-3.In the present embodiment it is assumed that a relatively low level of validation is required.","Therefore the validation request is sent to the vendor terminal 21, not the registry 25.On receiving the vendor validation request at step S8-5, the vendor terminal 21 transmits, at step S8-7, the requested validation data (which may either be input by the vendor or is stored in the validation store 70 of the vendor smart-card 23).","Following receipt of the validation data by the purchaser terminal 5 at step S8-9, the purchaser terminal 5 determines at step S8-11 whether the validation data is acceptable.","This is done either automatically by comparing the received validation data with pre-stored data or manually by displaying the validation data to the purchaser and awaiting confirmation from the purchaser that it is acceptable.","If the validation data is decided to be unacceptable, then at step S8-13 the purchaser terminal 5 transmits to the vendor terminal 21 that the validation data is not acceptable and the vendor terminal 21 receives this at step SB-15 following which the transaction process terminates.","If, however, the validation data is determined to be acceptable at step S8-11, then at step S8-17 the purchaser terminal 5 transmits to the vendor terminal 21 that the validation data is acceptable.","This acceptance is received by the vendor terminal 21 at step S8-19 following which the validation of the vendor is complete and, referring again to FIG.","5, processing continues at step S4-13.Confirm Delivery Data If it is determined at step S4-13 that delivery is required, then the delivery data must be confirmed at step S4-15.This process is shown in more detail in FIG.","10.At step S9-1, the vendor terminal 21 requests the delivery data, that is the delivery address (which may be a postal address or an e-mail address, for example) and the name of the person or company to whom the items should be directed.","At step S9-3, the purchaser terminal 5 receives the delivery data request and displays on monitor 9, at step S9-5, a request for a user to enter delivery data.","At step S9-7, the purchaser terminal receives the delivery data input from the user via the keyboard 11 or the mouse 13 and at step S9-9 transmits the delivery data to the vendor terminal 21.The vendor terminal 21 then receives the delivery data at step S9-11, following which the process of confirming delivery data is complete and, referring again to FIG.","4, the processing will continue at step S4-17.Payment and Receipting Performing payment and receipting (step S4-17) will now be described in greater detail with reference to FIG.","11.As mentioned above, control of the payment and receipting is carried out in this embodiment by the smart-cards 3,23.Starting at S10-1, the vendor smart-card 23 transmits a request for payment to the purchaser smart-card 3.At step S10-3, the purchaser smart-card 3 receives the request and at step S10-5 transmits the payment data to the vendor smart-card 23.At step S10-7, the vendor smart-card 23 receives the payment data and at step S10-9 transmits a receipt for the payment.","At this time, the vendor smart-card 23 has received the payment data but has not accepted the payment described therein.","The purchaser smart-card 3 then receives the receipt at step S10-11.At step S10-12, the purchaser smart-card 3 validates the receipt by checking that the purchaser session ID has been appended to the receipt data and that it matches its purchaser session ID stored in the session ID store 76.If the purchaser session ID is not present and correct, then at step S10-13, the purchaser smart-card 23 transmits an error message to the vendor smart-card 23 stating that the session ID is incorrect.","This error message is received by the vendor smart-card 23 at step S10-14 following which the transaction process ends.","On the other hand, if it is determined at step S10-12 that the purchaser session ID is present and correct, then at step S10-15 the purchaser smart-card 3 transmits an acknowledgement that the receipt has been received.","This acknowledgement is received by the vendor smart-card 23 at step S10-16, following which the vendor smart-card 23 validates the acknowledgement by checking that the vendor session ID has been appended to the acknowledgement data and that it matches the vendor session ID stored in the session ID store 76.If the vendor session ID is not present and correct, then at step S10-18 the vendor smart-card 23 generates an error message stating that the vendor session ID is not present and transmits it to the purchaser smart-card 3 which receives the message at step S10-19 following which the transaction process ends.","If on the other hand, however, it is determined at step S10-17 that the vendor session ID is present and correct, then processing continues at step S10-20 where the vendor smart-card 23 finalises the order data and accepts the payment data, thereby actually receiving the payment, to complete the transaction.","The receipt and acknowledgement are validated by each smart-card 3 and 23 to ensure the session remains intact.","The transaction can only be completed if both smart-cards 3 and 23 complete this test successfully.","Thus the payment process is inextricably linked to the purchase transaction, the validation of the parties and acceptance of all terms into a single transaction session including user input of validation acceptance and possible delivery details.","Once the session has been validated as complete the vendor smart-card 23 transmits, at steps s10-21, a message to the purchaser terminal 5 to indicate that the transaction is complete.","At step S10-22 the purchaser smart-card 3 receives the transaction complete message following which the transaction process terminates.","Following completion of the transaction, the purchaser terminal 5 and smart-card 3 may then be used to perform other actions unconnected with the purchase.","Validation Level Determination As described above, it is necessary to determine at step S7-1 and S8-1 the level of validation of the other party to a transaction that is required.","For example, one party may wish to validate the address and other credentials (such as age, legal status or credit worthiness) of the other party and also to be guaranteed that purchased goods will be delivered intact and possibly to a given quality.","As discussed above, some of this information, such as name, age or address, is held in the validation store 70 on the smart-card of the user and this information plus further information may be held by the registry 25.Either party may opt for more or less information from the registry 25, and/or a greater or lower level of guarantee.","The registry 25 may implement a charging structure to enable users of the payment system to be charged for information held by and requested from the registry 25, or to charge users for storing and providing their data to others.","Different levels of payment will be required depending on the amount of validation data held by the registry.","In all trades made using the payment system of the present invention, the complete trade, that is selection of goods, agreement to purchase, payment and subsequent expected delivery are based on the honesty of the two parties and their acceptance of each other's bona fides and mutual acceptance of the terms of trading.","The payment system does not apply any specific trading rules of its own and it is up to both parties to agree terms that overcome any doubts a party may have regarding the other party and the trade.","The payment system provides electronic packaging of a negotiation between the parties such that value does not move from one party to the other until both parties are satisfied with the terms.","Thus, each party is able to decide upon the level of trust it has in the other party and if insufficient, to make use of the registry 25 as necessary until satisfied.","For example, two parties trading for the first time may both seek the top level of guarantee from the registry 25.However, as the parties get to know one another, the trust level between them will increase and their reliance on the registry 25 will decrease.","Further Embodiments The first embodiment discussed above made no use of the registry 25 for validation or for payment, as it was assumed that only a low level of validation was required and payment was made directly from the purchaser smart-card 3 to the vender smart-card 23.However, either or both parties to a transaction may wish to validate the other party with the registry 25 or make payment to or from a registry account card 33.Where the registry 25 is to be involved in a transaction, session IDs are generated as described above with reference to FIG.","7, to include the registry (for validation data) and each registry account card used (for payment) in a secure session.","Where the registry 25 is to interact with the purchaser and vendor, a separate secure session with its own session ID pair is generated between the registry 25 and the vendor smart-card 23 and between the registry 25 and the purchaser smart-card 3.However, if only one of the vendor and purchaser needs to contact the registry 25 then a session having a new session ID pair will be generated between the registry 25 and the vendor smart-card 23 or the purchaser smart-card 3.Thus the overall transaction will be made up of a number of sessions, each session having its own session ID pair.","In this embodiment, to ensure a common reference between the various sessions making up the transaction, each party uses the same session ID in each session of a given transaction.","For example, if the purchaser smart-card 3 has already established a session with the vendor smart-card 23 (and has thus created a purchaser session ID), and then requires a session with the registry 25, the purchaser session ID used in the session with the registry 25 will be the same as the purchaser session ID already used in the session with the vendor smart-card 23.Obviously, the vendor session ID and the registry session ID will be different to one another.","Referring now to FIG.","12, there will be described a method of performing validation of the purchaser using validation data held by the registry 25 in which the purchaser is informed of the decision to validate at the registry 25 and has the option to refuse such validation.","Such processing would take the place of the processing described with reference to FIG.","8 above in the first embodiment.","It is assumed in the method shown in FIG.","12 that the registry 25 and the vendor smart-card 23 have already established a secure session by means of a process similar to that set out above with reference to FIG.","7.At step S11-1, the level of validation required is determined as in step S7-1.Next, at step S11-3, the vendor terminal 21 transmits a request for validation at the registry 25 to the purchaser terminal 5.At step S11-5, the purchaser terminal 5 receives the request and at step S11-7 decides whether or not to agree to the level of validation requested.","This decision is either made automatically by the purchaser terminal 5 according to predetermined rules set by the purchaser or manually by presenting the request to the purchaser on monitor 9 and awaiting acceptance or refusal of the request from the purchaser via the keyboard 11 or mouse 9.If the requested level of validation is not acceptable, then processing continues at step S11-9 where the purchaser terminal 5 transmits a disapproval of the validation request to the vendor terminal 21.The disapproval is received by the vendor terminal 21 at step S11-11, following which the transaction process terminates.","On the other hand, if the requested level of validation is agreed, then processing continues at step S11-13, at which step the purchaser terminal 5 transmits an approval of the validation request to the vendor terminal 21.Following receipt of the validation approval at step S11-15, the vendor terminal 21 transmits a validation request (identifying the purchaser) to the registry 25 at step S11-17.At step S11-19, the registry 25 receives the validation request from the vendor terminal 21 and at step S11-21 transmits the appropriate validation data back to the vendor terminal 21.The validation data is received by the vendor terminal 21 at step S11-23.Processing then proceeds as described above with reference to FIG.","8 such that the processing of steps S11-25 to S11-33 corresponds to the processing of steps S7-11 to S7-19.Another alternative to the processing of FIG.","8 is shown in FIG.","13.In this Figure, validation of the purchaser is performed at the registry 25 but the purchaser terminal 5 is given no notice that the vendor terminal 21 is to receive validation data from the registry 25.It is assumed in the following description that the registry 25 and the vendor smart-card 23 have already established a secure session by means of a process similar to the one described with reference to FIG.","7 above.","At step S12-1, the level of validation required is determined as in step S7-1.At step S12-3, the vendor terminal 21 transmits to the registry 25 a request for validation of the purchaser, which request is received by the registry 25 at step S12-5.At step S12-7, the registry 25 transmits the validation data, which is then received by the vendor terminal 21 at step S12-9.Processing then continues as for FIG.","8 with the processing of steps S12-11 to S12-19 corresponding to the processing of steps 7-11 to steps 7-19.A further method of validation will now be described with reference to FIG.","14.In this Figure it is the vendor which is being validated, however it should be understood that all methods of validation can be used to validate either the purchaser or the vendor or both.","It is assumed in the following description that the registry 25 and the vendor smart-card 23 have established a secure session and that the registry 25 and the purchaser smart-card 3 have established a secure session by means of processing similar to that described with reference to FIG.","7 above.","At step S13-1, the level of validation required is determined as for step S8-1.At step S13-3, the purchaser terminal 5 transmits to the registry 25 a request for validation of the vendor.","Following receipt of the validation request by the registry 25 at step S13-5, the registry 25 transmits to the vendor terminal 21 at step S13-7 a request for permission to validate.","The request is received at step S13-9 by the vendor terminal 21, and at step S13-11 it is decided whether to agree to the level of validation requested in substantially the same manner as described above with reference to FIG.","12.If the level of validation is not agreed to then processing continues at step S13-13 where the vendor terminal 21 transmits a disapproval of the validation request to the registry 25, following which the registry 25 receives the validation disapproval at step S13-15.The transaction processing then ends.","On the other hand, if it is decided at step S13-11 that the level of validation requested is agreed, then at step S13-17, the vendor terminal 21 transmits a validation approval to the registry 25, which approval is received by the registry 25 at step S13-19.At step S13-21, the registry 25 then transmits the validation data to the purchaser terminal 5 which receives the validation data at step S13-23.Following this, the processing continues as in FIG.","9 with the processing of steps S13-25 to S13-33 corresponding to the processing of step S8-11 to S8-19.It is also possible for payment to take place in a number of different ways.","For example, as mentioned above, the registry 25 may hold account cards 33p, 33v on behalf of users of the payment system in a smart-card server 27.The presence of these account cards make it possible for a prospective purchaser to have a large volume of funds available to spend using the payment system without having to have those funds tied into the card which they carry, which card could be lost or stolen thereby denying the user access to those funds.","It is therefore possible that a purchaser may wish to make a payment from his or her account card 33p or a vendor may wish to have a payment made to his or her account card 33v.","If such a payment is to take place, then this will be agreed between the parties before payment is made.","If the purchaser or the vendor decides to make payment from or to receive payment from a registry smart-card 33, then the purchaser or the vendor smart-card is informed of this and in response, it provides account details of the appropriate registry smart-card 33, from its registry data store 82.This information is then transmitted to the other smart-card so that the appropriate secure session can be established between the other smart-card and the registry smart-card.","Examples of these will now be described with reference to FIGS.","14 to 16.Referring now to FIG.","15, there will be described payment and receipting steps to replace those described above with reference to FIG.","11 where a purchaser pays from funds held on their smart-card 3 to a registry account smart-card 33v held on the vendors behalf.","It is assumed in the following description that the vendor registry account card 33v and the vendor smart-card 23 have already established a secure session and that the vendor registry account card 33v and the purchaser smart-card 3 have already established a secure session by means of processing similar to that described with reference to FIG.","7 above.","At step S14-1, the vendor smart-card 23 transmits to the purchaser smart-card 3 a request for payment, which request is received at step S14-3.At step S14-5, the purchaser smart-card 3 transmits the appropriate payment to the vendor account card 33v at the vendor account card 33v registry 25, which payment is received by the vendor account card 33v at step S14-7.At step S14-9, the vendor account card 33v then transmits to the vendor smart-card 23 an acknowledgement that the payment has been made which acknowledgement is received at step S14-11 by the vendor smart-card 23.The receipting process now continues as in FIG.","11 with the processing of steps S14-13 to S14-25 corresponding to the processing of steps S10-9 to S10-21.Referring now to FIG.","16, there will be described a method of payment to the vendor's smart-card 23 from the purchaser's registry account card 33p.","It is assumed in the following description that the purchaser smart-card 3 has already transmitted account details of the purchaser's registry account card 33p to the vendor smart-card 23, that the registry account card 33p and the vendor smart-card 23 have established a secure session and that the purchaser registry account card 33p and the purchaser smart-card 3 have already established a secure session by means of processing similar to that described with reference to FIG.","7 above.","At step S15-1, the vendor smart-card 23 requests payment from the purchaser smart-card 3.Following receipt of the request for payment by the purchaser smart-card 3 at step S15-3, the purchaser smart-card 3 transmits, at step S15-5, to the purchaser's registry account card 33p, an instruction to make the payment.","At step S15-7, the registry account card 33p receives the payment instruction and at step S15-9, it transmits the payment to the vendor smart-card 23.The payment is received by the vendor smart-card 23 at step S15-11, following which the receipting procedure proceeds as in FIG.","11, whereby the processing of steps S15-13 to steps S15-25 corresponds to the processing of steps S10-9 to S10-21.Referring now to FIG.","17, there will be described the processing where payment is made from the purchaser's registry account card 33p to the vendor's registry account card 33v.","It is assumed in the following description that the purchaser smart-card 3 and the vendor smart-card 23 have received details of the other's registry account cards 33p,33v; that the two registry account cards 33p and 33v have already established a secure session between each other; that the purchaser registry account card 33p and the purchaser smart-card 3 have already established a secure session between each other; and that the vendor registry account card 33v and the vendor smart-card 23 have already established a secure session between each other.","At step S16-1, the vendor smart-card 23 transmits a request for payment to the purchaser smart-card 3, which request is received at step S16-3.At step S16-5, the purchaser smart-card 3 transmits a payment instruction to the purchaser's registry account card 33p, which payment instruction is received at step S16-7.The purchaser account card 33p then effects the payment to vendor's registry account card 33v at step S16-9.At step S16-11, the vendor's registry account card 33v transmits a payment acknowledgement to the vendor smart-card 23 which acknowledgement is received at step S16-13.Thereafter, the receipting process between the purchaser smart-card 3 and the vendor smart-card 23 continues as in FIG.","11, with the processing of steps S16-15 to S16-27 corresponding to the processing of steps S10-9 to S10-21.A further optional feature of the payment system described in any of the above embodiments is that a smart-card for use with the payment system may hold funds in a number of different currencies.","It will be possible to load funds onto a purchaser's smart-card 3 or registry account card 33p in the required currency or it would be possible to convert funds held on the card.","Conversion between currencies could take place on a particular user card 3 or 23 or, as would be preferable from a fraud control point of view, conversion could be undertaken on a user's registry account card 33 under the control of the registry 25 to prevent fraudulent currency creation.","The process of currency conversation does not entail the actual conversion of one currency into another, rather it implies debiting an amount in one currency and crediting the equivalent amount in another currency.","The implications of this are twofold; 1) external (off card) funds must be held in the currency to be supplied; 2) the exchange rate will be externally (off card) supplied.","An example of how conversion of funds within a registry account card 33 could be accomplished by a user will now be described with reference to FIG.","18.Before the currency conversion process may commence, a secure session between the user's own card 3 or 23 and the registry card 33p or 33v, respectively must be established.","This is achieved in the same manner as described above with reference to FIG.","6.Thus steps S17-1 to S17-19 correspond to steps S6-9 to S6-27 in that the user card 3 transmits its public key to the registry card which, following receipt of the user card public key, generates a registry session ID in the same manner as the session IDs were generated with reference to FIG.","7 above.","The registry smart-card 33p or 33v then encrypts the registry session ID with the cardholder public key and transmits the encrypted session ID and the registry public key back to the user smart-card 3.The user smart-card 3 then generates a user session ID, encrypts it with the registry public key and transmits the encrypted user session ID to the registry.","Once the registry smart-card 33p or 33v has received the encrypted user session ID, it transmits a message to the user card 3 stating that the session ID was received successfully.","Following this, at step S17-21 the cardholder terminal 5 transmits a request for conversion of account funds from a first currency to a second currency which request is received by the registry 25 at step S17-23.At step S17-25, the registry 25 performs a check to determine whether the account card 33 holds sufficient funds to perform the currency conversion.","If it is determined that sufficient funds are not present, then processing continues at step S17-27, where the registry 25 transmits to the cardholder terminal 5 that there are insufficient funds in the account card 33p to perform a conversion.","This is received by the cardholder terminal 5 at step S17-29, following which the conversion procedure terminates.","On the other hand, if it is determined at step S17-25 that there are sufficient funds in the account to perform the currency conversion then at step S17-31 the funds are converted and at step S17-33 the registry 25 transmits to the cardholder terminal 5 a confirmation that the funds have been converted and a note of the funds totals in the relevant currencies available on the registry account card following the conversion.","This is received by the card holder terminal 5 at step S17-35, following which the currency conversion procedure terminates.","A further feature which may be applied to the payment system according to any of the described embodiments is that transaction logging may be undertaken.","At the end of a given transaction, both the vendor and purchaser will have stored in the transaction log store 80 of their respective smart-cards 3, 23 a record of all of the data transferred during the transaction.","This record will include the goods or services specified, any validation data concerning the purchaser or vendor, any delivery data and payment data.","Each party to the transaction therefore has a complete record of that transaction.","A limited number of such transaction records may be held by the smart-card 3, 23 to provide a short-term transaction log.","However, due to the memory limitations of smart-cards, it is not likely to be possible for the smart-card to store a complete log of all transactions in which the particular smart-card 3, 23 is involved.","Therefore, the smart-cards are programmed to allow the transaction records to be copied from the transaction log store 80 into a full transaction log within a personal computer or the like.","For example, a vendor may set up an automatic process through their usual vending terminal such that at the completion of any transaction, a copy of that transaction record is made on a central transaction log held within the vendor terminal or in a further terminal which is connected to the vendor terminal.","A purchaser may for example, have a home computer comprising a smart-card reader into which the user smart-card 3 may be inserted such that records of purchases made using the smart-card 3 may be transferred to a log on the home computer.","In addition, the registry 25 may be configured to automatically save details of all transactions in which it is involved in order to provide a centralised accountability for the payment system.","In the above embodiments, different levels of validation were described.","Some practical trading examples will now be described that illustrate how these different levels of validation would be used.","VALIDATION LEVEL EXAMPLES Internet Trading Example 1 In this example, a user of the payment system wishes to purchase weather forecast data from the UK Meteorological Office.","The purchaser knows he is at the Meteorological Office website and trusts them to deliver the requested information.","There is no reason for the purchaser to carry out any further checking on the vendor.","The Meteorological Office does not care who the purchaser is or where they are located.","The information to be purchased is not restricted in any way.","In addition, the Meteorological Office will collect payment from the purchasers smart-card before shipping the requested information.","So the vendor requires no further information about the purchaser and an immediate anonymous trade can take place subject to both parties indicating their satisfaction with the price and terms of trade.","Internet Trading Example 2 In this example, the purchaser wishes to buy videos from an internet discount store.","In this case the value of the trade will be low but checking is required by both parties.","The vendor will wish to assure itself that the purchaser is of a suitable age to purchase the videos requested, while the purchaser will wish to validate the credentials and location of the vendor to ensure that the vendor will not simply take the purchaser's money and disappear, never delivering the goods.","The vendor could simply ask a purchaser for their age and address but there would be no support for this information, which therefore could not be trusted.","The vendor would therefore check the credentials of the purchaser with the registry 25 and if the purchaser was not registered or refused to permit the information to be released, the vendor could refuse to sell the products to this purchaser.","If the information were provided, only a low confidence level would probably be required and the registry 25 would provide no warranties or guarantees.","The purchaser would wish to be assured that the vendor can be trusted.","To achieve this, the purchaser would check that the vendor had a registry account and that the vendor would allow the registry 25 to release/confirm the address of the vendor.","Only under these circumstances would the purchaser agree to make the trade.","When both parties are satisfied with the information supplied, the sale price, the transaction charges and the terms of trading, the sale would be completed and value transferred.","In subsequent trades between the two parties, it is likely that the purchaser will have built up a level of trust in the vendor such that the purchaser will not seek any validation from the registry.","Internet Trading Example 3 This example concerns the purchase of fine art.","In this circumstance a great deal of checking and validation will be required.","Even though the vendor will be paid immediately, the vendor may need to verify the nationality and domicile of a purchaser.","The vendor may also wish to avoid being part of a possible money laundering exercise and will therefore seek the most exhaustive credentials concerning a purchaser with the highest level of confidence.","The purchaser will also need to have more guarantees than simply the credentials of the vendor.","Since the fine art cannot be authenticated until it is shipped (or the benefit of using the internet will be lost), and since the goods may be damaged in transit, the purchaser may seek a cash back guarantee from the smart-card registry 25.For this purpose, the registry 25 may be underwritten to supply such a guarantee with the properly authenticated vendors but of course is likely to levy a high transaction charge for this service.","Once all parties have agreed to the levels of confidence they have been given, agreed prices, transaction charges and terms and conditions, the trade may be completed.","Given the high value of the deal, it is likely that payment will be made from an account card held by the registry on the purchaser's behalf, rather than his or her personal card.","In addition, it may be possible for the registry operator to function as a credit providing service such that users of the payment system may be able to borrow money from the registry to be paid back at a later date as with a traditional credit card 25.Real World Trading Example 1 For the purpose of purchasing goods such as a newspaper the smart-card of the payment system according to the present invention may be used as a “traditional” electronic purse at any shop accepting the card.","To make a newspaper purchase the cardholder will simply use his or her card in an anonymous manner to make the purchase using a purchaser smart-card reader provided within the shop.","Real World Trading Example 2 For the purchase of a larger value item such as a television, the purchaser may have sufficient value on their personal card to make payment or they may indicate their desire to pay from the registry account card in much the same way as a traditional bank debit card payment.","In either event, the vendor will wish to validate the credentials of the purchaser to check that the card has not been lost or stolen and that a registry account is not being fraudulently used.","In the case of account payment, the vendor will also take payment from the registry 25.For this purpose the vendor will need a simple smart-card terminal capable of connectivity either directly to the registry or via the internet.","Such terminal may be similar to the bank/credit card terminal currently in use for debit and credit card payments.","Since the purchaser is physically in the vendor's shop it is unlikely that they will seek any further accreditation of the vendor, however this is always possible.","For example, the purchaser may wish to obtain guarantees about the delivery date and goods quality for purpose made goods.","To achieve this, the purchaser will make use of a vendor's terminal to contact the registry and seek the necessary guarantees.","Once all terms have been agreed, the purchaser can authorise payment to be made in exactly the same way as in the case with internet trading.","Real World Trading Example 3 Booking or purchasing a ticket from an unattended terminal is the same as internet trading.","For ticket purchases, it will be possible for a physical paper ticket to be issued via the terminal, for a ticket to be delivered to the vendor following the purchase transaction at the terminal or for an electronic ticket to be written to the purchaser's smart-card.","This is possible because a smart-card is not restricted to holding programs and/or data for a single purpose but may have a number of separate application entities on the same card.","In particular, technologies such as Sun Microsystems Inc's JavaCard™ technology allow applications to be written to a smart-card at any time.","Thus the user of a smart-card equipped with programming to perform processing to enable it to participate in the payment system of the present invention could decide that the ability to have electronic tickets written to his or her card would be useful and therefore arrange for software to enable that function to be written to his or her card at any time.","Although it has been described above with reference to FIGS.","1 to 16 that the purchaser terminal 5 and the vendor terminal 21 are physically separated and connected via the internet, it is possible that a cardholder may wish to use their smart-card at the vendor's location.","This situation is discussed in the above trading examples, and is shown in a simplified form in FIG.","19.In FIG.","19, the vendor terminal 21 having a monitor 9, a keyboard 11 and a mouse 13, which terminal may be combined with or a part of a shop cash register or till, is equipped with a smart-card reader 31 into which the vendor smart-card 23 may be inserted.","For security purposes it may be preferable for the smart-card slot 31 to be physically separated from the terminal 21 and connected via a cable or other data communications link because the terminal 21 may be in an area accessible to customers and having the smart-card 23 in such a location could be perceived to be a security risk.","As before, the vendor terminal 21 is equipped with a modem 15 by means of which it may communicate with the registry terminal 25 by transmitting signals 17 via the internet 19 or by a telephone line.","Also connected to the vendor terminal 21 is a user smart-card reader 34 into which the user smart-card 3 may be inserted by a purchaser when a purchase is to be made using the payment system of the present embodiment.","In such circumstances, there is obviously no need for communication between the purchaser smart-card 3 and the vendor smart-card 23 to take place via the internet and thus the same process as described above with reference to flow diagrams 4 to 16 may be performed as shown but with communication between the cards 3 and 23 taking place via the cables connecting the user smart-card reader 34 to the vendor smart-card reader 31.It should be noted that in the above-described arrangement, the vendor and purchaser smart-cards still create a secure session to wrap the transaction to provide full transaction accountability and traceability, to provide a secure channel between the cards to transfer funds, and to enable data to be certified, all as described above.","In the event that either the purchaser or the vendor should wish to validate the other party at the registry or make payment from or receive payment to a registry account card this may be achieved using the internet connection to the registry terminal 25.Referring to FIG.","20, there is shown a block diagram of the main functional components of the vendor terminal 21 used in this embodiment.","As before, a CPU making use of a working memory 43 to perform instructions stored in software store 47, a keyboard and mouse interface 49 to receive input from a user and a monitor interface 51 to output information to a user are present and are connected via a data bus 45.Also connected via the data bus 45 is the modem 55 for communication with the registry terminal 25 via the internet 19 or telephone line.","The vendor smart-card reader interface 59 and the purchaser smart-card reader 61 are also connected to the data bus 45.Although it has been described above that the various terminals communicate with one another via the internet, the terminals may be connected together via a Local Area Network (LAN) or a Wide Area Network (WAN).","In addition, it is possible that the terminals could perform a so-called “tunnelling” operation to create a Virtual Private Network (VPN) between the terminals across the internet, or a direct dial telephone link between the terminals may be used.","Although it has been described above that where particular validation data or particular delivery terms are decided to be unacceptable the transaction process terminates, further negotiations between the two parties could be undertaken to resolve such situations.","For example, a prospective purchaser may select a product and request next day delivery, but then decide that the cost for next day delivery is more than he or she is willing to pay.","It could be made possible for the prospective purchaser to request to be offered a cheaper delivery service which they might then accept.","Although it has been described above, particularly with reference to FIG.","5, that validation of a purchaser is performed before validation of the vendor, it is not essential that the validations are performed in this order and therefore validation of a vendor could be performed before validation of the purchaser or even at the same time.","Although, it has been described above with reference to FIG.","5 that a step is taken to determine whether or not to validate the purchaser or vendor followed by which the level of validation required is determined, it is possible that these two steps could be combined into one single operation, wherein if no validation is required the steps to achieve validation could be omitted.","Although it is described above with reference to FIG.","9 that a purchaser is required to enter delivery data when required, it is possible that such data could be stored within the purchaser terminal or purchaser smart-card and the transfer of such data to the purchaser terminal could be achieved without reference to the purchaser.","Although it is described above that the registry 25 stores validation data, it is possible that the validation data could be stored on the registry account card 33v, 33p.","Although it is described above that a single registry 25 is present, it is possible that a number of registries could exist.","In this case it would become necessary to establish which registry should be queried to receive validation data from the party to be validated.","For simplicity, this information is preferably stored on the party's smart-card.","Although it has been described with reference to FIG.","4 that delivery data is transmitted after validation, it is entirely possible to include delivery data within the validation data.","This is a favourable modification to the above system as it is likely that validation data would include address data and therefore duplication of data can be reduced and thus transaction processing time can be reduced.","The delivery data would always be transmitted after establishing the session, but need not necessarily be encrypted.","Although it has been described above with reference to FIG.","7 that a specific security arrangement is used to establish the secure session, it is to be noted that the security algorithms used by the smart-cards and the operation of the security engine on the smart-cards does not matter, provided that an asymmetric key system of the type used in public key encryption is used.","Examples of encryption schemes which could be used are RSA type algorithms and elliptic curve techniques.","However, it is not an open public key system requiring a certification authority to validate ownership, rather it is a private system known only to the smart-cards themselves.","Although it has been described above with reference to FIG.","7 that all data passing between the terminals (and the registry) will be encrypted by the smart-card 3, 23, 33 in the terminal (or registry) before transmission, it is not necessary for all data to be encrypted.","Only the data which the parties do not wish to be general public knowledge (which could happen if the data were intercepted) and the data which requires certification as to its origin and validity (such as validation data), and therefore requires a “digital signature”, need be encrypted.","As those skilled in the art will appreciate, the function of the secure session IDs of digitally signing the transmitted data to certify validity and origin could be achieved by methods other than that described above.","In particular, a number of techniques of so-called “digital water-marking” exist, which could be used to certify the data, or encryption keys themselves could be used as signatures.","Although it has been described above that the various terminals have specific components and arrangements, it is sufficient for the performance of the embodiments that any stand alone terminal has a card reader, a means for presenting data to and requesting data from a user, means for receiving user input and means for communicating with other terminals.","Thus it is not necessary for the terminal to be a desk-top computer.","It is possible that terminals may take the form of railway, airline or bus ticket vending machines, public information booths or terminals, theatre or cinema ticketing booths, shop tills, portable computers, hand-held computers and information appliances, telephones and tourist attraction entry gates, for example.","As those skilled in the art will appreciate, the present invention can be used not only from a purchaser's own home terminal but from terminals in so-called internet cafes and in other public terminals.","Although it has been described above with reference to FIGS.","11 to 16 that the registry or a registry account card is included in the secure session before a request for validation data or payment is made to the registry or account card, it is possible that the registry or registry account card will only be included in the secure session when validation data or payment is requested.","Although it has been described above that a user would hold funds on their user smart-card and may hold further funds on their registry account card, it is possible to operate the entire system in the manner of a credit card system whereby a user holds no funds on their own smart-card or on their registry account card, but every time they wish to make a purchase requesting payment from the registry account, and funds being provided to the registry account by the registry on a credit basis with the money supplied as credit to be repaid by the user according to predetermined practices.","It is also possible for the system to be operated in a combination of a stored cash and credit based system whereby a user may store small amounts of cash on their personal smart-card or on their registry account card, but for large purchases make use of a credit arrangement with the registry provider.","Such a system would address the problem of previous smart-card systems whereby funds held on the card are “locked in” to the card.","The smart-cards used to implement the present invention may use any standard interface.","There is an International Standard (ISO7816-3) which governs the so-called contact interface where the microchip on the smart-card interfaces with a reader by means of physical contacts between the microchip on the smart-card and the reader.","There is a further International Standard (ISO14443) which governs the so-called contactless interface.","In this interface, the microchip on the smart-card and the reader have no physical connection, all communication between the reader and the smart-card, including supply of power to the smart-card, is performed using radio transmissions.","It is also possible to implement the invention using a smart-card capable of using both of the above interface types.","As described above, the smart-cards 3,23,33 may be implemented using multi-programmable technologies such as Sun Microsystems Inc's JavaCard™ technology.","It is therefore possible for a person or organisation already possessing a smart-card conforming to such a multi-programmable technology to have software to enable the smart-card as a smart-card 3,23,33 according to any of the above embodiments loaded onto their smart-card.","Such software loading could be performed by an authority such as a registry operator, or the software could be transmitted to the smart-card owner via the internet or similar connection, or on a storage medium such as a floppy disk or CD-ROM for the smart-card owner to use to load the software onto the smart-card themselves.","Although it has been described above that the session IDs should be generated using a time stamp, a random number, a terminal ID and a smart-card ID, it is not necessary that the session ID be generated in this manner for the successful operation of the embodiments.","As one skilled in the art would appreciate, any method of generating a new, unique session ID for each transaction that a particular smart-card 3,23,33 is involved in, could be used.","This method could involve using less than all of the above parts of the session ID described in the above embodiment, or other values such as account holder identifiers or a transaction counter could be used."]],"string":"[\n [\n \"Payment system\",\n \"An electronic transaction payment system is provided having a vendor terminal associated with a vendor who provides goods or services to a purchaser, a vendor smart-card and a vendor smart-card reader for transmitting data to and receiving data from the vendor smart-card.\",\n \"The system also includes a purchaser smart-card reader which is connected to the vendor terminal and which is operable for transmitting data to and receiving data from a purchaser smart-card.\",\n \"In operation, payment for goods purchased by the purchaser is made between the purchaser smart-card and the vendor smart-card.\",\n \"In a preferred embodiment, the payment data is encrypted using an encryption key specific to the transaction between the purchaser and the vendor.\",\n \"A third party registry terminal may also be provided for providing validation of the vendor and/or the purchaser.\"\n ],\n [\n \"1.An electronic transaction payment system comprising: a vendor terminal associated with a vendor who provides goods or services to a purchaser; a vendor smart-card; a vendor smart-card reader for transmitting data to and receiving data from said vendor smart-card; a purchaser smart-card; and a purchaser smart-card reader for transmitting data to and receiving data from said purchaser smart-card; wherein said vendor terminal comprises: (i) means for processing requests for vendor goods or services from said purchaser; (ii) means for generating cost data identifying the cost of requested goods or services; (iii) means for interfacing with said vendor smart-card reader; (iv) means for interfacing with said purchaser smart-card reader; and (v) means for transmitting the cost data to said purchaser smart-card via said purchaser smart-card reader interface; wherein said purchaser smart-card comprises: (i) means for receiving said cost data from said vendor terminal; (ii) means for encrypting payment data to be transmitted to said vendor smart-card; and (iii) means 25 for outputting the encrypted payment data for transmission to said vendor smart-card; and wherein said vendor smart-card comprises: (i) means for receiving said encrypted payment data from said purchaser smart-card; and (ii) means for decrypting said encrypted payment data received from said purchaser smart-card to obtain payment for the requested goods or services.\",\n \"2.A system according to claim 1, wherein said purchaser smart-card further comprises means for digitally signing said payment data using a purchaser digital signature and wherein said vendor smart-card further comprises means for reading the digital signature applied to the payment data to establish the origin of the payment.\",\n \"3.A system according to claim 2, wherein said vendor smart-card further comprises means for generating receipt data describing the goods or services requested and the payment obtained from said payment data; means for digitally signing said receipt data using a vendor digital signature; and means for outputting the signed receipt data for transmission to said purchaser smart-card; and wherein said purchaser smart-card further comprises means for receiving said signed receipt data and means for reading the vendor digital signature applied to the receipt data to establish the origin of the receipt.\",\n \"4.A system according to claim 3, wherein at least one of each said digital signatures is generated using a time stamp.\",\n \"5.A system according to claim 3, wherein at least one of each said digital signatures is generated using a random number.\",\n \"6.A system according to claim 3, wherein at least one of each said digital signatures is generated using a smart-card identifier.\",\n \"7.A system according to claim 3, wherein at least one of said digital signatures is generated using a terminal identifier.\",\n \"8.A system according to claim 3, wherein each of said digital signatures is different.\",\n \"9.A system according to claim 3, wherein said digital signatures are the same.\",\n \"10.An electronic transaction payment system according to claim 2, wherein said means for digitally signing said payment data comprises said encrypting means; and said means for reading said purchaser digital signature comprises said decrypting means.\",\n \"11.A system according to claim 1, wherein: said vendor terminal further comprises means for receiving a request for data describing the vendor; and means for outputting said request to said vendor smart-card; and said vendor smart-card further comprises means for receiving said request for data describing the vendor; means for generating data describing the vendor in accordance with said request; and means for outputting the data describing the vendor to said vendor terminal.\",\n \"12.A system according to claim 11, wherein said purchaser smart-card further comprises means for receiving said data describing the vendor from said vendor smart-card; and means for determining whether said data describing the vendor conforms to a predetermined condition to determine whether said payment data should be generated for transmission to said vendor smart-card.\",\n \"13.A system according to claim 1, comprising: a registry; a registry smart-card; and a registry smart-card reader for transmitting data to and receiving data from said registry smart-card; wherein said registry comprises: means for interfacing with said registry smart-card reader; and means for interfacing with said vendor terminal; wherein said purchaser smart-card further comprises means for outputting a request for data describing the vendor for transmission to said registry smart-card; wherein said registry smart-card comprises: means for receiving said request for data describing the vendor from said purchaser smart-card; means for generating data describing the vendor in accordance with said request; and means for outputting the data describing the vendor for transmission to said purchaser smart-card; and said purchaser smart-card further comprises: means for receiving said data describing the vendor from said registry smart-card; and means for determining whether said data describing the vendor conforms to a predetermined condition to determine whether said payment data should be generated for transmission to said vendor smart-card.\",\n \"14.A system according to claim 1 wherein: said vendor terminal further comprises means for receiving a request for data describing the purchaser; and means for outputting said request for transmission to said purchaser smart-card; and said purchaser smart-card further comprises: means data describing the for receiving said request for data purchaser; for generating data describing the purchaser in accordance with said request; and means for outputting the data describing the purchaser for transmission to said vendor terminal.\",\n \"15.A system according to claim 14, wherein said vendor smart-card further comprises: means for receiving said data describing the purchaser from said vendor smart-card; and means for determining whether said data 30 describing the purchaser conforms to a predetermined condition to determine whether said goods or services are to be provided to said purchaser or whether said payment data is to be accepted.\",\n \"16.A system according to claim 1, comprising: a registry; a registry smart-card; and a registry smart-card reader for transmitting data 10 to and receiving data from said registry smart-card; wherein said registry comprises means for interfacing with said registry smart-card reader; and means for interfacing with said vendor terminal; wherein said vendor smart-card further comprises means for outputting a request for data describing the purchaser for transmission to said registry smart-card; wherein said registry smart-card further comprises: means for receiving said request for data describing the purchaser from said vendor smart-card; means for generating data describing the purchaser in accordance with said request; and means for outputting the data describing the purchaser for transmission to said vendor smart-card; and wherein said vendor smart-card further comprises: means for receiving said data describing the purchaser from said registry smart-card; and means for determining whether said data describing the purchaser conforms to the predetermined condition to determine whether said goods or services are to be provided to said purchaser 30 or whether said payment data is to be accepted.\",\n \"17.A system according to claim 1, wherein said vendor smart-card further comprises a memory for storing an asymmetric encryption key pair comprising a vendor private key and a vendor public key; and means for outputting said vendor public key for transmission to said purchaser smart-card, wherein said purchaser smart-card further comprises means for receiving said vendor public key; and is operable to encrypt said payment data using said vendor public key; and wherein said means for decrypting said payment data in said vendor smart-card is operable to decrypt the payment data using the vendor private key.\",\n \"18.A system according to claim 17 wherein said asymmetric encryption key pair are generated using an RSA algorithm.\",\n \"19.A system according to claim 17, wherein said asymmetric encryption key pair are generated using an elliptic curve algorithm.\",\n \"20.A system according to claim 1, wherein said purchaser smart-card reader interface comprises means for communication via a computer network.\",\n \"21.A system according to claim 1 wherein said purchaser smart-card reader interface comprises means for communication via the Internet.\",\n \"22.A system according to claim 1 comprising: a registry; a purchaser registry smart-card associated with the purchaser; and a registry smart-card reader for transmitting data to and receiving data from said purchaser registry smart-card; wherein said registry comprises means for 10 interfacing with said registry smart-card reader; means for interfacing with said vendor terminal; wherein said purchaser smart-card further comprises: means for generating instruction data, for instructing 15 said purchaser registry smart-card to make a payment, to be transmitted to said vendor smart-card; and means for outputting the instruction data for transmission to said purchaser registry smart-card; wherein said purchaser registry smart-card further comprises means for receiving said instruction data from 20 said purchaser smart-card; means for generating payment data to be transmitted to said vendor smart-card; means for encrypting said payment data; and means for outputting the encrypted payment data for transmission to said vendor smart-card; and wherein said vendor smart-25 card further comprises: means for receiving said encrypted payment data from said purchaser registry smart-card.\",\n \"23.A system according to claim 1, further comprising: a registry; a vendor registry smart-card; and a registry smart-card reader for transmitting data to and receiving data from said vendor registry smart-card; wherein said purchaser smart-card further comprises means for outputting said encrypted payment data for transmission to said vendor registry smart-card; wherein said vendor registry smart-card comprises: means for receiving said encrypted payment data from said purchaser smart-card; means for decrypting said encrypted payment data received from said purchaser smart-card to obtain payment for the requested goods or services; means for generating acknowledgement data describing the obtained payment; and means for outputting said acknowledgement data for transmission to said vendor smart-card; and wherein said vendor smart-card further comprises means for receiving said acknowledgement data from said 20 vendor registry smart-card.\",\n \"24.A system according to claim 1, further comprising a purchaser terminal associated with the purchaser, the purchaser terminal comprising: (i) means for generating a request for vendor goods or services from said purchaser; (ii) means for interfacing with said purchaser smart-card reader; (iii) means for interfacing with said vendor terminal; and (iv) means for transmitting said purchaser requests to said vendor 30 terminal via said purchaser smart-card.\",\n \"25.A system according to claim 24, wherein data transmitted between said purchaser terminal and said vendor terminal is encrypted before transmission by the respective smart-cards.\",\n \"26.An electronic transaction payment system comprising: a vendor terminal associated with a vendor who provides goods or services to a purchaser; a vendor smart-card; a vendor smart-card reader for transmitting data to and receiving data from said vendor smart-card; a purchaser smart-card; and a purchaser smart-card reader for transmitting data to and receiving data from said purchaser smart-card; wherein said vendor terminal comprises: (i) means for processing requests for vendor goods or services from said purchaser; (ii) means for generating cost data identifying the cost of requested goods or services; (iii) means for interfacing with said vendor smart-card reader; (iv) means for interfacing with said purchaser smart-card reader; and (v) means for transmitting the cost data to said purchaser smart-card via said purchaser smart-card interface; wherein said purchaser smart-card comprises: (i) means for receiving said cost data from said vendor terminal; (ii) means for digitally signing payment data to be transmitted to said vendor smart-card; (iii) means for outputting the signed payment data for transmission to said vendor smart-card; wherein said vendor smart-card comprises: (i) means for receiving said signed payment data from said purchaser smart-card to obtain payment for the requested goods or services; (ii) means for reading the digital signature applied to the payment data to establish the origin of the payment; (iii) means for generating receipt data describing the goods or services and the payment obtained from said payment data; (iv) means for digitally signing said receipt data; and (v) means for outputting 10 the signed receipt data for transmission to said purchaser smart-card, wherein said purchaser smart-card further comprises: (iv) means for receiving said signed receipt data; and (v) means for reading the digital signature applied to the receipt data to establish the origin of the receipt.\",\n \"27.An electronic transaction payment system comprising: a vendor terminal associated with a vendor who provides goods or services to a purchaser; a vendor smart-card; a vendor smart-card reader for transmitting data to and receiving data from said vendor smart-card; a purchaser smart-card; and a purchaser smart-card reader for transmitting data to and receiving data from said purchaser smart-card; wherein said vendor terminal comprises: (i) means for processing requests for vendor goods or services from said purchaser; (ii) means for generating cost data identifying the cost of requested goods or services; (iii) means for interfacing with said vendor smart-card reader; (iv) means for interfacing with said purchaser smart-card; and (v) means for transmitting the cost data to said purchaser smart-card via said purchaser smart-card reader interface; wherein said purchaser smart-card comprises: (i) means for receiving said cost data from said vendor terminal; (ii) means for generating a request for data describing the vendor; and (iii) means for outputting the request for data describing the vendor for transmission 10 to said vendor smart-card; wherein said vendor smart-card comprises: (i) means for receiving said request for data describing the vendor from said purchaser smart-card; (ii) means for generating data describing the vendor in accordance with said request; and (iii) means for outputting the data describing the vendor for transmission to said purchaser smart-card; wherein said purchaser smart-card further comprises: (iv) means for receiving said data describing the vendor from said vendor smart-card; (v) means for determining whether said data describing the vendor conforms to a predetermined condition; (vi) means for, if said data describing the vendor conforms to said predetermined condition, generating payment data to be transmitted to said vendor smart-card; and (vii) means for outputting the payment data for transmission to said vendor smart-card; and wherein said vendor smart-card further comprises: (iv) means for receiving said payment data from said purchaser smart-card to obtain payment for requested goods or services.\",\n \"28.An electronic transaction payment system comprising: a vendor terminal associated with a vendor who provides goods or services to a purchaser; a vendor smart-card; a vendor smart-card reader for transmitting data to and receiving data from said vendor smart-card; a purchaser smart-card; and a purchaser smart-card reader for transmitting data to and receiving data from said purchaser smart-card; wherein said vendor terminal comprises: (i) means for processing requests for vendor goods or services from said purchaser; (ii) means for generating cost data identifying the cost of requested goods or services; (iii) means for interfacing with said vendor smart-card reader; (iv) means for interfacing with said purchaser smart-card reader; and (v) means for transmitting the 20 cost data to said purchaser smart-card via said purchaser smart-card reader interface; wherein said purchaser smart-card comprises: (i) means for receiving said cost data from said vendor terminal; wherein said vendor smart-card comprises: (i) means for generating a request for data describing the purchaser; and (ii) means for outputting the request for data describing the purchaser for transmission to said purchaser smart-card; wherein said purchaser smart-card further comprises: (ii) means for receiving said request for data describing the purchaser from said vendor smart-card; (iii) means for generating data describing the purchaser in accordance with said request; and (iv) means for outputting the data describing the purchaser for transmission to said vendor smart-card; wherein said vendor smart-card further comprises: (iii) means for receiving said data describing the 10 purchaser from said purchaser smart-card; (iv) means for determining whether said data describing the purchaser conforms to a predetermined condition; (v) means for, if said data describing the purchaser conforms to said 15 predetermined condition, generating acceptance data to be transmitted to said purchaser smart-card; and (vi) means for outputting the acceptance data for transmission to the purchaser smart-card; and wherein said purchaser smart-card further comprises: (v) means for receiving said acceptance data from said vendor smart-card; (vi) means for generating payment data to be transmitted to said vendor smart-card; and (vii) means for outputting the payment data for transmission to said vendor smart-card; and wherein said vendor smart-card further comprises: (viii) means for receiving said payment data from said purchaser smart-card to obtain payment for the requested goods or services.\",\n \"29.An electronic transaction payment system comprising: a vendor terminal associated with a vendor who provides goods or services to a purchaser; a vendor smart-card; a vendor smart-card reader for transmitting data to and receiving data from said vendor smart-card; a purchaser smart-card; and a purchaser smart-card reader for transmitting data to and receiving data from said purchaser smart-card; wherein said vendor terminal comprises: (i) means for processing requests for vendor goods or services from said purchaser; (ii) means for generating cost data identifying the cost of requested goods or services; (iii) means for interfacing with said vendor smart-card 15 reader; (iv) means for interfacing with said purchaser smart-card; and (v) means for transmitting the cost data to said purchaser smart-card via said purchaser smart-card reader interface; wherein said purchaser smart-card comprises: (i) means for receiving said cost data from said vendor terminal; (ii) means for generating a request for data describing the vendor; and (iii) means for outputting the request for data describing the vendor for transmission to said vendor smart-card; wherein said vendor smart-card comprises: (i) means for receiving said request for data describing the vendor from said purchaser smart-card; (ii) means for receiving data describing the vendor in accordance with said request; and (iii) means for outputting the data describing the vendor for transmission to said purchaser smart-card; wherein said purchaser smart-card further comprises: (iv) means for receiving said data describing the vendor from said vendor smart-card; (v) means for outputting said data describing said vendor to the purchaser; (vi) means for receiving an indication whether or not said data describing the vendor is acceptable to the purchaser; and (vii) means for, if said data describing the vendor is acceptable, causing payment data to be transmitted to said vendor smart-card; and wherein said vendor smart-card further comprises: (iv) means for receiving said payment data to obtain payment for the requested goods or services.\",\n \"30.An electronic transaction payment system comprising: a vendor terminal associated with a vendor who provides goods or services to a purchaser; a vendor smart-card; a vendor smart-card reader for transmitting data to and receiving data from said vendor smart-card; a purchaser smart-card; and a purchaser smart-card reader for transmitting data to and receiving data from said purchaser smart-card; wherein said vendor terminal comprises: (i) means for processing requests for vendor goods or services from said purchaser; (ii) means for generating cost data identifying the cost of requested goods or services; (iii) means for interfacing with said vendor smart-card reader; (iv) means for interfacing with said purchaser smart-card reader; and (v) means for transmitting the cost data to said purchaser smart-card via said purchaser smart-card reader interface; wherein said purchaser smart-card comprises: (i) means for receiving said cost data from said vendor terminal; wherein said vendor smart-card comprises: (i) means for generating a request for data describing the purchaser; and (ii) means for outputting the request for data describing the purchaser for transmission to said purchaser smart-card; wherein said purchaser smart-card further comprises: (ii) means for receiving said request for data describing the purchaser from said vendor smart-card; (iii) means for receiving data describing the purchaser in accordance with said request; and (iv) means for outputting the data describing the purchaser for transmission to said vendor smart-card; wherein said vendor smart-card further comprises: (iii) means for receiving said data describing the purchaser from said purchaser smart-card; (iv) means for outputting said data describing the purchaser to the vendor; (v) means for receiving an indication whether or not said data describing the purchaser is acceptable to the vendor; (vi) means for, if said data describing the purchaser is acceptable, generating acceptance data to be transmitted to said purchaser smart-card; and (vii) means for outputting the acceptance data for transmission to the purchaser smart-card; and wherein said purchaser smart-card further comprises: (v) means for receiving said acceptance data from said vendor smart-card; (vi) means for generating payment data to be transmitted to said vendor smart-card; and (vii) means for outputting the payment data for transmission to said vendor smart-card; and wherein said vendor smart-card further comprises: (viii) means for receiving said payment data from said purchaser smart-card to obtain payment for the requested goods or services.\",\n \"31.A smart-card programmed to be able to function as a purchaser smart-card according to claim 1.32.A smart-card programmed to be able to function as a vendor smart-card according to claim 1.33.A smart-card programmed to be able to function as a registry smart-card according to claim 13.34.A smart-card comprises: means for interfacing with a card reader; means for receiving details of a new transaction from an external terminal via said interfacing means and said card reader; means for generating a unique session ID for the new transaction; means for storing the generated session ID; means for receiving message data relating to the new transaction from the external terminal via said card reader and said interfacing means; means for tagging the received message data by appending the stored session ID to the message data; and means for outputting the tagged message data to said external terminal via said interface means and said card reader.\",\n \"35.A smart-card according to claim 34, further comprising means for encrypting said tagged message data.\",\n \"36.A smart-card according to claim 34, further comprising means for receiving a session ID generated by another smart-card involved in the new transaction and wherein said tagging means is operable to append the generated session ID and the received session ID to the message data.\",\n \"37.A smart-card according to claim 34, further comprising a money store for storing electronic money and wherein said tagging means is operable to tag said electronic money with said generated session ID and to output the tagged electronic money back to said external terminal via said interface means in said card reader.\",\n \"38.A smart-card according to claim 37, comprising means for storing remote account data and wherein said smart-card is operable to receive an instruction identifying whether or not payment for a current transaction is to be made from electronic money stored on the card or from the remote account and wherein said tagging means is operable to tag said electronic money with said session ID if said instruction identifies that the payment should be made from the card and is operable to tag the stored account data if said instruction indicates that the payment is to be made from the remote account.\",\n \"39.A smart-card according to claim 34, further comprising means for storing user validation data; means for receiving instructions to output user validation data; and wherein said tagging means is operable to tag the user validation data with the session ID prior to outputting the tagged validation data to said external terminal via said interface means and said card reader.\",\n \"40.A smart-card according to claim 34, further comprising a transaction log store for storing details of data transmitted during each new transaction.\",\n \"41.A smart-card comprising: means for interfacing with a card reader; a card controller operable for receiving instructions from an external terminal via said interface means and said card reader; means for storing electronic money; means for storing account data relating to a remote account; wherein said card controller is operable to either output electronic money from said money store or to output account details from said account storing means in dependence upon instructions received from an external terminal via said interfacing means and said card reader.\",\n \"42.A smart-card according to claim 41, wherein said card controller is operable to tag the electronic money or the account data with a unique identifier relating to a current transaction.\",\n \"43.An electronic transaction system comprising a vendor 15 terminal, a purchaser terminal and a third party registry for providing validation of a purchaser associated with the purchaser terminal and/or validation of a vendor associated with the vendor terminal.\",\n \"44.A signal carrying processor implementable instructions for causing a smart-card to become programmed as a smart-card according to claim 31.45.A storage medium storing processor implementable instructions for causing a smart-card to become programmed as a smart-card according to claim 31.46.A computer terminal programmed to function as a vendor terminal according to claim 1.47.A computer terminal programmed to function as a purchaser terminal according to claim 24.48.An electronic transaction payment characterized by the step of using an electronic transaction payment system according to claim 1.\"\n ],\n [\n \"This invention relates to a payment system and in particular but not exclusively to a smart-card payment system wherein an entire purchase operation is explicitly linked as a single transaction and a variable level of user validation is provided.\",\n \"Existing payment systems suffer from a number of drawbacks.\",\n \"The use of cash for example is limited by the requirement for the actual transfer of possession of physical tokens and by the fact that if lost or stolen, cash is usable without restriction by a finder or thief.\",\n \"The use of cheques goes some way to overcome the security worries of carrying cash but still requires the transfer of a physical token.\",\n \"Cheques also introduce the problem of “clearing time”, that is, time taken by banks to actually transfer money from the cheque writer's account to the cheque receiver's account.\",\n \"In addition, there is a possibility with a cheque that a bank may refuse to honour a cheque written by an account holder because the account holder has insufficient funds in their account.\",\n \"In this circumstance, the receiver of the cheque has then transferred ownership of goods or provided a service and has received no payment for those goods or that service.\",\n \"For ordering goods by telephone or using the internet, cash is totally unsuitable and cheques are very inconvenient because to pay by cheque for goods ordered by telephone or over the internet requires the order placed using the telephone or the internet to be confirmed in writing accompanied by a cheque for payment.\",\n \"More suitable for use with internet or telephone orders are credit and debit cards.\",\n \"When using a credit card, payment for the goods is actually made to the retailer by a credit card company which allows the credit card user to spend up to a predetermined borrowing limit for purchases.\",\n \"Purchases made with a credit card must be paid for to the credit card company within a predetermined time limit or interest on the money borrowed becomes payable.\",\n \"Debit cards on the other hand provide for an electronic transfer of funds direct from the card holder's bank account to the retailer's bank account.\",\n \"Credit and debit cards also suffer from the problems of “clearing time”.\",\n \"Both credit and debit cards use conventional magnetic stripe card technology.\",\n \"Both payment systems also suffer from processing charges inherent in the system.\",\n \"These processing charges are typically levied on the vendor and may be in the range of one to four percent of the sale transaction.\",\n \"Sometimes all or part of this cost is passed onto the purchaser.\",\n \"This transaction cost has a minimum size which is set by the banks and credit card companies operating the cards.\",\n \"As a result, it is economically disadvantageous to process small value transactions using credit and debit cards.\",\n \"Credit and debit cards do however have the advantage for telephone and internet transactions that there is no requirement for the transfer of physical tokens.\",\n \"They can therefore be used to make purchases or to reserve restaurant tables for example, without the need for postal or personal transfer of payment.\",\n \"The use of a credit card to make a purchase provides the purchaser with extra protection in that the credit card company must cover the cost of goods paid for on the card, over a certain minimum value, which are not delivered or with which there are certain other problems.\",\n \"In addition, the credit card operator pays the vendor regardless of whether or not the purchaser ever repays that money to the credit card company.\",\n \"Thus both vendor and purchaser benefit from the use of a credit card.\",\n \"However, for telephone and internet transactions, which are defined by the credit card operator as “card holder not present” transactions, the risk of loss rests with the retailer not the card operator.\",\n \"Thus, it becomes less beneficial for a vendor to accept credit cards as payment for telephone and internet orders because they must pay the handling fee and they receive no protection.\",\n \"The so-called electronic purse (e.g.\",\n \"the Mondex Electronic Purse System) has been developed to eliminate the problems of carrying actual cash and of the minimum processing charges for small debit or credit card transactions.\",\n \"The electronic purse makes use of smart-card or chip-card technology.\",\n \"To use the electronic purse, the user must either load cash onto the smart-card electronic purse at a vending machine or at the desk of a load agent such as a Post Office, or the user must transfer funds from their bank account or credit card to the smart-card electronic purse.\",\n \"The purse may then be used to make purchases in the real world, over the internet or over the telephone, subject to the telephone being with an appropriate smart-card reader.\",\n \"From a transaction processing point of view, full clearing would only be required when loading the purse, which would normally be a medium to large value transaction and therefore acceptable.\",\n \"Payments could be batched into daily totals and cleared to the retailers as single daily transactions, again being of reasonable size.\",\n \"There would be no requirement to account for every transaction back to the card holder, all that would happen would be that payment would be made out of the cash float created when taking cash to load the electronic purse.\",\n \"Thus from a bank's point of view, the electronic purse solved a major problem.\",\n \"However, from the card holder's point of view the electronic purse is less successful and all open electronic purse trials have tended towards failure in terms of take up.\",\n \"Among the reasons for the poor take up are: poor education of the card holders, small scale trials leading to purse being usable at only a few locations, lock-in of card holder money to the card and places it could be used, no interest on prepayment, requirement for a bank account to back the electronic purse and fear over security and transaction accountability.\",\n \"The requirement for a bank account to back the electronic purse is a major failing of the system.\",\n \"This is because many people who could best benefit from an electronic purse (or other prepaid mechanisms) are those people who do not have bank accounts and who are therefore automatically excluded from using cheques, debit cards or credit cards.\",\n \"Approximately twenty percent of the UK population are disallowed by the banks from having a bank account.\",\n \"It should be noted that the Mondex Electronic Purse System was specifically developed to allow use by persons not having a bank account, however the operators still insisted on a user having a bank account before a card would be issued.\",\n \"Regarding fear over security and transaction accountability, the problem is one of how much the card holder trusts the retailer.\",\n \"For example, how does the card holder know how much credit is stored on his or her card and how do they know how much is removed?\",\n \"For internet transactions, this problem grows to incorporate issues such as: how to identify how much is being deducted from the card, how to identify the other party to the transaction, how to identify the location of the other party to the transaction, whether goods will actually be shipped, whether the payment made will actually go to the vendor, whether the vendor will recognise the payment as being against the goods purchased, whether the payment could be fraudulently redirected and concerns over who will be able to see the delivery address specified.\",\n \"The existing electronic purse schemes use so-called smart-cards or chip-cards.\",\n \"These are cards having a microprocessor and memory mounted onto or into a card, with an interface provided for the microprocessor to be powered and interfaced with by a reader.\",\n \"The standards for smart-cards where the interface is by means of physical connection terminals are set out in ISO 7816.The standards for smart-cards where the interface is by means of radio signals (including supply of power) are set out in ISO 14443.In an attempt to address some of the above problems, a number of banks and financial card operators including Mastercard and Visa have agreed a smart-card standard for credit and debit services such that a retailer could have a common terminal to support all smart credit and debit cards in a similar manner to the current situation with magnetic stripe cards.\",\n \"This standard is known as EMV (Europay Mastercard Visa) and it applies to credit and debit services on smart-cards as well as the terminals to support them.\",\n \"An electronic purse system based on the EMV specifications is the common electronic purse specifications (CEPS) with which the European Standard Organisation (CEN) has been involved.\",\n \"CEPS requires a personalised system and additional hardware to identify the cardholder in order for him or her to digitally sign and certify all CEPS transactions.\",\n \"Further, for use in telephone or internet transactions, there is a requirement in CEPS for digital signatures.\",\n \"There is currently no worldwide or European standard for producing digital signatures.\",\n \"Such digital signatures require a public key infrastructure administered by a certification authority which acts as a library or directory of public keys.\",\n \"In addition, when it supplies a public key it adds a certificate signed with its own private key such that the recipient can then validate that the public key information is genuine within Europe, the various public key infrastructures that currently exist are not compatible and therefore such digital signatures are not interoperable across services and countries.\",\n \"Thus the full implementation of CEPS will be delayed until such standardisation can be completed.\",\n \"In summary therefore, cash and cheques require the exchange of physical tokens and are therefore totally unsuitable for real time trading by telephone or over the internet.\",\n \"Credit and debit cards and cheques suffer from clearing time delays.\",\n \"Credit and debit cards are also unsuitable for small value transactions due to the processing charges included.\",\n \"Electronic purses have been unpopular due to the “tied-in” nature of the funds and the small number of places where trials have allowed use of the purses.\",\n \"Credit and debit cards and electronic purses have the additional problem for internet and telephone transactions of requiring trust between purchaser and vendor.\",\n \"CEPs has the drawback of an awkward and non-standard security system.\",\n \"Aspects of the present invention relate to a smart-card payment system wherein: provision is made for payment by means of prepaid value stored on a card held by the purchaser (the so-called electronic purse); provision is made for payment by means of credit by a third party on behalf of the purchaser and accessed by means of a smart-card held by the purchaser (the so-called credit card facility); provision is made for payment by means of prepaid value stored on behalf of the purchaser and accessed by means of a smart-card held by the purchaser (the so-called debit card facility); provision is made for different levels of validation between the purchaser and the vendor including third party validation if required; the provision of a third party registry to permit the movement of value or funds between the registry and the smart-card; payment may be anonymous or personalised; the purchaser may use either of the electronic purse and debit card facilities without having to pass any means test, credit check or hold a bank account; and a system in which an entire purchase operation is inextricably linked as a single transaction and variable levels of user (purchaser and/or vendor) validation is provided.\",\n \"The present invention seeks to address difficulties of the prior art.\",\n \"According to one aspect, there is provided an electronic transaction payment system comprising a vendor terminal associated with a vendor who provides goods or services to a purchaser; a vendor smart-card; a vendor smart-card reader for transmitting data to and receiving data from the vendor smart-card; a purchaser smart-card reader for transmitting data to and receiving data from a purchaser smart-card.\",\n \"The vendor terminal comprises means for processing requests for vendor goods or services from the purchaser, means for generating cost data identifying the cost of requested goods or services and means for transmitting the cost data to the purchaser smart-card.\",\n \"The purchaser smart-card includes means for receiving the cost data from the vendor terminal and means for encrypting the payment data for transmission back to the vendor smart-card.\",\n \"The vendor smart-card also includes means for receiving the encrypted payment data and means for decrypting the payment data to obtain payment for the requested goods or services.\",\n \"Preferably, all communications relating to the purchase of the goods pass from the purchaser's smart-card to the vendor and are encrypted with an encryption key specific to the current transaction.\",\n \"In this way, payment of the goods and/or services can be inextricably linked to the transaction itself.\",\n \"The invention can be applied for electronic trading on, for example, the Internet and can be used at traditional points of sale.\",\n \"In a preferred embodiment, a third party registry is provided to provide guarantees for the purchaser and/or the vendor if required.\",\n \"The third party registry may provide additional support as requested and to a level requested by the vendor or purchaser, to ensure trust and confidence in the trade.\",\n \"Either party in the transaction may choose to validate the credentials of the other party by making a reference to the third party registry.\",\n \"In this case, the registry will provide the information and may levy an appropriate fee.\",\n \"Preferred embodiments of the present invention will now be described by way of example only with reference to the accompanying drawings, in which: FIG.\",\n \"1 shows an internet enabled computing environment in which the present invention may operate; FIG.\",\n \"2 is a block diagram showing the main functional elements of a vendor terminal and a purchaser terminal shown in FIG.\",\n \"1; FIG.\",\n \"3 is a block diagram showing the main functional elements of a registry terminal shown in FIG.\",\n \"1; FIG.\",\n \"4 is a block diagram showing the main functional elements of a smart-card shown in FIG.\",\n \"1; FIG.\",\n \"5 is a flow chart showing the main processing steps performed by the terminals in FIG.\",\n \"1 to process a transaction; FIG.\",\n \"6 is a flow diagram showing in more detail the processing steps involved in a product selection and price agreement step which forms part of the processing steps shown in FIG.\",\n \"5; FIG.\",\n \"7 which comprises FIGS.\",\n \"7A and 7B is a flow diagram showing in more detail the processing steps involved in a secure link set-up step which forms part of the processing steps shown in FIG.\",\n \"5; FIG.\",\n \"8 is a flow diagram showing in more detail the processing steps involved in a purchaser validation step which forms part of the processing steps shown in FIG.\",\n \"5; FIG.\",\n \"9 is a flow diagram showing in more detail the processing steps involved in a vendor validation step which forms part of the processing steps shown in FIG.\",\n \"5; FIG.\",\n \"10 is a flow diagram showing in more detail the processing steps involved in a delivery confirmation step which forms part of the processing steps shown in FIG.\",\n \"5; FIG.\",\n \"11 which comprises FIGS.\",\n \"11A and 11B is a flow diagram showing in more detail the processing steps involved in a payment and receipting step which forms part of the processing steps shown in FIG.\",\n \"5; FIG.\",\n \"12 which comprises FIGS.\",\n \"12A and 12B is a flow diagram showing in more detail the processing steps involved in a purchaser validation step which forms part of the processing steps shown in FIG.\",\n \"5 according to an alternative embodiment; FIG.\",\n \"13 is a flow diagram showing in more detail the processing steps involved in a purchaser validation step which forms part of the processing steps shown in FIG.\",\n \"5 according to a further alternative embodiment; FIG.\",\n \"14 which comprises FIGS.\",\n \"14A and 14B is a flow diagram showing in more detail the processing steps involved in a vendor validation step which forms part of the processing steps shown in FIG.\",\n \"5 according to another alternative embodiment; FIG.\",\n \"15 is a flow diagram showing in more detail the processing steps involved in a payment and receipting step which forms part of the processing steps shown in FIG.\",\n \"5 according to another alternative embodiment; FIG.\",\n \"16 is a flow diagram showing in more detail the processing steps involved in a payment and receipting step which forms part of the processing steps shown in FIG.\",\n \"5 according to another alternative embodiment; FIG.\",\n \"17 is a flow diagram showing in more detail the processing steps involved in a payment and receipting step which forms part of the processing steps shown in FIG.\",\n \"5 according to a further alternative embodiment; FIG.\",\n \"18 which comprises FIGS.\",\n \"18A and 18B is a flow diagram showing the main processing steps required to convert currency on an account card; FIG.\",\n \"19 shows an internet enabled face-to-face retail environment in which the present invention may operate; FIG.\",\n \"20 is a block diagram showing the main functional elements of a retailer terminal shown in FIG.\",\n \"19.OVERVIEW In the following description it is assumed that the purchaser's terminal is already connected to the internet, as is the vendor's terminal.\",\n \"Also, using his or her terminal, the purchaser has browsed the vendor's website and has selected a product or products that he or she wishes to purchase.\",\n \"Referring to FIG.\",\n \"1, the internet enabled computing environment 1 in which the payment system of the present embodiment is operated, comprises a purchaser terminal 5, a vendor terminal 21 and a registry terminal 25 which communicate electronically with one another via a network such as the internet 19.Attached to the purchaser terminal 5 is a smart-card slot 7p into which a purchaser smart-card 3 may be inserted.\",\n \"In this embodiment, the purchaser smart-card 3 must be inserted into the smart-card slot 7p in order for any transaction to be made with a vendor.\",\n \"Also attached to purchaser terminal 5 is a monitor 9 for displaying data to a user, a keyboard 11 and a mouse 13 which provide a user interface to the purchaser terminal 5 and a modem 15 for communicating via the internet 19.An electrical, electromagnetic or optical signal 17 is transmitted from and received by modem 15 in order to facilitate communication via the internet 19.As shown in FIG.\",\n \"1, the vendor terminal 21 also includes a monitor 9, a keyboard 11, a mouse 13, a modem 15 and a smart-card slot 7v for receiving a vendor smart-card 23.Attached to the registry terminal 25 is a registry smart-card server 27 having a plurality of registry account smart-cards permanently inserted in card slots 7r therein.\",\n \"In this embodiment, there is typically one registry account logical smart-card for each user of the payment system, although in practice many such logical smart-cards will be grouped onto one master smart-card which will substantially reduce the smart-card and smart-card reader requirement at the registry.\",\n \"Each registry account is represented by a logical record on a master account card.\",\n \"The number of master account cards will be equal to the number of accounts divided by the number of logical account records on each card.\",\n \"Since the information on the card need only contain an account number and balance, with further information held in a connected hard disk at the registry 25, there could be as many as five thousand logical account records per master card.\",\n \"In this embodiment, every registry account is represented by a disk based record at the registry 25.As and when a registry account is to be accessed for whatever reason, key information is copied to a spare logical account slot in a master card, so that card to card working may take place.\",\n \"The number of master cards required will be just greater than the number of parallel transactions required to provide adequate performance.\",\n \"After a transaction is completed, the disk record is updated.\",\n \"The card record is retained until the logical account slot is required.\",\n \"This provides a form of caching to provide better performance for those actively using their smart-cards and registry.\",\n \"Typically, a retailer's account will be more active than a purchaser's account.\",\n \"FIG.\",\n \"2 shows the functional elements of the purchaser terminal 5 and the vendor terminal 21.As shown, these terminals include a central processing unit (CPU) 41 which carries out processing operations in accordance with instructions stored in a working memory 43 and in accordance with user input received either via the keyboard and mouse interface 49 or the smart-card reader 53.The terminals also include a monitor interface 51 for interfacing to the display, a modem interface for interfacing with the modem 15 and a non-volatile software store 47 (such as a hard disk) which stores the processing instructions used to control the operation of the CPU 41 together with other data used by the terminals.\",\n \"As shown, these components are connected together via a bus 45.FIG.\",\n \"3 shows the functional elements of the registry terminal 25.As shown, the registry terminal 25 also includes a CPU 41, a working memory 43, a software store 47, a monitor interface 51 and a modem interface 55 which are all connected together by a bus 45.In addition, the smart-card server 27 is connected to the registry terminal 25 via a smart-card server interface 57.FIG.\",\n \"4 shows the functional elements of a smart-card 3,23 used in this embodiment.\",\n \"As shown, the smart-card 3,23 includes a card controller 62 which carries out processing operations stored in a processing instructions store 64 in accordance with instructions received from the purchaser terminal 5, vendor terminal 21 or the registry terminal 25 via a card reader interface 66.Coupled to the card controller 62 is a money store 68, which holds data representing the amounts of money held on the smart-card 3,23.The smart-card 3,23 also includes a user validation store 70 which stores information about the smart-card owner such as the owner's name, age, address etc.\",\n \"When a smart-card 3,23 is used to make a transaction, a single unique session ID is generated by a session ID generator 72 on the smart-card 3,23.The generated session ID is encrypted by an encryption engine 74 also on the smart-card 3,23 and then stored in a session ID store 76.As will be described in further detail later, this session ID is appended to all subsequent messages related to the current transaction by the card controller 62, and the card controller 62 is also operable to compare the session ID of received messages to the stored session ID to check that the message belongs to the current transaction.\",\n \"As shown in FIG.\",\n \"4, the smart-card 3,23 further comprises a key store 78 for storing data needed for encryption and decryption, such as public and private keys, which will be described in more detail later.\",\n \"The key store 78 is coupled to the encryption engine 74, so that messages relating to a transaction can be encrypted by the encryption engine 74 using the stored encryption key data, before transmission back to the terminal via the card reader interface.\",\n \"Also coupled to the card controller 62 is a transaction log store 80 for storing data representing each transaction made using the smart-card 3,23 and a registry data store 82 for storing details of the user account held at the registry.\",\n \"Referring now to FIG.\",\n \"5, there are shown the principal functional steps which must be performed to complete a purchase transaction using the payment system of the present embodiment.\",\n \"As shown, at step S4-1, the user, having selected a desired product already, provides input indicating a product selection to the purchaser terminal 5.The purchaser terminal 5 then communicates with the vendor terminal 21 by transmitting signals 17 between the modems 55 attached to each of the purchaser terminal 5 and the vendor terminal 21 via the internet 19.The purchaser terminal 5 transmits the product selection to the vendor terminal 21, which then returns price and delivery terms to the purchaser 5.The terms are then displayed by the purchaser terminal 5 on monitor 9 and the purchaser terminal 5 then awaits an input from the purchaser to indicate approval or disapproval of those terms via the keyboard 11 or mouse 13.The inputted approval or disapproval is then communicated by the purchaser terminal 5 to the vendor terminal 21 via the internet.\",\n \"Next, at step S4-3, the purchaser smart-card 3 inserted in the purchaser terminal 5 and vendor smart-card 23 inserted in the vendor terminal 21 create and exchange secure encryption keys (which are stored in the respective key stores 78 of the purchaser and vendor smart-cards 3,23) to be used throughout the payment session and to be used as the basis for the secure exchange of information using data encryption between terminals.\",\n \"Before payment is made, however, in this embodiment the smart-cards 3,23 return control to the terminals so that user validation can be performed.\",\n \"This is initiated at step S4-5, where the vendor terminal 21 displays on monitor 9 a request for user input to indicate whether validation of the purchaser should be undertaken.\",\n \"Such input is received via the keyboard 11 or mouse 13.If validation is to be undertaken, processing proceeds to step S4-7 where validation is performed.\",\n \"To perform validation, as will be described in more detail below, the vendor terminal 21 communicates with purchaser terminal 5 or the registry 25 via the internet 19 to request identifier data about the purchaser so that certain pre-specified levels of knowledge about the purchaser are obtained.\",\n \"Following purchaser validation, or if purchaser validation is not to be undertaken, the processing proceeds to step S4-9, where the purchaser's terminal 5 displays on monitor 9 a request for user input to indicate whether validation of the vendor should be undertaken.\",\n \"If vendor validation is required, then at step S4-11, validation of the vendor is performed, which is a similar process to that described above for purchaser validation.\",\n \"Following such validation, or if validation of the vendor is not required, processing continues at step S4-13.At this step, it is decided whether or not delivery of the product will be required.\",\n \"To determine whether delivery is required, the vendor terminal 21 analyses the delivery terms agreed at step S4-1.From these terms it will be clear whether anything is to be delivered.\",\n \"It is to be understood that delivery may include electronic transfer of data as well as postal delivery of physical items.\",\n \"If delivery is required, then the delivery data is confirmed at step S4-15 where the purchaser terminal transmits the relevant delivery address (which may be a postal address, an e-mail address or an FTP (file transfer protocol) server address for example) to the vendor terminal 21.Following this, or in the event that delivery is not required, control is passed back to the smart-cards 3,23 where payment and receipting is performed at step S4-17.In this step, the purchaser terminal 5 causes the appropriate value to transfer from the money store 68 of the purchaser smart-card 3 to the money store 68 of the vendor smart-card 23, during which a receipt message is transferred from the vendor smart-card 23 to the purchaser smart-card 3.An overview of the smart-card payment system of this embodiment has been given above.\",\n \"The system offers a number of advantages, including: i) the provision of a single session ID which links all messages in a transaction including the terms, user validation and payment, thereby inextricably linking the payment to the transaction; ii) the provision of different levels of user (purchaser and/or vendor) validation; and iii) the provision of a third party registry for validation of one or more of the parties to a transaction if required.\",\n \"These and other advantages of the payment system of the present embodiment will become apparent from the following more detailed description.\",\n \"Product Selection FIG.\",\n \"6 shows an expansion of step S4-1 in which the purchaser terminal 5 transfers data describing a product selection to the vendor terminal 23 and the purchaser and vendor terminals 5, 23 communicate to establish the price and delivery terms for the transaction.\",\n \"In FIG.\",\n \"6, the steps undertaken by the purchaser's terminal are shown to the left hand side of the central dotted line and those steps undertaken by the vendor's terminal are shown on the right hand side of the central dotted line.\",\n \"At step S5-1, the purchaser terminal 5 receives product selection input from the user via the keyboard 11 or mouse 13, and at step S5-3 product selection data is transmitted to the vendor terminal 21 by transmitting signals 17 from the modem 55 connected to the purchaser terminal 5 to the modem 55 connected to the vendor terminal 21 via the internet 19.The product selection data is received by the vendor terminal 21 at step S5-5.At step S5-7, the vendor terminal 21 compares the product selection data to a product database to determine the price of the product and at step S5-11 the price and any necessary delivery terms data are transmitted to the purchaser terminal 5.At step S5-13 the purchaser terminal 5 receives the price and delivery terms data, which data is then displayed to a user at step S5-15.The purchaser terminal 5 then receives acceptance input from the user at step S5-17 and at step S5-19 the terminal 5 processes the input from the user to determine whether the user accepted the price and delivery terms.\",\n \"If the price and delivery terms are not accepted, then at step S5-21 the purchaser terminal 5 transmits that the terms are not accepted to the vendor terminal 21 which is received at step S5-23.The transaction process then ends.\",\n \"If the user does accept the price and delivery terms, then at step S5-25 the purchaser terminal 5 transmits that the terms have been accepted and at step S5-27 the vendor terminal 21 receives that the terms have been accepted.\",\n \"Following this, the selection of product and agreement of price and delivery terms is complete and referring again to FIG.\",\n \"5 processing then continues to step S4-3.Establish Secure Link The processing of step S4-3 is shown in more detail in FIG.\",\n \"7.At step S6-1, the vendor terminal 21 transmits a request for the purchaser to identify how payment is to be made.\",\n \"At step S6-3, that request is received at the purchaser terminal 5 and at step S6-5 the purchaser terminal 5 transmits that the payment system of the present embodiment is to be used, ie payment using the smart-cards 3,23.This is received at step S6-7 by the vendor terminal 21.Once it has been established that the payment system of the current embodiment is to be used, control passes to the smart-cards 3,23 which establish a secure session between themselves.\",\n \"This is achieved, in this embodiment, using so called public key encryption where each party transmits to the other a public key with which data should be encrypted.\",\n \"It is only possible to decrypt the encrypted data using a private key which is retained by each party.\",\n \"The present embodiment uses a modified public key encryption scheme which allows a new, unique session ID to be used for each transaction in connection with the asymmetric key pair.\",\n \"The setting up of the secure session begins at step S6-9 where the vendor smart-card 23 transmits its public key from its key store 78 to the purchaser smart-card 3.The purchaser smart-card 3 receives the public key from the vendor smart-card 23 at step S6-11 and stores it in its key store 78.Next, at step S6-13, the purchaser smart-card 3 generates, using the session ID generator 72, a purchaser session ID which it appends to all communications transmitted to the vendor smart-card 25 during the current session.\",\n \"In the present embodiment, the purchaser session ID takes the form of a block of data made up of a time stamp, a random number, a terminal ID and a card ID, all encrypted using the purchaser smart-card's public key (so that only the purchaser smart-card 3 can decrypt the purchaser session ID).\",\n \"Then at step S6-15, the purchaser smart-card 3 encrypts the purchaser session ID using the vendor smart-card's public key and transmits this to the vendor smart-card 23, along with its own public key.\",\n \"At step S6-17, the vendor smart-card 23 receives and decrypts the data to recover the purchaser session ID and the purchaser public key, which it stores within the session ID store 76 and the key store 78 respectively of the vendor smart-card 23.Next, at step S6-19, the vendor smart-card 23 generates a vendor session ID using its session ID generator 72.In the present embodiment, the vendor session ID comprises a time stamp, a random number, a terminal ID and a card ID, all encrypted using the vendor smart-card's public key (so that only the vendor smart-card 23 can decrypt the vendor session ID).\",\n \"At step S6-21, the vendor smart-card 23 then encrypts the vendor session ID using the purchaser smart-card's public key and transmits it to the purchaser smart-card 3.At step s6-23, the purchaser smart-card 3 receives and decrypts the data to recover the vendor session ID which it stores in its session ID store 76.At step S6-25, the purchaser smart-card 3 transmits a message to the vendor smart-card 23 stating that the vendor session ID has been received, which message is received by the vendor smart-card 23 at step S6-27.The session ID generated by each of the purchaser smart-card 3 and the vendor smart-card 23 will be different for a given session (because of the time stamp and the random number), but they will be inextricably linked to the one session and the two smart-cards.\",\n \"In the present embodiment, the session IDs are added to every transfer of data passing between the terminals after it has been established that the payment system of the present embodiment is to be used.\",\n \"This will take the form of appending both session IDs to the end of the message and then encrypting the whole with the recipient's public key.\",\n \"By performing the encryption each message is protected from being read by an unauthorised party, and by using the session ID each message is digitally signed to certify that the sender of the data is who they claim to be and that the message is valid.\",\n \"It should be noted that only the purchaser smart-card 3 and the vendor smart-card 23 will be able to: validate the session IDs, encrypt data and decrypt data.\",\n \"The cards will be use their own encryption engines 74 for this purpose, rendering it unnecessary for the terminals to be secure devices.\",\n \"Part of the session will include the transmission of a user input such as acceptance or refusal of validation information supplied and/or delivery data.\",\n \"This will be entered as plain text (i.e.\",\n \"unencrypted and uncoded) by the vendor or purchaser and then passed to the local smart-card for appending of session IDs, encryption and transfer to the other party.\",\n \"Again, it is not necessary for the terminal to be a secure device.\",\n \"Purchaser Validation Referring now to FIG.\",\n \"8, the processing steps to perform the validation of a purchaser (step S4-7) will now be described.\",\n \"First, the vendor or vendor terminal 23 must determine the level of validation required at step S7-1.Note that if it is determined at step S4-5 that no validation is required, then the processing steps shown in FIG.\",\n \"8 will not be undertaken.\",\n \"In this embodiment, the level of validation may be selected manually by the vendor, in which case validation options will be displayed by the vendor terminal 21 on monitor 9 and which will then await an input from the vendor via the keyboard 11 or the mouse 13.Alternatively, the level of validation may be selected by the vendor terminal automatically, in which case the characteristics (eg value, nature etc) of the transaction will be compared to a predetermined set of rules for deciding the level of validation required.\",\n \"Once the level of validation has been determined, the vendor terminal 21 will transmit a request for the purchaser validation at step S7-3 indicating the level of validation required.\",\n \"In the present embodiment, it is assumed that a relatively low level of validation is required.\",\n \"Therefore the validation request is sent to the purchaser terminal 5, not the registry 25.At step S7-5, the purchaser terminal 5 receives the validation request and at step S7-7 transmits the requested validation data (which may either be input by the purchaser or is stored in the validation store 70 of the purchaser smart-card 3).\",\n \"At step S7-9 the vendor terminal 21 receives the validation data and at step S7-11 determines whether the validation data received is acceptable.\",\n \"This is done either automatically by comparing the received validation data with pre-stored data or manually by displaying the validation data to the vendor and awaiting confirmation from the vendor that it is acceptable.\",\n \"If the validation data is unacceptable then processing continues at step S7-13 where the vendor terminal 21 transmits to the purchaser terminal 5 that the validation data provided was unacceptable.\",\n \"This is received at step S7-15 by the purchaser terminal 5 and then the transaction process ends.\",\n \"Alternatively, if the validation data is acceptable, processing continues at step S7-17 where the vendor terminal 21 transmits to the purchaser terminal 5 that the validation data provided was acceptable and at step S7-19 the purchaser terminal 5 receives this.\",\n \"The validation of the purchaser is then complete and, referring again to FIG.\",\n \"5, processing continues at step S4-9.Vendor Validation If it is decided at step S4-9 that validation of the vendor is required, as described above, then the steps shown in FIG.\",\n \"9 are performed (step S4-11).\",\n \"The processing of FIG.\",\n \"9 is substantially the same as the processing of FIG.\",\n \"8, except that it is the purchaser terminal 5 requesting the validation data and the vendor terminal 21 transmitting it.\",\n \"Thus at step S8-1, the purchaser terminal 5 determines the level of validation that is required in substantially the same manner as described above with reference to purchaser validation in FIG.\",\n \"8, following which it transmits a vendor validation request either to the vendor terminal 21 or to the registry 25 at step S8-3.In the present embodiment it is assumed that a relatively low level of validation is required.\",\n \"Therefore the validation request is sent to the vendor terminal 21, not the registry 25.On receiving the vendor validation request at step S8-5, the vendor terminal 21 transmits, at step S8-7, the requested validation data (which may either be input by the vendor or is stored in the validation store 70 of the vendor smart-card 23).\",\n \"Following receipt of the validation data by the purchaser terminal 5 at step S8-9, the purchaser terminal 5 determines at step S8-11 whether the validation data is acceptable.\",\n \"This is done either automatically by comparing the received validation data with pre-stored data or manually by displaying the validation data to the purchaser and awaiting confirmation from the purchaser that it is acceptable.\",\n \"If the validation data is decided to be unacceptable, then at step S8-13 the purchaser terminal 5 transmits to the vendor terminal 21 that the validation data is not acceptable and the vendor terminal 21 receives this at step SB-15 following which the transaction process terminates.\",\n \"If, however, the validation data is determined to be acceptable at step S8-11, then at step S8-17 the purchaser terminal 5 transmits to the vendor terminal 21 that the validation data is acceptable.\",\n \"This acceptance is received by the vendor terminal 21 at step S8-19 following which the validation of the vendor is complete and, referring again to FIG.\",\n \"5, processing continues at step S4-13.Confirm Delivery Data If it is determined at step S4-13 that delivery is required, then the delivery data must be confirmed at step S4-15.This process is shown in more detail in FIG.\",\n \"10.At step S9-1, the vendor terminal 21 requests the delivery data, that is the delivery address (which may be a postal address or an e-mail address, for example) and the name of the person or company to whom the items should be directed.\",\n \"At step S9-3, the purchaser terminal 5 receives the delivery data request and displays on monitor 9, at step S9-5, a request for a user to enter delivery data.\",\n \"At step S9-7, the purchaser terminal receives the delivery data input from the user via the keyboard 11 or the mouse 13 and at step S9-9 transmits the delivery data to the vendor terminal 21.The vendor terminal 21 then receives the delivery data at step S9-11, following which the process of confirming delivery data is complete and, referring again to FIG.\",\n \"4, the processing will continue at step S4-17.Payment and Receipting Performing payment and receipting (step S4-17) will now be described in greater detail with reference to FIG.\",\n \"11.As mentioned above, control of the payment and receipting is carried out in this embodiment by the smart-cards 3,23.Starting at S10-1, the vendor smart-card 23 transmits a request for payment to the purchaser smart-card 3.At step S10-3, the purchaser smart-card 3 receives the request and at step S10-5 transmits the payment data to the vendor smart-card 23.At step S10-7, the vendor smart-card 23 receives the payment data and at step S10-9 transmits a receipt for the payment.\",\n \"At this time, the vendor smart-card 23 has received the payment data but has not accepted the payment described therein.\",\n \"The purchaser smart-card 3 then receives the receipt at step S10-11.At step S10-12, the purchaser smart-card 3 validates the receipt by checking that the purchaser session ID has been appended to the receipt data and that it matches its purchaser session ID stored in the session ID store 76.If the purchaser session ID is not present and correct, then at step S10-13, the purchaser smart-card 23 transmits an error message to the vendor smart-card 23 stating that the session ID is incorrect.\",\n \"This error message is received by the vendor smart-card 23 at step S10-14 following which the transaction process ends.\",\n \"On the other hand, if it is determined at step S10-12 that the purchaser session ID is present and correct, then at step S10-15 the purchaser smart-card 3 transmits an acknowledgement that the receipt has been received.\",\n \"This acknowledgement is received by the vendor smart-card 23 at step S10-16, following which the vendor smart-card 23 validates the acknowledgement by checking that the vendor session ID has been appended to the acknowledgement data and that it matches the vendor session ID stored in the session ID store 76.If the vendor session ID is not present and correct, then at step S10-18 the vendor smart-card 23 generates an error message stating that the vendor session ID is not present and transmits it to the purchaser smart-card 3 which receives the message at step S10-19 following which the transaction process ends.\",\n \"If on the other hand, however, it is determined at step S10-17 that the vendor session ID is present and correct, then processing continues at step S10-20 where the vendor smart-card 23 finalises the order data and accepts the payment data, thereby actually receiving the payment, to complete the transaction.\",\n \"The receipt and acknowledgement are validated by each smart-card 3 and 23 to ensure the session remains intact.\",\n \"The transaction can only be completed if both smart-cards 3 and 23 complete this test successfully.\",\n \"Thus the payment process is inextricably linked to the purchase transaction, the validation of the parties and acceptance of all terms into a single transaction session including user input of validation acceptance and possible delivery details.\",\n \"Once the session has been validated as complete the vendor smart-card 23 transmits, at steps s10-21, a message to the purchaser terminal 5 to indicate that the transaction is complete.\",\n \"At step S10-22 the purchaser smart-card 3 receives the transaction complete message following which the transaction process terminates.\",\n \"Following completion of the transaction, the purchaser terminal 5 and smart-card 3 may then be used to perform other actions unconnected with the purchase.\",\n \"Validation Level Determination As described above, it is necessary to determine at step S7-1 and S8-1 the level of validation of the other party to a transaction that is required.\",\n \"For example, one party may wish to validate the address and other credentials (such as age, legal status or credit worthiness) of the other party and also to be guaranteed that purchased goods will be delivered intact and possibly to a given quality.\",\n \"As discussed above, some of this information, such as name, age or address, is held in the validation store 70 on the smart-card of the user and this information plus further information may be held by the registry 25.Either party may opt for more or less information from the registry 25, and/or a greater or lower level of guarantee.\",\n \"The registry 25 may implement a charging structure to enable users of the payment system to be charged for information held by and requested from the registry 25, or to charge users for storing and providing their data to others.\",\n \"Different levels of payment will be required depending on the amount of validation data held by the registry.\",\n \"In all trades made using the payment system of the present invention, the complete trade, that is selection of goods, agreement to purchase, payment and subsequent expected delivery are based on the honesty of the two parties and their acceptance of each other's bona fides and mutual acceptance of the terms of trading.\",\n \"The payment system does not apply any specific trading rules of its own and it is up to both parties to agree terms that overcome any doubts a party may have regarding the other party and the trade.\",\n \"The payment system provides electronic packaging of a negotiation between the parties such that value does not move from one party to the other until both parties are satisfied with the terms.\",\n \"Thus, each party is able to decide upon the level of trust it has in the other party and if insufficient, to make use of the registry 25 as necessary until satisfied.\",\n \"For example, two parties trading for the first time may both seek the top level of guarantee from the registry 25.However, as the parties get to know one another, the trust level between them will increase and their reliance on the registry 25 will decrease.\",\n \"Further Embodiments The first embodiment discussed above made no use of the registry 25 for validation or for payment, as it was assumed that only a low level of validation was required and payment was made directly from the purchaser smart-card 3 to the vender smart-card 23.However, either or both parties to a transaction may wish to validate the other party with the registry 25 or make payment to or from a registry account card 33.Where the registry 25 is to be involved in a transaction, session IDs are generated as described above with reference to FIG.\",\n \"7, to include the registry (for validation data) and each registry account card used (for payment) in a secure session.\",\n \"Where the registry 25 is to interact with the purchaser and vendor, a separate secure session with its own session ID pair is generated between the registry 25 and the vendor smart-card 23 and between the registry 25 and the purchaser smart-card 3.However, if only one of the vendor and purchaser needs to contact the registry 25 then a session having a new session ID pair will be generated between the registry 25 and the vendor smart-card 23 or the purchaser smart-card 3.Thus the overall transaction will be made up of a number of sessions, each session having its own session ID pair.\",\n \"In this embodiment, to ensure a common reference between the various sessions making up the transaction, each party uses the same session ID in each session of a given transaction.\",\n \"For example, if the purchaser smart-card 3 has already established a session with the vendor smart-card 23 (and has thus created a purchaser session ID), and then requires a session with the registry 25, the purchaser session ID used in the session with the registry 25 will be the same as the purchaser session ID already used in the session with the vendor smart-card 23.Obviously, the vendor session ID and the registry session ID will be different to one another.\",\n \"Referring now to FIG.\",\n \"12, there will be described a method of performing validation of the purchaser using validation data held by the registry 25 in which the purchaser is informed of the decision to validate at the registry 25 and has the option to refuse such validation.\",\n \"Such processing would take the place of the processing described with reference to FIG.\",\n \"8 above in the first embodiment.\",\n \"It is assumed in the method shown in FIG.\",\n \"12 that the registry 25 and the vendor smart-card 23 have already established a secure session by means of a process similar to that set out above with reference to FIG.\",\n \"7.At step S11-1, the level of validation required is determined as in step S7-1.Next, at step S11-3, the vendor terminal 21 transmits a request for validation at the registry 25 to the purchaser terminal 5.At step S11-5, the purchaser terminal 5 receives the request and at step S11-7 decides whether or not to agree to the level of validation requested.\",\n \"This decision is either made automatically by the purchaser terminal 5 according to predetermined rules set by the purchaser or manually by presenting the request to the purchaser on monitor 9 and awaiting acceptance or refusal of the request from the purchaser via the keyboard 11 or mouse 9.If the requested level of validation is not acceptable, then processing continues at step S11-9 where the purchaser terminal 5 transmits a disapproval of the validation request to the vendor terminal 21.The disapproval is received by the vendor terminal 21 at step S11-11, following which the transaction process terminates.\",\n \"On the other hand, if the requested level of validation is agreed, then processing continues at step S11-13, at which step the purchaser terminal 5 transmits an approval of the validation request to the vendor terminal 21.Following receipt of the validation approval at step S11-15, the vendor terminal 21 transmits a validation request (identifying the purchaser) to the registry 25 at step S11-17.At step S11-19, the registry 25 receives the validation request from the vendor terminal 21 and at step S11-21 transmits the appropriate validation data back to the vendor terminal 21.The validation data is received by the vendor terminal 21 at step S11-23.Processing then proceeds as described above with reference to FIG.\",\n \"8 such that the processing of steps S11-25 to S11-33 corresponds to the processing of steps S7-11 to S7-19.Another alternative to the processing of FIG.\",\n \"8 is shown in FIG.\",\n \"13.In this Figure, validation of the purchaser is performed at the registry 25 but the purchaser terminal 5 is given no notice that the vendor terminal 21 is to receive validation data from the registry 25.It is assumed in the following description that the registry 25 and the vendor smart-card 23 have already established a secure session by means of a process similar to the one described with reference to FIG.\",\n \"7 above.\",\n \"At step S12-1, the level of validation required is determined as in step S7-1.At step S12-3, the vendor terminal 21 transmits to the registry 25 a request for validation of the purchaser, which request is received by the registry 25 at step S12-5.At step S12-7, the registry 25 transmits the validation data, which is then received by the vendor terminal 21 at step S12-9.Processing then continues as for FIG.\",\n \"8 with the processing of steps S12-11 to S12-19 corresponding to the processing of steps 7-11 to steps 7-19.A further method of validation will now be described with reference to FIG.\",\n \"14.In this Figure it is the vendor which is being validated, however it should be understood that all methods of validation can be used to validate either the purchaser or the vendor or both.\",\n \"It is assumed in the following description that the registry 25 and the vendor smart-card 23 have established a secure session and that the registry 25 and the purchaser smart-card 3 have established a secure session by means of processing similar to that described with reference to FIG.\",\n \"7 above.\",\n \"At step S13-1, the level of validation required is determined as for step S8-1.At step S13-3, the purchaser terminal 5 transmits to the registry 25 a request for validation of the vendor.\",\n \"Following receipt of the validation request by the registry 25 at step S13-5, the registry 25 transmits to the vendor terminal 21 at step S13-7 a request for permission to validate.\",\n \"The request is received at step S13-9 by the vendor terminal 21, and at step S13-11 it is decided whether to agree to the level of validation requested in substantially the same manner as described above with reference to FIG.\",\n \"12.If the level of validation is not agreed to then processing continues at step S13-13 where the vendor terminal 21 transmits a disapproval of the validation request to the registry 25, following which the registry 25 receives the validation disapproval at step S13-15.The transaction processing then ends.\",\n \"On the other hand, if it is decided at step S13-11 that the level of validation requested is agreed, then at step S13-17, the vendor terminal 21 transmits a validation approval to the registry 25, which approval is received by the registry 25 at step S13-19.At step S13-21, the registry 25 then transmits the validation data to the purchaser terminal 5 which receives the validation data at step S13-23.Following this, the processing continues as in FIG.\",\n \"9 with the processing of steps S13-25 to S13-33 corresponding to the processing of step S8-11 to S8-19.It is also possible for payment to take place in a number of different ways.\",\n \"For example, as mentioned above, the registry 25 may hold account cards 33p, 33v on behalf of users of the payment system in a smart-card server 27.The presence of these account cards make it possible for a prospective purchaser to have a large volume of funds available to spend using the payment system without having to have those funds tied into the card which they carry, which card could be lost or stolen thereby denying the user access to those funds.\",\n \"It is therefore possible that a purchaser may wish to make a payment from his or her account card 33p or a vendor may wish to have a payment made to his or her account card 33v.\",\n \"If such a payment is to take place, then this will be agreed between the parties before payment is made.\",\n \"If the purchaser or the vendor decides to make payment from or to receive payment from a registry smart-card 33, then the purchaser or the vendor smart-card is informed of this and in response, it provides account details of the appropriate registry smart-card 33, from its registry data store 82.This information is then transmitted to the other smart-card so that the appropriate secure session can be established between the other smart-card and the registry smart-card.\",\n \"Examples of these will now be described with reference to FIGS.\",\n \"14 to 16.Referring now to FIG.\",\n \"15, there will be described payment and receipting steps to replace those described above with reference to FIG.\",\n \"11 where a purchaser pays from funds held on their smart-card 3 to a registry account smart-card 33v held on the vendors behalf.\",\n \"It is assumed in the following description that the vendor registry account card 33v and the vendor smart-card 23 have already established a secure session and that the vendor registry account card 33v and the purchaser smart-card 3 have already established a secure session by means of processing similar to that described with reference to FIG.\",\n \"7 above.\",\n \"At step S14-1, the vendor smart-card 23 transmits to the purchaser smart-card 3 a request for payment, which request is received at step S14-3.At step S14-5, the purchaser smart-card 3 transmits the appropriate payment to the vendor account card 33v at the vendor account card 33v registry 25, which payment is received by the vendor account card 33v at step S14-7.At step S14-9, the vendor account card 33v then transmits to the vendor smart-card 23 an acknowledgement that the payment has been made which acknowledgement is received at step S14-11 by the vendor smart-card 23.The receipting process now continues as in FIG.\",\n \"11 with the processing of steps S14-13 to S14-25 corresponding to the processing of steps S10-9 to S10-21.Referring now to FIG.\",\n \"16, there will be described a method of payment to the vendor's smart-card 23 from the purchaser's registry account card 33p.\",\n \"It is assumed in the following description that the purchaser smart-card 3 has already transmitted account details of the purchaser's registry account card 33p to the vendor smart-card 23, that the registry account card 33p and the vendor smart-card 23 have established a secure session and that the purchaser registry account card 33p and the purchaser smart-card 3 have already established a secure session by means of processing similar to that described with reference to FIG.\",\n \"7 above.\",\n \"At step S15-1, the vendor smart-card 23 requests payment from the purchaser smart-card 3.Following receipt of the request for payment by the purchaser smart-card 3 at step S15-3, the purchaser smart-card 3 transmits, at step S15-5, to the purchaser's registry account card 33p, an instruction to make the payment.\",\n \"At step S15-7, the registry account card 33p receives the payment instruction and at step S15-9, it transmits the payment to the vendor smart-card 23.The payment is received by the vendor smart-card 23 at step S15-11, following which the receipting procedure proceeds as in FIG.\",\n \"11, whereby the processing of steps S15-13 to steps S15-25 corresponds to the processing of steps S10-9 to S10-21.Referring now to FIG.\",\n \"17, there will be described the processing where payment is made from the purchaser's registry account card 33p to the vendor's registry account card 33v.\",\n \"It is assumed in the following description that the purchaser smart-card 3 and the vendor smart-card 23 have received details of the other's registry account cards 33p,33v; that the two registry account cards 33p and 33v have already established a secure session between each other; that the purchaser registry account card 33p and the purchaser smart-card 3 have already established a secure session between each other; and that the vendor registry account card 33v and the vendor smart-card 23 have already established a secure session between each other.\",\n \"At step S16-1, the vendor smart-card 23 transmits a request for payment to the purchaser smart-card 3, which request is received at step S16-3.At step S16-5, the purchaser smart-card 3 transmits a payment instruction to the purchaser's registry account card 33p, which payment instruction is received at step S16-7.The purchaser account card 33p then effects the payment to vendor's registry account card 33v at step S16-9.At step S16-11, the vendor's registry account card 33v transmits a payment acknowledgement to the vendor smart-card 23 which acknowledgement is received at step S16-13.Thereafter, the receipting process between the purchaser smart-card 3 and the vendor smart-card 23 continues as in FIG.\",\n \"11, with the processing of steps S16-15 to S16-27 corresponding to the processing of steps S10-9 to S10-21.A further optional feature of the payment system described in any of the above embodiments is that a smart-card for use with the payment system may hold funds in a number of different currencies.\",\n \"It will be possible to load funds onto a purchaser's smart-card 3 or registry account card 33p in the required currency or it would be possible to convert funds held on the card.\",\n \"Conversion between currencies could take place on a particular user card 3 or 23 or, as would be preferable from a fraud control point of view, conversion could be undertaken on a user's registry account card 33 under the control of the registry 25 to prevent fraudulent currency creation.\",\n \"The process of currency conversation does not entail the actual conversion of one currency into another, rather it implies debiting an amount in one currency and crediting the equivalent amount in another currency.\",\n \"The implications of this are twofold; 1) external (off card) funds must be held in the currency to be supplied; 2) the exchange rate will be externally (off card) supplied.\",\n \"An example of how conversion of funds within a registry account card 33 could be accomplished by a user will now be described with reference to FIG.\",\n \"18.Before the currency conversion process may commence, a secure session between the user's own card 3 or 23 and the registry card 33p or 33v, respectively must be established.\",\n \"This is achieved in the same manner as described above with reference to FIG.\",\n \"6.Thus steps S17-1 to S17-19 correspond to steps S6-9 to S6-27 in that the user card 3 transmits its public key to the registry card which, following receipt of the user card public key, generates a registry session ID in the same manner as the session IDs were generated with reference to FIG.\",\n \"7 above.\",\n \"The registry smart-card 33p or 33v then encrypts the registry session ID with the cardholder public key and transmits the encrypted session ID and the registry public key back to the user smart-card 3.The user smart-card 3 then generates a user session ID, encrypts it with the registry public key and transmits the encrypted user session ID to the registry.\",\n \"Once the registry smart-card 33p or 33v has received the encrypted user session ID, it transmits a message to the user card 3 stating that the session ID was received successfully.\",\n \"Following this, at step S17-21 the cardholder terminal 5 transmits a request for conversion of account funds from a first currency to a second currency which request is received by the registry 25 at step S17-23.At step S17-25, the registry 25 performs a check to determine whether the account card 33 holds sufficient funds to perform the currency conversion.\",\n \"If it is determined that sufficient funds are not present, then processing continues at step S17-27, where the registry 25 transmits to the cardholder terminal 5 that there are insufficient funds in the account card 33p to perform a conversion.\",\n \"This is received by the cardholder terminal 5 at step S17-29, following which the conversion procedure terminates.\",\n \"On the other hand, if it is determined at step S17-25 that there are sufficient funds in the account to perform the currency conversion then at step S17-31 the funds are converted and at step S17-33 the registry 25 transmits to the cardholder terminal 5 a confirmation that the funds have been converted and a note of the funds totals in the relevant currencies available on the registry account card following the conversion.\",\n \"This is received by the card holder terminal 5 at step S17-35, following which the currency conversion procedure terminates.\",\n \"A further feature which may be applied to the payment system according to any of the described embodiments is that transaction logging may be undertaken.\",\n \"At the end of a given transaction, both the vendor and purchaser will have stored in the transaction log store 80 of their respective smart-cards 3, 23 a record of all of the data transferred during the transaction.\",\n \"This record will include the goods or services specified, any validation data concerning the purchaser or vendor, any delivery data and payment data.\",\n \"Each party to the transaction therefore has a complete record of that transaction.\",\n \"A limited number of such transaction records may be held by the smart-card 3, 23 to provide a short-term transaction log.\",\n \"However, due to the memory limitations of smart-cards, it is not likely to be possible for the smart-card to store a complete log of all transactions in which the particular smart-card 3, 23 is involved.\",\n \"Therefore, the smart-cards are programmed to allow the transaction records to be copied from the transaction log store 80 into a full transaction log within a personal computer or the like.\",\n \"For example, a vendor may set up an automatic process through their usual vending terminal such that at the completion of any transaction, a copy of that transaction record is made on a central transaction log held within the vendor terminal or in a further terminal which is connected to the vendor terminal.\",\n \"A purchaser may for example, have a home computer comprising a smart-card reader into which the user smart-card 3 may be inserted such that records of purchases made using the smart-card 3 may be transferred to a log on the home computer.\",\n \"In addition, the registry 25 may be configured to automatically save details of all transactions in which it is involved in order to provide a centralised accountability for the payment system.\",\n \"In the above embodiments, different levels of validation were described.\",\n \"Some practical trading examples will now be described that illustrate how these different levels of validation would be used.\",\n \"VALIDATION LEVEL EXAMPLES Internet Trading Example 1 In this example, a user of the payment system wishes to purchase weather forecast data from the UK Meteorological Office.\",\n \"The purchaser knows he is at the Meteorological Office website and trusts them to deliver the requested information.\",\n \"There is no reason for the purchaser to carry out any further checking on the vendor.\",\n \"The Meteorological Office does not care who the purchaser is or where they are located.\",\n \"The information to be purchased is not restricted in any way.\",\n \"In addition, the Meteorological Office will collect payment from the purchasers smart-card before shipping the requested information.\",\n \"So the vendor requires no further information about the purchaser and an immediate anonymous trade can take place subject to both parties indicating their satisfaction with the price and terms of trade.\",\n \"Internet Trading Example 2 In this example, the purchaser wishes to buy videos from an internet discount store.\",\n \"In this case the value of the trade will be low but checking is required by both parties.\",\n \"The vendor will wish to assure itself that the purchaser is of a suitable age to purchase the videos requested, while the purchaser will wish to validate the credentials and location of the vendor to ensure that the vendor will not simply take the purchaser's money and disappear, never delivering the goods.\",\n \"The vendor could simply ask a purchaser for their age and address but there would be no support for this information, which therefore could not be trusted.\",\n \"The vendor would therefore check the credentials of the purchaser with the registry 25 and if the purchaser was not registered or refused to permit the information to be released, the vendor could refuse to sell the products to this purchaser.\",\n \"If the information were provided, only a low confidence level would probably be required and the registry 25 would provide no warranties or guarantees.\",\n \"The purchaser would wish to be assured that the vendor can be trusted.\",\n \"To achieve this, the purchaser would check that the vendor had a registry account and that the vendor would allow the registry 25 to release/confirm the address of the vendor.\",\n \"Only under these circumstances would the purchaser agree to make the trade.\",\n \"When both parties are satisfied with the information supplied, the sale price, the transaction charges and the terms of trading, the sale would be completed and value transferred.\",\n \"In subsequent trades between the two parties, it is likely that the purchaser will have built up a level of trust in the vendor such that the purchaser will not seek any validation from the registry.\",\n \"Internet Trading Example 3 This example concerns the purchase of fine art.\",\n \"In this circumstance a great deal of checking and validation will be required.\",\n \"Even though the vendor will be paid immediately, the vendor may need to verify the nationality and domicile of a purchaser.\",\n \"The vendor may also wish to avoid being part of a possible money laundering exercise and will therefore seek the most exhaustive credentials concerning a purchaser with the highest level of confidence.\",\n \"The purchaser will also need to have more guarantees than simply the credentials of the vendor.\",\n \"Since the fine art cannot be authenticated until it is shipped (or the benefit of using the internet will be lost), and since the goods may be damaged in transit, the purchaser may seek a cash back guarantee from the smart-card registry 25.For this purpose, the registry 25 may be underwritten to supply such a guarantee with the properly authenticated vendors but of course is likely to levy a high transaction charge for this service.\",\n \"Once all parties have agreed to the levels of confidence they have been given, agreed prices, transaction charges and terms and conditions, the trade may be completed.\",\n \"Given the high value of the deal, it is likely that payment will be made from an account card held by the registry on the purchaser's behalf, rather than his or her personal card.\",\n \"In addition, it may be possible for the registry operator to function as a credit providing service such that users of the payment system may be able to borrow money from the registry to be paid back at a later date as with a traditional credit card 25.Real World Trading Example 1 For the purpose of purchasing goods such as a newspaper the smart-card of the payment system according to the present invention may be used as a “traditional” electronic purse at any shop accepting the card.\",\n \"To make a newspaper purchase the cardholder will simply use his or her card in an anonymous manner to make the purchase using a purchaser smart-card reader provided within the shop.\",\n \"Real World Trading Example 2 For the purchase of a larger value item such as a television, the purchaser may have sufficient value on their personal card to make payment or they may indicate their desire to pay from the registry account card in much the same way as a traditional bank debit card payment.\",\n \"In either event, the vendor will wish to validate the credentials of the purchaser to check that the card has not been lost or stolen and that a registry account is not being fraudulently used.\",\n \"In the case of account payment, the vendor will also take payment from the registry 25.For this purpose the vendor will need a simple smart-card terminal capable of connectivity either directly to the registry or via the internet.\",\n \"Such terminal may be similar to the bank/credit card terminal currently in use for debit and credit card payments.\",\n \"Since the purchaser is physically in the vendor's shop it is unlikely that they will seek any further accreditation of the vendor, however this is always possible.\",\n \"For example, the purchaser may wish to obtain guarantees about the delivery date and goods quality for purpose made goods.\",\n \"To achieve this, the purchaser will make use of a vendor's terminal to contact the registry and seek the necessary guarantees.\",\n \"Once all terms have been agreed, the purchaser can authorise payment to be made in exactly the same way as in the case with internet trading.\",\n \"Real World Trading Example 3 Booking or purchasing a ticket from an unattended terminal is the same as internet trading.\",\n \"For ticket purchases, it will be possible for a physical paper ticket to be issued via the terminal, for a ticket to be delivered to the vendor following the purchase transaction at the terminal or for an electronic ticket to be written to the purchaser's smart-card.\",\n \"This is possible because a smart-card is not restricted to holding programs and/or data for a single purpose but may have a number of separate application entities on the same card.\",\n \"In particular, technologies such as Sun Microsystems Inc's JavaCard™ technology allow applications to be written to a smart-card at any time.\",\n \"Thus the user of a smart-card equipped with programming to perform processing to enable it to participate in the payment system of the present invention could decide that the ability to have electronic tickets written to his or her card would be useful and therefore arrange for software to enable that function to be written to his or her card at any time.\",\n \"Although it has been described above with reference to FIGS.\",\n \"1 to 16 that the purchaser terminal 5 and the vendor terminal 21 are physically separated and connected via the internet, it is possible that a cardholder may wish to use their smart-card at the vendor's location.\",\n \"This situation is discussed in the above trading examples, and is shown in a simplified form in FIG.\",\n \"19.In FIG.\",\n \"19, the vendor terminal 21 having a monitor 9, a keyboard 11 and a mouse 13, which terminal may be combined with or a part of a shop cash register or till, is equipped with a smart-card reader 31 into which the vendor smart-card 23 may be inserted.\",\n \"For security purposes it may be preferable for the smart-card slot 31 to be physically separated from the terminal 21 and connected via a cable or other data communications link because the terminal 21 may be in an area accessible to customers and having the smart-card 23 in such a location could be perceived to be a security risk.\",\n \"As before, the vendor terminal 21 is equipped with a modem 15 by means of which it may communicate with the registry terminal 25 by transmitting signals 17 via the internet 19 or by a telephone line.\",\n \"Also connected to the vendor terminal 21 is a user smart-card reader 34 into which the user smart-card 3 may be inserted by a purchaser when a purchase is to be made using the payment system of the present embodiment.\",\n \"In such circumstances, there is obviously no need for communication between the purchaser smart-card 3 and the vendor smart-card 23 to take place via the internet and thus the same process as described above with reference to flow diagrams 4 to 16 may be performed as shown but with communication between the cards 3 and 23 taking place via the cables connecting the user smart-card reader 34 to the vendor smart-card reader 31.It should be noted that in the above-described arrangement, the vendor and purchaser smart-cards still create a secure session to wrap the transaction to provide full transaction accountability and traceability, to provide a secure channel between the cards to transfer funds, and to enable data to be certified, all as described above.\",\n \"In the event that either the purchaser or the vendor should wish to validate the other party at the registry or make payment from or receive payment to a registry account card this may be achieved using the internet connection to the registry terminal 25.Referring to FIG.\",\n \"20, there is shown a block diagram of the main functional components of the vendor terminal 21 used in this embodiment.\",\n \"As before, a CPU making use of a working memory 43 to perform instructions stored in software store 47, a keyboard and mouse interface 49 to receive input from a user and a monitor interface 51 to output information to a user are present and are connected via a data bus 45.Also connected via the data bus 45 is the modem 55 for communication with the registry terminal 25 via the internet 19 or telephone line.\",\n \"The vendor smart-card reader interface 59 and the purchaser smart-card reader 61 are also connected to the data bus 45.Although it has been described above that the various terminals communicate with one another via the internet, the terminals may be connected together via a Local Area Network (LAN) or a Wide Area Network (WAN).\",\n \"In addition, it is possible that the terminals could perform a so-called “tunnelling” operation to create a Virtual Private Network (VPN) between the terminals across the internet, or a direct dial telephone link between the terminals may be used.\",\n \"Although it has been described above that where particular validation data or particular delivery terms are decided to be unacceptable the transaction process terminates, further negotiations between the two parties could be undertaken to resolve such situations.\",\n \"For example, a prospective purchaser may select a product and request next day delivery, but then decide that the cost for next day delivery is more than he or she is willing to pay.\",\n \"It could be made possible for the prospective purchaser to request to be offered a cheaper delivery service which they might then accept.\",\n \"Although it has been described above, particularly with reference to FIG.\",\n \"5, that validation of a purchaser is performed before validation of the vendor, it is not essential that the validations are performed in this order and therefore validation of a vendor could be performed before validation of the purchaser or even at the same time.\",\n \"Although, it has been described above with reference to FIG.\",\n \"5 that a step is taken to determine whether or not to validate the purchaser or vendor followed by which the level of validation required is determined, it is possible that these two steps could be combined into one single operation, wherein if no validation is required the steps to achieve validation could be omitted.\",\n \"Although it is described above with reference to FIG.\",\n \"9 that a purchaser is required to enter delivery data when required, it is possible that such data could be stored within the purchaser terminal or purchaser smart-card and the transfer of such data to the purchaser terminal could be achieved without reference to the purchaser.\",\n \"Although it is described above that the registry 25 stores validation data, it is possible that the validation data could be stored on the registry account card 33v, 33p.\",\n \"Although it is described above that a single registry 25 is present, it is possible that a number of registries could exist.\",\n \"In this case it would become necessary to establish which registry should be queried to receive validation data from the party to be validated.\",\n \"For simplicity, this information is preferably stored on the party's smart-card.\",\n \"Although it has been described with reference to FIG.\",\n \"4 that delivery data is transmitted after validation, it is entirely possible to include delivery data within the validation data.\",\n \"This is a favourable modification to the above system as it is likely that validation data would include address data and therefore duplication of data can be reduced and thus transaction processing time can be reduced.\",\n \"The delivery data would always be transmitted after establishing the session, but need not necessarily be encrypted.\",\n \"Although it has been described above with reference to FIG.\",\n \"7 that a specific security arrangement is used to establish the secure session, it is to be noted that the security algorithms used by the smart-cards and the operation of the security engine on the smart-cards does not matter, provided that an asymmetric key system of the type used in public key encryption is used.\",\n \"Examples of encryption schemes which could be used are RSA type algorithms and elliptic curve techniques.\",\n \"However, it is not an open public key system requiring a certification authority to validate ownership, rather it is a private system known only to the smart-cards themselves.\",\n \"Although it has been described above with reference to FIG.\",\n \"7 that all data passing between the terminals (and the registry) will be encrypted by the smart-card 3, 23, 33 in the terminal (or registry) before transmission, it is not necessary for all data to be encrypted.\",\n \"Only the data which the parties do not wish to be general public knowledge (which could happen if the data were intercepted) and the data which requires certification as to its origin and validity (such as validation data), and therefore requires a “digital signature”, need be encrypted.\",\n \"As those skilled in the art will appreciate, the function of the secure session IDs of digitally signing the transmitted data to certify validity and origin could be achieved by methods other than that described above.\",\n \"In particular, a number of techniques of so-called “digital water-marking” exist, which could be used to certify the data, or encryption keys themselves could be used as signatures.\",\n \"Although it has been described above that the various terminals have specific components and arrangements, it is sufficient for the performance of the embodiments that any stand alone terminal has a card reader, a means for presenting data to and requesting data from a user, means for receiving user input and means for communicating with other terminals.\",\n \"Thus it is not necessary for the terminal to be a desk-top computer.\",\n \"It is possible that terminals may take the form of railway, airline or bus ticket vending machines, public information booths or terminals, theatre or cinema ticketing booths, shop tills, portable computers, hand-held computers and information appliances, telephones and tourist attraction entry gates, for example.\",\n \"As those skilled in the art will appreciate, the present invention can be used not only from a purchaser's own home terminal but from terminals in so-called internet cafes and in other public terminals.\",\n \"Although it has been described above with reference to FIGS.\",\n \"11 to 16 that the registry or a registry account card is included in the secure session before a request for validation data or payment is made to the registry or account card, it is possible that the registry or registry account card will only be included in the secure session when validation data or payment is requested.\",\n \"Although it has been described above that a user would hold funds on their user smart-card and may hold further funds on their registry account card, it is possible to operate the entire system in the manner of a credit card system whereby a user holds no funds on their own smart-card or on their registry account card, but every time they wish to make a purchase requesting payment from the registry account, and funds being provided to the registry account by the registry on a credit basis with the money supplied as credit to be repaid by the user according to predetermined practices.\",\n \"It is also possible for the system to be operated in a combination of a stored cash and credit based system whereby a user may store small amounts of cash on their personal smart-card or on their registry account card, but for large purchases make use of a credit arrangement with the registry provider.\",\n \"Such a system would address the problem of previous smart-card systems whereby funds held on the card are “locked in” to the card.\",\n \"The smart-cards used to implement the present invention may use any standard interface.\",\n \"There is an International Standard (ISO7816-3) which governs the so-called contact interface where the microchip on the smart-card interfaces with a reader by means of physical contacts between the microchip on the smart-card and the reader.\",\n \"There is a further International Standard (ISO14443) which governs the so-called contactless interface.\",\n \"In this interface, the microchip on the smart-card and the reader have no physical connection, all communication between the reader and the smart-card, including supply of power to the smart-card, is performed using radio transmissions.\",\n \"It is also possible to implement the invention using a smart-card capable of using both of the above interface types.\",\n \"As described above, the smart-cards 3,23,33 may be implemented using multi-programmable technologies such as Sun Microsystems Inc's JavaCard™ technology.\",\n \"It is therefore possible for a person or organisation already possessing a smart-card conforming to such a multi-programmable technology to have software to enable the smart-card as a smart-card 3,23,33 according to any of the above embodiments loaded onto their smart-card.\",\n \"Such software loading could be performed by an authority such as a registry operator, or the software could be transmitted to the smart-card owner via the internet or similar connection, or on a storage medium such as a floppy disk or CD-ROM for the smart-card owner to use to load the software onto the smart-card themselves.\",\n \"Although it has been described above that the session IDs should be generated using a time stamp, a random number, a terminal ID and a smart-card ID, it is not necessary that the session ID be generated in this manner for the successful operation of the embodiments.\",\n \"As one skilled in the art would appreciate, any method of generating a new, unique session ID for each transaction that a particular smart-card 3,23,33 is involved in, could be used.\",\n \"This method could involve using less than all of the above parts of the session ID described in the above embodiment, or other values such as account holder identifiers or a transaction counter could be used.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10474050"}}},{"rowIdx":782,"cells":{"text":{"kind":"list like","value":[["Filter cartridge for a filter assembly","A filter assembly including a replaceable filter cartridge for use with a reusable filter canister.","The filter cartridge includes a cylindrical filter media having top and bottom ends respectively mounted within top and bottom end caps.","The bottom end cap includes a sidewall that surrounds at least a lower portion of the filter media.","A contaminant containment chamber is defined between the sidewall and an outer side of the filter media.","When the cartridge is mounted within the canister, the radial sealing element engages an inner surface of the canister to provide a circumferential seal.","The sidewall also includes a living hinge positioned below the sealing element."],["1.A filter cartridge comprising: a filter media having a top end and a bottom end; a top end cap mounted at the top end of the filter media; a bottom end cap mounted at the bottom end of the filter media, the bottom end cap including a bottom wall and a side wall, the side wall extending upwardly from the bottom wall and surrounding at least a lower portion of the filter element, the side wall having a top end; a circumferential sealing element that projects radially outwardly from the side wall at a location adjacent the top end of the side wall; and a circumferential living hinge defined by the sidewall at a location below the circumferential sealing element, wherein the living hinge is formed by a ring of reduced cross sectional thickness provided at the sidewall.","2.The filter cartridge of claim 1, wherein the sidewall is spaced from an outer surface of the filter media such that a contaminant containment chamber is defined between the sidewall and the filter media.","3.The filter cartridge of claim 2, wherein the bottom wall defines a plurality drain openings located beneath the contaminant containment chamber.","4.The filter cartridge of claim 3, further comprising an outer filter element for filtering liquid that drains from the contaminant containment chamber through the drain holes.","5.The filter cartridge of claim 4, wherein the outer filter element is annular and is mounted at a bottom of the contaminant containment chamber.","6.The filter cartridge of claim 1, wherein the living hinge is defined by a circumferential notch defined within the sidewall.","7.The filter cartridge of claim 6, wherein the notch has a generally semi-circular cross-section.","8.The filter cartridge of claim 6, wherein the notch is defined in an inner surface of the sidewall.","9.The filter cartridge of claim 1, further comprising at least one relief notch defined through the sealing element.","10.The filter cartridge of claim 9, further comprising a plurality of relief notches defined through the sealing element.","11.The filter cartridge of claim 10, wherein the relief notches are uniformly spaced about the circumference of the sealing element.","12.The filter element of claim 11, wherein the relief notches include two relief notches spaced apart approximately 180 degrees about the circumference of the sealing element."],["<SOH> BACKGROUND OF THE INVENTION <EOH>Although applicable to a number of fields, the present invention relates primarily to liquid filters such as hydraulic oil filters or engine oil filters.","Filters have long been used to remove contaminants from oil.","It is well known that, with time, oil filters become plugged or clogged with contaminants and are required to be replaced.","A typical oil filter assembly includes a housing (e.g., a bowl or canister) containing a cylindrical filter element.","The cylindrical filter element typically defines a central longitudinal opening in which a central core is mounted.","The core provides support for preventing the filter element from collapsing inward during use.","In use, oil is directed from a system into an annular channel defined between the housing and the outer surface of the filter element.","Pressure from the system forces the oil from the annular flow passage, through the filter element, and into the central opening of the filter element.","From the central opening of the filter element, the filtered oil is discharged from the filter assembly.","Disposable or “throw-away” type fluid filters are well known in the art.","In this type of filter assembly, the filter element is permanently sealed within the housing.","When the filter element becomes clogged with contaminants, the entire filter assembly is discarded and replaced with a new unit.","Oil filter assemblies having replaceable filter elements are also known in the art.","Filter assemblies with replaceable filter elements are desirable because the housing can be reused thereby reducing the amount of waste that enters the waste stream.","Consequently, as landfill costs increase, filter assemblies having replaceable filter elements will become increasingly more desirable.","Contamination is an important consideration relating to filter assemblies having replaceable filter elements.","For example, prior art systems having replaceable filter elements can experience the accumulation of contaminants within the reusable housings that cover the replaceable filter elements.","If this occurs, the “clean” side of new filter elements inserted within the contaminated housings can immediately become contaminated.","There is a need in the art to prevent this type of contamination from occurring."],["<SOH> SUMMARY OF THE INVENTION <EOH>One aspect of the present invention relates to a filter assembly including a replaceable filter cartridge for use with a reusable filter canister.","The filter cartridge includes a cylindrical filter media having top and bottom ends respectively mounted within top and bottom end caps.","The bottom end cap includes a sidewall that surrounds at least a lower portion of the filter media.","A contaminant containment chamber is defined between the sidewall and an outer side of the filter media.","A radial sealing element is located adjacent a top end of the sidewall.","When the cartridge is mounted within the canister, the radial sealing element engages an inner surface of the canister to provide a circumferential seal.","The sidewall also includes a living hinge positioned below the sealing element.","In use of the filter assembly, contaminants are accumulated on the outer side of the filter element.","Larger contaminants fall downwardly and are captured in the contaminant containment chamber defined by the lower end cap.","The radial sealing element prevents contaminants from by-passing the contaminant containment chamber.","When the filter element is fully loaded with contaminants, the filter cartridge can be removed from the canister and replaced.","The living hinge facilitates removal of the filter cartridge from the canister by reducing friction between the radial sealing element and the inner surface of the canister.","A variety of advantages of the invention will be set forth in part in the description that follows, and in part will be apparent from the description, or may be learned by practicing the invention.","It is to be understood that both the foregoing general description and the following detailed description are illustrative and explanatory only and are not restrictive of the invention as claimed."],["This application is being filed as a PCT international patent application in the name of Donaldson Company, Inc., a U.S. corporation (applicant for all designations except the U.S.), and in the names of the Stan M. Koehler, Brent A. Gulsvig, and John R. Hacker, all U.S. citizens and residents (applicants for the U.S. designation only), on Apr.","2, 2002, designating all countries.","FIELD OF THE INVENTION The present invention relates generally to filters.","More particularly, the present invention relates to liquid filters (e.g., hydraulic fluid filters, engine oil filters or fuel filters) having replaceable filter elements (i.e., filter cartridges).","BACKGROUND OF THE INVENTION Although applicable to a number of fields, the present invention relates primarily to liquid filters such as hydraulic oil filters or engine oil filters.","Filters have long been used to remove contaminants from oil.","It is well known that, with time, oil filters become plugged or clogged with contaminants and are required to be replaced.","A typical oil filter assembly includes a housing (e.g., a bowl or canister) containing a cylindrical filter element.","The cylindrical filter element typically defines a central longitudinal opening in which a central core is mounted.","The core provides support for preventing the filter element from collapsing inward during use.","In use, oil is directed from a system into an annular channel defined between the housing and the outer surface of the filter element.","Pressure from the system forces the oil from the annular flow passage, through the filter element, and into the central opening of the filter element.","From the central opening of the filter element, the filtered oil is discharged from the filter assembly.","Disposable or “throw-away” type fluid filters are well known in the art.","In this type of filter assembly, the filter element is permanently sealed within the housing.","When the filter element becomes clogged with contaminants, the entire filter assembly is discarded and replaced with a new unit.","Oil filter assemblies having replaceable filter elements are also known in the art.","Filter assemblies with replaceable filter elements are desirable because the housing can be reused thereby reducing the amount of waste that enters the waste stream.","Consequently, as landfill costs increase, filter assemblies having replaceable filter elements will become increasingly more desirable.","Contamination is an important consideration relating to filter assemblies having replaceable filter elements.","For example, prior art systems having replaceable filter elements can experience the accumulation of contaminants within the reusable housings that cover the replaceable filter elements.","If this occurs, the “clean” side of new filter elements inserted within the contaminated housings can immediately become contaminated.","There is a need in the art to prevent this type of contamination from occurring.","SUMMARY OF THE INVENTION One aspect of the present invention relates to a filter assembly including a replaceable filter cartridge for use with a reusable filter canister.","The filter cartridge includes a cylindrical filter media having top and bottom ends respectively mounted within top and bottom end caps.","The bottom end cap includes a sidewall that surrounds at least a lower portion of the filter media.","A contaminant containment chamber is defined between the sidewall and an outer side of the filter media.","A radial sealing element is located adjacent a top end of the sidewall.","When the cartridge is mounted within the canister, the radial sealing element engages an inner surface of the canister to provide a circumferential seal.","The sidewall also includes a living hinge positioned below the sealing element.","In use of the filter assembly, contaminants are accumulated on the outer side of the filter element.","Larger contaminants fall downwardly and are captured in the contaminant containment chamber defined by the lower end cap.","The radial sealing element prevents contaminants from by-passing the contaminant containment chamber.","When the filter element is fully loaded with contaminants, the filter cartridge can be removed from the canister and replaced.","The living hinge facilitates removal of the filter cartridge from the canister by reducing friction between the radial sealing element and the inner surface of the canister.","A variety of advantages of the invention will be set forth in part in the description that follows, and in part will be apparent from the description, or may be learned by practicing the invention.","It is to be understood that both the foregoing general description and the following detailed description are illustrative and explanatory only and are not restrictive of the invention as claimed.","BRIEF DESCRIPTION OF THE DRAWINGS FIG.","1 is an exploded, perspective view of a filter assembly constructed in accordance with the principles of the present invention, the assembly includes a canister and a replaceable filter cartridge; FIG.","2 is an assembled view of the filter assembly of FIG.","1 with the assembly being longitudinally bisected; FIGS.","3A and 3B are exploded, perspective views of the filter cartridge of FIGS.","1 and 2; FIG.","4 is a top view of the filter cartridge of FIGS.","3A and 3B; FIG.","5 is a cross-sectional view taken along sectional line 5-5 of FIG.","4; FIG.","6 is a top, perspective view of the bottom end cap of the filter cartridge of FIGS.","3A and 3B; FIG.","7 is a bottom, perspective view of the bottom end cap of FIG.","6; FIG.","8 is bottom, plan View of the bottom end cap of FIG.","6; FIG.","9 is a cross-sectional view taken along section line 9-9 of FIG.","8; FIG.","9A is a detailed view of FIG.","9 showing a living hinge defined by a sidewall of the bottom end cap; FIG.","10 is a top, perspective view of an alternative bottom end cap that is an embodiment of the present invention; FIG.","11 is a bottom, perspective view of the bottom end cap of FIG.","10; FIG.","12 is a top plan view of the bottom end cap of FIG.","10; FIG.","13 is a cross-sectional view taken along section line 13-13 of FIG.","12; and FIG.","14 is a cross-sectional view taken along section line 14-14 of FIG.","12.DETAILED DESCRIPTION Reference will now be made in detail to preferred embodiments of the present invention that are illustrated in the accompanying drawings.","Wherever possible, the same reference numbers will be used throughout the drawings to refer to the same or like parts.","FIG.","1 shows an embodiment of a filter assembly 20 constructed in accordance with the principles of the present invention.","The filter assembly 20 includes a reusable canister 22 (i.e., a bowl or housing) adapted for connection to a filter head 45 (shown in FIG.","2).","The filter head 45 is part of a pressurized fluid/liquid system such as a fuel system, an engine lubrication system or a hydraulic oil system.","The filter assembly 20 also includes a replaceable filter cartridge 34 that mounts within the canister 22.The filter cartridge 34 is adapted for filtering contaminants from the pressurized fluid/liquid system.","In use, the filter assembly 20 is connected to the filter head 45.Once connected, liquid from the pressurized system is directed from the filter head 45 to an outer side of the filter cartridge 34 (see arrows 200).","From the outer side of the cartridge 34, the liquid flows through the filter cartridge 34 to an inner “clean” side of the filter cartridge 34 (see arrows 202).","Contaminants within the liquid are accumulated on the outer “dirty” side of the filter cartridge 34.Filtered liquid from the “clean” side of the filter cartridge 34 is re-introduced into the pressurized system (see arrow 204).","Referring to FIG.","1, the assembly 20 also includes a perforated core 24.As shown in FIG.","2, the core 24 is preferably fixably mounted within the canister 22.For example, the core 24 can be press fit over a cylindrical mounting post 36 formed at a bottom end 32 of the canister 22 (see FIG.","2).","Adhesive can also be used to better secure the core 24 to the post 36.When assembled, as shown in FIG.","2, the filter cartridge 34 fits over the core 24 such that the core 24 provides radial reinforcement to the filter cartridge 34.The radial reinforcement provided by the core 24 prevents the filter cartridge 34 from collapsing radially inwardly during use.","Referring again to FIG.","1, the filter assembly 20 further includes an annular sealing member 26 (e.g., an O-ring) and a plug 32.The annular sealing member 26 is preferably mounted adjacent a top end 28 of the canister 22 for providing a fluid tight seal with the filter head 45.The drain plug 30 is preferably threaded within a drain plug opening 31 (shown in FIG.","2) defined by the bottom end 32 of the canister 22.The canister 22 is preferably a “spin-on” canister 22.By “spin-on”, it is meant that the canister 22 is connected to the engine head 45 by spinning (i.e., twisting, turning or rotating) the canister 22.For example, as shown in FIG.","2, the canister 22 includes internal threads 38 that interlock with corresponding threads 47 defined by the filter head 45.The annular sealing member 26 of the canister 22 is preferably mounted within an annular recess 40 located directly beneath the threads 38.When the canister 22 is fully threaded on the filter head 45, the annular seal 26 preferably engages a cylindrical sealing surface 49 of the filter head 45 to provide a fluid tight seal.","Still referring to FIG.","2, the filter assembly 20 also includes an inner, annular sealing member 50.The inner, annular sealing member 50 is mounted within an annular recess 52 defined by the filter cartridge 34.When the canister 22 is fully threaded on the filter head 45, a cylindrical extension 54 of the filter head 45 projects partially into the filter cartridge 34.The extension 54 includes a cylindrical sealing surface 55 that engages the annular sealing member 50 to provide a fluid tight seal.","Preferably, the canister 22 has a generally thin-walled construction and is formed from a metal such as aluminum, steel or other suitable material.","The core 24 is also preferably made of a material such as steel or aluminum.","FIGS.","3A and 3B are exploded views of the filter cartridge 34.The filter cartridge 34 includes a cylindrical filter media 60 having top and bottom ends 61 and 62 respectively mounted within top and bottom end caps 64 and 65.A potting material (e.g., a material having adhesive characteristics such as urethane) can be used to seal and secure the top and bottom ends 61 and 62 in their respective end caps 64 and 65.The filter media 60 preferably has a pleated configuration.","In certain embodiments, the filter media 60 can include a pleated paper media adapted for filtering liquids such as fuel, engine oil, hydraulic fluids or other liquid media.","Exemplary types of filter media include sintered metal, cellulose, synthetic media, wire mesh, and glass fibers.","Also, either pleated or unpleated filter media can be used.","Referring to FIG.","3B, the top end cap 64 of the filter cartridge 34 includes an annular channel 66 sized to receive the top end 61 of the filter media 60.The top end cap 64 also preferably includes a handle 68 (best shown in FIGS.","3A and 4) for use in pulling the filter cartridge 34 from the canister 22 during the cartridge removal process.","Further, the top end cap 64 preferably defines the annular recess 52 (shown in FIG.","2) for mounting the annular sealing member 50.Referring to FIGS.","6 and 7, the bottom end cap 65 of the filter cartridge 34 preferably includes a bottom wall 70 and a circumferential sidewall 72 that projects upwardly from the bottom wall 70.The bottom end cap 65 is preferably made of a plastic material such as glass-filled nylon.","Preferably, the bottom end wall 70 and the sidewall 72 are integrally formed as a single unitary piece.","In the depicted embodiment, the sidewall 72 is preferably made of a material that is impervious to liquids (e.g., hydraulic oil).","Referring to FIG.","7, the bottom wall 70 is preferably generally planar.","A plurality of interlock members 74 project outwardly from the bottom side of the bottom wall 70.The interlock members 74 are adapted to engage corresponding interlock members formed in the bottom of the canister 22 to prevent relative rotation between the canister 22 and the filter cartridge 34.Further details relating to the interlock members 74 can be found in Application Ser.","No.","60/280,786 entitled, BOWL-CARTRIDGE FILTER HAVING INTERLOCK MECHANISM AND METHODS, which was filed on Apr.","2, 2001, and which is hereby incorporated by reference in its entirety.","Referring still to FIG.","7, the bottom wall 70 also defines a plurality of drain openings 76.As best shown in FIG.","8, each of the drain openings 76 is preferably elongated in a circumferential direction.","The drain openings 76 are positioned near the side wall 72.Inner and outer sides 78 and 80 of the drain openings 76 are defined by arcs swung about a center 82 of the bottom end cap 62.Ends 84 of the drain openings 76 are preferably rounded.","Referring to FIG.","5, the sidewall 72 of the bottom end cap 65 preferably surrounds at least a lower portion of the filter media 60.Preferably, the sidewall 72 has a height H1 that is relatively short as compared to a total height H2 of the filter media 60.In a preferred embodiment, the height H1 of the sidewall 72 is less than one-third the height H2 of the entire filter media 60.The bottom wall 70 and the sidewall 72 of the bottom end cap 65 cooperate to form a cup-like structure.","In certain embodiments (e.g., a filter assembly with a 100 millimeter diameter), a spacing gap g of approximately 5-6 millimeters is formed between the cylindrical sidewall 72 and an outer side 63 of the filter media 60.A contaminant containment chamber 94 is defined in the region located between the sidewall 72 and the filter media 60.Referring to FIG.","9, the bottom end cap 65 includes inner and outer annular flanges 86 and 88 that project upwardly from the bottom wall 70.The inner and outer flanges 86 and 88 cooperate to define an annular channel 90 for receiving the bottom end 62 of the filter media 60 (see FIG.","5).","The outer flange 88 is spaced radially inwardly from the sidewall 72 of the bottom end cap 65.A ring-shaped outer filter media 92 is preferably mounted in the region between the outer flange 88 and the sidewall 72.The drain openings 76 defined through the bottom wall 70 of the bottom end cap 65 are located directly beneath the outer filter media 92.The filter media 92 is preferably made of a material such as synthetic fibrous mesh (e.g., Syntec® brand media sold Donaldson Company, Inc.).","The filter media 92 is preferably mounted so as to pre-filter any liquid that exits the containment chamber 94 through the drain openings 76.Preferably, contaminant particles (e.g., dirt, metal, etc.)","having diameters greater 2 microns, or in the range of 2-50 microns, are retained in the chamber 94.In an alternative embodiment, particles sized larger than 0.5 microns are captured.","Still referring to FIG.","9, a lower annular extension 96 preferably projects downwardly from the bottom wall 70 of the bottom end cap 65.The lower annular extension 96 defines an inner recess 98 for mounting an annular sealing member 100 (shown in FIG.","5).","As best shown in FIG.","2, when the cartridge 34 is mounted within the canister 22, the annular sealing member 100 is adapted to provide a fluid tight seal with a lower portion 103 of the mounting post 36 located at the bottom end 32 of the canister 22.Referring once again to FIG.","9, the sidewall 72 defines a primary outer diameter.","An upper sealing element 102 projects radially outward from the side wall 72 so as to define a secondary outer diameter.","Preferably, the upper sealing element 102 is integrally formed as a single, unitary piece with the side wall 72.Preferably, the upper sealing element 102 extends circumferentially about the entire circumference of the side wall 72.As best shown in FIG.","9A, an outermost surface 104 of the upper sealing element 102 is generally rounded such that the sealing element 102 has a generally semi-circular cross-section.","Looking again to FIG.","9A, a “living hinge” 106 is defined by the sidewall 72 at a region directly below the upper sealing element 102.The term “living hinge” will be understood to mean a hinge-like structure providing a region of increased flexibility where the sidewall is predisposed to bend or flex.","For example, as shown in FIG.","9A, the living hinge 106 is formed by a region of reduced cross-sectional thickness.","For example, the hinge 106 is shown having a radial thickness T1 which is less than a primary radial thickness T2 of the sidewall 72 and is also less than a secondary radial thickness T3 of the sidewall 72.The primary radial thickness T2 is located below the living hinge 106 and the secondary radial thickness T3 is located above the living hinge 106.Preferably, the primary radial thickness T2 is greater than the secondary radial thickness T3.In one embodiment, the thickness T1 is less than or equal to one half the primary thickness T2.In another non-limiting embodiment, the thickness T1 is about 0.25 millimeter (mm), the thickness T2 is about 1 mm and the thickness T3 is about 0.5 mm.","The living hinge 106 is preferably provided by a rounded notch defined in the inner surface of the sidewall 72.The notch preferably has a generally semi-circular cross-section.","The notch preferably extends about the entire inner circumference of the sidewall 72.The portion of the sidewall 72 defining the secondary thickness T3 defines a circumferential inner surface 109 that is stepped radially outwardly relative to a corresponding inner circumferential surface 108 of the portion of the sidewall 72 defining the primary thickness T2.In use, the filter cartridge 34 is mounted within the canister 22 as shown in FIG.","2.As so mounted, the cartridge 34 fits over the core 24 and the lower annular sealing member 100 forms a fluid tight seal with the bottom end 32 of the canister 22.Also, the upper sealing element 102 of the bottom end cap 65 engages an inner surface 110 of the canister 22 to form a radial seal therewith.","The inner surface 110 of the canister 22 includes a radial taper 112 for enhancing this radial seal.","Further, as described above, the sealing members 26 and 50 form annular seals between the filter head 45 and the filter assembly 20.In use, oil from the filter head 45 enters the filter assembly 20 through openings 114.From the openings 114, the oil flows into the region between the filter media 60 and the inner surface 110 of the canister 22.The oil than flows radially inwardly through the filter media 60 and also through the perforated core 24.The filtered oil exits the filter assembly 20 by flowing upwardly through the core 24 back into the filter head 45.Contaminants filtered by the filter media 60 are retained on the outer surface of the filter media 60.Heavier contaminants move downwardly into the contamination chamber 94 defined between the sidewall 72 of the bottom end cap 65.During filtration, the upper sealing element 102 prevents contaminants from accumulating at the bottom of the canister 22 by preventing the contaminants from migrating between the sidewall 72 and the inner surface of the canister 22.Instead, the contaminants are directed into the contaminant chamber 94.When the filter media 60 becomes loaded with contaminants, the canister 22 can be unthreaded from the filter head 45 to provide access to the filter cartridge 34.After the canister 22 has been unthreaded, the filter cartridge 34 is pulled axially from the canister 22.The living hinge 106 reduces the sealing pressure that exists between the upper sealing element 102 and the inner surface of the canister 22.Thus, the filter cartridge 34 can be pulled from the canister 22 with minimal resistance.","As the filter cartridge 34 is removed from the canister 22, some of the unfiltered fluid within the containment chamber 94 flows downwardly through the annular filter 92 and is drained into the canister 22 through the drainage openings 76.The remainder of the unfiltered hydraulic fluid flows through the filter element 60 and drains into the canister 22 through a central bottom opening defined by the bottom end cap 65.Regardless of the flow path, the unfiltered hydraulic fluid within the annular flow contaminant containment chamber 94 is filtered prior to being drained into the canister 22.Consequently, only clean, filtered fluid is drained into the canister 22.The contaminants are fully contained within the chamber 94, and are discarded with the filter cartridge 34.In this manner, a new filter cartridge inserted within the canister 22 will not be contaminated by the fluid present in the canister 22.FIGS.","10-14 illustrate another bottom end cap 65′ that is an embodiment of the present invention.","The bottom end cap 65′ includes an annular flange 88′ surrounded by an outer side wall 72′.","A containment chamber 94′ is defined between the flange 88′ and the side wall 72′ (see FIG.","13).","The containment chamber 94′ is adapted for receiving an annular secondary filter such as filter 92 shown in FIG.","9.Drain openings 76′ (see FIG.","11) are defined through the bottom of the bottom end cap 65′ to allow fluid to drain from the containment chamber 94′.","Referring to FIG.","10, the upper side of the bottom end cap 65′ defines a central hub or post 200 adapted to fit inside the cylindrical filter media such as the filter media 60 shown in FIG.","3B.","Ribs 202 (shown in FIG.","11) are provided on the under side of the hub 200 for reinforcing the hub to prevent the hub from deforming in response to pressure differentials.","As shown in FIGS.","10 and 11, the bottom end cap 65′ also includes interlock members 74′ positioned at the side wall 72′.","The interlock members 74′ are adapted to fit within corresponding interlock recesses defined within a corresponding canister.","Referring still to FIGS.","10 and 11, the uppermost edge of the side wall 72′ includes a circumferential sealing element 102′ having generally the same configuration as sealing element 102.A circumferential living hinge 106′ is defined by the side wall 72′ directly below the circumferential sealing element 102′.","To further improve the flexibility of the circumferential sealing element 102′, the circumferential sealing element 102′ preferably defines at least one relief notch 206 for preventing the sealing element 102 from puckering or buckling when the bottom end cap 65′ is inserted within its corresponding canister.","In one embodiment, two of the relief notches 206 are defined through the sealing element 102′.","Preferably, the two relief notches 206 are uniformly spaced about the circumference of the sealing element 102′ so as to be approximately 180° apart.","In another embodiment, four relief notches can be defined through the sealing element 102.In this embodiment, the relief notches are preferably spaced approximately 90° apart about the circumference of the sealing element 102′.","FIG.","14 shows one representative notch configuration for the relief notches 206.In this configuration, the relief notch 206 includes two opposing, generally parallel walls that extend vertically downwardly from the uppermost portion of the sealing element 102′.","The vertical walls intersect with a transverse wall that interconnects the two vertical walls.","Thus, the depicted notch has a generally rectangular shape.","However, it will be appreciated that notches of other shapes could also be used.","In one embodiment, the spacing between the opposing walls of the notch is in the range of 0.01 to 0.05 inches, and the depth of the notch is in the range of 0.04 to 0.012 inches.","The above specification, examples and data provide a complete description of the manufacture and use of the composition of the invention.","Since many embodiments of the invention can be made without departing from the spirit and scope of the invention, the invention resides in the claims hereinafter appended."]],"string":"[\n [\n \"Filter cartridge for a filter assembly\",\n \"A filter assembly including a replaceable filter cartridge for use with a reusable filter canister.\",\n \"The filter cartridge includes a cylindrical filter media having top and bottom ends respectively mounted within top and bottom end caps.\",\n \"The bottom end cap includes a sidewall that surrounds at least a lower portion of the filter media.\",\n \"A contaminant containment chamber is defined between the sidewall and an outer side of the filter media.\",\n \"When the cartridge is mounted within the canister, the radial sealing element engages an inner surface of the canister to provide a circumferential seal.\",\n \"The sidewall also includes a living hinge positioned below the sealing element.\"\n ],\n [\n \"1.A filter cartridge comprising: a filter media having a top end and a bottom end; a top end cap mounted at the top end of the filter media; a bottom end cap mounted at the bottom end of the filter media, the bottom end cap including a bottom wall and a side wall, the side wall extending upwardly from the bottom wall and surrounding at least a lower portion of the filter element, the side wall having a top end; a circumferential sealing element that projects radially outwardly from the side wall at a location adjacent the top end of the side wall; and a circumferential living hinge defined by the sidewall at a location below the circumferential sealing element, wherein the living hinge is formed by a ring of reduced cross sectional thickness provided at the sidewall.\",\n \"2.The filter cartridge of claim 1, wherein the sidewall is spaced from an outer surface of the filter media such that a contaminant containment chamber is defined between the sidewall and the filter media.\",\n \"3.The filter cartridge of claim 2, wherein the bottom wall defines a plurality drain openings located beneath the contaminant containment chamber.\",\n \"4.The filter cartridge of claim 3, further comprising an outer filter element for filtering liquid that drains from the contaminant containment chamber through the drain holes.\",\n \"5.The filter cartridge of claim 4, wherein the outer filter element is annular and is mounted at a bottom of the contaminant containment chamber.\",\n \"6.The filter cartridge of claim 1, wherein the living hinge is defined by a circumferential notch defined within the sidewall.\",\n \"7.The filter cartridge of claim 6, wherein the notch has a generally semi-circular cross-section.\",\n \"8.The filter cartridge of claim 6, wherein the notch is defined in an inner surface of the sidewall.\",\n \"9.The filter cartridge of claim 1, further comprising at least one relief notch defined through the sealing element.\",\n \"10.The filter cartridge of claim 9, further comprising a plurality of relief notches defined through the sealing element.\",\n \"11.The filter cartridge of claim 10, wherein the relief notches are uniformly spaced about the circumference of the sealing element.\",\n \"12.The filter element of claim 11, wherein the relief notches include two relief notches spaced apart approximately 180 degrees about the circumference of the sealing element.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>Although applicable to a number of fields, the present invention relates primarily to liquid filters such as hydraulic oil filters or engine oil filters.\",\n \"Filters have long been used to remove contaminants from oil.\",\n \"It is well known that, with time, oil filters become plugged or clogged with contaminants and are required to be replaced.\",\n \"A typical oil filter assembly includes a housing (e.g., a bowl or canister) containing a cylindrical filter element.\",\n \"The cylindrical filter element typically defines a central longitudinal opening in which a central core is mounted.\",\n \"The core provides support for preventing the filter element from collapsing inward during use.\",\n \"In use, oil is directed from a system into an annular channel defined between the housing and the outer surface of the filter element.\",\n \"Pressure from the system forces the oil from the annular flow passage, through the filter element, and into the central opening of the filter element.\",\n \"From the central opening of the filter element, the filtered oil is discharged from the filter assembly.\",\n \"Disposable or “throw-away” type fluid filters are well known in the art.\",\n \"In this type of filter assembly, the filter element is permanently sealed within the housing.\",\n \"When the filter element becomes clogged with contaminants, the entire filter assembly is discarded and replaced with a new unit.\",\n \"Oil filter assemblies having replaceable filter elements are also known in the art.\",\n \"Filter assemblies with replaceable filter elements are desirable because the housing can be reused thereby reducing the amount of waste that enters the waste stream.\",\n \"Consequently, as landfill costs increase, filter assemblies having replaceable filter elements will become increasingly more desirable.\",\n \"Contamination is an important consideration relating to filter assemblies having replaceable filter elements.\",\n \"For example, prior art systems having replaceable filter elements can experience the accumulation of contaminants within the reusable housings that cover the replaceable filter elements.\",\n \"If this occurs, the “clean” side of new filter elements inserted within the contaminated housings can immediately become contaminated.\",\n \"There is a need in the art to prevent this type of contamination from occurring.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>One aspect of the present invention relates to a filter assembly including a replaceable filter cartridge for use with a reusable filter canister.\",\n \"The filter cartridge includes a cylindrical filter media having top and bottom ends respectively mounted within top and bottom end caps.\",\n \"The bottom end cap includes a sidewall that surrounds at least a lower portion of the filter media.\",\n \"A contaminant containment chamber is defined between the sidewall and an outer side of the filter media.\",\n \"A radial sealing element is located adjacent a top end of the sidewall.\",\n \"When the cartridge is mounted within the canister, the radial sealing element engages an inner surface of the canister to provide a circumferential seal.\",\n \"The sidewall also includes a living hinge positioned below the sealing element.\",\n \"In use of the filter assembly, contaminants are accumulated on the outer side of the filter element.\",\n \"Larger contaminants fall downwardly and are captured in the contaminant containment chamber defined by the lower end cap.\",\n \"The radial sealing element prevents contaminants from by-passing the contaminant containment chamber.\",\n \"When the filter element is fully loaded with contaminants, the filter cartridge can be removed from the canister and replaced.\",\n \"The living hinge facilitates removal of the filter cartridge from the canister by reducing friction between the radial sealing element and the inner surface of the canister.\",\n \"A variety of advantages of the invention will be set forth in part in the description that follows, and in part will be apparent from the description, or may be learned by practicing the invention.\",\n \"It is to be understood that both the foregoing general description and the following detailed description are illustrative and explanatory only and are not restrictive of the invention as claimed.\"\n ],\n [\n \"This application is being filed as a PCT international patent application in the name of Donaldson Company, Inc., a U.S. corporation (applicant for all designations except the U.S.), and in the names of the Stan M. Koehler, Brent A. Gulsvig, and John R. Hacker, all U.S. citizens and residents (applicants for the U.S. designation only), on Apr.\",\n \"2, 2002, designating all countries.\",\n \"FIELD OF THE INVENTION The present invention relates generally to filters.\",\n \"More particularly, the present invention relates to liquid filters (e.g., hydraulic fluid filters, engine oil filters or fuel filters) having replaceable filter elements (i.e., filter cartridges).\",\n \"BACKGROUND OF THE INVENTION Although applicable to a number of fields, the present invention relates primarily to liquid filters such as hydraulic oil filters or engine oil filters.\",\n \"Filters have long been used to remove contaminants from oil.\",\n \"It is well known that, with time, oil filters become plugged or clogged with contaminants and are required to be replaced.\",\n \"A typical oil filter assembly includes a housing (e.g., a bowl or canister) containing a cylindrical filter element.\",\n \"The cylindrical filter element typically defines a central longitudinal opening in which a central core is mounted.\",\n \"The core provides support for preventing the filter element from collapsing inward during use.\",\n \"In use, oil is directed from a system into an annular channel defined between the housing and the outer surface of the filter element.\",\n \"Pressure from the system forces the oil from the annular flow passage, through the filter element, and into the central opening of the filter element.\",\n \"From the central opening of the filter element, the filtered oil is discharged from the filter assembly.\",\n \"Disposable or “throw-away” type fluid filters are well known in the art.\",\n \"In this type of filter assembly, the filter element is permanently sealed within the housing.\",\n \"When the filter element becomes clogged with contaminants, the entire filter assembly is discarded and replaced with a new unit.\",\n \"Oil filter assemblies having replaceable filter elements are also known in the art.\",\n \"Filter assemblies with replaceable filter elements are desirable because the housing can be reused thereby reducing the amount of waste that enters the waste stream.\",\n \"Consequently, as landfill costs increase, filter assemblies having replaceable filter elements will become increasingly more desirable.\",\n \"Contamination is an important consideration relating to filter assemblies having replaceable filter elements.\",\n \"For example, prior art systems having replaceable filter elements can experience the accumulation of contaminants within the reusable housings that cover the replaceable filter elements.\",\n \"If this occurs, the “clean” side of new filter elements inserted within the contaminated housings can immediately become contaminated.\",\n \"There is a need in the art to prevent this type of contamination from occurring.\",\n \"SUMMARY OF THE INVENTION One aspect of the present invention relates to a filter assembly including a replaceable filter cartridge for use with a reusable filter canister.\",\n \"The filter cartridge includes a cylindrical filter media having top and bottom ends respectively mounted within top and bottom end caps.\",\n \"The bottom end cap includes a sidewall that surrounds at least a lower portion of the filter media.\",\n \"A contaminant containment chamber is defined between the sidewall and an outer side of the filter media.\",\n \"A radial sealing element is located adjacent a top end of the sidewall.\",\n \"When the cartridge is mounted within the canister, the radial sealing element engages an inner surface of the canister to provide a circumferential seal.\",\n \"The sidewall also includes a living hinge positioned below the sealing element.\",\n \"In use of the filter assembly, contaminants are accumulated on the outer side of the filter element.\",\n \"Larger contaminants fall downwardly and are captured in the contaminant containment chamber defined by the lower end cap.\",\n \"The radial sealing element prevents contaminants from by-passing the contaminant containment chamber.\",\n \"When the filter element is fully loaded with contaminants, the filter cartridge can be removed from the canister and replaced.\",\n \"The living hinge facilitates removal of the filter cartridge from the canister by reducing friction between the radial sealing element and the inner surface of the canister.\",\n \"A variety of advantages of the invention will be set forth in part in the description that follows, and in part will be apparent from the description, or may be learned by practicing the invention.\",\n \"It is to be understood that both the foregoing general description and the following detailed description are illustrative and explanatory only and are not restrictive of the invention as claimed.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS FIG.\",\n \"1 is an exploded, perspective view of a filter assembly constructed in accordance with the principles of the present invention, the assembly includes a canister and a replaceable filter cartridge; FIG.\",\n \"2 is an assembled view of the filter assembly of FIG.\",\n \"1 with the assembly being longitudinally bisected; FIGS.\",\n \"3A and 3B are exploded, perspective views of the filter cartridge of FIGS.\",\n \"1 and 2; FIG.\",\n \"4 is a top view of the filter cartridge of FIGS.\",\n \"3A and 3B; FIG.\",\n \"5 is a cross-sectional view taken along sectional line 5-5 of FIG.\",\n \"4; FIG.\",\n \"6 is a top, perspective view of the bottom end cap of the filter cartridge of FIGS.\",\n \"3A and 3B; FIG.\",\n \"7 is a bottom, perspective view of the bottom end cap of FIG.\",\n \"6; FIG.\",\n \"8 is bottom, plan View of the bottom end cap of FIG.\",\n \"6; FIG.\",\n \"9 is a cross-sectional view taken along section line 9-9 of FIG.\",\n \"8; FIG.\",\n \"9A is a detailed view of FIG.\",\n \"9 showing a living hinge defined by a sidewall of the bottom end cap; FIG.\",\n \"10 is a top, perspective view of an alternative bottom end cap that is an embodiment of the present invention; FIG.\",\n \"11 is a bottom, perspective view of the bottom end cap of FIG.\",\n \"10; FIG.\",\n \"12 is a top plan view of the bottom end cap of FIG.\",\n \"10; FIG.\",\n \"13 is a cross-sectional view taken along section line 13-13 of FIG.\",\n \"12; and FIG.\",\n \"14 is a cross-sectional view taken along section line 14-14 of FIG.\",\n \"12.DETAILED DESCRIPTION Reference will now be made in detail to preferred embodiments of the present invention that are illustrated in the accompanying drawings.\",\n \"Wherever possible, the same reference numbers will be used throughout the drawings to refer to the same or like parts.\",\n \"FIG.\",\n \"1 shows an embodiment of a filter assembly 20 constructed in accordance with the principles of the present invention.\",\n \"The filter assembly 20 includes a reusable canister 22 (i.e., a bowl or housing) adapted for connection to a filter head 45 (shown in FIG.\",\n \"2).\",\n \"The filter head 45 is part of a pressurized fluid/liquid system such as a fuel system, an engine lubrication system or a hydraulic oil system.\",\n \"The filter assembly 20 also includes a replaceable filter cartridge 34 that mounts within the canister 22.The filter cartridge 34 is adapted for filtering contaminants from the pressurized fluid/liquid system.\",\n \"In use, the filter assembly 20 is connected to the filter head 45.Once connected, liquid from the pressurized system is directed from the filter head 45 to an outer side of the filter cartridge 34 (see arrows 200).\",\n \"From the outer side of the cartridge 34, the liquid flows through the filter cartridge 34 to an inner “clean” side of the filter cartridge 34 (see arrows 202).\",\n \"Contaminants within the liquid are accumulated on the outer “dirty” side of the filter cartridge 34.Filtered liquid from the “clean” side of the filter cartridge 34 is re-introduced into the pressurized system (see arrow 204).\",\n \"Referring to FIG.\",\n \"1, the assembly 20 also includes a perforated core 24.As shown in FIG.\",\n \"2, the core 24 is preferably fixably mounted within the canister 22.For example, the core 24 can be press fit over a cylindrical mounting post 36 formed at a bottom end 32 of the canister 22 (see FIG.\",\n \"2).\",\n \"Adhesive can also be used to better secure the core 24 to the post 36.When assembled, as shown in FIG.\",\n \"2, the filter cartridge 34 fits over the core 24 such that the core 24 provides radial reinforcement to the filter cartridge 34.The radial reinforcement provided by the core 24 prevents the filter cartridge 34 from collapsing radially inwardly during use.\",\n \"Referring again to FIG.\",\n \"1, the filter assembly 20 further includes an annular sealing member 26 (e.g., an O-ring) and a plug 32.The annular sealing member 26 is preferably mounted adjacent a top end 28 of the canister 22 for providing a fluid tight seal with the filter head 45.The drain plug 30 is preferably threaded within a drain plug opening 31 (shown in FIG.\",\n \"2) defined by the bottom end 32 of the canister 22.The canister 22 is preferably a “spin-on” canister 22.By “spin-on”, it is meant that the canister 22 is connected to the engine head 45 by spinning (i.e., twisting, turning or rotating) the canister 22.For example, as shown in FIG.\",\n \"2, the canister 22 includes internal threads 38 that interlock with corresponding threads 47 defined by the filter head 45.The annular sealing member 26 of the canister 22 is preferably mounted within an annular recess 40 located directly beneath the threads 38.When the canister 22 is fully threaded on the filter head 45, the annular seal 26 preferably engages a cylindrical sealing surface 49 of the filter head 45 to provide a fluid tight seal.\",\n \"Still referring to FIG.\",\n \"2, the filter assembly 20 also includes an inner, annular sealing member 50.The inner, annular sealing member 50 is mounted within an annular recess 52 defined by the filter cartridge 34.When the canister 22 is fully threaded on the filter head 45, a cylindrical extension 54 of the filter head 45 projects partially into the filter cartridge 34.The extension 54 includes a cylindrical sealing surface 55 that engages the annular sealing member 50 to provide a fluid tight seal.\",\n \"Preferably, the canister 22 has a generally thin-walled construction and is formed from a metal such as aluminum, steel or other suitable material.\",\n \"The core 24 is also preferably made of a material such as steel or aluminum.\",\n \"FIGS.\",\n \"3A and 3B are exploded views of the filter cartridge 34.The filter cartridge 34 includes a cylindrical filter media 60 having top and bottom ends 61 and 62 respectively mounted within top and bottom end caps 64 and 65.A potting material (e.g., a material having adhesive characteristics such as urethane) can be used to seal and secure the top and bottom ends 61 and 62 in their respective end caps 64 and 65.The filter media 60 preferably has a pleated configuration.\",\n \"In certain embodiments, the filter media 60 can include a pleated paper media adapted for filtering liquids such as fuel, engine oil, hydraulic fluids or other liquid media.\",\n \"Exemplary types of filter media include sintered metal, cellulose, synthetic media, wire mesh, and glass fibers.\",\n \"Also, either pleated or unpleated filter media can be used.\",\n \"Referring to FIG.\",\n \"3B, the top end cap 64 of the filter cartridge 34 includes an annular channel 66 sized to receive the top end 61 of the filter media 60.The top end cap 64 also preferably includes a handle 68 (best shown in FIGS.\",\n \"3A and 4) for use in pulling the filter cartridge 34 from the canister 22 during the cartridge removal process.\",\n \"Further, the top end cap 64 preferably defines the annular recess 52 (shown in FIG.\",\n \"2) for mounting the annular sealing member 50.Referring to FIGS.\",\n \"6 and 7, the bottom end cap 65 of the filter cartridge 34 preferably includes a bottom wall 70 and a circumferential sidewall 72 that projects upwardly from the bottom wall 70.The bottom end cap 65 is preferably made of a plastic material such as glass-filled nylon.\",\n \"Preferably, the bottom end wall 70 and the sidewall 72 are integrally formed as a single unitary piece.\",\n \"In the depicted embodiment, the sidewall 72 is preferably made of a material that is impervious to liquids (e.g., hydraulic oil).\",\n \"Referring to FIG.\",\n \"7, the bottom wall 70 is preferably generally planar.\",\n \"A plurality of interlock members 74 project outwardly from the bottom side of the bottom wall 70.The interlock members 74 are adapted to engage corresponding interlock members formed in the bottom of the canister 22 to prevent relative rotation between the canister 22 and the filter cartridge 34.Further details relating to the interlock members 74 can be found in Application Ser.\",\n \"No.\",\n \"60/280,786 entitled, BOWL-CARTRIDGE FILTER HAVING INTERLOCK MECHANISM AND METHODS, which was filed on Apr.\",\n \"2, 2001, and which is hereby incorporated by reference in its entirety.\",\n \"Referring still to FIG.\",\n \"7, the bottom wall 70 also defines a plurality of drain openings 76.As best shown in FIG.\",\n \"8, each of the drain openings 76 is preferably elongated in a circumferential direction.\",\n \"The drain openings 76 are positioned near the side wall 72.Inner and outer sides 78 and 80 of the drain openings 76 are defined by arcs swung about a center 82 of the bottom end cap 62.Ends 84 of the drain openings 76 are preferably rounded.\",\n \"Referring to FIG.\",\n \"5, the sidewall 72 of the bottom end cap 65 preferably surrounds at least a lower portion of the filter media 60.Preferably, the sidewall 72 has a height H1 that is relatively short as compared to a total height H2 of the filter media 60.In a preferred embodiment, the height H1 of the sidewall 72 is less than one-third the height H2 of the entire filter media 60.The bottom wall 70 and the sidewall 72 of the bottom end cap 65 cooperate to form a cup-like structure.\",\n \"In certain embodiments (e.g., a filter assembly with a 100 millimeter diameter), a spacing gap g of approximately 5-6 millimeters is formed between the cylindrical sidewall 72 and an outer side 63 of the filter media 60.A contaminant containment chamber 94 is defined in the region located between the sidewall 72 and the filter media 60.Referring to FIG.\",\n \"9, the bottom end cap 65 includes inner and outer annular flanges 86 and 88 that project upwardly from the bottom wall 70.The inner and outer flanges 86 and 88 cooperate to define an annular channel 90 for receiving the bottom end 62 of the filter media 60 (see FIG.\",\n \"5).\",\n \"The outer flange 88 is spaced radially inwardly from the sidewall 72 of the bottom end cap 65.A ring-shaped outer filter media 92 is preferably mounted in the region between the outer flange 88 and the sidewall 72.The drain openings 76 defined through the bottom wall 70 of the bottom end cap 65 are located directly beneath the outer filter media 92.The filter media 92 is preferably made of a material such as synthetic fibrous mesh (e.g., Syntec® brand media sold Donaldson Company, Inc.).\",\n \"The filter media 92 is preferably mounted so as to pre-filter any liquid that exits the containment chamber 94 through the drain openings 76.Preferably, contaminant particles (e.g., dirt, metal, etc.)\",\n \"having diameters greater 2 microns, or in the range of 2-50 microns, are retained in the chamber 94.In an alternative embodiment, particles sized larger than 0.5 microns are captured.\",\n \"Still referring to FIG.\",\n \"9, a lower annular extension 96 preferably projects downwardly from the bottom wall 70 of the bottom end cap 65.The lower annular extension 96 defines an inner recess 98 for mounting an annular sealing member 100 (shown in FIG.\",\n \"5).\",\n \"As best shown in FIG.\",\n \"2, when the cartridge 34 is mounted within the canister 22, the annular sealing member 100 is adapted to provide a fluid tight seal with a lower portion 103 of the mounting post 36 located at the bottom end 32 of the canister 22.Referring once again to FIG.\",\n \"9, the sidewall 72 defines a primary outer diameter.\",\n \"An upper sealing element 102 projects radially outward from the side wall 72 so as to define a secondary outer diameter.\",\n \"Preferably, the upper sealing element 102 is integrally formed as a single, unitary piece with the side wall 72.Preferably, the upper sealing element 102 extends circumferentially about the entire circumference of the side wall 72.As best shown in FIG.\",\n \"9A, an outermost surface 104 of the upper sealing element 102 is generally rounded such that the sealing element 102 has a generally semi-circular cross-section.\",\n \"Looking again to FIG.\",\n \"9A, a “living hinge” 106 is defined by the sidewall 72 at a region directly below the upper sealing element 102.The term “living hinge” will be understood to mean a hinge-like structure providing a region of increased flexibility where the sidewall is predisposed to bend or flex.\",\n \"For example, as shown in FIG.\",\n \"9A, the living hinge 106 is formed by a region of reduced cross-sectional thickness.\",\n \"For example, the hinge 106 is shown having a radial thickness T1 which is less than a primary radial thickness T2 of the sidewall 72 and is also less than a secondary radial thickness T3 of the sidewall 72.The primary radial thickness T2 is located below the living hinge 106 and the secondary radial thickness T3 is located above the living hinge 106.Preferably, the primary radial thickness T2 is greater than the secondary radial thickness T3.In one embodiment, the thickness T1 is less than or equal to one half the primary thickness T2.In another non-limiting embodiment, the thickness T1 is about 0.25 millimeter (mm), the thickness T2 is about 1 mm and the thickness T3 is about 0.5 mm.\",\n \"The living hinge 106 is preferably provided by a rounded notch defined in the inner surface of the sidewall 72.The notch preferably has a generally semi-circular cross-section.\",\n \"The notch preferably extends about the entire inner circumference of the sidewall 72.The portion of the sidewall 72 defining the secondary thickness T3 defines a circumferential inner surface 109 that is stepped radially outwardly relative to a corresponding inner circumferential surface 108 of the portion of the sidewall 72 defining the primary thickness T2.In use, the filter cartridge 34 is mounted within the canister 22 as shown in FIG.\",\n \"2.As so mounted, the cartridge 34 fits over the core 24 and the lower annular sealing member 100 forms a fluid tight seal with the bottom end 32 of the canister 22.Also, the upper sealing element 102 of the bottom end cap 65 engages an inner surface 110 of the canister 22 to form a radial seal therewith.\",\n \"The inner surface 110 of the canister 22 includes a radial taper 112 for enhancing this radial seal.\",\n \"Further, as described above, the sealing members 26 and 50 form annular seals between the filter head 45 and the filter assembly 20.In use, oil from the filter head 45 enters the filter assembly 20 through openings 114.From the openings 114, the oil flows into the region between the filter media 60 and the inner surface 110 of the canister 22.The oil than flows radially inwardly through the filter media 60 and also through the perforated core 24.The filtered oil exits the filter assembly 20 by flowing upwardly through the core 24 back into the filter head 45.Contaminants filtered by the filter media 60 are retained on the outer surface of the filter media 60.Heavier contaminants move downwardly into the contamination chamber 94 defined between the sidewall 72 of the bottom end cap 65.During filtration, the upper sealing element 102 prevents contaminants from accumulating at the bottom of the canister 22 by preventing the contaminants from migrating between the sidewall 72 and the inner surface of the canister 22.Instead, the contaminants are directed into the contaminant chamber 94.When the filter media 60 becomes loaded with contaminants, the canister 22 can be unthreaded from the filter head 45 to provide access to the filter cartridge 34.After the canister 22 has been unthreaded, the filter cartridge 34 is pulled axially from the canister 22.The living hinge 106 reduces the sealing pressure that exists between the upper sealing element 102 and the inner surface of the canister 22.Thus, the filter cartridge 34 can be pulled from the canister 22 with minimal resistance.\",\n \"As the filter cartridge 34 is removed from the canister 22, some of the unfiltered fluid within the containment chamber 94 flows downwardly through the annular filter 92 and is drained into the canister 22 through the drainage openings 76.The remainder of the unfiltered hydraulic fluid flows through the filter element 60 and drains into the canister 22 through a central bottom opening defined by the bottom end cap 65.Regardless of the flow path, the unfiltered hydraulic fluid within the annular flow contaminant containment chamber 94 is filtered prior to being drained into the canister 22.Consequently, only clean, filtered fluid is drained into the canister 22.The contaminants are fully contained within the chamber 94, and are discarded with the filter cartridge 34.In this manner, a new filter cartridge inserted within the canister 22 will not be contaminated by the fluid present in the canister 22.FIGS.\",\n \"10-14 illustrate another bottom end cap 65′ that is an embodiment of the present invention.\",\n \"The bottom end cap 65′ includes an annular flange 88′ surrounded by an outer side wall 72′.\",\n \"A containment chamber 94′ is defined between the flange 88′ and the side wall 72′ (see FIG.\",\n \"13).\",\n \"The containment chamber 94′ is adapted for receiving an annular secondary filter such as filter 92 shown in FIG.\",\n \"9.Drain openings 76′ (see FIG.\",\n \"11) are defined through the bottom of the bottom end cap 65′ to allow fluid to drain from the containment chamber 94′.\",\n \"Referring to FIG.\",\n \"10, the upper side of the bottom end cap 65′ defines a central hub or post 200 adapted to fit inside the cylindrical filter media such as the filter media 60 shown in FIG.\",\n \"3B.\",\n \"Ribs 202 (shown in FIG.\",\n \"11) are provided on the under side of the hub 200 for reinforcing the hub to prevent the hub from deforming in response to pressure differentials.\",\n \"As shown in FIGS.\",\n \"10 and 11, the bottom end cap 65′ also includes interlock members 74′ positioned at the side wall 72′.\",\n \"The interlock members 74′ are adapted to fit within corresponding interlock recesses defined within a corresponding canister.\",\n \"Referring still to FIGS.\",\n \"10 and 11, the uppermost edge of the side wall 72′ includes a circumferential sealing element 102′ having generally the same configuration as sealing element 102.A circumferential living hinge 106′ is defined by the side wall 72′ directly below the circumferential sealing element 102′.\",\n \"To further improve the flexibility of the circumferential sealing element 102′, the circumferential sealing element 102′ preferably defines at least one relief notch 206 for preventing the sealing element 102 from puckering or buckling when the bottom end cap 65′ is inserted within its corresponding canister.\",\n \"In one embodiment, two of the relief notches 206 are defined through the sealing element 102′.\",\n \"Preferably, the two relief notches 206 are uniformly spaced about the circumference of the sealing element 102′ so as to be approximately 180° apart.\",\n \"In another embodiment, four relief notches can be defined through the sealing element 102.In this embodiment, the relief notches are preferably spaced approximately 90° apart about the circumference of the sealing element 102′.\",\n \"FIG.\",\n \"14 shows one representative notch configuration for the relief notches 206.In this configuration, the relief notch 206 includes two opposing, generally parallel walls that extend vertically downwardly from the uppermost portion of the sealing element 102′.\",\n \"The vertical walls intersect with a transverse wall that interconnects the two vertical walls.\",\n \"Thus, the depicted notch has a generally rectangular shape.\",\n \"However, it will be appreciated that notches of other shapes could also be used.\",\n \"In one embodiment, the spacing between the opposing walls of the notch is in the range of 0.01 to 0.05 inches, and the depth of the notch is in the range of 0.04 to 0.012 inches.\",\n \"The above specification, examples and data provide a complete description of the manufacture and use of the composition of the invention.\",\n \"Since many embodiments of the invention can be made without departing from the spirit and scope of the invention, the invention resides in the claims hereinafter appended.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10474136"}}},{"rowIdx":783,"cells":{"text":{"kind":"list like","value":[["Phage display vector, phages encoded thereby, and methods of use thereof","Filamentous bacteriophages are particularly efficient for the expression and display of combinatorial random peptides.","Two phage proteins are often employed for peptide display: the infectivity protein, pIII and the major coat protein, pVIII.","The use of pVIII typically requires the expression of two pVIII genes: the wild type and the recombinant pVIII genes to generate mosaic phages.","“Type 88” vectors contain two pVIII genes in one phage genome.","A novel “type 88” expression vector has been rationally designed and constructed, which can be used to express recombinant peptides as pVIII chimeric proteins in mosaic bacteriophages.","This vector is not only genetically stable but also of high copy number and produces high titers of recombinant phages."],["1.A DNA molecule comprising the DNA of a filamentous phage into which has been inserted a recombinant DNA sequence comprising a multiple cloning site comprising a series of restriction sites that do not otherwise appear in said DNA of the filamentous phage, said recombinant DNA sequence being designed and inserted such that the multiple cloning site is placed between a terminator for the wild type pVIII gene and an initiator for the wild type pIII gene.","2.A DNA molecule in accordance with claim 1, wherein a positive selection marker is present in the multiple cloning site.","3.A DNA molecule in accordance with claim 1, wherein an additional bacteriophage gene is present in the multiple cloning site.","4.A DNA molecule in accordance with claim 1, wherein said additional bacteriophage gene is a second pVIII gene.","5.A DNA molecule in accordance with claim 1, wherein said additional bacteriophage gene is a second pIII gene.","6.A DNA molecule in accordance with claim 1, wherein said recombinant DNA sequence is designed and inserted such that a DNA sequence encoding a foreign peptide is incorporated into the pIII gene at the N-terminal region thereof.","7.A DNA molecule in accordance with claim 1, wherein said multiple cloning site placed between said wild type pVIII gene (the first pVIII gene) and the wild type pIII gene comprises DNA encoding an additional pVIII protein (the second pVIII gene) and DNA encoding a polypeptide of interest other than pVIII.","8.A DNA molecule in accordance with claim 7, wherein the DNA encoding the second pVIII gene uses alternative codons which are other than the native codons for encoding one or more of the amino acid residues of the pVIII protein, such that no stretch of more than 30 bases of the native sequence appear therein.","9.A DNA molecule in accordance with claim 8, wherein no stretch of more than 20 bases of the native sequence appear in the DNA encoding the second pVIII gene.","10.A DNA molecule in accordance with claim 7, wherein the native pVIII DNA sequence is separated from the recombinant pVIII DNA sequence by a terminator which is not native to the filamentous phage DNA.","11.A DNA molecule in accordance with claim 7, wherein DNA encoding the native amino acid sequence of the pVIII C-terminal/pIII −35 box is maintained at the region immediately upstream to the initiation of the pIII gene.","12.A DNA molecule in accordance with claim 11, wherein said region encoding the pVIII C-terminal domain in said C-terminal/−35 box uses alternative codons which are other than the native codons, such that no stretch of more than 30 bases of the native sequence appear therein.","13.A DNA molecule in accordance with claim 12, wherein no stretch of more than 20 bases of the native sequence appear in the region encoding the pVIII C-terminal/−35 box.","14.A DNA molecule in accordance with claim 7, wherein a positive selection marker is inserted between the native and recombinant pVIII genes.","15.A DNA molecule in accordance with claim 14, wherein said positive selection marker is the tetracycline resistance gene.","16.A DNA molecule in accordance with claim 15, wherein a unidirectional promoter is substituted for the native bidirectional tetracycline resistance gene promoter.","17.A DNA molecule in accordance with claim 7, wherein the native intergenic space region is present.","18.A DNA molecule in accordance with claim 7, wherein a promoter is used for the second pVIII gene and DNA encoding the peptide of interest, which promoter is not native to the native filamentous phage.","19.A DNA molecule in accordance with claim 7, wherein said recombinant DNA sequence is designed and inserted such that a DNA sequence encoding a second peptide of interest is incorporated into the pIII gene at the N-terminal region thereof.","20.A filamentous phage encoded by the DNA of claim 7.21.A DNA molecule in accordance with claim 7 wherein said polypeptide of interest is a random peptide.","22.A library of filamentous phages encoded by a DNA molecule in accordance with claim 21, wherein said library comprises a plurality of phages each having a different random peptide as the polypeptide of interest.","23.A library in accordance with claim 22, wherein the polypeptides of interest in said plurality of phages comprise all possible random peptides of a given length or a fraction thereof which maintains the complexity thereof.","24.A filamentous phage encoded by a DNA molecule in accordance with claim 7, wherein said polypeptide of interest is a peptide which comprises a contiguous fragment of the sequence of an antigen of interest.","25.A library of filamentous phages in accordance with claim 24, wherein said library comprises a plurality of phages, said peptide in each of said plurality of phages comprising a different contiguous fragment of the sequence of the antigen of interest.","26.A library in accordance with claim 25, wherein the polypeptides of interest in said plurality of phages comprise all possible overlapping contiguous fragments of the antigen of interest.","27.A method of screening for a molecule which binds to a peptide of interest comprising bringing a molecule to be screened into contact with a filamentous phage in accordance with claim 20 which expresses said peptide of interest and, if said molecule binds to said peptide of interest, identifying and producing said molecule.","28.A method of screening for peptides which bind to a molecule of interest comprising bringing the molecule of interest into contact with a library of filamentous phages in accordance with claim 22; identifying any peptide expressed by a phage which binds to said molecule; and producing a polypeptide which comprises said peptide.","29.A method of screening for peptides which bind to a molecule of interest comprising bringing the molecule of interest into contact with a library of filamentous phages in accordance with claim 25; identifying any peptide expressed by a phage which binds to said molecule; and producing a polypeptide which comprises said peptide.","30.A DNA molecule comprising the DNA of a filamentous phage, into which has been inserted between the wild type pVIII gene (the first pVIII gene) and the wild type pIII gene, recombinant DNA sequence encoding an additional pVIII protein (the second pVIII gene) and DNA containing a region of bases flanked by two identical restriction enzyme sites, which sites do not appear anywhere else in the DNA molecule.","31.A DNA construct for insertion into the DNA of a filamentous phage, said construct comprising a multiple cloning site comprising a series of restriction sites that do not otherwise appear in the DNA of the phage into which the construct is intended to be inserted, said construct having one end compatible for ligation to a restriction enzyme site upstream of the pIII gene of the phage into which the construct is intended to be inserted and another end compatible for ligation to a restriction enzyme site downstream of the pVIII gene of the phage into which the construct is intended to be inserted.","32.A DNA construct in accordance with claim 31 further including a positive selection marker.","33.A DNA construct in accordance with claim 31 further including a second pVIII gene."],["<SOH> BACKGROUND OF THE INVENTION <EOH>Combinatorial phage display peptide libraries provide an effective means to study protein:protein interactions.","This technology relies on the production of very large collections of random peptides associated with their corresponding genetic blueprints (Scott et al, 1990; Dower, 1992; Lane et al, 1993; Cortese et al, 1994; Cortese et al, 1995; Cortese et al, 1996).","Presentation of the random peptides is often accomplished by constructing chimeric proteins expressed on the outer surface of filamentous bacteriophages such as M13, fd and f1.This presentation makes the repertoires amenable to binding assays and specialized screening schemes (referred to as biopanning (Parmley et al, 1988)) leading to the affinity isolation and identification of peptides with desired binding properties.","In this way peptides that bind to receptors (Koivunen et al, 1995; Wrighton et al, 1996; Sparks et all, 1994; Rasqualini et al, 1996), enzymes (Matthews et al, 1993; Schmitz et al, 1996) or antibodies (Scott et al, 1990; Cwirla et al, 1990; Felici et al, 1991; Luzzago et al, 1993; Hoess et al, 1993; Bonnycastle et al, 1996) have been efficiently selected.","Filamentous bacteriophages are nonlytic, male specific bacteriophages that infect Escherichia coli cells carrying an F-episome (for review, see Model et al, 1988).","Filamentous phage particles appear as thin tubular structures 900 nm long and 10 nm thick containing a circular single stranded DNA genome (the +strand).","The life cycle of the phage entails binding of the phage to the F-pilus of the bacterium followed by entry of the single stranded DNA genome into the host.","The circular single stranded DNA is recognized by the host replication machinery and the synthesis of the complementary second DNA strand is initiated at the phage ori(−) structure.","The double stranded DNA replicating form is the template for the synthesis of single-stranded DNA circular phage genomes, initiating at the ori(+) structure.","These are ultimately packaged into virions and the phage particles are extruded from the bacterium without causing lysis or apparent damage to the host.","Peptide display systems have exploited two structural proteins of the phage; pIII protein and pVIII protein.","The pIII protein exists in 5 copies per phage and is found exclusively at one tip of the virion (Goldsmith et al, 1977).","The N-terminal domain of the pIII protein forms a knob-like structure that is required for the infectivity process (Gray et al, 1981).","It enables the adsorption of the phage to the tip of the F-pilus and subsequently the penetration and translocation of the single stranded phage DNA into the bacterial host cell (Holliger et al, 1997).","The pIII protein can tolerate extensive modifications and thus has been used to express peptides at its N-terminus.","The foreign peptides have been up to 65 amino acid residues long (Bluthner et al, 1996; Kay et al, 1993) and in some instances even as large as full-length proteins (McCafferty et al, 1990; McCafferty et al, 1992) without markedly affecting pIII function.","The cylindrical protein envelope surrounding the single stranded phage DNA is composed of 2700 copies of the major coat protein, pVIII, an α-helical subunit which consists of 50 amino acid residues.","The pVIII proteins themselves are arranged in a helical pattern, with the α-helix of the protein oriented at a shallow angle to the long axis of the virion (Marvin et al, 1994).","The primary structure of this protein contains three separate domains: (1) the N-terminal part, enriched with acidic amino acids and exposed to the outside environment; (2) a central hydrophobic domain responsible for: (i) subunit:subunit interactions in the phage particle and (ii) transmembrane functions in the host cell; and (3) the third domain containing basic amino acids, clustered at the C-terminus, which is buried in the interior of the phage and is associated with the phage-DNA.","pVIII is synthesized as a precoat protein containing a 23 amino acid leader-peptide, which is cleaved upon translocation across the inner membrane of the bacterium to yield the mature 50-residue transmembrane protein (Sugimoto et al, 1977).","Use of pVIII as a display scaffold is hindered by the fact that it can tolerate the addition of peptides no longer than 6 residues at its N-terminus (Greenwood et al, 1991; Iannolo et al, 1995).","Larger inserts interfere with phage assembly.","Introduction of larger peptides, however, is possible in systems where mosaic phages are produced by in vivo mixing the recombinant, peptide-containing, pVIII proteins with wild type pVIII (Felici et al, 1991; Greenwood et al, 1991; Willis et al, 1993).","This enables the incorporation of the chimeric pVIII proteins at low density (tens to hundreds of copies per particle) on the phage surface interspersed with wild type coat proteins during the assembly of phage particles.","Two systems have been used that enable the generation of mosaic phages; the “type 8+8” and “type 88” systems as designated by Smith (Smith, 1993).","The “type 8+8” system is based on having the two pVIII genes situated separately in two different genetic units (Felici et al, 1991; Greenwood et al, 1991; Willis et al, 1993).","The recombinant pVIII gene is located on a phagemid, a plasmid that contains, in addition to its own origin of replication, the phage origins of replication and packaging signal.","The wild type pVIII protein is supplied by superinfecting phagemid-harboring bacteria with a helper phage.","In addition, the helper phage provides the phage replication and assembly machinery that package both the phagemid and the helper genomes into virions.","Therefore, two types of particles are secreted by such bacteria, helper and phagemid, both of which incorporate a mixture of recombinant and wild type pVIII proteins.","The “type 88” system benefits by containing the two pVIII genes in one and the same infectious phage genome.","Thus, this obviates the need for a helper phage and superinfection.","Furthermore, only one type of mosaic phage is produced.","The question arises, however, where one should introduce the second pVIII gene within the filamentous phage genome for efficient expression and genetic stability.","The phage genome encodes 10 proteins (pI through pX) all of which are essential for production of infectious progeny (Felici et al, 1991).","The genes for the proteins are organized in two tightly packed transcriptional units separated by two non-coding regions (Van Wezenbeek et al, 1980).","One non-coding region, called the “intergenic region” (defined as situated between the pIV and pII genes) contains the (+) and the (−) origins of DNA replication and the packaging signal of the phage, enabling the initiation of capsid formation.","Parts of this intergenic region are dispensable (Kim et al, 1981; Dotto et al, 1984).","Moreover, this region has been found to be able to tolerate the insertion of foreign DNAs at several sites (Messing, 1983; Moses et al, 1980; Zacher et al, 1980).","The second non-coding region of the phage is located between the pVIII and pIII genes, and has also been used to incorporate foreign recombinant genes as was illustrated by Pluckthun (Krebber et al, 1995).","Regardless as to where a second pVIII gene is to be introduced, a major point for concern is the genetic stability of the ultimate vector and its derivatives."],["<SOH> SUMMARY OF THE INVENTION <EOH>The present invention is based on a critical examination of the attributes of the two non-coding regions of the fd filamentous phage as potential sites for insertion of a second recombinant pVIII gene and its genetic stability, resulting in the design and construction of an efficient “type 88” phage display expression system.","The phage display expression system of the present invention includes a vector which is the DNA sequence of a filamentous phage into which a second pVIII gene, as well as DNA encoding a peptide of interest, are placed between the wild type pVIII and pIII genes.","This allows production of type 88 phages displaying a peptide of interest with genetic stability and high copy number.","The DNA encoding the second pVIII gene preferably uses alternative codons for encoding the amino acid residues of the pVIII protein.","The native pVIII DNA sequence is separated from the recombinant pVIII sequence by a region encoding the wild type pVIII C-terminal domain and a terminator.","The region encoding the wild type pVIII C-terminal domain is preferably designed to use alternative codons which are other than the native codons for encoding the same amino acids.","In this way homologous recombination, slippage mechanisms and other genetic instabilities may be avoided.","Preferably, the terminator at the end of the native pVIII DNA sequence is an HP terminator which is not native to the filamentous phage DNA.","In a preferred embodiment of the present invention, a positive selection marker, such as the tetracycline or kanamycin resistant genes, is inserted between the native and recombinant pVIII genes.","Preferably, a unidirectional promoter is substituted for the native bidirectional tetracycline resistant gene promoter.","In preferred embodiments of the present invention, the native intergenic space between pIV and pII is maintained.","The present invention further relates to the filamentous phages encoded by the DNA discussed above.","Such phages may be of any type, such as fd, M13 and f1, although fd is preferred.","The peptide designed to be displayed by the phage of the present invention can be any peptide which is desired to be presented, such as a specific antigen.","Indeed, any protein or peptide can be displayed on the phage of the present invention.","The peptide displayed on the phage may be an epitope of an antigen, which phage can be used therapeutically as a vaccine.","Furthermore, the peptide displayed on the phage may be a single-chain antibody, which phage can be used for passive immunization.","Both of these embodiments are described in WO 01/18169.Alternatively, one can create a library of phages into which are incorporated, at the same site, each of a set of oligonucleotides that encode all possible random peptides of a given length, or a large subset of that set.","Such a large subset should preferably represent a fraction of the full set with the theoretical complexity of the full set of random polypeptides of a given length.","Libraries of phages can also be prepared which incorporate all, or a large subset of all, of the overlapping oligonucleotides that represent all of the overlapping peptides of a given antigen and, thus, create a phage display pepscan.","Scrambled pepscans (PCT application no.","WO 98/20169) and phage display two hybrid systems (PCT application no.","WO 98/20159) may also be prepared by this technique.","The phage display libraries of the present invention can be used in screening for molecules which bind to a particular displayed peptide of interest or in screening the peptides displayed on a library of phages to see which bind to a specific molecule of interest, all as is well known with respect to prior art phage display libraries.","Once a molecule or peptide of interest is identified by means of the screen using the phages or phage display libraries of the present invention, the peptide or molecule so identified may be produced in a conventional manner."],["FIELD OF THE INVENTION The present invention is related to a phage display vector encoding a filamentous phage which includes DNA encoding a polypeptide of interest and, more particularly, to such a vector in which a recombinant sequence including a multiple cloning site is inserted between the wild type pVIII and pIII genes.","The invention further relates to the phages encoded by such DNA, phage display libraries made of such phages and methods of use thereof.","BACKGROUND OF THE INVENTION Combinatorial phage display peptide libraries provide an effective means to study protein:protein interactions.","This technology relies on the production of very large collections of random peptides associated with their corresponding genetic blueprints (Scott et al, 1990; Dower, 1992; Lane et al, 1993; Cortese et al, 1994; Cortese et al, 1995; Cortese et al, 1996).","Presentation of the random peptides is often accomplished by constructing chimeric proteins expressed on the outer surface of filamentous bacteriophages such as M13, fd and f1.This presentation makes the repertoires amenable to binding assays and specialized screening schemes (referred to as biopanning (Parmley et al, 1988)) leading to the affinity isolation and identification of peptides with desired binding properties.","In this way peptides that bind to receptors (Koivunen et al, 1995; Wrighton et al, 1996; Sparks et all, 1994; Rasqualini et al, 1996), enzymes (Matthews et al, 1993; Schmitz et al, 1996) or antibodies (Scott et al, 1990; Cwirla et al, 1990; Felici et al, 1991; Luzzago et al, 1993; Hoess et al, 1993; Bonnycastle et al, 1996) have been efficiently selected.","Filamentous bacteriophages are nonlytic, male specific bacteriophages that infect Escherichia coli cells carrying an F-episome (for review, see Model et al, 1988).","Filamentous phage particles appear as thin tubular structures 900 nm long and 10 nm thick containing a circular single stranded DNA genome (the +strand).","The life cycle of the phage entails binding of the phage to the F-pilus of the bacterium followed by entry of the single stranded DNA genome into the host.","The circular single stranded DNA is recognized by the host replication machinery and the synthesis of the complementary second DNA strand is initiated at the phage ori(−) structure.","The double stranded DNA replicating form is the template for the synthesis of single-stranded DNA circular phage genomes, initiating at the ori(+) structure.","These are ultimately packaged into virions and the phage particles are extruded from the bacterium without causing lysis or apparent damage to the host.","Peptide display systems have exploited two structural proteins of the phage; pIII protein and pVIII protein.","The pIII protein exists in 5 copies per phage and is found exclusively at one tip of the virion (Goldsmith et al, 1977).","The N-terminal domain of the pIII protein forms a knob-like structure that is required for the infectivity process (Gray et al, 1981).","It enables the adsorption of the phage to the tip of the F-pilus and subsequently the penetration and translocation of the single stranded phage DNA into the bacterial host cell (Holliger et al, 1997).","The pIII protein can tolerate extensive modifications and thus has been used to express peptides at its N-terminus.","The foreign peptides have been up to 65 amino acid residues long (Bluthner et al, 1996; Kay et al, 1993) and in some instances even as large as full-length proteins (McCafferty et al, 1990; McCafferty et al, 1992) without markedly affecting pIII function.","The cylindrical protein envelope surrounding the single stranded phage DNA is composed of 2700 copies of the major coat protein, pVIII, an α-helical subunit which consists of 50 amino acid residues.","The pVIII proteins themselves are arranged in a helical pattern, with the α-helix of the protein oriented at a shallow angle to the long axis of the virion (Marvin et al, 1994).","The primary structure of this protein contains three separate domains: (1) the N-terminal part, enriched with acidic amino acids and exposed to the outside environment; (2) a central hydrophobic domain responsible for: (i) subunit:subunit interactions in the phage particle and (ii) transmembrane functions in the host cell; and (3) the third domain containing basic amino acids, clustered at the C-terminus, which is buried in the interior of the phage and is associated with the phage-DNA.","pVIII is synthesized as a precoat protein containing a 23 amino acid leader-peptide, which is cleaved upon translocation across the inner membrane of the bacterium to yield the mature 50-residue transmembrane protein (Sugimoto et al, 1977).","Use of pVIII as a display scaffold is hindered by the fact that it can tolerate the addition of peptides no longer than 6 residues at its N-terminus (Greenwood et al, 1991; Iannolo et al, 1995).","Larger inserts interfere with phage assembly.","Introduction of larger peptides, however, is possible in systems where mosaic phages are produced by in vivo mixing the recombinant, peptide-containing, pVIII proteins with wild type pVIII (Felici et al, 1991; Greenwood et al, 1991; Willis et al, 1993).","This enables the incorporation of the chimeric pVIII proteins at low density (tens to hundreds of copies per particle) on the phage surface interspersed with wild type coat proteins during the assembly of phage particles.","Two systems have been used that enable the generation of mosaic phages; the “type 8+8” and “type 88” systems as designated by Smith (Smith, 1993).","The “type 8+8” system is based on having the two pVIII genes situated separately in two different genetic units (Felici et al, 1991; Greenwood et al, 1991; Willis et al, 1993).","The recombinant pVIII gene is located on a phagemid, a plasmid that contains, in addition to its own origin of replication, the phage origins of replication and packaging signal.","The wild type pVIII protein is supplied by superinfecting phagemid-harboring bacteria with a helper phage.","In addition, the helper phage provides the phage replication and assembly machinery that package both the phagemid and the helper genomes into virions.","Therefore, two types of particles are secreted by such bacteria, helper and phagemid, both of which incorporate a mixture of recombinant and wild type pVIII proteins.","The “type 88” system benefits by containing the two pVIII genes in one and the same infectious phage genome.","Thus, this obviates the need for a helper phage and superinfection.","Furthermore, only one type of mosaic phage is produced.","The question arises, however, where one should introduce the second pVIII gene within the filamentous phage genome for efficient expression and genetic stability.","The phage genome encodes 10 proteins (pI through pX) all of which are essential for production of infectious progeny (Felici et al, 1991).","The genes for the proteins are organized in two tightly packed transcriptional units separated by two non-coding regions (Van Wezenbeek et al, 1980).","One non-coding region, called the “intergenic region” (defined as situated between the pIV and pII genes) contains the (+) and the (−) origins of DNA replication and the packaging signal of the phage, enabling the initiation of capsid formation.","Parts of this intergenic region are dispensable (Kim et al, 1981; Dotto et al, 1984).","Moreover, this region has been found to be able to tolerate the insertion of foreign DNAs at several sites (Messing, 1983; Moses et al, 1980; Zacher et al, 1980).","The second non-coding region of the phage is located between the pVIII and pIII genes, and has also been used to incorporate foreign recombinant genes as was illustrated by Pluckthun (Krebber et al, 1995).","Regardless as to where a second pVIII gene is to be introduced, a major point for concern is the genetic stability of the ultimate vector and its derivatives.","SUMMARY OF THE INVENTION The present invention is based on a critical examination of the attributes of the two non-coding regions of the fd filamentous phage as potential sites for insertion of a second recombinant pVIII gene and its genetic stability, resulting in the design and construction of an efficient “type 88” phage display expression system.","The phage display expression system of the present invention includes a vector which is the DNA sequence of a filamentous phage into which a second pVIII gene, as well as DNA encoding a peptide of interest, are placed between the wild type pVIII and pIII genes.","This allows production of type 88 phages displaying a peptide of interest with genetic stability and high copy number.","The DNA encoding the second pVIII gene preferably uses alternative codons for encoding the amino acid residues of the pVIII protein.","The native pVIII DNA sequence is separated from the recombinant pVIII sequence by a region encoding the wild type pVIII C-terminal domain and a terminator.","The region encoding the wild type pVIII C-terminal domain is preferably designed to use alternative codons which are other than the native codons for encoding the same amino acids.","In this way homologous recombination, slippage mechanisms and other genetic instabilities may be avoided.","Preferably, the terminator at the end of the native pVIII DNA sequence is an HP terminator which is not native to the filamentous phage DNA.","In a preferred embodiment of the present invention, a positive selection marker, such as the tetracycline or kanamycin resistant genes, is inserted between the native and recombinant pVIII genes.","Preferably, a unidirectional promoter is substituted for the native bidirectional tetracycline resistant gene promoter.","In preferred embodiments of the present invention, the native intergenic space between pIV and pII is maintained.","The present invention further relates to the filamentous phages encoded by the DNA discussed above.","Such phages may be of any type, such as fd, M13 and f1, although fd is preferred.","The peptide designed to be displayed by the phage of the present invention can be any peptide which is desired to be presented, such as a specific antigen.","Indeed, any protein or peptide can be displayed on the phage of the present invention.","The peptide displayed on the phage may be an epitope of an antigen, which phage can be used therapeutically as a vaccine.","Furthermore, the peptide displayed on the phage may be a single-chain antibody, which phage can be used for passive immunization.","Both of these embodiments are described in WO 01/18169.Alternatively, one can create a library of phages into which are incorporated, at the same site, each of a set of oligonucleotides that encode all possible random peptides of a given length, or a large subset of that set.","Such a large subset should preferably represent a fraction of the full set with the theoretical complexity of the full set of random polypeptides of a given length.","Libraries of phages can also be prepared which incorporate all, or a large subset of all, of the overlapping oligonucleotides that represent all of the overlapping peptides of a given antigen and, thus, create a phage display pepscan.","Scrambled pepscans (PCT application no.","WO 98/20169) and phage display two hybrid systems (PCT application no.","WO 98/20159) may also be prepared by this technique.","The phage display libraries of the present invention can be used in screening for molecules which bind to a particular displayed peptide of interest or in screening the peptides displayed on a library of phages to see which bind to a specific molecule of interest, all as is well known with respect to prior art phage display libraries.","Once a molecule or peptide of interest is identified by means of the screen using the phages or phage display libraries of the present invention, the peptide or molecule so identified may be produced in a conventional manner.","BRIEF DESCRIPTION OF THE DRAWINGS FIGS.","1A and 1B are a comparison between fd (FIG.","1A) and fd-tet (FIG.","1B).","Opened arrows indicate the phage-transcriptional units.","The darkened segment in FIG.","1B indicates the tet fragment introduced into the BamH I site of fd at position 6272 (bold in FIG.","1A) in the intergenic region (IG).","FIGS.","2A-2B show a deletion of 2.8 kb in fd-tet.","FIG.","2A is an uncut double-stranded DNA of fd-tet isolated from K802 (lane 1) or K91KAN (lane 2) and run on 0.8% agarose gel.","Uncut double-stranded DNA of wild type fd was used for comparison (lane 3).","M: DNA ladder mix (numbers equal base pairs).","FIG.","2B shows Msc I digests of fd-tet derived from K802 (lane 1) or K91KAN (lane 2) which were run on 0.8% agarose gel.","Msc I digest of fd was used for comparison (lane 3).","M: λ Hind III digest (numbers equal base pairs).","Note that the lower band in lane 2 in FIG.","2B, corresponding to the deleted product, is at least as intense as the upper band corresponding to the linear full-length fd-tet .","FIG.","3 shows the genetic instability of the inserted tet fragment.","SnaB I/Ava I digests of fd-tet derived from K802 (lane 1) or K91KAN (lane 2).","SnaB I/Ava I digest of fd was used for comparison (lane 3).","BspH I/Hind III digests of fd-tet derived from K802 (lane 4) or K91KAN (lane 5:).","BspH I/Hind III digest of fd DNA was used for comparison (lane 6).","The arrow indicates the deleted products (lane 2 and 5) corresponding in size to the linear full-length fd (lane 3 and 6).","M: DNA ladder mix.","Agarose gel concentration was 0.8%.","FIGS.","4A-4B show that the deletion is not a precise reversion to fd.","FIG.","4A is BamH I digests of fd-tet DNA derived from K802 (lane 1) or K91KAN (and 2).","BamH I digest of fd was used for comparison (lane 3).","Note that the pattern obtained in lane 2 is identical to the Msc I digest in FIG.","2B, lane 2.M: DNA ladder mix.","Agarose gel concentration was 0.8%.","FIG.","4B is BstY I digests of fd-tet derived from K802 (lane 1) or K91KAN (lane 2) run on 0.8% agarose gel.","BstY I digest of fd was used for comparison (lane 3).","M: DNA ladder mix.","Note the relative drop in intensity for the lowest band in FIG.","4B, lane 2.FIG.","5 shows the genome of the fd-tet phage.","Genes (darkened segments), promoters (bent arrows), terminators (stem and loop), and selected restriction sites relevant to this study are shown.","Strong promoters and terminators are in black.","Weak promoters and terminators are in gray.","The wild type fd genes (pI through pX) are packed in two transcriptional units.","The inserted tet fragment contains two genes; tetA encoding for the tetracycline resistance protein and tetR encoding for the repressor protein regulating the expression of the tetA gene.","The overlapping promoter/terminator, between pVIII and pIII genes, are inseparable.","Moreover, the −35 box of this promoter is situated in the C-terminal coding region of pVIII gene (see text).","Note the bi-directional promoter driving tetR and teta transcription.","FIGS.","6A-6C show the construction of two tandem pVIII genes.","FIG.","6A is a detailed comparison between the wild type pVIII gene (the contiguous central portion being SEQ ID NO:1) and the recombinant pVIII gene (the contiguous central portion being SEQ ID NO:3), designated pVIIISTS.","The amino acid sequence shown to be encoded by SEQ ID NO:1 is SEQ ID NO:2.The amino acid sequences shown to be encoded by SEQ ID NO:3 are SEQ ID NOs:4 and 5.In the pVIII gene, the GAC codon for Asp at position 4 of the wild type pVIII is deleted and replaced by a sequence of 62 bp containing two stop codons and trpA transcription terminator flanked by two Sfi I sites (“STS” insert) (nucleotides 13-74 of SEQ ID NO:3).","FIG.","6B is the pGEM-t(p8STS) construct.","The fragment containing the pVIIISTS gene, preceded by a SnaB I restriction site and followed by downstream pIII gene, was introduced into the pGEM-T vector.","Black segments are sequences corresponding to the pGEM-T vector.","The 62 bp “STS” insert is indicated.","FIG.","6C is the pGEM-t(pB8STS) construct.","This construct contains the wild type pVIII and the modified pVIIISTS genes arranged as tandem repeats.","The fragment containing the wild type pVIII gene starting from the SnaB I site and ending just beyond the overlapping promoter/terminator was introduced into the SnaB I site of pGEM-T(p8STS) thus destroying the former site while concomitantly introducing a new SnaB I site upstream to the wild type pVIII gene.","Furthermore, additional unique sites were introduced between the two pVIII genes.","FIG.","7 is the genome of ftac88 vector.","Most of the tetR gene was exchanged for a linker containing the Ava II site, thus eliminating the second SnaB I site previously situated in this region (for comparison see FIG.","5).","The modified pVIIISTS gene is situated between the duplicated overlapping promoter/terminator structures, without disrupting the two-phage transcriptional units.","Note the pVIIISTS gene is under the control of a tac promoter (Ptac).","FIG.","8 shows the binding of mAb GV4H3 to phages expressing the GV4H3 epitope.","Equal amounts of phages were applied in duplicates to a nitrocellulose membrane filter and reacted with the GV4H3 mAb.","The GV4H3 epitope displaying phages, ftac88(4H3) and fth1(4H3), showed strong signals.","Phage ftac88 and fth1 were used as negative controls.","FIG.","9 shows the deletion of the recombinant pVIII gene in ftac88(4H3).","SnaB I/BamH I digests of ftac88 derived from DH5α (lane 1) or DH5αF′ (lane 2) were run on 0.8% agarose gel.","SnaB I/BamH I digests of ftac88 (4H3) isolated from DH5α (lane 3) or DH5αF′ (lane 4) are also shown.","M: DNA ladder mix.","The upper arrow indicates the tet insert deleted product and the lower arrow indicates the recombinant pVIII gene deleted fragment.","FIG.","10 shows the fth1 vector (SEQ ID NO:29).","The general scheme of the fth1 vector is shown in scale illustrating the phage genes (opened segments), the reconstituted intergenic region (IG) and the genes introduced between the wild type pVIII and pIII genes (gray segments).","Below, detail of the pIX to the middle of pIII is provided (not in scale).","The darkened segments represent the modified wild type pVIII 3′-end and the −35 box of the pIII promoter (see also FIG.","11).","For details see text.","FIGS.","11A-11C show the intermediate vectors constructed for the formation of fth1 vector.","FIG.","11A is a detail of the segment of fd-tet containing the pIX, pVIII, and pIII genes.","The segment in pVIII indicating the −35 box of the pIII promoter is shown in black situated in the 3′ region of the wild type pVIII open reading frame.","FIG.","11B is the intermediate vector-1 (IV-1).","The −35 box of the pIII promoter was separated from the wild type pVIII gene by inserting a small segment reconstituting the 3′ region and introducing multiple cloning sites.","FIG.","11C is IV-4.A novel HP terminator (tHP) is situated downstream to the wild type pVIII gene terminating its transcription followed by the tetracycline resistance gene (tetA) under the control of the kan promoter (Pkan).","FIG.","12 shows the genetic stability of fth1.BstN I digests of fth1 derived from DH5α (lane 2) or DH5αF′ (lane 3).","BstN I digests of fth1(4H3) derived from DH5α (lane 4) or DH5αF′ (lane 5).","BstN I digest of fd was used for comparison (lane 1).","M: DNA ladder mix.","Agarose gel concentration was 0.8%.","DETAILED DESCRIPTION OF THE INVENTION Type 88 display systems are in essence a phage genome that contains two protein VIII genes.","One of the genes is the wild type pVIII, whereas the second is modified so as to enable expression of a peptide (random or pre-defined peptide, or even a single-chain antibody (scFv)).","The problem with known type 88 phage display systems is genetic instability.","After a number of generations, the phages tend to stop production of the inserted polypeptide of interest and no longer display such polypeptide.","The novel constructs of the present invention solve this problem.","Furthermore, in prior art systems, in which the selectable marker has been introduced into the intergenic region, some of the functions of the replication of the phage are obstructed, and the actual amount of phage DNA being produced in a liter of culture may be no more than 20-50 μg of DNA.","However, with the construct of the present invention, high copy numbers may be obtained.","Thus, as much as 500-1000 μg of DNA can be readily purified from a liter of culture.","In the vector of the present invention, the sequence of a filamentous phage is manipulated so as to insert a second pVIII gene and the gene encoding the polypeptide of interest between the wild type pVIII and pIII genes.","In a preferred embodiment, the intergenic region between pII and pIV is not modified and retains the native sequence, thus allowing for the high copy number to be obtained.","Any positive selection marker may also be introduced between the wild type pVIII and pIII genes.","In the wild type filamentous phage DNA, the −35 box necessary for pIII expression is positioned to overlap with the C-terminal coding region of the pVIII protein.","Thus, the −35 regulatory sequence is upstream of the C-terminal coding region and terminator of the pVIII protein.","So as not to disrupt the expression of the pIII protein, the vector of the present invention preferably maintains the genuine C-terminal/−35 box at the 5′ end of the pIII gene.","A new C-terminal region for the native pVIII gene is introduced upstream thereof.","The second pVIII region and the gene encoding the polypeptide of interest is introduced between the newly-introduced terminator for the native pVIII gene and the C-terminal/−35 box 5′ to the pIII gene.","In order to prevent possible homologous recombination, slippage mechanisms, or other genetic instabilities, it is preferable that the DNA encoding the second pVIII gene use alternative codons which are other than the native codons for encoding amino acid residues of the pVIII protein.","Enough alternative codons should be inserted therein so as to avoid stretches longer than 30 bases of the native sequence, preferably no longer than 20 such bases.","Similarly, the region encoding the wild type pVIII C-terminal domain should also use alternative codons in sufficient quantity to avoid stretches longer than 30 native bases, and preferably to avoid stretches longer than 20 such bases.","The positive selection marker is preferably inserted between the native and recombinant pVIII genes.","The present invention further comprehends a vector in which the second pVIII gene is introduced just downstream to the −35 promotor of the pIII gene (and thus driven by the pIII promotor and not the tac promotor of fth1 or ftac88 as described).","In such a situation, due to the presence of the transcription terminator in the STS stuffer of the recombinant pVIII, phages that do not incorporate an insert for the polypeptide of interest will not be produced at all.","This is due to the fact that the terminator prevents the expression of the pIII, pVI and pI proteins.","The pI protein is essential for phage assembly.","In the situation where an insert is cloned in place of the stuffer, the recombinant phage is made.","If such an insert is not incorporated, then transcription is terminated and no phage is produced.","While the fd filamentous phage is used in the present examples and is the preferred phage sequence for use in the present invention, it should be understood that all filamentous phages are very similar and have the same gene organization (Model et al, 1988).","Thus, the principles of the present invention can be applied to any of the filamentous phages, such as M13, f1 and others.","The native pVIII DNA sequence is preferably separated from the second pVIII DNA sequence by a terminator which is not native to the filamentous phage DNA, preferably the HP terminator.","In the preferred embodiment of the present invention, the native, unique, overlapping pVIII terminator/pIII promoter is maintained at its native position at the N-terminal portion of the pIII gene.","This includes the pVIII C-terminal region/pIII −35 box.","However, to further avoid tandem repeats, the pVIII C-terminal region, which is present at this point, is preferably designed with alternative codon usage to avoid stretches of longer than 30 native bases, and preferably to avoid stretches longer than 20 native bases.","While the DNA molecule of the present invention is described as having a recombinant DNA sequence encoding the pVIII protein and DNA encoding a polypeptide of interest other than pVIII which is inserted between the wild type pVIII and pIII genes, this language encompasses the presence of DNA encoding a peptide of interest within the sequence encoding the pVIII protein.","Preferably, the DNA sequence encoding the peptide of interest is present at the N-terminal region of the pVIII gene.","The peptide of interest and the pVIII gene will be expressed as a fusion protein in which the peptide of interest is expressed on the surface of the filamentous phage.","While the selectable marker disclosed herein is the tetracycline resistance gene, those of ordinary skill in the art will understand that any selectable marker can be used for this purpose, such as the kanamycin resistance gene.","Furthermore, as the native promoter for the tetracycline resistance gene is bidirectional, in a preferred embodiment of the present invention, this promoter is replaced by a unidirectional promoter, such as the kanamycin promoter.","The present invention also comprehends a vector which may be used as an intermediate for producing a vector containing the DNA encoding the peptide of interest.","In this intermediate vector, the DNA of the phage has inserted therein a recombinant DNA sequence that includes a multiple cloning site.","The multiple cloning site has a series of restriction sites that do not otherwise appear in the DNA of the phage into which the recombinant DNA has been inserted.","The recombinant DNA sequence is designed and inserted such that the multiple cloning site is placed between a terminator for the wild type pVIII gene and an initiator for the wild type pII gene.","The multiple cloning site may then be used to readily insert DNA sequences encoding the foreign peptide of interest to be displayed on the phage.","A positive selection marker, as has been discussed elsewhere in the present specification, may be present in the multiple cloning site in order to simplify insertion only of the DNA encoding the peptide of interest so as to arrive at the final product.","The second pVIII gene may also be present in the multiple cloning site.","Instead of a second pVIII gene, any recombinant bacteriophage gene can be present in the multiple cloning site.","For example, it would be useful to produce a system in which two pIII genes are present.","This would allow the production of both wild type and recombinant pIII genes in the same phage.","It is expected that, by modifying the efficiency of expression, one can create a situation in which only one recombinant pIII protein is expressed per phage.","The normal situation is that five pIII proteins are present per phage.","When all are recombinant, then the binding is affected by the multivalency of the five copies which leads to enhanced avidity of binding and can be misleading if one wants to be precise in evaluating real affinity (as opposed to avidity which benefits from multiple binding reactions).","Similarly, any of the native proteins (pI through pX) can be duplicated in the multiple cloning site for whatever reason.","The recombinant DNA sequence, which is inserted into the DNA of the filamentous phage, is designed and inserted such that the multiple cloning site is placed between a terminator for the wild type pVIII gene and an initiator for the wild type pIII gene.","As discussed hereinabove, preferably the wild type C-terminal/−35 box is retained at the 5′ end of the pIII gene, and a new C-terminal region for the native pVIII gene is introduced upstream of the multiple cloning site.","Preferably, in the intermediate vector, in the region between flanking restriction enzyme sites, a trpA transcription terminator is disposed, although any random combination of bases can appear between these flanking restriction enzyme sites.","When this intermediate is to be used for production of the vector to be used to make a phage displaying the peptide of interest, DNA encoding the peptide of interest is merely substituted for the DNA of the intermediate vector between the two flanking unique restriction enzyme sites.","Since the recombinant DNA sequence inserted into the phage in order to create the multiple cloning site must include a region homologous to the N-terminal portion of the pIII gene, it is possible in this same insert to include a DNA sequence encoding another foreign peptide incorporated into the pIII gene at the N-terminal region thereof.","This may be used, for example, to produce a phage that displays a library of random peptides on either pIII or pVIII and a marker on the other, such as green fluorescence protein.","This would make the selection of the bound phage easier or amenable to high throughput screening.","Also encompassed by the present invention is the recombinant DNA construct which is produced and used for insertion of the multiple cloning sites, into the phage DNA so as to appear between the pVIII and pIII genes.","This construct is designed so as to replace the wild type sequence between restriction sites which are already present in the phage, such as the SnaBI in the pIX gene and the BamHI in the pIII gene.","The ends of the construct are designed so as to be compatible with the restriction enzyme cleavage sites left after removal of the wild type sequence such that efficient ligation of the construct into the vector may be obtained, all as is well known to those of ordinary skill in the art.","The novel DNA vectors of the present invention and the novel type 88 filamentous phages encoded thereby may be used in the same manner that known peptide display phages have been known to be used in the prior art, except that they are genetically stable, have high copy number, and produce high titres of phages.","The vector can be used to express any protein or peptide.","In the situation in which a discrete sequence of peptide is known which is desired to be expressed, then such a known peptide can readily be incorporated into the vector so as to produce high titres of phage displaying the desired discrete amino acid sequence.","An example of the use of a discrete sequence of peptide displayed on a phage is represented by the disclosure of PCT application no.","WO 01/18169, the entire contents of which are hereby incorporated herein by reference.","This publication discloses the utility as a vaccine of phages displaying a particular antigenic epitope so as to raise antibodies against that epitope.","Also disclosed therein are phages displaying single-chain antibodies which can be directly used for passive immunization.","Thus, phages displaying such antigens or single-chain antibodies for use in the particular applications of WO 01/18169, or any related application, are considered to be comprehended by the present invention when the antigen or single-chain antibody is displayed on a phage in accordance with the present invention.","Additionally, as is known in the art, libraries can be prepared that incorporate into the same site of the vector a set of oligonucleotides that code all possible random peptides of a given length, or a large subset of that set, as is known in the art.","Such a large subset should preferably represent a fraction of the full set with the theoretical complexity of the full set of random polypeptides of a given length.","Such phage display libraries can be used to screen for amino acid combinations which will bind to a given target molecule.","Once a phage which binds to the desired target molecule is found, the peptide insert in that phage can be determined and molecules containing that peptide can be produced.","Such molecule would be expected to bind the target molecule.","The binding of the target molecule to its native receptor or ligand may be disrupted.","This is useful when it is desired to prevent the signaling which occurs when the target molecule is bound to its native ligand or receptor, all as is well known in the art of phage display libraries.","Furthermore, libraries can be prepared which incorporate the set of all of the overlapping oligonucleotides that represent all the overlapping peptides of a given antigen, or a large subset thereof, and, thus, create a phage display pepscan.","A scrambled pepscan, such as the combinatorial scrambled vaccines described in PCT application no.","WO 98/20169, the entire contents of which are hereby incorporated herein by reference, and the phage display two hybrid systems, such as are described in PCT application no.","WO 98/20159, the entire contents of which are hereby incorporated by reference, may also be prepared using the novel phages of the present invention.","Any of such phage display libraries can be used in screen assays as is known in the art.","In any such screening assay, the peptide that is found can then be produced and used for its intended purpose, again, as is well known in the prior art.","An example of such a screen would be to use a library in accordance with the present invention against a receptor.","In this way one might be able to select for a peptide that mimics the native ligand.","This peptide could then be produced using standard Merrifield synthesis, and the synthetic peptide may be used as either a lead peptide for further development or directly as a modulator of the receptor being studied.","For example, a random phage display peptide library may be used to screen against HIV gp120.In this way, one might be able to select for a peptide that binds to gp120 at precisely the CD4 binding site.","Such a peptide would be expected to bind to the virus and to prevent the association of the virus to the CD4 receptor and, thus, prevent infection.","One could also produce a library derived from CD4 itself and hope to discover a fragment of CD4 that binds to gp120 and acts as a decoy.","All possibilities stem from the fact that the library is produced in the genetically stable expression system of the present invention.","As to the selectable marker, a particular advantage of placing this marker between the two pVIII genes is to create a situation that if, for whatever reason, there is a genetic recombination based on the limited homology of the native pVIII gene and the recombinant pVIII gene, then the selectable marker would be excised and lost as well.","Running an experiment in the presence of the antibiotic would result in loss of those recombinants that have lost their resistance gene.","Other markers that could be used in place of the tetracycline resistance gene would be the ampicillin resistance gene, β-lactamase or chloramphenicol resistance gene.","As stated hereinabove, it is also possible within the scope of the present invention to also insert an exogenous gene between the pIII gene and its promoter.","By doing so, the phage can display a library of random peptides on either pIII or pVIII and a marker on the other, such as green fluorescence protein.","This would make the selection of the bound phage easier or amenable to high throughput screening.","The new exogenous gene can either be inserted directly between the pIII gene and its promoter or a second pIII gene can be inserted similarly to that discussed in the present invention for the second pVIII gene in order to ensure that the native pIII gene will always be produced along with the second pIII gene and the peptide of interest.","It is not necessary to insert a separate pIII gene as, in contrast to pVIII, pIII can tolerate large inserts.","The following examples are directed to preferred embodiments in accordance with the present invention and show how to make and use the present invention.","Those of ordinary skill in the art will understand, however, that these examples do not detract from the breadth of the present invention as described herein and that appropriate substitutions can be made without engaging in undue experimentation by those of ordinary skill in the art.","Materials and Methods Bacterial Strains, Phages, Reagents, and General Techniques E. coli strains used in this study were the following.","K802: F− e14−(McrA−) lacY1 or Δ(lac)6 supE44 galK2 galT22 rfbD1 metB1 mcrB1 hsdS3(rK− mK−).","K91KAN: a derivative of K91 (Hfr-Cavalli, thi) in which the “mini-Kan hopper” element was inserted in the lacZ gene of K91 rendering this strain kanamycin resistant (Parmley et al, 1988).","DH5α: F− endA1 hsdR17(rK− mK+) supE44 thi-1 recA1 gyrA(Nalr) relA1 Δ(lacIZYA-argF)U169 deoR (Φ80dlacΔ(lacZ)M15).","DH5αF′: F′ endA1 hsdR17(rK− mK+) supE44 thi-1 recA1 gyrA(Nalr) relA1 Δ(lacIZYA-argF)U169 deoR (Φ80dlacΔ(1acZ)M15).","JM109: F′[traD36 lacIq Δ(lacZ)M15 proA+B+] e14−(McrA−) Δ(lac-proAB) thi gyrA96(Nalr) endA1 hsdR17(rK− mK+) relA1 supE44 recA1.XL1-Blue: F′[::Tn10 proA+B+ lacIq Δ(lacZ)M15] recA1 endA1 gyrA96(Nalr) thi hsdR17(rK− mK+) supE44 relA1 lac.","MC1061: F− araD139 Δ(ara-leu)7696 galE15 galK16 Δ(lac)X74 rpsL(Strr) hsdR2(rK− mK+) mcrA mcrB1.NM554: F− araD139 Δ(ara-leu)7696 galE15 galK16 Δ(lac)X74 rpsL(Strr) hsdR2(rK− mK+) mcrA mcrB1 recA13.The wild type fd filamentous phage and fd-tet vector were kindly provided by G. P. Smith (University of Missouri, Columbia, Mo.).","The M13K07 phage was purchased from New England BioLabs Inc., MA (NEB).","DNA was isolated using the alkaline lysis procedure and purified on a cesium chloride gradient as described previously (Stern et al, 1998).","All the restriction enzymes used were purchased from NEB and the digestions were performed following the manufacturer's instructions.","The monoclonal antibody GV4H3 was produced from a Balb/c mouse immunized with the HIV-1 envelope protein gp120 (Denisova et al, 1996).","Oligonucleotides The oligonucleotides used in this study were the following.","ON1: 5′-GCCTTCGTAGTGGCATTACG-3′ (SEQ ID NO: 6) ON2: 5′-AGCCCGCTCATTAGGCGGGCTTCATTACCGGCCACGTCGGCCAC (SEQ ID NO: 7) CCTCAGCAGCGAAAGAC-3′ ON3: 5′-CGGTAATGAAGCCCGCCTAATGAGCGGGCTTTTTTTTGGCCTCT (SEQ ID NO: 8) GGGGCCGATCCCGCAAAAGCGGCC-3′ ON4: 5′-GGTCAGACGATTGGCCTTG-3′ (SEQ ID NO: 9) ON5: 5′-GAGCTCGCTAGCGCTCGAGCCAAAAAAAAAGGCTCCAAAAGG-3′ (SEQ ID NO: 10) ON6: 5′-CTAGAGCAGGGTCCAGCTAA-3′ (SEQ ID NO: 11) ON7: 5′-CTGGACCCTGCT-3′ (SEQ ID NO: 12) ON8: 5′-TCGAGCTGTTGACAATTAATCATCGGCTCGTATAATGTGTGGAA (SEQ ID NO: 13) TTGTGAGCGGATAACAATTGAGCT-3′ ON9: 5′-CAATTGTTATCCGCTCACAATTCCACACATTATACGAGCCGATG (SEQ ID NO: 14) ATTAATTGTCAACAGC-3′ ON10: 5′-CCGTGCATCTGTCCTCGTTC-3′ (SEQ ID NO: 15) ON11: 5′-CAGGCTTAAGCATCGACGTCTTATCAAGACGCCTTGCTTGTAA (SEQ ID NO: 16) ACTTTTTGAATAACTTGATACCGATAGTTGCGC-3′ ON12: 5′-GTCTTGATAAGACGTCGATGCTTAAGCCTGGCTAGCCATCAGAT (SEQ ID NO: 17) CTGAGTCGGCCGCTGTTTAAGAAATTCACCTCG-3′ ON13: 5′-GCCGTACCGCTAGCATTAGAAAAACTCATCGAGC-3′ (SEQ ID NO: 18) ON14: 5′-GCTTCCTGACAGGAGGCCGTTTTGTTTTGCAGCCCACCTGAGCT (SEQ ID NO: 19) CCCAGCTTAAGGTGTCTCAAAATCTCTGATG-3′ ON15: 5′-GGCTGAGGGACGTCGAGGGCATGCGTACCCGATAAAAGCGGCTT (SEQ ID NO: 20) CCTGACAGGAGGCCG-3′ ON16: 5′-AGCCTTTCAGGTCAGAAGGG-3′ (SEQ ID NO: 21) ON17: 5′-ATTCATGCCGGAGAGGGTAG-3′ (SEQ ID NO: 22) ON18: 5′-CATAACACCCCTTGTATTACTG-3′ (SEQ ID NO: 23) ON19: 5′-GCTTACATAAACAGTAATACAAGGGGTGTTATGAATAGTTCGAC (SEQ ID NO: 24) AAAGATCGC-3′ ON20: 5′-GGATCGAGAGCTAGCATAACTAAGCACTTGTCTCCTG-3′ (SEQ ID NO: 25) ON21: 5′-TGCCGGCGCTCCCGCTGGTTTCGCTATCCTGTCAGCACCCGGGT (SEQ ID NO: 26) CTG-3′ ON22: 5′-ACCCGGGTGCTGACAGGATAGCGAAACCAGCGGGAGCGCCGGCA (SEQ ID NO: 27) CGT-3′ Vector Construction In this study two novel vector-systems were constructed.","The rationale for their compositions and structures is described in the Results.","Detailed diagrams of the vectors and sub-structures are given in the Figures as indicated.","The following are the details of the specific steps taken to construct the vectors.","Construction of ftac88 The ftac88 vector (for detailed map see FIG.","7) was constructed through the following steps.","First, the recombinant pVIII gene, designated pVIIISTS (for orientation see FIG.","6A), was generated by introducing a 62 bp insert three codons downstream to the leader peptide using the “SOEing” PCR mutagenesis (Horton et al, 1990).","For this, four oligonucleotides were used.","ON1 and ON2 were used to PCR amplify from fd a 169 bp fragment.","ON3 and ON4 were used to PCR generate from fd a 899 bp fragment.","ON2 and ON3 each contain 5′-extension corresponding to the 62 bp insert.","Thus, the resulting PCR fragments contain an identical 30 bp stretch of the novel sequence.","The two fragments were purified from agarose gel, mixed and used as a combined template for PCR amplification using the oligonucleotides ON1 and ON4 to generate the final 1071 bp product containing the modified pVIII gene, part of pIII gene, and the flanking BamH I-SnaB I sites.","The PCR reaction was performed by using the Taq polymerase which adds adenine to the 3′ ends of PCR products and thus the resulting 1071 bp fragment was directly ligated with the linearized pGEM-T vector (Promega Corporation, WI) containing complementary 3′ thymidine overhangs, generating the pGEM-T(p8STS) construct (see FIG.","6B).","Next, the wild type pVIII gene was PCR amplified from fd using the oligonucleotides ON1 and ON5.ON5 has a 5′ extension that contains the restriction sites Xho I, Nhe I and Sac I.","The PCR reaction was performed using the Pwo Polymerase (Boehringer Mannheim), resulting in blunt-ended 335 bp fragment from the SnaB I site to just beyond the overlapping promoter/terminator followed by the ON5 introduced restriction sites.","The resulting fragment was ligated with the blunt-ended SnaB I linearized pGEM-T(p8STS) construct generating the pGEM-T(p88STS) construct (see FIG.","6C).","To remove the SnaB I site in tetR gene, the fd-tet vector (for detailed map see FIG.","5) was digested with Xba I and BstX I restriction enzymes and the 8458 bp fragment was purified from agarose gel.","The purified fragment was ligated with a linker containing the Ava II site produced by annealing the two complementary oligonucleotides ON6 and ON7.To exchange the SnaB I-BamH 1952 bp segment of fd-tet for the SnaB I-BamH 11378 bp segment of pGEM-T(p88STS) construct containing both the wild type and recombinant pVIII genes, the Ava II containing fd-tet derivative was digested with SnaB I/BamH I and the 7519 bp fragment was purified from agarose gel.","In the same manner, the pGEM-T(p88STS) construct was digested with SnaB I/BamH I and the released 1378 bp fragment was purified from agarose gel.","These two fragments were ligated to generate an intermediate vector.","This intermediate vector was digested with Sac I and Xho I restriction enzymes and the 13 bp stuffer was removed by applying the DNA digest on a chroma-spin™ column (Clontech Laboratories, Inc., Palo Alto, Calif.).","The linearized intermediate vector was ligated with a linker containing the tac promoter produced by annealing the two complementary oligonucleotides ON8 and ON9, generating the ftac88 vector.","Construction of fth1 The construction of the fth1 vector (for detailed map see FIG.","10) was performed through a series of intermediate vectors.","The intermediate vector-1 (IV-1, see FIG.","11B) was constructed by introducing a DNA segment ten codons upstream to the stop codon of the wild type pVIII gene using the “SOEing” PCR mutagenesis described above.","Two fragments of 519 bp and 827 bp were PCR generated from fd using the oligonucleotides ON10/ON11 and ON12/ON4 respectively.","The two fragments were mixed and used as a combined template using ON10/ON4 to PCR generate the final 1316 bp fragment.","This product was digested with SnaB I and BamH I and the resulting 1034 bp fragment was purified from agarose gel and used to replace the corresponding SnaB I-BamH I fragment of the Ava II containing fd-tet derivative described above for the construction of ftac88.Next, the IV-2 was constructed by introducing into the IV-1, downstream to the wild type pVIII gene, the novel HP transcription terminator followed by the kanamycin resistance gene.","This was performed in order (i) to functionally separate the two phage-transcriptional units and (ii) to enable the reconstitution of the intergenic region while maintaining an alternative antibiotic selectable marker (i.e.","kanamycin).","The kanamycin resistance gene was PCR amplified from M13K07 using the oligonucleotides ON13 and ON14.Both oligonucleotides contain 5′ extensions enabling the incorporation of additional flanking sequences.","The 3′ addition contained the Nhe I restriction site and the 5′ extension contained additional cloning sites adjacent to the kanamycin promoter and part of the HP terminator.","The resulting 944 bp fragment was purified from agarose gel and used as a template for PCR amplification using the oligonucleotides ON13 and ON15.The latter has a 5′ extension containing the second part of the HP terminator and additional cloning sites ending with the Aat II site.","Thus the resulting 984 bp fragment contains the entire HP terminator bracketed between multiple cloning sites followed by the kanamycin resistance gene.","This product was digested with Nhe I and Aat II and the resulting fragment was purified from agarose gel.","The purified fragment was ligated with Nhe I/Aat II digested IV-1 vector after removing the 20 bp stuffer by applying the vector digest on a chroma-spin™ column, ultimately generating IV-2.The IV-2 was used to reconstitute the intergenic region containing the origins of replication.","This was performed by PCR amplifying the intergenic region from fd phage using the oligonucleotides ON16 and ON17.The resulting 1081 bp fragment containing the Msc I-Drd I segment was digested with Msc I and Drd I restriction enzymes.","The 679 bp Msc I-Drd I product was purified from agarose gel and used to exchange the Msc I-Drd I fragment in IV-2 containing the tet fragment, generating IV-3.The open reading frame only of the kanamycin resistance gene was replaced by the open reading frame of the tetracycline resistance gene thus constructing the IV-4 vector (see FIG.","11C).","This was performed using the “SOEing” PCR.","The oligonucleotides ON1 and ON18 were used to PCR amplify from IV-2 the 463 bp fragment containing the upstream sequence to the ATG initiating codon of the kanamycin gene.","The oligonucleotides ON19 and ON20 were used to PCR amplify from fd-tet the 1265 bp fragment containing the open reading frame of the tetracycline resistance gene.","The two fragments were mixed and used for PCR amplification of the 1698 bp fragment using the oligonucleotides ON1 and ON20.The PCR was designed to flank the resulting 1698 bp fragment between Nhe I and Sac I restriction sites enabling the digestion of this fragment with the corresponding enzymes.","After purifying the digested fragment from agarose gel, it was used to replace the corresponding Nhe I-Sac I fragment of IV-3.The final step was to generate the synthetic recombinant pVIII gene using a panel of overlapping oligonucleotides (not shown).","Part of the panel was used as templates and the other part as 5′-extended primers for PCR amplification thus generating separately small pieces of DNA.","These small fragments were sequentially mixed and used as a combined template for the “SOEing” PCR ultimately generating the recombinant pVIIISTSh gene that subsequently was introduced between the Bgl II/Eag I sites of IV-4 producing the fth1 vector.","Introduction of the GV4H3 Epitope into ftac88 and fth1 Vectors The vectors ftac88 and fth1 were digested with Sfi I restriction enzyme and the 49 bp stuffer containing the trpA terminator (for orientation see FIG.","6a) was removed by applying the DNA digest on a chroma-spin™ column.","The purified linear vector was ligated with GV4H3 epitope encoding linker produced by annealing the two complementary oligonucleotides ON21 and ON22.Dot Blot Analysis of GV4H3 Epitope Presenting Phages The phages were applied via a vacuum manifold to a nitrocellulose membrane filter.","After blocking (5% evaporated spray dried skim milk 1.5% fat in Tris buffered saline (TBS)) for 1 hour, the membrane was washed briefly with TBS and incubated overnight with 1 μg/ml of GV4H3 mAb in TBS/5% milk at 4° C. with gentle rocking.","After washing, the membrane was incubated with goat-anti mouse IgG/HRP conjugate diluted 1:5000 in TBS/5% milk for 1 hour at room temperature.","The positive signals were detected by ECL (Amersham International plc, Buckinghamshire, England) immunodetection.","Results The fd-tet vector of Smith (Zacher et al, 1980) was used as a starting point for our studies.","The fd-tet Expression Vector Using the fd genome as a scaffold molecule, Zacher et al (1980) constructed the fd-tet phage, which contains the tetracycline resistance gene as a selectable marker (see FIG.","1).","The fragment coding for the tetracycline resistance was obtained from the transposon Tn10 (a 2775 bp fragment flanked between two Bgl II sites).","This segment was introduced into the BamH I site of the intergenic region containing the origins of replication of the fd genome.","The fd-tet vector has the following advantages: 1.fd-tet phages confer selectable tetracycline resistance to infected or transfected host bacteria.","2.The tet fragment introduces unique restriction sites that can be exploited for cloning foreign DNA sequences.","Thus, fd-tet was used, for example, as a cloning vector by exploiting the unique Hind III site, situated in the tet fragment, to clone Hind III digested phage X DNA (Zacher et al, 1980).","Furthermore, Smith has reported the introduction of a second pVIII gene in the same region generating a “type 88” vector designated f88.4, and the successful production of pVIII mosaic phages (Zhong et al, 1994).","However, a point for concern is the report that foreign DNA, cloned into the intergenic region of filamentous phages, tends to be unstable (Model et al, 1988; Sambrook et al, 1989; Vieira et al, 1987).","Occasionally, parts of the cloned DNA are deleted and the rate of the deletion event tends to be directly correlated with the size of the inserted fragment.","Therefore, the question of the genetic stability of DNA inserts cloned into the intergenic region was examined.","Genetic Instability in fd-tet Vector Purified fd-tet dsDNA generated two distinct electrophoretic patterns on agarose gels when isolated from two different E. coli strains (K802 and K91KAN).","The DNA isolated from K802 exhibited the supercoiled and relaxed forms of the fd-tet genome (FIG.","2A, lane 1) whereas the DNA isolated from K91KAN contained the supercoiled and relaxed forms of two distinct DNAs (FIG.","2A, lane 2).","One DNA corresponded to the fd-tet genome while the second compared well with the parent fd genome (FIG.","2A, lane 3).","The two purified fd-tet preparations and fd were then digested with the Msc I restriction enzyme that should generate full length linear DNA for both fd and fd-tet (the Msc I restriction site is unique in both fd and fd-tet, see FIG.","1).","Digestion of fd-tet from K802 and fd each generated only one band with the expected linear sizes of 9183 bp and 6414 bp respectively (FIG.","2B, lanes 1 and 3).","However, digestion of fd-tet derived from K91KAN generated two bands, one indistinguishable in mobility from the linear fd-tet and the second corresponding in size to linear fd (FIG.","2B, lane 2).","This indicates that for a substantial amount of DNA, a deletion of approximately 2.8 kb occurred when the fd-tet genome was produced in K91KAN bacteria.","Two points, therefore, must be addressed: 1.What DNA sequences are lost through the deletion process?","2.How are the differences in the bacterial strains related to the apparent genetic instability?","Deletion of the tet Fragment As the deleted DNA was 2.8 kbp, it was postulated that it might correspond to the tet-fragment (2775 bp) residing in the intergenic space of fd-tet (see FIG.","1).","Therefore, the SnaB I/Ava I digests of fd-tet DNA derived from both bacterial strains as before were compared.","The SnaB I site is unique in fd, however, it appears twice in fd-tet while the Ava I site exists only in the tet fragment of fd-tet.","The SnaB I/Ava I double digestion of fd-tet isolated from K802 generated three fragments with the expected sizes: 4381 bp, 2690 bp and 2112 bp (FIG.","3, lane 1).","The double digestion of the K91KAN derived fd-tet DNA, however, generated four DNA fragments (FIG.","3, lane 2); the three fragments corresponding to fd-tet and an additional fragment corresponding to the deletion product similar in size to a single cut full length 6414 bp fd (FIG.","3, lane 3, compare also with FIG.","2B).","This indicates that the SnaB I-Ava I fragment of the tet insert in the intergenic region was deleted from fd-tet.","This conclusion was further supported by BspH I/Hind III digestion of fd-tet derived from both bacteria.","The Hind III site is unique to fd-tet (see FIG.","1).","The BspH I site appears only once in fd and twice in fd-tet (see FIG.","1, the second BspH I site is situated in the tet fragment).","The BspH I/Hind III digestion of fd-tet DNA isolated from K802 generated the expected three fragments of 4676 bp, 2366 bp, and 2141 bp (FIG.","3, lane 4).","However, the BspH I/Hind III double digestion of fd-tet DNA isolated from K91KAN generated not only the three fd-tet derived fragments but also a fourth fragment (approximately 6.4 kb) corresponding in size to single cut full-length fd (FIG.","3, lane 5, note arrow) derived from the deletion product.","This not only confirms the selective deletion of the tet fragment but extends the boundaries of the deletion beyond the SnaB I site to at least the BspH I site (see FIG.","1).","Is the deletion a precise reversion to fd?","If so, one would reconstitute the original BamH I site used to clone the tet fragment, and thus generate two restriction fragments (3425 bp and 2989 bp as in the fd BamH I digest, see FIG.","4A lane 3) at the expense of the linearized deletion product (see for example the Msc I digest in FIG.","2B lane 2).","As can be seen in FIG.","4A (lane 2), BamH I digestion of fd-tet isolated from K91KAN produced a pattern similar to that shown for Msc I digest.","Two bands corresponding to the linearized fd-tet (9183 bp) and the full-length linearized deletion product (approximately 6.4 kb) were generated.","This thus illustrates that although the deletion encompasses much or all of the tet fragment, it does not precisely regenerate the second BamH I site of the fd parent molecule.","Thus, the next question addressed, once the 2.8 kbp deletion was mapped to the vicinity of the tet fragment, was whether or not the excision sites lay within or outside of the tet fragment.","For this the BstY I digest illustrated in FIG.","4B was performed (two BstY I sites, created during the fd-tet construction, precisely bracket the tet fragment, see FIG.","1).","The gel pattern obtained for the BstY I digest of fd-tet derived from K802 had three fragments (3425 bp, 2983 bp and 2775 bp) identical in size to those generated when K91KAN derived DNA was used (FIG.","4B, compare lane 1 with lane 2).","This demonstrates that at least one of the two BstY I sites was retained in the deleted product.","Therefore, sequences immediately adjacent to the original BamH I site, must also be preserved in the deleted fd-tet, a fact that may be relevant to the mechanism of the deletion process (see Discussion).","The Deletion is recA Independent and F+Dependent K802 and K91KAN strains of E. coli were introduced by Smith to produce and use the phage display vectors he has pioneered (Parmley, 1988).","The principle consideration is that phage infectability is dependent on the F-factor.","Therefore, manipulation of the DNA vector, its modifications and construction can be made in the absence of infection in F-bacteria (such as K802), thus avoiding multiply infected bacteria.","Once the vector is completed, then in order to screen a phage library or to amplify and generate high titers of phages one must use F+ bacteria (e.g., K91KAN).","Table 1 illustrates that in a variety of E. coli strains tested, the tet deletions were detected only in the F+ strains.","Most noteworthy is the comparison between the isogenic DH5α and DH5α F′ E. coli strains.","Here too the deletion occurs exclusively in the DH5αF′ strain.","These strains are identical in their genotypes except that DH5αF′ carries the F-factor.","This indicates that the deletion is F-factor dependent.","Moreover, both strains are RecA− in which the recA gene is mutated and thus classical homologous recombination is not possible.","Therefore, the deletion event is recA independent.","TABLE 1 The Deletion is F+ Dependent and recA Independent E. coli Strain redA F-episome Deletion K802 + − − K91KAN + + + DH5αa − − − DH5αF′a − + + JM109 − + + XL1-Blue − + + MC1061b + − − NM554b − − − aDH5α and DH5αF′ are isogenic strains; DH5αF′ contains the F-episome bMC1061 and NM554 are isogenic strains; NM554 is recA− As the F-factor is essential when using filamentous phages, working with F+ bacterial strains cannot be avoided.","Therefore, introducing the recombinant pVIII gene in a region prone to F+ dependent deletions is potentially problematic.","The suitability of the non-coding region between the wild type pVIII gene and the pIII gene as a site for genetic manipulation and introduction of a second pVIII gene was therefore explored.","Construction of the ftac88 Vector Designing a phage display vector with the aim of introducing a second pVIII gene in the non-coding region between the wild type pVIII and pIII genes presents two obstacles that must be considered.","1.Unique restriction sites for cloning are unavailable in this region.","2.This region contains an essential overlapping promoter and rho-independent transcription terminator that are inseparable (see FIG.","5; see also Van Wezenbeek et al, 1980).","The terminator terminates the transcription of upstream genes ending with the pVIII gene, whereas the promoter initiates the transcription of downstream genes starting with the pIII gene.","Therefore, the strategy adopted to insert the recombinant second pVIII gene was to duplicate this promoter/terminator as well.","Ultimately, a “type 88” expression vector (designated ftac88) was constructed as follows: The first step was to generate the modified pVIII gene.","This was designed to contain unique restriction sites that could be used to introduce foreign DNA sequences and thus enable the presentation of their corresponding encoded peptides at the N-terminus of the pVIII protein.","Such a modified pVIII gene, designated pVIIISTS, was produced by “SOEing” PCR (Horton et al, 1990) mutagenesis in which the GAC codon for residue Asp, at position 4 of mature wild type pVIII protein, was replaced by an insert of 62 base pairs (“STS” insert, FIG.","6A).","This insert contained two Sfi I sites that bracketed two translation stop-codons followed by the trpA transcription terminator.","Thus, the transcription of the pVIIISTS gene terminates prematurely, rendering the expression of this gene silent.","In order to express this pVIII gene one must exchange the Sfi I flanked stuffer for a functional in-frame insert of DNA.","The PCR was designed to generate a fragment corresponding to the SnaB I-BamH I segment of fd-tet (see FIG.","5) that was cloned into the pGEM-T vector, designated pGEM-T(p8STS) (FIG.","6B).","Next, a wild type pVIII gene was cloned into the SnaB I site of pGEM-T(p8STS).","This was achieved by PCR of the wild type pVIII gene segment from fd-tet starting from the upstream SnaB I site and continuing just beyond the overlapping promoter/terminator.","The downstream antisense primer incorporated three unique restriction sites as well.","The PCR product was cloned into the blunt ends of SnaB I cut pGEM-T(p8STS) thus generating the desired pGEM-T(p88STS).","As is illustrated in FIG.","6C the SnaB I-BamH I fragment of pGEM-T(p88STS) contains two pVIII genes, duplicated promoter/terminator elements and several unique cloning sites.","To complete the generation of the “type 88” phage display vector all that was required was to replace the SnaB I-BamH I segment of fd-tet with the corresponding SnaB I-BamH I fragment of pGEM-T(p88STS).","However, fd-tet contains two SnaB I sites (see FIG.","5).","Therefore, the SnaB I site in the tetR gene of fd-tet was removed.","For this, fd-tet genome was digested with Xba I/BstX I to remove the 725 bp fragment containing the SnaB I site that was replaced by a linker containing an Ava II site.","This intermediate modified fd-tet was then digested with SnaB I/BamH I and the wild type segment was exchanged for the correspondingly cut fragment derived from pGEM-T(p88STS).","Furthermore, a tac promoter was introduced between the Xho I and Sac I sites to produce the “type 88” vector designated ftac88 shown in FIG.","7.It was then necessary to evaluate whether or not the ftac88 vector can produce mosaic phages displaying both wild type and chimeric pVIII proteins.","For this a linker encoding the linear peptide APAGFAIL (SEQ ID NO:28) (the epitope corresponding to the anti-gp120 mAb GV4H3 (Denisova et al, 1996)) was inserted in frame between the two Sfi I sites, eliminating the trpA terminator.","As demonstrated by the dot blot analysis (FIG.","8), this construct (ftac88(4H3)) produced phages that were recognized by the GV4H3 mAb.","Genetic Instability in ftac88 The orientation of the wild type pVIII and the modified pVIIISTS genes in the ftac88 vector is a direct tandem repeat.","The construction of this vector was performed in recA− bacteria and as such excludes the possibility of recA dependent homologous recombination.","However, in view of the fact that the deletion of the tet fragment in fd-tet (described above) is recA independent and F+ dependent, the genetic stability of the modified pVIIISTS gene in DH5αF′ was examined.","For this, ftac88 DNA preparations from DH5α and DH5αF′ bacteria were compared.","Double digestion of ftac88 with SnaB I and BamH I should release a 1378 bp fragment containing both pVIII genes (see FIG.","7).","Indeed, SnaB I/BamH I double digestion produced only the expected 1378 bp fragment, regardless of the source of the DNA (FIG.","9, lane 1, 2).","As expected, the deletion of the tet fragment still occurred in DH5αF′ (FIG.","9, lane 2; indicated by the upper arrow).","Thus one can conclude that the modified pVIIISTS gene is genetically stable in ftac88.However, the purpose of the ftac88 vector is to display peptides as chimeric pVIII proteins and as such, it was important to further investigate the genetic stability of the modified pVIIISTS gene when it contained a peptide coding sequence rather than the stuffer with the trpA terminator.","For this, the ftac88 DNA construct expressing the GV4H3 epitope (above) was prepared from DH5α and DH5αF′ bacteria and used for the double digestion, SnaB I/BamH I analysis, as above.","Here the situation was surprising.","Digestion of the DNA derived from DH5α released as expected a 1376 bp fragment (FIG.","9, lane 3).","However, SnaB I/BamH I digestion of the same DNA prepared from DH5αF′ generated an additional smaller fragment of approximately 950 bp (FIG.","9, lane 4).","This indicates that a deletion of approximately 400 bp occurred.","This deletion corresponded to selective loss of the recombinant pVIII gene and reconstitution of the wild type pVIII-pIII configuration as confirmed by sequence analysis (not shown).","Therefore, it was concluded that the modified pVIIISTS gene, when containing coding sequences, becomes genetically unstable in DH5αF′.","This also suggests that the stability of the ftac88 observed in DH5αF′ is rendered due to the trpA terminator which could imply that the deletion process may require transcribed regions.","In summary, it is clear that the non-coding region between pVIII and pIII can be used.","Moreover, the obstacle of the overlapping promoter/terminator can be overcome.","However, genetic instability of the recombinant pVIII gene was still a problem.","Construction of fth1 Vector In light of the above experience in designing, constructing and testing the ftac88 vector, it was decided to redesign a novel “type 88” vector that would overcome the problems of genetic instability and include other improvements.","The attributes of such a vector, designated fth1 (FIG.","10) are listed herewith: 3.The second pVIII gene contained the 62 bp “STS” insert as in ftac88.However, the gene was constructed using synthetic oligonucleotides to generate a pVIII protein of identical amino acid sequence yet coded for with alternative codon usage.","This obviated the formation of tandem repeats, a configuration that is prone to deletion events as illustrated in the analysis of ftac88 (above).","The homologous gene was designated pVIIISTSh.","The pVIIISTSh gene was cloned, as in ftac88, between the wild type pVIII and pIII genes 4.To avoid the generation of additional tandem repeats, an alternative HP terminator (Nohno et al, 1986) was introduced into the 3′ region of the wild type pVIII gene ten codons upstream to the pVIII stop codon.","Moreover, the severed 3′ region coding for the C-terminal domain of the wild type pVIII protein was reconstituted using alternative codon usage (so to reduce homology within the −35 box of the promoter of the pIII gene residing in this region (Van Wezenbeek et al, 1980)).","5.The tetracycline resistance gene was cloned between the two pVIII genes.","This was accomplished via PCR of the open reading frame of the tetracycline resistance gene only and placing it downstream to a kanamycin promoter (derived from M13K07 phage).","This was done so as to avoid using the bi-directional tetracycline promoter (see FIG.","5) which would otherwise generate an anti-sense transcript of the wild type pVIII gene were it used in this new position.","Also the exclusion of the tetracycline repressor is advantageous as the tetracycline resistance gene thus becomes constitutively expressed, making pre-incubation with low levels of tetracycline to induce resistance unnecessary.","6.The original intergenic space containing the ori sequences was reconstituted.","Previously, the tet fragment used by Smith to construct fd-tet, was inserted into the fd ori(−) rendering this vector and its derivatives low copy number and thus producing relatively low phage titers (Smith, 1988).","Reconstituting the intergenic region in fth1 provided two advantages: the copy number of the DNA was greatly increased and the titers of secreted phage increased as well (see below).","The construction of fth1 (see FIG.","11) was accomplished using “SOEing” PCR as described above enabling the introduction of multiple cloning sites and novel genes into the non-coding region between the wild type pVIII and pIII genes of fd-tet.","The first step was to modify fd-tet by the introduction of a short segment of DNA, ten codons upstream from the stop codon of the wild type pVIII gene (FIG.","11B).","This segment not only provided novel restriction sites but also separated the promoter/terminator adjacent to the pIII gene from the wild type pVIII gene by reconstituting the severed last ten codons of the pVIII open reading frame (intermediate vector 1 (IV-1) in FIG.","11B).","Subsequently, additional intermediate vectors were produced (IV-2 and IV-3, see Materials and Methods) which eventually led to IV-4 (FIG.","1C).","In the latter, the novel HP terminator was incorporated downstream to the reconstituted wild type pVIII gene followed by the tetracycline resistance open reading frame driven by the Kanamycin promoter.","Moreover, the ori(−) in the intergenic region was also reconstituted in IV-4.The final vector was produced by cloning the synthetic pVIIISTSh gene, driven by the tac promoter, into the Bgl II/Eag I digested IV-4 vector to give fth1 (see detailed map, FIG.","10).","The questions to be evaluated regarding the utility and advantages of fth1 therefore were: 1.Was the vector, and particularly the tetracycline resistance and recombinant pVIII genes, genetically stable?","2.Could the recombinant pVIII gene be used to stably express peptide inserts?","3.Was the fth1 a high copy number vector?","4.Could high titers of fth1 progeny be achieved?","In order to answer these questions, the GV4H3 epitope was introduced between the Sfi I sites in fth1 (as done previously for ftac88).","This construct, called fth1(4H3), produced mosaic phages presenting the GV4H3 epitope that were recognized by the GV4H3 mAb (FIG.","8).","The stability of the tetracycline resistance gene and the modified pVIIISTSh gene was tested using restriction analyses as before.","Digestion of fth1 and fth1(4H3) DNA, derived from DH5α or DH5αF′, with BstN I generated identical fragments as is illustrated in FIG.","12.BstN I sites appear twice in both fd (see FIG.","1) and fth1 (see FIG.","10).","BstN I digestion of fd generated the expected two fragments of 5456 bp and 952 bp sizes (FIG.","12, lane 1).","The latter contains the non-coding region between pVIII and pIII genes.","As this region in fth1 contains the tetracycline resistance and the recombinant pVIIISTSh genes, the 952 bp fragment was shifted to generate the 2777 bp and 2775 bp bands in BstN I digests of fth1 and fth1(4H3) respectively derived from both DH5α and DH5αF′ (lane 2-5).","Any deletions of either the tetracycline resistance gene or the recombinant pVIII gene or their parts would generate DNA fragments ranging in size from 952 bp to 2777 bp.","No such fragments could be detected illustrating that the fth1 vector is genetically stable.","The functionality of the reconstituted ori(−) is directly apparent.","In several independent purifications of fth1 DNA, 0.5-1 mg DNA/liter of bacteria was routinely obtained, as opposed to the typical yields of 50 μg DNA per liter of ftac88 harboring bacteria.","Similarly a hundred fold increase is typically seen in phage titers for which 1012 phages/ml is measured for fth1 as compared to 1010 phages/ml achieved for fd-tet and its derivatives.","Thus the fth1 vector provides a convenient and stable “type 88” phage display expression system.","Discussion The selective deletion of the tet fragment from the intergenic space has been critically evaluated as a first step towards producing a novel “type 88” phage display vector.","The present inventors have observed that in F+ bacterial strains a 2.8 kb fragment of DNA is excised from the region of the foreign tet insert, Smith originally introduced into the B-stem/loop of the ori(−) structure (Zacher et al, 1980).","The loss of the insert's Ava I site, which overlaps the original remnant BamH I site, illustrates that downstream aspects of the insert are not preserved in the deleted form.","The loss of the BspH I site and all four of the insert's Hinc II sites (not shown) argues that no more than 18 base pairs of the insert can be retained at the other extreme.","Preservation of at least one BstY I site, yet lack of reconstitution of the original BamH I site, further stresses the close but not precise correspondence of the deleted fragment with the original tet insert.","As the insert is flanked by the inverted repeats that create the B-stem/loop and that these repeats are retained in the deleted form of the phage, one might postulate that the mechanism generating the deletion involves homology and base paring.","Clearly, however, this can not be homologous recombination as that would require direct repeats of homologous stretches of over 50 bases and of course, a functional RecA protein in order to enable a deletion event (Clark, 1973; Matfield et al, 1985).","The deletion events in our experiments occurred in recA− bacterial strains.","An alternative mechanism, the “slippage mechanism” proposed by Lovett (Feschenko et al, 1998), also requires direct repeats and therefore can not explain the deletion.","To the best of our knowledge, no known mechanism can explain the deletion of the tet fragment as observed.","One might imagine that during the course of replication, the inverted repeats, flanking the tet fragment, align to form a B-stem/loop of sorts, greatly distorted by a grossly extended “bubble” (where the BamH I site had been), consisting of the 2.8 kb insert.","During replication, the polymerase might be able to “skip” from one side of the bubble to the other thus omitting the entire insert and generating the deletion.","Assuming that this is a very rare event one must still address the fact that in K91Kan cells over 50% of the DNA isolated was of the deleted form.","Moreover, why is the deletion F+ dependent?","The matter of copy number might be explained due to two selective advantages that are realized in the deleted form.","The mere fact that the genome is 2.8 kb shorter than fd-tet, should speed up the replication.","More important may be the fact that the deletion most probably partially reconstitutes the ori(−) structure.","The ramifications of a markedly more functional ori(−) would substantially enable more effective production of double stranded DNA.","Another possibility may be related to the packaging of phages for secretion.","The A-stem/loop structure, situated adjacent to the B-stem/loop, is the packaging-signal of the phage (Van Wezenbeek et al, 1980).","It stands to reason that the tet insert may sterically hinder the packaging of the recombinant phages, a burden that would be removed in the deleted form.","However, the major phenomenon encountered here is the fact that the deletion is detectable only in the F+ bacteria.","This might be due to the fact that even rare events can be markedly amplified when super infection of bacteria can take place.","Obviously, for such super infection, the bacteria must be compatible, a situation only satisfied in F+ bacteria.","Therefore, it is proposed that the deletion leads to shorter and more efficient replicating forms of the phage, albeit a rare event.","Subsequently, however, such deleted phages can be markedly amplified in F+ bacteria via super infection thus enabling accumulation of very substantial amounts of the deleted form.","However, the possibility cannot be excluded that unknown proteins encoded by the F-episome might contribute to the deletion events either by directly enhancing the process or indirectly inducing the expression of bacterial proteins participating in the process.","In view of the above considerations, it was concluded that the intergenic region of the phage is not the optimal site for introduction of a second pVIII gene.","Not that this has not led to an effective “type 88” vector in the past such as the f88.4 vector constructed by Smith (Zhong et al, 1994).","In the f88.4 vector, the recombinant pVIII gene was introduced into the tet fragment of fd-tet.","However, one would expect with such a vector the gradual appearance of deleted forms of the phages and loss of their peptide inserts displayed by the chimeric pVIII protein.","Indeed, such “contaminating”, fast-growing, tetracycline sensitive phages have been reported by Scott when they used the f88.4 vector (Bonnycastle et al, 1996).","An alternative cloning site for the second pVIII gene was tested, namely the non-coding region between the pVIII and pIII genes.","The ftac88 described in this report illustrated that such a construction is not only possible but able to produce workable amounts of recombinant phages.","However, once again an unexpected obstacle was encountered as substantial amounts of deletions of the recombinant pVIII were observed.","This deletion process also occurred in a recA− background, excluding therefore the possibility of classical homologous recombination.","In contrast to the situation of the deletion of the tet insert however, the deletion of the modified pVIII gene entails reasonably long direct repeats.","Thus, whereas homologous recombination is not an option, the slippage mechanism which is recA independent, described by Feschenko and Lovett (Feschenko et al, 1998), might be responsible for the selective loss of the second pVIII gene in ftac88.This would entail the slipped misalignment of the nascent strand during DNA replication.","During replication of the repeated DNA the nascent strand becomes displaced from its template and pairs with the other copy on the template strand.","Then, replication is resumed and the slipped misalignment can lead to either deletion or expansion within tandem repeat arrays.","Therefore, deletions due to the slippage mechanism are homology dependent, a requirement satisfied in ftac88.The selective loss of the recombinant pVIII gene further supports the slippage mechanism.","This mechanism requires the stabilization of the slipped misalignment structure (Feschenko et al, 1998; Lovett et al, 1996).","The longer the displaced nascent strand, the more stabilized the slipped misalignment structure becomes.","The recombinant pVIII gene contains a foreign sequence in its 5′ region.","This severs the repeat into two parts, a short sequence upstream and a long downstream sequence.","The long sequence, therefore, enables the stabilization of the slipped misaligned structure more efficiently than the short one.","Therefore, excision sites in the long downstream identical sequence should predominate thus eliminating upstream sequences including the foreign DNA insert.","The actual deletion event may be a very rare one, typical of the slippage mechanism.","One can postulate that deleted DNA products could be amplified due to F+ dependent superinfection (see discussion above).","Therefore, the last aspect that must be considered is why the pVIIISTS form is stable as opposed to those constructs in which the stuffer containing the trpA terminator is exchanged for a peptide coding-sequence (such as the GV4H3 epitope).","The stability rendered by the presence of the trpA transcriptional terminator indicates that transcription might be a positive and necessary factor.","The trpA terminator stops the transcription of the recombinant pVIII gene prematurely and consequently no deletion is observed.","Transcription through the second homologous DNA repeat appears to be a necessary pre-requisite that provides the opportunity to form the misaligned structure that leads to subsequent deletion during DNA replication.","Such a mechanism may explain how misalignment can exist between the first and second repeats at the moment when the first repeat is being replicated yet the second downstream repeat is still in the closed double stranded DNA configuration.","In order to mispair the nascent strand to the closed double-stranded repeat, the latter would have to be opened.","In the absence of RecA protein, which catalyzes such reactions (West, 1992), transcription might be an efficient mode for opening double stranded DNA.","However, as opening double stranded DNA during transcription is transient, this mode would be efficient only when the DNA repeats are in reasonably close proximity.","Indeed, the deletion frequency, reported for the slippage mechanism, is correlated directly with the distance separating the DNA repeats (Lovett et al, 1994; Bi et al, 1994).","Moreover, this mechanism would require that the wild type pVIII gene be the first to be replicated.","Indeed, the most extensive replication during infection is that of the + strand, proceeding from the intergenic region through the wild type pVIII gene to the recombinant pVIII gene (Horiuchi et al, 1976).","As a result of the above analyses and the experiences gained through the construction and characterization of the ftac88 vector, an improved novel vector that corrects for the problems encountered was ultimately designed.","This vector has been designated fth1 and has the following advantages.","1.The tetracycline resistance and the recombinant pVIII genes were introduced between the wild type pVIII and pIII genes.","2.To avoid the presence of repetitive sequences of any kind, the two transcription units of the phage genes were separated by inserting a novel HP-terminator.","Furthermore, the wild type pVIII C-terminal domain encoding region was reconstituted using alternative codon usage.","The recombinant pVIII gene was generated using alternative codon usage as well.","These attributes render the fth1 vector genetically stable.","3.The fth1 vector contains the wild type intergenic region.","Thus, the minus-strand DNA synthesis occurs as efficiently as in wild type fd rendering fth1 a high copy number vector.","This enables one to isolate and purify consistently large amounts of the double-stranded DNA from fth1 harboring bacteria (500 μg per liter as oppose to 50 μg of ftac88).","Furthermore, fth1 produces high phage titers −1012 phages per ml as oppose to 1010 phages per ml typical for fd-tet and its derivatives.","The foregoing description of the specific embodiments will so fully reveal the general nature of the invention that others can, by applying current knowledge, readily modify and/or adapt for various applications such specific embodiments without undue experimentation and without departing from the generic concept, and, therefore, such adaptations and modifications should and are intended to be comprehended within the meaning and range of equivalents of the disclosed embodiments.","It is to be understood that the phraseology or terminology employed herein is for the purpose of description and not of limitation.","The means, materials, and steps for carrying out various disclosed functions may take a variety of alternative forms without departing from the invention.","Thus the expressions “means to .",".",".","” and “means for .",".",".","”, or any method step language, as may be found in the specification above and/or in the claims below, followed by a functional statement, are intended to define and cover whatever structural, physical, chemical or electrical element or structure, or whatever method step, which may now or in the future exist which carries out the recited function, whether or not precisely equivalent to the embodiment or embodiments disclosed in the specification above, i.e., other means or steps for carrying out the same functions can be used; and it is intended that such expressions be given their broadest interpretation.","REFERENCES Bi et al, “recA-independent and recA-dependent intramolecular plasmid recombination”, J Mol Biol 235:414-423 (1994) Bluthner et al, “Mapping of epitopes recognized by PM/Sc1 autoantibodies with gene-fragment phage display libraries”, J Immunol Methods 198:187-198 (1996) Bonnycastle et al, “Probing the basis of antibody reactivity with a panel of constrained peptide libraries displayed by filamentous phage”, J Mol Biol 258:747-762 (1996) Clark A J, “Recombination deficient mutants of Escherichia coli and other bacteria”, Ann Rev Genet 7:67-86 (1973) Cortese et al, “Epitope discovery using peptide libraries displayed on phage”, Trends Biotechnol 12:262-267 (1994) Cortese et al, “Identification of biologically active peptides using random libraries displayed on phage”, Curr Opin Biotechnol 6:73-80 (1995) Cortese et al, “Selection of biologically active peptides by phage display of random peptide libraries.","Curr Opin Biotechnol 7:616-621 (1996) Cwirla et al, “Peptides on phage: a vast library of peptides for identifying ligands”, Proc Natl Acad Sci USA 87:6378-6382 (1990) Denisova et al, “Humoral immune response to immunocomplexed HIV envelop glycoprotein 120”, AIDS Res Hum Retroviruses 12:901-909 (1996) Dotto et al, “The functional origin of bacteriophage fl DNA replication: Its Signal and domains”, J Mol Biol 172:507-521 (1984) Dower W J, “Phage power”, Curr Biol 2:251-253 (1992) Felici et al, “Selection of antibody ligands from a large library of oligopeptides expressed on a multivalent exposition vector”, J Mol Biol 222:301-310 (1991) Feschenko et al, “Slipped misalignment mechanisms of deletion formation: analysis of deletion endpoints”, J Mol Biol 276:559-569 (1998) Goldsmith et al, “Adsorption protein of bacteriophage fd: Isolation, molecular properties, and location in virus”, Biochemistry 16:2686-2694 (1977) Gray et al “Adsorption complex of filamentous fd virus”, J Mol Biol 146:621-627 (1981) Greenwood et al, “Multiple display of foreign peptides on a filamentous bacteriophage”, J Mol Biol 220:821-827 (1991) Hoess et al, “Identification of a peptide which binds to the carbohydrate-specific monoclonal antibody B3”, Gene 128:43-49 (1993) Holliger et al, “A conserved infection pathway for filamentous bacteriophages is suggested by the structure of the membrane penetration domain of the minor coat protein g3p from phage fd”, Structure 5:265-275 (1997) Horiuchi et al, “Origin and direction of synthesis of bacteriophage f1 DNA”, Proc Natl Acad Sci USA 73:2341-2345 (1976) Horton et al, “Gene splicing by overlap extension: tailor-made genes using the polymerase chain reaction”, Biotechniques 8:528-535 (1990) Iannolo et al, “Modifying filamentous phage capsid: limits in the size of the major capsid protein”, J Mol Biol 248:835-844 (1995) Kay et al, “An M13 phage library displaying 38-amino-acid peptides as a source of novel sequences with affinity to selected targets”, Gene 128:59-65 (1993) Kim et al, “Viable deletions of the M13 complementary strand origin”, Proc Natl Acad Sci USA 78:6784-6788 (1981) Koivunen et al, “Phage libraries displaying cyclic peptides with different ring sizes: ligand specificities of the RGD-directed integrins”, Biotechnology 13:265-270 (1995) Krebber et al, “Co-selection of cognate-antigen pairs by selectively-infective phages”, FEBS Lett 377:227-231 (1995) Lane et al, “Epitope mapping using bacteriophage peptide libraries”, Curr Opin Immunol 5:268-271 (1993) Lovett et al, “Recombination between repeats in Escherichia coli by a recA-independent, proximity-sensitive mechanism”, Mol Gen Genet 245:294-300 (1994) Lovett et al, “Stabilization of diverged tandem repeats by mismatch repair: evidence for deletion formation via a misaligned replication intermediate”, Proc Natl Acad Sci USA 93:7120-7124 (1996) Luzzago et al, “Mimicking of discontinuous epitopes by phage displayed peptides, I. Epitope mapping of human H ferritin using a phage library of constrained peptides”, Gene 128:51-57 (1993) Marvin et al, “Molecular model and structural comparisons of native and mutant class I filamentous bacteriophages Ff (fd, f1, M13), If1 and IKe”, J Mol Biol 235:260-286 (1994) Matfield et al, “Rec-dependent and Rec-independent recombination of plasmid-borne duplications in Escherichia coli K12”, Mol Gen Genet 199:518-523 (1985) Matthews et al, “Substrate phage: selection of protease substrates by monovalent phage display”, Science 260:1113-1116 (1993) McCafferty et al, “Phage antibodies: filamentous phage displaying antibody variable domains”, Nature 348:552-554 (1990) McCafferty et al, “Phage enzymes: expression and affinity chromatography of functional alkaline phosphatase on the surface of bacteriophage”, Protein Eng 4:955-961 (1992) Messing J, “New M13 vectors for cloning”, Methods Enzymol 101:20-78 (1983) Model et al, “Filamentous Bacteriophage”, in The Bacteriophages, Calendar R (ed.","), Plenum Press, New York and London, Vol.","2, p. 375 (1988) Moses et al, “Restructuring the bacteriophage f1 genome: Expression of gene VIII in the intergenic space”, Virology 104:267-278 (1980) Nohno et al, “Cloning and complete sequence of the Escherichia coli glutamine permease operon (glnHPQ)”, Mol Gen Genet 205:260-269 (1986) Parmley et al, “Antibody-selectable filamentous fd phage vectors: affinity purification of target genes”, Gene 73:305-318 (1988) Rasqualini et al, “Organ targeting in vivo using phage display peptide libraries”, Nature 380:364-366 (1996) Sambrook et al, Molecular Cloning: a Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1989) Schmitz et al, “Catalytic specificity of phosphotyrosine kinases Blk, Lyn, c-Src and Syk as assessed by phage display”, J Mol Biol 260:664-677 (1996) Scott et al, “Searching for peptide ligands with an epitope library”, Science 249:386-390 (1990) Smith G P, “Filamentous phage assembly: morphogenetically defective mutants that do not kill the host”, Virology 167:156-165 (1988) Smith G P “Surface display and peptide libraries”, Gene 128:1-2 (1993) Sparks et al, “Identification and characterization of Src SH3 ligands from phage-displayed random peptide libraries”, J Biol Chem 269:23853-23856 (1994) Stern et al, “Construction and use of a 20-mer phage display epitope library”, in Methods in Molecular Biology: Combinatorial Peptide Library Protocols, S. Cabilly (ed.","), Humana Press, Totowa, N.J., p. 137 (1998) Sugimoto et al, “Studies on bacteriophage fd DNA.","IV.","The sequence of messenger RNA for the major coat protein gene”, J Mol Biol 111:487-507 (1977) Van Wezenbeek et al, “Nucleotide sequence of the filamentous bacteriophage M13 DNA genome: comparison with phage fd”, Gene 11:129-148 (1980) Vieira et al, “Production of single-stranded plasmid DNA”, Methods Enzymol 153:3-11 (1987) West S C, “Enzymes and molecular mechanisms of genetic recombination”, Annu Rev Biochem 61:603-640 (1992) Willis et al, “Immunological properties of foreign peptides in multiple display on a filamentous bacteriophage”, Gene 128:79-83 (1993) Wrighton et al, “Small peptides as potent mimetics of the protein hormone erythropoietin”, Science 273:458-463 (1996) Zacher et al, “A new filamentous phage cloning vector: fd-tet”, Gene 9:127-140 (1980) Zhong et al, “Conformational mimicry of a chlamydial neutralization epitope on filamentous phage”, J Biol Chem 269:24183-24188 (1994"]],"string":"[\n [\n \"Phage display vector, phages encoded thereby, and methods of use thereof\",\n \"Filamentous bacteriophages are particularly efficient for the expression and display of combinatorial random peptides.\",\n \"Two phage proteins are often employed for peptide display: the infectivity protein, pIII and the major coat protein, pVIII.\",\n \"The use of pVIII typically requires the expression of two pVIII genes: the wild type and the recombinant pVIII genes to generate mosaic phages.\",\n \"“Type 88” vectors contain two pVIII genes in one phage genome.\",\n \"A novel “type 88” expression vector has been rationally designed and constructed, which can be used to express recombinant peptides as pVIII chimeric proteins in mosaic bacteriophages.\",\n \"This vector is not only genetically stable but also of high copy number and produces high titers of recombinant phages.\"\n ],\n [\n \"1.A DNA molecule comprising the DNA of a filamentous phage into which has been inserted a recombinant DNA sequence comprising a multiple cloning site comprising a series of restriction sites that do not otherwise appear in said DNA of the filamentous phage, said recombinant DNA sequence being designed and inserted such that the multiple cloning site is placed between a terminator for the wild type pVIII gene and an initiator for the wild type pIII gene.\",\n \"2.A DNA molecule in accordance with claim 1, wherein a positive selection marker is present in the multiple cloning site.\",\n \"3.A DNA molecule in accordance with claim 1, wherein an additional bacteriophage gene is present in the multiple cloning site.\",\n \"4.A DNA molecule in accordance with claim 1, wherein said additional bacteriophage gene is a second pVIII gene.\",\n \"5.A DNA molecule in accordance with claim 1, wherein said additional bacteriophage gene is a second pIII gene.\",\n \"6.A DNA molecule in accordance with claim 1, wherein said recombinant DNA sequence is designed and inserted such that a DNA sequence encoding a foreign peptide is incorporated into the pIII gene at the N-terminal region thereof.\",\n \"7.A DNA molecule in accordance with claim 1, wherein said multiple cloning site placed between said wild type pVIII gene (the first pVIII gene) and the wild type pIII gene comprises DNA encoding an additional pVIII protein (the second pVIII gene) and DNA encoding a polypeptide of interest other than pVIII.\",\n \"8.A DNA molecule in accordance with claim 7, wherein the DNA encoding the second pVIII gene uses alternative codons which are other than the native codons for encoding one or more of the amino acid residues of the pVIII protein, such that no stretch of more than 30 bases of the native sequence appear therein.\",\n \"9.A DNA molecule in accordance with claim 8, wherein no stretch of more than 20 bases of the native sequence appear in the DNA encoding the second pVIII gene.\",\n \"10.A DNA molecule in accordance with claim 7, wherein the native pVIII DNA sequence is separated from the recombinant pVIII DNA sequence by a terminator which is not native to the filamentous phage DNA.\",\n \"11.A DNA molecule in accordance with claim 7, wherein DNA encoding the native amino acid sequence of the pVIII C-terminal/pIII −35 box is maintained at the region immediately upstream to the initiation of the pIII gene.\",\n \"12.A DNA molecule in accordance with claim 11, wherein said region encoding the pVIII C-terminal domain in said C-terminal/−35 box uses alternative codons which are other than the native codons, such that no stretch of more than 30 bases of the native sequence appear therein.\",\n \"13.A DNA molecule in accordance with claim 12, wherein no stretch of more than 20 bases of the native sequence appear in the region encoding the pVIII C-terminal/−35 box.\",\n \"14.A DNA molecule in accordance with claim 7, wherein a positive selection marker is inserted between the native and recombinant pVIII genes.\",\n \"15.A DNA molecule in accordance with claim 14, wherein said positive selection marker is the tetracycline resistance gene.\",\n \"16.A DNA molecule in accordance with claim 15, wherein a unidirectional promoter is substituted for the native bidirectional tetracycline resistance gene promoter.\",\n \"17.A DNA molecule in accordance with claim 7, wherein the native intergenic space region is present.\",\n \"18.A DNA molecule in accordance with claim 7, wherein a promoter is used for the second pVIII gene and DNA encoding the peptide of interest, which promoter is not native to the native filamentous phage.\",\n \"19.A DNA molecule in accordance with claim 7, wherein said recombinant DNA sequence is designed and inserted such that a DNA sequence encoding a second peptide of interest is incorporated into the pIII gene at the N-terminal region thereof.\",\n \"20.A filamentous phage encoded by the DNA of claim 7.21.A DNA molecule in accordance with claim 7 wherein said polypeptide of interest is a random peptide.\",\n \"22.A library of filamentous phages encoded by a DNA molecule in accordance with claim 21, wherein said library comprises a plurality of phages each having a different random peptide as the polypeptide of interest.\",\n \"23.A library in accordance with claim 22, wherein the polypeptides of interest in said plurality of phages comprise all possible random peptides of a given length or a fraction thereof which maintains the complexity thereof.\",\n \"24.A filamentous phage encoded by a DNA molecule in accordance with claim 7, wherein said polypeptide of interest is a peptide which comprises a contiguous fragment of the sequence of an antigen of interest.\",\n \"25.A library of filamentous phages in accordance with claim 24, wherein said library comprises a plurality of phages, said peptide in each of said plurality of phages comprising a different contiguous fragment of the sequence of the antigen of interest.\",\n \"26.A library in accordance with claim 25, wherein the polypeptides of interest in said plurality of phages comprise all possible overlapping contiguous fragments of the antigen of interest.\",\n \"27.A method of screening for a molecule which binds to a peptide of interest comprising bringing a molecule to be screened into contact with a filamentous phage in accordance with claim 20 which expresses said peptide of interest and, if said molecule binds to said peptide of interest, identifying and producing said molecule.\",\n \"28.A method of screening for peptides which bind to a molecule of interest comprising bringing the molecule of interest into contact with a library of filamentous phages in accordance with claim 22; identifying any peptide expressed by a phage which binds to said molecule; and producing a polypeptide which comprises said peptide.\",\n \"29.A method of screening for peptides which bind to a molecule of interest comprising bringing the molecule of interest into contact with a library of filamentous phages in accordance with claim 25; identifying any peptide expressed by a phage which binds to said molecule; and producing a polypeptide which comprises said peptide.\",\n \"30.A DNA molecule comprising the DNA of a filamentous phage, into which has been inserted between the wild type pVIII gene (the first pVIII gene) and the wild type pIII gene, recombinant DNA sequence encoding an additional pVIII protein (the second pVIII gene) and DNA containing a region of bases flanked by two identical restriction enzyme sites, which sites do not appear anywhere else in the DNA molecule.\",\n \"31.A DNA construct for insertion into the DNA of a filamentous phage, said construct comprising a multiple cloning site comprising a series of restriction sites that do not otherwise appear in the DNA of the phage into which the construct is intended to be inserted, said construct having one end compatible for ligation to a restriction enzyme site upstream of the pIII gene of the phage into which the construct is intended to be inserted and another end compatible for ligation to a restriction enzyme site downstream of the pVIII gene of the phage into which the construct is intended to be inserted.\",\n \"32.A DNA construct in accordance with claim 31 further including a positive selection marker.\",\n \"33.A DNA construct in accordance with claim 31 further including a second pVIII gene.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>Combinatorial phage display peptide libraries provide an effective means to study protein:protein interactions.\",\n \"This technology relies on the production of very large collections of random peptides associated with their corresponding genetic blueprints (Scott et al, 1990; Dower, 1992; Lane et al, 1993; Cortese et al, 1994; Cortese et al, 1995; Cortese et al, 1996).\",\n \"Presentation of the random peptides is often accomplished by constructing chimeric proteins expressed on the outer surface of filamentous bacteriophages such as M13, fd and f1.This presentation makes the repertoires amenable to binding assays and specialized screening schemes (referred to as biopanning (Parmley et al, 1988)) leading to the affinity isolation and identification of peptides with desired binding properties.\",\n \"In this way peptides that bind to receptors (Koivunen et al, 1995; Wrighton et al, 1996; Sparks et all, 1994; Rasqualini et al, 1996), enzymes (Matthews et al, 1993; Schmitz et al, 1996) or antibodies (Scott et al, 1990; Cwirla et al, 1990; Felici et al, 1991; Luzzago et al, 1993; Hoess et al, 1993; Bonnycastle et al, 1996) have been efficiently selected.\",\n \"Filamentous bacteriophages are nonlytic, male specific bacteriophages that infect Escherichia coli cells carrying an F-episome (for review, see Model et al, 1988).\",\n \"Filamentous phage particles appear as thin tubular structures 900 nm long and 10 nm thick containing a circular single stranded DNA genome (the +strand).\",\n \"The life cycle of the phage entails binding of the phage to the F-pilus of the bacterium followed by entry of the single stranded DNA genome into the host.\",\n \"The circular single stranded DNA is recognized by the host replication machinery and the synthesis of the complementary second DNA strand is initiated at the phage ori(−) structure.\",\n \"The double stranded DNA replicating form is the template for the synthesis of single-stranded DNA circular phage genomes, initiating at the ori(+) structure.\",\n \"These are ultimately packaged into virions and the phage particles are extruded from the bacterium without causing lysis or apparent damage to the host.\",\n \"Peptide display systems have exploited two structural proteins of the phage; pIII protein and pVIII protein.\",\n \"The pIII protein exists in 5 copies per phage and is found exclusively at one tip of the virion (Goldsmith et al, 1977).\",\n \"The N-terminal domain of the pIII protein forms a knob-like structure that is required for the infectivity process (Gray et al, 1981).\",\n \"It enables the adsorption of the phage to the tip of the F-pilus and subsequently the penetration and translocation of the single stranded phage DNA into the bacterial host cell (Holliger et al, 1997).\",\n \"The pIII protein can tolerate extensive modifications and thus has been used to express peptides at its N-terminus.\",\n \"The foreign peptides have been up to 65 amino acid residues long (Bluthner et al, 1996; Kay et al, 1993) and in some instances even as large as full-length proteins (McCafferty et al, 1990; McCafferty et al, 1992) without markedly affecting pIII function.\",\n \"The cylindrical protein envelope surrounding the single stranded phage DNA is composed of 2700 copies of the major coat protein, pVIII, an α-helical subunit which consists of 50 amino acid residues.\",\n \"The pVIII proteins themselves are arranged in a helical pattern, with the α-helix of the protein oriented at a shallow angle to the long axis of the virion (Marvin et al, 1994).\",\n \"The primary structure of this protein contains three separate domains: (1) the N-terminal part, enriched with acidic amino acids and exposed to the outside environment; (2) a central hydrophobic domain responsible for: (i) subunit:subunit interactions in the phage particle and (ii) transmembrane functions in the host cell; and (3) the third domain containing basic amino acids, clustered at the C-terminus, which is buried in the interior of the phage and is associated with the phage-DNA.\",\n \"pVIII is synthesized as a precoat protein containing a 23 amino acid leader-peptide, which is cleaved upon translocation across the inner membrane of the bacterium to yield the mature 50-residue transmembrane protein (Sugimoto et al, 1977).\",\n \"Use of pVIII as a display scaffold is hindered by the fact that it can tolerate the addition of peptides no longer than 6 residues at its N-terminus (Greenwood et al, 1991; Iannolo et al, 1995).\",\n \"Larger inserts interfere with phage assembly.\",\n \"Introduction of larger peptides, however, is possible in systems where mosaic phages are produced by in vivo mixing the recombinant, peptide-containing, pVIII proteins with wild type pVIII (Felici et al, 1991; Greenwood et al, 1991; Willis et al, 1993).\",\n \"This enables the incorporation of the chimeric pVIII proteins at low density (tens to hundreds of copies per particle) on the phage surface interspersed with wild type coat proteins during the assembly of phage particles.\",\n \"Two systems have been used that enable the generation of mosaic phages; the “type 8+8” and “type 88” systems as designated by Smith (Smith, 1993).\",\n \"The “type 8+8” system is based on having the two pVIII genes situated separately in two different genetic units (Felici et al, 1991; Greenwood et al, 1991; Willis et al, 1993).\",\n \"The recombinant pVIII gene is located on a phagemid, a plasmid that contains, in addition to its own origin of replication, the phage origins of replication and packaging signal.\",\n \"The wild type pVIII protein is supplied by superinfecting phagemid-harboring bacteria with a helper phage.\",\n \"In addition, the helper phage provides the phage replication and assembly machinery that package both the phagemid and the helper genomes into virions.\",\n \"Therefore, two types of particles are secreted by such bacteria, helper and phagemid, both of which incorporate a mixture of recombinant and wild type pVIII proteins.\",\n \"The “type 88” system benefits by containing the two pVIII genes in one and the same infectious phage genome.\",\n \"Thus, this obviates the need for a helper phage and superinfection.\",\n \"Furthermore, only one type of mosaic phage is produced.\",\n \"The question arises, however, where one should introduce the second pVIII gene within the filamentous phage genome for efficient expression and genetic stability.\",\n \"The phage genome encodes 10 proteins (pI through pX) all of which are essential for production of infectious progeny (Felici et al, 1991).\",\n \"The genes for the proteins are organized in two tightly packed transcriptional units separated by two non-coding regions (Van Wezenbeek et al, 1980).\",\n \"One non-coding region, called the “intergenic region” (defined as situated between the pIV and pII genes) contains the (+) and the (−) origins of DNA replication and the packaging signal of the phage, enabling the initiation of capsid formation.\",\n \"Parts of this intergenic region are dispensable (Kim et al, 1981; Dotto et al, 1984).\",\n \"Moreover, this region has been found to be able to tolerate the insertion of foreign DNAs at several sites (Messing, 1983; Moses et al, 1980; Zacher et al, 1980).\",\n \"The second non-coding region of the phage is located between the pVIII and pIII genes, and has also been used to incorporate foreign recombinant genes as was illustrated by Pluckthun (Krebber et al, 1995).\",\n \"Regardless as to where a second pVIII gene is to be introduced, a major point for concern is the genetic stability of the ultimate vector and its derivatives.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>The present invention is based on a critical examination of the attributes of the two non-coding regions of the fd filamentous phage as potential sites for insertion of a second recombinant pVIII gene and its genetic stability, resulting in the design and construction of an efficient “type 88” phage display expression system.\",\n \"The phage display expression system of the present invention includes a vector which is the DNA sequence of a filamentous phage into which a second pVIII gene, as well as DNA encoding a peptide of interest, are placed between the wild type pVIII and pIII genes.\",\n \"This allows production of type 88 phages displaying a peptide of interest with genetic stability and high copy number.\",\n \"The DNA encoding the second pVIII gene preferably uses alternative codons for encoding the amino acid residues of the pVIII protein.\",\n \"The native pVIII DNA sequence is separated from the recombinant pVIII sequence by a region encoding the wild type pVIII C-terminal domain and a terminator.\",\n \"The region encoding the wild type pVIII C-terminal domain is preferably designed to use alternative codons which are other than the native codons for encoding the same amino acids.\",\n \"In this way homologous recombination, slippage mechanisms and other genetic instabilities may be avoided.\",\n \"Preferably, the terminator at the end of the native pVIII DNA sequence is an HP terminator which is not native to the filamentous phage DNA.\",\n \"In a preferred embodiment of the present invention, a positive selection marker, such as the tetracycline or kanamycin resistant genes, is inserted between the native and recombinant pVIII genes.\",\n \"Preferably, a unidirectional promoter is substituted for the native bidirectional tetracycline resistant gene promoter.\",\n \"In preferred embodiments of the present invention, the native intergenic space between pIV and pII is maintained.\",\n \"The present invention further relates to the filamentous phages encoded by the DNA discussed above.\",\n \"Such phages may be of any type, such as fd, M13 and f1, although fd is preferred.\",\n \"The peptide designed to be displayed by the phage of the present invention can be any peptide which is desired to be presented, such as a specific antigen.\",\n \"Indeed, any protein or peptide can be displayed on the phage of the present invention.\",\n \"The peptide displayed on the phage may be an epitope of an antigen, which phage can be used therapeutically as a vaccine.\",\n \"Furthermore, the peptide displayed on the phage may be a single-chain antibody, which phage can be used for passive immunization.\",\n \"Both of these embodiments are described in WO 01/18169.Alternatively, one can create a library of phages into which are incorporated, at the same site, each of a set of oligonucleotides that encode all possible random peptides of a given length, or a large subset of that set.\",\n \"Such a large subset should preferably represent a fraction of the full set with the theoretical complexity of the full set of random polypeptides of a given length.\",\n \"Libraries of phages can also be prepared which incorporate all, or a large subset of all, of the overlapping oligonucleotides that represent all of the overlapping peptides of a given antigen and, thus, create a phage display pepscan.\",\n \"Scrambled pepscans (PCT application no.\",\n \"WO 98/20169) and phage display two hybrid systems (PCT application no.\",\n \"WO 98/20159) may also be prepared by this technique.\",\n \"The phage display libraries of the present invention can be used in screening for molecules which bind to a particular displayed peptide of interest or in screening the peptides displayed on a library of phages to see which bind to a specific molecule of interest, all as is well known with respect to prior art phage display libraries.\",\n \"Once a molecule or peptide of interest is identified by means of the screen using the phages or phage display libraries of the present invention, the peptide or molecule so identified may be produced in a conventional manner.\"\n ],\n [\n \"FIELD OF THE INVENTION The present invention is related to a phage display vector encoding a filamentous phage which includes DNA encoding a polypeptide of interest and, more particularly, to such a vector in which a recombinant sequence including a multiple cloning site is inserted between the wild type pVIII and pIII genes.\",\n \"The invention further relates to the phages encoded by such DNA, phage display libraries made of such phages and methods of use thereof.\",\n \"BACKGROUND OF THE INVENTION Combinatorial phage display peptide libraries provide an effective means to study protein:protein interactions.\",\n \"This technology relies on the production of very large collections of random peptides associated with their corresponding genetic blueprints (Scott et al, 1990; Dower, 1992; Lane et al, 1993; Cortese et al, 1994; Cortese et al, 1995; Cortese et al, 1996).\",\n \"Presentation of the random peptides is often accomplished by constructing chimeric proteins expressed on the outer surface of filamentous bacteriophages such as M13, fd and f1.This presentation makes the repertoires amenable to binding assays and specialized screening schemes (referred to as biopanning (Parmley et al, 1988)) leading to the affinity isolation and identification of peptides with desired binding properties.\",\n \"In this way peptides that bind to receptors (Koivunen et al, 1995; Wrighton et al, 1996; Sparks et all, 1994; Rasqualini et al, 1996), enzymes (Matthews et al, 1993; Schmitz et al, 1996) or antibodies (Scott et al, 1990; Cwirla et al, 1990; Felici et al, 1991; Luzzago et al, 1993; Hoess et al, 1993; Bonnycastle et al, 1996) have been efficiently selected.\",\n \"Filamentous bacteriophages are nonlytic, male specific bacteriophages that infect Escherichia coli cells carrying an F-episome (for review, see Model et al, 1988).\",\n \"Filamentous phage particles appear as thin tubular structures 900 nm long and 10 nm thick containing a circular single stranded DNA genome (the +strand).\",\n \"The life cycle of the phage entails binding of the phage to the F-pilus of the bacterium followed by entry of the single stranded DNA genome into the host.\",\n \"The circular single stranded DNA is recognized by the host replication machinery and the synthesis of the complementary second DNA strand is initiated at the phage ori(−) structure.\",\n \"The double stranded DNA replicating form is the template for the synthesis of single-stranded DNA circular phage genomes, initiating at the ori(+) structure.\",\n \"These are ultimately packaged into virions and the phage particles are extruded from the bacterium without causing lysis or apparent damage to the host.\",\n \"Peptide display systems have exploited two structural proteins of the phage; pIII protein and pVIII protein.\",\n \"The pIII protein exists in 5 copies per phage and is found exclusively at one tip of the virion (Goldsmith et al, 1977).\",\n \"The N-terminal domain of the pIII protein forms a knob-like structure that is required for the infectivity process (Gray et al, 1981).\",\n \"It enables the adsorption of the phage to the tip of the F-pilus and subsequently the penetration and translocation of the single stranded phage DNA into the bacterial host cell (Holliger et al, 1997).\",\n \"The pIII protein can tolerate extensive modifications and thus has been used to express peptides at its N-terminus.\",\n \"The foreign peptides have been up to 65 amino acid residues long (Bluthner et al, 1996; Kay et al, 1993) and in some instances even as large as full-length proteins (McCafferty et al, 1990; McCafferty et al, 1992) without markedly affecting pIII function.\",\n \"The cylindrical protein envelope surrounding the single stranded phage DNA is composed of 2700 copies of the major coat protein, pVIII, an α-helical subunit which consists of 50 amino acid residues.\",\n \"The pVIII proteins themselves are arranged in a helical pattern, with the α-helix of the protein oriented at a shallow angle to the long axis of the virion (Marvin et al, 1994).\",\n \"The primary structure of this protein contains three separate domains: (1) the N-terminal part, enriched with acidic amino acids and exposed to the outside environment; (2) a central hydrophobic domain responsible for: (i) subunit:subunit interactions in the phage particle and (ii) transmembrane functions in the host cell; and (3) the third domain containing basic amino acids, clustered at the C-terminus, which is buried in the interior of the phage and is associated with the phage-DNA.\",\n \"pVIII is synthesized as a precoat protein containing a 23 amino acid leader-peptide, which is cleaved upon translocation across the inner membrane of the bacterium to yield the mature 50-residue transmembrane protein (Sugimoto et al, 1977).\",\n \"Use of pVIII as a display scaffold is hindered by the fact that it can tolerate the addition of peptides no longer than 6 residues at its N-terminus (Greenwood et al, 1991; Iannolo et al, 1995).\",\n \"Larger inserts interfere with phage assembly.\",\n \"Introduction of larger peptides, however, is possible in systems where mosaic phages are produced by in vivo mixing the recombinant, peptide-containing, pVIII proteins with wild type pVIII (Felici et al, 1991; Greenwood et al, 1991; Willis et al, 1993).\",\n \"This enables the incorporation of the chimeric pVIII proteins at low density (tens to hundreds of copies per particle) on the phage surface interspersed with wild type coat proteins during the assembly of phage particles.\",\n \"Two systems have been used that enable the generation of mosaic phages; the “type 8+8” and “type 88” systems as designated by Smith (Smith, 1993).\",\n \"The “type 8+8” system is based on having the two pVIII genes situated separately in two different genetic units (Felici et al, 1991; Greenwood et al, 1991; Willis et al, 1993).\",\n \"The recombinant pVIII gene is located on a phagemid, a plasmid that contains, in addition to its own origin of replication, the phage origins of replication and packaging signal.\",\n \"The wild type pVIII protein is supplied by superinfecting phagemid-harboring bacteria with a helper phage.\",\n \"In addition, the helper phage provides the phage replication and assembly machinery that package both the phagemid and the helper genomes into virions.\",\n \"Therefore, two types of particles are secreted by such bacteria, helper and phagemid, both of which incorporate a mixture of recombinant and wild type pVIII proteins.\",\n \"The “type 88” system benefits by containing the two pVIII genes in one and the same infectious phage genome.\",\n \"Thus, this obviates the need for a helper phage and superinfection.\",\n \"Furthermore, only one type of mosaic phage is produced.\",\n \"The question arises, however, where one should introduce the second pVIII gene within the filamentous phage genome for efficient expression and genetic stability.\",\n \"The phage genome encodes 10 proteins (pI through pX) all of which are essential for production of infectious progeny (Felici et al, 1991).\",\n \"The genes for the proteins are organized in two tightly packed transcriptional units separated by two non-coding regions (Van Wezenbeek et al, 1980).\",\n \"One non-coding region, called the “intergenic region” (defined as situated between the pIV and pII genes) contains the (+) and the (−) origins of DNA replication and the packaging signal of the phage, enabling the initiation of capsid formation.\",\n \"Parts of this intergenic region are dispensable (Kim et al, 1981; Dotto et al, 1984).\",\n \"Moreover, this region has been found to be able to tolerate the insertion of foreign DNAs at several sites (Messing, 1983; Moses et al, 1980; Zacher et al, 1980).\",\n \"The second non-coding region of the phage is located between the pVIII and pIII genes, and has also been used to incorporate foreign recombinant genes as was illustrated by Pluckthun (Krebber et al, 1995).\",\n \"Regardless as to where a second pVIII gene is to be introduced, a major point for concern is the genetic stability of the ultimate vector and its derivatives.\",\n \"SUMMARY OF THE INVENTION The present invention is based on a critical examination of the attributes of the two non-coding regions of the fd filamentous phage as potential sites for insertion of a second recombinant pVIII gene and its genetic stability, resulting in the design and construction of an efficient “type 88” phage display expression system.\",\n \"The phage display expression system of the present invention includes a vector which is the DNA sequence of a filamentous phage into which a second pVIII gene, as well as DNA encoding a peptide of interest, are placed between the wild type pVIII and pIII genes.\",\n \"This allows production of type 88 phages displaying a peptide of interest with genetic stability and high copy number.\",\n \"The DNA encoding the second pVIII gene preferably uses alternative codons for encoding the amino acid residues of the pVIII protein.\",\n \"The native pVIII DNA sequence is separated from the recombinant pVIII sequence by a region encoding the wild type pVIII C-terminal domain and a terminator.\",\n \"The region encoding the wild type pVIII C-terminal domain is preferably designed to use alternative codons which are other than the native codons for encoding the same amino acids.\",\n \"In this way homologous recombination, slippage mechanisms and other genetic instabilities may be avoided.\",\n \"Preferably, the terminator at the end of the native pVIII DNA sequence is an HP terminator which is not native to the filamentous phage DNA.\",\n \"In a preferred embodiment of the present invention, a positive selection marker, such as the tetracycline or kanamycin resistant genes, is inserted between the native and recombinant pVIII genes.\",\n \"Preferably, a unidirectional promoter is substituted for the native bidirectional tetracycline resistant gene promoter.\",\n \"In preferred embodiments of the present invention, the native intergenic space between pIV and pII is maintained.\",\n \"The present invention further relates to the filamentous phages encoded by the DNA discussed above.\",\n \"Such phages may be of any type, such as fd, M13 and f1, although fd is preferred.\",\n \"The peptide designed to be displayed by the phage of the present invention can be any peptide which is desired to be presented, such as a specific antigen.\",\n \"Indeed, any protein or peptide can be displayed on the phage of the present invention.\",\n \"The peptide displayed on the phage may be an epitope of an antigen, which phage can be used therapeutically as a vaccine.\",\n \"Furthermore, the peptide displayed on the phage may be a single-chain antibody, which phage can be used for passive immunization.\",\n \"Both of these embodiments are described in WO 01/18169.Alternatively, one can create a library of phages into which are incorporated, at the same site, each of a set of oligonucleotides that encode all possible random peptides of a given length, or a large subset of that set.\",\n \"Such a large subset should preferably represent a fraction of the full set with the theoretical complexity of the full set of random polypeptides of a given length.\",\n \"Libraries of phages can also be prepared which incorporate all, or a large subset of all, of the overlapping oligonucleotides that represent all of the overlapping peptides of a given antigen and, thus, create a phage display pepscan.\",\n \"Scrambled pepscans (PCT application no.\",\n \"WO 98/20169) and phage display two hybrid systems (PCT application no.\",\n \"WO 98/20159) may also be prepared by this technique.\",\n \"The phage display libraries of the present invention can be used in screening for molecules which bind to a particular displayed peptide of interest or in screening the peptides displayed on a library of phages to see which bind to a specific molecule of interest, all as is well known with respect to prior art phage display libraries.\",\n \"Once a molecule or peptide of interest is identified by means of the screen using the phages or phage display libraries of the present invention, the peptide or molecule so identified may be produced in a conventional manner.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS FIGS.\",\n \"1A and 1B are a comparison between fd (FIG.\",\n \"1A) and fd-tet (FIG.\",\n \"1B).\",\n \"Opened arrows indicate the phage-transcriptional units.\",\n \"The darkened segment in FIG.\",\n \"1B indicates the tet fragment introduced into the BamH I site of fd at position 6272 (bold in FIG.\",\n \"1A) in the intergenic region (IG).\",\n \"FIGS.\",\n \"2A-2B show a deletion of 2.8 kb in fd-tet.\",\n \"FIG.\",\n \"2A is an uncut double-stranded DNA of fd-tet isolated from K802 (lane 1) or K91KAN (lane 2) and run on 0.8% agarose gel.\",\n \"Uncut double-stranded DNA of wild type fd was used for comparison (lane 3).\",\n \"M: DNA ladder mix (numbers equal base pairs).\",\n \"FIG.\",\n \"2B shows Msc I digests of fd-tet derived from K802 (lane 1) or K91KAN (lane 2) which were run on 0.8% agarose gel.\",\n \"Msc I digest of fd was used for comparison (lane 3).\",\n \"M: λ Hind III digest (numbers equal base pairs).\",\n \"Note that the lower band in lane 2 in FIG.\",\n \"2B, corresponding to the deleted product, is at least as intense as the upper band corresponding to the linear full-length fd-tet .\",\n \"FIG.\",\n \"3 shows the genetic instability of the inserted tet fragment.\",\n \"SnaB I/Ava I digests of fd-tet derived from K802 (lane 1) or K91KAN (lane 2).\",\n \"SnaB I/Ava I digest of fd was used for comparison (lane 3).\",\n \"BspH I/Hind III digests of fd-tet derived from K802 (lane 4) or K91KAN (lane 5:).\",\n \"BspH I/Hind III digest of fd DNA was used for comparison (lane 6).\",\n \"The arrow indicates the deleted products (lane 2 and 5) corresponding in size to the linear full-length fd (lane 3 and 6).\",\n \"M: DNA ladder mix.\",\n \"Agarose gel concentration was 0.8%.\",\n \"FIGS.\",\n \"4A-4B show that the deletion is not a precise reversion to fd.\",\n \"FIG.\",\n \"4A is BamH I digests of fd-tet DNA derived from K802 (lane 1) or K91KAN (and 2).\",\n \"BamH I digest of fd was used for comparison (lane 3).\",\n \"Note that the pattern obtained in lane 2 is identical to the Msc I digest in FIG.\",\n \"2B, lane 2.M: DNA ladder mix.\",\n \"Agarose gel concentration was 0.8%.\",\n \"FIG.\",\n \"4B is BstY I digests of fd-tet derived from K802 (lane 1) or K91KAN (lane 2) run on 0.8% agarose gel.\",\n \"BstY I digest of fd was used for comparison (lane 3).\",\n \"M: DNA ladder mix.\",\n \"Note the relative drop in intensity for the lowest band in FIG.\",\n \"4B, lane 2.FIG.\",\n \"5 shows the genome of the fd-tet phage.\",\n \"Genes (darkened segments), promoters (bent arrows), terminators (stem and loop), and selected restriction sites relevant to this study are shown.\",\n \"Strong promoters and terminators are in black.\",\n \"Weak promoters and terminators are in gray.\",\n \"The wild type fd genes (pI through pX) are packed in two transcriptional units.\",\n \"The inserted tet fragment contains two genes; tetA encoding for the tetracycline resistance protein and tetR encoding for the repressor protein regulating the expression of the tetA gene.\",\n \"The overlapping promoter/terminator, between pVIII and pIII genes, are inseparable.\",\n \"Moreover, the −35 box of this promoter is situated in the C-terminal coding region of pVIII gene (see text).\",\n \"Note the bi-directional promoter driving tetR and teta transcription.\",\n \"FIGS.\",\n \"6A-6C show the construction of two tandem pVIII genes.\",\n \"FIG.\",\n \"6A is a detailed comparison between the wild type pVIII gene (the contiguous central portion being SEQ ID NO:1) and the recombinant pVIII gene (the contiguous central portion being SEQ ID NO:3), designated pVIIISTS.\",\n \"The amino acid sequence shown to be encoded by SEQ ID NO:1 is SEQ ID NO:2.The amino acid sequences shown to be encoded by SEQ ID NO:3 are SEQ ID NOs:4 and 5.In the pVIII gene, the GAC codon for Asp at position 4 of the wild type pVIII is deleted and replaced by a sequence of 62 bp containing two stop codons and trpA transcription terminator flanked by two Sfi I sites (“STS” insert) (nucleotides 13-74 of SEQ ID NO:3).\",\n \"FIG.\",\n \"6B is the pGEM-t(p8STS) construct.\",\n \"The fragment containing the pVIIISTS gene, preceded by a SnaB I restriction site and followed by downstream pIII gene, was introduced into the pGEM-T vector.\",\n \"Black segments are sequences corresponding to the pGEM-T vector.\",\n \"The 62 bp “STS” insert is indicated.\",\n \"FIG.\",\n \"6C is the pGEM-t(pB8STS) construct.\",\n \"This construct contains the wild type pVIII and the modified pVIIISTS genes arranged as tandem repeats.\",\n \"The fragment containing the wild type pVIII gene starting from the SnaB I site and ending just beyond the overlapping promoter/terminator was introduced into the SnaB I site of pGEM-T(p8STS) thus destroying the former site while concomitantly introducing a new SnaB I site upstream to the wild type pVIII gene.\",\n \"Furthermore, additional unique sites were introduced between the two pVIII genes.\",\n \"FIG.\",\n \"7 is the genome of ftac88 vector.\",\n \"Most of the tetR gene was exchanged for a linker containing the Ava II site, thus eliminating the second SnaB I site previously situated in this region (for comparison see FIG.\",\n \"5).\",\n \"The modified pVIIISTS gene is situated between the duplicated overlapping promoter/terminator structures, without disrupting the two-phage transcriptional units.\",\n \"Note the pVIIISTS gene is under the control of a tac promoter (Ptac).\",\n \"FIG.\",\n \"8 shows the binding of mAb GV4H3 to phages expressing the GV4H3 epitope.\",\n \"Equal amounts of phages were applied in duplicates to a nitrocellulose membrane filter and reacted with the GV4H3 mAb.\",\n \"The GV4H3 epitope displaying phages, ftac88(4H3) and fth1(4H3), showed strong signals.\",\n \"Phage ftac88 and fth1 were used as negative controls.\",\n \"FIG.\",\n \"9 shows the deletion of the recombinant pVIII gene in ftac88(4H3).\",\n \"SnaB I/BamH I digests of ftac88 derived from DH5α (lane 1) or DH5αF′ (lane 2) were run on 0.8% agarose gel.\",\n \"SnaB I/BamH I digests of ftac88 (4H3) isolated from DH5α (lane 3) or DH5αF′ (lane 4) are also shown.\",\n \"M: DNA ladder mix.\",\n \"The upper arrow indicates the tet insert deleted product and the lower arrow indicates the recombinant pVIII gene deleted fragment.\",\n \"FIG.\",\n \"10 shows the fth1 vector (SEQ ID NO:29).\",\n \"The general scheme of the fth1 vector is shown in scale illustrating the phage genes (opened segments), the reconstituted intergenic region (IG) and the genes introduced between the wild type pVIII and pIII genes (gray segments).\",\n \"Below, detail of the pIX to the middle of pIII is provided (not in scale).\",\n \"The darkened segments represent the modified wild type pVIII 3′-end and the −35 box of the pIII promoter (see also FIG.\",\n \"11).\",\n \"For details see text.\",\n \"FIGS.\",\n \"11A-11C show the intermediate vectors constructed for the formation of fth1 vector.\",\n \"FIG.\",\n \"11A is a detail of the segment of fd-tet containing the pIX, pVIII, and pIII genes.\",\n \"The segment in pVIII indicating the −35 box of the pIII promoter is shown in black situated in the 3′ region of the wild type pVIII open reading frame.\",\n \"FIG.\",\n \"11B is the intermediate vector-1 (IV-1).\",\n \"The −35 box of the pIII promoter was separated from the wild type pVIII gene by inserting a small segment reconstituting the 3′ region and introducing multiple cloning sites.\",\n \"FIG.\",\n \"11C is IV-4.A novel HP terminator (tHP) is situated downstream to the wild type pVIII gene terminating its transcription followed by the tetracycline resistance gene (tetA) under the control of the kan promoter (Pkan).\",\n \"FIG.\",\n \"12 shows the genetic stability of fth1.BstN I digests of fth1 derived from DH5α (lane 2) or DH5αF′ (lane 3).\",\n \"BstN I digests of fth1(4H3) derived from DH5α (lane 4) or DH5αF′ (lane 5).\",\n \"BstN I digest of fd was used for comparison (lane 1).\",\n \"M: DNA ladder mix.\",\n \"Agarose gel concentration was 0.8%.\",\n \"DETAILED DESCRIPTION OF THE INVENTION Type 88 display systems are in essence a phage genome that contains two protein VIII genes.\",\n \"One of the genes is the wild type pVIII, whereas the second is modified so as to enable expression of a peptide (random or pre-defined peptide, or even a single-chain antibody (scFv)).\",\n \"The problem with known type 88 phage display systems is genetic instability.\",\n \"After a number of generations, the phages tend to stop production of the inserted polypeptide of interest and no longer display such polypeptide.\",\n \"The novel constructs of the present invention solve this problem.\",\n \"Furthermore, in prior art systems, in which the selectable marker has been introduced into the intergenic region, some of the functions of the replication of the phage are obstructed, and the actual amount of phage DNA being produced in a liter of culture may be no more than 20-50 μg of DNA.\",\n \"However, with the construct of the present invention, high copy numbers may be obtained.\",\n \"Thus, as much as 500-1000 μg of DNA can be readily purified from a liter of culture.\",\n \"In the vector of the present invention, the sequence of a filamentous phage is manipulated so as to insert a second pVIII gene and the gene encoding the polypeptide of interest between the wild type pVIII and pIII genes.\",\n \"In a preferred embodiment, the intergenic region between pII and pIV is not modified and retains the native sequence, thus allowing for the high copy number to be obtained.\",\n \"Any positive selection marker may also be introduced between the wild type pVIII and pIII genes.\",\n \"In the wild type filamentous phage DNA, the −35 box necessary for pIII expression is positioned to overlap with the C-terminal coding region of the pVIII protein.\",\n \"Thus, the −35 regulatory sequence is upstream of the C-terminal coding region and terminator of the pVIII protein.\",\n \"So as not to disrupt the expression of the pIII protein, the vector of the present invention preferably maintains the genuine C-terminal/−35 box at the 5′ end of the pIII gene.\",\n \"A new C-terminal region for the native pVIII gene is introduced upstream thereof.\",\n \"The second pVIII region and the gene encoding the polypeptide of interest is introduced between the newly-introduced terminator for the native pVIII gene and the C-terminal/−35 box 5′ to the pIII gene.\",\n \"In order to prevent possible homologous recombination, slippage mechanisms, or other genetic instabilities, it is preferable that the DNA encoding the second pVIII gene use alternative codons which are other than the native codons for encoding amino acid residues of the pVIII protein.\",\n \"Enough alternative codons should be inserted therein so as to avoid stretches longer than 30 bases of the native sequence, preferably no longer than 20 such bases.\",\n \"Similarly, the region encoding the wild type pVIII C-terminal domain should also use alternative codons in sufficient quantity to avoid stretches longer than 30 native bases, and preferably to avoid stretches longer than 20 such bases.\",\n \"The positive selection marker is preferably inserted between the native and recombinant pVIII genes.\",\n \"The present invention further comprehends a vector in which the second pVIII gene is introduced just downstream to the −35 promotor of the pIII gene (and thus driven by the pIII promotor and not the tac promotor of fth1 or ftac88 as described).\",\n \"In such a situation, due to the presence of the transcription terminator in the STS stuffer of the recombinant pVIII, phages that do not incorporate an insert for the polypeptide of interest will not be produced at all.\",\n \"This is due to the fact that the terminator prevents the expression of the pIII, pVI and pI proteins.\",\n \"The pI protein is essential for phage assembly.\",\n \"In the situation where an insert is cloned in place of the stuffer, the recombinant phage is made.\",\n \"If such an insert is not incorporated, then transcription is terminated and no phage is produced.\",\n \"While the fd filamentous phage is used in the present examples and is the preferred phage sequence for use in the present invention, it should be understood that all filamentous phages are very similar and have the same gene organization (Model et al, 1988).\",\n \"Thus, the principles of the present invention can be applied to any of the filamentous phages, such as M13, f1 and others.\",\n \"The native pVIII DNA sequence is preferably separated from the second pVIII DNA sequence by a terminator which is not native to the filamentous phage DNA, preferably the HP terminator.\",\n \"In the preferred embodiment of the present invention, the native, unique, overlapping pVIII terminator/pIII promoter is maintained at its native position at the N-terminal portion of the pIII gene.\",\n \"This includes the pVIII C-terminal region/pIII −35 box.\",\n \"However, to further avoid tandem repeats, the pVIII C-terminal region, which is present at this point, is preferably designed with alternative codon usage to avoid stretches of longer than 30 native bases, and preferably to avoid stretches longer than 20 native bases.\",\n \"While the DNA molecule of the present invention is described as having a recombinant DNA sequence encoding the pVIII protein and DNA encoding a polypeptide of interest other than pVIII which is inserted between the wild type pVIII and pIII genes, this language encompasses the presence of DNA encoding a peptide of interest within the sequence encoding the pVIII protein.\",\n \"Preferably, the DNA sequence encoding the peptide of interest is present at the N-terminal region of the pVIII gene.\",\n \"The peptide of interest and the pVIII gene will be expressed as a fusion protein in which the peptide of interest is expressed on the surface of the filamentous phage.\",\n \"While the selectable marker disclosed herein is the tetracycline resistance gene, those of ordinary skill in the art will understand that any selectable marker can be used for this purpose, such as the kanamycin resistance gene.\",\n \"Furthermore, as the native promoter for the tetracycline resistance gene is bidirectional, in a preferred embodiment of the present invention, this promoter is replaced by a unidirectional promoter, such as the kanamycin promoter.\",\n \"The present invention also comprehends a vector which may be used as an intermediate for producing a vector containing the DNA encoding the peptide of interest.\",\n \"In this intermediate vector, the DNA of the phage has inserted therein a recombinant DNA sequence that includes a multiple cloning site.\",\n \"The multiple cloning site has a series of restriction sites that do not otherwise appear in the DNA of the phage into which the recombinant DNA has been inserted.\",\n \"The recombinant DNA sequence is designed and inserted such that the multiple cloning site is placed between a terminator for the wild type pVIII gene and an initiator for the wild type pII gene.\",\n \"The multiple cloning site may then be used to readily insert DNA sequences encoding the foreign peptide of interest to be displayed on the phage.\",\n \"A positive selection marker, as has been discussed elsewhere in the present specification, may be present in the multiple cloning site in order to simplify insertion only of the DNA encoding the peptide of interest so as to arrive at the final product.\",\n \"The second pVIII gene may also be present in the multiple cloning site.\",\n \"Instead of a second pVIII gene, any recombinant bacteriophage gene can be present in the multiple cloning site.\",\n \"For example, it would be useful to produce a system in which two pIII genes are present.\",\n \"This would allow the production of both wild type and recombinant pIII genes in the same phage.\",\n \"It is expected that, by modifying the efficiency of expression, one can create a situation in which only one recombinant pIII protein is expressed per phage.\",\n \"The normal situation is that five pIII proteins are present per phage.\",\n \"When all are recombinant, then the binding is affected by the multivalency of the five copies which leads to enhanced avidity of binding and can be misleading if one wants to be precise in evaluating real affinity (as opposed to avidity which benefits from multiple binding reactions).\",\n \"Similarly, any of the native proteins (pI through pX) can be duplicated in the multiple cloning site for whatever reason.\",\n \"The recombinant DNA sequence, which is inserted into the DNA of the filamentous phage, is designed and inserted such that the multiple cloning site is placed between a terminator for the wild type pVIII gene and an initiator for the wild type pIII gene.\",\n \"As discussed hereinabove, preferably the wild type C-terminal/−35 box is retained at the 5′ end of the pIII gene, and a new C-terminal region for the native pVIII gene is introduced upstream of the multiple cloning site.\",\n \"Preferably, in the intermediate vector, in the region between flanking restriction enzyme sites, a trpA transcription terminator is disposed, although any random combination of bases can appear between these flanking restriction enzyme sites.\",\n \"When this intermediate is to be used for production of the vector to be used to make a phage displaying the peptide of interest, DNA encoding the peptide of interest is merely substituted for the DNA of the intermediate vector between the two flanking unique restriction enzyme sites.\",\n \"Since the recombinant DNA sequence inserted into the phage in order to create the multiple cloning site must include a region homologous to the N-terminal portion of the pIII gene, it is possible in this same insert to include a DNA sequence encoding another foreign peptide incorporated into the pIII gene at the N-terminal region thereof.\",\n \"This may be used, for example, to produce a phage that displays a library of random peptides on either pIII or pVIII and a marker on the other, such as green fluorescence protein.\",\n \"This would make the selection of the bound phage easier or amenable to high throughput screening.\",\n \"Also encompassed by the present invention is the recombinant DNA construct which is produced and used for insertion of the multiple cloning sites, into the phage DNA so as to appear between the pVIII and pIII genes.\",\n \"This construct is designed so as to replace the wild type sequence between restriction sites which are already present in the phage, such as the SnaBI in the pIX gene and the BamHI in the pIII gene.\",\n \"The ends of the construct are designed so as to be compatible with the restriction enzyme cleavage sites left after removal of the wild type sequence such that efficient ligation of the construct into the vector may be obtained, all as is well known to those of ordinary skill in the art.\",\n \"The novel DNA vectors of the present invention and the novel type 88 filamentous phages encoded thereby may be used in the same manner that known peptide display phages have been known to be used in the prior art, except that they are genetically stable, have high copy number, and produce high titres of phages.\",\n \"The vector can be used to express any protein or peptide.\",\n \"In the situation in which a discrete sequence of peptide is known which is desired to be expressed, then such a known peptide can readily be incorporated into the vector so as to produce high titres of phage displaying the desired discrete amino acid sequence.\",\n \"An example of the use of a discrete sequence of peptide displayed on a phage is represented by the disclosure of PCT application no.\",\n \"WO 01/18169, the entire contents of which are hereby incorporated herein by reference.\",\n \"This publication discloses the utility as a vaccine of phages displaying a particular antigenic epitope so as to raise antibodies against that epitope.\",\n \"Also disclosed therein are phages displaying single-chain antibodies which can be directly used for passive immunization.\",\n \"Thus, phages displaying such antigens or single-chain antibodies for use in the particular applications of WO 01/18169, or any related application, are considered to be comprehended by the present invention when the antigen or single-chain antibody is displayed on a phage in accordance with the present invention.\",\n \"Additionally, as is known in the art, libraries can be prepared that incorporate into the same site of the vector a set of oligonucleotides that code all possible random peptides of a given length, or a large subset of that set, as is known in the art.\",\n \"Such a large subset should preferably represent a fraction of the full set with the theoretical complexity of the full set of random polypeptides of a given length.\",\n \"Such phage display libraries can be used to screen for amino acid combinations which will bind to a given target molecule.\",\n \"Once a phage which binds to the desired target molecule is found, the peptide insert in that phage can be determined and molecules containing that peptide can be produced.\",\n \"Such molecule would be expected to bind the target molecule.\",\n \"The binding of the target molecule to its native receptor or ligand may be disrupted.\",\n \"This is useful when it is desired to prevent the signaling which occurs when the target molecule is bound to its native ligand or receptor, all as is well known in the art of phage display libraries.\",\n \"Furthermore, libraries can be prepared which incorporate the set of all of the overlapping oligonucleotides that represent all the overlapping peptides of a given antigen, or a large subset thereof, and, thus, create a phage display pepscan.\",\n \"A scrambled pepscan, such as the combinatorial scrambled vaccines described in PCT application no.\",\n \"WO 98/20169, the entire contents of which are hereby incorporated herein by reference, and the phage display two hybrid systems, such as are described in PCT application no.\",\n \"WO 98/20159, the entire contents of which are hereby incorporated by reference, may also be prepared using the novel phages of the present invention.\",\n \"Any of such phage display libraries can be used in screen assays as is known in the art.\",\n \"In any such screening assay, the peptide that is found can then be produced and used for its intended purpose, again, as is well known in the prior art.\",\n \"An example of such a screen would be to use a library in accordance with the present invention against a receptor.\",\n \"In this way one might be able to select for a peptide that mimics the native ligand.\",\n \"This peptide could then be produced using standard Merrifield synthesis, and the synthetic peptide may be used as either a lead peptide for further development or directly as a modulator of the receptor being studied.\",\n \"For example, a random phage display peptide library may be used to screen against HIV gp120.In this way, one might be able to select for a peptide that binds to gp120 at precisely the CD4 binding site.\",\n \"Such a peptide would be expected to bind to the virus and to prevent the association of the virus to the CD4 receptor and, thus, prevent infection.\",\n \"One could also produce a library derived from CD4 itself and hope to discover a fragment of CD4 that binds to gp120 and acts as a decoy.\",\n \"All possibilities stem from the fact that the library is produced in the genetically stable expression system of the present invention.\",\n \"As to the selectable marker, a particular advantage of placing this marker between the two pVIII genes is to create a situation that if, for whatever reason, there is a genetic recombination based on the limited homology of the native pVIII gene and the recombinant pVIII gene, then the selectable marker would be excised and lost as well.\",\n \"Running an experiment in the presence of the antibiotic would result in loss of those recombinants that have lost their resistance gene.\",\n \"Other markers that could be used in place of the tetracycline resistance gene would be the ampicillin resistance gene, β-lactamase or chloramphenicol resistance gene.\",\n \"As stated hereinabove, it is also possible within the scope of the present invention to also insert an exogenous gene between the pIII gene and its promoter.\",\n \"By doing so, the phage can display a library of random peptides on either pIII or pVIII and a marker on the other, such as green fluorescence protein.\",\n \"This would make the selection of the bound phage easier or amenable to high throughput screening.\",\n \"The new exogenous gene can either be inserted directly between the pIII gene and its promoter or a second pIII gene can be inserted similarly to that discussed in the present invention for the second pVIII gene in order to ensure that the native pIII gene will always be produced along with the second pIII gene and the peptide of interest.\",\n \"It is not necessary to insert a separate pIII gene as, in contrast to pVIII, pIII can tolerate large inserts.\",\n \"The following examples are directed to preferred embodiments in accordance with the present invention and show how to make and use the present invention.\",\n \"Those of ordinary skill in the art will understand, however, that these examples do not detract from the breadth of the present invention as described herein and that appropriate substitutions can be made without engaging in undue experimentation by those of ordinary skill in the art.\",\n \"Materials and Methods Bacterial Strains, Phages, Reagents, and General Techniques E. coli strains used in this study were the following.\",\n \"K802: F− e14−(McrA−) lacY1 or Δ(lac)6 supE44 galK2 galT22 rfbD1 metB1 mcrB1 hsdS3(rK− mK−).\",\n \"K91KAN: a derivative of K91 (Hfr-Cavalli, thi) in which the “mini-Kan hopper” element was inserted in the lacZ gene of K91 rendering this strain kanamycin resistant (Parmley et al, 1988).\",\n \"DH5α: F− endA1 hsdR17(rK− mK+) supE44 thi-1 recA1 gyrA(Nalr) relA1 Δ(lacIZYA-argF)U169 deoR (Φ80dlacΔ(lacZ)M15).\",\n \"DH5αF′: F′ endA1 hsdR17(rK− mK+) supE44 thi-1 recA1 gyrA(Nalr) relA1 Δ(lacIZYA-argF)U169 deoR (Φ80dlacΔ(1acZ)M15).\",\n \"JM109: F′[traD36 lacIq Δ(lacZ)M15 proA+B+] e14−(McrA−) Δ(lac-proAB) thi gyrA96(Nalr) endA1 hsdR17(rK− mK+) relA1 supE44 recA1.XL1-Blue: F′[::Tn10 proA+B+ lacIq Δ(lacZ)M15] recA1 endA1 gyrA96(Nalr) thi hsdR17(rK− mK+) supE44 relA1 lac.\",\n \"MC1061: F− araD139 Δ(ara-leu)7696 galE15 galK16 Δ(lac)X74 rpsL(Strr) hsdR2(rK− mK+) mcrA mcrB1.NM554: F− araD139 Δ(ara-leu)7696 galE15 galK16 Δ(lac)X74 rpsL(Strr) hsdR2(rK− mK+) mcrA mcrB1 recA13.The wild type fd filamentous phage and fd-tet vector were kindly provided by G. P. Smith (University of Missouri, Columbia, Mo.).\",\n \"The M13K07 phage was purchased from New England BioLabs Inc., MA (NEB).\",\n \"DNA was isolated using the alkaline lysis procedure and purified on a cesium chloride gradient as described previously (Stern et al, 1998).\",\n \"All the restriction enzymes used were purchased from NEB and the digestions were performed following the manufacturer's instructions.\",\n \"The monoclonal antibody GV4H3 was produced from a Balb/c mouse immunized with the HIV-1 envelope protein gp120 (Denisova et al, 1996).\",\n \"Oligonucleotides The oligonucleotides used in this study were the following.\",\n \"ON1: 5′-GCCTTCGTAGTGGCATTACG-3′ (SEQ ID NO: 6) ON2: 5′-AGCCCGCTCATTAGGCGGGCTTCATTACCGGCCACGTCGGCCAC (SEQ ID NO: 7) CCTCAGCAGCGAAAGAC-3′ ON3: 5′-CGGTAATGAAGCCCGCCTAATGAGCGGGCTTTTTTTTGGCCTCT (SEQ ID NO: 8) GGGGCCGATCCCGCAAAAGCGGCC-3′ ON4: 5′-GGTCAGACGATTGGCCTTG-3′ (SEQ ID NO: 9) ON5: 5′-GAGCTCGCTAGCGCTCGAGCCAAAAAAAAAGGCTCCAAAAGG-3′ (SEQ ID NO: 10) ON6: 5′-CTAGAGCAGGGTCCAGCTAA-3′ (SEQ ID NO: 11) ON7: 5′-CTGGACCCTGCT-3′ (SEQ ID NO: 12) ON8: 5′-TCGAGCTGTTGACAATTAATCATCGGCTCGTATAATGTGTGGAA (SEQ ID NO: 13) TTGTGAGCGGATAACAATTGAGCT-3′ ON9: 5′-CAATTGTTATCCGCTCACAATTCCACACATTATACGAGCCGATG (SEQ ID NO: 14) ATTAATTGTCAACAGC-3′ ON10: 5′-CCGTGCATCTGTCCTCGTTC-3′ (SEQ ID NO: 15) ON11: 5′-CAGGCTTAAGCATCGACGTCTTATCAAGACGCCTTGCTTGTAA (SEQ ID NO: 16) ACTTTTTGAATAACTTGATACCGATAGTTGCGC-3′ ON12: 5′-GTCTTGATAAGACGTCGATGCTTAAGCCTGGCTAGCCATCAGAT (SEQ ID NO: 17) CTGAGTCGGCCGCTGTTTAAGAAATTCACCTCG-3′ ON13: 5′-GCCGTACCGCTAGCATTAGAAAAACTCATCGAGC-3′ (SEQ ID NO: 18) ON14: 5′-GCTTCCTGACAGGAGGCCGTTTTGTTTTGCAGCCCACCTGAGCT (SEQ ID NO: 19) CCCAGCTTAAGGTGTCTCAAAATCTCTGATG-3′ ON15: 5′-GGCTGAGGGACGTCGAGGGCATGCGTACCCGATAAAAGCGGCTT (SEQ ID NO: 20) CCTGACAGGAGGCCG-3′ ON16: 5′-AGCCTTTCAGGTCAGAAGGG-3′ (SEQ ID NO: 21) ON17: 5′-ATTCATGCCGGAGAGGGTAG-3′ (SEQ ID NO: 22) ON18: 5′-CATAACACCCCTTGTATTACTG-3′ (SEQ ID NO: 23) ON19: 5′-GCTTACATAAACAGTAATACAAGGGGTGTTATGAATAGTTCGAC (SEQ ID NO: 24) AAAGATCGC-3′ ON20: 5′-GGATCGAGAGCTAGCATAACTAAGCACTTGTCTCCTG-3′ (SEQ ID NO: 25) ON21: 5′-TGCCGGCGCTCCCGCTGGTTTCGCTATCCTGTCAGCACCCGGGT (SEQ ID NO: 26) CTG-3′ ON22: 5′-ACCCGGGTGCTGACAGGATAGCGAAACCAGCGGGAGCGCCGGCA (SEQ ID NO: 27) CGT-3′ Vector Construction In this study two novel vector-systems were constructed.\",\n \"The rationale for their compositions and structures is described in the Results.\",\n \"Detailed diagrams of the vectors and sub-structures are given in the Figures as indicated.\",\n \"The following are the details of the specific steps taken to construct the vectors.\",\n \"Construction of ftac88 The ftac88 vector (for detailed map see FIG.\",\n \"7) was constructed through the following steps.\",\n \"First, the recombinant pVIII gene, designated pVIIISTS (for orientation see FIG.\",\n \"6A), was generated by introducing a 62 bp insert three codons downstream to the leader peptide using the “SOEing” PCR mutagenesis (Horton et al, 1990).\",\n \"For this, four oligonucleotides were used.\",\n \"ON1 and ON2 were used to PCR amplify from fd a 169 bp fragment.\",\n \"ON3 and ON4 were used to PCR generate from fd a 899 bp fragment.\",\n \"ON2 and ON3 each contain 5′-extension corresponding to the 62 bp insert.\",\n \"Thus, the resulting PCR fragments contain an identical 30 bp stretch of the novel sequence.\",\n \"The two fragments were purified from agarose gel, mixed and used as a combined template for PCR amplification using the oligonucleotides ON1 and ON4 to generate the final 1071 bp product containing the modified pVIII gene, part of pIII gene, and the flanking BamH I-SnaB I sites.\",\n \"The PCR reaction was performed by using the Taq polymerase which adds adenine to the 3′ ends of PCR products and thus the resulting 1071 bp fragment was directly ligated with the linearized pGEM-T vector (Promega Corporation, WI) containing complementary 3′ thymidine overhangs, generating the pGEM-T(p8STS) construct (see FIG.\",\n \"6B).\",\n \"Next, the wild type pVIII gene was PCR amplified from fd using the oligonucleotides ON1 and ON5.ON5 has a 5′ extension that contains the restriction sites Xho I, Nhe I and Sac I.\",\n \"The PCR reaction was performed using the Pwo Polymerase (Boehringer Mannheim), resulting in blunt-ended 335 bp fragment from the SnaB I site to just beyond the overlapping promoter/terminator followed by the ON5 introduced restriction sites.\",\n \"The resulting fragment was ligated with the blunt-ended SnaB I linearized pGEM-T(p8STS) construct generating the pGEM-T(p88STS) construct (see FIG.\",\n \"6C).\",\n \"To remove the SnaB I site in tetR gene, the fd-tet vector (for detailed map see FIG.\",\n \"5) was digested with Xba I and BstX I restriction enzymes and the 8458 bp fragment was purified from agarose gel.\",\n \"The purified fragment was ligated with a linker containing the Ava II site produced by annealing the two complementary oligonucleotides ON6 and ON7.To exchange the SnaB I-BamH 1952 bp segment of fd-tet for the SnaB I-BamH 11378 bp segment of pGEM-T(p88STS) construct containing both the wild type and recombinant pVIII genes, the Ava II containing fd-tet derivative was digested with SnaB I/BamH I and the 7519 bp fragment was purified from agarose gel.\",\n \"In the same manner, the pGEM-T(p88STS) construct was digested with SnaB I/BamH I and the released 1378 bp fragment was purified from agarose gel.\",\n \"These two fragments were ligated to generate an intermediate vector.\",\n \"This intermediate vector was digested with Sac I and Xho I restriction enzymes and the 13 bp stuffer was removed by applying the DNA digest on a chroma-spin™ column (Clontech Laboratories, Inc., Palo Alto, Calif.).\",\n \"The linearized intermediate vector was ligated with a linker containing the tac promoter produced by annealing the two complementary oligonucleotides ON8 and ON9, generating the ftac88 vector.\",\n \"Construction of fth1 The construction of the fth1 vector (for detailed map see FIG.\",\n \"10) was performed through a series of intermediate vectors.\",\n \"The intermediate vector-1 (IV-1, see FIG.\",\n \"11B) was constructed by introducing a DNA segment ten codons upstream to the stop codon of the wild type pVIII gene using the “SOEing” PCR mutagenesis described above.\",\n \"Two fragments of 519 bp and 827 bp were PCR generated from fd using the oligonucleotides ON10/ON11 and ON12/ON4 respectively.\",\n \"The two fragments were mixed and used as a combined template using ON10/ON4 to PCR generate the final 1316 bp fragment.\",\n \"This product was digested with SnaB I and BamH I and the resulting 1034 bp fragment was purified from agarose gel and used to replace the corresponding SnaB I-BamH I fragment of the Ava II containing fd-tet derivative described above for the construction of ftac88.Next, the IV-2 was constructed by introducing into the IV-1, downstream to the wild type pVIII gene, the novel HP transcription terminator followed by the kanamycin resistance gene.\",\n \"This was performed in order (i) to functionally separate the two phage-transcriptional units and (ii) to enable the reconstitution of the intergenic region while maintaining an alternative antibiotic selectable marker (i.e.\",\n \"kanamycin).\",\n \"The kanamycin resistance gene was PCR amplified from M13K07 using the oligonucleotides ON13 and ON14.Both oligonucleotides contain 5′ extensions enabling the incorporation of additional flanking sequences.\",\n \"The 3′ addition contained the Nhe I restriction site and the 5′ extension contained additional cloning sites adjacent to the kanamycin promoter and part of the HP terminator.\",\n \"The resulting 944 bp fragment was purified from agarose gel and used as a template for PCR amplification using the oligonucleotides ON13 and ON15.The latter has a 5′ extension containing the second part of the HP terminator and additional cloning sites ending with the Aat II site.\",\n \"Thus the resulting 984 bp fragment contains the entire HP terminator bracketed between multiple cloning sites followed by the kanamycin resistance gene.\",\n \"This product was digested with Nhe I and Aat II and the resulting fragment was purified from agarose gel.\",\n \"The purified fragment was ligated with Nhe I/Aat II digested IV-1 vector after removing the 20 bp stuffer by applying the vector digest on a chroma-spin™ column, ultimately generating IV-2.The IV-2 was used to reconstitute the intergenic region containing the origins of replication.\",\n \"This was performed by PCR amplifying the intergenic region from fd phage using the oligonucleotides ON16 and ON17.The resulting 1081 bp fragment containing the Msc I-Drd I segment was digested with Msc I and Drd I restriction enzymes.\",\n \"The 679 bp Msc I-Drd I product was purified from agarose gel and used to exchange the Msc I-Drd I fragment in IV-2 containing the tet fragment, generating IV-3.The open reading frame only of the kanamycin resistance gene was replaced by the open reading frame of the tetracycline resistance gene thus constructing the IV-4 vector (see FIG.\",\n \"11C).\",\n \"This was performed using the “SOEing” PCR.\",\n \"The oligonucleotides ON1 and ON18 were used to PCR amplify from IV-2 the 463 bp fragment containing the upstream sequence to the ATG initiating codon of the kanamycin gene.\",\n \"The oligonucleotides ON19 and ON20 were used to PCR amplify from fd-tet the 1265 bp fragment containing the open reading frame of the tetracycline resistance gene.\",\n \"The two fragments were mixed and used for PCR amplification of the 1698 bp fragment using the oligonucleotides ON1 and ON20.The PCR was designed to flank the resulting 1698 bp fragment between Nhe I and Sac I restriction sites enabling the digestion of this fragment with the corresponding enzymes.\",\n \"After purifying the digested fragment from agarose gel, it was used to replace the corresponding Nhe I-Sac I fragment of IV-3.The final step was to generate the synthetic recombinant pVIII gene using a panel of overlapping oligonucleotides (not shown).\",\n \"Part of the panel was used as templates and the other part as 5′-extended primers for PCR amplification thus generating separately small pieces of DNA.\",\n \"These small fragments were sequentially mixed and used as a combined template for the “SOEing” PCR ultimately generating the recombinant pVIIISTSh gene that subsequently was introduced between the Bgl II/Eag I sites of IV-4 producing the fth1 vector.\",\n \"Introduction of the GV4H3 Epitope into ftac88 and fth1 Vectors The vectors ftac88 and fth1 were digested with Sfi I restriction enzyme and the 49 bp stuffer containing the trpA terminator (for orientation see FIG.\",\n \"6a) was removed by applying the DNA digest on a chroma-spin™ column.\",\n \"The purified linear vector was ligated with GV4H3 epitope encoding linker produced by annealing the two complementary oligonucleotides ON21 and ON22.Dot Blot Analysis of GV4H3 Epitope Presenting Phages The phages were applied via a vacuum manifold to a nitrocellulose membrane filter.\",\n \"After blocking (5% evaporated spray dried skim milk 1.5% fat in Tris buffered saline (TBS)) for 1 hour, the membrane was washed briefly with TBS and incubated overnight with 1 μg/ml of GV4H3 mAb in TBS/5% milk at 4° C. with gentle rocking.\",\n \"After washing, the membrane was incubated with goat-anti mouse IgG/HRP conjugate diluted 1:5000 in TBS/5% milk for 1 hour at room temperature.\",\n \"The positive signals were detected by ECL (Amersham International plc, Buckinghamshire, England) immunodetection.\",\n \"Results The fd-tet vector of Smith (Zacher et al, 1980) was used as a starting point for our studies.\",\n \"The fd-tet Expression Vector Using the fd genome as a scaffold molecule, Zacher et al (1980) constructed the fd-tet phage, which contains the tetracycline resistance gene as a selectable marker (see FIG.\",\n \"1).\",\n \"The fragment coding for the tetracycline resistance was obtained from the transposon Tn10 (a 2775 bp fragment flanked between two Bgl II sites).\",\n \"This segment was introduced into the BamH I site of the intergenic region containing the origins of replication of the fd genome.\",\n \"The fd-tet vector has the following advantages: 1.fd-tet phages confer selectable tetracycline resistance to infected or transfected host bacteria.\",\n \"2.The tet fragment introduces unique restriction sites that can be exploited for cloning foreign DNA sequences.\",\n \"Thus, fd-tet was used, for example, as a cloning vector by exploiting the unique Hind III site, situated in the tet fragment, to clone Hind III digested phage X DNA (Zacher et al, 1980).\",\n \"Furthermore, Smith has reported the introduction of a second pVIII gene in the same region generating a “type 88” vector designated f88.4, and the successful production of pVIII mosaic phages (Zhong et al, 1994).\",\n \"However, a point for concern is the report that foreign DNA, cloned into the intergenic region of filamentous phages, tends to be unstable (Model et al, 1988; Sambrook et al, 1989; Vieira et al, 1987).\",\n \"Occasionally, parts of the cloned DNA are deleted and the rate of the deletion event tends to be directly correlated with the size of the inserted fragment.\",\n \"Therefore, the question of the genetic stability of DNA inserts cloned into the intergenic region was examined.\",\n \"Genetic Instability in fd-tet Vector Purified fd-tet dsDNA generated two distinct electrophoretic patterns on agarose gels when isolated from two different E. coli strains (K802 and K91KAN).\",\n \"The DNA isolated from K802 exhibited the supercoiled and relaxed forms of the fd-tet genome (FIG.\",\n \"2A, lane 1) whereas the DNA isolated from K91KAN contained the supercoiled and relaxed forms of two distinct DNAs (FIG.\",\n \"2A, lane 2).\",\n \"One DNA corresponded to the fd-tet genome while the second compared well with the parent fd genome (FIG.\",\n \"2A, lane 3).\",\n \"The two purified fd-tet preparations and fd were then digested with the Msc I restriction enzyme that should generate full length linear DNA for both fd and fd-tet (the Msc I restriction site is unique in both fd and fd-tet, see FIG.\",\n \"1).\",\n \"Digestion of fd-tet from K802 and fd each generated only one band with the expected linear sizes of 9183 bp and 6414 bp respectively (FIG.\",\n \"2B, lanes 1 and 3).\",\n \"However, digestion of fd-tet derived from K91KAN generated two bands, one indistinguishable in mobility from the linear fd-tet and the second corresponding in size to linear fd (FIG.\",\n \"2B, lane 2).\",\n \"This indicates that for a substantial amount of DNA, a deletion of approximately 2.8 kb occurred when the fd-tet genome was produced in K91KAN bacteria.\",\n \"Two points, therefore, must be addressed: 1.What DNA sequences are lost through the deletion process?\",\n \"2.How are the differences in the bacterial strains related to the apparent genetic instability?\",\n \"Deletion of the tet Fragment As the deleted DNA was 2.8 kbp, it was postulated that it might correspond to the tet-fragment (2775 bp) residing in the intergenic space of fd-tet (see FIG.\",\n \"1).\",\n \"Therefore, the SnaB I/Ava I digests of fd-tet DNA derived from both bacterial strains as before were compared.\",\n \"The SnaB I site is unique in fd, however, it appears twice in fd-tet while the Ava I site exists only in the tet fragment of fd-tet.\",\n \"The SnaB I/Ava I double digestion of fd-tet isolated from K802 generated three fragments with the expected sizes: 4381 bp, 2690 bp and 2112 bp (FIG.\",\n \"3, lane 1).\",\n \"The double digestion of the K91KAN derived fd-tet DNA, however, generated four DNA fragments (FIG.\",\n \"3, lane 2); the three fragments corresponding to fd-tet and an additional fragment corresponding to the deletion product similar in size to a single cut full length 6414 bp fd (FIG.\",\n \"3, lane 3, compare also with FIG.\",\n \"2B).\",\n \"This indicates that the SnaB I-Ava I fragment of the tet insert in the intergenic region was deleted from fd-tet.\",\n \"This conclusion was further supported by BspH I/Hind III digestion of fd-tet derived from both bacteria.\",\n \"The Hind III site is unique to fd-tet (see FIG.\",\n \"1).\",\n \"The BspH I site appears only once in fd and twice in fd-tet (see FIG.\",\n \"1, the second BspH I site is situated in the tet fragment).\",\n \"The BspH I/Hind III digestion of fd-tet DNA isolated from K802 generated the expected three fragments of 4676 bp, 2366 bp, and 2141 bp (FIG.\",\n \"3, lane 4).\",\n \"However, the BspH I/Hind III double digestion of fd-tet DNA isolated from K91KAN generated not only the three fd-tet derived fragments but also a fourth fragment (approximately 6.4 kb) corresponding in size to single cut full-length fd (FIG.\",\n \"3, lane 5, note arrow) derived from the deletion product.\",\n \"This not only confirms the selective deletion of the tet fragment but extends the boundaries of the deletion beyond the SnaB I site to at least the BspH I site (see FIG.\",\n \"1).\",\n \"Is the deletion a precise reversion to fd?\",\n \"If so, one would reconstitute the original BamH I site used to clone the tet fragment, and thus generate two restriction fragments (3425 bp and 2989 bp as in the fd BamH I digest, see FIG.\",\n \"4A lane 3) at the expense of the linearized deletion product (see for example the Msc I digest in FIG.\",\n \"2B lane 2).\",\n \"As can be seen in FIG.\",\n \"4A (lane 2), BamH I digestion of fd-tet isolated from K91KAN produced a pattern similar to that shown for Msc I digest.\",\n \"Two bands corresponding to the linearized fd-tet (9183 bp) and the full-length linearized deletion product (approximately 6.4 kb) were generated.\",\n \"This thus illustrates that although the deletion encompasses much or all of the tet fragment, it does not precisely regenerate the second BamH I site of the fd parent molecule.\",\n \"Thus, the next question addressed, once the 2.8 kbp deletion was mapped to the vicinity of the tet fragment, was whether or not the excision sites lay within or outside of the tet fragment.\",\n \"For this the BstY I digest illustrated in FIG.\",\n \"4B was performed (two BstY I sites, created during the fd-tet construction, precisely bracket the tet fragment, see FIG.\",\n \"1).\",\n \"The gel pattern obtained for the BstY I digest of fd-tet derived from K802 had three fragments (3425 bp, 2983 bp and 2775 bp) identical in size to those generated when K91KAN derived DNA was used (FIG.\",\n \"4B, compare lane 1 with lane 2).\",\n \"This demonstrates that at least one of the two BstY I sites was retained in the deleted product.\",\n \"Therefore, sequences immediately adjacent to the original BamH I site, must also be preserved in the deleted fd-tet, a fact that may be relevant to the mechanism of the deletion process (see Discussion).\",\n \"The Deletion is recA Independent and F+Dependent K802 and K91KAN strains of E. coli were introduced by Smith to produce and use the phage display vectors he has pioneered (Parmley, 1988).\",\n \"The principle consideration is that phage infectability is dependent on the F-factor.\",\n \"Therefore, manipulation of the DNA vector, its modifications and construction can be made in the absence of infection in F-bacteria (such as K802), thus avoiding multiply infected bacteria.\",\n \"Once the vector is completed, then in order to screen a phage library or to amplify and generate high titers of phages one must use F+ bacteria (e.g., K91KAN).\",\n \"Table 1 illustrates that in a variety of E. coli strains tested, the tet deletions were detected only in the F+ strains.\",\n \"Most noteworthy is the comparison between the isogenic DH5α and DH5α F′ E. coli strains.\",\n \"Here too the deletion occurs exclusively in the DH5αF′ strain.\",\n \"These strains are identical in their genotypes except that DH5αF′ carries the F-factor.\",\n \"This indicates that the deletion is F-factor dependent.\",\n \"Moreover, both strains are RecA− in which the recA gene is mutated and thus classical homologous recombination is not possible.\",\n \"Therefore, the deletion event is recA independent.\",\n \"TABLE 1 The Deletion is F+ Dependent and recA Independent E. coli Strain redA F-episome Deletion K802 + − − K91KAN + + + DH5αa − − − DH5αF′a − + + JM109 − + + XL1-Blue − + + MC1061b + − − NM554b − − − aDH5α and DH5αF′ are isogenic strains; DH5αF′ contains the F-episome bMC1061 and NM554 are isogenic strains; NM554 is recA− As the F-factor is essential when using filamentous phages, working with F+ bacterial strains cannot be avoided.\",\n \"Therefore, introducing the recombinant pVIII gene in a region prone to F+ dependent deletions is potentially problematic.\",\n \"The suitability of the non-coding region between the wild type pVIII gene and the pIII gene as a site for genetic manipulation and introduction of a second pVIII gene was therefore explored.\",\n \"Construction of the ftac88 Vector Designing a phage display vector with the aim of introducing a second pVIII gene in the non-coding region between the wild type pVIII and pIII genes presents two obstacles that must be considered.\",\n \"1.Unique restriction sites for cloning are unavailable in this region.\",\n \"2.This region contains an essential overlapping promoter and rho-independent transcription terminator that are inseparable (see FIG.\",\n \"5; see also Van Wezenbeek et al, 1980).\",\n \"The terminator terminates the transcription of upstream genes ending with the pVIII gene, whereas the promoter initiates the transcription of downstream genes starting with the pIII gene.\",\n \"Therefore, the strategy adopted to insert the recombinant second pVIII gene was to duplicate this promoter/terminator as well.\",\n \"Ultimately, a “type 88” expression vector (designated ftac88) was constructed as follows: The first step was to generate the modified pVIII gene.\",\n \"This was designed to contain unique restriction sites that could be used to introduce foreign DNA sequences and thus enable the presentation of their corresponding encoded peptides at the N-terminus of the pVIII protein.\",\n \"Such a modified pVIII gene, designated pVIIISTS, was produced by “SOEing” PCR (Horton et al, 1990) mutagenesis in which the GAC codon for residue Asp, at position 4 of mature wild type pVIII protein, was replaced by an insert of 62 base pairs (“STS” insert, FIG.\",\n \"6A).\",\n \"This insert contained two Sfi I sites that bracketed two translation stop-codons followed by the trpA transcription terminator.\",\n \"Thus, the transcription of the pVIIISTS gene terminates prematurely, rendering the expression of this gene silent.\",\n \"In order to express this pVIII gene one must exchange the Sfi I flanked stuffer for a functional in-frame insert of DNA.\",\n \"The PCR was designed to generate a fragment corresponding to the SnaB I-BamH I segment of fd-tet (see FIG.\",\n \"5) that was cloned into the pGEM-T vector, designated pGEM-T(p8STS) (FIG.\",\n \"6B).\",\n \"Next, a wild type pVIII gene was cloned into the SnaB I site of pGEM-T(p8STS).\",\n \"This was achieved by PCR of the wild type pVIII gene segment from fd-tet starting from the upstream SnaB I site and continuing just beyond the overlapping promoter/terminator.\",\n \"The downstream antisense primer incorporated three unique restriction sites as well.\",\n \"The PCR product was cloned into the blunt ends of SnaB I cut pGEM-T(p8STS) thus generating the desired pGEM-T(p88STS).\",\n \"As is illustrated in FIG.\",\n \"6C the SnaB I-BamH I fragment of pGEM-T(p88STS) contains two pVIII genes, duplicated promoter/terminator elements and several unique cloning sites.\",\n \"To complete the generation of the “type 88” phage display vector all that was required was to replace the SnaB I-BamH I segment of fd-tet with the corresponding SnaB I-BamH I fragment of pGEM-T(p88STS).\",\n \"However, fd-tet contains two SnaB I sites (see FIG.\",\n \"5).\",\n \"Therefore, the SnaB I site in the tetR gene of fd-tet was removed.\",\n \"For this, fd-tet genome was digested with Xba I/BstX I to remove the 725 bp fragment containing the SnaB I site that was replaced by a linker containing an Ava II site.\",\n \"This intermediate modified fd-tet was then digested with SnaB I/BamH I and the wild type segment was exchanged for the correspondingly cut fragment derived from pGEM-T(p88STS).\",\n \"Furthermore, a tac promoter was introduced between the Xho I and Sac I sites to produce the “type 88” vector designated ftac88 shown in FIG.\",\n \"7.It was then necessary to evaluate whether or not the ftac88 vector can produce mosaic phages displaying both wild type and chimeric pVIII proteins.\",\n \"For this a linker encoding the linear peptide APAGFAIL (SEQ ID NO:28) (the epitope corresponding to the anti-gp120 mAb GV4H3 (Denisova et al, 1996)) was inserted in frame between the two Sfi I sites, eliminating the trpA terminator.\",\n \"As demonstrated by the dot blot analysis (FIG.\",\n \"8), this construct (ftac88(4H3)) produced phages that were recognized by the GV4H3 mAb.\",\n \"Genetic Instability in ftac88 The orientation of the wild type pVIII and the modified pVIIISTS genes in the ftac88 vector is a direct tandem repeat.\",\n \"The construction of this vector was performed in recA− bacteria and as such excludes the possibility of recA dependent homologous recombination.\",\n \"However, in view of the fact that the deletion of the tet fragment in fd-tet (described above) is recA independent and F+ dependent, the genetic stability of the modified pVIIISTS gene in DH5αF′ was examined.\",\n \"For this, ftac88 DNA preparations from DH5α and DH5αF′ bacteria were compared.\",\n \"Double digestion of ftac88 with SnaB I and BamH I should release a 1378 bp fragment containing both pVIII genes (see FIG.\",\n \"7).\",\n \"Indeed, SnaB I/BamH I double digestion produced only the expected 1378 bp fragment, regardless of the source of the DNA (FIG.\",\n \"9, lane 1, 2).\",\n \"As expected, the deletion of the tet fragment still occurred in DH5αF′ (FIG.\",\n \"9, lane 2; indicated by the upper arrow).\",\n \"Thus one can conclude that the modified pVIIISTS gene is genetically stable in ftac88.However, the purpose of the ftac88 vector is to display peptides as chimeric pVIII proteins and as such, it was important to further investigate the genetic stability of the modified pVIIISTS gene when it contained a peptide coding sequence rather than the stuffer with the trpA terminator.\",\n \"For this, the ftac88 DNA construct expressing the GV4H3 epitope (above) was prepared from DH5α and DH5αF′ bacteria and used for the double digestion, SnaB I/BamH I analysis, as above.\",\n \"Here the situation was surprising.\",\n \"Digestion of the DNA derived from DH5α released as expected a 1376 bp fragment (FIG.\",\n \"9, lane 3).\",\n \"However, SnaB I/BamH I digestion of the same DNA prepared from DH5αF′ generated an additional smaller fragment of approximately 950 bp (FIG.\",\n \"9, lane 4).\",\n \"This indicates that a deletion of approximately 400 bp occurred.\",\n \"This deletion corresponded to selective loss of the recombinant pVIII gene and reconstitution of the wild type pVIII-pIII configuration as confirmed by sequence analysis (not shown).\",\n \"Therefore, it was concluded that the modified pVIIISTS gene, when containing coding sequences, becomes genetically unstable in DH5αF′.\",\n \"This also suggests that the stability of the ftac88 observed in DH5αF′ is rendered due to the trpA terminator which could imply that the deletion process may require transcribed regions.\",\n \"In summary, it is clear that the non-coding region between pVIII and pIII can be used.\",\n \"Moreover, the obstacle of the overlapping promoter/terminator can be overcome.\",\n \"However, genetic instability of the recombinant pVIII gene was still a problem.\",\n \"Construction of fth1 Vector In light of the above experience in designing, constructing and testing the ftac88 vector, it was decided to redesign a novel “type 88” vector that would overcome the problems of genetic instability and include other improvements.\",\n \"The attributes of such a vector, designated fth1 (FIG.\",\n \"10) are listed herewith: 3.The second pVIII gene contained the 62 bp “STS” insert as in ftac88.However, the gene was constructed using synthetic oligonucleotides to generate a pVIII protein of identical amino acid sequence yet coded for with alternative codon usage.\",\n \"This obviated the formation of tandem repeats, a configuration that is prone to deletion events as illustrated in the analysis of ftac88 (above).\",\n \"The homologous gene was designated pVIIISTSh.\",\n \"The pVIIISTSh gene was cloned, as in ftac88, between the wild type pVIII and pIII genes 4.To avoid the generation of additional tandem repeats, an alternative HP terminator (Nohno et al, 1986) was introduced into the 3′ region of the wild type pVIII gene ten codons upstream to the pVIII stop codon.\",\n \"Moreover, the severed 3′ region coding for the C-terminal domain of the wild type pVIII protein was reconstituted using alternative codon usage (so to reduce homology within the −35 box of the promoter of the pIII gene residing in this region (Van Wezenbeek et al, 1980)).\",\n \"5.The tetracycline resistance gene was cloned between the two pVIII genes.\",\n \"This was accomplished via PCR of the open reading frame of the tetracycline resistance gene only and placing it downstream to a kanamycin promoter (derived from M13K07 phage).\",\n \"This was done so as to avoid using the bi-directional tetracycline promoter (see FIG.\",\n \"5) which would otherwise generate an anti-sense transcript of the wild type pVIII gene were it used in this new position.\",\n \"Also the exclusion of the tetracycline repressor is advantageous as the tetracycline resistance gene thus becomes constitutively expressed, making pre-incubation with low levels of tetracycline to induce resistance unnecessary.\",\n \"6.The original intergenic space containing the ori sequences was reconstituted.\",\n \"Previously, the tet fragment used by Smith to construct fd-tet, was inserted into the fd ori(−) rendering this vector and its derivatives low copy number and thus producing relatively low phage titers (Smith, 1988).\",\n \"Reconstituting the intergenic region in fth1 provided two advantages: the copy number of the DNA was greatly increased and the titers of secreted phage increased as well (see below).\",\n \"The construction of fth1 (see FIG.\",\n \"11) was accomplished using “SOEing” PCR as described above enabling the introduction of multiple cloning sites and novel genes into the non-coding region between the wild type pVIII and pIII genes of fd-tet.\",\n \"The first step was to modify fd-tet by the introduction of a short segment of DNA, ten codons upstream from the stop codon of the wild type pVIII gene (FIG.\",\n \"11B).\",\n \"This segment not only provided novel restriction sites but also separated the promoter/terminator adjacent to the pIII gene from the wild type pVIII gene by reconstituting the severed last ten codons of the pVIII open reading frame (intermediate vector 1 (IV-1) in FIG.\",\n \"11B).\",\n \"Subsequently, additional intermediate vectors were produced (IV-2 and IV-3, see Materials and Methods) which eventually led to IV-4 (FIG.\",\n \"1C).\",\n \"In the latter, the novel HP terminator was incorporated downstream to the reconstituted wild type pVIII gene followed by the tetracycline resistance open reading frame driven by the Kanamycin promoter.\",\n \"Moreover, the ori(−) in the intergenic region was also reconstituted in IV-4.The final vector was produced by cloning the synthetic pVIIISTSh gene, driven by the tac promoter, into the Bgl II/Eag I digested IV-4 vector to give fth1 (see detailed map, FIG.\",\n \"10).\",\n \"The questions to be evaluated regarding the utility and advantages of fth1 therefore were: 1.Was the vector, and particularly the tetracycline resistance and recombinant pVIII genes, genetically stable?\",\n \"2.Could the recombinant pVIII gene be used to stably express peptide inserts?\",\n \"3.Was the fth1 a high copy number vector?\",\n \"4.Could high titers of fth1 progeny be achieved?\",\n \"In order to answer these questions, the GV4H3 epitope was introduced between the Sfi I sites in fth1 (as done previously for ftac88).\",\n \"This construct, called fth1(4H3), produced mosaic phages presenting the GV4H3 epitope that were recognized by the GV4H3 mAb (FIG.\",\n \"8).\",\n \"The stability of the tetracycline resistance gene and the modified pVIIISTSh gene was tested using restriction analyses as before.\",\n \"Digestion of fth1 and fth1(4H3) DNA, derived from DH5α or DH5αF′, with BstN I generated identical fragments as is illustrated in FIG.\",\n \"12.BstN I sites appear twice in both fd (see FIG.\",\n \"1) and fth1 (see FIG.\",\n \"10).\",\n \"BstN I digestion of fd generated the expected two fragments of 5456 bp and 952 bp sizes (FIG.\",\n \"12, lane 1).\",\n \"The latter contains the non-coding region between pVIII and pIII genes.\",\n \"As this region in fth1 contains the tetracycline resistance and the recombinant pVIIISTSh genes, the 952 bp fragment was shifted to generate the 2777 bp and 2775 bp bands in BstN I digests of fth1 and fth1(4H3) respectively derived from both DH5α and DH5αF′ (lane 2-5).\",\n \"Any deletions of either the tetracycline resistance gene or the recombinant pVIII gene or their parts would generate DNA fragments ranging in size from 952 bp to 2777 bp.\",\n \"No such fragments could be detected illustrating that the fth1 vector is genetically stable.\",\n \"The functionality of the reconstituted ori(−) is directly apparent.\",\n \"In several independent purifications of fth1 DNA, 0.5-1 mg DNA/liter of bacteria was routinely obtained, as opposed to the typical yields of 50 μg DNA per liter of ftac88 harboring bacteria.\",\n \"Similarly a hundred fold increase is typically seen in phage titers for which 1012 phages/ml is measured for fth1 as compared to 1010 phages/ml achieved for fd-tet and its derivatives.\",\n \"Thus the fth1 vector provides a convenient and stable “type 88” phage display expression system.\",\n \"Discussion The selective deletion of the tet fragment from the intergenic space has been critically evaluated as a first step towards producing a novel “type 88” phage display vector.\",\n \"The present inventors have observed that in F+ bacterial strains a 2.8 kb fragment of DNA is excised from the region of the foreign tet insert, Smith originally introduced into the B-stem/loop of the ori(−) structure (Zacher et al, 1980).\",\n \"The loss of the insert's Ava I site, which overlaps the original remnant BamH I site, illustrates that downstream aspects of the insert are not preserved in the deleted form.\",\n \"The loss of the BspH I site and all four of the insert's Hinc II sites (not shown) argues that no more than 18 base pairs of the insert can be retained at the other extreme.\",\n \"Preservation of at least one BstY I site, yet lack of reconstitution of the original BamH I site, further stresses the close but not precise correspondence of the deleted fragment with the original tet insert.\",\n \"As the insert is flanked by the inverted repeats that create the B-stem/loop and that these repeats are retained in the deleted form of the phage, one might postulate that the mechanism generating the deletion involves homology and base paring.\",\n \"Clearly, however, this can not be homologous recombination as that would require direct repeats of homologous stretches of over 50 bases and of course, a functional RecA protein in order to enable a deletion event (Clark, 1973; Matfield et al, 1985).\",\n \"The deletion events in our experiments occurred in recA− bacterial strains.\",\n \"An alternative mechanism, the “slippage mechanism” proposed by Lovett (Feschenko et al, 1998), also requires direct repeats and therefore can not explain the deletion.\",\n \"To the best of our knowledge, no known mechanism can explain the deletion of the tet fragment as observed.\",\n \"One might imagine that during the course of replication, the inverted repeats, flanking the tet fragment, align to form a B-stem/loop of sorts, greatly distorted by a grossly extended “bubble” (where the BamH I site had been), consisting of the 2.8 kb insert.\",\n \"During replication, the polymerase might be able to “skip” from one side of the bubble to the other thus omitting the entire insert and generating the deletion.\",\n \"Assuming that this is a very rare event one must still address the fact that in K91Kan cells over 50% of the DNA isolated was of the deleted form.\",\n \"Moreover, why is the deletion F+ dependent?\",\n \"The matter of copy number might be explained due to two selective advantages that are realized in the deleted form.\",\n \"The mere fact that the genome is 2.8 kb shorter than fd-tet, should speed up the replication.\",\n \"More important may be the fact that the deletion most probably partially reconstitutes the ori(−) structure.\",\n \"The ramifications of a markedly more functional ori(−) would substantially enable more effective production of double stranded DNA.\",\n \"Another possibility may be related to the packaging of phages for secretion.\",\n \"The A-stem/loop structure, situated adjacent to the B-stem/loop, is the packaging-signal of the phage (Van Wezenbeek et al, 1980).\",\n \"It stands to reason that the tet insert may sterically hinder the packaging of the recombinant phages, a burden that would be removed in the deleted form.\",\n \"However, the major phenomenon encountered here is the fact that the deletion is detectable only in the F+ bacteria.\",\n \"This might be due to the fact that even rare events can be markedly amplified when super infection of bacteria can take place.\",\n \"Obviously, for such super infection, the bacteria must be compatible, a situation only satisfied in F+ bacteria.\",\n \"Therefore, it is proposed that the deletion leads to shorter and more efficient replicating forms of the phage, albeit a rare event.\",\n \"Subsequently, however, such deleted phages can be markedly amplified in F+ bacteria via super infection thus enabling accumulation of very substantial amounts of the deleted form.\",\n \"However, the possibility cannot be excluded that unknown proteins encoded by the F-episome might contribute to the deletion events either by directly enhancing the process or indirectly inducing the expression of bacterial proteins participating in the process.\",\n \"In view of the above considerations, it was concluded that the intergenic region of the phage is not the optimal site for introduction of a second pVIII gene.\",\n \"Not that this has not led to an effective “type 88” vector in the past such as the f88.4 vector constructed by Smith (Zhong et al, 1994).\",\n \"In the f88.4 vector, the recombinant pVIII gene was introduced into the tet fragment of fd-tet.\",\n \"However, one would expect with such a vector the gradual appearance of deleted forms of the phages and loss of their peptide inserts displayed by the chimeric pVIII protein.\",\n \"Indeed, such “contaminating”, fast-growing, tetracycline sensitive phages have been reported by Scott when they used the f88.4 vector (Bonnycastle et al, 1996).\",\n \"An alternative cloning site for the second pVIII gene was tested, namely the non-coding region between the pVIII and pIII genes.\",\n \"The ftac88 described in this report illustrated that such a construction is not only possible but able to produce workable amounts of recombinant phages.\",\n \"However, once again an unexpected obstacle was encountered as substantial amounts of deletions of the recombinant pVIII were observed.\",\n \"This deletion process also occurred in a recA− background, excluding therefore the possibility of classical homologous recombination.\",\n \"In contrast to the situation of the deletion of the tet insert however, the deletion of the modified pVIII gene entails reasonably long direct repeats.\",\n \"Thus, whereas homologous recombination is not an option, the slippage mechanism which is recA independent, described by Feschenko and Lovett (Feschenko et al, 1998), might be responsible for the selective loss of the second pVIII gene in ftac88.This would entail the slipped misalignment of the nascent strand during DNA replication.\",\n \"During replication of the repeated DNA the nascent strand becomes displaced from its template and pairs with the other copy on the template strand.\",\n \"Then, replication is resumed and the slipped misalignment can lead to either deletion or expansion within tandem repeat arrays.\",\n \"Therefore, deletions due to the slippage mechanism are homology dependent, a requirement satisfied in ftac88.The selective loss of the recombinant pVIII gene further supports the slippage mechanism.\",\n \"This mechanism requires the stabilization of the slipped misalignment structure (Feschenko et al, 1998; Lovett et al, 1996).\",\n \"The longer the displaced nascent strand, the more stabilized the slipped misalignment structure becomes.\",\n \"The recombinant pVIII gene contains a foreign sequence in its 5′ region.\",\n \"This severs the repeat into two parts, a short sequence upstream and a long downstream sequence.\",\n \"The long sequence, therefore, enables the stabilization of the slipped misaligned structure more efficiently than the short one.\",\n \"Therefore, excision sites in the long downstream identical sequence should predominate thus eliminating upstream sequences including the foreign DNA insert.\",\n \"The actual deletion event may be a very rare one, typical of the slippage mechanism.\",\n \"One can postulate that deleted DNA products could be amplified due to F+ dependent superinfection (see discussion above).\",\n \"Therefore, the last aspect that must be considered is why the pVIIISTS form is stable as opposed to those constructs in which the stuffer containing the trpA terminator is exchanged for a peptide coding-sequence (such as the GV4H3 epitope).\",\n \"The stability rendered by the presence of the trpA transcriptional terminator indicates that transcription might be a positive and necessary factor.\",\n \"The trpA terminator stops the transcription of the recombinant pVIII gene prematurely and consequently no deletion is observed.\",\n \"Transcription through the second homologous DNA repeat appears to be a necessary pre-requisite that provides the opportunity to form the misaligned structure that leads to subsequent deletion during DNA replication.\",\n \"Such a mechanism may explain how misalignment can exist between the first and second repeats at the moment when the first repeat is being replicated yet the second downstream repeat is still in the closed double stranded DNA configuration.\",\n \"In order to mispair the nascent strand to the closed double-stranded repeat, the latter would have to be opened.\",\n \"In the absence of RecA protein, which catalyzes such reactions (West, 1992), transcription might be an efficient mode for opening double stranded DNA.\",\n \"However, as opening double stranded DNA during transcription is transient, this mode would be efficient only when the DNA repeats are in reasonably close proximity.\",\n \"Indeed, the deletion frequency, reported for the slippage mechanism, is correlated directly with the distance separating the DNA repeats (Lovett et al, 1994; Bi et al, 1994).\",\n \"Moreover, this mechanism would require that the wild type pVIII gene be the first to be replicated.\",\n \"Indeed, the most extensive replication during infection is that of the + strand, proceeding from the intergenic region through the wild type pVIII gene to the recombinant pVIII gene (Horiuchi et al, 1976).\",\n \"As a result of the above analyses and the experiences gained through the construction and characterization of the ftac88 vector, an improved novel vector that corrects for the problems encountered was ultimately designed.\",\n \"This vector has been designated fth1 and has the following advantages.\",\n \"1.The tetracycline resistance and the recombinant pVIII genes were introduced between the wild type pVIII and pIII genes.\",\n \"2.To avoid the presence of repetitive sequences of any kind, the two transcription units of the phage genes were separated by inserting a novel HP-terminator.\",\n \"Furthermore, the wild type pVIII C-terminal domain encoding region was reconstituted using alternative codon usage.\",\n \"The recombinant pVIII gene was generated using alternative codon usage as well.\",\n \"These attributes render the fth1 vector genetically stable.\",\n \"3.The fth1 vector contains the wild type intergenic region.\",\n \"Thus, the minus-strand DNA synthesis occurs as efficiently as in wild type fd rendering fth1 a high copy number vector.\",\n \"This enables one to isolate and purify consistently large amounts of the double-stranded DNA from fth1 harboring bacteria (500 μg per liter as oppose to 50 μg of ftac88).\",\n \"Furthermore, fth1 produces high phage titers −1012 phages per ml as oppose to 1010 phages per ml typical for fd-tet and its derivatives.\",\n \"The foregoing description of the specific embodiments will so fully reveal the general nature of the invention that others can, by applying current knowledge, readily modify and/or adapt for various applications such specific embodiments without undue experimentation and without departing from the generic concept, and, therefore, such adaptations and modifications should and are intended to be comprehended within the meaning and range of equivalents of the disclosed embodiments.\",\n \"It is to be understood that the phraseology or terminology employed herein is for the purpose of description and not of limitation.\",\n \"The means, materials, and steps for carrying out various disclosed functions may take a variety of alternative forms without departing from the invention.\",\n \"Thus the expressions “means to .\",\n \".\",\n \".\",\n \"” and “means for .\",\n \".\",\n \".\",\n \"”, or any method step language, as may be found in the specification above and/or in the claims below, followed by a functional statement, are intended to define and cover whatever structural, physical, chemical or electrical element or structure, or whatever method step, which may now or in the future exist which carries out the recited function, whether or not precisely equivalent to the embodiment or embodiments disclosed in the specification above, i.e., other means or steps for carrying out the same functions can be used; and it is intended that such expressions be given their broadest interpretation.\",\n \"REFERENCES Bi et al, “recA-independent and 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Cabilly (ed.\",\n \"), Humana Press, Totowa, N.J., p. 137 (1998) Sugimoto et al, “Studies on bacteriophage fd DNA.\",\n \"IV.\",\n \"The sequence of messenger RNA for the major coat protein gene”, J Mol Biol 111:487-507 (1977) Van Wezenbeek et al, “Nucleotide sequence of the filamentous bacteriophage M13 DNA genome: comparison with phage fd”, Gene 11:129-148 (1980) Vieira et al, “Production of single-stranded plasmid DNA”, Methods Enzymol 153:3-11 (1987) West S C, “Enzymes and molecular mechanisms of genetic recombination”, Annu Rev Biochem 61:603-640 (1992) Willis et al, “Immunological properties of foreign peptides in multiple display on a filamentous bacteriophage”, Gene 128:79-83 (1993) Wrighton et al, “Small peptides as potent mimetics of the protein hormone erythropoietin”, Science 273:458-463 (1996) Zacher et al, “A new filamentous phage cloning vector: fd-tet”, Gene 9:127-140 (1980) Zhong et al, “Conformational mimicry of a chlamydial neutralization epitope on filamentous phage”, J Biol Chem 269:24183-24188 (1994\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10474148"}}},{"rowIdx":784,"cells":{"text":{"kind":"list like","value":[["Enantiomerically selective cyclopropanation","A method of forming a cyclopropane having enhanced chirality said method comprising reacting together: a symmetrical 1,2-dioxine of the formula (1), wherein X and Y are the same and are groups in which a carbon atom is bonded to the dioxine backbone; and a phosphorus ylide or a phosphorus ylide precursor, in the presence of a cobalt catalyst containing a chiral ligand."],["1.A method of forming a cyclopropane having enhanced chirality said method comprising reacting together: i) a symmetrical 1,2-dioxine of the formula (1), wherein X and Y are the same and are groups in which a carbon atom is bonded to the dioxine backbone; and ii) a phosphorus ylide or a phosphorus ylide precursor; in the presence of iii) a cobalt catalyst containing a chiral ligand.","2.A method of forming a cyclopropane having enhanced chirality according to claim 1, wherein the chiral ligand in the catalyst has the structure: —(OR′C═R′CR′C═NR′CHCHR′N═CR′CR′═COR′)— around the cobalt atom and wherein each R′ moiety may be independently selected from a wide range of groups including, but not limited to H, alkyl, aryl, keto, ester and a range of other functionalities.","3.A method of forming a cyclopropane having enhanced chirality according to claim 1, in which the cobalt catalyst has the structure: Wherein R1 and R2 may be hydrogen, alkyl, aryl; R3 may be hydrogen, alkyl, t-alkyl, alkyl or aryl, alkoxy, aryl alkoxy; R3, R4, R5 and R6 may be independently H, alky, aryl, keto, or ester.","4.A method of forming a cyclopropane having enhanced chirality according to claim 3, in which R1 and R2 are each the same and are selected from hydrogen, (CH2)4, or benzyl.","5.A method of forming a cyclopropane having enhanced chirality according to claim 3, in which R3 is hydrogen, t-butyl, or benzyl ethyl, bornoxy, menthoxy, ethoxy or methyl.","6.A method of forming a cyclopropane having enhanced chirality according to claim 3, in which R4 and R5 may be the same or different and are selected from hydrogen, and t-butyl.","7.A method of forming a cyclopropane having enhanced chirality according to claim 1 in which the phosphorus ylide or phosphorus ylide precursor may be taken to mean a stabilized phosphorus ylide or a reactive compound capable of forming a phosphorus ylide in situ.","8.A method of forming a cyclopropane having enhanced chirality according to claim 1, in which the phosphorus ylide or phosphorus ylide precursor is a compound of the formula: i) R7R8R9P=CZCO2R10; wherein R7, R8 and R9 may be the same or different and may be alkyl, substituted alkyl, aryl, substituted aryl; R10 represents a non bulky group such ac C1-4 alkyl or substituted alky, or a bulky group such as 1-adamantyl; and Z represents hydrogen or methyl; or ii) R7R8R9P═CONR10R11; wherein R7, R8, R9 and R10 have the values identified in (i) above and wherein R11 represents a C1-12 alkoxy grouping; iii) R7R8R9P═CN; or iv) (R70)2P(O)CH2CO2 R10 wherein R7 and R10 have the values identified in (i) above.","9.A method of forming a cyclopropane having enhanced chirality according to claim 1, in which the phosphorus ylide is a non-bulky stabilized ylide.","10.A method of forming a cyclopropane having enhanced chirality according to claim 9, in which the non-bulky stabilized ylide is benzyl 2-(triphenyl-λ-5 phosphanylidene) acetate.","11.A method of forming a cyclopropane having enhanced chirality according to claim 1, in which the phosphorus ylide is a bulky ylide.","12.A method of forming a cyclopropane having enhanced chirality according to claim 1, in which the bulky ylide is t-butyl 2-(triphenyl-λ-phosphanylidene) acetate.","13.A method of forming a cyclopropane having enhanced chirality according to claim 1, in which the phosphorus ylide or phosphorus ylide precursor is selected from N,N-methoxymethyl-2-(1,1,1-triphenyl-λ5-phosphanylidene)acetamide; 2-(1,1,1-triphenyl-λ5-phosphanylidene) acetonitrile or methyl 2-(dimethoxyphosphoryl) acetate.","14.A method of forming a cyclopropane having enhanced chirality according to claim 1, in which X and Y are selected from H, alkyl or aryl groups.","15.A method of forming a cyclopropane having enhanced chirality according to claim 1, wherein the reaction is conducted in a solvent selected from acetonitrile, ethyl acetate/hexane admixtures, ethyl acetate, tetrahydrofuran, ether, toluene, acetone, carbon tetrachloride, and dichloromethane or mixtures thereof.","16.A method of forming a cyclopropane having enhanced chirality according to claim 1, in which the reaction is conducted with catalyst concentration of up to 50 mol % catalyst relative to the 1,2-dioxine, and more preferably a catalyst concentration of 1-15 mol %.","17.A method of forming a cyclopropane having enhanced chirality according to claim 1, wherein the optically enriched cyclopropanes are subjected to recrystallization or column chromatography.","18.A method of forming a cyclopropane having enhanced chirality according to claim 1, in which the enhanced chirality of the product, as determined by chiral shift n.m.r.","techniques and expressed as an enantiomeric ratio may be as high as 90/10."],["<SOH> BACKGROUND OF THE INVENTION <EOH>Cyclopropanes containing natural and non-natural products are receiving considerable attention as synthetic targets since the incorporation of the rigidified cyclopropyl skeleton into bioactive analogues leads to conformationally constrained molecules.","Such modifications often have significant effects on bioactivities with concomitant medical implications.","Cyclopropanes are also generated as transient species in primary and secondary metabolisms in man, plants and microorganisms.","Thus, the great importance of functionalised cyclopropanes in organic synthesis spurs a continuing search for efficient stereo controlled cyclopropanation methodologies.","Of particular importance is the deficiency in methods for the construction of diversely functionalised cyclopropanes that contain greater than di-substitution.","Chemicals containing the cyclopropyl moiety may be used inter alia: in the preparation of cyclopropyl amino acids; the preparation of cyclopropyl pharmaceuticals; the preparation of cyclopropyl pesticides; the preparation of new cyclopropyl products for furrier biological evaluation by chemistry and biochemistry laboratories as well as the preparation of isotopically labelled cyclopropanes as bioactive probes.","Cyclopropanes may be synthesised by a number of techniques including (i) the direct carbene transfer (both stoichiometric and catalytic) from a diazo precursor to an olefin utilizing transition metals (Rh, Cu, Zn, and Pd) and (ii) Michael addition of nucleophiles (usually sulphur ylides) to α, β-unsaturated ketones and esters followed by intra-molecular cyclization.","Such methods, employing as they do, potentially explosive material or requiring reaction mixtures that are highly sensitive to moisture have certain practical drawbacks.","An alternative reaction scheme for the synthesis of cyclopropanes involves the reaction between 1,2-dioxines and stabilised phosphorus ylides, is illustrated in reaction scheme 1.It is suggested that the ylide acts as a mild base inducing the ring opening of the 1,2-dioxine (1) to their isomeric cis γ-hydroxy enones (3) followed by Michael addition of the y]ide to the enone and attachment of the electrophilic phosphorus pole of the ylide to the hydroxyl moiety affording the intermediate 1-2λ-oxaphospholanes (4).","Cyclization of the resultant enolate produces th cyclopropanes (5) in excellent yield.","The cyclopropane products emerging from this reaction scheme have in the past been prepared as racemic mixtures.","The present invention is directed to a synthetic route for the preparation of optically enriched cyclopropane compounds."],["<SOH> SUMMARY OF THE INVENTION <EOH>Therefore according to a first aspect of the invention there is provided a method of forming a cyclopropane having enhanced chirality said method comprising reacting together: (i) a symmetrical 1,2-dioxine of the formula (1), wherein X and Y are the same and are groups in which a carbon atom is bonded to the dioxine backbone; and (ii) a phosphorus ylide or a phosphorus ylide precursor; in the presence of (iii) a cobalt catalyst containing a chiral ligand.","Preferably, the cobalt catalyst hils one of the structures shown below wherein R1 and R2 may be hydrogen, alkyl, aryl; R3 may be hydrogen, alkyl, t-alkyl, alkyl or aryl, alkoxy, aryl alkoxy; R3, R4, R5 and R6 may be independently H, alkyl, aryl, keto, or ester In accordance with a further aspect of the invention there is provided a chiral cyclopropanation catalyst of the formulae shown below in Table 1 based on Cobalt Salen compounds and on Cobalt Beta-Ketoiminato compounds."],["FIELD OF THE INVENTION The present invention relates to a process for introducing an asymmetric cyclopropyl group into a molecule BACKGROUND OF THE INVENTION Cyclopropanes containing natural and non-natural products are receiving considerable attention as synthetic targets since the incorporation of the rigidified cyclopropyl skeleton into bioactive analogues leads to conformationally constrained molecules.","Such modifications often have significant effects on bioactivities with concomitant medical implications.","Cyclopropanes are also generated as transient species in primary and secondary metabolisms in man, plants and microorganisms.","Thus, the great importance of functionalised cyclopropanes in organic synthesis spurs a continuing search for efficient stereo controlled cyclopropanation methodologies.","Of particular importance is the deficiency in methods for the construction of diversely functionalised cyclopropanes that contain greater than di-substitution.","Chemicals containing the cyclopropyl moiety may be used inter alia: in the preparation of cyclopropyl amino acids; the preparation of cyclopropyl pharmaceuticals; the preparation of cyclopropyl pesticides; the preparation of new cyclopropyl products for furrier biological evaluation by chemistry and biochemistry laboratories as well as the preparation of isotopically labelled cyclopropanes as bioactive probes.","Cyclopropanes may be synthesised by a number of techniques including (i) the direct carbene transfer (both stoichiometric and catalytic) from a diazo precursor to an olefin utilizing transition metals (Rh, Cu, Zn, and Pd) and (ii) Michael addition of nucleophiles (usually sulphur ylides) to α, β-unsaturated ketones and esters followed by intra-molecular cyclization.","Such methods, employing as they do, potentially explosive material or requiring reaction mixtures that are highly sensitive to moisture have certain practical drawbacks.","An alternative reaction scheme for the synthesis of cyclopropanes involves the reaction between 1,2-dioxines and stabilised phosphorus ylides, is illustrated in reaction scheme 1.It is suggested that the ylide acts as a mild base inducing the ring opening of the 1,2-dioxine (1) to their isomeric cis γ-hydroxy enones (3) followed by Michael addition of the y]ide to the enone and attachment of the electrophilic phosphorus pole of the ylide to the hydroxyl moiety affording the intermediate 1-2λ-oxaphospholanes (4).","Cyclization of the resultant enolate produces th cyclopropanes (5) in excellent yield.","The cyclopropane products emerging from this reaction scheme have in the past been prepared as racemic mixtures.","The present invention is directed to a synthetic route for the preparation of optically enriched cyclopropane compounds.","SUMMARY OF THE INVENTION Therefore according to a first aspect of the invention there is provided a method of forming a cyclopropane having enhanced chirality said method comprising reacting together: (i) a symmetrical 1,2-dioxine of the formula (1), wherein X and Y are the same and are groups in which a carbon atom is bonded to the dioxine backbone; and (ii) a phosphorus ylide or a phosphorus ylide precursor; in the presence of (iii) a cobalt catalyst containing a chiral ligand.","Preferably, the cobalt catalyst hils one of the structures shown below wherein R1 and R2 may be hydrogen, alkyl, aryl; R3 may be hydrogen, alkyl, t-alkyl, alkyl or aryl, alkoxy, aryl alkoxy; R3, R4, R5 and R6 may be independently H, alkyl, aryl, keto, or ester In accordance with a further aspect of the invention there is provided a chiral cyclopropanation catalyst of the formulae shown below in Table 1 based on Cobalt Salen compounds and on Cobalt Beta-Ketoiminato compounds.","Chiral Cobalt Salen Based Catalysts 8a: R1,R2=—(CH2)4—, R3=t-Bu, R4=H, R5=t-Bu (R,R isomer) 8b: R1,R2=—(CH2)4—, R3=R4=R5=H (R,R isomer) 8c: R1,R2=—(CH2)4—, R3=t-Bu, R4=R5=H (R,R isomer) 8d: R1,R2=Ph, R3=R4=R5=H (R,R isomer) 8e: R1,R2=Ph, R3=t-Bu, R4=R5=H (R,R isomer) 8f: R1,R2=—(CH2)4—, R3=S-PhEtCH—,R4=R5=H (R,R isomer) 8g: R1,R2=Ph, R3=S-PhEtCH—, 14=R5=H (R,R isomer) 8h: R1,R2=Ph, R3=S-PhEtCH—, 14=R5H (S,S isomer) Beta-Ketoiminato Cobalt Based Catalysts 9a: R1=R2=H, R6=(−)-bornoxy 9b: R1=R2=Ph, R6=(−)-bornoxy (S,S isomer) 9c: R1=R2=Ph, R6=(−)-bornoxy (R,R isomer) 9d: R1=R2=—(CH2)4—, R6=(−)-bornoxy(R,R isomer) 9e: R1=R2=Ph, R6=(−)-menthoxy (S,S isomer) 9f: R1=R2=Ph, R6=(−)-menthoxy (R,R isomer) 9g: R1=R2=Ph, R6=(+)-menthoxy (S,S isomer) 9h: R1=R2=Ph, R6=(+)-menthoxy (R,R isomer) 9i: R1=R2=Ph, R6=ethoxy (S,S isomer) 9j: R1=R2=Ph, ethoxy (R,R isomer) 9k: R1=R2=—(CH2)4—, R6=ethoxy (R,R isomer) 9l: R1=R2=Ph, R6=methyl (S,S isomer) Table 1 Catalysts according to the invention in the forms 8a to 8h may be prepared by reaction of Cobalt acetoacetate with the relevant SALEN ligand.","SALEN ligands may be prepared by reaction of (1S, 2S) or (1R, 2R)-1,2-diphenylethylenediamine and the corresponding salicyaldehydes in ethanol under nitrogen atmosphere.","The catalysts 9a-9l may be prepared from reaction of (S,S) or (R,R)-1,2-diphenyl-ethylenediamine and the relevant β-diketone derivative, followed by subsequent reaction with Co acetate.","Corresponding ligands can be made from the (1S, 2S) and (1R, 2R) cyclohexyl-1,2-diamine.","It can be seen that each of the SALEN and the beta-ketoiminato based catalysts has a common structure around the cobalt atom, namely, —(OR′C═R′CR′C═NR′CHCHR′N═CR′CR′═COR′)—.","The catalysts each include at least one chiral grouping.","It is possible for more than one chiral grouping to exist in the catalyst and the values of R′ are wide.","Each of the R′ groups may independently be H, alkyl, aryl, keto, ester and a range of other functionalities.","In the context of the present invention a phosphorus ylide or a phosphorus ylide precursor may be taken to mean a stabilized phosphorus ylide or a reactive compound capable of forming a phosphorus ylide in site.","In a still further aspect of the present invention there is provided a method of forming a cyclopropane having enhanced chirality as described above in which the phosphorus ylide or phosphorus ylide precursor is a compound of the formula: (i) R7R8R9P═CZCO2R10; wherein R7, R8 and R9 may be the same or different and may be alkyl, substituted alkyl, aryl, substituted aryl; R10 represents a non bulky group such as C1-4 alkyl or substituted alkyl, or a bulky group such as 1-adamantyl; and Z represents hydrogen or methyl; or (ii) R7R8R9P═CONR10R11; wherein R7, R8, R9 and R10 have the values identified in (i) above and wherein R11 represents a C1-12 alkoxy grouping; or (iii) R7R8R9P═CN; or (iv) (R7O)2P(O)CH2CO2 R10 wherein R7 and R10 have the values identified in (i) above.","Phosphorus ylides or a phosphorus ylide precursors of type (i) are exemplified by compound (2); type (ii) by compound (10); type (iii) by compound (11); and type (iv) by compound (12).","Preferably, the phosphorus ester ylide is a non-bulky stabilized ylide such as a benzyl 2-(triphenyl-λ-phosphanylidene) acetate (hereinafter referred to as a benzyl ester ylide), or, alternatively, a bulky ylide such as t-butyl 2-(triphenyl-λ-phosphanylidene) acetate (hereinafter referred to as a t-butyl ester ylide).","Alternative phosphorus ylides or ylide precursors include N,N-methoxymethyl-2-(1,1-triphenyl-λ5-phosphanylidene)acetamide; 2-(1,1,1-triphenyl-λ5-phosphanylidene) acetonitrile or methyl 2-(dimethoxyphosphoryl)acetate.","Examples of the reaction between the phosphorus ylides and phosphorus ylide precursors with 1,2-dioxines and a chiral cobalt catalyst and the resulting chiral cyclopropanes are shown below: Preferably, the cyclopropanes of the invention are prepared in a solvent such as acetonitrile, ethyl acetate/hexane admixtures, ethyl acetate, tetrahydrofuran, ether, toluene, acetone, carbon tetrachloride, and dichloromethane or mixtures thereof.","In a particularly preferred form of the invention the cyclopropanes of the invention are prepared in tetrahydrofuran.","It will be appreciated by those skilled in the art that a range of solvents would in fact be suitable for wise in conducting the reaction of the invention.","In any specific case an optimum solvent can be identified by trial and experiment using the above solvents and others.","However, it should also be noted that alcohol is an unsuitable solvent for the reaction.","The enantiomerically enhanced cyclopropanes of the invention are preferably prepared with catalyst concentration of up to 50 mol % catalyst relative to the 1,2-dioxine, and more preferably still a catalyst concentration of 1-15 mol %.","The optically enriched cyclopropanes prepared in accordance with the invention may be further enriched by conventional techniques.","For example, by recrystallization or conversion to diastereoisomers which can be separated by conventional techniques, e.g.","recrystallization or column chromatography.","DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will now be described by way of a number of non limiting examples.","It should be noted that the invention presents a synthetic route for optically enhanced cyclopropanes and that the range of structures capable of synthesis according to the process of the invention is not to be take as being limited to those structures described herein.","An extremely wide range of cyclopropane structures may be manufactured according to the process of the invention.","1.Catalyst Preparation Beta Ketoiminato Catalyst Preparation—The preparation of a chiral cobalt catalyst (9b) (a) Preparation of (−)-Bornyl Acetoacetate To a solution of ethyl acetoacetate (4.5 g, 0.0346 mol) in n-heptane (70 mL) was added (−)-borneol (5.34 g, 0.0346 mol) and sodium hydride (50 mg) and the reaction mixture refluxed under Deans Stark conditions for 2 days.","The solvent was then removed in vacuo and the residue purified by chromatography on silica (1:9 ethyl acetate:hexanes).","(6.16 g, 70%) Rf 0.32 (1:9 ethyl acetate:hexanes) 1H NMR δ 0.85 (s, 3H), 0.88 (s, 3H), 0.91 (s, 3H), 0.94-1.43 (m, 4H) 1.64-2.00 (m, 3H) 2.85 (s, 3H), 3.47 (s, 2H), 4.95 (ddd, 1H, J=15, 5.1, 3.3 Hz).","13C NMR δ 13.35, 18.71, 19.57, 26.96, 27.89, 29.99, 36.51, 44.76, 47.77, 48.76, 50.29, 81.07, 90.10, 167.30.MS m/z 238 (42%, M+), 154 (11%), 137 (100%), 121 (10%), 95 (23%).","(b)(−) Bornyl 2-Formyl-3-Oxobutanoate To the (−)-bornyl acetoacetate (2.0 g, 8.4×10−3 moles) was added N,N-dimethylformamide dimethyl acetal (2 g, 1.68×10−2 moles), the mixture stirred at room temperature for 2 hours, then cooled to 0° C. and methanolic sodium hydroxide (1N, 14 mL 1:l methanol:water) added.","The reaction mixture was stirred for a further 2 hours then cooled to 0° C. and hydrochloric acid (1N) added till pH 3-4.The mixture was extracted with diethyl ether, dried (Na2SO4) and the solvent removed in vacuo.","Rf 0.6 (1:9 ethyl acetate:hexanes) Due to the instability of the aldehyde it was used directly in the next reaction to form the ligand.","1H NMR δ 0.875 (s, 3H), 0.90 (s, 3H), 0.94 (s, 3H), 1.00-1.48 (m, 1H), 1.12-1.48 (m, 4H), 2.32-2.54 (m, 1H), 2.57 (s, 3H), 4.96-5.05 (m, 1H), 9.25 (d, 1H, J-9 Hz).","The crude (−)-bornyl 2-formyl-3-oxobutanoate (0.25 g, 9.49×10−4 mol) was combined in ethanol (3 mL) with (1S, 2S)-1,2-diphenylethylenediamine (0.10 g, 4.75×10−4 mol) and allowed to react for 2 days, the solvent was then removed in vacuo and the residue purified by chromatography (1:9 acetone:dichloromethane) (290 mg, 86%).","Rf 0.64 (1:9 acetone:dichloromethane).","1H NMR δ0.79 (s, 6H), 0.86 (s, 6H), 0.90 (s, 6H), 0.96-1.32 (m, 4H), 1.60-1.82 (m, 6H), 2.28-2.49 (m, 2H), 2.51 (s, 6H), 4.67 (d, 2H, J=7.8 Hz), 4.89 (dm, 2H, J=8.2 Hz), 7.10-7.15 (m, 4H), 7.27-7.33 (m, 6H), 7.79 (d, 2H, J=12.8 Hz), 11.93-12.02 (m, 2H).","13C NMR δ 13.63, 18.84, 19.70, 27.64, 28.03, 31.02, 36.92, 44.84, 47.74, 48.74, 69.53, 79.48, 102.05, 135.68, 158.96, 167.09, 199.71.MS m/z 710 (M+, 4%), 555 (22%), 147 (19%), 354 (14%), 200 (31%), 137 (33%), 95 (100%).","(c) Preparation of Catalyst 9b) A mixture of ligand (144 mg, 2.03×10−4 mol) was combined with cobalt acetate tetrahydrate (50.6 mg, 2.03×10−4) in deaerated ethanol (4 mL) and heated under reflux for 4 hours.","The solvent was then removed in vacuo till dryness of the complex resulted.","The complex was used without further purification.","Each of the catalysts 9a-9l may be prepared using similar methodology and by substitution of the appropriate bornoxy, ethoxy, methoxy, ethoxy or methyl starting materials.","2.Cyclopropane Preparation Using Chiral Beta Ketoiminato Based Catalyst (9a-9l) (a) Preparation of Benzyl 2-(2-Benzoyl-3Phenylcyclopropyl) Acetate 3,6-Diphenyl-3,6dihydro-1,2 dioxine (1a) was reacted with the benzyl 2-(triphenyl-λ5-phosphanylidene)acetate (referred to as benzyl ester ylide) of in the presence of Cobalt (II) catalysts of type 9a-9l with as depicted in scheme 2 to produce the cyclopropane product.","The cobalt catalyst 9b (4.8 mg, 6.3×10−6 mol, 5 mol %) was dissolved in dichloromethane (1 mL) and allowed to equilibrate at the temperature of the reaction.","3,6-Diphenyl-3,6-dihydro-1,2-dioxine (20 mg, 8.4×10−5 mol) was added and the reaction left until such time as complete rearrangement of the dioxine had occurred (t.l.c.).","Benzyl ester ylide (40 mg, 9.8×10−5 mol) was then added and the reaction mixture left for 10 hours.","The solvent was then removed and the residue purified by column chromatography (SiO2, 3:17 ethyl acetate:hexanes).","It is to be noted that the scope of the, invention is not limited to the specific 1,2-dioxines listed in scheme 3.A much greater range of substrates may be used to form a wide variety of substituted cyclopropane products.","It should however be noted that the 1,2-dioxine must be a symmetrical dioxine.","(b) Determination of Enantiomeric Excess Utilising the cyclopropane synthesised in 2(a) above, (5 mg) was dissolved in 1:4 deuterated-benzene:carbon tetrachloride and enantiomeric excess determined by chiral shift n.m.r.","techniques employing a europium tris[3-(heptafluoropropylhydroxy-methylene) -(+)-camphorate] complex.","Table 2 illustrates the enantiomeric excess observed in the cyclopropane compound prepared in 2(a).","In each case it can be seen that there is a significant enantiomeric excess indicating that the reaction product has enhanced chirality.","TABLE 2 Catalyst Enantiomeric Enantiomeric Catalyst Number Concentration Ratio Excess 9a 10 57/43 14 9b 7.5 72/28 44 9c 7.5 33/67 34 9d 7.5 27/73 46 9e 7.5 64/36 28 9f 7.5 21/79 58 9g 7.5 80/20 60 9h 7.5 35/65 30 9i 10 73/27 46 9j 10 27/73 46 9k 10 29/71 42 9l 10 73/27 46 3 Comparison of Cyclopropane Preparation from Various 1,2-Dioxines Cyclopropane compounds according to the invention were prepared from various 1,2-dioxine starting materials utilising catalysts 9b, 9i and 9l as identified above and the degree of chirality in the cyclopropane products compared by measurement of the enantiomeric excess as described above.","The 3,6-disubstituted-3,6-dihydro-1,2-dioxines and the benzyl ester ylide were reacted in dichloromethane at 20-22° C. with 7.5-10 mol % catalyst.","The following symmetrical 1,2-dioxines were used: 1a 3,6-Diphenyl-3,6-dihydro-1,2-dioxine (X═Y═Ph) 1b 3,6-Dipropyl-3,6-dihydro-1,2-dioxine (X═Y═n-propyl) 1c 3,6-Dinapthyl-3,6-dihydro-1,2-dioxine (X═Y═1-napthyl) In each case the catalyst used results in a product having enhanced enantioselectivity for range or 1,2-dioxines, as illustrated in Table 3.A diverse range of aryl and alkyl substituents in the 1,2-dioxine would produce a similar effect.","TABLE 3 Catalyst Catalyst Enantiomeric Enantiomeric 1,2-Dioxine Number Conc.","Ratio Excess 1a 9b 7.5 72/28 44 9i 10 73/27 46 9l 10 73/27 46 1b 9b 10 67/33 34 9i 10 66/34 32 9l 10 65/35 30 1c 9b 10 72/28 44 4.Cyclopropane Preparation Using Chiral Salen Based Catalyst (8a-8e) Cobalt (II) Salens of the type depicted in 8a-8e were used as catalyst in the reaction between 3,6-diphenyl-3,6-dihydro-1,2-dioxine (1a) in the presence of a benzyl ester ylide conducted in dichloromethane or tetrahydrofuran (THF) at 20-22° C. with 2-5% mol % catalyst.","The enantiomeric excess of the resulting cyclopropane product was measured by the technique described hereinabove.","Results are tabulated in table 4 and indicate that the catalysts 8a to 8h induce enantioselectivity into the cyclopropane product for a range of symmetrical 1,2-dioxines.","It is also interesting to note that, in the case of catalyst 8h, the enantiomeric excess is increased when THF is used as a solvent compared with dichloromethane.","In the case of catalysts 8f, 8g and 8h it can be seen that these catalysts contain more than one chiral centre.","TABLE 4 Catalyst Enan- Catalyst 1,2- Concen- Enantiomeric tiomeric Number Solvent dioxine tration Ratio Excess 8a Dichloromethane 1a 2 35/65 30 8b Dichloromethane 1a 2 31/69 38 8c Dichloromethane 1a 2 31/69 38 8d Dichloromethane 1a 2 38/62 24 8e Dichloromethane 1a 2 36/64 28 8a Dichloromethane 1b 2 40/60 20 8b Dichloromethane 1b 2 35/65 30 8c Dichloromethane 1b 2 36/64 28 8d Dichloromethane 1b 2 41/59 18 8e Dichloromethane 1b 2 41/59 18 8b THF 1a 5 28/72 44 8c THF 1a 5 34/65 32 8d THF 1a 5 27/73 46 8e THF 1a 5 36/64 28 8f dichloromethane 1a 5 35/65 30 8g dichloromethane 1a 5 33/67 34 8h dichloromethane 1a 5 75/25 50 8h THF 1a 5 82/18 64 5.Effect of Catalyst Concentration on the Enantiomeric Excess (a) Effect of a Beta-Ketoiminato Catalyst In the reaction between 3,6-diphenyl-3,6-dihydro-1,2-dioxine with benzyl ester ylide at 20° in dichloromethane the effect of various concentrations of the catalyst 9b on the enantiomeric excess was measured.","20 mg 1,2-dioxine was employed for 1 mL of solvent.","Table 5a gives details of the measured enantiomeric ratio and enantiomeric excess.","Attention should be drawn to the fact that in the absence of the catalyst (catalyst mol %=0) the reaction product is a racemic mixture with no enantiomeric excess, thereby indicating the effect of the catalyst on the process.","The catalyst is most effective in the range 7.5 mol %.","TABLE 5a CATALYST (MOL %) ENANTIOMERIC Ratio ENANTIOMERIC EXCESS 0 50/50 0 2.5 58/42 16 5 69/31 38 7.5 73/27 46 10 71/29 42 15 70/30 40 20 69/31 38 35 70/30 40 50 60/40 20 (b) Effect of a Beta-Ketoiminato Catalyst in THF or Dichloromethane Solvents TABLE 5b CAT- ENANTIOMERIC ENANTIOMERIC Solvent ALYST Ratio EXCESS THF 9b 84/16 68 dichloromethane 9b 72/28 44 THF 9c 82/18 64 dichloromethane 9c 67/33 34 THF 9d 73/27 46 dichloromethane 9d 73/27 46 THF 9h 75/25 50 dichloromethane 9h 65/35 30 THF 9g 86/14 72 dichloromethane 9g 80/20 60 The reaction between 3,6-diphenyl-3,6-dihydro-1,2-dioxine and benzyl ester ylide at 20° C. in dichloromethane or THF was catalysed with the catalysts identified as 9b, 9c, 9d, 9h and 9g.","20 mg of 1,2-dioxine was employed for each 20 mL of solvent.","In each case the catalyst concentration was maintained at 7.5 mol %.","Results are shown in table 5b (c) Effect of Solvent on Catalyst Performance Reaction of Cobalt (II) catalyst 9b with 3,6-diphenyl-3,6-dihydro-1,2-dioxine (1a) in the presence of the benzyl ester ylide at 20° C. in a specific solvent.","20 mg 1,2-dioxine employed for 1 mL of solvent.","Employing 5.0 mol % catalyst.","Results are shown in table 5c TABLE 5c ENANTIOMERIC ENANTIOMERIC Solvent Ratio EXCESS acetonitrile 76/24 52 25% ethyl acetate/hexane 78/22 56 Ethyl acetate 77/23 54 THF 84/16 68 ether 67/33 34 toluene 67/33 34 acetone 76/24 52 Carbon tetrachloride 67/33 34 dichloromethane 68/32 36 It can be seen that that the cobalt catalyst 9b induces varying degrees of enantioselectivity in this reaction for a range of solvents under the conditions mentioned.","The optimum enantiomeric excess is induced when the THF was employed as solvent.","(d) Temperature Effects Reaction of Cobalt (II) catalyst with 3,6 diphenyl-3,6-dihydro-1,2-dioxine (1a) in the presence of the benzyl ester ylide at a specific temperate in a specific solvent.","20 mg 1,2-dioxine employed for 1 mL of solvent.","Employing 5.0 mol % catalyst.","Results are shown in table 5(d).","The comclusion may be drawn from the results shwn in Table 5d that changing the temperature has a dramatic effect on the degree of enantioselectivity in this reaction in certain solvents.","Lowering the temperature increase the observed enantiomeric excess.","TABLE 5d ENANTIO- Temperature MERIC ENANTIOMERIC Solvent (° C.) Ratio EXCESS dichloromethane 20 69/31 38 dichloromethane 0 74/26 48 dichloromethane −10 78/22 56 dichloromethane −15 81/19 62 dichloromethane −20 86/14 72 dichloromethane −40 80/20 60 THF 20 84/16 68 THF 0 87/13 74 THF −15 88/12 76 THF −40 86/14 72 5 Preparation of Optically Enriched Benzyl 2-[2-Benzoyl-3-Phenylcyclopropyl]acetate (a) Preparation Using Benzyl Ester Ylide (i) Procedure A solution of 3,6-diphenyl-3,6-dihydro-1,2-dioxine (0.5 g) in THF (20 mL) and a solution of catalyst 9b (120 mg) in THF (5 mL) were equilibrated at −15° C. for 30 minutes after which time the two solutions were combined and the temperature maintained at−15° C. The rearrangement to the γ-hydroxyenone was monitored by t.l.c.","and when complete the benzyl ester ylide (1.0 g) was added and the reaction mixture stirred overnight.","The solvent was then removed in vacuo and the residue purified by chromatography (silica) (3:17 ethyl acetate:hexanes).","The enantiomeric excess was determined to be 72% (an enantiomeric ratio of 86/14).","The isolated yield of the benzyl 2-[2-benzoyl-3-phenylcyclopropyl]acetate product vias determined to be 78%.","(ii) Recrystallization The enriched optical isomer of the benzyl 2-[2-benzoyl-3-phenylcyclopropyl]acetate prepared in step (a) can be further enriched by conventional processes used to isolate optical isomers.","Recrystallization of the product from hexane:dichloromethane (10:1) at ambient temperatures resulted in a Product having an enantiomeric ratio of 88/12.","(b) Preparation Using a Tert-Butyl Ester Ylide Reaction of Cobalt (II) catalyst 9b with 3,6diphenyl-3,6-dihydro-1,2-dioxine (1a) in the presence of the tert-butyl ester ylide (a bulky ylide) at a specific temperature in a specific solvent.","Experimental Procedure for the Above Reaction A solution of catalyst 9b (12 mg, 0.5 mol %) in dichloromethane (0.1 mL) was added to an equilibrated solution of 3,6-diphenyl-3,6-dihydro-1,2-dioxine 1a (50 mg) in dichloromethane (2.9 mL) at 20° C. After rearrangement of the dioxine was complete lithium bromide (19 mg) was added, and allowed to stir for 5 minutes, followed by t-butyl ester ylide and the reaction mixture stirred vigorously for 5 days.","The reaction mixture was filtered and the solvent was then removed in vacuo.","Purification by chromatography (3:17 ethyl acetate:hexanes) gave the pure cyclopropane (40.1 mg, 57%) with an introduced enantiomeric excess of 69/31, 38%.","(b) Preparation of Cyclopropanes in D2O Cyclopropanes in accordance with the invention were prepared using catalysts of the invention as shown in scheme 5.A solution of catalyst 9b (12 mg, 7.5 mol %) in dichloromethane (0.1 mL) was added to an equilibrated solution of 3,6-diphenyl-3,6-dihydro-1,2-dioxine (50 mg) in dichloromethane (2.9 mL) at 20° C. After rearrangement of the dioxine was complete deuterium oxide (1 mL) was added, and allowed to stir vigorously for 1 hour, followed by benzyl ester ylide and the reaction mixture stirred vigorously for 5 days.","The deuterium oxide was removed employing a pipette off and the reaction mixture dried (Na2SO4).","The solvent was removed in vacuo and the residue purified by chromatography (3:17 ethyl acetate: hexanes) to give the pure cyclopropane (49.9 mg, 67%) with an introduced enantiomeric excess of 75/25, 50%.","6.Comparison of Cyclopropane Preparation from Various Phosphorus Ylides and Precursors Typical Procedure for the Preparation of Cyclopropane 5 Derived from the Combination of Phosphonate (12), Symmetrical 1,2-dioxine 1a and Cobalt Catalyst 9b.","To a solution of phosphonate 12 (45 mg) in THF (230 μl) at−78° C. was added methyl lithium (175 μl, 1.4 M) over 5 minutes and the reaction mixture stirred at−78° C. for 30 minutes and then allowed to attain ambient temperature.","Separately the 1,2-dioxine 1a (60 mg) was combined with the cobalt catalyst 9b (12 mg) in THF (3 mL) and the dioxine allowed to rearrange.","This, mixture was then cooled to −78° C. and the ylide generated above added drop wise via a syringe and the mixture stirred at −78° C. for 30 minutes before being allowed to attain ambient temperature.","After 15 hours the solvent was removed in vacuo and the residue purified by column chromatography (3:17 ethyl acetate/hexanes) to afford the cyclopropane 5 in 79% yield.","The ee was measured by chiral shift NM as described herein above.","The conditions used for the preparation of the cyclopropanes from the use of ylides of type 10 and 11 above are as described for the benzyl ester ylide.","Table 6: Enantiomeric excess observed in cyclopropanes of types 5, 13 and 14 depicted above made using symmetrical 1,2-dioxine 1a and ylides of type 10-12 (depicted above) in dichloromethane or tetrahydrofuran (THF) at 20-22° C. with 7.5 mol % of catalyst 9b.","Note: the ylide of the phosphonate 12 is generated first with base, see procedure above.","Catalyst Enantiomeric Enantiomeric Dioxine Ylide Number Solvent Ratio Excess 1a 10 9b dichloromethane 71/29 42 1a 11 9b dichloromethane 69/31 38 1a 12 9b THF 83/17 66 The catalysts and processes of the invention therefore enable the synthesis of a range of isotopically labelled cyclopropane compounds, which are useful in a variety of situations.","7.Synthetic Schemes Using Optically Enriched Cyclopropanes Scheme 6 illustrates an example of how an enantiomerically enhanced cyclopropane produced in the process of the present invention can be used in the synthesis of optically enriched amide products.","(a) Preparation of Trans (±) 2-[2-Benzoyl-3-Phenylcyclopropyl]Acetic Acid Hydrolysis of cyclopropane 5 (ee ratio 86/14) prepared above afforded the acid (15).","The acid product was found to have a similar ee to the cyclopropane starting material, that is, the material is optically enhances.","(b) Preparation of Trans 2-[2-Benzoyl-3-Phenylcycloproyl]Acetyl Chloride To a solution of trans 2-[2-benzoyl-3-phenylcyclopropyl]acetic acid (15) (0.394 g) of ee cited above in dry dichloromethane 8 mL) and DMF (1 drop), at 0° C. under nitrogen, was added oxalyl chloride (0.291 g) in dichloromethane (4 mL) drop wise over 5 minutes.","The reaction mixture was then stirred at 0° C. for 2 hours and a further 2 hours at room temperature.","The volatiles were then removed to afford crude (16) which was used as is.","(c) Preparation of Optically Pure (4S)3-{2-[1S, 2R, 3R)-2-Benzoyl-3-Phenylcyclopropyl]Acetyl}-4-benzyl-1,3-oxazolan-2-one and (4S)3-{2-[(1R, 2S, 3S)-2-Benzoyl-3-Phenylcyclopropyl]Acetyl}-4-benzyl-1,3-oxazolan-2-one.","To a solution of (S)-4-benzyl-2-oxazolidinone (17) (0.252g) in THF (4 mL) with triphenylmethane (5 mg, indicator), at−78° C. under nitrogen, was added n-BuLi (1.9M in hexanes) until an orange colour persisted.","The reaction mixture was then stirred at−78° C. for 1 hour after which time the acid chloride in THF (3 mL) was added via a cannula.","The reaction was then stirred at −78° C. for 4 hours then poured into NB4Cl (sat.","10 mL) immediately, diluted with water (10 mL) and extracted with dichloromethane (2×30 mL).","The combined organic phases were dried (Na2SO4) and the solvent removed in vacuo.","The residue was purified by chromatography (1:3, ethyl acetate:hexanes) with separation of the diastereomers.","The two enantiomers were characterised as follows: First Enantiomer eluted (top spot) Rf 0.20 (1:5 ethyl acetate:hexanes).","1H NMR (600 MHz) δ 2.42-2.46 (m, 1H), 2.67 (dd, 1H, J=13.2, 9.6 Hz), 2.93 (dd, 1H, J=17.4, 7.2 Hz), 2.99 (dd, 1H, J=17.4, 7.8 Hz), 3.16 (dd, 1H, J=9.6, 4.8 Hz), 3.23 (dd, 2H, J=9.6, 4.8 Hz), 4.06 (t, 1H, J=8.4 Hz), 4.08 (dd, 1H, J=9.3 Hz), 4.51-4.55 (m, 1H), 7.15 (d, 2H, J=7.2 Hz), 7.22-7.26 (m, 2H), 7.28-7.34 (m, 6H), 7.52 (t, 2H, J=7.8 Hz), 7.61 (tt, 1H, J=7.2, 1.2 Hz), 8.08-8.10 (m, 2H).","13C NMR δ 27.16, 29.01, 33.60, 33.83, 37.81, 55.08, 66.18, 126.93, 127.36, 128.19, 128.43, 128.69, 128.79, 128.94, 129.35, 133.05, 135.11, 135.88, 137.72, 171.61.IR (dichloromethane solution) 1668, 1703, 1780 cm−1.MS m/z 440.2 (42%, [M+H]+), 422.2 (8%), 263.1 (100%), 221.1 (37% o).","C28H25NO4 requires C: 76.51, H: 5.74, found C: 76.63, H: 5.65.Accurate MS C28H25NO4 requires 439.17836, found 440.18753 ([M+F]+); [α]24=−39.24 (c=2.06, CH2Cl2)7 Second Enantiomer eluted (bottom spot) Rf 0.13 (1:5 ethyl acetate:hexanes).","1H NMR (600 MHz) δ 2.46 (tt, 1H, J=7.8, 6.6 Hz), 2.59 (dd, 1H, J=13.8, 9 Hz), 2.93 (dd, 1H, J=18, 6.6 Hz), 2.99 (ddI, 1H, J=17.4, 7.8 Hz), 3.05 (dd, 1H, J=13.8, 3.6 Hz), 3.21 (d, 2H, J=6H), 4.09 (dd, 1H, J=9, 3 Hz), 4.13 (t, 1H, J=7.8 Hz), 4.60-4.64 (m, 1H), 7.02 (dd, 2H, J=7.8, 1.8 Hz), 7.24-7.28 (m, 4H), 7.33 (d, 4H, J=4.2 Hz), 7.51 (t, 2H, J=7.8 Hz), 7.59 (tt, 1H, J=7.8, 1.8 Hz), 8.09-8.10 (m, 2H).","13C NMR (600M) δ 26.94, 28.81, 33.75, 34.31, 37.47, 54.75, 66.09, 127.02, 127.34, 128.16, 128.48, 128.67, 128.88, 128.92, 129.35, 133.04, 134.87, 135.88, 137.72, 153.30, 171.45, 198.39.IR (dichloromethane solution) 1668, 1703, 1782 cm−1.MS m/z 440.1 (45%, [M+H]+), 422.1 (10%), 263.1 (100%), 221.1 (34%).","C28H25NO4 requires C: 76.51, H: 5.74, found C: 76.49, H: 5.68.Accurate MS C28H25NO4 requires 439.17836, found 440.18712 ([M+H]+).","The invention has been described by way of example.","The examples are not, however, to be taken as limiting the scope of the invention in any way.","Modifications and variations of the invention such as would be apparent to a skilled addressee are deemed to be within the scope of the invention."]],"string":"[\n [\n \"Enantiomerically selective cyclopropanation\",\n \"A method of forming a cyclopropane having enhanced chirality said method comprising reacting together: a symmetrical 1,2-dioxine of the formula (1), wherein X and Y are the same and are groups in which a carbon atom is bonded to the dioxine backbone; and a phosphorus ylide or a phosphorus ylide precursor, in the presence of a cobalt catalyst containing a chiral ligand.\"\n ],\n [\n \"1.A method of forming a cyclopropane having enhanced chirality said method comprising reacting together: i) a symmetrical 1,2-dioxine of the formula (1), wherein X and Y are the same and are groups in which a carbon atom is bonded to the dioxine backbone; and ii) a phosphorus ylide or a phosphorus ylide precursor; in the presence of iii) a cobalt catalyst containing a chiral ligand.\",\n \"2.A method of forming a cyclopropane having enhanced chirality according to claim 1, wherein the chiral ligand in the catalyst has the structure: —(OR′C═R′CR′C═NR′CHCHR′N═CR′CR′═COR′)— around the cobalt atom and wherein each R′ moiety may be independently selected from a wide range of groups including, but not limited to H, alkyl, aryl, keto, ester and a range of other functionalities.\",\n \"3.A method of forming a cyclopropane having enhanced chirality according to claim 1, in which the cobalt catalyst has the structure: Wherein R1 and R2 may be hydrogen, alkyl, aryl; R3 may be hydrogen, alkyl, t-alkyl, alkyl or aryl, alkoxy, aryl alkoxy; R3, R4, R5 and R6 may be independently H, alky, aryl, keto, or ester.\",\n \"4.A method of forming a cyclopropane having enhanced chirality according to claim 3, in which R1 and R2 are each the same and are selected from hydrogen, (CH2)4, or benzyl.\",\n \"5.A method of forming a cyclopropane having enhanced chirality according to claim 3, in which R3 is hydrogen, t-butyl, or benzyl ethyl, bornoxy, menthoxy, ethoxy or methyl.\",\n \"6.A method of forming a cyclopropane having enhanced chirality according to claim 3, in which R4 and R5 may be the same or different and are selected from hydrogen, and t-butyl.\",\n \"7.A method of forming a cyclopropane having enhanced chirality according to claim 1 in which the phosphorus ylide or phosphorus ylide precursor may be taken to mean a stabilized phosphorus ylide or a reactive compound capable of forming a phosphorus ylide in situ.\",\n \"8.A method of forming a cyclopropane having enhanced chirality according to claim 1, in which the phosphorus ylide or phosphorus ylide precursor is a compound of the formula: i) R7R8R9P=CZCO2R10; wherein R7, R8 and R9 may be the same or different and may be alkyl, substituted alkyl, aryl, substituted aryl; R10 represents a non bulky group such ac C1-4 alkyl or substituted alky, or a bulky group such as 1-adamantyl; and Z represents hydrogen or methyl; or ii) R7R8R9P═CONR10R11; wherein R7, R8, R9 and R10 have the values identified in (i) above and wherein R11 represents a C1-12 alkoxy grouping; iii) R7R8R9P═CN; or iv) (R70)2P(O)CH2CO2 R10 wherein R7 and R10 have the values identified in (i) above.\",\n \"9.A method of forming a cyclopropane having enhanced chirality according to claim 1, in which the phosphorus ylide is a non-bulky stabilized ylide.\",\n \"10.A method of forming a cyclopropane having enhanced chirality according to claim 9, in which the non-bulky stabilized ylide is benzyl 2-(triphenyl-λ-5 phosphanylidene) acetate.\",\n \"11.A method of forming a cyclopropane having enhanced chirality according to claim 1, in which the phosphorus ylide is a bulky ylide.\",\n \"12.A method of forming a cyclopropane having enhanced chirality according to claim 1, in which the bulky ylide is t-butyl 2-(triphenyl-λ-phosphanylidene) acetate.\",\n \"13.A method of forming a cyclopropane having enhanced chirality according to claim 1, in which the phosphorus ylide or phosphorus ylide precursor is selected from N,N-methoxymethyl-2-(1,1,1-triphenyl-λ5-phosphanylidene)acetamide; 2-(1,1,1-triphenyl-λ5-phosphanylidene) acetonitrile or methyl 2-(dimethoxyphosphoryl) acetate.\",\n \"14.A method of forming a cyclopropane having enhanced chirality according to claim 1, in which X and Y are selected from H, alkyl or aryl groups.\",\n \"15.A method of forming a cyclopropane having enhanced chirality according to claim 1, wherein the reaction is conducted in a solvent selected from acetonitrile, ethyl acetate/hexane admixtures, ethyl acetate, tetrahydrofuran, ether, toluene, acetone, carbon tetrachloride, and dichloromethane or mixtures thereof.\",\n \"16.A method of forming a cyclopropane having enhanced chirality according to claim 1, in which the reaction is conducted with catalyst concentration of up to 50 mol % catalyst relative to the 1,2-dioxine, and more preferably a catalyst concentration of 1-15 mol %.\",\n \"17.A method of forming a cyclopropane having enhanced chirality according to claim 1, wherein the optically enriched cyclopropanes are subjected to recrystallization or column chromatography.\",\n \"18.A method of forming a cyclopropane having enhanced chirality according to claim 1, in which the enhanced chirality of the product, as determined by chiral shift n.m.r.\",\n \"techniques and expressed as an enantiomeric ratio may be as high as 90/10.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>Cyclopropanes containing natural and non-natural products are receiving considerable attention as synthetic targets since the incorporation of the rigidified cyclopropyl skeleton into bioactive analogues leads to conformationally constrained molecules.\",\n \"Such modifications often have significant effects on bioactivities with concomitant medical implications.\",\n \"Cyclopropanes are also generated as transient species in primary and secondary metabolisms in man, plants and microorganisms.\",\n \"Thus, the great importance of functionalised cyclopropanes in organic synthesis spurs a continuing search for efficient stereo controlled cyclopropanation methodologies.\",\n \"Of particular importance is the deficiency in methods for the construction of diversely functionalised cyclopropanes that contain greater than di-substitution.\",\n \"Chemicals containing the cyclopropyl moiety may be used inter alia: in the preparation of cyclopropyl amino acids; the preparation of cyclopropyl pharmaceuticals; the preparation of cyclopropyl pesticides; the preparation of new cyclopropyl products for furrier biological evaluation by chemistry and biochemistry laboratories as well as the preparation of isotopically labelled cyclopropanes as bioactive probes.\",\n \"Cyclopropanes may be synthesised by a number of techniques including (i) the direct carbene transfer (both stoichiometric and catalytic) from a diazo precursor to an olefin utilizing transition metals (Rh, Cu, Zn, and Pd) and (ii) Michael addition of nucleophiles (usually sulphur ylides) to α, β-unsaturated ketones and esters followed by intra-molecular cyclization.\",\n \"Such methods, employing as they do, potentially explosive material or requiring reaction mixtures that are highly sensitive to moisture have certain practical drawbacks.\",\n \"An alternative reaction scheme for the synthesis of cyclopropanes involves the reaction between 1,2-dioxines and stabilised phosphorus ylides, is illustrated in reaction scheme 1.It is suggested that the ylide acts as a mild base inducing the ring opening of the 1,2-dioxine (1) to their isomeric cis γ-hydroxy enones (3) followed by Michael addition of the y]ide to the enone and attachment of the electrophilic phosphorus pole of the ylide to the hydroxyl moiety affording the intermediate 1-2λ-oxaphospholanes (4).\",\n \"Cyclization of the resultant enolate produces th cyclopropanes (5) in excellent yield.\",\n \"The cyclopropane products emerging from this reaction scheme have in the past been prepared as racemic mixtures.\",\n \"The present invention is directed to a synthetic route for the preparation of optically enriched cyclopropane compounds.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>Therefore according to a first aspect of the invention there is provided a method of forming a cyclopropane having enhanced chirality said method comprising reacting together: (i) a symmetrical 1,2-dioxine of the formula (1), wherein X and Y are the same and are groups in which a carbon atom is bonded to the dioxine backbone; and (ii) a phosphorus ylide or a phosphorus ylide precursor; in the presence of (iii) a cobalt catalyst containing a chiral ligand.\",\n \"Preferably, the cobalt catalyst hils one of the structures shown below wherein R1 and R2 may be hydrogen, alkyl, aryl; R3 may be hydrogen, alkyl, t-alkyl, alkyl or aryl, alkoxy, aryl alkoxy; R3, R4, R5 and R6 may be independently H, alkyl, aryl, keto, or ester In accordance with a further aspect of the invention there is provided a chiral cyclopropanation catalyst of the formulae shown below in Table 1 based on Cobalt Salen compounds and on Cobalt Beta-Ketoiminato compounds.\"\n ],\n [\n \"FIELD OF THE INVENTION The present invention relates to a process for introducing an asymmetric cyclopropyl group into a molecule BACKGROUND OF THE INVENTION Cyclopropanes containing natural and non-natural products are receiving considerable attention as synthetic targets since the incorporation of the rigidified cyclopropyl skeleton into bioactive analogues leads to conformationally constrained molecules.\",\n \"Such modifications often have significant effects on bioactivities with concomitant medical implications.\",\n \"Cyclopropanes are also generated as transient species in primary and secondary metabolisms in man, plants and microorganisms.\",\n \"Thus, the great importance of functionalised cyclopropanes in organic synthesis spurs a continuing search for efficient stereo controlled cyclopropanation methodologies.\",\n \"Of particular importance is the deficiency in methods for the construction of diversely functionalised cyclopropanes that contain greater than di-substitution.\",\n \"Chemicals containing the cyclopropyl moiety may be used inter alia: in the preparation of cyclopropyl amino acids; the preparation of cyclopropyl pharmaceuticals; the preparation of cyclopropyl pesticides; the preparation of new cyclopropyl products for furrier biological evaluation by chemistry and biochemistry laboratories as well as the preparation of isotopically labelled cyclopropanes as bioactive probes.\",\n \"Cyclopropanes may be synthesised by a number of techniques including (i) the direct carbene transfer (both stoichiometric and catalytic) from a diazo precursor to an olefin utilizing transition metals (Rh, Cu, Zn, and Pd) and (ii) Michael addition of nucleophiles (usually sulphur ylides) to α, β-unsaturated ketones and esters followed by intra-molecular cyclization.\",\n \"Such methods, employing as they do, potentially explosive material or requiring reaction mixtures that are highly sensitive to moisture have certain practical drawbacks.\",\n \"An alternative reaction scheme for the synthesis of cyclopropanes involves the reaction between 1,2-dioxines and stabilised phosphorus ylides, is illustrated in reaction scheme 1.It is suggested that the ylide acts as a mild base inducing the ring opening of the 1,2-dioxine (1) to their isomeric cis γ-hydroxy enones (3) followed by Michael addition of the y]ide to the enone and attachment of the electrophilic phosphorus pole of the ylide to the hydroxyl moiety affording the intermediate 1-2λ-oxaphospholanes (4).\",\n \"Cyclization of the resultant enolate produces th cyclopropanes (5) in excellent yield.\",\n \"The cyclopropane products emerging from this reaction scheme have in the past been prepared as racemic mixtures.\",\n \"The present invention is directed to a synthetic route for the preparation of optically enriched cyclopropane compounds.\",\n \"SUMMARY OF THE INVENTION Therefore according to a first aspect of the invention there is provided a method of forming a cyclopropane having enhanced chirality said method comprising reacting together: (i) a symmetrical 1,2-dioxine of the formula (1), wherein X and Y are the same and are groups in which a carbon atom is bonded to the dioxine backbone; and (ii) a phosphorus ylide or a phosphorus ylide precursor; in the presence of (iii) a cobalt catalyst containing a chiral ligand.\",\n \"Preferably, the cobalt catalyst hils one of the structures shown below wherein R1 and R2 may be hydrogen, alkyl, aryl; R3 may be hydrogen, alkyl, t-alkyl, alkyl or aryl, alkoxy, aryl alkoxy; R3, R4, R5 and R6 may be independently H, alkyl, aryl, keto, or ester In accordance with a further aspect of the invention there is provided a chiral cyclopropanation catalyst of the formulae shown below in Table 1 based on Cobalt Salen compounds and on Cobalt Beta-Ketoiminato compounds.\",\n \"Chiral Cobalt Salen Based Catalysts 8a: R1,R2=—(CH2)4—, R3=t-Bu, R4=H, R5=t-Bu (R,R isomer) 8b: R1,R2=—(CH2)4—, R3=R4=R5=H (R,R isomer) 8c: R1,R2=—(CH2)4—, R3=t-Bu, R4=R5=H (R,R isomer) 8d: R1,R2=Ph, R3=R4=R5=H (R,R isomer) 8e: R1,R2=Ph, R3=t-Bu, R4=R5=H (R,R isomer) 8f: R1,R2=—(CH2)4—, R3=S-PhEtCH—,R4=R5=H (R,R isomer) 8g: R1,R2=Ph, R3=S-PhEtCH—, 14=R5=H (R,R isomer) 8h: R1,R2=Ph, R3=S-PhEtCH—, 14=R5H (S,S isomer) Beta-Ketoiminato Cobalt Based Catalysts 9a: R1=R2=H, R6=(−)-bornoxy 9b: R1=R2=Ph, R6=(−)-bornoxy (S,S isomer) 9c: R1=R2=Ph, R6=(−)-bornoxy (R,R isomer) 9d: R1=R2=—(CH2)4—, R6=(−)-bornoxy(R,R isomer) 9e: R1=R2=Ph, R6=(−)-menthoxy (S,S isomer) 9f: R1=R2=Ph, R6=(−)-menthoxy (R,R isomer) 9g: R1=R2=Ph, R6=(+)-menthoxy (S,S isomer) 9h: R1=R2=Ph, R6=(+)-menthoxy (R,R isomer) 9i: R1=R2=Ph, R6=ethoxy (S,S isomer) 9j: R1=R2=Ph, ethoxy (R,R isomer) 9k: R1=R2=—(CH2)4—, R6=ethoxy (R,R isomer) 9l: R1=R2=Ph, R6=methyl (S,S isomer) Table 1 Catalysts according to the invention in the forms 8a to 8h may be prepared by reaction of Cobalt acetoacetate with the relevant SALEN ligand.\",\n \"SALEN ligands may be prepared by reaction of (1S, 2S) or (1R, 2R)-1,2-diphenylethylenediamine and the corresponding salicyaldehydes in ethanol under nitrogen atmosphere.\",\n \"The catalysts 9a-9l may be prepared from reaction of (S,S) or (R,R)-1,2-diphenyl-ethylenediamine and the relevant β-diketone derivative, followed by subsequent reaction with Co acetate.\",\n \"Corresponding ligands can be made from the (1S, 2S) and (1R, 2R) cyclohexyl-1,2-diamine.\",\n \"It can be seen that each of the SALEN and the beta-ketoiminato based catalysts has a common structure around the cobalt atom, namely, —(OR′C═R′CR′C═NR′CHCHR′N═CR′CR′═COR′)—.\",\n \"The catalysts each include at least one chiral grouping.\",\n \"It is possible for more than one chiral grouping to exist in the catalyst and the values of R′ are wide.\",\n \"Each of the R′ groups may independently be H, alkyl, aryl, keto, ester and a range of other functionalities.\",\n \"In the context of the present invention a phosphorus ylide or a phosphorus ylide precursor may be taken to mean a stabilized phosphorus ylide or a reactive compound capable of forming a phosphorus ylide in site.\",\n \"In a still further aspect of the present invention there is provided a method of forming a cyclopropane having enhanced chirality as described above in which the phosphorus ylide or phosphorus ylide precursor is a compound of the formula: (i) R7R8R9P═CZCO2R10; wherein R7, R8 and R9 may be the same or different and may be alkyl, substituted alkyl, aryl, substituted aryl; R10 represents a non bulky group such as C1-4 alkyl or substituted alkyl, or a bulky group such as 1-adamantyl; and Z represents hydrogen or methyl; or (ii) R7R8R9P═CONR10R11; wherein R7, R8, R9 and R10 have the values identified in (i) above and wherein R11 represents a C1-12 alkoxy grouping; or (iii) R7R8R9P═CN; or (iv) (R7O)2P(O)CH2CO2 R10 wherein R7 and R10 have the values identified in (i) above.\",\n \"Phosphorus ylides or a phosphorus ylide precursors of type (i) are exemplified by compound (2); type (ii) by compound (10); type (iii) by compound (11); and type (iv) by compound (12).\",\n \"Preferably, the phosphorus ester ylide is a non-bulky stabilized ylide such as a benzyl 2-(triphenyl-λ-phosphanylidene) acetate (hereinafter referred to as a benzyl ester ylide), or, alternatively, a bulky ylide such as t-butyl 2-(triphenyl-λ-phosphanylidene) acetate (hereinafter referred to as a t-butyl ester ylide).\",\n \"Alternative phosphorus ylides or ylide precursors include N,N-methoxymethyl-2-(1,1-triphenyl-λ5-phosphanylidene)acetamide; 2-(1,1,1-triphenyl-λ5-phosphanylidene) acetonitrile or methyl 2-(dimethoxyphosphoryl)acetate.\",\n \"Examples of the reaction between the phosphorus ylides and phosphorus ylide precursors with 1,2-dioxines and a chiral cobalt catalyst and the resulting chiral cyclopropanes are shown below: Preferably, the cyclopropanes of the invention are prepared in a solvent such as acetonitrile, ethyl acetate/hexane admixtures, ethyl acetate, tetrahydrofuran, ether, toluene, acetone, carbon tetrachloride, and dichloromethane or mixtures thereof.\",\n \"In a particularly preferred form of the invention the cyclopropanes of the invention are prepared in tetrahydrofuran.\",\n \"It will be appreciated by those skilled in the art that a range of solvents would in fact be suitable for wise in conducting the reaction of the invention.\",\n \"In any specific case an optimum solvent can be identified by trial and experiment using the above solvents and others.\",\n \"However, it should also be noted that alcohol is an unsuitable solvent for the reaction.\",\n \"The enantiomerically enhanced cyclopropanes of the invention are preferably prepared with catalyst concentration of up to 50 mol % catalyst relative to the 1,2-dioxine, and more preferably still a catalyst concentration of 1-15 mol %.\",\n \"The optically enriched cyclopropanes prepared in accordance with the invention may be further enriched by conventional techniques.\",\n \"For example, by recrystallization or conversion to diastereoisomers which can be separated by conventional techniques, e.g.\",\n \"recrystallization or column chromatography.\",\n \"DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will now be described by way of a number of non limiting examples.\",\n \"It should be noted that the invention presents a synthetic route for optically enhanced cyclopropanes and that the range of structures capable of synthesis according to the process of the invention is not to be take as being limited to those structures described herein.\",\n \"An extremely wide range of cyclopropane structures may be manufactured according to the process of the invention.\",\n \"1.Catalyst Preparation Beta Ketoiminato Catalyst Preparation—The preparation of a chiral cobalt catalyst (9b) (a) Preparation of (−)-Bornyl Acetoacetate To a solution of ethyl acetoacetate (4.5 g, 0.0346 mol) in n-heptane (70 mL) was added (−)-borneol (5.34 g, 0.0346 mol) and sodium hydride (50 mg) and the reaction mixture refluxed under Deans Stark conditions for 2 days.\",\n \"The solvent was then removed in vacuo and the residue purified by chromatography on silica (1:9 ethyl acetate:hexanes).\",\n \"(6.16 g, 70%) Rf 0.32 (1:9 ethyl acetate:hexanes) 1H NMR δ 0.85 (s, 3H), 0.88 (s, 3H), 0.91 (s, 3H), 0.94-1.43 (m, 4H) 1.64-2.00 (m, 3H) 2.85 (s, 3H), 3.47 (s, 2H), 4.95 (ddd, 1H, J=15, 5.1, 3.3 Hz).\",\n \"13C NMR δ 13.35, 18.71, 19.57, 26.96, 27.89, 29.99, 36.51, 44.76, 47.77, 48.76, 50.29, 81.07, 90.10, 167.30.MS m/z 238 (42%, M+), 154 (11%), 137 (100%), 121 (10%), 95 (23%).\",\n \"(b)(−) Bornyl 2-Formyl-3-Oxobutanoate To the (−)-bornyl acetoacetate (2.0 g, 8.4×10−3 moles) was added N,N-dimethylformamide dimethyl acetal (2 g, 1.68×10−2 moles), the mixture stirred at room temperature for 2 hours, then cooled to 0° C. and methanolic sodium hydroxide (1N, 14 mL 1:l methanol:water) added.\",\n \"The reaction mixture was stirred for a further 2 hours then cooled to 0° C. and hydrochloric acid (1N) added till pH 3-4.The mixture was extracted with diethyl ether, dried (Na2SO4) and the solvent removed in vacuo.\",\n \"Rf 0.6 (1:9 ethyl acetate:hexanes) Due to the instability of the aldehyde it was used directly in the next reaction to form the ligand.\",\n \"1H NMR δ 0.875 (s, 3H), 0.90 (s, 3H), 0.94 (s, 3H), 1.00-1.48 (m, 1H), 1.12-1.48 (m, 4H), 2.32-2.54 (m, 1H), 2.57 (s, 3H), 4.96-5.05 (m, 1H), 9.25 (d, 1H, J-9 Hz).\",\n \"The crude (−)-bornyl 2-formyl-3-oxobutanoate (0.25 g, 9.49×10−4 mol) was combined in ethanol (3 mL) with (1S, 2S)-1,2-diphenylethylenediamine (0.10 g, 4.75×10−4 mol) and allowed to react for 2 days, the solvent was then removed in vacuo and the residue purified by chromatography (1:9 acetone:dichloromethane) (290 mg, 86%).\",\n \"Rf 0.64 (1:9 acetone:dichloromethane).\",\n \"1H NMR δ0.79 (s, 6H), 0.86 (s, 6H), 0.90 (s, 6H), 0.96-1.32 (m, 4H), 1.60-1.82 (m, 6H), 2.28-2.49 (m, 2H), 2.51 (s, 6H), 4.67 (d, 2H, J=7.8 Hz), 4.89 (dm, 2H, J=8.2 Hz), 7.10-7.15 (m, 4H), 7.27-7.33 (m, 6H), 7.79 (d, 2H, J=12.8 Hz), 11.93-12.02 (m, 2H).\",\n \"13C NMR δ 13.63, 18.84, 19.70, 27.64, 28.03, 31.02, 36.92, 44.84, 47.74, 48.74, 69.53, 79.48, 102.05, 135.68, 158.96, 167.09, 199.71.MS m/z 710 (M+, 4%), 555 (22%), 147 (19%), 354 (14%), 200 (31%), 137 (33%), 95 (100%).\",\n \"(c) Preparation of Catalyst 9b) A mixture of ligand (144 mg, 2.03×10−4 mol) was combined with cobalt acetate tetrahydrate (50.6 mg, 2.03×10−4) in deaerated ethanol (4 mL) and heated under reflux for 4 hours.\",\n \"The solvent was then removed in vacuo till dryness of the complex resulted.\",\n \"The complex was used without further purification.\",\n \"Each of the catalysts 9a-9l may be prepared using similar methodology and by substitution of the appropriate bornoxy, ethoxy, methoxy, ethoxy or methyl starting materials.\",\n \"2.Cyclopropane Preparation Using Chiral Beta Ketoiminato Based Catalyst (9a-9l) (a) Preparation of Benzyl 2-(2-Benzoyl-3Phenylcyclopropyl) Acetate 3,6-Diphenyl-3,6dihydro-1,2 dioxine (1a) was reacted with the benzyl 2-(triphenyl-λ5-phosphanylidene)acetate (referred to as benzyl ester ylide) of in the presence of Cobalt (II) catalysts of type 9a-9l with as depicted in scheme 2 to produce the cyclopropane product.\",\n \"The cobalt catalyst 9b (4.8 mg, 6.3×10−6 mol, 5 mol %) was dissolved in dichloromethane (1 mL) and allowed to equilibrate at the temperature of the reaction.\",\n \"3,6-Diphenyl-3,6-dihydro-1,2-dioxine (20 mg, 8.4×10−5 mol) was added and the reaction left until such time as complete rearrangement of the dioxine had occurred (t.l.c.).\",\n \"Benzyl ester ylide (40 mg, 9.8×10−5 mol) was then added and the reaction mixture left for 10 hours.\",\n \"The solvent was then removed and the residue purified by column chromatography (SiO2, 3:17 ethyl acetate:hexanes).\",\n \"It is to be noted that the scope of the, invention is not limited to the specific 1,2-dioxines listed in scheme 3.A much greater range of substrates may be used to form a wide variety of substituted cyclopropane products.\",\n \"It should however be noted that the 1,2-dioxine must be a symmetrical dioxine.\",\n \"(b) Determination of Enantiomeric Excess Utilising the cyclopropane synthesised in 2(a) above, (5 mg) was dissolved in 1:4 deuterated-benzene:carbon tetrachloride and enantiomeric excess determined by chiral shift n.m.r.\",\n \"techniques employing a europium tris[3-(heptafluoropropylhydroxy-methylene) -(+)-camphorate] complex.\",\n \"Table 2 illustrates the enantiomeric excess observed in the cyclopropane compound prepared in 2(a).\",\n \"In each case it can be seen that there is a significant enantiomeric excess indicating that the reaction product has enhanced chirality.\",\n \"TABLE 2 Catalyst Enantiomeric Enantiomeric Catalyst Number Concentration Ratio Excess 9a 10 57/43 14 9b 7.5 72/28 44 9c 7.5 33/67 34 9d 7.5 27/73 46 9e 7.5 64/36 28 9f 7.5 21/79 58 9g 7.5 80/20 60 9h 7.5 35/65 30 9i 10 73/27 46 9j 10 27/73 46 9k 10 29/71 42 9l 10 73/27 46 3 Comparison of Cyclopropane Preparation from Various 1,2-Dioxines Cyclopropane compounds according to the invention were prepared from various 1,2-dioxine starting materials utilising catalysts 9b, 9i and 9l as identified above and the degree of chirality in the cyclopropane products compared by measurement of the enantiomeric excess as described above.\",\n \"The 3,6-disubstituted-3,6-dihydro-1,2-dioxines and the benzyl ester ylide were reacted in dichloromethane at 20-22° C. with 7.5-10 mol % catalyst.\",\n \"The following symmetrical 1,2-dioxines were used: 1a 3,6-Diphenyl-3,6-dihydro-1,2-dioxine (X═Y═Ph) 1b 3,6-Dipropyl-3,6-dihydro-1,2-dioxine (X═Y═n-propyl) 1c 3,6-Dinapthyl-3,6-dihydro-1,2-dioxine (X═Y═1-napthyl) In each case the catalyst used results in a product having enhanced enantioselectivity for range or 1,2-dioxines, as illustrated in Table 3.A diverse range of aryl and alkyl substituents in the 1,2-dioxine would produce a similar effect.\",\n \"TABLE 3 Catalyst Catalyst Enantiomeric Enantiomeric 1,2-Dioxine Number Conc.\",\n \"Ratio Excess 1a 9b 7.5 72/28 44 9i 10 73/27 46 9l 10 73/27 46 1b 9b 10 67/33 34 9i 10 66/34 32 9l 10 65/35 30 1c 9b 10 72/28 44 4.Cyclopropane Preparation Using Chiral Salen Based Catalyst (8a-8e) Cobalt (II) Salens of the type depicted in 8a-8e were used as catalyst in the reaction between 3,6-diphenyl-3,6-dihydro-1,2-dioxine (1a) in the presence of a benzyl ester ylide conducted in dichloromethane or tetrahydrofuran (THF) at 20-22° C. with 2-5% mol % catalyst.\",\n \"The enantiomeric excess of the resulting cyclopropane product was measured by the technique described hereinabove.\",\n \"Results are tabulated in table 4 and indicate that the catalysts 8a to 8h induce enantioselectivity into the cyclopropane product for a range of symmetrical 1,2-dioxines.\",\n \"It is also interesting to note that, in the case of catalyst 8h, the enantiomeric excess is increased when THF is used as a solvent compared with dichloromethane.\",\n \"In the case of catalysts 8f, 8g and 8h it can be seen that these catalysts contain more than one chiral centre.\",\n \"TABLE 4 Catalyst Enan- Catalyst 1,2- Concen- Enantiomeric tiomeric Number Solvent dioxine tration Ratio Excess 8a Dichloromethane 1a 2 35/65 30 8b Dichloromethane 1a 2 31/69 38 8c Dichloromethane 1a 2 31/69 38 8d Dichloromethane 1a 2 38/62 24 8e Dichloromethane 1a 2 36/64 28 8a Dichloromethane 1b 2 40/60 20 8b Dichloromethane 1b 2 35/65 30 8c Dichloromethane 1b 2 36/64 28 8d Dichloromethane 1b 2 41/59 18 8e Dichloromethane 1b 2 41/59 18 8b THF 1a 5 28/72 44 8c THF 1a 5 34/65 32 8d THF 1a 5 27/73 46 8e THF 1a 5 36/64 28 8f dichloromethane 1a 5 35/65 30 8g dichloromethane 1a 5 33/67 34 8h dichloromethane 1a 5 75/25 50 8h THF 1a 5 82/18 64 5.Effect of Catalyst Concentration on the Enantiomeric Excess (a) Effect of a Beta-Ketoiminato Catalyst In the reaction between 3,6-diphenyl-3,6-dihydro-1,2-dioxine with benzyl ester ylide at 20° in dichloromethane the effect of various concentrations of the catalyst 9b on the enantiomeric excess was measured.\",\n \"20 mg 1,2-dioxine was employed for 1 mL of solvent.\",\n \"Table 5a gives details of the measured enantiomeric ratio and enantiomeric excess.\",\n \"Attention should be drawn to the fact that in the absence of the catalyst (catalyst mol %=0) the reaction product is a racemic mixture with no enantiomeric excess, thereby indicating the effect of the catalyst on the process.\",\n \"The catalyst is most effective in the range 7.5 mol %.\",\n \"TABLE 5a CATALYST (MOL %) ENANTIOMERIC Ratio ENANTIOMERIC EXCESS 0 50/50 0 2.5 58/42 16 5 69/31 38 7.5 73/27 46 10 71/29 42 15 70/30 40 20 69/31 38 35 70/30 40 50 60/40 20 (b) Effect of a Beta-Ketoiminato Catalyst in THF or Dichloromethane Solvents TABLE 5b CAT- ENANTIOMERIC ENANTIOMERIC Solvent ALYST Ratio EXCESS THF 9b 84/16 68 dichloromethane 9b 72/28 44 THF 9c 82/18 64 dichloromethane 9c 67/33 34 THF 9d 73/27 46 dichloromethane 9d 73/27 46 THF 9h 75/25 50 dichloromethane 9h 65/35 30 THF 9g 86/14 72 dichloromethane 9g 80/20 60 The reaction between 3,6-diphenyl-3,6-dihydro-1,2-dioxine and benzyl ester ylide at 20° C. in dichloromethane or THF was catalysed with the catalysts identified as 9b, 9c, 9d, 9h and 9g.\",\n \"20 mg of 1,2-dioxine was employed for each 20 mL of solvent.\",\n \"In each case the catalyst concentration was maintained at 7.5 mol %.\",\n \"Results are shown in table 5b (c) Effect of Solvent on Catalyst Performance Reaction of Cobalt (II) catalyst 9b with 3,6-diphenyl-3,6-dihydro-1,2-dioxine (1a) in the presence of the benzyl ester ylide at 20° C. in a specific solvent.\",\n \"20 mg 1,2-dioxine employed for 1 mL of solvent.\",\n \"Employing 5.0 mol % catalyst.\",\n \"Results are shown in table 5c TABLE 5c ENANTIOMERIC ENANTIOMERIC Solvent Ratio EXCESS acetonitrile 76/24 52 25% ethyl acetate/hexane 78/22 56 Ethyl acetate 77/23 54 THF 84/16 68 ether 67/33 34 toluene 67/33 34 acetone 76/24 52 Carbon tetrachloride 67/33 34 dichloromethane 68/32 36 It can be seen that that the cobalt catalyst 9b induces varying degrees of enantioselectivity in this reaction for a range of solvents under the conditions mentioned.\",\n \"The optimum enantiomeric excess is induced when the THF was employed as solvent.\",\n \"(d) Temperature Effects Reaction of Cobalt (II) catalyst with 3,6 diphenyl-3,6-dihydro-1,2-dioxine (1a) in the presence of the benzyl ester ylide at a specific temperate in a specific solvent.\",\n \"20 mg 1,2-dioxine employed for 1 mL of solvent.\",\n \"Employing 5.0 mol % catalyst.\",\n \"Results are shown in table 5(d).\",\n \"The comclusion may be drawn from the results shwn in Table 5d that changing the temperature has a dramatic effect on the degree of enantioselectivity in this reaction in certain solvents.\",\n \"Lowering the temperature increase the observed enantiomeric excess.\",\n \"TABLE 5d ENANTIO- Temperature MERIC ENANTIOMERIC Solvent (° C.) Ratio EXCESS dichloromethane 20 69/31 38 dichloromethane 0 74/26 48 dichloromethane −10 78/22 56 dichloromethane −15 81/19 62 dichloromethane −20 86/14 72 dichloromethane −40 80/20 60 THF 20 84/16 68 THF 0 87/13 74 THF −15 88/12 76 THF −40 86/14 72 5 Preparation of Optically Enriched Benzyl 2-[2-Benzoyl-3-Phenylcyclopropyl]acetate (a) Preparation Using Benzyl Ester Ylide (i) Procedure A solution of 3,6-diphenyl-3,6-dihydro-1,2-dioxine (0.5 g) in THF (20 mL) and a solution of catalyst 9b (120 mg) in THF (5 mL) were equilibrated at −15° C. for 30 minutes after which time the two solutions were combined and the temperature maintained at−15° C. The rearrangement to the γ-hydroxyenone was monitored by t.l.c.\",\n \"and when complete the benzyl ester ylide (1.0 g) was added and the reaction mixture stirred overnight.\",\n \"The solvent was then removed in vacuo and the residue purified by chromatography (silica) (3:17 ethyl acetate:hexanes).\",\n \"The enantiomeric excess was determined to be 72% (an enantiomeric ratio of 86/14).\",\n \"The isolated yield of the benzyl 2-[2-benzoyl-3-phenylcyclopropyl]acetate product vias determined to be 78%.\",\n \"(ii) Recrystallization The enriched optical isomer of the benzyl 2-[2-benzoyl-3-phenylcyclopropyl]acetate prepared in step (a) can be further enriched by conventional processes used to isolate optical isomers.\",\n \"Recrystallization of the product from hexane:dichloromethane (10:1) at ambient temperatures resulted in a Product having an enantiomeric ratio of 88/12.\",\n \"(b) Preparation Using a Tert-Butyl Ester Ylide Reaction of Cobalt (II) catalyst 9b with 3,6diphenyl-3,6-dihydro-1,2-dioxine (1a) in the presence of the tert-butyl ester ylide (a bulky ylide) at a specific temperature in a specific solvent.\",\n \"Experimental Procedure for the Above Reaction A solution of catalyst 9b (12 mg, 0.5 mol %) in dichloromethane (0.1 mL) was added to an equilibrated solution of 3,6-diphenyl-3,6-dihydro-1,2-dioxine 1a (50 mg) in dichloromethane (2.9 mL) at 20° C. After rearrangement of the dioxine was complete lithium bromide (19 mg) was added, and allowed to stir for 5 minutes, followed by t-butyl ester ylide and the reaction mixture stirred vigorously for 5 days.\",\n \"The reaction mixture was filtered and the solvent was then removed in vacuo.\",\n \"Purification by chromatography (3:17 ethyl acetate:hexanes) gave the pure cyclopropane (40.1 mg, 57%) with an introduced enantiomeric excess of 69/31, 38%.\",\n \"(b) Preparation of Cyclopropanes in D2O Cyclopropanes in accordance with the invention were prepared using catalysts of the invention as shown in scheme 5.A solution of catalyst 9b (12 mg, 7.5 mol %) in dichloromethane (0.1 mL) was added to an equilibrated solution of 3,6-diphenyl-3,6-dihydro-1,2-dioxine (50 mg) in dichloromethane (2.9 mL) at 20° C. After rearrangement of the dioxine was complete deuterium oxide (1 mL) was added, and allowed to stir vigorously for 1 hour, followed by benzyl ester ylide and the reaction mixture stirred vigorously for 5 days.\",\n \"The deuterium oxide was removed employing a pipette off and the reaction mixture dried (Na2SO4).\",\n \"The solvent was removed in vacuo and the residue purified by chromatography (3:17 ethyl acetate: hexanes) to give the pure cyclopropane (49.9 mg, 67%) with an introduced enantiomeric excess of 75/25, 50%.\",\n \"6.Comparison of Cyclopropane Preparation from Various Phosphorus Ylides and Precursors Typical Procedure for the Preparation of Cyclopropane 5 Derived from the Combination of Phosphonate (12), Symmetrical 1,2-dioxine 1a and Cobalt Catalyst 9b.\",\n \"To a solution of phosphonate 12 (45 mg) in THF (230 μl) at−78° C. was added methyl lithium (175 μl, 1.4 M) over 5 minutes and the reaction mixture stirred at−78° C. for 30 minutes and then allowed to attain ambient temperature.\",\n \"Separately the 1,2-dioxine 1a (60 mg) was combined with the cobalt catalyst 9b (12 mg) in THF (3 mL) and the dioxine allowed to rearrange.\",\n \"This, mixture was then cooled to −78° C. and the ylide generated above added drop wise via a syringe and the mixture stirred at −78° C. for 30 minutes before being allowed to attain ambient temperature.\",\n \"After 15 hours the solvent was removed in vacuo and the residue purified by column chromatography (3:17 ethyl acetate/hexanes) to afford the cyclopropane 5 in 79% yield.\",\n \"The ee was measured by chiral shift NM as described herein above.\",\n \"The conditions used for the preparation of the cyclopropanes from the use of ylides of type 10 and 11 above are as described for the benzyl ester ylide.\",\n \"Table 6: Enantiomeric excess observed in cyclopropanes of types 5, 13 and 14 depicted above made using symmetrical 1,2-dioxine 1a and ylides of type 10-12 (depicted above) in dichloromethane or tetrahydrofuran (THF) at 20-22° C. with 7.5 mol % of catalyst 9b.\",\n \"Note: the ylide of the phosphonate 12 is generated first with base, see procedure above.\",\n \"Catalyst Enantiomeric Enantiomeric Dioxine Ylide Number Solvent Ratio Excess 1a 10 9b dichloromethane 71/29 42 1a 11 9b dichloromethane 69/31 38 1a 12 9b THF 83/17 66 The catalysts and processes of the invention therefore enable the synthesis of a range of isotopically labelled cyclopropane compounds, which are useful in a variety of situations.\",\n \"7.Synthetic Schemes Using Optically Enriched Cyclopropanes Scheme 6 illustrates an example of how an enantiomerically enhanced cyclopropane produced in the process of the present invention can be used in the synthesis of optically enriched amide products.\",\n \"(a) Preparation of Trans (±) 2-[2-Benzoyl-3-Phenylcyclopropyl]Acetic Acid Hydrolysis of cyclopropane 5 (ee ratio 86/14) prepared above afforded the acid (15).\",\n \"The acid product was found to have a similar ee to the cyclopropane starting material, that is, the material is optically enhances.\",\n \"(b) Preparation of Trans 2-[2-Benzoyl-3-Phenylcycloproyl]Acetyl Chloride To a solution of trans 2-[2-benzoyl-3-phenylcyclopropyl]acetic acid (15) (0.394 g) of ee cited above in dry dichloromethane 8 mL) and DMF (1 drop), at 0° C. under nitrogen, was added oxalyl chloride (0.291 g) in dichloromethane (4 mL) drop wise over 5 minutes.\",\n \"The reaction mixture was then stirred at 0° C. for 2 hours and a further 2 hours at room temperature.\",\n \"The volatiles were then removed to afford crude (16) which was used as is.\",\n \"(c) Preparation of Optically Pure (4S)3-{2-[1S, 2R, 3R)-2-Benzoyl-3-Phenylcyclopropyl]Acetyl}-4-benzyl-1,3-oxazolan-2-one and (4S)3-{2-[(1R, 2S, 3S)-2-Benzoyl-3-Phenylcyclopropyl]Acetyl}-4-benzyl-1,3-oxazolan-2-one.\",\n \"To a solution of (S)-4-benzyl-2-oxazolidinone (17) (0.252g) in THF (4 mL) with triphenylmethane (5 mg, indicator), at−78° C. under nitrogen, was added n-BuLi (1.9M in hexanes) until an orange colour persisted.\",\n \"The reaction mixture was then stirred at−78° C. for 1 hour after which time the acid chloride in THF (3 mL) was added via a cannula.\",\n \"The reaction was then stirred at −78° C. for 4 hours then poured into NB4Cl (sat.\",\n \"10 mL) immediately, diluted with water (10 mL) and extracted with dichloromethane (2×30 mL).\",\n \"The combined organic phases were dried (Na2SO4) and the solvent removed in vacuo.\",\n \"The residue was purified by chromatography (1:3, ethyl acetate:hexanes) with separation of the diastereomers.\",\n \"The two enantiomers were characterised as follows: First Enantiomer eluted (top spot) Rf 0.20 (1:5 ethyl acetate:hexanes).\",\n \"1H NMR (600 MHz) δ 2.42-2.46 (m, 1H), 2.67 (dd, 1H, J=13.2, 9.6 Hz), 2.93 (dd, 1H, J=17.4, 7.2 Hz), 2.99 (dd, 1H, J=17.4, 7.8 Hz), 3.16 (dd, 1H, J=9.6, 4.8 Hz), 3.23 (dd, 2H, J=9.6, 4.8 Hz), 4.06 (t, 1H, J=8.4 Hz), 4.08 (dd, 1H, J=9.3 Hz), 4.51-4.55 (m, 1H), 7.15 (d, 2H, J=7.2 Hz), 7.22-7.26 (m, 2H), 7.28-7.34 (m, 6H), 7.52 (t, 2H, J=7.8 Hz), 7.61 (tt, 1H, J=7.2, 1.2 Hz), 8.08-8.10 (m, 2H).\",\n \"13C NMR δ 27.16, 29.01, 33.60, 33.83, 37.81, 55.08, 66.18, 126.93, 127.36, 128.19, 128.43, 128.69, 128.79, 128.94, 129.35, 133.05, 135.11, 135.88, 137.72, 171.61.IR (dichloromethane solution) 1668, 1703, 1780 cm−1.MS m/z 440.2 (42%, [M+H]+), 422.2 (8%), 263.1 (100%), 221.1 (37% o).\",\n \"C28H25NO4 requires C: 76.51, H: 5.74, found C: 76.63, H: 5.65.Accurate MS C28H25NO4 requires 439.17836, found 440.18753 ([M+F]+); [α]24=−39.24 (c=2.06, CH2Cl2)7 Second Enantiomer eluted (bottom spot) Rf 0.13 (1:5 ethyl acetate:hexanes).\",\n \"1H NMR (600 MHz) δ 2.46 (tt, 1H, J=7.8, 6.6 Hz), 2.59 (dd, 1H, J=13.8, 9 Hz), 2.93 (dd, 1H, J=18, 6.6 Hz), 2.99 (ddI, 1H, J=17.4, 7.8 Hz), 3.05 (dd, 1H, J=13.8, 3.6 Hz), 3.21 (d, 2H, J=6H), 4.09 (dd, 1H, J=9, 3 Hz), 4.13 (t, 1H, J=7.8 Hz), 4.60-4.64 (m, 1H), 7.02 (dd, 2H, J=7.8, 1.8 Hz), 7.24-7.28 (m, 4H), 7.33 (d, 4H, J=4.2 Hz), 7.51 (t, 2H, J=7.8 Hz), 7.59 (tt, 1H, J=7.8, 1.8 Hz), 8.09-8.10 (m, 2H).\",\n \"13C NMR (600M) δ 26.94, 28.81, 33.75, 34.31, 37.47, 54.75, 66.09, 127.02, 127.34, 128.16, 128.48, 128.67, 128.88, 128.92, 129.35, 133.04, 134.87, 135.88, 137.72, 153.30, 171.45, 198.39.IR (dichloromethane solution) 1668, 1703, 1782 cm−1.MS m/z 440.1 (45%, [M+H]+), 422.1 (10%), 263.1 (100%), 221.1 (34%).\",\n \"C28H25NO4 requires C: 76.51, H: 5.74, found C: 76.49, H: 5.68.Accurate MS C28H25NO4 requires 439.17836, found 440.18712 ([M+H]+).\",\n \"The invention has been described by way of example.\",\n \"The examples are not, however, to be taken as limiting the scope of the invention in any way.\",\n \"Modifications and variations of the invention such as would be apparent to a skilled addressee are deemed to be within the scope of the invention.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10474149"}}},{"rowIdx":785,"cells":{"text":{"kind":"list like","value":[["Pyrazolecarboxamides, intermediates therefor and pest control agent comprising the same as active ingredients","The present invention provides novel compounds with fungicidal and bactericidal activity as well as insecticidal and miticidal activity, which are useful for controlling hazardous biological organisms and belong to pyrazolecarboxamides represented by the following formula (I): wherein R1 represents C1-C5 alkyl; R2 represents C1-C5 alkyl, C1-C5 haloalkyl, or the like; R3 represents hydrogen or C1-C3 alkyl; R4 represents halogen, C1-C5 alkyl or the like; n is an integer of 0 to 4; R5 represents alkyl, haloalkyl, alkoxy, haloalkoxy or the like; n represents an integer of 1 to 5; and X represents hydrogen, C1-C3 alkyl, C1-C3 alkoxy, halogen, C1-C3 haloalkoxy or nitro."],["1.Pyrazolecarboxamide represented by the following formula (I): wherein R1 represents C1-C5 alkyl, R2 represents C1-C5 alkyl, C1-C5 haloalkyl, C1-C5 alkoxy or C1-C5 haloalkoxy, R3 represents hydrogen or C1-C3 alkyl, R4 represents C1-C5 alkyl, C1-C5 haloalkyl, C1-C5 alkoxy, C1-C5 haloalkoxy or halogen, m is an integer of 0 to 4; R5 represents alkyl, haloalkyl, alkoxy, haloalkoxy, alkylthio, haloalkylthio, alkylsulfinyl, haloalkylsulfinyl, alkylsulfonyl, haloalkylsulfonyl, halogen, cyano, formyl, vinyl, alkoxycarbonyl, haloalkoxycarbonyl, acetyl, hydroxycarbonyl, alkylcarbamoyl or a group represented by wherein R7 and R8 each represents hydrogen or C1-C5 alkyl, n represents an integer of 1 to 5, X represents hydrogen, C1-C3 alkyl, C1-C3 alkoxy, halogen, C1-C3 haloalkoxy or nitro, or R2 and X may be taken together to form wherein R9 represents hydrogen or C1-C3 alkyl.","2.The pyrazolecarboxamide according to claim 1, wherein R1 in formula (I) is methyl.","3.The pyrazolecarboxamide according to claim 1, wherein R2 in formula (I) is ethyl.","4.The pyrazolecarboxamide according to claim 1, wherein R5 in formula (I) is C1-C5 haloalkyl or C1-C5 haloalkoxy.","5.The pyrazolecarboxamide according to any one of claims 1 to 4 claim 1, wherein R3 in formula (I) is hydrogen.","6.The pyrazolecarboxamide according to claim 1, wherein X in formula (I) is hydrogen.","7.The pyrazolecarboxamide according to claim 1, wherein, in formula (I), m is an integer of 0 to 2 and n is 1 or 2.8.Phenylbenzylamine represented by the following formula (II): wherein R3 represents hydrogen or C1-C3 alkyl, R4 represents C1-C5 alkyl, C1-C5 haloalkyl, C1-C5 alkoxy, C1-C5 haloalkoxy or halogen, m is an integer of 0 to 4; n represents an integer of 1 to 5; and R6 represents haloalkyl or haloalkoxy, or an acid addition salt thereof.","9.A pest control agent comprising the pyrazolecarboxamide according to claim 1 as an active ingredient.","10.A fungicidal and bactericidal agent comprising the pyrazolecarboxamide according to claim 1 as an active ingredient.","11.An insecticidal and miticidal agent comprising the pyrazolecarboxamide according to claim 1 as an active ingredient thereof."],["<SOH> BACKGROUND OF THE INVENTION <EOH>In the field of agriculture as well as horticulture and floriculture, various fungicides and bactericides, insecticides and miticides have been developed and used in a practical sense for the purpose of controlling various hazardous insects and plant diseases.","However, it cannot be said that agricultural chemicals for common use in the related art have been satisfactory in terms of, for example, effect, spectrum and residual effect or have satisfied the demand to reduce the number and chemical dose to be applied.","Additionally, the emergence of hazardous insects and plant diseases with acquired resistance against agricultural chemicals for common use in the related art is now problematic.","For the cultivation of vegetables, fruit trees, flowers, teas, wheat, barley, oats, etc., rice plants and the like, various hazardous insects with acquired resistance against diverse types of insecticides of, for example, carbamate-series, pyrethroid-series, benzoyl urea-series, organic chloride-series, and organic phosphorus-series and various pathological microorganisms and bacteria with acquired resistance against various types of fungicides and bactericides of, for example, triazole-series, imidazole-series, pyrimidine-series, benzimidazole-series, dicarboximide-series, and phenylamide-series have emerged in various local regions.","Therefore, it has been very tough increasingly year by year to control these various hazardous insects and pathological microorganisms and bacteria causing plant diseases, because of such emergence of these resistant hazardous insects.","Thus, it has been consistently desired the development of a novel agricultural chemical of which a lower chemical dose can express a sufficient effect on the control of various pathological microorganisms and bacteria and hazardous insects with acquired resistance against insecticides and fungicides and bactericides for agriculture and horticulture and floriculture in the related art and which shows less adverse effects on environment.","The present inventors found that N-benzylpyrazole-5-carboxamide derivatives had insecticidal and miticidal activity.","Specifically, the publication of JP-A-64-25763 discloses the following compound with insecticidal and miticidal activity, and the like.","Additionally, the publication of JP-A-3-81266 discloses that the following compound with phenoxy and the like have insecticidal and miticidal activity.","However, Compound (III) with a structure similar to the compound of the present invention as disclosed in JP-A-64-25763 is so insufficient in view of the insecticidal and miticidal activity that the practical applicability of the compound is still low.","Meanwhile, a compound with both fungicidal and bactericidal activity and insecticidal and miticidal activity is very useful for use in agriculture and horticulture and floriculture.","Regarding fungicidal and bactericidal activity, it is known that only a compound with specific substituents among compounds described in JP-A-64-25763 has fungicidal and bactericidal activity (JP-A-3-206079).","An object of the present invention to provide a new substance with a great effect on the control of various pathological microorganisms and bacteria and with usefulness in the control of insects and mites, particularly a highly safe substance with a great effect on the control of various hazardous insects and pathological bacteria causing plant diseases with acquired resistance against fungicides and bactericides and insecticides in the related art, because such a lower chemical dose thereof can express these effects that problems of residual toxicity and environmental pollution can be reduced."],["TECHNICAL FIELD The present invention relates to novel pyrazolecarboxamides and intermediates therefor, and pest control agents comprising the same as an active ingredient.","The novel pyrazolecarboxamides in accordance with the present invention are useful as bactericides and fungicides, insecticides and miticides for agriculture, and horticulture and floriculture.","BACKGROUND OF THE INVENTION In the field of agriculture as well as horticulture and floriculture, various fungicides and bactericides, insecticides and miticides have been developed and used in a practical sense for the purpose of controlling various hazardous insects and plant diseases.","However, it cannot be said that agricultural chemicals for common use in the related art have been satisfactory in terms of, for example, effect, spectrum and residual effect or have satisfied the demand to reduce the number and chemical dose to be applied.","Additionally, the emergence of hazardous insects and plant diseases with acquired resistance against agricultural chemicals for common use in the related art is now problematic.","For the cultivation of vegetables, fruit trees, flowers, teas, wheat, barley, oats, etc., rice plants and the like, various hazardous insects with acquired resistance against diverse types of insecticides of, for example, carbamate-series, pyrethroid-series, benzoyl urea-series, organic chloride-series, and organic phosphorus-series and various pathological microorganisms and bacteria with acquired resistance against various types of fungicides and bactericides of, for example, triazole-series, imidazole-series, pyrimidine-series, benzimidazole-series, dicarboximide-series, and phenylamide-series have emerged in various local regions.","Therefore, it has been very tough increasingly year by year to control these various hazardous insects and pathological microorganisms and bacteria causing plant diseases, because of such emergence of these resistant hazardous insects.","Thus, it has been consistently desired the development of a novel agricultural chemical of which a lower chemical dose can express a sufficient effect on the control of various pathological microorganisms and bacteria and hazardous insects with acquired resistance against insecticides and fungicides and bactericides for agriculture and horticulture and floriculture in the related art and which shows less adverse effects on environment.","The present inventors found that N-benzylpyrazole-5-carboxamide derivatives had insecticidal and miticidal activity.","Specifically, the publication of JP-A-64-25763 discloses the following compound with insecticidal and miticidal activity, and the like.","Additionally, the publication of JP-A-3-81266 discloses that the following compound with phenoxy and the like have insecticidal and miticidal activity.","However, Compound (III) with a structure similar to the compound of the present invention as disclosed in JP-A-64-25763 is so insufficient in view of the insecticidal and miticidal activity that the practical applicability of the compound is still low.","Meanwhile, a compound with both fungicidal and bactericidal activity and insecticidal and miticidal activity is very useful for use in agriculture and horticulture and floriculture.","Regarding fungicidal and bactericidal activity, it is known that only a compound with specific substituents among compounds described in JP-A-64-25763 has fungicidal and bactericidal activity (JP-A-3-206079).","An object of the present invention to provide a new substance with a great effect on the control of various pathological microorganisms and bacteria and with usefulness in the control of insects and mites, particularly a highly safe substance with a great effect on the control of various hazardous insects and pathological bacteria causing plant diseases with acquired resistance against fungicides and bactericides and insecticides in the related art, because such a lower chemical dose thereof can express these effects that problems of residual toxicity and environmental pollution can be reduced.","DISCLOSURE OF THE INVENTION The inventors have investigated the problems described above.","Consequently, the inventors have found that novel pyrazolecarboxamides with a biphenyl backbone with substituents and with a specific combination of substituents have fungicidal and bactericidal activity and insecticidal and miticidal activity meeting the features described above.","Thus, the present invention has been achieved.","The present invention relates to a pyrazolecarboxamide represented by the following formula (I): wherein R1 represents C1-C5 alkyl, R2 represents C1-C5 alkyl, C1-C5 haloalkyl, C1-C5 alkoxy or C1-C5 haloalkoxy, R3 represents hydrogen or C1-C3 alkyl, R4 represents C1-C5 alkyl, C1-C5 haloalkyl, C1-C5 alkoxy, C1-C5 haloalkoxy or halogen, m is an integer of 0 to 4; R5 represents alkyl, haloalkyl, alkoxy, haloalkoxy, alkylthio, haloalkylthio, alkylsulfinyl, haloalkylsulfinyl, alkylsulfonyl, haloalkylsulfonyl, halogen, cyano, formyl, vinyl, alkoxycarbonyl, baloalkoxycarbonyl, acetyl, hydroxycarbonyl, alkylcarbamoyl or a group represented by wherein R7 and R8 each represents hydrogen or C1-C5 alkyl, n represents an integer of 1 to 5, X represents hydrogen, C1-C3 alkyl, C1-C3 alkoxy, halogen, C1-C3 haloalkoxy or nitro, or R2 and X may be taken together to form wherein R9 represents hydrogen or C3-C3 alkyl, a pest control agents comprising the pyrazolecarboxamide as an active ingredient, a phenylbenzylamine as an intermediate for the pyrazolecarboxamide, which is represented by the following formula (II): wherein R3, R4, m and n have the same meanings as defined in formula (I); R represents haloalkyl or haloalkoxy, and an acid addition salt thereof.","BEST MODE FOR CARRYING OUT THE INVENTION The present invention is now described in detail below.","Pyrazolecarboxamides: In the pyrazolecarboxamides represented by formula (I) in accordance with the present invention, the substituent R1 represents a linear or branched C1-C5 alkyl such as methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, 2-methylbutyl, 3-methylbutyl, isopentyl, 2-ethylpropyl, neopentyl, 2,2-dimethylpropyl, or 2-methylisobutyl.","Among these, methyl is preferable as the substituent R1.The substituent R2 represents linear, branched or cyclic C1-C5 alkyl such as methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, 2-methylbutyl, 3-methylbutyl, isopentyl, 2-ethylpropyl, neopentyl, 2,2-dimethylpropyl, 2-methylisobutyl, or cyclopropyl; linear or branched C1-C5 haloalkyl such as fluoronethyl, chloromethyl, bromomethyl, difluoromethyl, difluoroethyl, trifluoromethyl, trichloromethyl, tribromomethyl, 2-fluoroethyl, 2,2,2-trifluoroethyl, 1-(trifluoromethyl)ethyl, 1,1,2,3,3,3-hexafluoropropyl, 2,2,3,3,3-pentafluoropropyl, 2,2,3,3-tetratluoropropyl, 2,2,3,3,4,4,4-heptafluorobutyl, or 2,2,3,3,4,4,5,5,5-nonafluoropentyl; linear or branched C1-C5 alkoxy such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, isobutoxy, tert-butoxy, n-pentoxy, 2-methylbutoxy, 3-methylbutoxy, isopentoxy, 2-ethylpropoxy, neopentoxy, 2,2-dimethylpropoxy, or 2-methylisobutoxy; or linear or branched C1-C5 haloalkoxy such as difluoromethoxy, trifluoromethoxy, fluoromethoxy, trichloromethoxy, tribromomethoxy, 2-fluoroethoxy, 2,2,2-trifluoroethoxy, 2,2,2-trichloroethoxy, 1-(trifluoromethyl)ethoxy, 1,1,2,3,3,3-hexafluoropropoxy, 2,2,3,3,3-pentafluoropropoxy, or 2,2,3,3-tetrafluoropropoxy.","Among these, linear C1-C3 alkyl is preferable as the substituent R2 Ethyl is particularly preferable.","The substituent R3 represents hydrogen; or linear or branched C1-C3 alkyl such as methyl, ethyl, n-propyl, or isopropyl.","Among these, hydrogen is preferable as the substituent R3.The substituent R4 represents linear or branched C1-C5 alkyl such as methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, 2-methylbutyl, 3-methylbutyl, isopentyl, 2-ethylpropyl, neopentyl, 2,2-dimethylpropyl, or 2-methylisobutyl; linear or branched C1-C5 haloalkyl such as fluoromethyl, chloromethyl, bromomethyl, difluoromethyl, difluoroethyl, trifluoromethyl, trichloromethyl, tribromomethyl, 2-fluoroethyl, 2,2,2-trifluoroethyl, 1-(trifluoromethyl)ethyl, 1,1,2,3,3,3-hexafluoropropyl, 2,2,3,3,3-pentafluoropropyl, 2,2,3,3-tetrafluoropropyl, 2,2,3,3,4,4,4-heptafluorobutyl, or 2,2,3,3,4,4,5,5,5-nonafluoropentyl; linear or branched C1-C5 alkoxy such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, isobutoxy, text-butoxy, n-pentoxy, 2-methylbutoxy, 3-methylbutoxy, isopentoxy, 2-ethylpropoxy, neopentoxy, 2,2-dimethylpropoxy, or 2-methylisobutoxy; linear or branched C1-C5 haloalkoxy such as difluoromethoxy, trifluoromethoxy, fluoromethoxy, trichloromethoxy, tribromomethoxy, 2-fluoroethoxy, 2,2,2-trifluoroethoxy, 2,2,2-trichloroethoxy, 1-(trifluoromethyl)ethoxy, 1,1,2,3,3,3-hexafluoropropoxy, 2,2,3,3,3-pentafluoropropoxy, or 2,2,3,3-tetrafluoropropoxy; or halogen such as fluorine, chlorine, bromine or iodine.","Among these, halogen is preferable as the substituent R4.The substituent R5 represents linear or branched alkyl such as methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, 2-methylbutyl, 3-methylbutyl, isopentyl, 2-ethylpropyl, neopentyl, 2,2-dimethylpropyl, 2-methylisobutyl, n-hexyl, n-heptyl or n-octyl; linear or branched haloalkyls such as fluoromethyl, chloromethyl, bromomethyl, difluoromethyl, difluoroethyl, trifluoromethyl, trichloromethyl, tribromomethyl, 2-fluoroethyl, 2,2,2-trifluoroethyl, 1-(trifluoromethyl)ethyl, 1,1,2,3,3,3-hexafluoropropyl, 2,2,3,3,3-pentafluoropropyl, 2,2,3,3-tetrafluoropropyl, 2,2,3,3,4,4,4-heptafluoropropyl, or 2,2,3,3,4,4,5,5,5-nonafluoropentyl; linear or branched alkoxys such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, isobutoxy, tert-butoxy, n-pentoxy, 2-methylbutoxy, 3-methylbutoxy, isopentoxy, 2-ethylpropoxy, neopentoxy, 2,2-dimethylpropoxy, 2-methylisobutoxy, n-hexyloxy, n-heptyloxy, or n-octyloxy; linear or branched haloalkoxy such as difluoromethoxy, trifluoromethoxy, fluoromethoxy, trichloromethoxy, tribromoinethoxy, 2-fluoroethoxy, 2,2,2-trifluoroethoxy, 2,2,2-trichloroethoxy, 1-(trifluoromethyl)ethoxy, 1,1,2,3,3,3-hexafluoropropoxy, 2,2,3,3,3-pentafluoropropoxy, 2,2,3,3-tetrafluoropropoxy, or 2,2,3,3,4,4,4-heptafluorobutoxy; linear or branched alkylthio such as methylthio, ethylthio, propylthio, isopropylthio, butylthio, sec-butylthio, isobutylthio, tert-butylthio, pentylthio, 2-methylbutylthio, 3-methylbutylthio, isopentylthio, 2-ethylpropylthio, neopentylthio, 2,2-direthylpropylthio, or 2-methylisobutylthio; haloalkylthio such as trifluoronethylthio, perfluoroethylthio, perfluoropropylthio, or 2,2,2-trifluoroethylthio; linear or branched alkylsulfinyl such as methylsulfinyl, ethylsulfinyl, propylsulfinyl, isopropylsulfinyl, butylsulfinyl, sec-butylsulfinyl, isobutylsulfinyl, tert-butylsulfinyl, pentylsulfinyl, 2-methylbutylsulfinyl, 3-methylbutylsulfinyl, isopentylsulfinyl, 2-ethylpropylsulfinyl, neopentylsulfinyl, 2,2-dimethylpropylsulfinyl or 2-methylisobutylsulfinyl; haloalkylsulfinyl such as trifluoromethylsulfinyl or 2,2,2-trifluoroethylsulfinyl; linear or branched alkylsulfonyl such as methylsulfonyl, ethylsulfonyl, propylsulfonyl, isopropylsulfonyl, butylsulfonyl, sec-butylsulfonyl, isobutylsulfonyl, text-butylsulfonyl, pentylsul fonyl, 2-methylbutylsulfonyl, 3-methylbutylsulfonyl, isopentylsulfonyl, 2-ethylpropylsulfonyl, neopentylsulfonyl, 2,2-dimethylpropylsulfonyl, or 2-methylisobutylsulfonyl; haloalkylsulfonyl such as trifluorosulfonyl or 2,2,2-trifluoroethylsulfonyl; halogen such as fluorine, chlorine, bromine or iodine; cyano; formyl; vinyl; linear or branched alkoxycarbonyl such as methoxycarbonyl, ethoxycarbonyl, n-propoxycarbonyl, isopropoxycarbonyl or butoxycarbonyl; haloalkoxycarbonyl such as 2,2,2-trifluoromethoxycarbonyl or 2,2,3,3,3-pentafluorpropoxycarbonyl; acetyl; hydroxycarbonyl; alkylcarbamoyl such as methylcarbamoyl, ethylcarbamoyl or propylcarbamoyl; or a group represented by Herein, the above R7 and R8 each represents hydrogen; or linear or branched C1-C5 alkyl such as methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, or pentyl.","In the above R5, the alkyl and alkoxy have preferably C1-C8; the haloalkyl, haloalkoxy, alkylthio, haloalkylthio, alkylsulfinyl, haloalkylsulfinyl, alkylsulfonyl and haloalkylsulfonyl have preferably C1-C5.The alkoxycarbonyl, haloalkoxycarbonyls, alkylcarbamoyls and group represented by have preferably C2-C5, more preferably C2-C5.Among these, the substituent R5 is preferably haloalkyl or haloalkoxy, more preferably C1-C5 haloalkyl or C1-C5 haloalkoxy, particularly preferably trifluoromethoxy or difluoromethoxy.","Additionally, the substituent is preferably positioned at the 4-position.","The substituent X represents hydrogen; linear or branched C1-C3 alkyl such as methyl, ethyl, n-propyl, and isopropyl; linear or branched C1-C3 alkoxy such as methoxy, ethoxy, n-propoxy, or isopropoxy; halogen such as fluorine, chlorine, bromine, or iodine; C1-C3 haloalkoxy such as difluoromethoxy, 2,2,2-trifluoroethoxy, or perfluoropropoxy; or nitro.","Among these, the substituent X is preferably hydrogen.","Furthermore, R9 in the group formed by R2 and X together represents hydrogen; or C1-C3 alkyl such as methyl, ethyl, n-propyl or isopropyl.","m is an integer of 0 to 4, preferably 0 to 2; and n is an integer of 1 to 5, preferably 1 or 2.Phenylbenzylamines: Furthermore, the phenylbenzylamines represented by formula (II) and acid addition salts thereof in accordance with the present invention are novel intermediates for producing the pyrazolecarboxamides.","Herein, the substituents R3 and R4, m and n have the same meanings as defined in formula (I).","The substituent R6 represents haloalkyl such as fluoromethyl, chloromethyl, bromomethyl, difluoromethyl, difluoroethyl, trifluoromethyl, trichloromethyl, tribromomethyl, 2-fluoroethyl, 2,2,2-trifluoroethyl, 1-(trifluoromethyl)ethyl, 1,1,2,3,3,3-hexafluoropropyl, 2,2,3,3,3-pentafluoropropyl, 2,2,3,3-tetrafluoropropyl, 2,2,3,3,4,4,4-heptafluorobutyl, or 2,2,3,3,4,9,5,5,5-nonafluoropentyl; or haloalkoxys such as difluoromethoxy, trifluoromethoxy, fluoromethoxy, trichloromethoxy, tribromomethoxy, 2-fluoroethoxy, 2,2,2-trifluoroethoxy, 2,2,2-trichloroethoxy, 1-(trifluoroimethyl)ethoxy, 1,1,2,3,3,3-hexafluoropropoxy, 2,2,3,3,3-pentafluoropropxy, 2,2,3,3-tetrafluoropropoxy, or 2,2,3,3,4,4,4-heptafluorobutoxy.","The haloalkyl and haloalkoxy have preferably C1-C5.The acid to be added to the phenylbenzylamines represented by formula (II) in accordance with the present invention includes carboxylic acid such as acetic acid, propionic acid, butyric acid, oxalic acid, adipic acid, dodecanedioic acid, lauric acid, stearic acid, trifluoroacetic acid, fumaric acid, maleic acid, benzoic acid, or phthalic acid; sulfonate such as methanesulfonic acid, 1,3-propanedisulfonic acid, p-toluenesulfonic acid, or dodecylbenzenesulfonic acid; and inorganic acid such as hydrochloric acid, sulfuric acid, nitric acid, or carbonic acid.","Production Process: The compound represented by formula (I) in accordance with the present invention is a novel compound and can be produced, for example, by the following reaction scheme.","(In the formula, R1, R2, R3, R4, R5, X, m and n have the same meanings as described in formula (I).)","In other words, pyrazolecarboxylic acid chloride (IV) and phenylbenzylamine (V) are allowed to react in the presence or absence of a base, preferably using a solvent at −10 to 50° C., preferably 0 to 25° C. As the solvent, any solvent can be used, so long as it does not have direct influence on the reaction.","The solvent includes aromatic hydrocarbons such as benzene, toluene and xylene; ketones such as acetone, methyl ethyl ketone, and methyl isobutyl ketone; halogenated hydrocarbons such as chloroform and dichloromethane; water; esters such as methyl acetate and ethyl acetate; and polar solvents such as tetrahydrofuran, acetonitrile, dioxane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, and pyridine.","The base includes alkali metal hydrides such as sodium hydride; alkali metal hydroxides such as sodium hydroxide and potassium hydroxide; alkali metal carbonates such as sodium carbonate and potassium carbonate; inorganic salts; and amines such as pyridine and triethylamine.","In order to isolate the objective compound of formula (I) after the reaction, the following procedures are carried out in case of using a solvent soluble in water: distilling off the solvent under reduced pressure, adding water to the resulting residue, extracting the residue in aromatic hydrocarbons insoluble in water such as benzene, toluene and xylene, halogenated hydrocarbons such as chloroform and dichloromethane, and esters such as ethyl acetate, rinsing the extract with a saturated aqueous sodium chloride solution, drying the extract over desiccants such as anhydrous sodium sulfate or anhydrous magnesium sulfate and then distilling off the solvents under reduced pressure.","When a solvent insoluble in water is used, water is added to the reaction mixture, followed by separation; the resulting organic layer is rinsed with a saturated aqueous sodium chloride solution and dried over desiccants such as anhydrous sodium sulfate or magnesium sulfate, from which the solvent is distilled off under reduced pressure.","The residue recovered after distillation of the solvent is purified by recrystallization, suspending and rinsing, and column chromatography, so that the objective compound represented by formula (I) can be recovered.","The compound represented by formula (IV) as one raw material can be synthetically prepared, for example, by the method described in JP-A-64-25763.The compound represented by formula (V) as one raw material can be synthetically prepared, for example, by the following method.","(1) Reduction of Phenylbenzonitriles (In the formula, R4, R5, in and n have the same meanings as described in formula (I).)","Phenylbenzonitrile (VI) and hydrogen are allowed to react in the presence of a metal catalyst and aqueous ammonia, preferably using an organic solvent, at 0 to 100° C., preferably 25 to 50° C. As the organic solvent, any organic solvent can be used, so long as it does not have directly influence on the reaction.","The organic solvent includes alcohols such as methanol, ethanol and isopropanol; aromatic hydrocarbons such as benzene, toluene and xylene; ketones such as acetone, methyl ethyl ketone, and methyl isobutyl ketone; halogenated hydrocarbons such as chloroform and dichloromethane; water; esters such as methyl acetate and ethyl acetate; or polar solvents such as tetrahydrofuran, acetonitrile, dioxane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, and pyridine.","The metal catalyst includes Raney-nickel, palladium carbon, and platinum oxide.","In order to isolate the objective compound of formula (Va) after the reaction, the metal catalyst is filtered off from the reaction mixture, and the solvent is distilled off from the resulting filtrate under reduced pressure.","The resulting product may be sufficiently pure as it is, but the product can be purified by means of evaporation and column chromatography, to recover a pure product.","Phenylbenzonitrile (VI) can be synthetically prepared by reaction of the corresponding haloallene with phenylboronic acid in the presence of a zero-valent palladium catalyst and a base according to known methods, for example, the method described in Synthetic Communications, Vol.","11, page 513 (1981).","(2) Reduction of Oximes Oxime (VII) and hydrogen are allowed to react in the presence of a metal catalyst and aqueous ammonia, preferably using an organic solvent, at 0 to 100° C., preferably 25 to 50° C. As the organic solvent, any organic solvent can be used, so long as it has not direct influence on the reaction.","The organic solvent includes alcohols such as methanol, ethanol and isopropanol; aromatic hydrocarbons such as benzene, toluene and xylene; ketones such as acetone, methyl ethyl ketone, and methyl isobutyl ketone; halogenated hydrocarbons such as chloroform and dichlioromethane; water; esters such as methyl acetate and ethyl acetate; and polar solvents such as tetrahydrofuran, acetonitrile, dioxane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, and pyridine.","The metal catalyst includes Raney-nickel, palladium carbon, and platinum oxide.","In order to isolate the objective compound of formula (Vb) after the reaction, the metal catalyst is filtered off from the reaction mixture, and the solvent is distilled off from the resulting filtrate under reduced pressure.","Oxime (VII) can be synthetically prepared by reaction of the corresponding acetophenone with hydroxyamine in the presence of potassium carbonate according to known methods.","Phenylbenzylamine-acid addition salts (Vc) can be synthesized by the following method.","Phenylbenzylamine (V) and acid (Y) are allowed to react, preferably using a solvent, at −5 to 50° C., preferably 0 to 25° C. As the acid, organic acid and inorganic acid can be used.","The organic acid includes carboxylic acids such as acetic acid, propionic acid, butyric acid, oxalic acid, adipic acid, dodecanedioic acid, lauric acid, stearic acid, trifluoroacetic acid, fumaric acid, maleic acid, benzoic acid, and phthalic acid; and sulfonates such as methanesulfonic acid, 1,3-propanedisulfonic acid, p-toluenesulfonic acid, and dodecylbenzenesulfonic acid.","The inorganic acid includes hydrochloric acid, sulfuric acid, nitric acid and carbonic acid.","As the solvent, any solvent can be used, so long as it does not have direct influence on the reaction.","The organic solvent includes aromatic hydrocarbons such as benzene, toluene and xylene; ketones such as acetone, methyl ethyl ketone, and methyl isobutyl ketone; halogenated hydrocarbons such as chloroform and dichloromethane; water; esters such as methyl acetate and ethyl acetate; and polar solvents such as tetrahydrofuran, acetonitrile, dioxane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, and pyridine.","After the reaction, the solvent is distilled off under reduced pressure.","The resulting crystals are filtered and recovered, to synthetically prepare inorganic acid salts of benzylamine and organic acid salts thereof.","The crystals are satisfactory as they are.","For further purification, however, the crystals are rinsed with the organic solvents described above, to synthetically prepare benzylamine salts at high purity.","Pest Control Agents: The compound represented by formula (I) in accordance with the present invention has high fungicidal and bactericidal effects on pathological microorganisms and bacteria against plants such as Pyricularia oxyzae, pathological microorganisms and bacteria causing powdery mildew, pathological microorganisms and bacteria causing rust, and pathological microorganisms and bacteria causing downy mildew so that the compound is useful as an active ingredient for fungicides and bactericides for agriculture and horticulture and floriculture.","Since the compound of the present invention has high activity on the control of the eggs and larvae of insects of the order Hemiptera including leafhoppers such as Sogatella furcifera Horvath, Nilaparvata lugens (Stal), and Laodelphax stratella (Fallen); Cicadomorpha-2 such as Bothrogonia ferruginea (Fabricius) and Cicadella viridis (Linnaeus); and Aphididae such as Myzus persicae; the order Lepidoptera including Spodoptera litura Fabricius, Chilo suppresalis, Cnaphalocrosis medinalis Guenee and Plutella xylostella (Linnaeus); the order Coleoptera including Callosobruchus chinensis; the order Diptera including Musca domestica Linnaeus, Aedes aegypti and Culex pipiens pallens; the order Orthoptera; and the order Acarina including Tetranychus urticae Koch, Tetranychus cinnabarinus (Boisduval) and Panonychus citri (Mcgregor), the compound of the present invention is useful as an active ingredient for insecticides and miticides for agriculture and horticulture and floriculture.","It is needless to say that the pathological microorganisms and bacteria against plants, insects, and mites as subjects to be controlled with the compound of the present invention are not limited to those described above.","When the compound represented by formula (I) in accordance with the present invention is used for fungicides and bactericides, insecticides and/or miticides for agriculture and horticulture and floriculture, the compound may satisfactorily be used singly.","However, preferably, the compound may be used in composition forms produced using agricultural auxiliary agents for common use in the art, such as fungicides and bactericides, insecticides and/or miticide's for agriculture and horticulture and floriculture.","The forms are preferably, for example, emulsions, hydrates, powders, flowable agents, fine granules, granules, tablets, oils, sprays, and fumes.","However, the forms are not limited thereto.","One or two or more forms of the compound may also be blended as active ingredients.","The agricultural auxiliary agents for use in producing the fungicides and bactericides, insecticides and/or miticides for agriculture and horticulture and floriculture are used, for example, for the improvement of the effects of fungicides and bactericides, insecticides and/or miticides for agriculture and horticulture and floriculture and for the improvement of stability and dispersibility.","For example, carriers (diluents), spreading agents, emulsifiers, wet spreading agents, dispersants and disintegrators can be used.","The liquid carriers include water, aromatic hydrocarbons such as toluene and xylene, alcohols such as methanol, butanol, and glycol, ketones such as acetone, amides such as dimethylformamide, sulfoxides such as dimethyl sulfoxide, methylnaphthalene, cyclohexane, animal and vegetable oils, and fatty acid.","Additionally, the solid carriers include clay, kaolin, talc, diatomaceous earth, silica, calcium carbonate, montmollionite, bentonite, feldspar, quartz, alumina, sawdust, nitrocellulose, starch and gum arabic.","As the emulsifiers and the dispersants, general surfactants can be used and include anionic surfactants, cationic surfactants, nonionic surfactants and ampholytic surfactants, such as higher alcohol sodium sulfate, stearyltrimethylammonium chloride, polyoxyethylene alkylphenyl ether, and lauryl betaine.","Additionally, the following agents can also be used: spreading agents such as polyoxyethylene nonyl phenyl ether and polyoxyethylene lauryl phenyl ether; wet spreading agents such as dialkylsulfosuccinate; attaching agents such as carboxymethyl cellulose and polyvinyl alcohol; and disintegrators such as sodium lignin sulfonate and sodium lauryl sulfate.","The content of the active ingredient in the fungicides and bactericides, insecticides and/or miticides for agriculture and horticulture and floriculture in accordance with the present invention is selected within a range of 0.1 to 99.5% and is appropriately determined on the basis of various conditions for preparations and application methods.","Preferably, for example, powders are produced so as to contain the active ingredient at about 00.5 to 20% by weight, preferably about 10% by weight; hydrates are produced so as to contain the active ingredient at about 1 to 90% by weight, preferably 10 to 80% by weight; emulsions are produced so as to contain the active ingredient at about 1 to 90% by weight, preferably about 10 to 40% by weight.","In the case of the emulsions, for example, solvents, surfactants and the like are mixed with the compound as the active ingredient to prepare a stock emulsion.","The stock emulsion is diluted with water to a given concentration when used, and is applied.","The resulting solution can be applied.","In the case of the hydrates, the compound as the active ingredient, a solid carrier, a surfactant and the like are mixed together to prepare a stock solution.","The stock solution is diluted with water when used, and is applied.","In the case of the powders, the compound as the active ingredient, a solid carrier and the like are mixed together.","The resulting mixture can be used as it is.","In the case of the granules, the compound as the active ingredient, a solid carrier, a surfactant and the like are mixed together and granulated, to prepare granules, which can be used as it is.","It is needless to say that the methods for preparing the preparations are not limited to those described above.","One skilled in the art can select an appropriate method, depending on the type of the active ingredient, the purpose of application and the like.","In addition to the compound as the active ingredient, the fungicides and bactericides, the insecticides and/or miticides for agriculture and horticulture and floriculture in accordance with the present invention may contain any appropriate active ingredients such as other fungicides and bactericides, insecticides, miticides, herbicides, insect growth controlling agents, fertilizers, and soil modifiers.","The method for applying the fungicides and bactericides, insecticides and/or miticides for agriculture and horticulture and floriculture in accordance with the present invention is not particularly limited, and spraying on stem and leaf, application on water surface, soil treatment, and seed treatment can be carried out.","In the case of spraying on stem and leaf, for example, a solution within a concentration range of 5 to 1,000 ppm, preferably 10 to 500 ppm can be used at a volume of about 100 to 200 liters per 10 Ares.","For application on water surface, generally, granules at 5-15% content of the active ingredient are applied at a ratio of 1 to 10 kg per 10 ares.","In the case of soil treatment, a solution within a concentration range of 5 to 1,000 ppm is applied at a ratio of about 1 to 10 liters per 2, In the case of seed treatment, a solution within a concentration range of 10 to 1,000 ppm is applied at a ratio of about 10 to 100 ml per kg of seeds.","The present invention is now described in the following examples in more detail.","However, the scope of the present invention is not limited to the following examples.","EXAMPLES The present invention is now described below in detail in the following examples, Formulation Examples and test examples.","However, the present invention is not limited to the examples, unless the examples are departed from the scope of the present invention.","Example 1 N-[4-(4-Difluoromethoxyphenyl)benzyl]-1-methyl-3-ethylpyrazole-5-carboxamide: A mixture of 4-(4-difluoromethoxyphenyl)benzylamine (13.8 g), triethylamine (10 ml) and dichloromethane (100 ml) was cooled to 0° C. to 4° C., followed by addition of a solution (20 ml) of 1-methyl-3-ethylpyrazole-5-carboxylic acid chloride (12.4 g) in dichloromethane, for agitation for 30 minutes.","The reaction mixture was poured into water (200 ml), for extraction in dichloromethane.","The organic layer was rinsed with a saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfate.","The solvent was distilled off under reduced pressure.","The resulting residue was purified by silica gel chromatography, to obtain 17.0 g of Compound No.","30 described in Table-1.The melting point was 107 to 108° C. Example 2 According to the method in Example 1, the compounds described in Table-1 were prepared synthetically.","TABLE 1 Compound No.","R1 R2 X R3 m.p.","(° C.) or nD/(° C.) 1 CH3 CH3 CH3 H 162-163 2 CH3 CH3 C2H5O H 1.5464/23.0 3 CH3 CH3 H H 128-130 4 CH3 CH3 H H 98-99 5 CH3 CH3 H H 134-135 6 CH3 CH3 H H 127-128 7 CH3 CH3 H H 122-124 8 CH3 C2H5 H H 116-118 9 CH3 C2H5 H H 119-121 10 CH3 C2H5 H H 106-107 11 CH3 C2H5 H H 104-108 12 CH3 C2H5 H H 105-107 13 CH3 C2H5 H H 110-111 14 CH3 C2H5 H H 90-91 15 CH3 C2H5 H H 95-97 16 CH3 C2H5 H H 1.5671/23.0 17 CH3 C2H5 H H 143-145 18 CH3 C2H5 Cl H 124-125 19 CH3 C2H5 H H 132-133 20 CH3 C2H5 Cl H 142-143 21 CH3 C2H5 H H 143-144 22 CH3 C2H5 Cl H 155-156 23 CH3 C2H5 H H 151-153 24 CH3 C2H5 H H 124-126 25 CH3 C2H5 H H 122-123 26 CH3 C2H5 H H 126-127 27 CH3 C2H5 H H 122-123 28 CH3 C2H5 H H 96-97 29 CH3 C2H5 H H 91-93 30 CH3 C2H5 H H 107-108 31 CH3 C2H5 F H 124-125 32 CH3 C2H5 H H 1.5623/25.0 33 CH3 C2H5 H H 109-110 34 CH3 C2H5 H CH3 81-82 35 CH3 C2H5 F H 105-106 36 CH3 C2H5 Cl H 93-95 37 CH3 C2H5 H H 114-116 38 CH3 C2H5 Cl H 117-118 39 CH3 C2H5 H H 1.5523/23.0 40 CH3 C2H5 Cl H 96-97 41 CH3 C2H5 H H 95-96 42 CH3 C2H5 Cl H 106-107 43 CH3 C2H5 H H 95-96 44 CH3 C2H5 H CH3 122-123 45 CH3 C2H5 H H 99-100 46 CH3 C2H5 Cl H 108-109 47 CH3 C2H5 H H 113-114 48 CH3 C2H5 Cl H 106-107 49 CH3 C2H5 H H 118-119 50 CH3 C2H5 H H 163-164 51 CH3 C2H5 Cl H 136-137 52 CH3 C2H5 H H 119-121 53 CH3 C3H5 H H 141-143 54 CH3 C2H5 H H 154-156 55 CH3 C2H5 H CH3 147-150 56 CH3 C2H5 H H 1.5865/25.0 57 CH3 i-C3H7 Br H 142-143 58 CH3 CF3 H H 153-154 59 C2H5 C2H5 H H 127-128 60 i-C3H7 C2H5 H H 1.5520/25.0 61 CH3 C2H5 H H 1.5530/23.0 62 CH3 C3H5 H H 1.5671/25.0 63 CH3 C2H5 H H 81-83 64 CH3 C2H5 F H 101-104 65 CH3 C2H5 H H 87-89 66 CH3 C2H5 H H 1.5476/23.0 67 CH3 C2H5 H H 123-124 68 CH3 CH3 H H 111-112 69 CH3 i-C3H7 H H 120-122 70 CH3 H H 99-100 71 CH3 CH3 H H 88-89 72 CH3 C2H5 Cl H 106-107 73 CH3 C2H5 H H 117-118 74 CH3 C2H5 H H 155-157 75 CH3 C2H5 H H 130-131 76 CH3 C2H5 H H 167-168 77 CH3 C2H5 H H 245-247 78 CH3 C2H5 H H 140-141 79 CH3 C2H5 Cl H 154-156 80 CH3 C2H5 H H 102-104 81 CH3 CH3 H H 128-129 82 CH3 C2H5 Cl H 122-123 83 CH3 C2H5 H H 155-160 84 CH3 n-C3H7 H H 131-135 85 CH3 t-C4H9 H H 123-125 86 CH3 CH3 CH3O H 75-77 87 CH3 CH3 n-C3H7O H 51-54 88 CH3 CH3 CHF2O H 136-138 89 CH3 C2H5 H H 209-211 90 CH3 C2H5 H H 139-141 91 CH3 C2H5 H H 242-245 92 CH3 C2H5 H H 162-164 93 CH3 C2H5 H H 152-154 94 CH3 C2H5 H H 146-147 95 CH3 C2H5 H H 131-136 96 CH3 CH3O H H 124-125 97 CH3 C2H5O H H 104-105 98 CH3 i-C3H7O H H 1.5480/25.0 99 CH3 CHF2O H H 137-139 100 CH3 CH3 Cl H 117-118 101 CH3 n-C3H7O H H 81-83 102 CH3 CH3 NO2 H 198-199 103 CH3 H H H 123-125 104 CH3 H 155-156 105 CH3 H 157-159 106 CH3 H 194-195 107 CH3 H 190-192 108 CH3 CH3 H H 150-151 Example 3 Synthesis of 4-(4-difluoromethoxyphenyl)benzylamine: A mixture of 4-cyano-4′-difluoromethoxybiphenyl (15.0 g), Raney-nickel R-100 (15 g) and aqueous 28% ammonia (15.0 ml) was shaken in hydrogen atmosphere at 50° C. for 4 hours.","After the reaction mixture was cooled to ambient temperature, Raney-nickel was filtered off.","The solvent was distilled off under reduced pressure, to obtain 13.8 g of Compound No.","113 described in Table-2.The melting point was 109 to 11° C. Example 4 Synthesis of 4-(4-trifluoromethoxy)-α-methylbenzylamine: A mixture of 4-(4-trifluoromethoxyphenyl)acetophenone oxime (8.8 g), Raney-nickel R-100 (10 g) and aqueous 28% ammonia (10.0 ml) was shaken in hydrogen atmosphere at 50° C. for 3 hours.","After the reaction mixture was cooled to ambient temperature, Raney-nickel was filtered off.","The solvent was distilled off under reduced pressure, to obtain 7.61 g of Compound No.","117 described in Table-2.The melting point was 70 to 74° C. Example 5 Synthesis of 4-(4-trifluoromethoxyphenyl)benzylamine Hydrochloride: To a solution (20 ml) of 4-(4-trifluoromethoxyphenyl)benzylamine (5 g) in methanol, 12N hydrochloric acid (3.0 ml) was added, followed by agitation at ambient temperature for 2 hours, The solvent was distilled off under reduced pressure.","The resulting residue was rinsed with methanol, to obtain 4.8 g of Compound No.","116 described in Table-2.The melting point was 252 to 254° C. Example 6 According to the methods described in Examples 3 to 5, the compounds described in Table-2 were synthetically prepared.","TABLE 2 Compound No.","R3 Y m.p.","(° C.) or nD/(° C.) 109 H — 1.5459/23.0 110 H — 1.5530/23.0 111 H — 143-145 112 H — 109-110 113 H — 109-110 114 H HCl 220-225 115 H — 80-82 116 H HCl 252-254 117 CH3 — 70-74 118 H — 1.5430/23.0 119 H — 1.5323/23.0 120 H — 1.4911/23.0 121 H HCl 176-180 122 H — 1.5488/23.0 123 H — 1.5030/23.0 124 H — 1.5350/23.0 125 H HCl 302-303 126 H — 1.5540/23.0 127 H — 87-89 128 H — 1.5140/25.0 129 H — 1.5531/25.0 130 H — 1.5380/23.0 131 H — 1.5273/25.0 132 H — 1.5222/25.0 133 H — 1.5301/23.0 *Compound No.130 b.p.","= 134-135° C./1 mmHg Hereinbelow, Formulation Examples containing the compound of the present invention as the active ingredient are described, which are insecticides and miticides for agriculture and horticulture and floriculture.","However, the mode for carrying out the present invention is not limited to those described below.","Formulation Example 1 Hydrates: Twenty parts by weight of the compound of the present invention, 20 parts by weight of Carplex #80 (trade name; white carbon; manufactured by Shionogi & Co., Ltd.), 52 parts by weight of ST kaolin clay (trade name; kaolinite; manufactured by Tuchiya Kaolin Co., Ltd.), 5 parts by weight of Solpol 9047K (trade name; anionic surfactant; manufactured by Toho Chemical, Co., Ltd.) and 3 parts by weight of Lunox P56L (trade name; anionic surfactant; Toho Chemical Co., Ltd.) were blended together, uniformly mixed together and ground, to obtain hydrates containing the active ingredient at a content of 20% by weight.","Formulation Example 2 Powders; Two parts by weight of the compound of the present invention, 93 parts by weight of clay (manufactured by Nippon Talc Co., Ltd.), and 5 parts by weight of Carplex #80 (trade name; white carbon; manufactured by Shionogi Pharmaceutical Co., Ltd.) were uniformly mixed together and ground, to obtain powders containing the active ingredient at a content of 2% by weight.","Formulation Example 3 Emulsions: Twenty parts by weight of the compound of the present invention were dissolved in a mixture solvent of 35 parts by weight of xylene and 30 parts by weight of dimethylformamide, followed by addition of 15 parts by weight of Solpol 3005X (trade name; a mixture of a nonionic surfactant and an anionic surfactant; Toho Chemical Co., Ltd.), to obtain emulsions containing the active ingredient at a content of 20% by weight.","Formulation Example 4 Flowable Agents: Thirty parts by weight of the compound of the present invention, 5 parts by weight of Solpol 9047K, 3 parts by weight of Sorbon T-20 (trade name; nonionic surfactant; Toho Pharmaceutical Co., Ltd.), 8 parts by weight of ethylene glycol and 44 parts by weight of water were ground by wet process with Dinomill (manufactured by Sinmaru Enterprise, Co., Ltd.).","Ten parts by weight of an aqueous 1% by weight solution of xanthan gum (naturally occurring polymer) were added to the resulting slurry mixture, followed by thorough mixing and grinding, to obtain flowable agents containing the active ingredient at a content of 20% by weight.","Test examples of fungicides and bactericides, insecticides and miticides for agriculture and horticulture and floriculture are shown below, which contain the compound of the present invention as the active ingredient.","However, the mode for carrying out the present invention is not limited to those described below.","Test Example 1 Fungicidal and Bactericidal Effect on Wheat Powdery Mildew: Compound (III) disclosed in JP-A-64-25763, tebufenpyrade and various compounds in accordance with the present invention were individually prepared into preparations in the same manner as in Formulation 3 The resulting preparations were diluted to given concentrations and sprayed on the stems and leave of a wheat plant (species: Norin No.61) at the one-leaf to 2-leaf stage as grown in a pot of a 6-cm diameter, at a ratio of 10-ml volume per one pot.","After drying the chemical preparations in air, the wheat plant was exposed to the spore collected from wheat leave afflicted with Erypsiphe graminis as the microorganism causing wheat powdery mildew, for inoculation.","Then, the plant was left to stand alone in a green house for 7 to 10 days.","Evaluation was carried out follows.","The ratio of the diseased area of each leaf was examined to calculate the control ratio according to the following formula.","The results are shown in Table 3 (in the table, Compound Nos.","correspond to Compound Nos.","in Table-1.)","Control ratio (%) =[(mean ratio of diseased areas in non-treated lot) −(mean ratio of diseased areas in treated lot)]× ×100 / (mean ratio of diseased areas in non-treated lot) TABLE 3 Compound No.","Concentration (ppm) Control Ratio (%) 3 500 100 4 500 100 5 500 100 6 500 100 7 500 100 13 500 100 17 500 100 21 500 100 25 500 100 30 500 100 31 500 100 34 500 100 35 500 100 36 500 100 37 500 100 39 500 100 40 500 100 41 500 100 42 500 100 45 500 100 46 500 100 47 500 100 48 500 100 49 500 100 54 500 100 63 500 100 64 500 100 68 500 100 69 500 100 70 500 100 71 500 100 72 500 100 78 500 100 80 500 100 81 500 100 82 500 100 84 500 100 85 500 100 86 500 100 87 500 100 88 500 100 90 500 100 91 500 100 92 500 100 98 500 100 101 500 100 (III) 500 0 tebufenpyrade 500 77 Test Example 2 Fungicidal and Bactericidal Effect on Wheat Rust: Compound (III) disclosed in JP-A-64-25763 and various compounds in accordance with the present invention were individually prepared into preparations in the same manner as in Preparation 3.The resulting preparations were diluted to given concentrations and sprayed on the stems and leave of a wheat seedling (species: Norin No.61) at the one-leaf to 2-leaf stage as grown in a pot of a 6-cm diameter, at a ratio of 10-ml volume per one pot.","After drying the chemical preparations in air, a spore suspension prepared by grinding the spore collected from wheat leave afflicted with Puccinia recondita as the microorganism causing wheat rust was sprayed on the wheat plant, for inoculation.","Then, the plant was left to stand alone in a moist chamber at 22° C. for 15 hours.","Subsequently, the plant was left to stand in a water pool in a green house for 7 days.","Evaluation was carried out by examining the ratio of the diseased area of each leaf to calculate the control ratio by the same method as in Test Example 1.The results are shown in Table 4 (in the table, Compound Nos.","correspond to Compound Nos, in Table-1.)","TABLE 4 Compound No.","Concentration (ppm) Control Ratio (%) 3 500 100 4 500 100 5 500 100 6 500 100 7 500 100 17 500 100 21 500 100 25 500 100 30 500 100 34 500 100 35 500 100 36 500 100 39 500 100 40 500 100 41 500 100 45 500 100 46 500 100 49 500 100 54 500 100 63 500 100 64 500 100 68 500 100 69 500 100 70 500 100 71 500 100 72 500 100 80 500 100 86 500 100 90 500 100 (III) 500 0 Test Example 3 Fungicidal and Bactericidal Effect on Larvae of Spodoptera litura: Compound (III) disclosed in JP-A-64-25763 and various compounds in accordance with the present invention were individually prepared into insecticide preparations according to the formulation of Formulation Example 1.The resulting insecticides of the present invention (hydrates) were diluted with water, where chopped cabbage leave (a diameter of 6 cm) were then immersed for one minute.","After immersion and subsequent drying in air, the cabbage leave were placed in a plastic cup (inner diameter of 7 cm), where five larvae each of 3rd instar Spodoptera litura were left (twice in duplicate per one concentration).","The plastic cup was kept in a thermostat at 25° C. Five days after the larvae were left in the plastic cup, the death and agony of these larvae were examined, to calculate the insecticide activity (%) while an agonizing insect was counted as 0.5 insect death.","The results are shown in Table 5 (in the table, Compound Nos.","correspond to Compound Nos.","in Table-1.)","TABLE 5 Compound No.","Concentration (ppm) Insecticide Activity (%) 3 500 100 5 500 100 6 500 100 7 500 100 15 500 100 37 500 100 39 500 100 41 500 100 42 500 100 47 500 100 49 500 100 52 500 100 55 500 100 70 500 100 78 500 100 86 500 100 90 500 100 100 500 100 107 500 100 (III) 500 20 Test Example 4 Miticidal effect on adult Tetranychus urticae Koch The stem part of a Phaseolus seedling with one fresh first leaf remaining thereon was inserted in a test tube (a volume of 50 ml) containing water, where 15 female adult Tetranychus urticae Koch per one leaf was then inoculated.","One day after the inoculation, the leave deposited with Tetranychus urticae Koch were immersed in and treated with an aqueous dilution of each of miticides (emulsions) individually produced from Compound (III) disclosed in JP-A-64-25763, tebufenpyrade and various compounds of the present invention according to the formulation for Formulation Example 3 (for about 5 seconds) (single one concentration, duplicate) The resulting test tubes were kept in a thermostat at 25° C. Five days after the treatment, the number of the female adult Tetranychus urticae Koch was counted on the Phaseolus leave, to calculate the adult death ratio (%) based on the above results.","The results are shown in Table-6 (Compound Nos.","in the table below correspond to Compound Nos.","in Table-1).","TABLE 6 Compound No.","Concentration (ppm) Adult death ratio (%) 2 500 100 4 500 100 5 500 100 6 500 100 7 500 100 11 500 100 17 500 100 18 500 100 22 500 100 30 500 100 31 500 100 32 500 100 33 500 100 35 500 100 36 500 100 37 500 100 38 500 100 39 500 100 12.5 100 3.1 100 0.8 100 40 500 100 41 500 100 42 500 100 43 500 100 46 500 100 47 500 100 48 500 100 49 500 100 50 500 100 52 500 100 53 500 100 55 500 100 63 500 100 68 500 100 69 500 100 70 500 100 71 500 100 78 500 100 79 500 100 80 500 100 81 500 100 82 500 100 83 500 100 84 500 100 85 500 100 86 500 100 87 500 100 88 500 100 96 500 100 98 500 100 100 500 100 104 500 100 105 500 100 106 500 100 107 500 100 108 500 100 (III) 500 15 tebufenpyrade 12.5 100 3.1 80 0.8 0 While the invention has been described in detail with reference to specific embodiments thereof, it will be apparent to one skilled in the art that various changes and modifications can be made therein without departing from the spirit and scope thereof.","The present application is based on a Japanese patent application filed on Apr.","6, 2001 (Japanese Patent Application 2001-108116), the entire contents of which are incorporated hereinto by reference.","Industrial Applicability The pyrazolecarboxamides of the present invention have such great effects on the control of bacterial strains, hazardous insects and mites that the pyrazolecarboxamides function as great insecticides and miticides for agriculture, forestry and disease control.","Additionally, the pyrazolecarboxamides of the present invention are promising as a control agent of various hazardous biological organisms for livestock industry, fishery, or storage of various products and public hygiene."]],"string":"[\n [\n \"Pyrazolecarboxamides, intermediates therefor and pest control agent comprising the same as active ingredients\",\n \"The present invention provides novel compounds with fungicidal and bactericidal activity as well as insecticidal and miticidal activity, which are useful for controlling hazardous biological organisms and belong to pyrazolecarboxamides represented by the following formula (I): wherein R1 represents C1-C5 alkyl; R2 represents C1-C5 alkyl, C1-C5 haloalkyl, or the like; R3 represents hydrogen or C1-C3 alkyl; R4 represents halogen, C1-C5 alkyl or the like; n is an integer of 0 to 4; R5 represents alkyl, haloalkyl, alkoxy, haloalkoxy or the like; n represents an integer of 1 to 5; and X represents hydrogen, C1-C3 alkyl, C1-C3 alkoxy, halogen, C1-C3 haloalkoxy or nitro.\"\n ],\n [\n \"1.Pyrazolecarboxamide represented by the following formula (I): wherein R1 represents C1-C5 alkyl, R2 represents C1-C5 alkyl, C1-C5 haloalkyl, C1-C5 alkoxy or C1-C5 haloalkoxy, R3 represents hydrogen or C1-C3 alkyl, R4 represents C1-C5 alkyl, C1-C5 haloalkyl, C1-C5 alkoxy, C1-C5 haloalkoxy or halogen, m is an integer of 0 to 4; R5 represents alkyl, haloalkyl, alkoxy, haloalkoxy, alkylthio, haloalkylthio, alkylsulfinyl, haloalkylsulfinyl, alkylsulfonyl, haloalkylsulfonyl, halogen, cyano, formyl, vinyl, alkoxycarbonyl, haloalkoxycarbonyl, acetyl, hydroxycarbonyl, alkylcarbamoyl or a group represented by wherein R7 and R8 each represents hydrogen or C1-C5 alkyl, n represents an integer of 1 to 5, X represents hydrogen, C1-C3 alkyl, C1-C3 alkoxy, halogen, C1-C3 haloalkoxy or nitro, or R2 and X may be taken together to form wherein R9 represents hydrogen or C1-C3 alkyl.\",\n \"2.The pyrazolecarboxamide according to claim 1, wherein R1 in formula (I) is methyl.\",\n \"3.The pyrazolecarboxamide according to claim 1, wherein R2 in formula (I) is ethyl.\",\n \"4.The pyrazolecarboxamide according to claim 1, wherein R5 in formula (I) is C1-C5 haloalkyl or C1-C5 haloalkoxy.\",\n \"5.The pyrazolecarboxamide according to any one of claims 1 to 4 claim 1, wherein R3 in formula (I) is hydrogen.\",\n \"6.The pyrazolecarboxamide according to claim 1, wherein X in formula (I) is hydrogen.\",\n \"7.The pyrazolecarboxamide according to claim 1, wherein, in formula (I), m is an integer of 0 to 2 and n is 1 or 2.8.Phenylbenzylamine represented by the following formula (II): wherein R3 represents hydrogen or C1-C3 alkyl, R4 represents C1-C5 alkyl, C1-C5 haloalkyl, C1-C5 alkoxy, C1-C5 haloalkoxy or halogen, m is an integer of 0 to 4; n represents an integer of 1 to 5; and R6 represents haloalkyl or haloalkoxy, or an acid addition salt thereof.\",\n \"9.A pest control agent comprising the pyrazolecarboxamide according to claim 1 as an active ingredient.\",\n \"10.A fungicidal and bactericidal agent comprising the pyrazolecarboxamide according to claim 1 as an active ingredient.\",\n \"11.An insecticidal and miticidal agent comprising the pyrazolecarboxamide according to claim 1 as an active ingredient thereof.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>In the field of agriculture as well as horticulture and floriculture, various fungicides and bactericides, insecticides and miticides have been developed and used in a practical sense for the purpose of controlling various hazardous insects and plant diseases.\",\n \"However, it cannot be said that agricultural chemicals for common use in the related art have been satisfactory in terms of, for example, effect, spectrum and residual effect or have satisfied the demand to reduce the number and chemical dose to be applied.\",\n \"Additionally, the emergence of hazardous insects and plant diseases with acquired resistance against agricultural chemicals for common use in the related art is now problematic.\",\n \"For the cultivation of vegetables, fruit trees, flowers, teas, wheat, barley, oats, etc., rice plants and the like, various hazardous insects with acquired resistance against diverse types of insecticides of, for example, carbamate-series, pyrethroid-series, benzoyl urea-series, organic chloride-series, and organic phosphorus-series and various pathological microorganisms and bacteria with acquired resistance against various types of fungicides and bactericides of, for example, triazole-series, imidazole-series, pyrimidine-series, benzimidazole-series, dicarboximide-series, and phenylamide-series have emerged in various local regions.\",\n \"Therefore, it has been very tough increasingly year by year to control these various hazardous insects and pathological microorganisms and bacteria causing plant diseases, because of such emergence of these resistant hazardous insects.\",\n \"Thus, it has been consistently desired the development of a novel agricultural chemical of which a lower chemical dose can express a sufficient effect on the control of various pathological microorganisms and bacteria and hazardous insects with acquired resistance against insecticides and fungicides and bactericides for agriculture and horticulture and floriculture in the related art and which shows less adverse effects on environment.\",\n \"The present inventors found that N-benzylpyrazole-5-carboxamide derivatives had insecticidal and miticidal activity.\",\n \"Specifically, the publication of JP-A-64-25763 discloses the following compound with insecticidal and miticidal activity, and the like.\",\n \"Additionally, the publication of JP-A-3-81266 discloses that the following compound with phenoxy and the like have insecticidal and miticidal activity.\",\n \"However, Compound (III) with a structure similar to the compound of the present invention as disclosed in JP-A-64-25763 is so insufficient in view of the insecticidal and miticidal activity that the practical applicability of the compound is still low.\",\n \"Meanwhile, a compound with both fungicidal and bactericidal activity and insecticidal and miticidal activity is very useful for use in agriculture and horticulture and floriculture.\",\n \"Regarding fungicidal and bactericidal activity, it is known that only a compound with specific substituents among compounds described in JP-A-64-25763 has fungicidal and bactericidal activity (JP-A-3-206079).\",\n \"An object of the present invention to provide a new substance with a great effect on the control of various pathological microorganisms and bacteria and with usefulness in the control of insects and mites, particularly a highly safe substance with a great effect on the control of various hazardous insects and pathological bacteria causing plant diseases with acquired resistance against fungicides and bactericides and insecticides in the related art, because such a lower chemical dose thereof can express these effects that problems of residual toxicity and environmental pollution can be reduced.\"\n ],\n [\n \"TECHNICAL FIELD The present invention relates to novel pyrazolecarboxamides and intermediates therefor, and pest control agents comprising the same as an active ingredient.\",\n \"The novel pyrazolecarboxamides in accordance with the present invention are useful as bactericides and fungicides, insecticides and miticides for agriculture, and horticulture and floriculture.\",\n \"BACKGROUND OF THE INVENTION In the field of agriculture as well as horticulture and floriculture, various fungicides and bactericides, insecticides and miticides have been developed and used in a practical sense for the purpose of controlling various hazardous insects and plant diseases.\",\n \"However, it cannot be said that agricultural chemicals for common use in the related art have been satisfactory in terms of, for example, effect, spectrum and residual effect or have satisfied the demand to reduce the number and chemical dose to be applied.\",\n \"Additionally, the emergence of hazardous insects and plant diseases with acquired resistance against agricultural chemicals for common use in the related art is now problematic.\",\n \"For the cultivation of vegetables, fruit trees, flowers, teas, wheat, barley, oats, etc., rice plants and the like, various hazardous insects with acquired resistance against diverse types of insecticides of, for example, carbamate-series, pyrethroid-series, benzoyl urea-series, organic chloride-series, and organic phosphorus-series and various pathological microorganisms and bacteria with acquired resistance against various types of fungicides and bactericides of, for example, triazole-series, imidazole-series, pyrimidine-series, benzimidazole-series, dicarboximide-series, and phenylamide-series have emerged in various local regions.\",\n \"Therefore, it has been very tough increasingly year by year to control these various hazardous insects and pathological microorganisms and bacteria causing plant diseases, because of such emergence of these resistant hazardous insects.\",\n \"Thus, it has been consistently desired the development of a novel agricultural chemical of which a lower chemical dose can express a sufficient effect on the control of various pathological microorganisms and bacteria and hazardous insects with acquired resistance against insecticides and fungicides and bactericides for agriculture and horticulture and floriculture in the related art and which shows less adverse effects on environment.\",\n \"The present inventors found that N-benzylpyrazole-5-carboxamide derivatives had insecticidal and miticidal activity.\",\n \"Specifically, the publication of JP-A-64-25763 discloses the following compound with insecticidal and miticidal activity, and the like.\",\n \"Additionally, the publication of JP-A-3-81266 discloses that the following compound with phenoxy and the like have insecticidal and miticidal activity.\",\n \"However, Compound (III) with a structure similar to the compound of the present invention as disclosed in JP-A-64-25763 is so insufficient in view of the insecticidal and miticidal activity that the practical applicability of the compound is still low.\",\n \"Meanwhile, a compound with both fungicidal and bactericidal activity and insecticidal and miticidal activity is very useful for use in agriculture and horticulture and floriculture.\",\n \"Regarding fungicidal and bactericidal activity, it is known that only a compound with specific substituents among compounds described in JP-A-64-25763 has fungicidal and bactericidal activity (JP-A-3-206079).\",\n \"An object of the present invention to provide a new substance with a great effect on the control of various pathological microorganisms and bacteria and with usefulness in the control of insects and mites, particularly a highly safe substance with a great effect on the control of various hazardous insects and pathological bacteria causing plant diseases with acquired resistance against fungicides and bactericides and insecticides in the related art, because such a lower chemical dose thereof can express these effects that problems of residual toxicity and environmental pollution can be reduced.\",\n \"DISCLOSURE OF THE INVENTION The inventors have investigated the problems described above.\",\n \"Consequently, the inventors have found that novel pyrazolecarboxamides with a biphenyl backbone with substituents and with a specific combination of substituents have fungicidal and bactericidal activity and insecticidal and miticidal activity meeting the features described above.\",\n \"Thus, the present invention has been achieved.\",\n \"The present invention relates to a pyrazolecarboxamide represented by the following formula (I): wherein R1 represents C1-C5 alkyl, R2 represents C1-C5 alkyl, C1-C5 haloalkyl, C1-C5 alkoxy or C1-C5 haloalkoxy, R3 represents hydrogen or C1-C3 alkyl, R4 represents C1-C5 alkyl, C1-C5 haloalkyl, C1-C5 alkoxy, C1-C5 haloalkoxy or halogen, m is an integer of 0 to 4; R5 represents alkyl, haloalkyl, alkoxy, haloalkoxy, alkylthio, haloalkylthio, alkylsulfinyl, haloalkylsulfinyl, alkylsulfonyl, haloalkylsulfonyl, halogen, cyano, formyl, vinyl, alkoxycarbonyl, baloalkoxycarbonyl, acetyl, hydroxycarbonyl, alkylcarbamoyl or a group represented by wherein R7 and R8 each represents hydrogen or C1-C5 alkyl, n represents an integer of 1 to 5, X represents hydrogen, C1-C3 alkyl, C1-C3 alkoxy, halogen, C1-C3 haloalkoxy or nitro, or R2 and X may be taken together to form wherein R9 represents hydrogen or C3-C3 alkyl, a pest control agents comprising the pyrazolecarboxamide as an active ingredient, a phenylbenzylamine as an intermediate for the pyrazolecarboxamide, which is represented by the following formula (II): wherein R3, R4, m and n have the same meanings as defined in formula (I); R represents haloalkyl or haloalkoxy, and an acid addition salt thereof.\",\n \"BEST MODE FOR CARRYING OUT THE INVENTION The present invention is now described in detail below.\",\n \"Pyrazolecarboxamides: In the pyrazolecarboxamides represented by formula (I) in accordance with the present invention, the substituent R1 represents a linear or branched C1-C5 alkyl such as methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, 2-methylbutyl, 3-methylbutyl, isopentyl, 2-ethylpropyl, neopentyl, 2,2-dimethylpropyl, or 2-methylisobutyl.\",\n \"Among these, methyl is preferable as the substituent R1.The substituent R2 represents linear, branched or cyclic C1-C5 alkyl such as methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, 2-methylbutyl, 3-methylbutyl, isopentyl, 2-ethylpropyl, neopentyl, 2,2-dimethylpropyl, 2-methylisobutyl, or cyclopropyl; linear or branched C1-C5 haloalkyl such as fluoronethyl, chloromethyl, bromomethyl, difluoromethyl, difluoroethyl, trifluoromethyl, trichloromethyl, tribromomethyl, 2-fluoroethyl, 2,2,2-trifluoroethyl, 1-(trifluoromethyl)ethyl, 1,1,2,3,3,3-hexafluoropropyl, 2,2,3,3,3-pentafluoropropyl, 2,2,3,3-tetratluoropropyl, 2,2,3,3,4,4,4-heptafluorobutyl, or 2,2,3,3,4,4,5,5,5-nonafluoropentyl; linear or branched C1-C5 alkoxy such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, isobutoxy, tert-butoxy, n-pentoxy, 2-methylbutoxy, 3-methylbutoxy, isopentoxy, 2-ethylpropoxy, neopentoxy, 2,2-dimethylpropoxy, or 2-methylisobutoxy; or linear or branched C1-C5 haloalkoxy such as difluoromethoxy, trifluoromethoxy, fluoromethoxy, trichloromethoxy, tribromomethoxy, 2-fluoroethoxy, 2,2,2-trifluoroethoxy, 2,2,2-trichloroethoxy, 1-(trifluoromethyl)ethoxy, 1,1,2,3,3,3-hexafluoropropoxy, 2,2,3,3,3-pentafluoropropoxy, or 2,2,3,3-tetrafluoropropoxy.\",\n \"Among these, linear C1-C3 alkyl is preferable as the substituent R2 Ethyl is particularly preferable.\",\n \"The substituent R3 represents hydrogen; or linear or branched C1-C3 alkyl such as methyl, ethyl, n-propyl, or isopropyl.\",\n \"Among these, hydrogen is preferable as the substituent R3.The substituent R4 represents linear or branched C1-C5 alkyl such as methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, 2-methylbutyl, 3-methylbutyl, isopentyl, 2-ethylpropyl, neopentyl, 2,2-dimethylpropyl, or 2-methylisobutyl; linear or branched C1-C5 haloalkyl such as fluoromethyl, chloromethyl, bromomethyl, difluoromethyl, difluoroethyl, trifluoromethyl, trichloromethyl, tribromomethyl, 2-fluoroethyl, 2,2,2-trifluoroethyl, 1-(trifluoromethyl)ethyl, 1,1,2,3,3,3-hexafluoropropyl, 2,2,3,3,3-pentafluoropropyl, 2,2,3,3-tetrafluoropropyl, 2,2,3,3,4,4,4-heptafluorobutyl, or 2,2,3,3,4,4,5,5,5-nonafluoropentyl; linear or branched C1-C5 alkoxy such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, isobutoxy, text-butoxy, n-pentoxy, 2-methylbutoxy, 3-methylbutoxy, isopentoxy, 2-ethylpropoxy, neopentoxy, 2,2-dimethylpropoxy, or 2-methylisobutoxy; linear or branched C1-C5 haloalkoxy such as difluoromethoxy, trifluoromethoxy, fluoromethoxy, trichloromethoxy, tribromomethoxy, 2-fluoroethoxy, 2,2,2-trifluoroethoxy, 2,2,2-trichloroethoxy, 1-(trifluoromethyl)ethoxy, 1,1,2,3,3,3-hexafluoropropoxy, 2,2,3,3,3-pentafluoropropoxy, or 2,2,3,3-tetrafluoropropoxy; or halogen such as fluorine, chlorine, bromine or iodine.\",\n \"Among these, halogen is preferable as the substituent R4.The substituent R5 represents linear or branched alkyl such as methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, 2-methylbutyl, 3-methylbutyl, isopentyl, 2-ethylpropyl, neopentyl, 2,2-dimethylpropyl, 2-methylisobutyl, n-hexyl, n-heptyl or n-octyl; linear or branched haloalkyls such as fluoromethyl, chloromethyl, bromomethyl, difluoromethyl, difluoroethyl, trifluoromethyl, trichloromethyl, tribromomethyl, 2-fluoroethyl, 2,2,2-trifluoroethyl, 1-(trifluoromethyl)ethyl, 1,1,2,3,3,3-hexafluoropropyl, 2,2,3,3,3-pentafluoropropyl, 2,2,3,3-tetrafluoropropyl, 2,2,3,3,4,4,4-heptafluoropropyl, or 2,2,3,3,4,4,5,5,5-nonafluoropentyl; linear or branched alkoxys such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, isobutoxy, tert-butoxy, n-pentoxy, 2-methylbutoxy, 3-methylbutoxy, isopentoxy, 2-ethylpropoxy, neopentoxy, 2,2-dimethylpropoxy, 2-methylisobutoxy, n-hexyloxy, n-heptyloxy, or n-octyloxy; linear or branched haloalkoxy such as difluoromethoxy, trifluoromethoxy, fluoromethoxy, trichloromethoxy, tribromoinethoxy, 2-fluoroethoxy, 2,2,2-trifluoroethoxy, 2,2,2-trichloroethoxy, 1-(trifluoromethyl)ethoxy, 1,1,2,3,3,3-hexafluoropropoxy, 2,2,3,3,3-pentafluoropropoxy, 2,2,3,3-tetrafluoropropoxy, or 2,2,3,3,4,4,4-heptafluorobutoxy; linear or branched alkylthio such as methylthio, ethylthio, propylthio, isopropylthio, butylthio, sec-butylthio, isobutylthio, tert-butylthio, pentylthio, 2-methylbutylthio, 3-methylbutylthio, isopentylthio, 2-ethylpropylthio, neopentylthio, 2,2-direthylpropylthio, or 2-methylisobutylthio; haloalkylthio such as trifluoronethylthio, perfluoroethylthio, perfluoropropylthio, or 2,2,2-trifluoroethylthio; linear or branched alkylsulfinyl such as methylsulfinyl, ethylsulfinyl, propylsulfinyl, isopropylsulfinyl, butylsulfinyl, sec-butylsulfinyl, isobutylsulfinyl, tert-butylsulfinyl, pentylsulfinyl, 2-methylbutylsulfinyl, 3-methylbutylsulfinyl, isopentylsulfinyl, 2-ethylpropylsulfinyl, neopentylsulfinyl, 2,2-dimethylpropylsulfinyl or 2-methylisobutylsulfinyl; haloalkylsulfinyl such as trifluoromethylsulfinyl or 2,2,2-trifluoroethylsulfinyl; linear or branched alkylsulfonyl such as methylsulfonyl, ethylsulfonyl, propylsulfonyl, isopropylsulfonyl, butylsulfonyl, sec-butylsulfonyl, isobutylsulfonyl, text-butylsulfonyl, pentylsul fonyl, 2-methylbutylsulfonyl, 3-methylbutylsulfonyl, isopentylsulfonyl, 2-ethylpropylsulfonyl, neopentylsulfonyl, 2,2-dimethylpropylsulfonyl, or 2-methylisobutylsulfonyl; haloalkylsulfonyl such as trifluorosulfonyl or 2,2,2-trifluoroethylsulfonyl; halogen such as fluorine, chlorine, bromine or iodine; cyano; formyl; vinyl; linear or branched alkoxycarbonyl such as methoxycarbonyl, ethoxycarbonyl, n-propoxycarbonyl, isopropoxycarbonyl or butoxycarbonyl; haloalkoxycarbonyl such as 2,2,2-trifluoromethoxycarbonyl or 2,2,3,3,3-pentafluorpropoxycarbonyl; acetyl; hydroxycarbonyl; alkylcarbamoyl such as methylcarbamoyl, ethylcarbamoyl or propylcarbamoyl; or a group represented by Herein, the above R7 and R8 each represents hydrogen; or linear or branched C1-C5 alkyl such as methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, or pentyl.\",\n \"In the above R5, the alkyl and alkoxy have preferably C1-C8; the haloalkyl, haloalkoxy, alkylthio, haloalkylthio, alkylsulfinyl, haloalkylsulfinyl, alkylsulfonyl and haloalkylsulfonyl have preferably C1-C5.The alkoxycarbonyl, haloalkoxycarbonyls, alkylcarbamoyls and group represented by have preferably C2-C5, more preferably C2-C5.Among these, the substituent R5 is preferably haloalkyl or haloalkoxy, more preferably C1-C5 haloalkyl or C1-C5 haloalkoxy, particularly preferably trifluoromethoxy or difluoromethoxy.\",\n \"Additionally, the substituent is preferably positioned at the 4-position.\",\n \"The substituent X represents hydrogen; linear or branched C1-C3 alkyl such as methyl, ethyl, n-propyl, and isopropyl; linear or branched C1-C3 alkoxy such as methoxy, ethoxy, n-propoxy, or isopropoxy; halogen such as fluorine, chlorine, bromine, or iodine; C1-C3 haloalkoxy such as difluoromethoxy, 2,2,2-trifluoroethoxy, or perfluoropropoxy; or nitro.\",\n \"Among these, the substituent X is preferably hydrogen.\",\n \"Furthermore, R9 in the group formed by R2 and X together represents hydrogen; or C1-C3 alkyl such as methyl, ethyl, n-propyl or isopropyl.\",\n \"m is an integer of 0 to 4, preferably 0 to 2; and n is an integer of 1 to 5, preferably 1 or 2.Phenylbenzylamines: Furthermore, the phenylbenzylamines represented by formula (II) and acid addition salts thereof in accordance with the present invention are novel intermediates for producing the pyrazolecarboxamides.\",\n \"Herein, the substituents R3 and R4, m and n have the same meanings as defined in formula (I).\",\n \"The substituent R6 represents haloalkyl such as fluoromethyl, chloromethyl, bromomethyl, difluoromethyl, difluoroethyl, trifluoromethyl, trichloromethyl, tribromomethyl, 2-fluoroethyl, 2,2,2-trifluoroethyl, 1-(trifluoromethyl)ethyl, 1,1,2,3,3,3-hexafluoropropyl, 2,2,3,3,3-pentafluoropropyl, 2,2,3,3-tetrafluoropropyl, 2,2,3,3,4,4,4-heptafluorobutyl, or 2,2,3,3,4,9,5,5,5-nonafluoropentyl; or haloalkoxys such as difluoromethoxy, trifluoromethoxy, fluoromethoxy, trichloromethoxy, tribromomethoxy, 2-fluoroethoxy, 2,2,2-trifluoroethoxy, 2,2,2-trichloroethoxy, 1-(trifluoroimethyl)ethoxy, 1,1,2,3,3,3-hexafluoropropoxy, 2,2,3,3,3-pentafluoropropxy, 2,2,3,3-tetrafluoropropoxy, or 2,2,3,3,4,4,4-heptafluorobutoxy.\",\n \"The haloalkyl and haloalkoxy have preferably C1-C5.The acid to be added to the phenylbenzylamines represented by formula (II) in accordance with the present invention includes carboxylic acid such as acetic acid, propionic acid, butyric acid, oxalic acid, adipic acid, dodecanedioic acid, lauric acid, stearic acid, trifluoroacetic acid, fumaric acid, maleic acid, benzoic acid, or phthalic acid; sulfonate such as methanesulfonic acid, 1,3-propanedisulfonic acid, p-toluenesulfonic acid, or dodecylbenzenesulfonic acid; and inorganic acid such as hydrochloric acid, sulfuric acid, nitric acid, or carbonic acid.\",\n \"Production Process: The compound represented by formula (I) in accordance with the present invention is a novel compound and can be produced, for example, by the following reaction scheme.\",\n \"(In the formula, R1, R2, R3, R4, R5, X, m and n have the same meanings as described in formula (I).)\",\n \"In other words, pyrazolecarboxylic acid chloride (IV) and phenylbenzylamine (V) are allowed to react in the presence or absence of a base, preferably using a solvent at −10 to 50° C., preferably 0 to 25° C. As the solvent, any solvent can be used, so long as it does not have direct influence on the reaction.\",\n \"The solvent includes aromatic hydrocarbons such as benzene, toluene and xylene; ketones such as acetone, methyl ethyl ketone, and methyl isobutyl ketone; halogenated hydrocarbons such as chloroform and dichloromethane; water; esters such as methyl acetate and ethyl acetate; and polar solvents such as tetrahydrofuran, acetonitrile, dioxane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, and pyridine.\",\n \"The base includes alkali metal hydrides such as sodium hydride; alkali metal hydroxides such as sodium hydroxide and potassium hydroxide; alkali metal carbonates such as sodium carbonate and potassium carbonate; inorganic salts; and amines such as pyridine and triethylamine.\",\n \"In order to isolate the objective compound of formula (I) after the reaction, the following procedures are carried out in case of using a solvent soluble in water: distilling off the solvent under reduced pressure, adding water to the resulting residue, extracting the residue in aromatic hydrocarbons insoluble in water such as benzene, toluene and xylene, halogenated hydrocarbons such as chloroform and dichloromethane, and esters such as ethyl acetate, rinsing the extract with a saturated aqueous sodium chloride solution, drying the extract over desiccants such as anhydrous sodium sulfate or anhydrous magnesium sulfate and then distilling off the solvents under reduced pressure.\",\n \"When a solvent insoluble in water is used, water is added to the reaction mixture, followed by separation; the resulting organic layer is rinsed with a saturated aqueous sodium chloride solution and dried over desiccants such as anhydrous sodium sulfate or magnesium sulfate, from which the solvent is distilled off under reduced pressure.\",\n \"The residue recovered after distillation of the solvent is purified by recrystallization, suspending and rinsing, and column chromatography, so that the objective compound represented by formula (I) can be recovered.\",\n \"The compound represented by formula (IV) as one raw material can be synthetically prepared, for example, by the method described in JP-A-64-25763.The compound represented by formula (V) as one raw material can be synthetically prepared, for example, by the following method.\",\n \"(1) Reduction of Phenylbenzonitriles (In the formula, R4, R5, in and n have the same meanings as described in formula (I).)\",\n \"Phenylbenzonitrile (VI) and hydrogen are allowed to react in the presence of a metal catalyst and aqueous ammonia, preferably using an organic solvent, at 0 to 100° C., preferably 25 to 50° C. As the organic solvent, any organic solvent can be used, so long as it does not have directly influence on the reaction.\",\n \"The organic solvent includes alcohols such as methanol, ethanol and isopropanol; aromatic hydrocarbons such as benzene, toluene and xylene; ketones such as acetone, methyl ethyl ketone, and methyl isobutyl ketone; halogenated hydrocarbons such as chloroform and dichloromethane; water; esters such as methyl acetate and ethyl acetate; or polar solvents such as tetrahydrofuran, acetonitrile, dioxane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, and pyridine.\",\n \"The metal catalyst includes Raney-nickel, palladium carbon, and platinum oxide.\",\n \"In order to isolate the objective compound of formula (Va) after the reaction, the metal catalyst is filtered off from the reaction mixture, and the solvent is distilled off from the resulting filtrate under reduced pressure.\",\n \"The resulting product may be sufficiently pure as it is, but the product can be purified by means of evaporation and column chromatography, to recover a pure product.\",\n \"Phenylbenzonitrile (VI) can be synthetically prepared by reaction of the corresponding haloallene with phenylboronic acid in the presence of a zero-valent palladium catalyst and a base according to known methods, for example, the method described in Synthetic Communications, Vol.\",\n \"11, page 513 (1981).\",\n \"(2) Reduction of Oximes Oxime (VII) and hydrogen are allowed to react in the presence of a metal catalyst and aqueous ammonia, preferably using an organic solvent, at 0 to 100° C., preferably 25 to 50° C. As the organic solvent, any organic solvent can be used, so long as it has not direct influence on the reaction.\",\n \"The organic solvent includes alcohols such as methanol, ethanol and isopropanol; aromatic hydrocarbons such as benzene, toluene and xylene; ketones such as acetone, methyl ethyl ketone, and methyl isobutyl ketone; halogenated hydrocarbons such as chloroform and dichlioromethane; water; esters such as methyl acetate and ethyl acetate; and polar solvents such as tetrahydrofuran, acetonitrile, dioxane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, and pyridine.\",\n \"The metal catalyst includes Raney-nickel, palladium carbon, and platinum oxide.\",\n \"In order to isolate the objective compound of formula (Vb) after the reaction, the metal catalyst is filtered off from the reaction mixture, and the solvent is distilled off from the resulting filtrate under reduced pressure.\",\n \"Oxime (VII) can be synthetically prepared by reaction of the corresponding acetophenone with hydroxyamine in the presence of potassium carbonate according to known methods.\",\n \"Phenylbenzylamine-acid addition salts (Vc) can be synthesized by the following method.\",\n \"Phenylbenzylamine (V) and acid (Y) are allowed to react, preferably using a solvent, at −5 to 50° C., preferably 0 to 25° C. As the acid, organic acid and inorganic acid can be used.\",\n \"The organic acid includes carboxylic acids such as acetic acid, propionic acid, butyric acid, oxalic acid, adipic acid, dodecanedioic acid, lauric acid, stearic acid, trifluoroacetic acid, fumaric acid, maleic acid, benzoic acid, and phthalic acid; and sulfonates such as methanesulfonic acid, 1,3-propanedisulfonic acid, p-toluenesulfonic acid, and dodecylbenzenesulfonic acid.\",\n \"The inorganic acid includes hydrochloric acid, sulfuric acid, nitric acid and carbonic acid.\",\n \"As the solvent, any solvent can be used, so long as it does not have direct influence on the reaction.\",\n \"The organic solvent includes aromatic hydrocarbons such as benzene, toluene and xylene; ketones such as acetone, methyl ethyl ketone, and methyl isobutyl ketone; halogenated hydrocarbons such as chloroform and dichloromethane; water; esters such as methyl acetate and ethyl acetate; and polar solvents such as tetrahydrofuran, acetonitrile, dioxane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, and pyridine.\",\n \"After the reaction, the solvent is distilled off under reduced pressure.\",\n \"The resulting crystals are filtered and recovered, to synthetically prepare inorganic acid salts of benzylamine and organic acid salts thereof.\",\n \"The crystals are satisfactory as they are.\",\n \"For further purification, however, the crystals are rinsed with the organic solvents described above, to synthetically prepare benzylamine salts at high purity.\",\n \"Pest Control Agents: The compound represented by formula (I) in accordance with the present invention has high fungicidal and bactericidal effects on pathological microorganisms and bacteria against plants such as Pyricularia oxyzae, pathological microorganisms and bacteria causing powdery mildew, pathological microorganisms and bacteria causing rust, and pathological microorganisms and bacteria causing downy mildew so that the compound is useful as an active ingredient for fungicides and bactericides for agriculture and horticulture and floriculture.\",\n \"Since the compound of the present invention has high activity on the control of the eggs and larvae of insects of the order Hemiptera including leafhoppers such as Sogatella furcifera Horvath, Nilaparvata lugens (Stal), and Laodelphax stratella (Fallen); Cicadomorpha-2 such as Bothrogonia ferruginea (Fabricius) and Cicadella viridis (Linnaeus); and Aphididae such as Myzus persicae; the order Lepidoptera including Spodoptera litura Fabricius, Chilo suppresalis, Cnaphalocrosis medinalis Guenee and Plutella xylostella (Linnaeus); the order Coleoptera including Callosobruchus chinensis; the order Diptera including Musca domestica Linnaeus, Aedes aegypti and Culex pipiens pallens; the order Orthoptera; and the order Acarina including Tetranychus urticae Koch, Tetranychus cinnabarinus (Boisduval) and Panonychus citri (Mcgregor), the compound of the present invention is useful as an active ingredient for insecticides and miticides for agriculture and horticulture and floriculture.\",\n \"It is needless to say that the pathological microorganisms and bacteria against plants, insects, and mites as subjects to be controlled with the compound of the present invention are not limited to those described above.\",\n \"When the compound represented by formula (I) in accordance with the present invention is used for fungicides and bactericides, insecticides and/or miticides for agriculture and horticulture and floriculture, the compound may satisfactorily be used singly.\",\n \"However, preferably, the compound may be used in composition forms produced using agricultural auxiliary agents for common use in the art, such as fungicides and bactericides, insecticides and/or miticide's for agriculture and horticulture and floriculture.\",\n \"The forms are preferably, for example, emulsions, hydrates, powders, flowable agents, fine granules, granules, tablets, oils, sprays, and fumes.\",\n \"However, the forms are not limited thereto.\",\n \"One or two or more forms of the compound may also be blended as active ingredients.\",\n \"The agricultural auxiliary agents for use in producing the fungicides and bactericides, insecticides and/or miticides for agriculture and horticulture and floriculture are used, for example, for the improvement of the effects of fungicides and bactericides, insecticides and/or miticides for agriculture and horticulture and floriculture and for the improvement of stability and dispersibility.\",\n \"For example, carriers (diluents), spreading agents, emulsifiers, wet spreading agents, dispersants and disintegrators can be used.\",\n \"The liquid carriers include water, aromatic hydrocarbons such as toluene and xylene, alcohols such as methanol, butanol, and glycol, ketones such as acetone, amides such as dimethylformamide, sulfoxides such as dimethyl sulfoxide, methylnaphthalene, cyclohexane, animal and vegetable oils, and fatty acid.\",\n \"Additionally, the solid carriers include clay, kaolin, talc, diatomaceous earth, silica, calcium carbonate, montmollionite, bentonite, feldspar, quartz, alumina, sawdust, nitrocellulose, starch and gum arabic.\",\n \"As the emulsifiers and the dispersants, general surfactants can be used and include anionic surfactants, cationic surfactants, nonionic surfactants and ampholytic surfactants, such as higher alcohol sodium sulfate, stearyltrimethylammonium chloride, polyoxyethylene alkylphenyl ether, and lauryl betaine.\",\n \"Additionally, the following agents can also be used: spreading agents such as polyoxyethylene nonyl phenyl ether and polyoxyethylene lauryl phenyl ether; wet spreading agents such as dialkylsulfosuccinate; attaching agents such as carboxymethyl cellulose and polyvinyl alcohol; and disintegrators such as sodium lignin sulfonate and sodium lauryl sulfate.\",\n \"The content of the active ingredient in the fungicides and bactericides, insecticides and/or miticides for agriculture and horticulture and floriculture in accordance with the present invention is selected within a range of 0.1 to 99.5% and is appropriately determined on the basis of various conditions for preparations and application methods.\",\n \"Preferably, for example, powders are produced so as to contain the active ingredient at about 00.5 to 20% by weight, preferably about 10% by weight; hydrates are produced so as to contain the active ingredient at about 1 to 90% by weight, preferably 10 to 80% by weight; emulsions are produced so as to contain the active ingredient at about 1 to 90% by weight, preferably about 10 to 40% by weight.\",\n \"In the case of the emulsions, for example, solvents, surfactants and the like are mixed with the compound as the active ingredient to prepare a stock emulsion.\",\n \"The stock emulsion is diluted with water to a given concentration when used, and is applied.\",\n \"The resulting solution can be applied.\",\n \"In the case of the hydrates, the compound as the active ingredient, a solid carrier, a surfactant and the like are mixed together to prepare a stock solution.\",\n \"The stock solution is diluted with water when used, and is applied.\",\n \"In the case of the powders, the compound as the active ingredient, a solid carrier and the like are mixed together.\",\n \"The resulting mixture can be used as it is.\",\n \"In the case of the granules, the compound as the active ingredient, a solid carrier, a surfactant and the like are mixed together and granulated, to prepare granules, which can be used as it is.\",\n \"It is needless to say that the methods for preparing the preparations are not limited to those described above.\",\n \"One skilled in the art can select an appropriate method, depending on the type of the active ingredient, the purpose of application and the like.\",\n \"In addition to the compound as the active ingredient, the fungicides and bactericides, the insecticides and/or miticides for agriculture and horticulture and floriculture in accordance with the present invention may contain any appropriate active ingredients such as other fungicides and bactericides, insecticides, miticides, herbicides, insect growth controlling agents, fertilizers, and soil modifiers.\",\n \"The method for applying the fungicides and bactericides, insecticides and/or miticides for agriculture and horticulture and floriculture in accordance with the present invention is not particularly limited, and spraying on stem and leaf, application on water surface, soil treatment, and seed treatment can be carried out.\",\n \"In the case of spraying on stem and leaf, for example, a solution within a concentration range of 5 to 1,000 ppm, preferably 10 to 500 ppm can be used at a volume of about 100 to 200 liters per 10 Ares.\",\n \"For application on water surface, generally, granules at 5-15% content of the active ingredient are applied at a ratio of 1 to 10 kg per 10 ares.\",\n \"In the case of soil treatment, a solution within a concentration range of 5 to 1,000 ppm is applied at a ratio of about 1 to 10 liters per 2, In the case of seed treatment, a solution within a concentration range of 10 to 1,000 ppm is applied at a ratio of about 10 to 100 ml per kg of seeds.\",\n \"The present invention is now described in the following examples in more detail.\",\n \"However, the scope of the present invention is not limited to the following examples.\",\n \"EXAMPLES The present invention is now described below in detail in the following examples, Formulation Examples and test examples.\",\n \"However, the present invention is not limited to the examples, unless the examples are departed from the scope of the present invention.\",\n \"Example 1 N-[4-(4-Difluoromethoxyphenyl)benzyl]-1-methyl-3-ethylpyrazole-5-carboxamide: A mixture of 4-(4-difluoromethoxyphenyl)benzylamine (13.8 g), triethylamine (10 ml) and dichloromethane (100 ml) was cooled to 0° C. to 4° C., followed by addition of a solution (20 ml) of 1-methyl-3-ethylpyrazole-5-carboxylic acid chloride (12.4 g) in dichloromethane, for agitation for 30 minutes.\",\n \"The reaction mixture was poured into water (200 ml), for extraction in dichloromethane.\",\n \"The organic layer was rinsed with a saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfate.\",\n \"The solvent was distilled off under reduced pressure.\",\n \"The resulting residue was purified by silica gel chromatography, to obtain 17.0 g of Compound No.\",\n \"30 described in Table-1.The melting point was 107 to 108° C. Example 2 According to the method in Example 1, the compounds described in Table-1 were prepared synthetically.\",\n \"TABLE 1 Compound No.\",\n \"R1 R2 X R3 m.p.\",\n \"(° C.) or nD/(° C.) 1 CH3 CH3 CH3 H 162-163 2 CH3 CH3 C2H5O H 1.5464/23.0 3 CH3 CH3 H H 128-130 4 CH3 CH3 H H 98-99 5 CH3 CH3 H H 134-135 6 CH3 CH3 H H 127-128 7 CH3 CH3 H H 122-124 8 CH3 C2H5 H H 116-118 9 CH3 C2H5 H H 119-121 10 CH3 C2H5 H H 106-107 11 CH3 C2H5 H H 104-108 12 CH3 C2H5 H H 105-107 13 CH3 C2H5 H H 110-111 14 CH3 C2H5 H H 90-91 15 CH3 C2H5 H H 95-97 16 CH3 C2H5 H H 1.5671/23.0 17 CH3 C2H5 H H 143-145 18 CH3 C2H5 Cl H 124-125 19 CH3 C2H5 H H 132-133 20 CH3 C2H5 Cl H 142-143 21 CH3 C2H5 H H 143-144 22 CH3 C2H5 Cl H 155-156 23 CH3 C2H5 H H 151-153 24 CH3 C2H5 H H 124-126 25 CH3 C2H5 H H 122-123 26 CH3 C2H5 H H 126-127 27 CH3 C2H5 H H 122-123 28 CH3 C2H5 H H 96-97 29 CH3 C2H5 H H 91-93 30 CH3 C2H5 H H 107-108 31 CH3 C2H5 F H 124-125 32 CH3 C2H5 H H 1.5623/25.0 33 CH3 C2H5 H H 109-110 34 CH3 C2H5 H CH3 81-82 35 CH3 C2H5 F H 105-106 36 CH3 C2H5 Cl H 93-95 37 CH3 C2H5 H H 114-116 38 CH3 C2H5 Cl H 117-118 39 CH3 C2H5 H H 1.5523/23.0 40 CH3 C2H5 Cl H 96-97 41 CH3 C2H5 H H 95-96 42 CH3 C2H5 Cl H 106-107 43 CH3 C2H5 H H 95-96 44 CH3 C2H5 H CH3 122-123 45 CH3 C2H5 H H 99-100 46 CH3 C2H5 Cl H 108-109 47 CH3 C2H5 H H 113-114 48 CH3 C2H5 Cl H 106-107 49 CH3 C2H5 H H 118-119 50 CH3 C2H5 H H 163-164 51 CH3 C2H5 Cl H 136-137 52 CH3 C2H5 H H 119-121 53 CH3 C3H5 H H 141-143 54 CH3 C2H5 H H 154-156 55 CH3 C2H5 H CH3 147-150 56 CH3 C2H5 H H 1.5865/25.0 57 CH3 i-C3H7 Br H 142-143 58 CH3 CF3 H H 153-154 59 C2H5 C2H5 H H 127-128 60 i-C3H7 C2H5 H H 1.5520/25.0 61 CH3 C2H5 H H 1.5530/23.0 62 CH3 C3H5 H H 1.5671/25.0 63 CH3 C2H5 H H 81-83 64 CH3 C2H5 F H 101-104 65 CH3 C2H5 H H 87-89 66 CH3 C2H5 H H 1.5476/23.0 67 CH3 C2H5 H H 123-124 68 CH3 CH3 H H 111-112 69 CH3 i-C3H7 H H 120-122 70 CH3 H H 99-100 71 CH3 CH3 H H 88-89 72 CH3 C2H5 Cl H 106-107 73 CH3 C2H5 H H 117-118 74 CH3 C2H5 H H 155-157 75 CH3 C2H5 H H 130-131 76 CH3 C2H5 H H 167-168 77 CH3 C2H5 H H 245-247 78 CH3 C2H5 H H 140-141 79 CH3 C2H5 Cl H 154-156 80 CH3 C2H5 H H 102-104 81 CH3 CH3 H H 128-129 82 CH3 C2H5 Cl H 122-123 83 CH3 C2H5 H H 155-160 84 CH3 n-C3H7 H H 131-135 85 CH3 t-C4H9 H H 123-125 86 CH3 CH3 CH3O H 75-77 87 CH3 CH3 n-C3H7O H 51-54 88 CH3 CH3 CHF2O H 136-138 89 CH3 C2H5 H H 209-211 90 CH3 C2H5 H H 139-141 91 CH3 C2H5 H H 242-245 92 CH3 C2H5 H H 162-164 93 CH3 C2H5 H H 152-154 94 CH3 C2H5 H H 146-147 95 CH3 C2H5 H H 131-136 96 CH3 CH3O H H 124-125 97 CH3 C2H5O H H 104-105 98 CH3 i-C3H7O H H 1.5480/25.0 99 CH3 CHF2O H H 137-139 100 CH3 CH3 Cl H 117-118 101 CH3 n-C3H7O H H 81-83 102 CH3 CH3 NO2 H 198-199 103 CH3 H H H 123-125 104 CH3 H 155-156 105 CH3 H 157-159 106 CH3 H 194-195 107 CH3 H 190-192 108 CH3 CH3 H H 150-151 Example 3 Synthesis of 4-(4-difluoromethoxyphenyl)benzylamine: A mixture of 4-cyano-4′-difluoromethoxybiphenyl (15.0 g), Raney-nickel R-100 (15 g) and aqueous 28% ammonia (15.0 ml) was shaken in hydrogen atmosphere at 50° C. for 4 hours.\",\n \"After the reaction mixture was cooled to ambient temperature, Raney-nickel was filtered off.\",\n \"The solvent was distilled off under reduced pressure, to obtain 13.8 g of Compound No.\",\n \"113 described in Table-2.The melting point was 109 to 11° C. Example 4 Synthesis of 4-(4-trifluoromethoxy)-α-methylbenzylamine: A mixture of 4-(4-trifluoromethoxyphenyl)acetophenone oxime (8.8 g), Raney-nickel R-100 (10 g) and aqueous 28% ammonia (10.0 ml) was shaken in hydrogen atmosphere at 50° C. for 3 hours.\",\n \"After the reaction mixture was cooled to ambient temperature, Raney-nickel was filtered off.\",\n \"The solvent was distilled off under reduced pressure, to obtain 7.61 g of Compound No.\",\n \"117 described in Table-2.The melting point was 70 to 74° C. Example 5 Synthesis of 4-(4-trifluoromethoxyphenyl)benzylamine Hydrochloride: To a solution (20 ml) of 4-(4-trifluoromethoxyphenyl)benzylamine (5 g) in methanol, 12N hydrochloric acid (3.0 ml) was added, followed by agitation at ambient temperature for 2 hours, The solvent was distilled off under reduced pressure.\",\n \"The resulting residue was rinsed with methanol, to obtain 4.8 g of Compound No.\",\n \"116 described in Table-2.The melting point was 252 to 254° C. Example 6 According to the methods described in Examples 3 to 5, the compounds described in Table-2 were synthetically prepared.\",\n \"TABLE 2 Compound No.\",\n \"R3 Y m.p.\",\n \"(° C.) or nD/(° C.) 109 H — 1.5459/23.0 110 H — 1.5530/23.0 111 H — 143-145 112 H — 109-110 113 H — 109-110 114 H HCl 220-225 115 H — 80-82 116 H HCl 252-254 117 CH3 — 70-74 118 H — 1.5430/23.0 119 H — 1.5323/23.0 120 H — 1.4911/23.0 121 H HCl 176-180 122 H — 1.5488/23.0 123 H — 1.5030/23.0 124 H — 1.5350/23.0 125 H HCl 302-303 126 H — 1.5540/23.0 127 H — 87-89 128 H — 1.5140/25.0 129 H — 1.5531/25.0 130 H — 1.5380/23.0 131 H — 1.5273/25.0 132 H — 1.5222/25.0 133 H — 1.5301/23.0 *Compound No.130 b.p.\",\n \"= 134-135° C./1 mmHg Hereinbelow, Formulation Examples containing the compound of the present invention as the active ingredient are described, which are insecticides and miticides for agriculture and horticulture and floriculture.\",\n \"However, the mode for carrying out the present invention is not limited to those described below.\",\n \"Formulation Example 1 Hydrates: Twenty parts by weight of the compound of the present invention, 20 parts by weight of Carplex #80 (trade name; white carbon; manufactured by Shionogi & Co., Ltd.), 52 parts by weight of ST kaolin clay (trade name; kaolinite; manufactured by Tuchiya Kaolin Co., Ltd.), 5 parts by weight of Solpol 9047K (trade name; anionic surfactant; manufactured by Toho Chemical, Co., Ltd.) and 3 parts by weight of Lunox P56L (trade name; anionic surfactant; Toho Chemical Co., Ltd.) were blended together, uniformly mixed together and ground, to obtain hydrates containing the active ingredient at a content of 20% by weight.\",\n \"Formulation Example 2 Powders; Two parts by weight of the compound of the present invention, 93 parts by weight of clay (manufactured by Nippon Talc Co., Ltd.), and 5 parts by weight of Carplex #80 (trade name; white carbon; manufactured by Shionogi Pharmaceutical Co., Ltd.) were uniformly mixed together and ground, to obtain powders containing the active ingredient at a content of 2% by weight.\",\n \"Formulation Example 3 Emulsions: Twenty parts by weight of the compound of the present invention were dissolved in a mixture solvent of 35 parts by weight of xylene and 30 parts by weight of dimethylformamide, followed by addition of 15 parts by weight of Solpol 3005X (trade name; a mixture of a nonionic surfactant and an anionic surfactant; Toho Chemical Co., Ltd.), to obtain emulsions containing the active ingredient at a content of 20% by weight.\",\n \"Formulation Example 4 Flowable Agents: Thirty parts by weight of the compound of the present invention, 5 parts by weight of Solpol 9047K, 3 parts by weight of Sorbon T-20 (trade name; nonionic surfactant; Toho Pharmaceutical Co., Ltd.), 8 parts by weight of ethylene glycol and 44 parts by weight of water were ground by wet process with Dinomill (manufactured by Sinmaru Enterprise, Co., Ltd.).\",\n \"Ten parts by weight of an aqueous 1% by weight solution of xanthan gum (naturally occurring polymer) were added to the resulting slurry mixture, followed by thorough mixing and grinding, to obtain flowable agents containing the active ingredient at a content of 20% by weight.\",\n \"Test examples of fungicides and bactericides, insecticides and miticides for agriculture and horticulture and floriculture are shown below, which contain the compound of the present invention as the active ingredient.\",\n \"However, the mode for carrying out the present invention is not limited to those described below.\",\n \"Test Example 1 Fungicidal and Bactericidal Effect on Wheat Powdery Mildew: Compound (III) disclosed in JP-A-64-25763, tebufenpyrade and various compounds in accordance with the present invention were individually prepared into preparations in the same manner as in Formulation 3 The resulting preparations were diluted to given concentrations and sprayed on the stems and leave of a wheat plant (species: Norin No.61) at the one-leaf to 2-leaf stage as grown in a pot of a 6-cm diameter, at a ratio of 10-ml volume per one pot.\",\n \"After drying the chemical preparations in air, the wheat plant was exposed to the spore collected from wheat leave afflicted with Erypsiphe graminis as the microorganism causing wheat powdery mildew, for inoculation.\",\n \"Then, the plant was left to stand alone in a green house for 7 to 10 days.\",\n \"Evaluation was carried out follows.\",\n \"The ratio of the diseased area of each leaf was examined to calculate the control ratio according to the following formula.\",\n \"The results are shown in Table 3 (in the table, Compound Nos.\",\n \"correspond to Compound Nos.\",\n \"in Table-1.)\",\n \"Control ratio (%) =[(mean ratio of diseased areas in non-treated lot) −(mean ratio of diseased areas in treated lot)]× ×100 / (mean ratio of diseased areas in non-treated lot) TABLE 3 Compound No.\",\n \"Concentration (ppm) Control Ratio (%) 3 500 100 4 500 100 5 500 100 6 500 100 7 500 100 13 500 100 17 500 100 21 500 100 25 500 100 30 500 100 31 500 100 34 500 100 35 500 100 36 500 100 37 500 100 39 500 100 40 500 100 41 500 100 42 500 100 45 500 100 46 500 100 47 500 100 48 500 100 49 500 100 54 500 100 63 500 100 64 500 100 68 500 100 69 500 100 70 500 100 71 500 100 72 500 100 78 500 100 80 500 100 81 500 100 82 500 100 84 500 100 85 500 100 86 500 100 87 500 100 88 500 100 90 500 100 91 500 100 92 500 100 98 500 100 101 500 100 (III) 500 0 tebufenpyrade 500 77 Test Example 2 Fungicidal and Bactericidal Effect on Wheat Rust: Compound (III) disclosed in JP-A-64-25763 and various compounds in accordance with the present invention were individually prepared into preparations in the same manner as in Preparation 3.The resulting preparations were diluted to given concentrations and sprayed on the stems and leave of a wheat seedling (species: Norin No.61) at the one-leaf to 2-leaf stage as grown in a pot of a 6-cm diameter, at a ratio of 10-ml volume per one pot.\",\n \"After drying the chemical preparations in air, a spore suspension prepared by grinding the spore collected from wheat leave afflicted with Puccinia recondita as the microorganism causing wheat rust was sprayed on the wheat plant, for inoculation.\",\n \"Then, the plant was left to stand alone in a moist chamber at 22° C. for 15 hours.\",\n \"Subsequently, the plant was left to stand in a water pool in a green house for 7 days.\",\n \"Evaluation was carried out by examining the ratio of the diseased area of each leaf to calculate the control ratio by the same method as in Test Example 1.The results are shown in Table 4 (in the table, Compound Nos.\",\n \"correspond to Compound Nos, in Table-1.)\",\n \"TABLE 4 Compound No.\",\n \"Concentration (ppm) Control Ratio (%) 3 500 100 4 500 100 5 500 100 6 500 100 7 500 100 17 500 100 21 500 100 25 500 100 30 500 100 34 500 100 35 500 100 36 500 100 39 500 100 40 500 100 41 500 100 45 500 100 46 500 100 49 500 100 54 500 100 63 500 100 64 500 100 68 500 100 69 500 100 70 500 100 71 500 100 72 500 100 80 500 100 86 500 100 90 500 100 (III) 500 0 Test Example 3 Fungicidal and Bactericidal Effect on Larvae of Spodoptera litura: Compound (III) disclosed in JP-A-64-25763 and various compounds in accordance with the present invention were individually prepared into insecticide preparations according to the formulation of Formulation Example 1.The resulting insecticides of the present invention (hydrates) were diluted with water, where chopped cabbage leave (a diameter of 6 cm) were then immersed for one minute.\",\n \"After immersion and subsequent drying in air, the cabbage leave were placed in a plastic cup (inner diameter of 7 cm), where five larvae each of 3rd instar Spodoptera litura were left (twice in duplicate per one concentration).\",\n \"The plastic cup was kept in a thermostat at 25° C. Five days after the larvae were left in the plastic cup, the death and agony of these larvae were examined, to calculate the insecticide activity (%) while an agonizing insect was counted as 0.5 insect death.\",\n \"The results are shown in Table 5 (in the table, Compound Nos.\",\n \"correspond to Compound Nos.\",\n \"in Table-1.)\",\n \"TABLE 5 Compound No.\",\n \"Concentration (ppm) Insecticide Activity (%) 3 500 100 5 500 100 6 500 100 7 500 100 15 500 100 37 500 100 39 500 100 41 500 100 42 500 100 47 500 100 49 500 100 52 500 100 55 500 100 70 500 100 78 500 100 86 500 100 90 500 100 100 500 100 107 500 100 (III) 500 20 Test Example 4 Miticidal effect on adult Tetranychus urticae Koch The stem part of a Phaseolus seedling with one fresh first leaf remaining thereon was inserted in a test tube (a volume of 50 ml) containing water, where 15 female adult Tetranychus urticae Koch per one leaf was then inoculated.\",\n \"One day after the inoculation, the leave deposited with Tetranychus urticae Koch were immersed in and treated with an aqueous dilution of each of miticides (emulsions) individually produced from Compound (III) disclosed in JP-A-64-25763, tebufenpyrade and various compounds of the present invention according to the formulation for Formulation Example 3 (for about 5 seconds) (single one concentration, duplicate) The resulting test tubes were kept in a thermostat at 25° C. Five days after the treatment, the number of the female adult Tetranychus urticae Koch was counted on the Phaseolus leave, to calculate the adult death ratio (%) based on the above results.\",\n \"The results are shown in Table-6 (Compound Nos.\",\n \"in the table below correspond to Compound Nos.\",\n \"in Table-1).\",\n \"TABLE 6 Compound No.\",\n \"Concentration (ppm) Adult death ratio (%) 2 500 100 4 500 100 5 500 100 6 500 100 7 500 100 11 500 100 17 500 100 18 500 100 22 500 100 30 500 100 31 500 100 32 500 100 33 500 100 35 500 100 36 500 100 37 500 100 38 500 100 39 500 100 12.5 100 3.1 100 0.8 100 40 500 100 41 500 100 42 500 100 43 500 100 46 500 100 47 500 100 48 500 100 49 500 100 50 500 100 52 500 100 53 500 100 55 500 100 63 500 100 68 500 100 69 500 100 70 500 100 71 500 100 78 500 100 79 500 100 80 500 100 81 500 100 82 500 100 83 500 100 84 500 100 85 500 100 86 500 100 87 500 100 88 500 100 96 500 100 98 500 100 100 500 100 104 500 100 105 500 100 106 500 100 107 500 100 108 500 100 (III) 500 15 tebufenpyrade 12.5 100 3.1 80 0.8 0 While the invention has been described in detail with reference to specific embodiments thereof, it will be apparent to one skilled in the art that various changes and modifications can be made therein without departing from the spirit and scope thereof.\",\n \"The present application is based on a Japanese patent application filed on Apr.\",\n \"6, 2001 (Japanese Patent Application 2001-108116), the entire contents of which are incorporated hereinto by reference.\",\n \"Industrial Applicability The pyrazolecarboxamides of the present invention have such great effects on the control of bacterial strains, hazardous insects and mites that the pyrazolecarboxamides function as great insecticides and miticides for agriculture, forestry and disease control.\",\n \"Additionally, the pyrazolecarboxamides of the present invention are promising as a control agent of various hazardous biological organisms for livestock industry, fishery, or storage of various products and public hygiene.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10474179"}}},{"rowIdx":786,"cells":{"text":{"kind":"list like","value":[["Radio communication interface module, and its use in a device reading, processing, transmitting and operating on a bar code","A two-way radio communication interface module based on a predefined protocol for at least a user application, the module includes at least a radio circuit for transmitting and receiving digital signals and at least a programmable circuit with a radio microcontroller for managing the radio circuit.","The radio microcontroller includes a software stack consisting of a first part called protocol stack topped by a second part called applicative abstraction stack, the software stack enabling adaptation based on the protocol between the digital signals and the data and instructions of the application, the applicative abstraction stack enabling at least the translation of the data and instructions of the application into data and instructions based on the protocol for processing in the protocol stack."],["1-31.","(canceled) 32.Interface module (42) for two-way radiocommunication according to the <<Bluetooth®>> protocol for at least one user application having data and instructions to be communicated to an external device, the module including at least one radio electronic circuit (39, 40) for sending and receiving digital signals (43) and at least one programmable electronic circuit with a radio microcontroller (37) for managing the radio electronic circuit (39, 40), wherein the radio microcontroller (37) includes hardware means cooperating with software packages in the form of a software stack formed of a first section so-called protocol stack (38) topped with a second section so-called applicative abstraction stack (32), the software stack enabling adaptation according to the protocol between, on the one hand, the digital signals (43) and, on the other hand, the data and instructions of the user application, the applicative abstraction stack (32) enabling at least the translation of data and instructions of the user application into a corporate format in data and instructions according to the protocol for processing in the stack in order to enable the development of the application independently from the instructions and formats of the radiocommunication protocol, the module being a single hardware component also storing the user application in the form of a programme and ensuring its realization in the radio microcontroller (37), the software operation of the module being divided into tasks performed according to priorities, the tasks associated with the management of the radio circuit having higher priority than the execution tasks of the user application.","33.A module according to claim 32, wherein the applicative abstraction stack temporally manages the <<Bluetooth®>> connection mode, the interval of the running of the radio connection being modified according to the use of the module and in the course of time in order to manage the consumption of said module.","34.A module according to claim 33, wherein it includes, on the one hand, a radio printed circuit including an aerial, the radio electronic circuit for sending and receiving in the form of a radio and of a base band integrated circuit and the programmable electronic management circuit in the form of a microcontroller with storage means and, on the other hand, a printed circuit (41) interconnected with the radio printed circuit including at least one peripheral at least selected amongst: a digital input with possibly a comparator for level or transition detection, an analogue input with analogue-digital converter with possibly an analogue amplifier with programmable gain or not, a digital output, an analogue output with digital-analogue converter, a voltage controllable regulator or converter, an up/down counter, a real time clock, a charger circuit for battery, a logic circuit, a memory, a specialized programmable logic circuit notably selected amongst a microprocessor, a microcontroller, a digital signal processor or a dedicated circuit, a portion at least of the user application in the form of a programme being then stored in said specialized programmable logic circuit and said specialized programmable logic circuit ensuring then its realization, the applicative abstraction stack (32) may moreover comprise corporate software means for processing instructions for retrieving data from the peripheral and/or sending data to the peripheral.","35.A module according to claim 34, wherein it includes a reprogrammable memory as well as means enabling reprogrammation of said memory on the basis of data received by radiocommunication.","36.A module according to claim 35, wherein the user application is in the reprogrammable memory.","37.A module according to claim 36, wherein it is associated within a single box to an electrochemical supply source and at least one connector whereof the pins are linked with at least one interface of the module.","38.An application of the module according to claim 37 to the realization of symbol reading pen (1) support, the pen including in an optical sensor (2) a light source generating a light beam focused on the support and an optical detector intended to receive and convert the light reflected by the support into electric signals, the pen being stand-alone and including an internal electric power supply, the user application in the form of an operating programme including means for processing the electric signals of the optical detector and interfacing means via the radiocommunication module ensuring at least the transmission of said symbols according to said protocol to a data-processing equipment.","39.An application according to claim 38, wherein the symbols are barcodes.","40.An application according to claim 37 to the realization of a writing recognition reading pen, the pen including at least two accelerometers according to two axes X and Y corresponding to the writing plane and generating electric signals, the pen being stand-alone and including an internal electric power supply, the user application in the form of an operating programme including means for processing the electric signals and recognizing the writing, and interfacing means via the radiocommunication module ensuring at least the transmission of said writing according to said protocol to a data processing equipment.","41.An application according to claim 37 to the mains electric supply socket controlled by radiocommunication.","42.A method for reading, processing, transmitting and using a symbol on a support with a reading symbols pen according to claim 39, wherein: the symbol is read with the pen, said electric signals are processed in the interface module to generate digital and/or alpha-digital data, said digital and/or alpha-digital data are associated with a reader identifier in order to generate a chain of corporate data, and the chain of corporate data is transmitted in real time to at least one customer equipment including at least a software to carry out a command understandable by said customer equipment.","43.A method according to claim 42, wherein when a customer equipment is not available for reception, the chain of corporate data is stored in a memory of the reading pen.","44.A method according to claim 43, wherein in the customer equipment one associates moreover to the chain of corporate data at least one piece of data of customer equipment type in order to build a computer address depending then on the symbol, on the reader identifier and on the data of equipment type.","45.A method according to claim 44, wherein one implements a correspondence table in order to associate a computer address to the chain of corporate data, possibly associated with at least one piece of data of customer equipment type, the correspondence table being situated in a computer server distinct from the customer equipment.","46.A method according to claim 44, wherein one implements a correspondence table in order to associate a computer address with the chain of corporate data, possibly associated with at least one piece of data of customer equipment type, and that the correspondence table is stored in the customer equipment by selection according to a criterion in a global correspondence table situated in a computer server distinct from the customer equipment, the criterion including at least the reader identifier.","47.A method according to claim 46, wherein one encodes according to an encoding algorithm the digital and/or alpha-digital data before associating it with the reader identifier.","48.A method according to claim 46, wherein the computer address associated with the chain of corporate data is that of a computer server relaying an information request to another computer server.","49.A method according to claim 48, wherein the symbol is a barcode.","50.A method according to claim 49, wherein the support of the symbol is a display screen and that the symbols are barcodes corresponding to a successive temporal chaining of dark and clear surfaces on a display screen.","51.A method according to claim 33, wherein it includes, on the one hand, a radio printed circuit including a reprogrammable memory as well as means enabling reprogrammation of said memory on the basis of data received by radiocommunication."],["This invention relates to a radiocommunication module interface, a device, applications whereof a method for reading, processing, transmitting and using a barcode such that the method implements the module in a stand-alone barcode reader with radio link.","The invention may be implemented generically in all the hardware and software applications including a radio link and, also, implementation more specifically in a particular application in the field of business promotion, of commerce and more generally of data exchange on a computer network with pieces of equipment capable of processing hypermedia data such as text, image, sound and/or video.","Barcode type identification graphic codes are known and largely used due to the robustness of their reading and decoding.","Several encoding standards have been defined as well as regards the one-dimension as the two-dimension barcodes.","These codes are read by electronic apparatuses including an optical reading head.","According to the barcode scanning method and the type of optical reading head implemented, one has two large types of apparatus: the contact reading apparatuses which require physical displacement between the optical reading head and the support whereon is provided the barcode and the apparatuses capable of operating without any relative displacement, the scanning being obtained by mechanical methods (revolving mirror for example), and/or electronic methods (matrix detector for example).","For the latter apparatus, diverse technologies may be implemented but in all cases, these are apparatuses more complex and costly to be made than the apparatuses of the first type whereof the reading pen which is one of the simplest apparatuses to be made since it only includes a static optical sensor and whereof the scanning of the barcode for reading lecture is performed by the user.","The reading pens known according to the state of the art are generally linked by an electric cable to an external electronic box itself connected to the keyboard of a computer.","Such an optical pen is therefore not stand-alone and is not easier to manipulate since a cable links them to data-processing equipment.","Moreover, this type of pen is of relatively high cost since the optical sensor includes moreover an optical element which is very hard, sapphire for example, and which may sustain repeated frictions on the support including the barcode while ensuring optical adaptation.","Besides in known applications, the barcodes which are read are essentially processed locally.","For example in the applications of the payment check-out type, the data corresponding with the barcodes is processed at the check-out properly speaking to retrieve the price and, possibly, a description of the item, or in a local server of the business site connected to the set of check-outs in order to ensure also the management of the stock.","The object of the invention is to provide a radiocommunication hardware and software means in the form of a module which may adapt simply to any user application and, in a particular case, to a method wherein a computer network linking pieces of computer equipment capable of executing hypermedia applications implements data from reading barcodes in order to generate visual, acoustic actions and/or other, possibly interactive actions, on a piece of customer computer equipment in relation to a barcode reader.","The term computer equipment in the context of the invention corresponds to any electronic device capable of processing hypermedia data and of being connected to the network computer and, for example, computer, micro-computer, personal assistant, telephone with WAP, GPRS, UMTS, etc.","functionalities.","Thanks to the particular application of the invention, the user may read a barcode with a reader and may receive quasi instantly a page of information regarding the object carrying the barcode, on any type of computer support connected to Internet (micro-computer, digital assistant, mobile telephone or others).","The object may be any current consumption object (carrying then a barcode of type EAN or UPC) or any other type of object and, for example: visit cards, journal article, invoice, bank statement.","The term barcode relates to a digital value or a character as well as several according to a standardised encoding or a corporate encoding (corporate barcode).","The invention relates therefore to an interface module for two-way radiocommunication according to a preset protocol for at least one user application having data and instructions to be communicated to an external device, the module including at least one radio electronic circuit for sending and receiving digital signals and at least one programmable electronic circuit with a radio microcontroller for managing the radio electronic circuit.","According to the invention, the radio microcontroller includes hardware means co-operating with software packages in the form of a software stack formed of a first section so-called protocol stack topped with a second section so-called applicative abstraction stack, the software stack enabling the adaptation according to the protocol between, on the one hand, the digital signals and, on the other hand, the data and instructions of the user application, the applicative abstraction stack enabling at least the translation of data and instructions of the user application into a corporate format into data and instructions according to the protocol for processing in the protocol stack, the module being a single hardware component also storing the user application in the form of a programme and ensuring its realisation in the radio microcontroller.","In various embodiments of the invention, the following means which might be combined according to all the technically possible combinations are employed: the module is composed of a radio printed circuit including an aerial, the radio electronic circuit for sending and receiving in the form of a radio and of a base band integrated circuit and the programmable electronic management circuit in the form of a microcontroller with storage means, the software operation of the module is divided into tasks performed according to priorities, the tasks associated with the management of the radio circuit having higher priority than the execution tasks of the user application, the module includes moreover a second printed circuit interconnected with the radio printed circuit with at least one peripheral at least selected amongst: a digital input with possibly a comparator for level or transition detection, an analogue input with analogue-digital converter with possibly an analogue amplifier with programmable gain or non, a digital output, an analogue output with digital-analogue converter, a voltage controllable regulator or converter, an up/down counter, a real time clock, a charger circuit for battery, a logic circuit, a memory, a specialised programmable logic circuit notably selected amongst a microprocessor, a microcontroller, a digital signal processor or a dedicated circuit, a portion at least of the user application in the form of a programme being then stored in said specialised programmable logic circuit and said specialised programmable logic circuit ensuring then its realisation, the applicative abstraction stack capable moreover of comprising corporate software means for processing instructions for retrieving data from the peripheral and/or sending data to the peripheral, the module includes a reprogrammable memory as well as means enabling reprogrammation of said memory on the basis of data received by radiocommunication, the protocol is <<Bluetooth®>> or IEEE802.15.1, the applicative abstraction stack ensures moreover and conversely the translation of information of the protocol stack towards the application; the single component is an integrated circuit; the module includes moreover at least one infrared two-way communication means and the applicative abstraction stack includes moreover software means for processing instructions and data for and of said infrared means; the module includes a voltage regulator enabling to supply said module from a 220V/110V power supply socket, a battery or any other power source and the applicative abstraction stack includes moreover software means for processing instructions and data for and of said regulator; the module includes moreover some memory intended for storing data; the microcontroller is of a type comprised between 8 and 16 bits, pins included, with a frequency clock smaller than 200 KHz; the microcontroller includes some memory enabling the execution of a user application programme, the programme application including a sequence of instructions and of data according to the corporate format; the memory is a pre-programmed read-only memory; the memory is a chargeable (reprogrammable) memory; the chargeable memory is charged by the programme application at each beginning of said application in the module; the chargeable memory is charged by the programme application once and for all at the manufacture of the module in said application; the chargeable memory is charged by the programme application once and for all at the first start of said application in the module; the programme application for charging is in a read-only memory external to the module; the charging is performed by a wire-link, preferably a serial link; the programme application for charging is in a computer means external to the module; the charging is performed by the radio link of the module.","The invention relates also to a particular application of the module according to one or several of the previous features in the form of an interfacing device for two-way radiocommunication according to a preset protocol for at least one software user application and including a module according to any of the previous features, and according to which the device includes moreover within a single box, an electrochemical supply source, and at least one connector whereof the pins are linked with at least one interface of the module.","The invention relates also to a particular application of the module as such or in the device according to one or several of the previous features possibly combined for the realisation of an electronic apparatus including a software user application communicating by two-way radiocommunication according to a preset protocol and including a module according to any of the previous features.","In particular embodiments the apparatus is: a apparatus which is a barcode reading pen of a support, the pen including at one end in an optical sensor a light source generating a light beam focused on the support and an optical detector intended to receive and convert the light reflected by the support into electric signals, the pen being stand-alone and including an internal electric power supply and the radiocommunication module interface, with a software stack according to the <<Bluetooth>>® protocol, the user application in the form of an operating programme including means for processing the electric signals of the optical detector, and interfacing means via the radiocommunication module ensuring at least the transmission of said barcodes according to said protocol to a data-processing equipment, (the transmission of the barcodes corresponds to the characters that the user application has read) as an alternative to the support, the barcodes correspond to a successive temporal chaining of dark and clear surfaces on a display screen, a apparatus which is a reading pen with recognition of alpha-digital characters of a support, the pen including at one end in an optical sensor a light source generating a light beam focused on the support and an optical detector intended to receive and convert the light reflected by the support into electric signals, the pen being stand-alone and including an internal electric power supply and the radiocommunication module interface, with a software stack according to the <<Bluetooth®>> protocol, the user application in the form of an operating programme including means for processing the electric signals of the optical detector and recognising said characters, and interfacing means via the radiocommunication module ensuring at least the transmission of said characters according to said protocol to a data-processing equipment, a apparatus which is a writing recognition reading pen, the pen including at least two accelerometers according to two axes X and Y corresponding to the writing plane and generating electric signals, the pen being stand-alone and including an internal electric power supply and the radiocommunication module interface, with a software stack according to the <<Bluetooth®>> protocol, the user application in the form of an operating programme including means for processing the electric signals of the optical detector and recognising the writing, and interfacing means via the radiocommunication module ensuring at least the transmission of said writing according to said protocol to a data-processing equipment, (the transmission of the writing corresponds to the characters which the user application has read) the writing recognition reading pen includes a means to detect according to an axis Z the resting time of the pen when writing on a support, the means to detect the resting time is a contactor or an accelerometer, an apparatus which is one of the previous pens and which does not includes any focusing tip made of hard matter, an apparatus which is a mains electric supply socket controlled by radiocommunication, the socket includes a voltage reducer for the power supply of the module, a controlled electronic on/off switch, the reducer is selected amongst a chopping power supply, a static transformer power supply, a reducer capacitor power supply, the on/off switch is selected amongst an electromagnetic relay, a triac, an electronic on/off switch with zero-crossover.","The invention relates also to a particular application of the module as such or in the device according to one or several of the previous features possibly combined in the form of a method for reading, processing, transmitting and using a barcode wherein: the barcode is read with a reader including a radiocommunication module interface according to one or several of the previous features and optical reading means of the barcode optical reading head type generating electric signals relative to the bars, said electric signals are processed in the interface module to generate digital and/or alpha-digital data, said digital and/or alpha-digital data are associated a reader identifier in order to generate a chain of corporate data, one transmits in real time the chain of corporate data to at least one customer equipment including at least one customer software to carry out a command understandable by said customer equipment, In various modes of embodiment of the method the following means possibly combined are employed: when customer equipment is not available for reception, one stores the chain of corporate data in a memory of the reader, in the customer equipment one associates moreover with the chain of corporate data at least one piece of data of customer equipment type in order to build a computer address depending then on the barcode, on the reader identifier and on the data of equipment type, one implements a correspondence table in order to associate a computer address with the chain of corporate data, possibly associated with at least one piece of data of customer equipment type, the correspondence table being situated in a computer server distinct from the customer equipment, one implements a correspondence table in order to associate a computer address with the chain of corporate data, possibly associated with at least one piece of data of customer equipment type, the correspondence table being stored in the customer equipment, (one also considers the case when the correspondence table is shared between the customer equipment and at least one routing server, and, for example in case when no correspondence is found in the table of the customer equipment, a distinct routing server is then used) one stores the correspondence table in the customer equipment by selection according to a criterion in a global correspondence table situated in a computer server distinct from the customer equipment, the criterion including at least the reader identifier, one implements a transmission means by electromagnetic waves in free space between the reader and the customer equipment with a communication protocol enabling at least to determine which are the pieces of customer equipment capable of communicating and, in case when several pieces of equipment were operational, said protocol enabling to select at least one of said pieces of equipment operational, one encodes according to an encoding algorithm the digital and/or alpha-digital data before associating it with the reader identifier, the computer address associated with the chain of corporate data is that of a computer server relaying an information request to another computer server, the barcode is a corporate barcode; the identifier of the reader is a single number on 48 bits of <<Bluetooth®>> address; the identifier of the reader is a pre-programmed reader number during manufacture or programmed at a later stage; the internal electric power supply source is of the rechargeable battery and/or electrochemical battery type; the reader includes a means for providing its electric power supply by the pickup element of the user; the reader includes a means for cutting its electric power supply by the pickup element of the user; the means for providing the power supply is an electromechanical on/off switch of the push-button type, the power supply is cut off when the user releases said push-button; the push-button is laid-out so that the user necessarily positions the reader (or pen) in a given orientation with respect to the barcode; the given orientation is such that the reader (or pen) is substantially vertical when reading; when the barcode is read without error, the power supply of the optical sensor is cut off but the radiocommunication module remains powered; when the barcode is read without error, the power supply of the optical sensor is cut off but the radiocommunication module remains powered in low consumption mode until all the digital and/or alpha-digital data and/or binary data stored in the memory of the pen are sent back to the pieces of customer computer equipment; the reader includes moreover a sound generator; the programme enables to decode current barcodes among which those of EAN and UPC types; the programme enables moreover to decode barcodes according to a corporate encoding.","The invention relates finally to a symbol reading pen as well as a method with their embodiments possibly combined according to all the technically possible combinations and corresponding to: a symbol reading pen on a support, the pen including at one end in an optical sensor a light source generating a light beam focused on the support and an optical detector intended to receive and convert the light reflected by the support into electric signals, the pen being stand-alone and including an internal electric power supply source, an interface for two-way radiocommunication according to a preset protocol for at least one user application having data and instructions to be communicated to an external device as well as optical reading means of the optical reading head type generating electric signals relative to symbols, the user application in the form of an operating programme including a symbol recognition means and means for processing the electric signals of the optical detector, interfacing means via the radiocommunication interface ensuring at least the transmission of said symbols according to said protocol to a data-processing equipment, the symbols are barcodes on a support, the symbols are characters on a support, the symbols correspond to a writing, as an alternative to the support, the symbols are barcodes corresponding to a successive temporal chaining of dark and clear surface on a display screen, in the case of a display screen, the light source is omitted, a method for reading, processing, transmitting and using a symbol wherein: one reads the symbols with a reader which is a symbol reading pen according to one or several of the previous features, said electric signals are processed in the interface module to generate digital and/or alpha-digital data, said digital and/or alpha-digital data are associated a reader identifier in order to generate a chain of corporate data, one transmits in real time the chain of corporate data to at least one customer equipment including at least one customer software to carry out a command understandable by said customer equipment, a method according to the previous feature such that when a customer equipment is not available for reception, the chain of corporate data is stored in a memory of the reader, a method according to the previous features such that in the customer equipment one associates moreover with the chain of corporate data at least one piece of data of customer equipment type in order to build a computer address depending then on the symbol, on the reader identifier and on the data of equipment type, a method according to the previous features such that one implements a correspondence table in order to associate a computer address with the chain of corporate data, possibly associated with at least one piece of data of customer equipment type, the correspondence table being situated in a computer server distinct from the customer equipment, a method according to the previous features such that one implements a correspondence table in order to associate a computer address with the chain of corporate data, possibly associated with at least one piece of data of customer equipment type, the correspondence table being stored in the customer equipment, a method according to the previous features such that one stores the correspondence table in the customer equipment by selection according to a criterion in a global correspondence table situated in a computer server distinct from the customer equipment, the criterion including at least the reader identifier, a method according to the previous features such that one implements a transmission means by electromagnetic waves in free space between the reader and the customer equipment with a communication protocol enabling at least to determine which are the pieces of customer equipment capable of communicating and, in case when several pieces of equipment were operational, said protocol enabling to select at least one of said pieces of equipment operational, a method according to the previous features such that one encodes according to an encoding algorithm the digital and/or alpha-digital data before associating it with the reader identifier, a method according to the previous features such that the computer address associated with the chain of corporate data is that of a computer server relaying an information request to another computer server, a method according to the previous features such that the symbols are barcodes on a support, a method according to the previous features such that the symbols are characters on a support, a method according to the previous features such that the symbols are barcodes corresponding to a successive temporal chaining of dark and clear surfaces on a display screen.","This invention will be better understood when reading non limiting embodiments in relation to: FIG.","1 representing the general architecture (hardware and software) of a Smart Module, FIGS.","2 and 3 representing a reading pen according to two plans of visualisation, FIG.","4 representing the functional flowchart of the pen, FIG.","5 represents the reading pen in its environment, FIGS.","6 and 7a, 7b, 7c representing the operating steps of the pen, FIG.","8, representing schematically means for implementing of the method.","It should be reminded that, generally speaking, the software stack of the radiocommunication module interface includes a portion corresponding to the communication protocol (pile protocol) and a portion corresponding to the applicative abstraction stack (<<protocol abstraction API>>), the latter enabling simplified implementation of applications.","The applicative abstraction stack may correspond to one or several software levels.","The radiocommunication module interface, also designated <<SmartModule>> <<Baracoda®>> is a generic radio module in the form of a single hardware component which enables to carry software on-board user specific applications.","As a whole, a <<Smart Module>>, represented schematically on the FIG.","1, is composed of a radio means such as radio component, base band integrated circuit, as well as software means on-board a radio microcontroller, and as the case may be, of an aerial and/or of peripherals whereof, for example, analogue or digital inputs/outputs, a battery ensuring the autonomy of the module.","The <<SmartModule>> or radiocommunication module is a generic radio module which enables to carry on-board the application software.","The <<SmartModule>> is suited to all types of radio protocol, corporate or not and, for example, <<Bluetooth®>>, GPRS, CDMA2000, WCDMA, <<Home RF>>, 802.11b, <<WLAN>>, <<Wi-Fi>>, 802.11.a, <<Hyperlan>>, <<Zigbee>> or 433 MHz solutions.","The microcontroller of the interface module integrates all the software layers of the protocol in the protocol stack, the applicative abstraction stack, the profiles as well as on-board software (user application).","The applicative abstraction stack which may also process instructions and corporate data, enables, for example, fine management of the power supply and low consumption modes specified in the “Bluetooth®>> standard.","It enables as a whole to create an abstraction layer above the protocol stack corresponding to the protocol considered in order to provide software interface between the software specific to the protocol considered (the Bluetooth protocol stack, for example) and the user software, on-board.","This interface enables, as regards the user application, to simplify the use of the protocol and to vastly facilitate the development of on-board applications using an interface radio which is complex and very specific.","This enables therefore customers to accelerate the development without knowing the subtleties of the protocol used.","The peripherals associated (hardware), as well as their software management (master) enable, just as the on-board software, to facilitate the development of applications and the integration of the module in any type of electronic product.","Several peripherals are offered in a component library they may be for example voltage regulators (3.3V-5V), analogue/digital converters (ADC), digital/analogue converters (DAC), counters, comparators (at input), voltage converters, a battery charging circuit, an additional 8-bit microcontroller, basic software functions as well as inputs/outputs, for example, controlling an “LED” (light-emitting diode) or “buzzer” and reading the condition of an on/off switch or of an optical reader.","These peripherals may be added or removed during the electronic integration phase of the application.","In relation to the peripherals adopted or not, means for processing data and instructions of these peripherals will be or will not be inserted in the applicative abstraction stack.","In a particular version, the module includes a voltage regulator enabling to supply said module from a 220V/110V power supply socket, a battery or of any other power source.","The interest of these peripherals is that they do not modify the electronic functionalities of the product wherein one integrates the <<SmartModule>>: the applications introduced do not require any modification of the electronic board of the existing product, hence a gain in cost and integration time.","The selection of the different peripherals is made very quick by the use of a development kit <<SmartModule>> in the form of an electronic board whereon are welded all the peripherals which may be contemplated.","In a basic version, the peripherals are not connected but it is possible to position electric wires or jumpers between the inputs/outputs of each peripheral and the microcontroller.","From the position of the electric wires in the development board, one deduces therefrom the electric pattern of the module.","A specific routing and cabling work of the printed circuit is then necessary.","In a more evolved version, peripherals are addressable specifically through programmable logic or analogue gates and the functional configuration of the module is programmable instead of (or in addition to) the cabling phase.","In particular, the microcontroller may, for example, comprise an ADC and a counter.","It can be understood that the implementation of a particular peripheral relates as well to the hardware electronic circuit as to the associated software means in order to operate said peripheral and to recognise and to run corporate instructions which are specific thereto.","As already seen, the software means of the module may be pre-programmed or charged at usage.","In a particular case of protocol, <<Bluetooth>>® corresponds to a standard technology of short-range communication radio between two pieces of equipment.","The term customer equipment signifies computer equipment (electronic assistant, computer or mobile telephone) known by the reader.","They are pieces of equipment whereof the <<Bluetooth®>> address (48 bits) is recorded in a memory of the reader.","The object of the <<SmartModule>> is to facilitate the integration of the <<Bluetooth®>> technology in all types of electronic products, with a wire structure or not.","The radiocommunication module facilitates vastly the integration of said technology in electronic products since the conventional <<Bluetooth®>> devices do not integrate any on-board software and are controlled from a computer or processor external to the module (<<host>>), which requires the implementation between the device and the <<host>> of the instructions and data which are complex and specific to said technology.","However, in a conventional electronic product, the processor of said product does not contain the code which enables to control the <<Bluetooth®>> device and, moreover, it is very difficult to modify the code existing in an on-board processor because of the memory size and the absence of operating system in most rustic electronic products.","The Smart Module is therefore composed of a module hardware and of a module software.","FIG.","1 represents the pattern of the architecture of the Smart Module.","Hardware Architecture: The example of <<SmartModule>> represented is composed of 2 printed circuits, welded on one another.","The first printed circuit is the radio circuit properly speaking, it is composed of a aerial, of a radio, of a base band integrated circuit, of a microprocessor or microcontroller (<<radio MCU>>), of a FLASH or ROM-type programme memory and of a RAM-type memory.","It is called here radio module (RM).","The second printed circuit integrates at least all the peripherals necessary to the correct hardware operation of the radio module in its environment.","It is called here hardware connection (Hardware Patch or HWP).","Said second printed circuit may also comprise other peripherals as explained previously and which depend on the application contemplated.","Still, as explained previously, one or several interfaces not used in a particular application may be present by default, if only because they are integrated by default in the electronic circuits implemented.","In practice, because of the costs, several types of modules are provided including a variable number of interfaces and/or in relation to potential applications.","For example, a generic module intended for the study and the development of applications may comprise a large number of interfaces as well the modules intended for the large series production will include a minimum number or simply the interfaces necessary to the application.","In other versions, the radiocommunication module is on a single printed circuit and in a high integration version, the radiocommunication module is a single integrated circuit which is hybrid or, preferably, monolithic.","Software Architecture: The software corresponding to the stack with the protocol stack and the applicative abstraction stack and the possible applications of the user, is on-board the microcontroller(s) of the <(SmartModule)).","By default and in the minimum version of the module, there is a microcontroller (microprocessor, FLASH or ROM memory, RAM memory) on the radio circuit and this microcontroller is designated the <<radio MCU>>.","In more sophisticated versions of the module, it is possible to add a second microcontroller (8-bit, 16-bit, 32-bit or DSP) on the HWP, i.e.","the second printed circuit in the example represented.","The interest of a second microcontroller (external MCU) lies in the possibility of carrying out specific processes which could not be contemplated on the microcontroller of the radio circuit because of real time constraints, of limitation of the calculation power of the controller of the radio circuit, of the necessity to have additional and/or specific inputs/outputs (not available on the microcontroller of the radio circuit).","The following description provides a particular example of embodiment of the radiocommunication module within the framework of the <<Bluetooth®>>, protocol.","The <<SmartModule®>> facilitates the integration of the <<Bluetooth®>> technology in all types of electronic products, with a wire structure or not and enables to accelerate the development cycle of a new product.","The radio circuit is generic and there are 2 versions for the <<Bluetooth®>> protocol, a first short-range version with a maximum transmission power of 4 dBm (2.5 mW) and a second long-range version with a maximum transmission power of 20 dBm (100 mW).","The <<Radio MCU>> The <<radio MCU>> is a microcontroller using the processor ARM7 TDMI.","Other architectures 32-bit RISC can still be used.","The <<radio MCU>> has a ROM or FLASH-type programme memory.","This programme memory may be integrated on the chip of the processor or on an external integrated circuit and, in the latter case, the memory is in the radio module (RM).","A hybrid architecture may also be implemented.","The order of magnitude of the size of this ROM/FLASH memory is 1 Mbit to 32 Mbit, where this parameter may be led to change relative to technologic evolutions and/or memory requirements of the application.","A RAM memory is also on-board the radio module (RM).","It has a size of the order of 16 KBytes to 128 Kbytes, parameter depending on the technologic evolutions and/or memory requirements of the application.","A memory internal to the microcontroller and an external memory may be associated in the radio module (RM) in case when the memory requirements are significant, as well as for the ROM/FLASH as for the RAM.","Counters (<<timer>>) are also integrated in the <<radio MCU>>.","The Protocal Stack The portion of the software stack corresponding to the management of the protocol Bluetooth, stack protocol, is in this example, the <<Bluetooth® protocol Stack>>, includes the lower layers of the software (beneath the interface HCl), the higher layers (L2CAP, SDP, RFCOMM) as well as the <<profiles>>, above the higher layers of Bluetooth.","The <<Bluetooth®>> profiles are just beneath the applicative translation unit (applicative abstraction layer), itself beneath the possible applications of the user.","The Applicative Abstraction Stack This applicative abstraction stack (<<protocol abstraction-API>>) is arranged functionally above the layers or levels of the stack protocol, enabling a user application to use services of all the layers of the <<Bluetooth®>> protocol and to benefit from the wealth of this standard and, as already seen, possible additional local functionalities (local interface management, specific instructions .",".",".",").","This portion of the software stack is partially common to all the <<Smart Modules>> used in data acquisition tools (barcode reading, magnetic badge or contact-less reading, chip board reading, mouse, keyboard, temperature sensor, pressure sensor, movement sensor, writing recognition, CCD or CMOS camera, etc.",".",".",".",").","For other types of usage, there is provided another type of applicative abstraction stack.","In all cases, said stack enables to develop applications as simply as possible and independently from the instructions and formats of the communication protocol considered.","This applicative abstraction stack enables to access the radio interface without knowing the <<Bluetooth®>> protocol.","The user has three main commands: a data sending command which specifies the size of the data to be sent (2 bytes), the <<Bluetooth®>> address (48-bit MAC address) of the assignee (6 bytes), the profile used (1 byte), a service quality parameter (1 byte) and the data (<<payload>>).","a request command (<<inquiry>>) enabling the application <<Smart Module Manager>> situated above the applicative abstraction stack to find all the <<Bluetooth®>> products accessible in the vicinity and to recognise services that they offer.","listening to the <<Smart Module>> with a parameter (1 byte) characterising the listening mode.","These request commands (<<inquiry>>) and these listening mode activation commands are close to those offered by the GAP (<<General Access Profile>>).","Conversely, the first primitive send command is a very abstraction which masks the complexity of the <<Bluetooth®>> protocol and which enables a gain in development time.","We shall now detail the different parameters used in the send command: BDADR (<<Bluetooth®>> address of the assignee of the shipment): this address is provided by the <<Smart Module Manager>>.","Different addresses are stored in the non-volatile memory of the <<Smart Module>>, they come either from a configuration phase, or a request phase (<<inquiry>>).","The origin of the <<Bluetooth®>> addresses is not managed by the applicative abstraction stack, but at still higher a level by the <<Smart Module Manager>>.","the profile: one of the specificities of the <<Smart Module>> is to be able to manage a multi-terminal and multi-profile intelligent operation.","This aspect will be detailed in the particular case of the use of the <<Smart Module>> inside the barcode reading pen.","the service quality: this parameter defines whether an authentication is required, whether data should be encrypted, what usage type of the low consumption modes should be employed (arbitration level between consumption and delay for sending data).","<<Smart Module Manager>> It is a piece of software specific to the product wherein the Smart radiocommunication module is integrated.","This layer is truly the control layer of the <<Smart Module>>.","It is a high level software layer which enables to do away with the complexity of the radio protocol but also with the interconnection difficulties between the <<Smart Module>> and the product wherein it is integrated.","A portion of the <<Smart Module Manager>> manages the sending of data to one or several terminals twinned with the module in the configuration phase.","It therefore enables to manage a multi-terminal connection and to adapt the connection mode and the type of <<profile>> used in relation to the terminal.","During the configuration phase, several Bluetooth addresses of twinned terminals are recorded in a memory non-volatile of the Smart Module.","This layer is a corporate sub-layer developed by Baracoda.","It may be used in Smart Modules integrated in all types of data acquisition tools, such as barcode readers, among others.","The <<Smart Module Manager>> uses the services of the software HMI, <<Power Manager>>, <<IO Manager>> (inputs/outputs management) or any other type of specific application (decoding barcodes, encryption, form recognition algorithm, encoding/decoding of the video, voice, and image).","This specific application may also be a protocol stack (TCP/IP for example).","These vertical applications may operate on the radio microcontroller or on a specific external microcontroller if necessary.","The <<Smart Module Manager>> uses a number of software packages which are now described: MMI (<<Man Machine Interface>>): This software is specific to the environment wherein the <<Smart Module>> is used.","It enables to manage by a software the modification of the man/machine interface induced by the addition of a radio and of a battery.","The user must notably be informed of any successful shipment or not of data and/or when the battery is charged insufficiently and/or when it is in charging mode.","The man/machine interface is managed at the MMI layer of the <<Smart Module>>.","Hardware implementation of this software may be carried out either by using conventional peripherals such as LED, <<Buzzer>>, LCD screen or others, or, possibly associated, with new peripherals integrated on the HWP (battery charging circuit) to that end or at the terminal(s) with which the product is set up.","For example, the battery level of the product wherein the <<Smart Module>> is integrated may appear at the terminal and not on the product.","Power Manager (<<Power Manager>>): This software enables to use the Smart Module as power supply controller.","More precisely, this application software enables to control several power supply zones within the same product.","The purpose is to control by a software several regulators or transistors which enable to supply or not certain electric zones of the <<Smart Module>> properly speaking and also of the product wherein it is implemented.","These regulators and transistors are positioned on the hardware patch.","It should be noted that the battery charge management is not taken into account at the <<Power Manager>> software, but at the HWP (battery charge circuit).","Management of Inputs and Outputs (<<IO Manager>>): It is a layer which manages all the inputs/outputs between the product and the <<Smart module>>.","They are for example of the serial link type (synchronous or asynchronous) or parallel, analogue or digital.","On FIGS.","2 and 3, a pen 1 is seen in transparency.","Inside a box 10 of plastic material, an optical sensor 2 is arranged towards an end in relation to an optical channel 11 of hard material ahead of said pen 1.A communication module of Smart Module type is implemented in the form of a printed circuit 8 including an on/off switching means 3, push-button, a radio-electric component 4 according to the <<Bluetooth®>> protocol, a microcontroller 5 including read-only memory and random-access memory, whereas the memory may be one of the following types: ROM, EPROM, EEPROM, RAM, Flash, SRAM and an electric power supply source 6 in the form of two single-use batteries or of rechargeable batteries.","The optical sensor 2, the printed circuit 8, the power supply 6 are held in place by attachment means of the box 10.The pen as claimed is an application of the interface module claimed.","The Smart Module used, integrates a two-way radio component and a base band integrated circuit, according to the “Bluetooth®>> standard, as well as a microcontroller enabling to carry on-board, in whole or in part, the <<Bluetooth®>> protocol stack as well as dedicated software applications.","An additional microcontroller may be used for the listening phase, corresponding to the measurement of the successive lengths of a barcode.","The decoding is performed either on the external microcontroller or on the microcontroller on-board the <<Bluetooth®>> chip.","On FIG.","4, the previous pen is seen according to a plane perpendicular to the previous one enabling to observe on the printed circuit 8 a light-emitting diode (LED) 9 and an aerial 7 for the radio electric module 4.The aerial 7 is here an aerial of <<wire structure>> type printed on the printed circuit, but it might also be an aerial of <<patch>> type or any other type of radiating element placed inside the pen.","The two-colour LED 9 enables to inform the user on the operating condition of the pen.","This LED 9 may be replaced with any other type of alarm (sound generator, buzzer for example).","The optical sensor arranged at one end of the reader implements reflection optics with a light-emitting diode, for example of 700 nm wavelength, and a photodiode.","An aspheric forked lens or, in another embodiment not represented, a system of optical fibres, is arranged at the front of the light-emitting diode and of the photodiode in order to obtain a single focus point at the front of the pen.","The output signal of the optical sensor is a current generated in the photodiode and which is amplified by a transistor or an operational amplifier and which is converted into a binary voltage which may be processed by the microcontroller 5.A black surface absorbs the light emitted by the sensor and, in such a case, the output current is weak, the output is at a logic level 1.Conversely, outside a black surface, the light is reflected and the output voltage is at a logic level 0.The reader includes a box 10 and the optical sensor is held in the box at constant distance from the end of the pen, an optical channel 11 being realised between said end and the sensor.","Preferably, the optical channel is a light guide (optical fibre for example) or a lens (spherical tip) made of <<hard>> material.","The pen type reader of the invention is simple to make and includes preferably a box made of moulded plastic material and implements a reduced number of parts.","The electronic circuit of the reader includes a programme in a read-only memory intended for decoding barcodes of one of the following types: EAN, UPC, codes 128, code 39 and possibly specific barcodes.","Preferably, the specific barcodes comprise error correction means.","Preferably, the specific barcodes are encoded in order to comprise redundancies enabling, during decoding, to correct errors made when reading said barcode.","These corporate barcodes are encoded according to the principle of error correction codes used conventionally in the field of telecommunications.","Contrary to the state of the art, the pen claimed may read new types of barcodes on display screens, for example of television sets, computers or electronic assistants as well as screens of mobile telephones or any other type of screen using the plasma, TFT, liquid crystal (LCD) technology or by scanning (VGA, XVGA .",".",".",").","The barcodes used in such a case are very different from those of the state of the art: these are not codes having any <<spatial extension>> but rather a <<time extension>>.","More precisely, the barcodes claimed in the invention correspond to a succession of dark and clear surfaces on the display screen.","As a traditional barcode, the information is encoded in the successive durations of the bars and spaces of the code.","The user sees this code as a quick flashing of the screen.","As this code is decoded by the pen claimed, the user holds said pen in contact with the screen; whereas no displacement is performed on the screen.","This characteristic of the pen is a feature of the decoder (software module on-board the pen) and of the analogue circuit carrying out the conversion of the analogue signal from the photodiode, into a digital signal.","The main difficulty of this decoding type lies on the one hand in the low contrast between a <<white>> screen and a <<black>> screen.","On the other hand, in scanning-type systems, a refreshing frequency is used and complicates the decoding very much.","This characteristic being particular mainly to the decoding software, it is independent of the reading technology used (CCD, contact reading, CMOS sensor).","This example of reader in the form of a simplified reading pen which is stand-alone with low cost enables implementation of the method in everyday life and more especially in relation to computer means of the hypertext type (hypermedia) on micro-computer, personal assistant, mobile telephone, access point or others.","However, any type of reader including the functionalities specified may be implemented with the method of the invention.","The element common to the various reading technologies implemented is the interface module or Smart Module.","On FIG.","4 is represented a block diagram of the pen.","Said diagram enables among other things to integrate the on-board software and enables therefore to use the component Bluetooth, on the one hand to send and receive data by radio and on the other hand, to perform the decoding of the barcode, the management of the man/machine interface as well as the management of the power consumption and of the battery.","This enables to reduce the complexity (size of memory, calculation power) of the external microcontroller and hence to reduce the hardware cost of the product.","The original architecture (software and hardware) of the Smart Module, claimed in the invention, enables therefore to reduce very significantly the cost of a wireless reading pen and may therefore accelerate the implementation of the method in everyday life and more particularly in relation to computer means of the hypertext type (hypermedia) In a particular embodiment of the reader, for example the pen described, the push-button is arranged so that the user positions necessarily the reader in a given orientation with respect to the barcode.","The push-button is thus arranged so that the pickup element of the user holds the reader in a given position so that he may depress the button and launch the reading cycle.","This given orientation is such that the reader is substantially vertical, i.e.","substantially perpendicular with respect to the support of the barcode, when reading.","In case when the user should scan the barcode manually, the push-button is also arranged to facilitate said scanning and, for example, so that the pickup element of the user works according to scanning habits used when writing manually.","On FIG.","5, one has represented a pen 1 when reading a barcode 14 on an object 13.In the close environment which may be reached by radio-electric waves 18 of the module 4 several pieces of computer equipment are arranged which may communicate by electromagnetic waves according to the <<Bluetooth>>® protocol with said pen 1.These pieces of equipment are a micro-computer 15, a mobile telephone 16, with hypertext functionalities, a personal assistant 17 and one or several access points to a local network.","Initialisation and Configuration Phase A first initialisation phase is provided for the configuration of the pen.","The purpose of this phase is to specify which peripherals are <<twinned>> with the reader.","This initialisation method is managed at the processor on-board the Smart Module, at the software Smart Module Manager.","It comprises 4 steps: 1—Downloading the customer software on one of the user terminals (micro-computer for example).","This software is described schematically on FIG.","9.2—The user must then select the set of terminals with which he wants to <<twin>> the pen.","Two means are possible to fill in this list: or, the user performs, from the terminal, an <<inquiry>> and selects the terminals found by the inquiry.","This requires that the user configures all the terminals in listening mode (<<discoverable>>).","Or the user fills <<in long hand>> the list of Bluetooth addresses (48-bit MAC address).","Once the list of the terminals is formed, the user must then fill in a field containing the identifier number of the pen (48-bit address of the pen or any other identifier type of the pen).","The customer software then prompts the user to switch the pen into listening mode.","3—Several means are offered to the user to switch the pen into listening mode (<<discoverable mode>>).","The first corresponds to reading a specific barcode (corporate code) and the second method corresponds to the position of the pen in contact with a <<white>> paper for a duration of the order of 1 to 5s approximately.","These listening parameters of the pen may evolve and this description is not exhaustive.","In parallel, the customer software attempts a connection towards the pen and sends the information specific to the configuration: list of addresses of the peripherals or twinned terminals, the description of the <<profile>> to be used for each terminal, as well as an order of priority associated with each piece of equipment.","The NULL or BATCH terminal may be part of the list of the twinned terminals.","The purpose is to configure the reader in asynchronous mode: instead of trying to send, over radio, a decoded barcode towards a remote terminal, the pen stores said barcode in its non-volatile memory.","4—The configuration data is then stored in the non-volatile memory of the pen and might be modified at each instant by performing again a configuration phase.","Upon completed initialisation of the reader, the user is informed of the success of the initialisation phase via the man/machine interface of the pen and/or of the user terminal.","Once this initialisation phase has been conducted, the pen may be used as follows: On FIG.","6, only the telephone 17 is functional.","The link is then established between the equipment and the pen, the data read by the pen is read in the form of sets of digital or alpha-digital or binary data is transmitted by electromagnetic waves 20 towards the equipment 16 as represented on FIG.","3c.","The transmission of data is performed in real time, the data being transmitted as it is read and decoded or in quasi-real time, the handshake with the equipment or the pieces of equipment of the environment taking place before the transmission, the chains of data being stored temporally in the pen 1 as said link is being established.","Between reading 2 consecutive barcodes, the connection is held to maintain reduced consumption.","Management of Low Consumption Modes The power consumption is an essential element in the operation of the pen.","The <<Bluetooth®>> protocol has a great intrinsic advantage in terms of consumption with respect to other radio standards like 802.11b.","Nevertheless, the <<Bluetooth®>> protocol needs to be used in all its complexity and notably in terms of management of low consumption modes.","The <<Bluetooth®>> abstraction stack integrates this efficient management of the <<Bluetooth®>> connection modes.","When reading the barcode for the first time, the connection is created.","When the barcode is sent, the radio connection switches into <<sniff>> mode with a short <<Sniff>> interval.","This enables to keep a limited latency time.","After a certain time period (T1), the <<sniff>> period is increased in order to reduce the consumption still further.","The shortcoming in such a case is an increase in the lead time for sending a barcode.","It is nevertheless not a problem since, in such a case, the duration between 2 successive readings is long and the sending lead time is not fundamental.","After a certain time (T2), the reader switches into <<parked mode>> after exchanging the master/slave roles.","After a time T3, the radio connection is cut off and the power supply of the Smart Module is turned off.","The durations T1, T2, T3 are variable parameters which are adapted to the usage mode of the user and provide statistics of durations between successive readings.","For a given user reading barcodes at an average high rate, the values of T1, T2 and T3 will be selected as shorter.","Multi-Terminal Connection Management FIG.","5 represents a pen in its environment.","The pen may be set up with different and quite diverse pieces of equipment.","It is one of the significant elements of the “Bluetooth®” standard: interoperability and multi-terminal connectivity.","Nevertheless, even if these services are provided by the “Bluetooth®” standard, it is still necessary to manage this connectivity in an intelligent way taking into account the fact that the man/machine interface at the pen is very reduced and that it does not enable therefore to select towards which pieces of equipment the pen must be connected.","This intelligent management is a feature of the pen and is integrated in the applicative layer of the Smart Module.","During the configuration, the devices with which the pen may be connected are recorded in the non-volatile memory of the pen.","If the pen has been configured in asynchronous mode (by using only the NULL terminal in the configuration phase), each decoded barcode will be stored automatically in the non-volatile memory of the pen (FIG.","7a).","In order to access at a later stage the data recorded in the pen, the user must proceed to an upload or charging phase of the data.","This phase is performed by switching the pen into listening mode (by reading a corporate code for example) (FIG.","7b) while actuating the charging function on the customer software (micro-computer for example).","All the barcodes in memory are then transferred by radio waves towards the user terminal (FIG.","7c).","In the case of a real-time mode configuration (FIG.","6), i.e.","the pen has been configured with at least one non-NULL (or BATCH) user equipment, the pen tries to connect with different pieces of equipment in memory in order of priority or by using the equipment twinned last.","This handshake takes place only at the beginning of a reading phase.","A reading phase being characterised in that between 2 successive readings, the duration is shorter than T3 (duration of inactivity before total extinction of the power supply).","Throughout the reading phase, the connection is held in low consumption modes as described in the portion called <<management of low consumption modes>>.","If none of the pieces of equipment twinned reacts to the handshake from the pen, said pen is configured automatically (after one or several fruitless attempts) in asynchronous mode during the same reading phase.","The heterogeneity of the twinned terminals with a pen is very important.","The handshake modes, specified in the <<profiles>> of the <<Bluetooth®>> standard are very different of from one type of terminal to another.","The on-board software in the Smart Module of the pen is capable of managing several <<profiles>>.","More precisely, the applicative abstraction layer selects automatically the appropriate handshake mode in relation to the piece of equipment with which the pen wants to connect.","For example, in the case of an access point, the <<profile>> which is adequate is the <<Lan Access Point Profile>>, for a computer or digital assistant, it is the <<Serial Port Profile>> or <<HID profile>> and for a telephone it will be either the <<DialUp Networking Profile>>, by sending AT commands towards the telephone.","In such a case, the customer software is reduced to NULL on the terminal: the hypermedia navigator of the telephone being remote-controlled by the pen via AT commands.","Other solutions may be implemented, notably, it is possible to download a customer software on the SIM card of the mobile or in the processor memory of the telephone.","Battery Management The battery of the pen is rechargeable via an electronic circuit, integrated in the Smart Module.","Several voltage regulators, voltage controlled, are used on the Smart Module to control by a software, the power supply of different zones independent of the pen.","For example, when the user releases the action button, the zone corresponding to the reading head is not power supplied.","Conversely, the power supply of the radio module remains as long as the on-board software requests so.","This enables finer management of the consumption of the pen while separating correctly by functionalities the different electronic circuits forming the pen.","Man/Machine Interface Management This interface is managed by the software in the Smart Module of the pen.","On the pen there are light-emitting diodes or a sound generator.","This interface enables to signal the user a good reading or a bad reading, a successful radio send or not.","It also enables to inform the user when the battery level is too low.","A portion of this man/machine interface can be delocalised on the terminal when said terminal is provided with a richer interface (micro-computer or mobile telephone for example) In its application, the invention is structured around four components: the barcode reader, the customer equipment, the routing server, the document server, possibly reached by a relay server (proxy).","Anyone who wishes to access the information and services associated with a resource uses the barcode reader to scan the barcode referring to this resource.","The reader then decodes the barcode to generate a resource identifier.","Said identifier is sent with the own identifier of the reader towards the customer equipment, through a local connection.","The customer equipment processes the identifiers received to determine the address of an appropriate document server.","This server has the hypermedia documents exhibiting the information and services relative to the resource designated with the reader.","The customer equipment may, to facilitate its address research, obtain additional information from the routing server.","The identifiers from the reader are then relayed through a remote connection towards the document server, whereof the address has just been determined by the customer equipment.","Additional information whereof the user has authorised the broadcast may at this stage be transmitted over by the same channel.","Possibly, the remote connection towards the document server goes through a relay server or proxy, which is capable of storing the request and/or of processing it for statistical purposes or others, before transmitting it integrally to the document server.","Finally, the document server processes the data which has been transmitted thereto in order to send back to the customer equipment the hypermedia document which will enable the user to access the information and services requested.","All these steps take advantage of the latest technological breakthroughs in the field of telecommunications to offer real-time or quasi real-time service: there are only a few seconds between the scanning of the barcode and the presentation of the associated hypermedia document.","The barcode reader is a piece of hardware equipment capable of decoding a barcode into electric signals then into digital and/or alpha-digital data.","Such data forms an identifier of the resource referred to by the barcode.","This resource identifier and another identifier personal to the reader are both transmitted towards the customer equipment via a local connection.","The reader is intended for general public and consequently should be cheap enough.","It may conversely be of modest quality in comparison with industrial readers since its use is vastly less intensive.","it can be broken down into four main portions: an optical transceiver, a power supply, a local transmitter, a box.","The local connection is a short-range communication link between two pieces of equipment.","The distance between the pieces of equipment does not exceed a few ten metres.","Comprised as an electronic device, the local connection locale designates a short-range transceiver controlled via a communication protocol.","The customer equipment is the central element of the system.","The customer equipment is a piece of user equipment implementing the customer software.","The user equipment is a piece of hardware equipment possessed by the user.","It may have various characteristics, mobile or not, providing the following elements are available: a local connection, a remote connection, an operating system with a documented programming interface, a hypermedia navigator.","Concretely, a piece of mobile equipment may be a computer or a digital assistant connected to the Internet, a WAP mobile telephone offering programming possibilities, a GPRS, UMTS terminal or any other piece of equipment meeting the conditions detailed previously.","The remote connection is world-wide communication link between two pieces of equipment.","It almost always implements global networks, whereof the Internet is the best representative.","In this context, it may be contemplated to have all the requests transit via a set relay server (proxy), so that said relay updates the global information regarding the set of requests.","In all cases, the requests are at least transmitted integrally.","Comprised as an electronic device, the remote connection designates a network transceiver, possibly controlled via communication protocols.","The customer software is a piece of software running on any type of user equipment to fulfil the following functions: receive the identifiers transmitted by the barcode reader via the local connection, provide the user with the hypermedia document for accessing the information and services relative to the resource designated by the reader.","This piece of software is called customer software since it behaves as a customer for the routing server on the one hand and for the document server whereof it must determine the address on the other hand.","It is realised as an application programme, for example using an object-oriented programming language such as C++ or Java.","It controls the user equipment via the programming interface specific to such equipment.","The installation of this piece of software on a piece of user equipment may require a programme to be downloaded, the insertion of an electronic board whereon it is implanted (chop card for mobile telephone, extension card for portable computer or digital assistant) or any other electronic or computerised method.","The customer software may be broken down into three modules: a local receiver, a router, a navigator manager.","The local receiver is a programme whereof the role consists in listening further for the local connection.","It reacts to any message from the barcode reader while activating the router and while transmitting to said reader the identifiers received from the reader.","This programme is running as long as the system is actuated.","Conversely, when no message from the reader is being processed, it is the single portion of the customer software in operation, which reduces the consumption of the resources of the user equipment.","The router is a programme receiving from the local receiver the resource identifier and the reader identifier.","Its role consists in applying to these identifiers the address resolution algorithm to attempt to determine the WEB address of a document server appropriate to the resource referred.","If such an address is obtained, the router adds according to a set syntax both previous identifiers, as well as certain information which the user may have chosen to provide on its equipment (for example certain preferences or restrictions).","The address thus completed locates without any ambiguity a hypermedia document exhibiting information and services relative to the resource identified via the reader.","The router then actuates the navigator manager and transmits to said manager the address of the document.","The address resolution algorithm is applied to the resource and reader identifiers.","This algorithm processes first of all the reader identifier.","If the identifier is known, i.e.","if it listed on a routing table (correspondence) stored on the user equipment and managed by the router, then the programme extracts from the same table the routing information regarding the reader.","The routing table may be realised with diverse data structures; it forms a very simplified system for database management.","The routing information may contain: a history of the previous requests with their result, whereof one will be able to extract the address of a document server; a single address of document server, for example in case when the reader is exclusively affiliated to such server or a relay server address, through which all the requests pertaining to a document server should transit; any other information peculiar to the reader and useful to the address resolution.","In a second stage, the address resolution algorithm processes the resource identifier and the routing information extracted during the first step.","In some cases, this data is sufficient to generate the address of the document server associated with the resource referred.","For example, it may be agreed that an identifier exhibiting certain similarities with another identifier processed during a previous request must be associated with the address which resulted from this request.","If is not possible to conclude with this single data, the router transmits to the routing server the reader identifier and the resource reader, via the remote connection.","This server returns to the router the address of the document server which it is waiting for, as well as possibly the address of a relay server (proxy) associated, and these addresses are stored in the routing table.","The address resolution algorithm has therefore enabled to determine the address of the document server with at most a remote access to the routing server.","It should be noted however that it may be necessary to refresh periodically the routing information, via additional requests transmitted to the routing server during possible remote access.","In case when the reader identifier is not known from the router, the behaviour of the latter depends on the operating mode wherein the user has placed its customer equipment.","In current mode, the resolution procedure is aborted and the message from the reader is therefore ignored.","This normal situation may for example crop up if a message radio-transmitted by a user's reader is received by the customer equipment of another user, who is not interested at all in his neighbour's intentions.","If, conversely, the user has placed its customer equipment in receiving mode of a new barcode reader, then the new reader identifier is transmitted towards the routing server via the remote connection, so that this server sends back to the router the routing information peculiar to the new reader.","The receiving mode enables therefore to record a new reader.","Such information is placed in a new entry to the routing table, and the processing of the request by the customer software is completed.","The navigator manager is a programme whereof the role consists in causing the hypermedia navigator available on the user equipment to provide the hypermedia document located by the address received from the router.","The user is then free to navigate at will from the document which is presented to him.","The navigator manager only controls the display of a document to the usual navigator of the user, who remains perfectly free to set it up at his discretion.","To illustrate the mechanism implemented, one may consider the example of a WEB navigator to whom the navigator manager will order the creation of a new window to display an HTML document.","The address of this document is provided to the navigator in the form of a URL complying with the HTTP protocol.","Another example consists in reproducing on a mobile telephone connected to the Internet a sound document, located by an address complying with the WAP protocol.","The routing server is the auxiliary information source of the <<Baracoda®>> system (denomination adopted for the set of components, methods and services linked with the invention).","The routing server is a piece of software running on one or several computers fitted with a remote connection to fulfil the following functions: manage and update the routing information peculiar to each barcode reader; manage and update the information enabling to associate the WEB address of a relay server with the WEB address of a document server; manage and update the information enabling to associate the WEB address of a document server to a resource identifier accompanied by a reader identifier; manage and update the code of the different programmes forming the customer software; listen further to the remote connection in order to react to the requests of a customer equipment by transmitting to said customer the information that he requests.","It behaves therefore as a server for each piece of customer equipment, whereas the number of customers may be quite high.","It responds to the different requests of the router, but also to installation requests from a piece of user equipment that has not been fitted with the customer software.","The software routing server may be a single software running on a single computer, or be distributed on several computers.","The network organisation of these machines may itself implement diverse architectures: a hierarchical architecture, according to which the machines situated at the lowest nodes hold local information and refer to higher rank machines to obtain more general information; a neighbouring architecture, each machine being at the same rank as its neighbours and working in co-operation with the latter to access largely distributed information; any other architecture deemed efficient to share the processing load and capable of improving the response time to requests.","The communication between the routing server and a piece of customer equipment may employ a corporate protocol specifically designed to that end or a public protocol (HTTP or WAP for example).","Different protocols may be selected according to the nature of the customer equipment (computer, digital assistant, mobile telephone, etc.).","The document server is the last element of the ‘<Baracoda®>> system.","The document server is a piece of software running on one or several computers fitted with a remote connection to fulfil the following functions: manage and update a large number of hypermedia documents showing the information and services relative to certain resources; listen further to the remote connection in order to react to the requests of a hypermedia navigator by transmitting to said navigator the document that he requests.","The routing tables stored on the customer equipment or on the routing server comprise not only the addresses of document servers but also the addresses of relay servers associated with the document servers.","The software component called “router” of the “customer software” is therefore capable of providing the component called “navigator manager” with an address based on that of the relay server, so that the latter filters all the requests send to a given document server.","The relay server only re-transmits the request to the document server which is the final assignee thereof.","But it may in passing memorise that a request has been made and perform statistical processing of all the requests thus relayed.","This additional service offered to certain document servers may constitute an added value for anyone whose wishes to know the look-up frequency based upon the barcodes, the total volume of requests, their time distribution, etc.","The document server is the property of a third party wishing to provide the documents relative to certain resources.","It is implemented and managed to the discretion of the owner.","For example it may be the server of an industrial company publishing its catalogue of products on the Internet.","However, it must in order to be integrated into the <<Baracoda®>> system, be capable of interpreting the address transmitted by the navigator manager via the navigator.","To guarantee such compatibility at the lowest possible cost, the address considered is in most cases in compliance with a public protocol.","It may be for example URL, HTTP or WAP.","In such a case, the upgrade of an existing WEB server only requires minor adaptation, for example the design of a single page of dynamic redirection.","The application of the radiocommunication module is not limited to the method for reading barcodes and it is possible to implement the module in other applications.","For example, the <<Bluetooth®>>—GPRS router is an electronic apparatus composed of a <<SmartModule>> and of a modem GSM, GPRS, EDGE or UMTS.","Said apparatus carries out the conversion of protocol between the <<Bluetooth®>> communication protocol and the GPRS, EDGE or UMTS protocol.","It enables for example <<Bluetooth®>> products to access a remote network (Internet network for example).","The applications of such apparatus are numerous in the field of house computerised appliances and the field of Internet access (ISP) for a residential user or a small company.","The apparatus consists of a radiocommunication module and of a GSM/GPRS/EDGE/UMTS radio with management of the physical layer of the GSM/GPRS/EDGE/UMTS protocol and of the protocol stack considered as well as of a specific software managing the quality of service and the sharing of the pass-band between the different users.","Up to 7 products fitted with the apparatus may use the services of the GPRS/UMTS simultaneously, via the same <<Bluetooth®>>—GPRS router.","More than 7 pieces of equipment (up to 255) may access these services via the same router, but not simultaneously.","These pieces of equipment may use a token protocol (developed on the processor of the router) in order to send or receive data regularly using the GPRS/UMTS connection.","Thanks to the apparatus which integrates the necessary hardware and software, the user does not need to manage the complexity of the routing software due to the necessity of sharing the pass-band in the ascending direction (from the <<Bluetooth®>>—GPRS access point towards the GPRS base station).","The <<Bluetooth®>>—GPRS router is also used as a <<Hub>> within the framework of the construction of a local <<Bluetooth®>> network.","One may also implement such router in house computerised appliances.","Refrigerators, alarms, radiators, lighting units, on/off switches, thermometers and others sensors (water, gas, electricity meters), television set, VCRs may be connected to a remote network via a <<Bluetooth®>>—GPRS access point which is this <<Bluetooth®>>—GPRS router.","In such an application of the router it is for example possible to activate the heating system of one's country house via its mobile telephone, remotely.","The router may also be applied for accessing Internet at home with PCs or PDAs (portable electronic assistant) or inside a car."]],"string":"[\n [\n \"Radio communication interface module, and its use in a device reading, processing, transmitting and operating on a bar code\",\n \"A two-way radio communication interface module based on a predefined protocol for at least a user application, the module includes at least a radio circuit for transmitting and receiving digital signals and at least a programmable circuit with a radio microcontroller for managing the radio circuit.\",\n \"The radio microcontroller includes a software stack consisting of a first part called protocol stack topped by a second part called applicative abstraction stack, the software stack enabling adaptation based on the protocol between the digital signals and the data and instructions of the application, the applicative abstraction stack enabling at least the translation of the data and instructions of the application into data and instructions based on the protocol for processing in the protocol stack.\"\n ],\n [\n \"1-31.\",\n \"(canceled) 32.Interface module (42) for two-way radiocommunication according to the <<Bluetooth®>> protocol for at least one user application having data and instructions to be communicated to an external device, the module including at least one radio electronic circuit (39, 40) for sending and receiving digital signals (43) and at least one programmable electronic circuit with a radio microcontroller (37) for managing the radio electronic circuit (39, 40), wherein the radio microcontroller (37) includes hardware means cooperating with software packages in the form of a software stack formed of a first section so-called protocol stack (38) topped with a second section so-called applicative abstraction stack (32), the software stack enabling adaptation according to the protocol between, on the one hand, the digital signals (43) and, on the other hand, the data and instructions of the user application, the applicative abstraction stack (32) enabling at least the translation of data and instructions of the user application into a corporate format in data and instructions according to the protocol for processing in the stack in order to enable the development of the application independently from the instructions and formats of the radiocommunication protocol, the module being a single hardware component also storing the user application in the form of a programme and ensuring its realization in the radio microcontroller (37), the software operation of the module being divided into tasks performed according to priorities, the tasks associated with the management of the radio circuit having higher priority than the execution tasks of the user application.\",\n \"33.A module according to claim 32, wherein the applicative abstraction stack temporally manages the <<Bluetooth®>> connection mode, the interval of the running of the radio connection being modified according to the use of the module and in the course of time in order to manage the consumption of said module.\",\n \"34.A module according to claim 33, wherein it includes, on the one hand, a radio printed circuit including an aerial, the radio electronic circuit for sending and receiving in the form of a radio and of a base band integrated circuit and the programmable electronic management circuit in the form of a microcontroller with storage means and, on the other hand, a printed circuit (41) interconnected with the radio printed circuit including at least one peripheral at least selected amongst: a digital input with possibly a comparator for level or transition detection, an analogue input with analogue-digital converter with possibly an analogue amplifier with programmable gain or not, a digital output, an analogue output with digital-analogue converter, a voltage controllable regulator or converter, an up/down counter, a real time clock, a charger circuit for battery, a logic circuit, a memory, a specialized programmable logic circuit notably selected amongst a microprocessor, a microcontroller, a digital signal processor or a dedicated circuit, a portion at least of the user application in the form of a programme being then stored in said specialized programmable logic circuit and said specialized programmable logic circuit ensuring then its realization, the applicative abstraction stack (32) may moreover comprise corporate software means for processing instructions for retrieving data from the peripheral and/or sending data to the peripheral.\",\n \"35.A module according to claim 34, wherein it includes a reprogrammable memory as well as means enabling reprogrammation of said memory on the basis of data received by radiocommunication.\",\n \"36.A module according to claim 35, wherein the user application is in the reprogrammable memory.\",\n \"37.A module according to claim 36, wherein it is associated within a single box to an electrochemical supply source and at least one connector whereof the pins are linked with at least one interface of the module.\",\n \"38.An application of the module according to claim 37 to the realization of symbol reading pen (1) support, the pen including in an optical sensor (2) a light source generating a light beam focused on the support and an optical detector intended to receive and convert the light reflected by the support into electric signals, the pen being stand-alone and including an internal electric power supply, the user application in the form of an operating programme including means for processing the electric signals of the optical detector and interfacing means via the radiocommunication module ensuring at least the transmission of said symbols according to said protocol to a data-processing equipment.\",\n \"39.An application according to claim 38, wherein the symbols are barcodes.\",\n \"40.An application according to claim 37 to the realization of a writing recognition reading pen, the pen including at least two accelerometers according to two axes X and Y corresponding to the writing plane and generating electric signals, the pen being stand-alone and including an internal electric power supply, the user application in the form of an operating programme including means for processing the electric signals and recognizing the writing, and interfacing means via the radiocommunication module ensuring at least the transmission of said writing according to said protocol to a data processing equipment.\",\n \"41.An application according to claim 37 to the mains electric supply socket controlled by radiocommunication.\",\n \"42.A method for reading, processing, transmitting and using a symbol on a support with a reading symbols pen according to claim 39, wherein: the symbol is read with the pen, said electric signals are processed in the interface module to generate digital and/or alpha-digital data, said digital and/or alpha-digital data are associated with a reader identifier in order to generate a chain of corporate data, and the chain of corporate data is transmitted in real time to at least one customer equipment including at least a software to carry out a command understandable by said customer equipment.\",\n \"43.A method according to claim 42, wherein when a customer equipment is not available for reception, the chain of corporate data is stored in a memory of the reading pen.\",\n \"44.A method according to claim 43, wherein in the customer equipment one associates moreover to the chain of corporate data at least one piece of data of customer equipment type in order to build a computer address depending then on the symbol, on the reader identifier and on the data of equipment type.\",\n \"45.A method according to claim 44, wherein one implements a correspondence table in order to associate a computer address to the chain of corporate data, possibly associated with at least one piece of data of customer equipment type, the correspondence table being situated in a computer server distinct from the customer equipment.\",\n \"46.A method according to claim 44, wherein one implements a correspondence table in order to associate a computer address with the chain of corporate data, possibly associated with at least one piece of data of customer equipment type, and that the correspondence table is stored in the customer equipment by selection according to a criterion in a global correspondence table situated in a computer server distinct from the customer equipment, the criterion including at least the reader identifier.\",\n \"47.A method according to claim 46, wherein one encodes according to an encoding algorithm the digital and/or alpha-digital data before associating it with the reader identifier.\",\n \"48.A method according to claim 46, wherein the computer address associated with the chain of corporate data is that of a computer server relaying an information request to another computer server.\",\n \"49.A method according to claim 48, wherein the symbol is a barcode.\",\n \"50.A method according to claim 49, wherein the support of the symbol is a display screen and that the symbols are barcodes corresponding to a successive temporal chaining of dark and clear surfaces on a display screen.\",\n \"51.A method according to claim 33, wherein it includes, on the one hand, a radio printed circuit including a reprogrammable memory as well as means enabling reprogrammation of said memory on the basis of data received by radiocommunication.\"\n ],\n [\n \"This invention relates to a radiocommunication module interface, a device, applications whereof a method for reading, processing, transmitting and using a barcode such that the method implements the module in a stand-alone barcode reader with radio link.\",\n \"The invention may be implemented generically in all the hardware and software applications including a radio link and, also, implementation more specifically in a particular application in the field of business promotion, of commerce and more generally of data exchange on a computer network with pieces of equipment capable of processing hypermedia data such as text, image, sound and/or video.\",\n \"Barcode type identification graphic codes are known and largely used due to the robustness of their reading and decoding.\",\n \"Several encoding standards have been defined as well as regards the one-dimension as the two-dimension barcodes.\",\n \"These codes are read by electronic apparatuses including an optical reading head.\",\n \"According to the barcode scanning method and the type of optical reading head implemented, one has two large types of apparatus: the contact reading apparatuses which require physical displacement between the optical reading head and the support whereon is provided the barcode and the apparatuses capable of operating without any relative displacement, the scanning being obtained by mechanical methods (revolving mirror for example), and/or electronic methods (matrix detector for example).\",\n \"For the latter apparatus, diverse technologies may be implemented but in all cases, these are apparatuses more complex and costly to be made than the apparatuses of the first type whereof the reading pen which is one of the simplest apparatuses to be made since it only includes a static optical sensor and whereof the scanning of the barcode for reading lecture is performed by the user.\",\n \"The reading pens known according to the state of the art are generally linked by an electric cable to an external electronic box itself connected to the keyboard of a computer.\",\n \"Such an optical pen is therefore not stand-alone and is not easier to manipulate since a cable links them to data-processing equipment.\",\n \"Moreover, this type of pen is of relatively high cost since the optical sensor includes moreover an optical element which is very hard, sapphire for example, and which may sustain repeated frictions on the support including the barcode while ensuring optical adaptation.\",\n \"Besides in known applications, the barcodes which are read are essentially processed locally.\",\n \"For example in the applications of the payment check-out type, the data corresponding with the barcodes is processed at the check-out properly speaking to retrieve the price and, possibly, a description of the item, or in a local server of the business site connected to the set of check-outs in order to ensure also the management of the stock.\",\n \"The object of the invention is to provide a radiocommunication hardware and software means in the form of a module which may adapt simply to any user application and, in a particular case, to a method wherein a computer network linking pieces of computer equipment capable of executing hypermedia applications implements data from reading barcodes in order to generate visual, acoustic actions and/or other, possibly interactive actions, on a piece of customer computer equipment in relation to a barcode reader.\",\n \"The term computer equipment in the context of the invention corresponds to any electronic device capable of processing hypermedia data and of being connected to the network computer and, for example, computer, micro-computer, personal assistant, telephone with WAP, GPRS, UMTS, etc.\",\n \"functionalities.\",\n \"Thanks to the particular application of the invention, the user may read a barcode with a reader and may receive quasi instantly a page of information regarding the object carrying the barcode, on any type of computer support connected to Internet (micro-computer, digital assistant, mobile telephone or others).\",\n \"The object may be any current consumption object (carrying then a barcode of type EAN or UPC) or any other type of object and, for example: visit cards, journal article, invoice, bank statement.\",\n \"The term barcode relates to a digital value or a character as well as several according to a standardised encoding or a corporate encoding (corporate barcode).\",\n \"The invention relates therefore to an interface module for two-way radiocommunication according to a preset protocol for at least one user application having data and instructions to be communicated to an external device, the module including at least one radio electronic circuit for sending and receiving digital signals and at least one programmable electronic circuit with a radio microcontroller for managing the radio electronic circuit.\",\n \"According to the invention, the radio microcontroller includes hardware means co-operating with software packages in the form of a software stack formed of a first section so-called protocol stack topped with a second section so-called applicative abstraction stack, the software stack enabling the adaptation according to the protocol between, on the one hand, the digital signals and, on the other hand, the data and instructions of the user application, the applicative abstraction stack enabling at least the translation of data and instructions of the user application into a corporate format into data and instructions according to the protocol for processing in the protocol stack, the module being a single hardware component also storing the user application in the form of a programme and ensuring its realisation in the radio microcontroller.\",\n \"In various embodiments of the invention, the following means which might be combined according to all the technically possible combinations are employed: the module is composed of a radio printed circuit including an aerial, the radio electronic circuit for sending and receiving in the form of a radio and of a base band integrated circuit and the programmable electronic management circuit in the form of a microcontroller with storage means, the software operation of the module is divided into tasks performed according to priorities, the tasks associated with the management of the radio circuit having higher priority than the execution tasks of the user application, the module includes moreover a second printed circuit interconnected with the radio printed circuit with at least one peripheral at least selected amongst: a digital input with possibly a comparator for level or transition detection, an analogue input with analogue-digital converter with possibly an analogue amplifier with programmable gain or non, a digital output, an analogue output with digital-analogue converter, a voltage controllable regulator or converter, an up/down counter, a real time clock, a charger circuit for battery, a logic circuit, a memory, a specialised programmable logic circuit notably selected amongst a microprocessor, a microcontroller, a digital signal processor or a dedicated circuit, a portion at least of the user application in the form of a programme being then stored in said specialised programmable logic circuit and said specialised programmable logic circuit ensuring then its realisation, the applicative abstraction stack capable moreover of comprising corporate software means for processing instructions for retrieving data from the peripheral and/or sending data to the peripheral, the module includes a reprogrammable memory as well as means enabling reprogrammation of said memory on the basis of data received by radiocommunication, the protocol is <<Bluetooth®>> or IEEE802.15.1, the applicative abstraction stack ensures moreover and conversely the translation of information of the protocol stack towards the application; the single component is an integrated circuit; the module includes moreover at least one infrared two-way communication means and the applicative abstraction stack includes moreover software means for processing instructions and data for and of said infrared means; the module includes a voltage regulator enabling to supply said module from a 220V/110V power supply socket, a battery or any other power source and the applicative abstraction stack includes moreover software means for processing instructions and data for and of said regulator; the module includes moreover some memory intended for storing data; the microcontroller is of a type comprised between 8 and 16 bits, pins included, with a frequency clock smaller than 200 KHz; the microcontroller includes some memory enabling the execution of a user application programme, the programme application including a sequence of instructions and of data according to the corporate format; the memory is a pre-programmed read-only memory; the memory is a chargeable (reprogrammable) memory; the chargeable memory is charged by the programme application at each beginning of said application in the module; the chargeable memory is charged by the programme application once and for all at the manufacture of the module in said application; the chargeable memory is charged by the programme application once and for all at the first start of said application in the module; the programme application for charging is in a read-only memory external to the module; the charging is performed by a wire-link, preferably a serial link; the programme application for charging is in a computer means external to the module; the charging is performed by the radio link of the module.\",\n \"The invention relates also to a particular application of the module according to one or several of the previous features in the form of an interfacing device for two-way radiocommunication according to a preset protocol for at least one software user application and including a module according to any of the previous features, and according to which the device includes moreover within a single box, an electrochemical supply source, and at least one connector whereof the pins are linked with at least one interface of the module.\",\n \"The invention relates also to a particular application of the module as such or in the device according to one or several of the previous features possibly combined for the realisation of an electronic apparatus including a software user application communicating by two-way radiocommunication according to a preset protocol and including a module according to any of the previous features.\",\n \"In particular embodiments the apparatus is: a apparatus which is a barcode reading pen of a support, the pen including at one end in an optical sensor a light source generating a light beam focused on the support and an optical detector intended to receive and convert the light reflected by the support into electric signals, the pen being stand-alone and including an internal electric power supply and the radiocommunication module interface, with a software stack according to the <<Bluetooth>>® protocol, the user application in the form of an operating programme including means for processing the electric signals of the optical detector, and interfacing means via the radiocommunication module ensuring at least the transmission of said barcodes according to said protocol to a data-processing equipment, (the transmission of the barcodes corresponds to the characters that the user application has read) as an alternative to the support, the barcodes correspond to a successive temporal chaining of dark and clear surfaces on a display screen, a apparatus which is a reading pen with recognition of alpha-digital characters of a support, the pen including at one end in an optical sensor a light source generating a light beam focused on the support and an optical detector intended to receive and convert the light reflected by the support into electric signals, the pen being stand-alone and including an internal electric power supply and the radiocommunication module interface, with a software stack according to the <<Bluetooth®>> protocol, the user application in the form of an operating programme including means for processing the electric signals of the optical detector and recognising said characters, and interfacing means via the radiocommunication module ensuring at least the transmission of said characters according to said protocol to a data-processing equipment, a apparatus which is a writing recognition reading pen, the pen including at least two accelerometers according to two axes X and Y corresponding to the writing plane and generating electric signals, the pen being stand-alone and including an internal electric power supply and the radiocommunication module interface, with a software stack according to the <<Bluetooth®>> protocol, the user application in the form of an operating programme including means for processing the electric signals of the optical detector and recognising the writing, and interfacing means via the radiocommunication module ensuring at least the transmission of said writing according to said protocol to a data-processing equipment, (the transmission of the writing corresponds to the characters which the user application has read) the writing recognition reading pen includes a means to detect according to an axis Z the resting time of the pen when writing on a support, the means to detect the resting time is a contactor or an accelerometer, an apparatus which is one of the previous pens and which does not includes any focusing tip made of hard matter, an apparatus which is a mains electric supply socket controlled by radiocommunication, the socket includes a voltage reducer for the power supply of the module, a controlled electronic on/off switch, the reducer is selected amongst a chopping power supply, a static transformer power supply, a reducer capacitor power supply, the on/off switch is selected amongst an electromagnetic relay, a triac, an electronic on/off switch with zero-crossover.\",\n \"The invention relates also to a particular application of the module as such or in the device according to one or several of the previous features possibly combined in the form of a method for reading, processing, transmitting and using a barcode wherein: the barcode is read with a reader including a radiocommunication module interface according to one or several of the previous features and optical reading means of the barcode optical reading head type generating electric signals relative to the bars, said electric signals are processed in the interface module to generate digital and/or alpha-digital data, said digital and/or alpha-digital data are associated a reader identifier in order to generate a chain of corporate data, one transmits in real time the chain of corporate data to at least one customer equipment including at least one customer software to carry out a command understandable by said customer equipment, In various modes of embodiment of the method the following means possibly combined are employed: when customer equipment is not available for reception, one stores the chain of corporate data in a memory of the reader, in the customer equipment one associates moreover with the chain of corporate data at least one piece of data of customer equipment type in order to build a computer address depending then on the barcode, on the reader identifier and on the data of equipment type, one implements a correspondence table in order to associate a computer address with the chain of corporate data, possibly associated with at least one piece of data of customer equipment type, the correspondence table being situated in a computer server distinct from the customer equipment, one implements a correspondence table in order to associate a computer address with the chain of corporate data, possibly associated with at least one piece of data of customer equipment type, the correspondence table being stored in the customer equipment, (one also considers the case when the correspondence table is shared between the customer equipment and at least one routing server, and, for example in case when no correspondence is found in the table of the customer equipment, a distinct routing server is then used) one stores the correspondence table in the customer equipment by selection according to a criterion in a global correspondence table situated in a computer server distinct from the customer equipment, the criterion including at least the reader identifier, one implements a transmission means by electromagnetic waves in free space between the reader and the customer equipment with a communication protocol enabling at least to determine which are the pieces of customer equipment capable of communicating and, in case when several pieces of equipment were operational, said protocol enabling to select at least one of said pieces of equipment operational, one encodes according to an encoding algorithm the digital and/or alpha-digital data before associating it with the reader identifier, the computer address associated with the chain of corporate data is that of a computer server relaying an information request to another computer server, the barcode is a corporate barcode; the identifier of the reader is a single number on 48 bits of <<Bluetooth®>> address; the identifier of the reader is a pre-programmed reader number during manufacture or programmed at a later stage; the internal electric power supply source is of the rechargeable battery and/or electrochemical battery type; the reader includes a means for providing its electric power supply by the pickup element of the user; the reader includes a means for cutting its electric power supply by the pickup element of the user; the means for providing the power supply is an electromechanical on/off switch of the push-button type, the power supply is cut off when the user releases said push-button; the push-button is laid-out so that the user necessarily positions the reader (or pen) in a given orientation with respect to the barcode; the given orientation is such that the reader (or pen) is substantially vertical when reading; when the barcode is read without error, the power supply of the optical sensor is cut off but the radiocommunication module remains powered; when the barcode is read without error, the power supply of the optical sensor is cut off but the radiocommunication module remains powered in low consumption mode until all the digital and/or alpha-digital data and/or binary data stored in the memory of the pen are sent back to the pieces of customer computer equipment; the reader includes moreover a sound generator; the programme enables to decode current barcodes among which those of EAN and UPC types; the programme enables moreover to decode barcodes according to a corporate encoding.\",\n \"The invention relates finally to a symbol reading pen as well as a method with their embodiments possibly combined according to all the technically possible combinations and corresponding to: a symbol reading pen on a support, the pen including at one end in an optical sensor a light source generating a light beam focused on the support and an optical detector intended to receive and convert the light reflected by the support into electric signals, the pen being stand-alone and including an internal electric power supply source, an interface for two-way radiocommunication according to a preset protocol for at least one user application having data and instructions to be communicated to an external device as well as optical reading means of the optical reading head type generating electric signals relative to symbols, the user application in the form of an operating programme including a symbol recognition means and means for processing the electric signals of the optical detector, interfacing means via the radiocommunication interface ensuring at least the transmission of said symbols according to said protocol to a data-processing equipment, the symbols are barcodes on a support, the symbols are characters on a support, the symbols correspond to a writing, as an alternative to the support, the symbols are barcodes corresponding to a successive temporal chaining of dark and clear surface on a display screen, in the case of a display screen, the light source is omitted, a method for reading, processing, transmitting and using a symbol wherein: one reads the symbols with a reader which is a symbol reading pen according to one or several of the previous features, said electric signals are processed in the interface module to generate digital and/or alpha-digital data, said digital and/or alpha-digital data are associated a reader identifier in order to generate a chain of corporate data, one transmits in real time the chain of corporate data to at least one customer equipment including at least one customer software to carry out a command understandable by said customer equipment, a method according to the previous feature such that when a customer equipment is not available for reception, the chain of corporate data is stored in a memory of the reader, a method according to the previous features such that in the customer equipment one associates moreover with the chain of corporate data at least one piece of data of customer equipment type in order to build a computer address depending then on the symbol, on the reader identifier and on the data of equipment type, a method according to the previous features such that one implements a correspondence table in order to associate a computer address with the chain of corporate data, possibly associated with at least one piece of data of customer equipment type, the correspondence table being situated in a computer server distinct from the customer equipment, a method according to the previous features such that one implements a correspondence table in order to associate a computer address with the chain of corporate data, possibly associated with at least one piece of data of customer equipment type, the correspondence table being stored in the customer equipment, a method according to the previous features such that one stores the correspondence table in the customer equipment by selection according to a criterion in a global correspondence table situated in a computer server distinct from the customer equipment, the criterion including at least the reader identifier, a method according to the previous features such that one implements a transmission means by electromagnetic waves in free space between the reader and the customer equipment with a communication protocol enabling at least to determine which are the pieces of customer equipment capable of communicating and, in case when several pieces of equipment were operational, said protocol enabling to select at least one of said pieces of equipment operational, a method according to the previous features such that one encodes according to an encoding algorithm the digital and/or alpha-digital data before associating it with the reader identifier, a method according to the previous features such that the computer address associated with the chain of corporate data is that of a computer server relaying an information request to another computer server, a method according to the previous features such that the symbols are barcodes on a support, a method according to the previous features such that the symbols are characters on a support, a method according to the previous features such that the symbols are barcodes corresponding to a successive temporal chaining of dark and clear surfaces on a display screen.\",\n \"This invention will be better understood when reading non limiting embodiments in relation to: FIG.\",\n \"1 representing the general architecture (hardware and software) of a Smart Module, FIGS.\",\n \"2 and 3 representing a reading pen according to two plans of visualisation, FIG.\",\n \"4 representing the functional flowchart of the pen, FIG.\",\n \"5 represents the reading pen in its environment, FIGS.\",\n \"6 and 7a, 7b, 7c representing the operating steps of the pen, FIG.\",\n \"8, representing schematically means for implementing of the method.\",\n \"It should be reminded that, generally speaking, the software stack of the radiocommunication module interface includes a portion corresponding to the communication protocol (pile protocol) and a portion corresponding to the applicative abstraction stack (<<protocol abstraction API>>), the latter enabling simplified implementation of applications.\",\n \"The applicative abstraction stack may correspond to one or several software levels.\",\n \"The radiocommunication module interface, also designated <<SmartModule>> <<Baracoda®>> is a generic radio module in the form of a single hardware component which enables to carry software on-board user specific applications.\",\n \"As a whole, a <<Smart Module>>, represented schematically on the FIG.\",\n \"1, is composed of a radio means such as radio component, base band integrated circuit, as well as software means on-board a radio microcontroller, and as the case may be, of an aerial and/or of peripherals whereof, for example, analogue or digital inputs/outputs, a battery ensuring the autonomy of the module.\",\n \"The <<SmartModule>> or radiocommunication module is a generic radio module which enables to carry on-board the application software.\",\n \"The <<SmartModule>> is suited to all types of radio protocol, corporate or not and, for example, <<Bluetooth®>>, GPRS, CDMA2000, WCDMA, <<Home RF>>, 802.11b, <<WLAN>>, <<Wi-Fi>>, 802.11.a, <<Hyperlan>>, <<Zigbee>> or 433 MHz solutions.\",\n \"The microcontroller of the interface module integrates all the software layers of the protocol in the protocol stack, the applicative abstraction stack, the profiles as well as on-board software (user application).\",\n \"The applicative abstraction stack which may also process instructions and corporate data, enables, for example, fine management of the power supply and low consumption modes specified in the “Bluetooth®>> standard.\",\n \"It enables as a whole to create an abstraction layer above the protocol stack corresponding to the protocol considered in order to provide software interface between the software specific to the protocol considered (the Bluetooth protocol stack, for example) and the user software, on-board.\",\n \"This interface enables, as regards the user application, to simplify the use of the protocol and to vastly facilitate the development of on-board applications using an interface radio which is complex and very specific.\",\n \"This enables therefore customers to accelerate the development without knowing the subtleties of the protocol used.\",\n \"The peripherals associated (hardware), as well as their software management (master) enable, just as the on-board software, to facilitate the development of applications and the integration of the module in any type of electronic product.\",\n \"Several peripherals are offered in a component library they may be for example voltage regulators (3.3V-5V), analogue/digital converters (ADC), digital/analogue converters (DAC), counters, comparators (at input), voltage converters, a battery charging circuit, an additional 8-bit microcontroller, basic software functions as well as inputs/outputs, for example, controlling an “LED” (light-emitting diode) or “buzzer” and reading the condition of an on/off switch or of an optical reader.\",\n \"These peripherals may be added or removed during the electronic integration phase of the application.\",\n \"In relation to the peripherals adopted or not, means for processing data and instructions of these peripherals will be or will not be inserted in the applicative abstraction stack.\",\n \"In a particular version, the module includes a voltage regulator enabling to supply said module from a 220V/110V power supply socket, a battery or of any other power source.\",\n \"The interest of these peripherals is that they do not modify the electronic functionalities of the product wherein one integrates the <<SmartModule>>: the applications introduced do not require any modification of the electronic board of the existing product, hence a gain in cost and integration time.\",\n \"The selection of the different peripherals is made very quick by the use of a development kit <<SmartModule>> in the form of an electronic board whereon are welded all the peripherals which may be contemplated.\",\n \"In a basic version, the peripherals are not connected but it is possible to position electric wires or jumpers between the inputs/outputs of each peripheral and the microcontroller.\",\n \"From the position of the electric wires in the development board, one deduces therefrom the electric pattern of the module.\",\n \"A specific routing and cabling work of the printed circuit is then necessary.\",\n \"In a more evolved version, peripherals are addressable specifically through programmable logic or analogue gates and the functional configuration of the module is programmable instead of (or in addition to) the cabling phase.\",\n \"In particular, the microcontroller may, for example, comprise an ADC and a counter.\",\n \"It can be understood that the implementation of a particular peripheral relates as well to the hardware electronic circuit as to the associated software means in order to operate said peripheral and to recognise and to run corporate instructions which are specific thereto.\",\n \"As already seen, the software means of the module may be pre-programmed or charged at usage.\",\n \"In a particular case of protocol, <<Bluetooth>>® corresponds to a standard technology of short-range communication radio between two pieces of equipment.\",\n \"The term customer equipment signifies computer equipment (electronic assistant, computer or mobile telephone) known by the reader.\",\n \"They are pieces of equipment whereof the <<Bluetooth®>> address (48 bits) is recorded in a memory of the reader.\",\n \"The object of the <<SmartModule>> is to facilitate the integration of the <<Bluetooth®>> technology in all types of electronic products, with a wire structure or not.\",\n \"The radiocommunication module facilitates vastly the integration of said technology in electronic products since the conventional <<Bluetooth®>> devices do not integrate any on-board software and are controlled from a computer or processor external to the module (<<host>>), which requires the implementation between the device and the <<host>> of the instructions and data which are complex and specific to said technology.\",\n \"However, in a conventional electronic product, the processor of said product does not contain the code which enables to control the <<Bluetooth®>> device and, moreover, it is very difficult to modify the code existing in an on-board processor because of the memory size and the absence of operating system in most rustic electronic products.\",\n \"The Smart Module is therefore composed of a module hardware and of a module software.\",\n \"FIG.\",\n \"1 represents the pattern of the architecture of the Smart Module.\",\n \"Hardware Architecture: The example of <<SmartModule>> represented is composed of 2 printed circuits, welded on one another.\",\n \"The first printed circuit is the radio circuit properly speaking, it is composed of a aerial, of a radio, of a base band integrated circuit, of a microprocessor or microcontroller (<<radio MCU>>), of a FLASH or ROM-type programme memory and of a RAM-type memory.\",\n \"It is called here radio module (RM).\",\n \"The second printed circuit integrates at least all the peripherals necessary to the correct hardware operation of the radio module in its environment.\",\n \"It is called here hardware connection (Hardware Patch or HWP).\",\n \"Said second printed circuit may also comprise other peripherals as explained previously and which depend on the application contemplated.\",\n \"Still, as explained previously, one or several interfaces not used in a particular application may be present by default, if only because they are integrated by default in the electronic circuits implemented.\",\n \"In practice, because of the costs, several types of modules are provided including a variable number of interfaces and/or in relation to potential applications.\",\n \"For example, a generic module intended for the study and the development of applications may comprise a large number of interfaces as well the modules intended for the large series production will include a minimum number or simply the interfaces necessary to the application.\",\n \"In other versions, the radiocommunication module is on a single printed circuit and in a high integration version, the radiocommunication module is a single integrated circuit which is hybrid or, preferably, monolithic.\",\n \"Software Architecture: The software corresponding to the stack with the protocol stack and the applicative abstraction stack and the possible applications of the user, is on-board the microcontroller(s) of the <(SmartModule)).\",\n \"By default and in the minimum version of the module, there is a microcontroller (microprocessor, FLASH or ROM memory, RAM memory) on the radio circuit and this microcontroller is designated the <<radio MCU>>.\",\n \"In more sophisticated versions of the module, it is possible to add a second microcontroller (8-bit, 16-bit, 32-bit or DSP) on the HWP, i.e.\",\n \"the second printed circuit in the example represented.\",\n \"The interest of a second microcontroller (external MCU) lies in the possibility of carrying out specific processes which could not be contemplated on the microcontroller of the radio circuit because of real time constraints, of limitation of the calculation power of the controller of the radio circuit, of the necessity to have additional and/or specific inputs/outputs (not available on the microcontroller of the radio circuit).\",\n \"The following description provides a particular example of embodiment of the radiocommunication module within the framework of the <<Bluetooth®>>, protocol.\",\n \"The <<SmartModule®>> facilitates the integration of the <<Bluetooth®>> technology in all types of electronic products, with a wire structure or not and enables to accelerate the development cycle of a new product.\",\n \"The radio circuit is generic and there are 2 versions for the <<Bluetooth®>> protocol, a first short-range version with a maximum transmission power of 4 dBm (2.5 mW) and a second long-range version with a maximum transmission power of 20 dBm (100 mW).\",\n \"The <<Radio MCU>> The <<radio MCU>> is a microcontroller using the processor ARM7 TDMI.\",\n \"Other architectures 32-bit RISC can still be used.\",\n \"The <<radio MCU>> has a ROM or FLASH-type programme memory.\",\n \"This programme memory may be integrated on the chip of the processor or on an external integrated circuit and, in the latter case, the memory is in the radio module (RM).\",\n \"A hybrid architecture may also be implemented.\",\n \"The order of magnitude of the size of this ROM/FLASH memory is 1 Mbit to 32 Mbit, where this parameter may be led to change relative to technologic evolutions and/or memory requirements of the application.\",\n \"A RAM memory is also on-board the radio module (RM).\",\n \"It has a size of the order of 16 KBytes to 128 Kbytes, parameter depending on the technologic evolutions and/or memory requirements of the application.\",\n \"A memory internal to the microcontroller and an external memory may be associated in the radio module (RM) in case when the memory requirements are significant, as well as for the ROM/FLASH as for the RAM.\",\n \"Counters (<<timer>>) are also integrated in the <<radio MCU>>.\",\n \"The Protocal Stack The portion of the software stack corresponding to the management of the protocol Bluetooth, stack protocol, is in this example, the <<Bluetooth® protocol Stack>>, includes the lower layers of the software (beneath the interface HCl), the higher layers (L2CAP, SDP, RFCOMM) as well as the <<profiles>>, above the higher layers of Bluetooth.\",\n \"The <<Bluetooth®>> profiles are just beneath the applicative translation unit (applicative abstraction layer), itself beneath the possible applications of the user.\",\n \"The Applicative Abstraction Stack This applicative abstraction stack (<<protocol abstraction-API>>) is arranged functionally above the layers or levels of the stack protocol, enabling a user application to use services of all the layers of the <<Bluetooth®>> protocol and to benefit from the wealth of this standard and, as already seen, possible additional local functionalities (local interface management, specific instructions .\",\n \".\",\n \".\",\n \").\",\n \"This portion of the software stack is partially common to all the <<Smart Modules>> used in data acquisition tools (barcode reading, magnetic badge or contact-less reading, chip board reading, mouse, keyboard, temperature sensor, pressure sensor, movement sensor, writing recognition, CCD or CMOS camera, etc.\",\n \".\",\n \".\",\n \".\",\n \").\",\n \"For other types of usage, there is provided another type of applicative abstraction stack.\",\n \"In all cases, said stack enables to develop applications as simply as possible and independently from the instructions and formats of the communication protocol considered.\",\n \"This applicative abstraction stack enables to access the radio interface without knowing the <<Bluetooth®>> protocol.\",\n \"The user has three main commands: a data sending command which specifies the size of the data to be sent (2 bytes), the <<Bluetooth®>> address (48-bit MAC address) of the assignee (6 bytes), the profile used (1 byte), a service quality parameter (1 byte) and the data (<<payload>>).\",\n \"a request command (<<inquiry>>) enabling the application <<Smart Module Manager>> situated above the applicative abstraction stack to find all the <<Bluetooth®>> products accessible in the vicinity and to recognise services that they offer.\",\n \"listening to the <<Smart Module>> with a parameter (1 byte) characterising the listening mode.\",\n \"These request commands (<<inquiry>>) and these listening mode activation commands are close to those offered by the GAP (<<General Access Profile>>).\",\n \"Conversely, the first primitive send command is a very abstraction which masks the complexity of the <<Bluetooth®>> protocol and which enables a gain in development time.\",\n \"We shall now detail the different parameters used in the send command: BDADR (<<Bluetooth®>> address of the assignee of the shipment): this address is provided by the <<Smart Module Manager>>.\",\n \"Different addresses are stored in the non-volatile memory of the <<Smart Module>>, they come either from a configuration phase, or a request phase (<<inquiry>>).\",\n \"The origin of the <<Bluetooth®>> addresses is not managed by the applicative abstraction stack, but at still higher a level by the <<Smart Module Manager>>.\",\n \"the profile: one of the specificities of the <<Smart Module>> is to be able to manage a multi-terminal and multi-profile intelligent operation.\",\n \"This aspect will be detailed in the particular case of the use of the <<Smart Module>> inside the barcode reading pen.\",\n \"the service quality: this parameter defines whether an authentication is required, whether data should be encrypted, what usage type of the low consumption modes should be employed (arbitration level between consumption and delay for sending data).\",\n \"<<Smart Module Manager>> It is a piece of software specific to the product wherein the Smart radiocommunication module is integrated.\",\n \"This layer is truly the control layer of the <<Smart Module>>.\",\n \"It is a high level software layer which enables to do away with the complexity of the radio protocol but also with the interconnection difficulties between the <<Smart Module>> and the product wherein it is integrated.\",\n \"A portion of the <<Smart Module Manager>> manages the sending of data to one or several terminals twinned with the module in the configuration phase.\",\n \"It therefore enables to manage a multi-terminal connection and to adapt the connection mode and the type of <<profile>> used in relation to the terminal.\",\n \"During the configuration phase, several Bluetooth addresses of twinned terminals are recorded in a memory non-volatile of the Smart Module.\",\n \"This layer is a corporate sub-layer developed by Baracoda.\",\n \"It may be used in Smart Modules integrated in all types of data acquisition tools, such as barcode readers, among others.\",\n \"The <<Smart Module Manager>> uses the services of the software HMI, <<Power Manager>>, <<IO Manager>> (inputs/outputs management) or any other type of specific application (decoding barcodes, encryption, form recognition algorithm, encoding/decoding of the video, voice, and image).\",\n \"This specific application may also be a protocol stack (TCP/IP for example).\",\n \"These vertical applications may operate on the radio microcontroller or on a specific external microcontroller if necessary.\",\n \"The <<Smart Module Manager>> uses a number of software packages which are now described: MMI (<<Man Machine Interface>>): This software is specific to the environment wherein the <<Smart Module>> is used.\",\n \"It enables to manage by a software the modification of the man/machine interface induced by the addition of a radio and of a battery.\",\n \"The user must notably be informed of any successful shipment or not of data and/or when the battery is charged insufficiently and/or when it is in charging mode.\",\n \"The man/machine interface is managed at the MMI layer of the <<Smart Module>>.\",\n \"Hardware implementation of this software may be carried out either by using conventional peripherals such as LED, <<Buzzer>>, LCD screen or others, or, possibly associated, with new peripherals integrated on the HWP (battery charging circuit) to that end or at the terminal(s) with which the product is set up.\",\n \"For example, the battery level of the product wherein the <<Smart Module>> is integrated may appear at the terminal and not on the product.\",\n \"Power Manager (<<Power Manager>>): This software enables to use the Smart Module as power supply controller.\",\n \"More precisely, this application software enables to control several power supply zones within the same product.\",\n \"The purpose is to control by a software several regulators or transistors which enable to supply or not certain electric zones of the <<Smart Module>> properly speaking and also of the product wherein it is implemented.\",\n \"These regulators and transistors are positioned on the hardware patch.\",\n \"It should be noted that the battery charge management is not taken into account at the <<Power Manager>> software, but at the HWP (battery charge circuit).\",\n \"Management of Inputs and Outputs (<<IO Manager>>): It is a layer which manages all the inputs/outputs between the product and the <<Smart module>>.\",\n \"They are for example of the serial link type (synchronous or asynchronous) or parallel, analogue or digital.\",\n \"On FIGS.\",\n \"2 and 3, a pen 1 is seen in transparency.\",\n \"Inside a box 10 of plastic material, an optical sensor 2 is arranged towards an end in relation to an optical channel 11 of hard material ahead of said pen 1.A communication module of Smart Module type is implemented in the form of a printed circuit 8 including an on/off switching means 3, push-button, a radio-electric component 4 according to the <<Bluetooth®>> protocol, a microcontroller 5 including read-only memory and random-access memory, whereas the memory may be one of the following types: ROM, EPROM, EEPROM, RAM, Flash, SRAM and an electric power supply source 6 in the form of two single-use batteries or of rechargeable batteries.\",\n \"The optical sensor 2, the printed circuit 8, the power supply 6 are held in place by attachment means of the box 10.The pen as claimed is an application of the interface module claimed.\",\n \"The Smart Module used, integrates a two-way radio component and a base band integrated circuit, according to the “Bluetooth®>> standard, as well as a microcontroller enabling to carry on-board, in whole or in part, the <<Bluetooth®>> protocol stack as well as dedicated software applications.\",\n \"An additional microcontroller may be used for the listening phase, corresponding to the measurement of the successive lengths of a barcode.\",\n \"The decoding is performed either on the external microcontroller or on the microcontroller on-board the <<Bluetooth®>> chip.\",\n \"On FIG.\",\n \"4, the previous pen is seen according to a plane perpendicular to the previous one enabling to observe on the printed circuit 8 a light-emitting diode (LED) 9 and an aerial 7 for the radio electric module 4.The aerial 7 is here an aerial of <<wire structure>> type printed on the printed circuit, but it might also be an aerial of <<patch>> type or any other type of radiating element placed inside the pen.\",\n \"The two-colour LED 9 enables to inform the user on the operating condition of the pen.\",\n \"This LED 9 may be replaced with any other type of alarm (sound generator, buzzer for example).\",\n \"The optical sensor arranged at one end of the reader implements reflection optics with a light-emitting diode, for example of 700 nm wavelength, and a photodiode.\",\n \"An aspheric forked lens or, in another embodiment not represented, a system of optical fibres, is arranged at the front of the light-emitting diode and of the photodiode in order to obtain a single focus point at the front of the pen.\",\n \"The output signal of the optical sensor is a current generated in the photodiode and which is amplified by a transistor or an operational amplifier and which is converted into a binary voltage which may be processed by the microcontroller 5.A black surface absorbs the light emitted by the sensor and, in such a case, the output current is weak, the output is at a logic level 1.Conversely, outside a black surface, the light is reflected and the output voltage is at a logic level 0.The reader includes a box 10 and the optical sensor is held in the box at constant distance from the end of the pen, an optical channel 11 being realised between said end and the sensor.\",\n \"Preferably, the optical channel is a light guide (optical fibre for example) or a lens (spherical tip) made of <<hard>> material.\",\n \"The pen type reader of the invention is simple to make and includes preferably a box made of moulded plastic material and implements a reduced number of parts.\",\n \"The electronic circuit of the reader includes a programme in a read-only memory intended for decoding barcodes of one of the following types: EAN, UPC, codes 128, code 39 and possibly specific barcodes.\",\n \"Preferably, the specific barcodes comprise error correction means.\",\n \"Preferably, the specific barcodes are encoded in order to comprise redundancies enabling, during decoding, to correct errors made when reading said barcode.\",\n \"These corporate barcodes are encoded according to the principle of error correction codes used conventionally in the field of telecommunications.\",\n \"Contrary to the state of the art, the pen claimed may read new types of barcodes on display screens, for example of television sets, computers or electronic assistants as well as screens of mobile telephones or any other type of screen using the plasma, TFT, liquid crystal (LCD) technology or by scanning (VGA, XVGA .\",\n \".\",\n \".\",\n \").\",\n \"The barcodes used in such a case are very different from those of the state of the art: these are not codes having any <<spatial extension>> but rather a <<time extension>>.\",\n \"More precisely, the barcodes claimed in the invention correspond to a succession of dark and clear surfaces on the display screen.\",\n \"As a traditional barcode, the information is encoded in the successive durations of the bars and spaces of the code.\",\n \"The user sees this code as a quick flashing of the screen.\",\n \"As this code is decoded by the pen claimed, the user holds said pen in contact with the screen; whereas no displacement is performed on the screen.\",\n \"This characteristic of the pen is a feature of the decoder (software module on-board the pen) and of the analogue circuit carrying out the conversion of the analogue signal from the photodiode, into a digital signal.\",\n \"The main difficulty of this decoding type lies on the one hand in the low contrast between a <<white>> screen and a <<black>> screen.\",\n \"On the other hand, in scanning-type systems, a refreshing frequency is used and complicates the decoding very much.\",\n \"This characteristic being particular mainly to the decoding software, it is independent of the reading technology used (CCD, contact reading, CMOS sensor).\",\n \"This example of reader in the form of a simplified reading pen which is stand-alone with low cost enables implementation of the method in everyday life and more especially in relation to computer means of the hypertext type (hypermedia) on micro-computer, personal assistant, mobile telephone, access point or others.\",\n \"However, any type of reader including the functionalities specified may be implemented with the method of the invention.\",\n \"The element common to the various reading technologies implemented is the interface module or Smart Module.\",\n \"On FIG.\",\n \"4 is represented a block diagram of the pen.\",\n \"Said diagram enables among other things to integrate the on-board software and enables therefore to use the component Bluetooth, on the one hand to send and receive data by radio and on the other hand, to perform the decoding of the barcode, the management of the man/machine interface as well as the management of the power consumption and of the battery.\",\n \"This enables to reduce the complexity (size of memory, calculation power) of the external microcontroller and hence to reduce the hardware cost of the product.\",\n \"The original architecture (software and hardware) of the Smart Module, claimed in the invention, enables therefore to reduce very significantly the cost of a wireless reading pen and may therefore accelerate the implementation of the method in everyday life and more particularly in relation to computer means of the hypertext type (hypermedia) In a particular embodiment of the reader, for example the pen described, the push-button is arranged so that the user positions necessarily the reader in a given orientation with respect to the barcode.\",\n \"The push-button is thus arranged so that the pickup element of the user holds the reader in a given position so that he may depress the button and launch the reading cycle.\",\n \"This given orientation is such that the reader is substantially vertical, i.e.\",\n \"substantially perpendicular with respect to the support of the barcode, when reading.\",\n \"In case when the user should scan the barcode manually, the push-button is also arranged to facilitate said scanning and, for example, so that the pickup element of the user works according to scanning habits used when writing manually.\",\n \"On FIG.\",\n \"5, one has represented a pen 1 when reading a barcode 14 on an object 13.In the close environment which may be reached by radio-electric waves 18 of the module 4 several pieces of computer equipment are arranged which may communicate by electromagnetic waves according to the <<Bluetooth>>® protocol with said pen 1.These pieces of equipment are a micro-computer 15, a mobile telephone 16, with hypertext functionalities, a personal assistant 17 and one or several access points to a local network.\",\n \"Initialisation and Configuration Phase A first initialisation phase is provided for the configuration of the pen.\",\n \"The purpose of this phase is to specify which peripherals are <<twinned>> with the reader.\",\n \"This initialisation method is managed at the processor on-board the Smart Module, at the software Smart Module Manager.\",\n \"It comprises 4 steps: 1—Downloading the customer software on one of the user terminals (micro-computer for example).\",\n \"This software is described schematically on FIG.\",\n \"9.2—The user must then select the set of terminals with which he wants to <<twin>> the pen.\",\n \"Two means are possible to fill in this list: or, the user performs, from the terminal, an <<inquiry>> and selects the terminals found by the inquiry.\",\n \"This requires that the user configures all the terminals in listening mode (<<discoverable>>).\",\n \"Or the user fills <<in long hand>> the list of Bluetooth addresses (48-bit MAC address).\",\n \"Once the list of the terminals is formed, the user must then fill in a field containing the identifier number of the pen (48-bit address of the pen or any other identifier type of the pen).\",\n \"The customer software then prompts the user to switch the pen into listening mode.\",\n \"3—Several means are offered to the user to switch the pen into listening mode (<<discoverable mode>>).\",\n \"The first corresponds to reading a specific barcode (corporate code) and the second method corresponds to the position of the pen in contact with a <<white>> paper for a duration of the order of 1 to 5s approximately.\",\n \"These listening parameters of the pen may evolve and this description is not exhaustive.\",\n \"In parallel, the customer software attempts a connection towards the pen and sends the information specific to the configuration: list of addresses of the peripherals or twinned terminals, the description of the <<profile>> to be used for each terminal, as well as an order of priority associated with each piece of equipment.\",\n \"The NULL or BATCH terminal may be part of the list of the twinned terminals.\",\n \"The purpose is to configure the reader in asynchronous mode: instead of trying to send, over radio, a decoded barcode towards a remote terminal, the pen stores said barcode in its non-volatile memory.\",\n \"4—The configuration data is then stored in the non-volatile memory of the pen and might be modified at each instant by performing again a configuration phase.\",\n \"Upon completed initialisation of the reader, the user is informed of the success of the initialisation phase via the man/machine interface of the pen and/or of the user terminal.\",\n \"Once this initialisation phase has been conducted, the pen may be used as follows: On FIG.\",\n \"6, only the telephone 17 is functional.\",\n \"The link is then established between the equipment and the pen, the data read by the pen is read in the form of sets of digital or alpha-digital or binary data is transmitted by electromagnetic waves 20 towards the equipment 16 as represented on FIG.\",\n \"3c.\",\n \"The transmission of data is performed in real time, the data being transmitted as it is read and decoded or in quasi-real time, the handshake with the equipment or the pieces of equipment of the environment taking place before the transmission, the chains of data being stored temporally in the pen 1 as said link is being established.\",\n \"Between reading 2 consecutive barcodes, the connection is held to maintain reduced consumption.\",\n \"Management of Low Consumption Modes The power consumption is an essential element in the operation of the pen.\",\n \"The <<Bluetooth®>> protocol has a great intrinsic advantage in terms of consumption with respect to other radio standards like 802.11b.\",\n \"Nevertheless, the <<Bluetooth®>> protocol needs to be used in all its complexity and notably in terms of management of low consumption modes.\",\n \"The <<Bluetooth®>> abstraction stack integrates this efficient management of the <<Bluetooth®>> connection modes.\",\n \"When reading the barcode for the first time, the connection is created.\",\n \"When the barcode is sent, the radio connection switches into <<sniff>> mode with a short <<Sniff>> interval.\",\n \"This enables to keep a limited latency time.\",\n \"After a certain time period (T1), the <<sniff>> period is increased in order to reduce the consumption still further.\",\n \"The shortcoming in such a case is an increase in the lead time for sending a barcode.\",\n \"It is nevertheless not a problem since, in such a case, the duration between 2 successive readings is long and the sending lead time is not fundamental.\",\n \"After a certain time (T2), the reader switches into <<parked mode>> after exchanging the master/slave roles.\",\n \"After a time T3, the radio connection is cut off and the power supply of the Smart Module is turned off.\",\n \"The durations T1, T2, T3 are variable parameters which are adapted to the usage mode of the user and provide statistics of durations between successive readings.\",\n \"For a given user reading barcodes at an average high rate, the values of T1, T2 and T3 will be selected as shorter.\",\n \"Multi-Terminal Connection Management FIG.\",\n \"5 represents a pen in its environment.\",\n \"The pen may be set up with different and quite diverse pieces of equipment.\",\n \"It is one of the significant elements of the “Bluetooth®” standard: interoperability and multi-terminal connectivity.\",\n \"Nevertheless, even if these services are provided by the “Bluetooth®” standard, it is still necessary to manage this connectivity in an intelligent way taking into account the fact that the man/machine interface at the pen is very reduced and that it does not enable therefore to select towards which pieces of equipment the pen must be connected.\",\n \"This intelligent management is a feature of the pen and is integrated in the applicative layer of the Smart Module.\",\n \"During the configuration, the devices with which the pen may be connected are recorded in the non-volatile memory of the pen.\",\n \"If the pen has been configured in asynchronous mode (by using only the NULL terminal in the configuration phase), each decoded barcode will be stored automatically in the non-volatile memory of the pen (FIG.\",\n \"7a).\",\n \"In order to access at a later stage the data recorded in the pen, the user must proceed to an upload or charging phase of the data.\",\n \"This phase is performed by switching the pen into listening mode (by reading a corporate code for example) (FIG.\",\n \"7b) while actuating the charging function on the customer software (micro-computer for example).\",\n \"All the barcodes in memory are then transferred by radio waves towards the user terminal (FIG.\",\n \"7c).\",\n \"In the case of a real-time mode configuration (FIG.\",\n \"6), i.e.\",\n \"the pen has been configured with at least one non-NULL (or BATCH) user equipment, the pen tries to connect with different pieces of equipment in memory in order of priority or by using the equipment twinned last.\",\n \"This handshake takes place only at the beginning of a reading phase.\",\n \"A reading phase being characterised in that between 2 successive readings, the duration is shorter than T3 (duration of inactivity before total extinction of the power supply).\",\n \"Throughout the reading phase, the connection is held in low consumption modes as described in the portion called <<management of low consumption modes>>.\",\n \"If none of the pieces of equipment twinned reacts to the handshake from the pen, said pen is configured automatically (after one or several fruitless attempts) in asynchronous mode during the same reading phase.\",\n \"The heterogeneity of the twinned terminals with a pen is very important.\",\n \"The handshake modes, specified in the <<profiles>> of the <<Bluetooth®>> standard are very different of from one type of terminal to another.\",\n \"The on-board software in the Smart Module of the pen is capable of managing several <<profiles>>.\",\n \"More precisely, the applicative abstraction layer selects automatically the appropriate handshake mode in relation to the piece of equipment with which the pen wants to connect.\",\n \"For example, in the case of an access point, the <<profile>> which is adequate is the <<Lan Access Point Profile>>, for a computer or digital assistant, it is the <<Serial Port Profile>> or <<HID profile>> and for a telephone it will be either the <<DialUp Networking Profile>>, by sending AT commands towards the telephone.\",\n \"In such a case, the customer software is reduced to NULL on the terminal: the hypermedia navigator of the telephone being remote-controlled by the pen via AT commands.\",\n \"Other solutions may be implemented, notably, it is possible to download a customer software on the SIM card of the mobile or in the processor memory of the telephone.\",\n \"Battery Management The battery of the pen is rechargeable via an electronic circuit, integrated in the Smart Module.\",\n \"Several voltage regulators, voltage controlled, are used on the Smart Module to control by a software, the power supply of different zones independent of the pen.\",\n \"For example, when the user releases the action button, the zone corresponding to the reading head is not power supplied.\",\n \"Conversely, the power supply of the radio module remains as long as the on-board software requests so.\",\n \"This enables finer management of the consumption of the pen while separating correctly by functionalities the different electronic circuits forming the pen.\",\n \"Man/Machine Interface Management This interface is managed by the software in the Smart Module of the pen.\",\n \"On the pen there are light-emitting diodes or a sound generator.\",\n \"This interface enables to signal the user a good reading or a bad reading, a successful radio send or not.\",\n \"It also enables to inform the user when the battery level is too low.\",\n \"A portion of this man/machine interface can be delocalised on the terminal when said terminal is provided with a richer interface (micro-computer or mobile telephone for example) In its application, the invention is structured around four components: the barcode reader, the customer equipment, the routing server, the document server, possibly reached by a relay server (proxy).\",\n \"Anyone who wishes to access the information and services associated with a resource uses the barcode reader to scan the barcode referring to this resource.\",\n \"The reader then decodes the barcode to generate a resource identifier.\",\n \"Said identifier is sent with the own identifier of the reader towards the customer equipment, through a local connection.\",\n \"The customer equipment processes the identifiers received to determine the address of an appropriate document server.\",\n \"This server has the hypermedia documents exhibiting the information and services relative to the resource designated with the reader.\",\n \"The customer equipment may, to facilitate its address research, obtain additional information from the routing server.\",\n \"The identifiers from the reader are then relayed through a remote connection towards the document server, whereof the address has just been determined by the customer equipment.\",\n \"Additional information whereof the user has authorised the broadcast may at this stage be transmitted over by the same channel.\",\n \"Possibly, the remote connection towards the document server goes through a relay server or proxy, which is capable of storing the request and/or of processing it for statistical purposes or others, before transmitting it integrally to the document server.\",\n \"Finally, the document server processes the data which has been transmitted thereto in order to send back to the customer equipment the hypermedia document which will enable the user to access the information and services requested.\",\n \"All these steps take advantage of the latest technological breakthroughs in the field of telecommunications to offer real-time or quasi real-time service: there are only a few seconds between the scanning of the barcode and the presentation of the associated hypermedia document.\",\n \"The barcode reader is a piece of hardware equipment capable of decoding a barcode into electric signals then into digital and/or alpha-digital data.\",\n \"Such data forms an identifier of the resource referred to by the barcode.\",\n \"This resource identifier and another identifier personal to the reader are both transmitted towards the customer equipment via a local connection.\",\n \"The reader is intended for general public and consequently should be cheap enough.\",\n \"It may conversely be of modest quality in comparison with industrial readers since its use is vastly less intensive.\",\n \"it can be broken down into four main portions: an optical transceiver, a power supply, a local transmitter, a box.\",\n \"The local connection is a short-range communication link between two pieces of equipment.\",\n \"The distance between the pieces of equipment does not exceed a few ten metres.\",\n \"Comprised as an electronic device, the local connection locale designates a short-range transceiver controlled via a communication protocol.\",\n \"The customer equipment is the central element of the system.\",\n \"The customer equipment is a piece of user equipment implementing the customer software.\",\n \"The user equipment is a piece of hardware equipment possessed by the user.\",\n \"It may have various characteristics, mobile or not, providing the following elements are available: a local connection, a remote connection, an operating system with a documented programming interface, a hypermedia navigator.\",\n \"Concretely, a piece of mobile equipment may be a computer or a digital assistant connected to the Internet, a WAP mobile telephone offering programming possibilities, a GPRS, UMTS terminal or any other piece of equipment meeting the conditions detailed previously.\",\n \"The remote connection is world-wide communication link between two pieces of equipment.\",\n \"It almost always implements global networks, whereof the Internet is the best representative.\",\n \"In this context, it may be contemplated to have all the requests transit via a set relay server (proxy), so that said relay updates the global information regarding the set of requests.\",\n \"In all cases, the requests are at least transmitted integrally.\",\n \"Comprised as an electronic device, the remote connection designates a network transceiver, possibly controlled via communication protocols.\",\n \"The customer software is a piece of software running on any type of user equipment to fulfil the following functions: receive the identifiers transmitted by the barcode reader via the local connection, provide the user with the hypermedia document for accessing the information and services relative to the resource designated by the reader.\",\n \"This piece of software is called customer software since it behaves as a customer for the routing server on the one hand and for the document server whereof it must determine the address on the other hand.\",\n \"It is realised as an application programme, for example using an object-oriented programming language such as C++ or Java.\",\n \"It controls the user equipment via the programming interface specific to such equipment.\",\n \"The installation of this piece of software on a piece of user equipment may require a programme to be downloaded, the insertion of an electronic board whereon it is implanted (chop card for mobile telephone, extension card for portable computer or digital assistant) or any other electronic or computerised method.\",\n \"The customer software may be broken down into three modules: a local receiver, a router, a navigator manager.\",\n \"The local receiver is a programme whereof the role consists in listening further for the local connection.\",\n \"It reacts to any message from the barcode reader while activating the router and while transmitting to said reader the identifiers received from the reader.\",\n \"This programme is running as long as the system is actuated.\",\n \"Conversely, when no message from the reader is being processed, it is the single portion of the customer software in operation, which reduces the consumption of the resources of the user equipment.\",\n \"The router is a programme receiving from the local receiver the resource identifier and the reader identifier.\",\n \"Its role consists in applying to these identifiers the address resolution algorithm to attempt to determine the WEB address of a document server appropriate to the resource referred.\",\n \"If such an address is obtained, the router adds according to a set syntax both previous identifiers, as well as certain information which the user may have chosen to provide on its equipment (for example certain preferences or restrictions).\",\n \"The address thus completed locates without any ambiguity a hypermedia document exhibiting information and services relative to the resource identified via the reader.\",\n \"The router then actuates the navigator manager and transmits to said manager the address of the document.\",\n \"The address resolution algorithm is applied to the resource and reader identifiers.\",\n \"This algorithm processes first of all the reader identifier.\",\n \"If the identifier is known, i.e.\",\n \"if it listed on a routing table (correspondence) stored on the user equipment and managed by the router, then the programme extracts from the same table the routing information regarding the reader.\",\n \"The routing table may be realised with diverse data structures; it forms a very simplified system for database management.\",\n \"The routing information may contain: a history of the previous requests with their result, whereof one will be able to extract the address of a document server; a single address of document server, for example in case when the reader is exclusively affiliated to such server or a relay server address, through which all the requests pertaining to a document server should transit; any other information peculiar to the reader and useful to the address resolution.\",\n \"In a second stage, the address resolution algorithm processes the resource identifier and the routing information extracted during the first step.\",\n \"In some cases, this data is sufficient to generate the address of the document server associated with the resource referred.\",\n \"For example, it may be agreed that an identifier exhibiting certain similarities with another identifier processed during a previous request must be associated with the address which resulted from this request.\",\n \"If is not possible to conclude with this single data, the router transmits to the routing server the reader identifier and the resource reader, via the remote connection.\",\n \"This server returns to the router the address of the document server which it is waiting for, as well as possibly the address of a relay server (proxy) associated, and these addresses are stored in the routing table.\",\n \"The address resolution algorithm has therefore enabled to determine the address of the document server with at most a remote access to the routing server.\",\n \"It should be noted however that it may be necessary to refresh periodically the routing information, via additional requests transmitted to the routing server during possible remote access.\",\n \"In case when the reader identifier is not known from the router, the behaviour of the latter depends on the operating mode wherein the user has placed its customer equipment.\",\n \"In current mode, the resolution procedure is aborted and the message from the reader is therefore ignored.\",\n \"This normal situation may for example crop up if a message radio-transmitted by a user's reader is received by the customer equipment of another user, who is not interested at all in his neighbour's intentions.\",\n \"If, conversely, the user has placed its customer equipment in receiving mode of a new barcode reader, then the new reader identifier is transmitted towards the routing server via the remote connection, so that this server sends back to the router the routing information peculiar to the new reader.\",\n \"The receiving mode enables therefore to record a new reader.\",\n \"Such information is placed in a new entry to the routing table, and the processing of the request by the customer software is completed.\",\n \"The navigator manager is a programme whereof the role consists in causing the hypermedia navigator available on the user equipment to provide the hypermedia document located by the address received from the router.\",\n \"The user is then free to navigate at will from the document which is presented to him.\",\n \"The navigator manager only controls the display of a document to the usual navigator of the user, who remains perfectly free to set it up at his discretion.\",\n \"To illustrate the mechanism implemented, one may consider the example of a WEB navigator to whom the navigator manager will order the creation of a new window to display an HTML document.\",\n \"The address of this document is provided to the navigator in the form of a URL complying with the HTTP protocol.\",\n \"Another example consists in reproducing on a mobile telephone connected to the Internet a sound document, located by an address complying with the WAP protocol.\",\n \"The routing server is the auxiliary information source of the <<Baracoda®>> system (denomination adopted for the set of components, methods and services linked with the invention).\",\n \"The routing server is a piece of software running on one or several computers fitted with a remote connection to fulfil the following functions: manage and update the routing information peculiar to each barcode reader; manage and update the information enabling to associate the WEB address of a relay server with the WEB address of a document server; manage and update the information enabling to associate the WEB address of a document server to a resource identifier accompanied by a reader identifier; manage and update the code of the different programmes forming the customer software; listen further to the remote connection in order to react to the requests of a customer equipment by transmitting to said customer the information that he requests.\",\n \"It behaves therefore as a server for each piece of customer equipment, whereas the number of customers may be quite high.\",\n \"It responds to the different requests of the router, but also to installation requests from a piece of user equipment that has not been fitted with the customer software.\",\n \"The software routing server may be a single software running on a single computer, or be distributed on several computers.\",\n \"The network organisation of these machines may itself implement diverse architectures: a hierarchical architecture, according to which the machines situated at the lowest nodes hold local information and refer to higher rank machines to obtain more general information; a neighbouring architecture, each machine being at the same rank as its neighbours and working in co-operation with the latter to access largely distributed information; any other architecture deemed efficient to share the processing load and capable of improving the response time to requests.\",\n \"The communication between the routing server and a piece of customer equipment may employ a corporate protocol specifically designed to that end or a public protocol (HTTP or WAP for example).\",\n \"Different protocols may be selected according to the nature of the customer equipment (computer, digital assistant, mobile telephone, etc.).\",\n \"The document server is the last element of the ‘<Baracoda®>> system.\",\n \"The document server is a piece of software running on one or several computers fitted with a remote connection to fulfil the following functions: manage and update a large number of hypermedia documents showing the information and services relative to certain resources; listen further to the remote connection in order to react to the requests of a hypermedia navigator by transmitting to said navigator the document that he requests.\",\n \"The routing tables stored on the customer equipment or on the routing server comprise not only the addresses of document servers but also the addresses of relay servers associated with the document servers.\",\n \"The software component called “router” of the “customer software” is therefore capable of providing the component called “navigator manager” with an address based on that of the relay server, so that the latter filters all the requests send to a given document server.\",\n \"The relay server only re-transmits the request to the document server which is the final assignee thereof.\",\n \"But it may in passing memorise that a request has been made and perform statistical processing of all the requests thus relayed.\",\n \"This additional service offered to certain document servers may constitute an added value for anyone whose wishes to know the look-up frequency based upon the barcodes, the total volume of requests, their time distribution, etc.\",\n \"The document server is the property of a third party wishing to provide the documents relative to certain resources.\",\n \"It is implemented and managed to the discretion of the owner.\",\n \"For example it may be the server of an industrial company publishing its catalogue of products on the Internet.\",\n \"However, it must in order to be integrated into the <<Baracoda®>> system, be capable of interpreting the address transmitted by the navigator manager via the navigator.\",\n \"To guarantee such compatibility at the lowest possible cost, the address considered is in most cases in compliance with a public protocol.\",\n \"It may be for example URL, HTTP or WAP.\",\n \"In such a case, the upgrade of an existing WEB server only requires minor adaptation, for example the design of a single page of dynamic redirection.\",\n \"The application of the radiocommunication module is not limited to the method for reading barcodes and it is possible to implement the module in other applications.\",\n \"For example, the <<Bluetooth®>>—GPRS router is an electronic apparatus composed of a <<SmartModule>> and of a modem GSM, GPRS, EDGE or UMTS.\",\n \"Said apparatus carries out the conversion of protocol between the <<Bluetooth®>> communication protocol and the GPRS, EDGE or UMTS protocol.\",\n \"It enables for example <<Bluetooth®>> products to access a remote network (Internet network for example).\",\n \"The applications of such apparatus are numerous in the field of house computerised appliances and the field of Internet access (ISP) for a residential user or a small company.\",\n \"The apparatus consists of a radiocommunication module and of a GSM/GPRS/EDGE/UMTS radio with management of the physical layer of the GSM/GPRS/EDGE/UMTS protocol and of the protocol stack considered as well as of a specific software managing the quality of service and the sharing of the pass-band between the different users.\",\n \"Up to 7 products fitted with the apparatus may use the services of the GPRS/UMTS simultaneously, via the same <<Bluetooth®>>—GPRS router.\",\n \"More than 7 pieces of equipment (up to 255) may access these services via the same router, but not simultaneously.\",\n \"These pieces of equipment may use a token protocol (developed on the processor of the router) in order to send or receive data regularly using the GPRS/UMTS connection.\",\n \"Thanks to the apparatus which integrates the necessary hardware and software, the user does not need to manage the complexity of the routing software due to the necessity of sharing the pass-band in the ascending direction (from the <<Bluetooth®>>—GPRS access point towards the GPRS base station).\",\n \"The <<Bluetooth®>>—GPRS router is also used as a <<Hub>> within the framework of the construction of a local <<Bluetooth®>> network.\",\n \"One may also implement such router in house computerised appliances.\",\n \"Refrigerators, alarms, radiators, lighting units, on/off switches, thermometers and others sensors (water, gas, electricity meters), television set, VCRs may be connected to a remote network via a <<Bluetooth®>>—GPRS access point which is this <<Bluetooth®>>—GPRS router.\",\n \"In such an application of the router it is for example possible to activate the heating system of one's country house via its mobile telephone, remotely.\",\n \"The router may also be applied for accessing Internet at home with PCs or PDAs (portable electronic assistant) or inside a car.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10474185"}}},{"rowIdx":787,"cells":{"text":{"kind":"list like","value":[["Method and divece for outputting data relating to information of a motor vehicle","In a method for outputting data relating to user selectable information regarding a motor vehicle, a plurality of user selectable options relating to the information are represented, and data relating to the selected option is output.","At least one context-dependent profile is defined, and a set of options determined by the profile is generated.","Data are output as a function of the profiles and options."],["1-15.","(canceled) 16.A method for outputting to a vehicle user, data relating to information concerning a motor vehicle, said method comprising: defining at least one context-dependent profile; generating user selectable options for at least one item of information, which options are characterized by the at least one context-dependent profile; and outputting data relating to a selected option, said data being selected from an entire data set as a function of the at least one profile.","17.The method as claimed in claim 16, wherein an operator control function which corresponds to the data which are output is executed as a function of the profiles and options.","18.The method as claimed in claim 16, wherein a particular profile is provided for a predefinable user group.","19.The method as claimed in claim 16, wherein a particular profile is provided for a predefinable vehicle environment.","20.The method as claimed in claim 16, wherein a particular profile is provided based on the type of vehicle.","21.The method as claimed in claim 16, wherein the options are presented and/or the data are output as a function of current vehicle parameters.","22.The method as claimed in claim 16, further comprising: performing a plausibility check for a control operation that is carried out by the vehicle user; and presenting the options or outputting the data are output as a function of a result of said check.","23.The method as claimed in claim 16, further comprising: storing preceding vehicle variables and/or preceding control operations; and presenting options or outputting the data, as a function of stored values.","24.The method as claimed in claim 16, further comprising: processing the data further; and feeding the data to other systems in the motor vehicle.","25.The method as claimed in claim 16, wherein at least part of a motor vehicle operating manual is presented as information.","26.A device for outputting to a vehicle user, data relating to information concerning a motor vehicle, said device comprising: a processing unit which generates, by reference to at least one defined context-dependent profile, a plurality of options which are associated with the at least one profile; interface means for representing the options generated by the processing unit; means for a user selecting at least one option from among the options generated by the processing unit and represented by the interface means; and means for outputting data relating to a selected option, as a function of the at least one profile.","27.The device as claimed in claim 26, further comprising at least one display unit for outputting data as a function of a data set which characterizes it.","28.The device as claimed in claim 26, wherein the selection means comprises a voice input.","29.The device as claimed in claim 26, wherein the selection means comprises a manual operator control element.","30.The device as claimed in claim 26, wherein means are provided for audibly outputting options and/or data.","31.A method for selectively outputting vehicle related information to a vehicle user, comprising: providing an overall data set that includes data relating to said vehicle, said data being accessible according to a hierarchical classification structure, including a plurality of items of information, each of said items of information having associated therewith selected data from said overall data set, said selected data being selectively displayable according to a plurality of selectable options associated with said information; defining a plurality of context dependent profiles whose content is based on parameters characterizing at least one of said vehicle, its operation and said vehicle user; in response to designation of an item of information, generating a set of selectable options from among said plurality of selectable options, based on a particular context dependent profile; and outputting data relating to a selected option, as a function of the particular option."],["<SOH> BACKGROUND AND SUMMARY OF THE INVENTION <EOH>The invention relates to a method and apparatus for outputting data providing information regarding a motor vehicle to a user of the motor vehicle.","International Patent Document WO 98/34812 discloses a multimedia information and control system for a motor vehicle, in which the user selects the information of interest to him from a defined global set.","Various options relating to the information are then offered, from which he can select at least one option and cause the respective data to be output.","Furthermore, an operator control function which corresponds to the data that are output can be executed.","One object of the invention is to provide an improved method and apparatus of the generic type in which operating convenience for the user is increased and operating safety and traffic safety are improved.","This and other objects and advantages are achieved by the method and apparatus, according to the invention, in which the type and number of the options generated with respect to the information is determined and the respective data are output, as a function of at least one context-dependent profile, thus providing the user with fast and simple access to data which are more wide ranging in that they describe the individual information more closely.","As used herein, the word “information” designates a generic classification or heading under which various data can be grouped, for example “windshield wiper”.","The data relating to particular information comprises assigned details which are stored in a memory and which can be output in different ways for the user: in the case of “windshield wiper”, for example, an introduction video for operator control or a brief audio explanation of the interval levels.","The context-dependent profile relates to the characterization of at least one secondary factor in the interaction between the vehicle, the vehicle user and the environment (for example the user's age).","By reference to the context dependent profile, the options which are associated with the “information” (e.g., windshield wipers) are conditioned in such a way that from a total set of options, only those options which are assigned to the context-dependent profile are generated.","Furthermore, by using the context-dependent profile it is also possible to output data automatically, without the user's previously having selected an option.","By means of the context-dependent profile the number of generated options from which the user can make a selection and the outputting of the data are determined in such a manner that a clearly organized and user-compatible representation is made possible.","Thus, when an item of information relating to the motor vehicle is requested, a subset of the entire set of data relating to this information is selected as a function of the profiles.","(The subset can of course also correspond to the entire data set.)","The selected data are output in a way which is particularly data-optimized for the user.","As a result, the sequence of selecting data and outputting data is shortened, depending on the context.","The invention also makes a contribution both to convenience and to safety.","For example, when a vehicle display lights up, the user is automatically offered an explanation of this vehicle display as a function of the profile, by reason of appropriate data being output.","The user is thus immediately informed, without having to think about the explanation or even perform awkward control operations.","Likewise it possible to provide the user with selective proposals as a function of the profiles and to instruct him about vehicle functions which are suitable to use in a specific situation.","It is advantageous if an operator control function whose use is proposed with the data which is output, is triggered as a function of the profiles and options.","As a result, the operating convenience is increased further, because the user does not need to search for the triggering means which are assigned to this operator control function by virtue of the fact that the proposed operator control function is triggered in an easy way which is optimized as much as possible with respect to time.","Such direct triggering of the operator control function, without previous interrogation of the user makes it possible to trigger the operator control function much more reliably in particular in driving situations for which timing is critical.","Alternatively, direct triggering of the operator control function may be provided.","In this case, as a function of the profiles, a corresponding option is defined, said option being selected and then bringing about indirect triggering of the operator control function.","An example of direct or indirect triggering occurs if the driver selects the reverse gear on his vehicle, and as a result, either an electrically movable rear roller blind is retracted (direct triggering) or the retraction of the rear roller blind is proposed as an option (indirect triggering).","One advantageous embodiment of the invention selects, for a particular user, a context-dependent profile based on the assignment of that user to a defined user group.","As a result, convenience is increased further in that a specific, group related information requirement is fulfilled.","For example, the assignment of the user to a user group takes into account whether he or she is already familiar with the vehicle or the make of vehicle.","For example, a first user group may comprise users for which the vehicle or the make of vehicle is new, and a second user group may comprise experienced users.","If, for example, a user is assigned to the first user group, a specific context dependent profile is defined, bringing about an additional, explanatory outputting of data, for example relating to operator control elements.","Alternatively or additionally, a brief explanation on the vehicle and its operator control elements is possible when the user is assigned to the first user group.","The described assignment to user groups is carried out, for example, during visits for servicing of the vehicle.","Thus, when the vehicle is handed back the “new user” profile can then be defined.","On one of the subsequent visits to the workshop it is then possible to define the “experienced users” group.","A further possibility of a predefinable user group is to select an “interested user” profile which, for example with respect to predefinable vehicle reactions, supplies data in the form of explanations or technical explanations on vehicle functions.","By assigning the user to a user group for the disabled it is possible to output data as a function of the profile in order to support the user in specific situations.","For example, given an assignment of the user to a “restricted hearing” user group, as a function of the profiles, supplementary visual data are output if according to standard practice an operator control element provides audible feedback in the vehicle.","A user can also be assigned to more than one user group, so that more than one profile is assigned for him, preferably with different priorities assigned to different profiles.","If a signal for representing options and/or for outputting data (below for the sake of simplification we only refer to the outputting of options or data) from two or more profiles is present, the options or data are output in correct order and as a function of the priority of the profile.","Here, for example, the options or data with the highest priority are output immediately and additionally an option which the user can select in order to cause the further options or data to be output is illustrated.","It is also possible to define separate profiles for a plurality of users, for example for a user who drives the vehicle, a user who is a front seat passenger, or a user who is sitting in the rear of the vehicle.","In a profile which is defined for the driver of the vehicle, functions relating, for example, to driving safety are provided, while in a profile defined for the rear-seat passengers, audio and multimedia functions are provided, for example.","In this manner, the operating reliability and thus the traffic safety are increased by the context-dependent profile for the driver of the vehicle.","Similarly, for a rear-seat passenger the operating convenience is increased, and comprehensive information about the operator control functions which are available is provided.","In addition it is also thus possible to define further profiles, for example it is possible to provide a specific profile for a service technician with a representation of options and/or outputting of data on the diagnostics of the vehicle.","A further advantageous embodiment of the invention selects a context-dependent profile for a predefinable vehicle environment.","Here, a user is informed about changes which have occurred in the course of his journey by presenting options and/or data which permit the user to prepare for changed conditions on his route, as a function of the profiles.","For example, it is possible for him to obtain information about operator control functions which are suitable for the new conditions and to trigger these functions.","Thus, in the case of a road in poor condition which is stored as an attribute in the digital road map of the vehicle, it is possible to present options as a function of the profiles and/or to output data relating to the operator control elements of a ride level controller (which sets the ground clearance of the motor vehicle).","Furthermore, it is possible, for example, to define a country-specific profile.","When it is detected that a national border has been crossed, information about country-specific traffic regulations are provided as a function of the profiles.","A conventional device for locating the position of the vehicle is used, for example, to identify the border crossing.","In a further refinement of the invention, context dependent profiles are defined with respect to the type of vehicle.","As a result, a user is informed about changes which occur when the type of vehicle which he uses at a particular time changes.","If the user changes, for example, from his previous vehicle model of his make of vehicle to a newer model, only options or data which relate to operator control elements which have changed or are new in comparison to his previously used vehicle are issued to him.","This applies in an analogous way, for example, to retrofits or software updates of vehicles.","In each case the differences between the current vehicle and the vehicle which is known to the user is determined first, and only those options or data which are associated with this difference for the user are output to the user as a function of the profiles.","As a result, by virtue of the profiles, the user is informed about the possibilities which the currently used vehicle provides.","These context-dependent profiles are preferably defined for a user individually according to a type of vehicle.","It is possible, for example, for a “first user” profile to be present which explains innovations only a single time, while a “second user” profile explains these changes within a specific time period whenever a journey is begun.","It is very advantageous to define context dependent profiles which output options and/or data as a function of at least one current vehicle variable, such as for example, currently measured sensor data or vehicle system states such as the heating level of the seat heating system or the range of the vehicle with the currently available quantity of fuel which is determined by the trip computer.","As a result, a vehicle user can be informed particularly quickly about specific situations because, data corresponding to the situation indicated by the vehicle variables is issued to him, as a function of the profiles.","For example, options or data relating to a TCS (Traction Control System) can be output if the wheels spin during the process of driving off.","It is also possible to output, as a function of the profiles, instructions on “cutting free” (by the user briefly switching off the TCS during the process of driving off), by virtue of the fact that the operator control element for switching off the TCS is pointed out and/or direct triggering—i.e.","deactivation—with or without previous interrogation of the user is brought about.","In the case of a flat tire or an accident it is possible, for example, to issue options or data on the correct behavior in emergency situations and/or on the position of the warning triangle in the vehicle.","The vehicle variables can also be transmitted from outside the vehicle, for example in wireless fashion by XFCD (“Extended Floating Car Data”).","It is thus possible to transmit into the vehicle sensor data about weather conditions on the route ahead.","The user is then issued with options or data as a function of the profiles such that the user can set an air conditioning system in his vehicle in a way which is advantageous for these conditions.","A further advantageous embodiment of the invention provides for a plausibility check for a control operation which is carried out by the user, with the options and/or the data being output as a function of the result of the plausibility check.","This increases the operator control reliability and increases the convenience because a user is offered information about an operator control function or its operator control element as a function of the profile if he makes an operator control error or if an algorithm detects an operator control problem.","As a result, it is possible, for example when there is multiple use of the operator control elements for main beam and/or indicator lights within a short period of time while it is raining at the same time, to offer and/or issue data on the operator control of the windshield wiper, in particular on how to switch it on, as options for selection.","In a further configuration of the invention, vehicle variables and/or operator control operations are stored in a memory, which is initially interrogated before options are represented and/or data is output as a function of the profiles.","If values relating to preceding vehicle variables or operator control operations are stored in the memory, these values are checked to determine whether they are to be taken into account in the representation of the current options and/or in the outputting of the current data.","This further increases operating convenience because a rapid selection of options and/or data is matched individually to the user.","As it were, a “memory” is provided by virtue of the fact that conclusions can be drawn relating to a current operator control operation, for example from analyzing preceding operator control operations.","In the case of operator control steps which the user usually makes, it is thus possible to suppress a representation of information and outputting of data.","The classification of operator control problems or the categorization of current vehicle variables is also improved by virtue of the fact that, for example, operator control steps which are specific to a user are detected and no longer classified as an operator control problem and explanatory data is output.","Such a recognition can be achieved, for example, by using the methods of “artificial intelligence” (such as a neural network).","By checking of values stored in the memory it is also possible to make modifications to the currently defined, context-dependent profile itself and/or retrieve and subsequently define a different context dependent profile.","It is advantageous if at least part of an operating manual for the motor vehicle is represented as information relating to the motor vehicle.","A method suitable for this purpose is described, for example, in the German Patent Application DE 199 59 755.3-21 whose content is included herein by reference.","In this method, a current equipment variant of the motor vehicle is determined in a preceding step and a second database is generated from a first database as a function of this determined variant, this second database comprising only information on the operator control for the current equipment variant of the motor vehicle.","In this way, a continuously updated information state relating to the motor vehicle and its operator control elements is ensured.","The device according to the invention for processing and outputting data relating to information regarding a motor vehicle or its operator control elements comprises a display for representing options relating to the information (referred to below for short as first output means), a display for the user to select at least one option (referred to below for short as selection means), a display for outputting data relating to the selected option (referred to for short below as second output means) and a processing unit which generates, by reference to at least one defined, context-dependent profile, a number of options that are characterized by the profile and which processing unit outputs data as a function of the profiles and options.","This processing unit may be embodied, for example, as a vehicle computer (a processing unit which is arranged in the vehicle).","However, it can also be implemented by providing additional functions to a unit which is already present in the vehicle—for example a control device.","The first output means and/or the second output means may be, for example, display units of conventional design such as LCD (Liquid Crystal Display), or may be presented on the windshield (Head Up Display).","It is advantageous here to embody the display unit as a function of the set of characteristics of the options and/or data (both of these are referred to below as data set).","For example, a small data set may be output by means of a display which is already present in the vehicle, while a relatively large data set is output by means of a further relatively large display which is mounted on the dashboard or in the center console.","In addition, further possibilities for displaying options and/or data are available.","The outputting of the options and data can also depend on what technical equipment a vehicle has.","For example, when a suitable display fails, it is possible to suppress the outputting of certain data (for example video sequences forming a short introduction to the vehicle for a new user).","Of course, it is also possible to provide a joint display for outputting the options and data.","In order to integrate the first and/or second output means with the selection means, a conventional touch-sensitive display (touch screen) is preferably provided.","Alternatively or additionally, the selection means may be embodied as a voice operated controller, such as is already used today in motor vehicles, permitting the driver to concentrate his visual attention on the events on the road.","Furthermore, the selection means may be embodied as a manual operator control element of conventional design, for example, in the form of a joystick, or as a combined rotary/pushbutton knob which has in each case a different number of latching positions depending on the number of options for the information.","Such an operator control element is also already present in many vehicles (for example for controlling a navigation device), and can be used advantageously.","Further possible operator control systems may be provided.","In order to output the options or the selected data it is possible to provide means of a conventional design, for example audible outputs, such as an audio system, or haptic output (for example a force feedback operator control element, an opposing force being exerted at said operator control element when it is used).","The means which is used for inputting and/or outputting may also be defined in a variable fashion as a function of the profiles, for example a visual output is replaced by an audible output as soon as the vehicle exceeds a defined limiting speed.","Furthermore, the data which are output can be further processed and fed to other systems, in particular assistance systems in a motor vehicle.","Other objects, advantages and novel features of the present invention will become apparent from the following detailed description of the invention when considered in conjunction with the accompanying drawings."],["<SOH> BACKGROUND AND SUMMARY OF THE INVENTION <EOH>The invention relates to a method and apparatus for outputting data providing information regarding a motor vehicle to a user of the motor vehicle.","International Patent Document WO 98/34812 discloses a multimedia information and control system for a motor vehicle, in which the user selects the information of interest to him from a defined global set.","Various options relating to the information are then offered, from which he can select at least one option and cause the respective data to be output.","Furthermore, an operator control function which corresponds to the data that are output can be executed.","One object of the invention is to provide an improved method and apparatus of the generic type in which operating convenience for the user is increased and operating safety and traffic safety are improved.","This and other objects and advantages are achieved by the method and apparatus, according to the invention, in which the type and number of the options generated with respect to the information is determined and the respective data are output, as a function of at least one context-dependent profile, thus providing the user with fast and simple access to data which are more wide ranging in that they describe the individual information more closely.","As used herein, the word “information” designates a generic classification or heading under which various data can be grouped, for example “windshield wiper”.","The data relating to particular information comprises assigned details which are stored in a memory and which can be output in different ways for the user: in the case of “windshield wiper”, for example, an introduction video for operator control or a brief audio explanation of the interval levels.","The context-dependent profile relates to the characterization of at least one secondary factor in the interaction between the vehicle, the vehicle user and the environment (for example the user's age).","By reference to the context dependent profile, the options which are associated with the “information” (e.g., windshield wipers) are conditioned in such a way that from a total set of options, only those options which are assigned to the context-dependent profile are generated.","Furthermore, by using the context-dependent profile it is also possible to output data automatically, without the user's previously having selected an option.","By means of the context-dependent profile the number of generated options from which the user can make a selection and the outputting of the data are determined in such a manner that a clearly organized and user-compatible representation is made possible.","Thus, when an item of information relating to the motor vehicle is requested, a subset of the entire set of data relating to this information is selected as a function of the profiles.","(The subset can of course also correspond to the entire data set.)","The selected data are output in a way which is particularly data-optimized for the user.","As a result, the sequence of selecting data and outputting data is shortened, depending on the context.","The invention also makes a contribution both to convenience and to safety.","For example, when a vehicle display lights up, the user is automatically offered an explanation of this vehicle display as a function of the profile, by reason of appropriate data being output.","The user is thus immediately informed, without having to think about the explanation or even perform awkward control operations.","Likewise it possible to provide the user with selective proposals as a function of the profiles and to instruct him about vehicle functions which are suitable to use in a specific situation.","It is advantageous if an operator control function whose use is proposed with the data which is output, is triggered as a function of the profiles and options.","As a result, the operating convenience is increased further, because the user does not need to search for the triggering means which are assigned to this operator control function by virtue of the fact that the proposed operator control function is triggered in an easy way which is optimized as much as possible with respect to time.","Such direct triggering of the operator control function, without previous interrogation of the user makes it possible to trigger the operator control function much more reliably in particular in driving situations for which timing is critical.","Alternatively, direct triggering of the operator control function may be provided.","In this case, as a function of the profiles, a corresponding option is defined, said option being selected and then bringing about indirect triggering of the operator control function.","An example of direct or indirect triggering occurs if the driver selects the reverse gear on his vehicle, and as a result, either an electrically movable rear roller blind is retracted (direct triggering) or the retraction of the rear roller blind is proposed as an option (indirect triggering).","One advantageous embodiment of the invention selects, for a particular user, a context-dependent profile based on the assignment of that user to a defined user group.","As a result, convenience is increased further in that a specific, group related information requirement is fulfilled.","For example, the assignment of the user to a user group takes into account whether he or she is already familiar with the vehicle or the make of vehicle.","For example, a first user group may comprise users for which the vehicle or the make of vehicle is new, and a second user group may comprise experienced users.","If, for example, a user is assigned to the first user group, a specific context dependent profile is defined, bringing about an additional, explanatory outputting of data, for example relating to operator control elements.","Alternatively or additionally, a brief explanation on the vehicle and its operator control elements is possible when the user is assigned to the first user group.","The described assignment to user groups is carried out, for example, during visits for servicing of the vehicle.","Thus, when the vehicle is handed back the “new user” profile can then be defined.","On one of the subsequent visits to the workshop it is then possible to define the “experienced users” group.","A further possibility of a predefinable user group is to select an “interested user” profile which, for example with respect to predefinable vehicle reactions, supplies data in the form of explanations or technical explanations on vehicle functions.","By assigning the user to a user group for the disabled it is possible to output data as a function of the profile in order to support the user in specific situations.","For example, given an assignment of the user to a “restricted hearing” user group, as a function of the profiles, supplementary visual data are output if according to standard practice an operator control element provides audible feedback in the vehicle.","A user can also be assigned to more than one user group, so that more than one profile is assigned for him, preferably with different priorities assigned to different profiles.","If a signal for representing options and/or for outputting data (below for the sake of simplification we only refer to the outputting of options or data) from two or more profiles is present, the options or data are output in correct order and as a function of the priority of the profile.","Here, for example, the options or data with the highest priority are output immediately and additionally an option which the user can select in order to cause the further options or data to be output is illustrated.","It is also possible to define separate profiles for a plurality of users, for example for a user who drives the vehicle, a user who is a front seat passenger, or a user who is sitting in the rear of the vehicle.","In a profile which is defined for the driver of the vehicle, functions relating, for example, to driving safety are provided, while in a profile defined for the rear-seat passengers, audio and multimedia functions are provided, for example.","In this manner, the operating reliability and thus the traffic safety are increased by the context-dependent profile for the driver of the vehicle.","Similarly, for a rear-seat passenger the operating convenience is increased, and comprehensive information about the operator control functions which are available is provided.","In addition it is also thus possible to define further profiles, for example it is possible to provide a specific profile for a service technician with a representation of options and/or outputting of data on the diagnostics of the vehicle.","A further advantageous embodiment of the invention selects a context-dependent profile for a predefinable vehicle environment.","Here, a user is informed about changes which have occurred in the course of his journey by presenting options and/or data which permit the user to prepare for changed conditions on his route, as a function of the profiles.","For example, it is possible for him to obtain information about operator control functions which are suitable for the new conditions and to trigger these functions.","Thus, in the case of a road in poor condition which is stored as an attribute in the digital road map of the vehicle, it is possible to present options as a function of the profiles and/or to output data relating to the operator control elements of a ride level controller (which sets the ground clearance of the motor vehicle).","Furthermore, it is possible, for example, to define a country-specific profile.","When it is detected that a national border has been crossed, information about country-specific traffic regulations are provided as a function of the profiles.","A conventional device for locating the position of the vehicle is used, for example, to identify the border crossing.","In a further refinement of the invention, context dependent profiles are defined with respect to the type of vehicle.","As a result, a user is informed about changes which occur when the type of vehicle which he uses at a particular time changes.","If the user changes, for example, from his previous vehicle model of his make of vehicle to a newer model, only options or data which relate to operator control elements which have changed or are new in comparison to his previously used vehicle are issued to him.","This applies in an analogous way, for example, to retrofits or software updates of vehicles.","In each case the differences between the current vehicle and the vehicle which is known to the user is determined first, and only those options or data which are associated with this difference for the user are output to the user as a function of the profiles.","As a result, by virtue of the profiles, the user is informed about the possibilities which the currently used vehicle provides.","These context-dependent profiles are preferably defined for a user individually according to a type of vehicle.","It is possible, for example, for a “first user” profile to be present which explains innovations only a single time, while a “second user” profile explains these changes within a specific time period whenever a journey is begun.","It is very advantageous to define context dependent profiles which output options and/or data as a function of at least one current vehicle variable, such as for example, currently measured sensor data or vehicle system states such as the heating level of the seat heating system or the range of the vehicle with the currently available quantity of fuel which is determined by the trip computer.","As a result, a vehicle user can be informed particularly quickly about specific situations because, data corresponding to the situation indicated by the vehicle variables is issued to him, as a function of the profiles.","For example, options or data relating to a TCS (Traction Control System) can be output if the wheels spin during the process of driving off.","It is also possible to output, as a function of the profiles, instructions on “cutting free” (by the user briefly switching off the TCS during the process of driving off), by virtue of the fact that the operator control element for switching off the TCS is pointed out and/or direct triggering—i.e.","deactivation—with or without previous interrogation of the user is brought about.","In the case of a flat tire or an accident it is possible, for example, to issue options or data on the correct behavior in emergency situations and/or on the position of the warning triangle in the vehicle.","The vehicle variables can also be transmitted from outside the vehicle, for example in wireless fashion by XFCD (“Extended Floating Car Data”).","It is thus possible to transmit into the vehicle sensor data about weather conditions on the route ahead.","The user is then issued with options or data as a function of the profiles such that the user can set an air conditioning system in his vehicle in a way which is advantageous for these conditions.","A further advantageous embodiment of the invention provides for a plausibility check for a control operation which is carried out by the user, with the options and/or the data being output as a function of the result of the plausibility check.","This increases the operator control reliability and increases the convenience because a user is offered information about an operator control function or its operator control element as a function of the profile if he makes an operator control error or if an algorithm detects an operator control problem.","As a result, it is possible, for example when there is multiple use of the operator control elements for main beam and/or indicator lights within a short period of time while it is raining at the same time, to offer and/or issue data on the operator control of the windshield wiper, in particular on how to switch it on, as options for selection.","In a further configuration of the invention, vehicle variables and/or operator control operations are stored in a memory, which is initially interrogated before options are represented and/or data is output as a function of the profiles.","If values relating to preceding vehicle variables or operator control operations are stored in the memory, these values are checked to determine whether they are to be taken into account in the representation of the current options and/or in the outputting of the current data.","This further increases operating convenience because a rapid selection of options and/or data is matched individually to the user.","As it were, a “memory” is provided by virtue of the fact that conclusions can be drawn relating to a current operator control operation, for example from analyzing preceding operator control operations.","In the case of operator control steps which the user usually makes, it is thus possible to suppress a representation of information and outputting of data.","The classification of operator control problems or the categorization of current vehicle variables is also improved by virtue of the fact that, for example, operator control steps which are specific to a user are detected and no longer classified as an operator control problem and explanatory data is output.","Such a recognition can be achieved, for example, by using the methods of “artificial intelligence” (such as a neural network).","By checking of values stored in the memory it is also possible to make modifications to the currently defined, context-dependent profile itself and/or retrieve and subsequently define a different context dependent profile.","It is advantageous if at least part of an operating manual for the motor vehicle is represented as information relating to the motor vehicle.","A method suitable for this purpose is described, for example, in the German Patent Application DE 199 59 755.3-21 whose content is included herein by reference.","In this method, a current equipment variant of the motor vehicle is determined in a preceding step and a second database is generated from a first database as a function of this determined variant, this second database comprising only information on the operator control for the current equipment variant of the motor vehicle.","In this way, a continuously updated information state relating to the motor vehicle and its operator control elements is ensured.","The device according to the invention for processing and outputting data relating to information regarding a motor vehicle or its operator control elements comprises a display for representing options relating to the information (referred to below for short as first output means), a display for the user to select at least one option (referred to below for short as selection means), a display for outputting data relating to the selected option (referred to for short below as second output means) and a processing unit which generates, by reference to at least one defined, context-dependent profile, a number of options that are characterized by the profile and which processing unit outputs data as a function of the profiles and options.","This processing unit may be embodied, for example, as a vehicle computer (a processing unit which is arranged in the vehicle).","However, it can also be implemented by providing additional functions to a unit which is already present in the vehicle—for example a control device.","The first output means and/or the second output means may be, for example, display units of conventional design such as LCD (Liquid Crystal Display), or may be presented on the windshield (Head Up Display).","It is advantageous here to embody the display unit as a function of the set of characteristics of the options and/or data (both of these are referred to below as data set).","For example, a small data set may be output by means of a display which is already present in the vehicle, while a relatively large data set is output by means of a further relatively large display which is mounted on the dashboard or in the center console.","In addition, further possibilities for displaying options and/or data are available.","The outputting of the options and data can also depend on what technical equipment a vehicle has.","For example, when a suitable display fails, it is possible to suppress the outputting of certain data (for example video sequences forming a short introduction to the vehicle for a new user).","Of course, it is also possible to provide a joint display for outputting the options and data.","In order to integrate the first and/or second output means with the selection means, a conventional touch-sensitive display (touch screen) is preferably provided.","Alternatively or additionally, the selection means may be embodied as a voice operated controller, such as is already used today in motor vehicles, permitting the driver to concentrate his visual attention on the events on the road.","Furthermore, the selection means may be embodied as a manual operator control element of conventional design, for example, in the form of a joystick, or as a combined rotary/pushbutton knob which has in each case a different number of latching positions depending on the number of options for the information.","Such an operator control element is also already present in many vehicles (for example for controlling a navigation device), and can be used advantageously.","Further possible operator control systems may be provided.","In order to output the options or the selected data it is possible to provide means of a conventional design, for example audible outputs, such as an audio system, or haptic output (for example a force feedback operator control element, an opposing force being exerted at said operator control element when it is used).","The means which is used for inputting and/or outputting may also be defined in a variable fashion as a function of the profiles, for example a visual output is replaced by an audible output as soon as the vehicle exceeds a defined limiting speed.","Furthermore, the data which are output can be further processed and fed to other systems, in particular assistance systems in a motor vehicle.","Other objects, advantages and novel features of the present invention will become apparent from the following detailed description of the invention when considered in conjunction with the accompanying drawings."],["This application claims the priority of German patent document 101 17 410.1, filed Apr.","6, 2001 (PCT No.","PCT/EP02/01485) filed Feb. 13, 2002), the disclosure of which is expressly incorporated by reference herein.","BACKGROUND AND SUMMARY OF THE INVENTION The invention relates to a method and apparatus for outputting data providing information regarding a motor vehicle to a user of the motor vehicle.","International Patent Document WO 98/34812 discloses a multimedia information and control system for a motor vehicle, in which the user selects the information of interest to him from a defined global set.","Various options relating to the information are then offered, from which he can select at least one option and cause the respective data to be output.","Furthermore, an operator control function which corresponds to the data that are output can be executed.","One object of the invention is to provide an improved method and apparatus of the generic type in which operating convenience for the user is increased and operating safety and traffic safety are improved.","This and other objects and advantages are achieved by the method and apparatus, according to the invention, in which the type and number of the options generated with respect to the information is determined and the respective data are output, as a function of at least one context-dependent profile, thus providing the user with fast and simple access to data which are more wide ranging in that they describe the individual information more closely.","As used herein, the word “information” designates a generic classification or heading under which various data can be grouped, for example “windshield wiper”.","The data relating to particular information comprises assigned details which are stored in a memory and which can be output in different ways for the user: in the case of “windshield wiper”, for example, an introduction video for operator control or a brief audio explanation of the interval levels.","The context-dependent profile relates to the characterization of at least one secondary factor in the interaction between the vehicle, the vehicle user and the environment (for example the user's age).","By reference to the context dependent profile, the options which are associated with the “information” (e.g., windshield wipers) are conditioned in such a way that from a total set of options, only those options which are assigned to the context-dependent profile are generated.","Furthermore, by using the context-dependent profile it is also possible to output data automatically, without the user's previously having selected an option.","By means of the context-dependent profile the number of generated options from which the user can make a selection and the outputting of the data are determined in such a manner that a clearly organized and user-compatible representation is made possible.","Thus, when an item of information relating to the motor vehicle is requested, a subset of the entire set of data relating to this information is selected as a function of the profiles.","(The subset can of course also correspond to the entire data set.)","The selected data are output in a way which is particularly data-optimized for the user.","As a result, the sequence of selecting data and outputting data is shortened, depending on the context.","The invention also makes a contribution both to convenience and to safety.","For example, when a vehicle display lights up, the user is automatically offered an explanation of this vehicle display as a function of the profile, by reason of appropriate data being output.","The user is thus immediately informed, without having to think about the explanation or even perform awkward control operations.","Likewise it possible to provide the user with selective proposals as a function of the profiles and to instruct him about vehicle functions which are suitable to use in a specific situation.","It is advantageous if an operator control function whose use is proposed with the data which is output, is triggered as a function of the profiles and options.","As a result, the operating convenience is increased further, because the user does not need to search for the triggering means which are assigned to this operator control function by virtue of the fact that the proposed operator control function is triggered in an easy way which is optimized as much as possible with respect to time.","Such direct triggering of the operator control function, without previous interrogation of the user makes it possible to trigger the operator control function much more reliably in particular in driving situations for which timing is critical.","Alternatively, direct triggering of the operator control function may be provided.","In this case, as a function of the profiles, a corresponding option is defined, said option being selected and then bringing about indirect triggering of the operator control function.","An example of direct or indirect triggering occurs if the driver selects the reverse gear on his vehicle, and as a result, either an electrically movable rear roller blind is retracted (direct triggering) or the retraction of the rear roller blind is proposed as an option (indirect triggering).","One advantageous embodiment of the invention selects, for a particular user, a context-dependent profile based on the assignment of that user to a defined user group.","As a result, convenience is increased further in that a specific, group related information requirement is fulfilled.","For example, the assignment of the user to a user group takes into account whether he or she is already familiar with the vehicle or the make of vehicle.","For example, a first user group may comprise users for which the vehicle or the make of vehicle is new, and a second user group may comprise experienced users.","If, for example, a user is assigned to the first user group, a specific context dependent profile is defined, bringing about an additional, explanatory outputting of data, for example relating to operator control elements.","Alternatively or additionally, a brief explanation on the vehicle and its operator control elements is possible when the user is assigned to the first user group.","The described assignment to user groups is carried out, for example, during visits for servicing of the vehicle.","Thus, when the vehicle is handed back the “new user” profile can then be defined.","On one of the subsequent visits to the workshop it is then possible to define the “experienced users” group.","A further possibility of a predefinable user group is to select an “interested user” profile which, for example with respect to predefinable vehicle reactions, supplies data in the form of explanations or technical explanations on vehicle functions.","By assigning the user to a user group for the disabled it is possible to output data as a function of the profile in order to support the user in specific situations.","For example, given an assignment of the user to a “restricted hearing” user group, as a function of the profiles, supplementary visual data are output if according to standard practice an operator control element provides audible feedback in the vehicle.","A user can also be assigned to more than one user group, so that more than one profile is assigned for him, preferably with different priorities assigned to different profiles.","If a signal for representing options and/or for outputting data (below for the sake of simplification we only refer to the outputting of options or data) from two or more profiles is present, the options or data are output in correct order and as a function of the priority of the profile.","Here, for example, the options or data with the highest priority are output immediately and additionally an option which the user can select in order to cause the further options or data to be output is illustrated.","It is also possible to define separate profiles for a plurality of users, for example for a user who drives the vehicle, a user who is a front seat passenger, or a user who is sitting in the rear of the vehicle.","In a profile which is defined for the driver of the vehicle, functions relating, for example, to driving safety are provided, while in a profile defined for the rear-seat passengers, audio and multimedia functions are provided, for example.","In this manner, the operating reliability and thus the traffic safety are increased by the context-dependent profile for the driver of the vehicle.","Similarly, for a rear-seat passenger the operating convenience is increased, and comprehensive information about the operator control functions which are available is provided.","In addition it is also thus possible to define further profiles, for example it is possible to provide a specific profile for a service technician with a representation of options and/or outputting of data on the diagnostics of the vehicle.","A further advantageous embodiment of the invention selects a context-dependent profile for a predefinable vehicle environment.","Here, a user is informed about changes which have occurred in the course of his journey by presenting options and/or data which permit the user to prepare for changed conditions on his route, as a function of the profiles.","For example, it is possible for him to obtain information about operator control functions which are suitable for the new conditions and to trigger these functions.","Thus, in the case of a road in poor condition which is stored as an attribute in the digital road map of the vehicle, it is possible to present options as a function of the profiles and/or to output data relating to the operator control elements of a ride level controller (which sets the ground clearance of the motor vehicle).","Furthermore, it is possible, for example, to define a country-specific profile.","When it is detected that a national border has been crossed, information about country-specific traffic regulations are provided as a function of the profiles.","A conventional device for locating the position of the vehicle is used, for example, to identify the border crossing.","In a further refinement of the invention, context dependent profiles are defined with respect to the type of vehicle.","As a result, a user is informed about changes which occur when the type of vehicle which he uses at a particular time changes.","If the user changes, for example, from his previous vehicle model of his make of vehicle to a newer model, only options or data which relate to operator control elements which have changed or are new in comparison to his previously used vehicle are issued to him.","This applies in an analogous way, for example, to retrofits or software updates of vehicles.","In each case the differences between the current vehicle and the vehicle which is known to the user is determined first, and only those options or data which are associated with this difference for the user are output to the user as a function of the profiles.","As a result, by virtue of the profiles, the user is informed about the possibilities which the currently used vehicle provides.","These context-dependent profiles are preferably defined for a user individually according to a type of vehicle.","It is possible, for example, for a “first user” profile to be present which explains innovations only a single time, while a “second user” profile explains these changes within a specific time period whenever a journey is begun.","It is very advantageous to define context dependent profiles which output options and/or data as a function of at least one current vehicle variable, such as for example, currently measured sensor data or vehicle system states such as the heating level of the seat heating system or the range of the vehicle with the currently available quantity of fuel which is determined by the trip computer.","As a result, a vehicle user can be informed particularly quickly about specific situations because, data corresponding to the situation indicated by the vehicle variables is issued to him, as a function of the profiles.","For example, options or data relating to a TCS (Traction Control System) can be output if the wheels spin during the process of driving off.","It is also possible to output, as a function of the profiles, instructions on “cutting free” (by the user briefly switching off the TCS during the process of driving off), by virtue of the fact that the operator control element for switching off the TCS is pointed out and/or direct triggering—i.e.","deactivation—with or without previous interrogation of the user is brought about.","In the case of a flat tire or an accident it is possible, for example, to issue options or data on the correct behavior in emergency situations and/or on the position of the warning triangle in the vehicle.","The vehicle variables can also be transmitted from outside the vehicle, for example in wireless fashion by XFCD (“Extended Floating Car Data”).","It is thus possible to transmit into the vehicle sensor data about weather conditions on the route ahead.","The user is then issued with options or data as a function of the profiles such that the user can set an air conditioning system in his vehicle in a way which is advantageous for these conditions.","A further advantageous embodiment of the invention provides for a plausibility check for a control operation which is carried out by the user, with the options and/or the data being output as a function of the result of the plausibility check.","This increases the operator control reliability and increases the convenience because a user is offered information about an operator control function or its operator control element as a function of the profile if he makes an operator control error or if an algorithm detects an operator control problem.","As a result, it is possible, for example when there is multiple use of the operator control elements for main beam and/or indicator lights within a short period of time while it is raining at the same time, to offer and/or issue data on the operator control of the windshield wiper, in particular on how to switch it on, as options for selection.","In a further configuration of the invention, vehicle variables and/or operator control operations are stored in a memory, which is initially interrogated before options are represented and/or data is output as a function of the profiles.","If values relating to preceding vehicle variables or operator control operations are stored in the memory, these values are checked to determine whether they are to be taken into account in the representation of the current options and/or in the outputting of the current data.","This further increases operating convenience because a rapid selection of options and/or data is matched individually to the user.","As it were, a “memory” is provided by virtue of the fact that conclusions can be drawn relating to a current operator control operation, for example from analyzing preceding operator control operations.","In the case of operator control steps which the user usually makes, it is thus possible to suppress a representation of information and outputting of data.","The classification of operator control problems or the categorization of current vehicle variables is also improved by virtue of the fact that, for example, operator control steps which are specific to a user are detected and no longer classified as an operator control problem and explanatory data is output.","Such a recognition can be achieved, for example, by using the methods of “artificial intelligence” (such as a neural network).","By checking of values stored in the memory it is also possible to make modifications to the currently defined, context-dependent profile itself and/or retrieve and subsequently define a different context dependent profile.","It is advantageous if at least part of an operating manual for the motor vehicle is represented as information relating to the motor vehicle.","A method suitable for this purpose is described, for example, in the German Patent Application DE 199 59 755.3-21 whose content is included herein by reference.","In this method, a current equipment variant of the motor vehicle is determined in a preceding step and a second database is generated from a first database as a function of this determined variant, this second database comprising only information on the operator control for the current equipment variant of the motor vehicle.","In this way, a continuously updated information state relating to the motor vehicle and its operator control elements is ensured.","The device according to the invention for processing and outputting data relating to information regarding a motor vehicle or its operator control elements comprises a display for representing options relating to the information (referred to below for short as first output means), a display for the user to select at least one option (referred to below for short as selection means), a display for outputting data relating to the selected option (referred to for short below as second output means) and a processing unit which generates, by reference to at least one defined, context-dependent profile, a number of options that are characterized by the profile and which processing unit outputs data as a function of the profiles and options.","This processing unit may be embodied, for example, as a vehicle computer (a processing unit which is arranged in the vehicle).","However, it can also be implemented by providing additional functions to a unit which is already present in the vehicle—for example a control device.","The first output means and/or the second output means may be, for example, display units of conventional design such as LCD (Liquid Crystal Display), or may be presented on the windshield (Head Up Display).","It is advantageous here to embody the display unit as a function of the set of characteristics of the options and/or data (both of these are referred to below as data set).","For example, a small data set may be output by means of a display which is already present in the vehicle, while a relatively large data set is output by means of a further relatively large display which is mounted on the dashboard or in the center console.","In addition, further possibilities for displaying options and/or data are available.","The outputting of the options and data can also depend on what technical equipment a vehicle has.","For example, when a suitable display fails, it is possible to suppress the outputting of certain data (for example video sequences forming a short introduction to the vehicle for a new user).","Of course, it is also possible to provide a joint display for outputting the options and data.","In order to integrate the first and/or second output means with the selection means, a conventional touch-sensitive display (touch screen) is preferably provided.","Alternatively or additionally, the selection means may be embodied as a voice operated controller, such as is already used today in motor vehicles, permitting the driver to concentrate his visual attention on the events on the road.","Furthermore, the selection means may be embodied as a manual operator control element of conventional design, for example, in the form of a joystick, or as a combined rotary/pushbutton knob which has in each case a different number of latching positions depending on the number of options for the information.","Such an operator control element is also already present in many vehicles (for example for controlling a navigation device), and can be used advantageously.","Further possible operator control systems may be provided.","In order to output the options or the selected data it is possible to provide means of a conventional design, for example audible outputs, such as an audio system, or haptic output (for example a force feedback operator control element, an opposing force being exerted at said operator control element when it is used).","The means which is used for inputting and/or outputting may also be defined in a variable fashion as a function of the profiles, for example a visual output is replaced by an audible output as soon as the vehicle exceeds a defined limiting speed.","Furthermore, the data which are output can be further processed and fed to other systems, in particular assistance systems in a motor vehicle.","Other objects, advantages and novel features of the present invention will become apparent from the following detailed description of the invention when considered in conjunction with the accompanying drawings.","BRIEF DESCRIPTION OF THE DRAWINGS FIG.","1 shows a schematic block circuit diagram of an embodiment of the device for outputting data relating to information regarding a motor vehicle; FIG.","2 shows a schematic block circuit diagram of a further embodiment of the device; FIG.","3 shows a plan view of an embodiment of the device; FIG.","4 shows, by way of example, an image which is represented on a display unit, and is generated by the device in a basic state; FIG.","5 shows, by way of example, an image which is represented on a display unit, and is generated by the device in a state with a preselected option; and FIG.","6 shows, by way of example, an image which is represented on a display unit, and is generated by the device in a state with output data.","DETAILED DESCRIPTION OF THE DRAWINGS FIG.","1 shows a motor vehicle 1 with a device 2 for outputting data (D) with a processing unit 3.The device 2 comprises a means 4 for representing options (On) relating to the information (I) (referred to below for short as options output means 4), and a means 5 for outputting data (D) relating to the selected option (O) (referred to below for short as data output means 5).","The two output means 4 and 5 are, for example, two displays which are arranged at a distance from one another.","Alternatively, both output means 4 and 5 can be formed by a display.","As a further alternative or additionally, an audible output is provided for the output means 4 and/or 5.Furthermore, the processing unit 3 has a means 6 (referred to below for short as selection means 6) by which the user can select at least one option (O).","The selection means 6 is embodied as a separate unit, for example, as a voice operated controller.","Additionally or alternatively, a manual operator control element of conventional design may be provided, such as for example a joystick or a combined rotary/pushbutton knob, which has a large number of different latched positions depending on the number of options (On) for the information.","As a further alternative it is possible to provide a unit which detects body movements or facial expression of the user, classified them and assigns them to a corresponding operator control request, for which the requested operator control operation is then carried out.","The processing unit 3 is supplied with values (W) from at least one component 7, such as a vehicle-mounted sensor which measures the current speed, or a control unit which is provided for setting possibilities of the vehicle seat.","However, it is also possible to transmit values in a wireless fashion into the vehicle, for example from other vehicles and/or from local transmission devices arranged at the edge of the road pavement.","Furthermore, a unit 8 for storing values (Wn) such as, for example, sensor data and/or operator control operations is provided (referred to below as storage unit 8).","The storage unit 8 may be embodied as part of the usual peripheral components of the processing unit 3 with a conventional design.","Furthermore, at least one context-dependent profile (P) is selected from a memory 9 by at least one profile selection means 10, and fed to the processing unit 3.The profile selection means 10 may provide a switch, for example a rotary or pushbutton switch, which is suitable for this.","The user places the switch in a switch position which is provided for it and which is sensed by the processing unit 3, after which it retrieves the context-dependent profile or profiles (P) which is selected by the user from the memory 9 and allocates them to the user.","Alternatively, the profile selection means 10 is connected to the starting device of the engine as a fingerprint sensor (detection of the “fingerprint” of the user).","When the user initiates an engine starting procedure, he is identified by the profile selection means 10.Then, at least one context dependent profile (P) which is provided for the user is retrieved from the memory 9 and defined for him.","Likewise, it is possible to provide a specific interface of the processor unit 3 as the profile selection means 10, to which interface, for example, a customary diagnostic device for defining a context dependent profile (P) can be connected.","Alternatively or additionally, an electronic data carrier can be provided for the combination of memory 9 and profile selection means 10, the user carrying said data carrier, for example a driver-specific chip card, on him and introducing it into a device which is connected to the vehicle.","The user-specific values which are evaluated by the processing unit 3 and bring about the selection of a context-dependent profile (P) which is suitable for the user are stored on the chip card.","The electronic data carrier can also be used for storing other values.","For example, the data carrier can also be provided for storing values (Wn) such as, for example, sensor data and/or operator control operations of the user.","Alternatively or additionally, the memory 9 can also be provided outside the vehicle for example, in a service center (a central processing unit with a database which can be used in a wireless fashion by subscribers to a respective service).","A context-dependent profile (P) which is provided for the user is transferred from the service center into the vehicle in a wireless fashion if it is detected in this control center that a transceiver unit (“radio key”) which is carried by the user initiates vehicle starting procedure.","Alternatively or additionally, the service center is also used to store vehicle-related values (Wn) such as, for example, sensor data and/or operator control operations of the user.","The selection of the information (I), the type and number of options (On) generated with respect to the information (I) and the outputting of the respective data (D) are controlled by the context-dependent profile (P) by means of the processing unit 3.The data (D) which are to be output are selected from a database 11 as a function of profiles and/or options, read and output using the output means 5.Depending on the type and design of the device 2, the database 11 is integrated in the motor vehicle 1.Alternatively or additionally, data from an external database 12 can be read by the processing device 2 via wireless transmission R. This external database 12 can be provided in the service center described above.","FIG.","2 illustrates a further embodiment of the device 2 for outputting data (D) relating to the information (I).","The device 2 comprises two separate networks 13, 14, which are embodied as units of network nodes that are each connected to a plurality of transmission lines.","This distributed design divides the functional units of the device 2 (shown in FIG.","1) into the network 13 in which the values (W) of the component 7 are collected and conditioned, while storage, conditioning and outputting of data (D) is carried out by the network 14.The networks 13, 14 are connected to one another by a transmission line 15.In order to connect the two networks 13, 14 to the transmission line 15, the gateway network node 16 is provided in the network 13 and the gateway network node 17 is provided in the network 14.As is schematically illustrated in FIG.","2, the two networks 13, 14 have different architectures and are thus configured for different requirements.","The network 13 has a tree structure and comprises components 7, the storage unit 8, the memory 9 and an associated display unit 18, while the network 14, which has a ring structure and is suitable particularly for transmitting large amounts of data, comprises the database 11 and an associated, large-format display unit 19.FIG.","3 is a plan view of an embodiment of the device 2 in the dashboard of the motor vehicle 1, including a display unit 18, the selection means 6 for selecting options (On) and the display unit 19.The processing unit 3 causes data (D) relating to information (I) to be represented on the display unit 18, as a function of profiles and/or options.","At the same time, the information (I) designates a global term which can include various types of data (D).","As is illustrated in block 20, explanations of warning displays and changes of states of the motor vehicle 1 as well as of exceptional situations and suitable operator control functions are output on the display unit 18 by the processing unit 3.When there is a more wide ranging information request relating to data (D) which has already been output in the display unit 18 in the form of brief information, further data (D) is output as a detailed explanation in the display unit 19 as a function of the profiles.","The display units 18, 19 are appropriately embodied in terms of their size, type and arrangement in this respect.","Additionally or alternatively, options (O) relating to the information (I) for the outputting of data are represented on the display unit 18, as illustrated by way of example in FIG.","4.A respective option (Os) for the outputting of the data (D) which describe the respective information is selected by operating the selection means 6.When there is a direct request for information (I) by the user via the manual operated control element 6, corresponding data (D) are output on the display unit 19.This data comprise, as shown in block 22, an online help, a keyword/subject search feature, an initial instruction feature and an explanation of operator control switches, as shown in FIG.","3.FIG.","4 shows by way of example an image which is represented on the output means 4 or 5 and generated by the device 2 as a function of the profiles when the device 2 is in a basic state.","In this state, the user can request data (D) directly as a function of the profiles.","As shown in FIG.","4 as a function of the context dependent profile, the user is provided, with the options 01 to 07 relating to the information (I) “motor vehicle information system”.","The user can retrieve explanatory data (D) and/or further options (O) relating to the multipurpose instrument and to the central unit by selecting corresponding options 01, 02 by using the selection means 6.Here, a further option 08, 09 relating to the two first options 01, 02, which can be selected by selection, is already defined.","The option 01 is already selected as a function of the profiles.","Furthermore, the user can selectively search for subjects on which he desires further explanations by selecting the option 03 “subject search”.","If the user selects the option 04 “explanation of switch”, he can cause data (D) and/or further options (O) relating to individual operator control elements to be output directly.","The selection of the option 05 “initial instruction” by the user causes data (D) and/or further options (O) to be output which are advantageous for the user if he is using the motor vehicle for the first time.","By selecting the option 06 “last subject” data which the user had already requested once at an earlier time is output to the user.","Selecting the option 07 “settings” provides the user with information on the device according to the invention itself.","FIG.","5 shows by way of example a further image which is represented on the output means 4 or 5.The image represents to the user that the device 2 is in a state in which an option (O) has already been selected.","In this state, the user can cause the corresponding data (D) to be output automatically as a function of the profiles, and as a function of options by selecting the respective option (O).","Likewise, a number of symbols A indicate to the user how many options 0 have already been selected.","Here, the respectively generated images are depicted one on top of the other as a hierarchical structure.","The direct representation of such an image as a function of the profiles according to the invention avoids the tedious selection process which would otherwise precede a representation of this image.","Directly selecting a symbol A causes the respective image to be represented directly in the hierarchical structure.","A symbol (W) indicates to the user that further options (O) have been generated, and these can be “scrolled through” by selecting the symbol (W).","FIG.","6 shows by way of example an image which is represented on the display unit 5.This image is generated by the device 2 if it is in a state with data (D) which have been output.","As is apparent in FIG.","6, these data (D) comprise a pictogram (P), which has been obtained from the operating manual of the motor vehicle 1, and explanatory text.","Selecting the option 011 causes further data (D) to be output.","Selecting the option 012 causes thematically related data (D) to be represented as options (O) for outputting.","Selecting the option 013 causes the operator control function which is explained by the data (D) to be triggered.","Selecting the option 014 causes the image generated by a preceding processing step to be represented.","The foregoing disclosure has been set forth merely to illustrate the invention and is not intended to be limiting.","Since modifications of the disclosed embodiments incorporating the spirit and substance of the invention may occur to persons skilled in the art, the invention should be construed to include everything within the scope of the appended claims and equivalents thereof."]],"string":"[\n [\n \"Method and divece for outputting data relating to information of a motor vehicle\",\n \"In a method for outputting data relating to user selectable information regarding a motor vehicle, a plurality of user selectable options relating to the information are represented, and data relating to the selected option is output.\",\n \"At least one context-dependent profile is defined, and a set of options determined by the profile is generated.\",\n \"Data are output as a function of the profiles and options.\"\n ],\n [\n \"1-15.\",\n \"(canceled) 16.A method for outputting to a vehicle user, data relating to information concerning a motor vehicle, said method comprising: defining at least one context-dependent profile; generating user selectable options for at least one item of information, which options are characterized by the at least one context-dependent profile; and outputting data relating to a selected option, said data being selected from an entire data set as a function of the at least one profile.\",\n \"17.The method as claimed in claim 16, wherein an operator control function which corresponds to the data which are output is executed as a function of the profiles and options.\",\n \"18.The method as claimed in claim 16, wherein a particular profile is provided for a predefinable user group.\",\n \"19.The method as claimed in claim 16, wherein a particular profile is provided for a predefinable vehicle environment.\",\n \"20.The method as claimed in claim 16, wherein a particular profile is provided based on the type of vehicle.\",\n \"21.The method as claimed in claim 16, wherein the options are presented and/or the data are output as a function of current vehicle parameters.\",\n \"22.The method as claimed in claim 16, further comprising: performing a plausibility check for a control operation that is carried out by the vehicle user; and presenting the options or outputting the data are output as a function of a result of said check.\",\n \"23.The method as claimed in claim 16, further comprising: storing preceding vehicle variables and/or preceding control operations; and presenting options or outputting the data, as a function of stored values.\",\n \"24.The method as claimed in claim 16, further comprising: processing the data further; and feeding the data to other systems in the motor vehicle.\",\n \"25.The method as claimed in claim 16, wherein at least part of a motor vehicle operating manual is presented as information.\",\n \"26.A device for outputting to a vehicle user, data relating to information concerning a motor vehicle, said device comprising: a processing unit which generates, by reference to at least one defined context-dependent profile, a plurality of options which are associated with the at least one profile; interface means for representing the options generated by the processing unit; means for a user selecting at least one option from among the options generated by the processing unit and represented by the interface means; and means for outputting data relating to a selected option, as a function of the at least one profile.\",\n \"27.The device as claimed in claim 26, further comprising at least one display unit for outputting data as a function of a data set which characterizes it.\",\n \"28.The device as claimed in claim 26, wherein the selection means comprises a voice input.\",\n \"29.The device as claimed in claim 26, wherein the selection means comprises a manual operator control element.\",\n \"30.The device as claimed in claim 26, wherein means are provided for audibly outputting options and/or data.\",\n \"31.A method for selectively outputting vehicle related information to a vehicle user, comprising: providing an overall data set that includes data relating to said vehicle, said data being accessible according to a hierarchical classification structure, including a plurality of items of information, each of said items of information having associated therewith selected data from said overall data set, said selected data being selectively displayable according to a plurality of selectable options associated with said information; defining a plurality of context dependent profiles whose content is based on parameters characterizing at least one of said vehicle, its operation and said vehicle user; in response to designation of an item of information, generating a set of selectable options from among said plurality of selectable options, based on a particular context dependent profile; and outputting data relating to a selected option, as a function of the particular option.\"\n ],\n [\n \"<SOH> BACKGROUND AND SUMMARY OF THE INVENTION <EOH>The invention relates to a method and apparatus for outputting data providing information regarding a motor vehicle to a user of the motor vehicle.\",\n \"International Patent Document WO 98/34812 discloses a multimedia information and control system for a motor vehicle, in which the user selects the information of interest to him from a defined global set.\",\n \"Various options relating to the information are then offered, from which he can select at least one option and cause the respective data to be output.\",\n \"Furthermore, an operator control function which corresponds to the data that are output can be executed.\",\n \"One object of the invention is to provide an improved method and apparatus of the generic type in which operating convenience for the user is increased and operating safety and traffic safety are improved.\",\n \"This and other objects and advantages are achieved by the method and apparatus, according to the invention, in which the type and number of the options generated with respect to the information is determined and the respective data are output, as a function of at least one context-dependent profile, thus providing the user with fast and simple access to data which are more wide ranging in that they describe the individual information more closely.\",\n \"As used herein, the word “information” designates a generic classification or heading under which various data can be grouped, for example “windshield wiper”.\",\n \"The data relating to particular information comprises assigned details which are stored in a memory and which can be output in different ways for the user: in the case of “windshield wiper”, for example, an introduction video for operator control or a brief audio explanation of the interval levels.\",\n \"The context-dependent profile relates to the characterization of at least one secondary factor in the interaction between the vehicle, the vehicle user and the environment (for example the user's age).\",\n \"By reference to the context dependent profile, the options which are associated with the “information” (e.g., windshield wipers) are conditioned in such a way that from a total set of options, only those options which are assigned to the context-dependent profile are generated.\",\n \"Furthermore, by using the context-dependent profile it is also possible to output data automatically, without the user's previously having selected an option.\",\n \"By means of the context-dependent profile the number of generated options from which the user can make a selection and the outputting of the data are determined in such a manner that a clearly organized and user-compatible representation is made possible.\",\n \"Thus, when an item of information relating to the motor vehicle is requested, a subset of the entire set of data relating to this information is selected as a function of the profiles.\",\n \"(The subset can of course also correspond to the entire data set.)\",\n \"The selected data are output in a way which is particularly data-optimized for the user.\",\n \"As a result, the sequence of selecting data and outputting data is shortened, depending on the context.\",\n \"The invention also makes a contribution both to convenience and to safety.\",\n \"For example, when a vehicle display lights up, the user is automatically offered an explanation of this vehicle display as a function of the profile, by reason of appropriate data being output.\",\n \"The user is thus immediately informed, without having to think about the explanation or even perform awkward control operations.\",\n \"Likewise it possible to provide the user with selective proposals as a function of the profiles and to instruct him about vehicle functions which are suitable to use in a specific situation.\",\n \"It is advantageous if an operator control function whose use is proposed with the data which is output, is triggered as a function of the profiles and options.\",\n \"As a result, the operating convenience is increased further, because the user does not need to search for the triggering means which are assigned to this operator control function by virtue of the fact that the proposed operator control function is triggered in an easy way which is optimized as much as possible with respect to time.\",\n \"Such direct triggering of the operator control function, without previous interrogation of the user makes it possible to trigger the operator control function much more reliably in particular in driving situations for which timing is critical.\",\n \"Alternatively, direct triggering of the operator control function may be provided.\",\n \"In this case, as a function of the profiles, a corresponding option is defined, said option being selected and then bringing about indirect triggering of the operator control function.\",\n \"An example of direct or indirect triggering occurs if the driver selects the reverse gear on his vehicle, and as a result, either an electrically movable rear roller blind is retracted (direct triggering) or the retraction of the rear roller blind is proposed as an option (indirect triggering).\",\n \"One advantageous embodiment of the invention selects, for a particular user, a context-dependent profile based on the assignment of that user to a defined user group.\",\n \"As a result, convenience is increased further in that a specific, group related information requirement is fulfilled.\",\n \"For example, the assignment of the user to a user group takes into account whether he or she is already familiar with the vehicle or the make of vehicle.\",\n \"For example, a first user group may comprise users for which the vehicle or the make of vehicle is new, and a second user group may comprise experienced users.\",\n \"If, for example, a user is assigned to the first user group, a specific context dependent profile is defined, bringing about an additional, explanatory outputting of data, for example relating to operator control elements.\",\n \"Alternatively or additionally, a brief explanation on the vehicle and its operator control elements is possible when the user is assigned to the first user group.\",\n \"The described assignment to user groups is carried out, for example, during visits for servicing of the vehicle.\",\n \"Thus, when the vehicle is handed back the “new user” profile can then be defined.\",\n \"On one of the subsequent visits to the workshop it is then possible to define the “experienced users” group.\",\n \"A further possibility of a predefinable user group is to select an “interested user” profile which, for example with respect to predefinable vehicle reactions, supplies data in the form of explanations or technical explanations on vehicle functions.\",\n \"By assigning the user to a user group for the disabled it is possible to output data as a function of the profile in order to support the user in specific situations.\",\n \"For example, given an assignment of the user to a “restricted hearing” user group, as a function of the profiles, supplementary visual data are output if according to standard practice an operator control element provides audible feedback in the vehicle.\",\n \"A user can also be assigned to more than one user group, so that more than one profile is assigned for him, preferably with different priorities assigned to different profiles.\",\n \"If a signal for representing options and/or for outputting data (below for the sake of simplification we only refer to the outputting of options or data) from two or more profiles is present, the options or data are output in correct order and as a function of the priority of the profile.\",\n \"Here, for example, the options or data with the highest priority are output immediately and additionally an option which the user can select in order to cause the further options or data to be output is illustrated.\",\n \"It is also possible to define separate profiles for a plurality of users, for example for a user who drives the vehicle, a user who is a front seat passenger, or a user who is sitting in the rear of the vehicle.\",\n \"In a profile which is defined for the driver of the vehicle, functions relating, for example, to driving safety are provided, while in a profile defined for the rear-seat passengers, audio and multimedia functions are provided, for example.\",\n \"In this manner, the operating reliability and thus the traffic safety are increased by the context-dependent profile for the driver of the vehicle.\",\n \"Similarly, for a rear-seat passenger the operating convenience is increased, and comprehensive information about the operator control functions which are available is provided.\",\n \"In addition it is also thus possible to define further profiles, for example it is possible to provide a specific profile for a service technician with a representation of options and/or outputting of data on the diagnostics of the vehicle.\",\n \"A further advantageous embodiment of the invention selects a context-dependent profile for a predefinable vehicle environment.\",\n \"Here, a user is informed about changes which have occurred in the course of his journey by presenting options and/or data which permit the user to prepare for changed conditions on his route, as a function of the profiles.\",\n \"For example, it is possible for him to obtain information about operator control functions which are suitable for the new conditions and to trigger these functions.\",\n \"Thus, in the case of a road in poor condition which is stored as an attribute in the digital road map of the vehicle, it is possible to present options as a function of the profiles and/or to output data relating to the operator control elements of a ride level controller (which sets the ground clearance of the motor vehicle).\",\n \"Furthermore, it is possible, for example, to define a country-specific profile.\",\n \"When it is detected that a national border has been crossed, information about country-specific traffic regulations are provided as a function of the profiles.\",\n \"A conventional device for locating the position of the vehicle is used, for example, to identify the border crossing.\",\n \"In a further refinement of the invention, context dependent profiles are defined with respect to the type of vehicle.\",\n \"As a result, a user is informed about changes which occur when the type of vehicle which he uses at a particular time changes.\",\n \"If the user changes, for example, from his previous vehicle model of his make of vehicle to a newer model, only options or data which relate to operator control elements which have changed or are new in comparison to his previously used vehicle are issued to him.\",\n \"This applies in an analogous way, for example, to retrofits or software updates of vehicles.\",\n \"In each case the differences between the current vehicle and the vehicle which is known to the user is determined first, and only those options or data which are associated with this difference for the user are output to the user as a function of the profiles.\",\n \"As a result, by virtue of the profiles, the user is informed about the possibilities which the currently used vehicle provides.\",\n \"These context-dependent profiles are preferably defined for a user individually according to a type of vehicle.\",\n \"It is possible, for example, for a “first user” profile to be present which explains innovations only a single time, while a “second user” profile explains these changes within a specific time period whenever a journey is begun.\",\n \"It is very advantageous to define context dependent profiles which output options and/or data as a function of at least one current vehicle variable, such as for example, currently measured sensor data or vehicle system states such as the heating level of the seat heating system or the range of the vehicle with the currently available quantity of fuel which is determined by the trip computer.\",\n \"As a result, a vehicle user can be informed particularly quickly about specific situations because, data corresponding to the situation indicated by the vehicle variables is issued to him, as a function of the profiles.\",\n \"For example, options or data relating to a TCS (Traction Control System) can be output if the wheels spin during the process of driving off.\",\n \"It is also possible to output, as a function of the profiles, instructions on “cutting free” (by the user briefly switching off the TCS during the process of driving off), by virtue of the fact that the operator control element for switching off the TCS is pointed out and/or direct triggering—i.e.\",\n \"deactivation—with or without previous interrogation of the user is brought about.\",\n \"In the case of a flat tire or an accident it is possible, for example, to issue options or data on the correct behavior in emergency situations and/or on the position of the warning triangle in the vehicle.\",\n \"The vehicle variables can also be transmitted from outside the vehicle, for example in wireless fashion by XFCD (“Extended Floating Car Data”).\",\n \"It is thus possible to transmit into the vehicle sensor data about weather conditions on the route ahead.\",\n \"The user is then issued with options or data as a function of the profiles such that the user can set an air conditioning system in his vehicle in a way which is advantageous for these conditions.\",\n \"A further advantageous embodiment of the invention provides for a plausibility check for a control operation which is carried out by the user, with the options and/or the data being output as a function of the result of the plausibility check.\",\n \"This increases the operator control reliability and increases the convenience because a user is offered information about an operator control function or its operator control element as a function of the profile if he makes an operator control error or if an algorithm detects an operator control problem.\",\n \"As a result, it is possible, for example when there is multiple use of the operator control elements for main beam and/or indicator lights within a short period of time while it is raining at the same time, to offer and/or issue data on the operator control of the windshield wiper, in particular on how to switch it on, as options for selection.\",\n \"In a further configuration of the invention, vehicle variables and/or operator control operations are stored in a memory, which is initially interrogated before options are represented and/or data is output as a function of the profiles.\",\n \"If values relating to preceding vehicle variables or operator control operations are stored in the memory, these values are checked to determine whether they are to be taken into account in the representation of the current options and/or in the outputting of the current data.\",\n \"This further increases operating convenience because a rapid selection of options and/or data is matched individually to the user.\",\n \"As it were, a “memory” is provided by virtue of the fact that conclusions can be drawn relating to a current operator control operation, for example from analyzing preceding operator control operations.\",\n \"In the case of operator control steps which the user usually makes, it is thus possible to suppress a representation of information and outputting of data.\",\n \"The classification of operator control problems or the categorization of current vehicle variables is also improved by virtue of the fact that, for example, operator control steps which are specific to a user are detected and no longer classified as an operator control problem and explanatory data is output.\",\n \"Such a recognition can be achieved, for example, by using the methods of “artificial intelligence” (such as a neural network).\",\n \"By checking of values stored in the memory it is also possible to make modifications to the currently defined, context-dependent profile itself and/or retrieve and subsequently define a different context dependent profile.\",\n \"It is advantageous if at least part of an operating manual for the motor vehicle is represented as information relating to the motor vehicle.\",\n \"A method suitable for this purpose is described, for example, in the German Patent Application DE 199 59 755.3-21 whose content is included herein by reference.\",\n \"In this method, a current equipment variant of the motor vehicle is determined in a preceding step and a second database is generated from a first database as a function of this determined variant, this second database comprising only information on the operator control for the current equipment variant of the motor vehicle.\",\n \"In this way, a continuously updated information state relating to the motor vehicle and its operator control elements is ensured.\",\n \"The device according to the invention for processing and outputting data relating to information regarding a motor vehicle or its operator control elements comprises a display for representing options relating to the information (referred to below for short as first output means), a display for the user to select at least one option (referred to below for short as selection means), a display for outputting data relating to the selected option (referred to for short below as second output means) and a processing unit which generates, by reference to at least one defined, context-dependent profile, a number of options that are characterized by the profile and which processing unit outputs data as a function of the profiles and options.\",\n \"This processing unit may be embodied, for example, as a vehicle computer (a processing unit which is arranged in the vehicle).\",\n \"However, it can also be implemented by providing additional functions to a unit which is already present in the vehicle—for example a control device.\",\n \"The first output means and/or the second output means may be, for example, display units of conventional design such as LCD (Liquid Crystal Display), or may be presented on the windshield (Head Up Display).\",\n \"It is advantageous here to embody the display unit as a function of the set of characteristics of the options and/or data (both of these are referred to below as data set).\",\n \"For example, a small data set may be output by means of a display which is already present in the vehicle, while a relatively large data set is output by means of a further relatively large display which is mounted on the dashboard or in the center console.\",\n \"In addition, further possibilities for displaying options and/or data are available.\",\n \"The outputting of the options and data can also depend on what technical equipment a vehicle has.\",\n \"For example, when a suitable display fails, it is possible to suppress the outputting of certain data (for example video sequences forming a short introduction to the vehicle for a new user).\",\n \"Of course, it is also possible to provide a joint display for outputting the options and data.\",\n \"In order to integrate the first and/or second output means with the selection means, a conventional touch-sensitive display (touch screen) is preferably provided.\",\n \"Alternatively or additionally, the selection means may be embodied as a voice operated controller, such as is already used today in motor vehicles, permitting the driver to concentrate his visual attention on the events on the road.\",\n \"Furthermore, the selection means may be embodied as a manual operator control element of conventional design, for example, in the form of a joystick, or as a combined rotary/pushbutton knob which has in each case a different number of latching positions depending on the number of options for the information.\",\n \"Such an operator control element is also already present in many vehicles (for example for controlling a navigation device), and can be used advantageously.\",\n \"Further possible operator control systems may be provided.\",\n \"In order to output the options or the selected data it is possible to provide means of a conventional design, for example audible outputs, such as an audio system, or haptic output (for example a force feedback operator control element, an opposing force being exerted at said operator control element when it is used).\",\n \"The means which is used for inputting and/or outputting may also be defined in a variable fashion as a function of the profiles, for example a visual output is replaced by an audible output as soon as the vehicle exceeds a defined limiting speed.\",\n \"Furthermore, the data which are output can be further processed and fed to other systems, in particular assistance systems in a motor vehicle.\",\n \"Other objects, advantages and novel features of the present invention will become apparent from the following detailed description of the invention when considered in conjunction with the accompanying drawings.\"\n ],\n [\n \"<SOH> BACKGROUND AND SUMMARY OF THE INVENTION <EOH>The invention relates to a method and apparatus for outputting data providing information regarding a motor vehicle to a user of the motor vehicle.\",\n \"International Patent Document WO 98/34812 discloses a multimedia information and control system for a motor vehicle, in which the user selects the information of interest to him from a defined global set.\",\n \"Various options relating to the information are then offered, from which he can select at least one option and cause the respective data to be output.\",\n \"Furthermore, an operator control function which corresponds to the data that are output can be executed.\",\n \"One object of the invention is to provide an improved method and apparatus of the generic type in which operating convenience for the user is increased and operating safety and traffic safety are improved.\",\n \"This and other objects and advantages are achieved by the method and apparatus, according to the invention, in which the type and number of the options generated with respect to the information is determined and the respective data are output, as a function of at least one context-dependent profile, thus providing the user with fast and simple access to data which are more wide ranging in that they describe the individual information more closely.\",\n \"As used herein, the word “information” designates a generic classification or heading under which various data can be grouped, for example “windshield wiper”.\",\n \"The data relating to particular information comprises assigned details which are stored in a memory and which can be output in different ways for the user: in the case of “windshield wiper”, for example, an introduction video for operator control or a brief audio explanation of the interval levels.\",\n \"The context-dependent profile relates to the characterization of at least one secondary factor in the interaction between the vehicle, the vehicle user and the environment (for example the user's age).\",\n \"By reference to the context dependent profile, the options which are associated with the “information” (e.g., windshield wipers) are conditioned in such a way that from a total set of options, only those options which are assigned to the context-dependent profile are generated.\",\n \"Furthermore, by using the context-dependent profile it is also possible to output data automatically, without the user's previously having selected an option.\",\n \"By means of the context-dependent profile the number of generated options from which the user can make a selection and the outputting of the data are determined in such a manner that a clearly organized and user-compatible representation is made possible.\",\n \"Thus, when an item of information relating to the motor vehicle is requested, a subset of the entire set of data relating to this information is selected as a function of the profiles.\",\n \"(The subset can of course also correspond to the entire data set.)\",\n \"The selected data are output in a way which is particularly data-optimized for the user.\",\n \"As a result, the sequence of selecting data and outputting data is shortened, depending on the context.\",\n \"The invention also makes a contribution both to convenience and to safety.\",\n \"For example, when a vehicle display lights up, the user is automatically offered an explanation of this vehicle display as a function of the profile, by reason of appropriate data being output.\",\n \"The user is thus immediately informed, without having to think about the explanation or even perform awkward control operations.\",\n \"Likewise it possible to provide the user with selective proposals as a function of the profiles and to instruct him about vehicle functions which are suitable to use in a specific situation.\",\n \"It is advantageous if an operator control function whose use is proposed with the data which is output, is triggered as a function of the profiles and options.\",\n \"As a result, the operating convenience is increased further, because the user does not need to search for the triggering means which are assigned to this operator control function by virtue of the fact that the proposed operator control function is triggered in an easy way which is optimized as much as possible with respect to time.\",\n \"Such direct triggering of the operator control function, without previous interrogation of the user makes it possible to trigger the operator control function much more reliably in particular in driving situations for which timing is critical.\",\n \"Alternatively, direct triggering of the operator control function may be provided.\",\n \"In this case, as a function of the profiles, a corresponding option is defined, said option being selected and then bringing about indirect triggering of the operator control function.\",\n \"An example of direct or indirect triggering occurs if the driver selects the reverse gear on his vehicle, and as a result, either an electrically movable rear roller blind is retracted (direct triggering) or the retraction of the rear roller blind is proposed as an option (indirect triggering).\",\n \"One advantageous embodiment of the invention selects, for a particular user, a context-dependent profile based on the assignment of that user to a defined user group.\",\n \"As a result, convenience is increased further in that a specific, group related information requirement is fulfilled.\",\n \"For example, the assignment of the user to a user group takes into account whether he or she is already familiar with the vehicle or the make of vehicle.\",\n \"For example, a first user group may comprise users for which the vehicle or the make of vehicle is new, and a second user group may comprise experienced users.\",\n \"If, for example, a user is assigned to the first user group, a specific context dependent profile is defined, bringing about an additional, explanatory outputting of data, for example relating to operator control elements.\",\n \"Alternatively or additionally, a brief explanation on the vehicle and its operator control elements is possible when the user is assigned to the first user group.\",\n \"The described assignment to user groups is carried out, for example, during visits for servicing of the vehicle.\",\n \"Thus, when the vehicle is handed back the “new user” profile can then be defined.\",\n \"On one of the subsequent visits to the workshop it is then possible to define the “experienced users” group.\",\n \"A further possibility of a predefinable user group is to select an “interested user” profile which, for example with respect to predefinable vehicle reactions, supplies data in the form of explanations or technical explanations on vehicle functions.\",\n \"By assigning the user to a user group for the disabled it is possible to output data as a function of the profile in order to support the user in specific situations.\",\n \"For example, given an assignment of the user to a “restricted hearing” user group, as a function of the profiles, supplementary visual data are output if according to standard practice an operator control element provides audible feedback in the vehicle.\",\n \"A user can also be assigned to more than one user group, so that more than one profile is assigned for him, preferably with different priorities assigned to different profiles.\",\n \"If a signal for representing options and/or for outputting data (below for the sake of simplification we only refer to the outputting of options or data) from two or more profiles is present, the options or data are output in correct order and as a function of the priority of the profile.\",\n \"Here, for example, the options or data with the highest priority are output immediately and additionally an option which the user can select in order to cause the further options or data to be output is illustrated.\",\n \"It is also possible to define separate profiles for a plurality of users, for example for a user who drives the vehicle, a user who is a front seat passenger, or a user who is sitting in the rear of the vehicle.\",\n \"In a profile which is defined for the driver of the vehicle, functions relating, for example, to driving safety are provided, while in a profile defined for the rear-seat passengers, audio and multimedia functions are provided, for example.\",\n \"In this manner, the operating reliability and thus the traffic safety are increased by the context-dependent profile for the driver of the vehicle.\",\n \"Similarly, for a rear-seat passenger the operating convenience is increased, and comprehensive information about the operator control functions which are available is provided.\",\n \"In addition it is also thus possible to define further profiles, for example it is possible to provide a specific profile for a service technician with a representation of options and/or outputting of data on the diagnostics of the vehicle.\",\n \"A further advantageous embodiment of the invention selects a context-dependent profile for a predefinable vehicle environment.\",\n \"Here, a user is informed about changes which have occurred in the course of his journey by presenting options and/or data which permit the user to prepare for changed conditions on his route, as a function of the profiles.\",\n \"For example, it is possible for him to obtain information about operator control functions which are suitable for the new conditions and to trigger these functions.\",\n \"Thus, in the case of a road in poor condition which is stored as an attribute in the digital road map of the vehicle, it is possible to present options as a function of the profiles and/or to output data relating to the operator control elements of a ride level controller (which sets the ground clearance of the motor vehicle).\",\n \"Furthermore, it is possible, for example, to define a country-specific profile.\",\n \"When it is detected that a national border has been crossed, information about country-specific traffic regulations are provided as a function of the profiles.\",\n \"A conventional device for locating the position of the vehicle is used, for example, to identify the border crossing.\",\n \"In a further refinement of the invention, context dependent profiles are defined with respect to the type of vehicle.\",\n \"As a result, a user is informed about changes which occur when the type of vehicle which he uses at a particular time changes.\",\n \"If the user changes, for example, from his previous vehicle model of his make of vehicle to a newer model, only options or data which relate to operator control elements which have changed or are new in comparison to his previously used vehicle are issued to him.\",\n \"This applies in an analogous way, for example, to retrofits or software updates of vehicles.\",\n \"In each case the differences between the current vehicle and the vehicle which is known to the user is determined first, and only those options or data which are associated with this difference for the user are output to the user as a function of the profiles.\",\n \"As a result, by virtue of the profiles, the user is informed about the possibilities which the currently used vehicle provides.\",\n \"These context-dependent profiles are preferably defined for a user individually according to a type of vehicle.\",\n \"It is possible, for example, for a “first user” profile to be present which explains innovations only a single time, while a “second user” profile explains these changes within a specific time period whenever a journey is begun.\",\n \"It is very advantageous to define context dependent profiles which output options and/or data as a function of at least one current vehicle variable, such as for example, currently measured sensor data or vehicle system states such as the heating level of the seat heating system or the range of the vehicle with the currently available quantity of fuel which is determined by the trip computer.\",\n \"As a result, a vehicle user can be informed particularly quickly about specific situations because, data corresponding to the situation indicated by the vehicle variables is issued to him, as a function of the profiles.\",\n \"For example, options or data relating to a TCS (Traction Control System) can be output if the wheels spin during the process of driving off.\",\n \"It is also possible to output, as a function of the profiles, instructions on “cutting free” (by the user briefly switching off the TCS during the process of driving off), by virtue of the fact that the operator control element for switching off the TCS is pointed out and/or direct triggering—i.e.\",\n \"deactivation—with or without previous interrogation of the user is brought about.\",\n \"In the case of a flat tire or an accident it is possible, for example, to issue options or data on the correct behavior in emergency situations and/or on the position of the warning triangle in the vehicle.\",\n \"The vehicle variables can also be transmitted from outside the vehicle, for example in wireless fashion by XFCD (“Extended Floating Car Data”).\",\n \"It is thus possible to transmit into the vehicle sensor data about weather conditions on the route ahead.\",\n \"The user is then issued with options or data as a function of the profiles such that the user can set an air conditioning system in his vehicle in a way which is advantageous for these conditions.\",\n \"A further advantageous embodiment of the invention provides for a plausibility check for a control operation which is carried out by the user, with the options and/or the data being output as a function of the result of the plausibility check.\",\n \"This increases the operator control reliability and increases the convenience because a user is offered information about an operator control function or its operator control element as a function of the profile if he makes an operator control error or if an algorithm detects an operator control problem.\",\n \"As a result, it is possible, for example when there is multiple use of the operator control elements for main beam and/or indicator lights within a short period of time while it is raining at the same time, to offer and/or issue data on the operator control of the windshield wiper, in particular on how to switch it on, as options for selection.\",\n \"In a further configuration of the invention, vehicle variables and/or operator control operations are stored in a memory, which is initially interrogated before options are represented and/or data is output as a function of the profiles.\",\n \"If values relating to preceding vehicle variables or operator control operations are stored in the memory, these values are checked to determine whether they are to be taken into account in the representation of the current options and/or in the outputting of the current data.\",\n \"This further increases operating convenience because a rapid selection of options and/or data is matched individually to the user.\",\n \"As it were, a “memory” is provided by virtue of the fact that conclusions can be drawn relating to a current operator control operation, for example from analyzing preceding operator control operations.\",\n \"In the case of operator control steps which the user usually makes, it is thus possible to suppress a representation of information and outputting of data.\",\n \"The classification of operator control problems or the categorization of current vehicle variables is also improved by virtue of the fact that, for example, operator control steps which are specific to a user are detected and no longer classified as an operator control problem and explanatory data is output.\",\n \"Such a recognition can be achieved, for example, by using the methods of “artificial intelligence” (such as a neural network).\",\n \"By checking of values stored in the memory it is also possible to make modifications to the currently defined, context-dependent profile itself and/or retrieve and subsequently define a different context dependent profile.\",\n \"It is advantageous if at least part of an operating manual for the motor vehicle is represented as information relating to the motor vehicle.\",\n \"A method suitable for this purpose is described, for example, in the German Patent Application DE 199 59 755.3-21 whose content is included herein by reference.\",\n \"In this method, a current equipment variant of the motor vehicle is determined in a preceding step and a second database is generated from a first database as a function of this determined variant, this second database comprising only information on the operator control for the current equipment variant of the motor vehicle.\",\n \"In this way, a continuously updated information state relating to the motor vehicle and its operator control elements is ensured.\",\n \"The device according to the invention for processing and outputting data relating to information regarding a motor vehicle or its operator control elements comprises a display for representing options relating to the information (referred to below for short as first output means), a display for the user to select at least one option (referred to below for short as selection means), a display for outputting data relating to the selected option (referred to for short below as second output means) and a processing unit which generates, by reference to at least one defined, context-dependent profile, a number of options that are characterized by the profile and which processing unit outputs data as a function of the profiles and options.\",\n \"This processing unit may be embodied, for example, as a vehicle computer (a processing unit which is arranged in the vehicle).\",\n \"However, it can also be implemented by providing additional functions to a unit which is already present in the vehicle—for example a control device.\",\n \"The first output means and/or the second output means may be, for example, display units of conventional design such as LCD (Liquid Crystal Display), or may be presented on the windshield (Head Up Display).\",\n \"It is advantageous here to embody the display unit as a function of the set of characteristics of the options and/or data (both of these are referred to below as data set).\",\n \"For example, a small data set may be output by means of a display which is already present in the vehicle, while a relatively large data set is output by means of a further relatively large display which is mounted on the dashboard or in the center console.\",\n \"In addition, further possibilities for displaying options and/or data are available.\",\n \"The outputting of the options and data can also depend on what technical equipment a vehicle has.\",\n \"For example, when a suitable display fails, it is possible to suppress the outputting of certain data (for example video sequences forming a short introduction to the vehicle for a new user).\",\n \"Of course, it is also possible to provide a joint display for outputting the options and data.\",\n \"In order to integrate the first and/or second output means with the selection means, a conventional touch-sensitive display (touch screen) is preferably provided.\",\n \"Alternatively or additionally, the selection means may be embodied as a voice operated controller, such as is already used today in motor vehicles, permitting the driver to concentrate his visual attention on the events on the road.\",\n \"Furthermore, the selection means may be embodied as a manual operator control element of conventional design, for example, in the form of a joystick, or as a combined rotary/pushbutton knob which has in each case a different number of latching positions depending on the number of options for the information.\",\n \"Such an operator control element is also already present in many vehicles (for example for controlling a navigation device), and can be used advantageously.\",\n \"Further possible operator control systems may be provided.\",\n \"In order to output the options or the selected data it is possible to provide means of a conventional design, for example audible outputs, such as an audio system, or haptic output (for example a force feedback operator control element, an opposing force being exerted at said operator control element when it is used).\",\n \"The means which is used for inputting and/or outputting may also be defined in a variable fashion as a function of the profiles, for example a visual output is replaced by an audible output as soon as the vehicle exceeds a defined limiting speed.\",\n \"Furthermore, the data which are output can be further processed and fed to other systems, in particular assistance systems in a motor vehicle.\",\n \"Other objects, advantages and novel features of the present invention will become apparent from the following detailed description of the invention when considered in conjunction with the accompanying drawings.\"\n ],\n [\n \"This application claims the priority of German patent document 101 17 410.1, filed Apr.\",\n \"6, 2001 (PCT No.\",\n \"PCT/EP02/01485) filed Feb. 13, 2002), the disclosure of which is expressly incorporated by reference herein.\",\n \"BACKGROUND AND SUMMARY OF THE INVENTION The invention relates to a method and apparatus for outputting data providing information regarding a motor vehicle to a user of the motor vehicle.\",\n \"International Patent Document WO 98/34812 discloses a multimedia information and control system for a motor vehicle, in which the user selects the information of interest to him from a defined global set.\",\n \"Various options relating to the information are then offered, from which he can select at least one option and cause the respective data to be output.\",\n \"Furthermore, an operator control function which corresponds to the data that are output can be executed.\",\n \"One object of the invention is to provide an improved method and apparatus of the generic type in which operating convenience for the user is increased and operating safety and traffic safety are improved.\",\n \"This and other objects and advantages are achieved by the method and apparatus, according to the invention, in which the type and number of the options generated with respect to the information is determined and the respective data are output, as a function of at least one context-dependent profile, thus providing the user with fast and simple access to data which are more wide ranging in that they describe the individual information more closely.\",\n \"As used herein, the word “information” designates a generic classification or heading under which various data can be grouped, for example “windshield wiper”.\",\n \"The data relating to particular information comprises assigned details which are stored in a memory and which can be output in different ways for the user: in the case of “windshield wiper”, for example, an introduction video for operator control or a brief audio explanation of the interval levels.\",\n \"The context-dependent profile relates to the characterization of at least one secondary factor in the interaction between the vehicle, the vehicle user and the environment (for example the user's age).\",\n \"By reference to the context dependent profile, the options which are associated with the “information” (e.g., windshield wipers) are conditioned in such a way that from a total set of options, only those options which are assigned to the context-dependent profile are generated.\",\n \"Furthermore, by using the context-dependent profile it is also possible to output data automatically, without the user's previously having selected an option.\",\n \"By means of the context-dependent profile the number of generated options from which the user can make a selection and the outputting of the data are determined in such a manner that a clearly organized and user-compatible representation is made possible.\",\n \"Thus, when an item of information relating to the motor vehicle is requested, a subset of the entire set of data relating to this information is selected as a function of the profiles.\",\n \"(The subset can of course also correspond to the entire data set.)\",\n \"The selected data are output in a way which is particularly data-optimized for the user.\",\n \"As a result, the sequence of selecting data and outputting data is shortened, depending on the context.\",\n \"The invention also makes a contribution both to convenience and to safety.\",\n \"For example, when a vehicle display lights up, the user is automatically offered an explanation of this vehicle display as a function of the profile, by reason of appropriate data being output.\",\n \"The user is thus immediately informed, without having to think about the explanation or even perform awkward control operations.\",\n \"Likewise it possible to provide the user with selective proposals as a function of the profiles and to instruct him about vehicle functions which are suitable to use in a specific situation.\",\n \"It is advantageous if an operator control function whose use is proposed with the data which is output, is triggered as a function of the profiles and options.\",\n \"As a result, the operating convenience is increased further, because the user does not need to search for the triggering means which are assigned to this operator control function by virtue of the fact that the proposed operator control function is triggered in an easy way which is optimized as much as possible with respect to time.\",\n \"Such direct triggering of the operator control function, without previous interrogation of the user makes it possible to trigger the operator control function much more reliably in particular in driving situations for which timing is critical.\",\n \"Alternatively, direct triggering of the operator control function may be provided.\",\n \"In this case, as a function of the profiles, a corresponding option is defined, said option being selected and then bringing about indirect triggering of the operator control function.\",\n \"An example of direct or indirect triggering occurs if the driver selects the reverse gear on his vehicle, and as a result, either an electrically movable rear roller blind is retracted (direct triggering) or the retraction of the rear roller blind is proposed as an option (indirect triggering).\",\n \"One advantageous embodiment of the invention selects, for a particular user, a context-dependent profile based on the assignment of that user to a defined user group.\",\n \"As a result, convenience is increased further in that a specific, group related information requirement is fulfilled.\",\n \"For example, the assignment of the user to a user group takes into account whether he or she is already familiar with the vehicle or the make of vehicle.\",\n \"For example, a first user group may comprise users for which the vehicle or the make of vehicle is new, and a second user group may comprise experienced users.\",\n \"If, for example, a user is assigned to the first user group, a specific context dependent profile is defined, bringing about an additional, explanatory outputting of data, for example relating to operator control elements.\",\n \"Alternatively or additionally, a brief explanation on the vehicle and its operator control elements is possible when the user is assigned to the first user group.\",\n \"The described assignment to user groups is carried out, for example, during visits for servicing of the vehicle.\",\n \"Thus, when the vehicle is handed back the “new user” profile can then be defined.\",\n \"On one of the subsequent visits to the workshop it is then possible to define the “experienced users” group.\",\n \"A further possibility of a predefinable user group is to select an “interested user” profile which, for example with respect to predefinable vehicle reactions, supplies data in the form of explanations or technical explanations on vehicle functions.\",\n \"By assigning the user to a user group for the disabled it is possible to output data as a function of the profile in order to support the user in specific situations.\",\n \"For example, given an assignment of the user to a “restricted hearing” user group, as a function of the profiles, supplementary visual data are output if according to standard practice an operator control element provides audible feedback in the vehicle.\",\n \"A user can also be assigned to more than one user group, so that more than one profile is assigned for him, preferably with different priorities assigned to different profiles.\",\n \"If a signal for representing options and/or for outputting data (below for the sake of simplification we only refer to the outputting of options or data) from two or more profiles is present, the options or data are output in correct order and as a function of the priority of the profile.\",\n \"Here, for example, the options or data with the highest priority are output immediately and additionally an option which the user can select in order to cause the further options or data to be output is illustrated.\",\n \"It is also possible to define separate profiles for a plurality of users, for example for a user who drives the vehicle, a user who is a front seat passenger, or a user who is sitting in the rear of the vehicle.\",\n \"In a profile which is defined for the driver of the vehicle, functions relating, for example, to driving safety are provided, while in a profile defined for the rear-seat passengers, audio and multimedia functions are provided, for example.\",\n \"In this manner, the operating reliability and thus the traffic safety are increased by the context-dependent profile for the driver of the vehicle.\",\n \"Similarly, for a rear-seat passenger the operating convenience is increased, and comprehensive information about the operator control functions which are available is provided.\",\n \"In addition it is also thus possible to define further profiles, for example it is possible to provide a specific profile for a service technician with a representation of options and/or outputting of data on the diagnostics of the vehicle.\",\n \"A further advantageous embodiment of the invention selects a context-dependent profile for a predefinable vehicle environment.\",\n \"Here, a user is informed about changes which have occurred in the course of his journey by presenting options and/or data which permit the user to prepare for changed conditions on his route, as a function of the profiles.\",\n \"For example, it is possible for him to obtain information about operator control functions which are suitable for the new conditions and to trigger these functions.\",\n \"Thus, in the case of a road in poor condition which is stored as an attribute in the digital road map of the vehicle, it is possible to present options as a function of the profiles and/or to output data relating to the operator control elements of a ride level controller (which sets the ground clearance of the motor vehicle).\",\n \"Furthermore, it is possible, for example, to define a country-specific profile.\",\n \"When it is detected that a national border has been crossed, information about country-specific traffic regulations are provided as a function of the profiles.\",\n \"A conventional device for locating the position of the vehicle is used, for example, to identify the border crossing.\",\n \"In a further refinement of the invention, context dependent profiles are defined with respect to the type of vehicle.\",\n \"As a result, a user is informed about changes which occur when the type of vehicle which he uses at a particular time changes.\",\n \"If the user changes, for example, from his previous vehicle model of his make of vehicle to a newer model, only options or data which relate to operator control elements which have changed or are new in comparison to his previously used vehicle are issued to him.\",\n \"This applies in an analogous way, for example, to retrofits or software updates of vehicles.\",\n \"In each case the differences between the current vehicle and the vehicle which is known to the user is determined first, and only those options or data which are associated with this difference for the user are output to the user as a function of the profiles.\",\n \"As a result, by virtue of the profiles, the user is informed about the possibilities which the currently used vehicle provides.\",\n \"These context-dependent profiles are preferably defined for a user individually according to a type of vehicle.\",\n \"It is possible, for example, for a “first user” profile to be present which explains innovations only a single time, while a “second user” profile explains these changes within a specific time period whenever a journey is begun.\",\n \"It is very advantageous to define context dependent profiles which output options and/or data as a function of at least one current vehicle variable, such as for example, currently measured sensor data or vehicle system states such as the heating level of the seat heating system or the range of the vehicle with the currently available quantity of fuel which is determined by the trip computer.\",\n \"As a result, a vehicle user can be informed particularly quickly about specific situations because, data corresponding to the situation indicated by the vehicle variables is issued to him, as a function of the profiles.\",\n \"For example, options or data relating to a TCS (Traction Control System) can be output if the wheels spin during the process of driving off.\",\n \"It is also possible to output, as a function of the profiles, instructions on “cutting free” (by the user briefly switching off the TCS during the process of driving off), by virtue of the fact that the operator control element for switching off the TCS is pointed out and/or direct triggering—i.e.\",\n \"deactivation—with or without previous interrogation of the user is brought about.\",\n \"In the case of a flat tire or an accident it is possible, for example, to issue options or data on the correct behavior in emergency situations and/or on the position of the warning triangle in the vehicle.\",\n \"The vehicle variables can also be transmitted from outside the vehicle, for example in wireless fashion by XFCD (“Extended Floating Car Data”).\",\n \"It is thus possible to transmit into the vehicle sensor data about weather conditions on the route ahead.\",\n \"The user is then issued with options or data as a function of the profiles such that the user can set an air conditioning system in his vehicle in a way which is advantageous for these conditions.\",\n \"A further advantageous embodiment of the invention provides for a plausibility check for a control operation which is carried out by the user, with the options and/or the data being output as a function of the result of the plausibility check.\",\n \"This increases the operator control reliability and increases the convenience because a user is offered information about an operator control function or its operator control element as a function of the profile if he makes an operator control error or if an algorithm detects an operator control problem.\",\n \"As a result, it is possible, for example when there is multiple use of the operator control elements for main beam and/or indicator lights within a short period of time while it is raining at the same time, to offer and/or issue data on the operator control of the windshield wiper, in particular on how to switch it on, as options for selection.\",\n \"In a further configuration of the invention, vehicle variables and/or operator control operations are stored in a memory, which is initially interrogated before options are represented and/or data is output as a function of the profiles.\",\n \"If values relating to preceding vehicle variables or operator control operations are stored in the memory, these values are checked to determine whether they are to be taken into account in the representation of the current options and/or in the outputting of the current data.\",\n \"This further increases operating convenience because a rapid selection of options and/or data is matched individually to the user.\",\n \"As it were, a “memory” is provided by virtue of the fact that conclusions can be drawn relating to a current operator control operation, for example from analyzing preceding operator control operations.\",\n \"In the case of operator control steps which the user usually makes, it is thus possible to suppress a representation of information and outputting of data.\",\n \"The classification of operator control problems or the categorization of current vehicle variables is also improved by virtue of the fact that, for example, operator control steps which are specific to a user are detected and no longer classified as an operator control problem and explanatory data is output.\",\n \"Such a recognition can be achieved, for example, by using the methods of “artificial intelligence” (such as a neural network).\",\n \"By checking of values stored in the memory it is also possible to make modifications to the currently defined, context-dependent profile itself and/or retrieve and subsequently define a different context dependent profile.\",\n \"It is advantageous if at least part of an operating manual for the motor vehicle is represented as information relating to the motor vehicle.\",\n \"A method suitable for this purpose is described, for example, in the German Patent Application DE 199 59 755.3-21 whose content is included herein by reference.\",\n \"In this method, a current equipment variant of the motor vehicle is determined in a preceding step and a second database is generated from a first database as a function of this determined variant, this second database comprising only information on the operator control for the current equipment variant of the motor vehicle.\",\n \"In this way, a continuously updated information state relating to the motor vehicle and its operator control elements is ensured.\",\n \"The device according to the invention for processing and outputting data relating to information regarding a motor vehicle or its operator control elements comprises a display for representing options relating to the information (referred to below for short as first output means), a display for the user to select at least one option (referred to below for short as selection means), a display for outputting data relating to the selected option (referred to for short below as second output means) and a processing unit which generates, by reference to at least one defined, context-dependent profile, a number of options that are characterized by the profile and which processing unit outputs data as a function of the profiles and options.\",\n \"This processing unit may be embodied, for example, as a vehicle computer (a processing unit which is arranged in the vehicle).\",\n \"However, it can also be implemented by providing additional functions to a unit which is already present in the vehicle—for example a control device.\",\n \"The first output means and/or the second output means may be, for example, display units of conventional design such as LCD (Liquid Crystal Display), or may be presented on the windshield (Head Up Display).\",\n \"It is advantageous here to embody the display unit as a function of the set of characteristics of the options and/or data (both of these are referred to below as data set).\",\n \"For example, a small data set may be output by means of a display which is already present in the vehicle, while a relatively large data set is output by means of a further relatively large display which is mounted on the dashboard or in the center console.\",\n \"In addition, further possibilities for displaying options and/or data are available.\",\n \"The outputting of the options and data can also depend on what technical equipment a vehicle has.\",\n \"For example, when a suitable display fails, it is possible to suppress the outputting of certain data (for example video sequences forming a short introduction to the vehicle for a new user).\",\n \"Of course, it is also possible to provide a joint display for outputting the options and data.\",\n \"In order to integrate the first and/or second output means with the selection means, a conventional touch-sensitive display (touch screen) is preferably provided.\",\n \"Alternatively or additionally, the selection means may be embodied as a voice operated controller, such as is already used today in motor vehicles, permitting the driver to concentrate his visual attention on the events on the road.\",\n \"Furthermore, the selection means may be embodied as a manual operator control element of conventional design, for example, in the form of a joystick, or as a combined rotary/pushbutton knob which has in each case a different number of latching positions depending on the number of options for the information.\",\n \"Such an operator control element is also already present in many vehicles (for example for controlling a navigation device), and can be used advantageously.\",\n \"Further possible operator control systems may be provided.\",\n \"In order to output the options or the selected data it is possible to provide means of a conventional design, for example audible outputs, such as an audio system, or haptic output (for example a force feedback operator control element, an opposing force being exerted at said operator control element when it is used).\",\n \"The means which is used for inputting and/or outputting may also be defined in a variable fashion as a function of the profiles, for example a visual output is replaced by an audible output as soon as the vehicle exceeds a defined limiting speed.\",\n \"Furthermore, the data which are output can be further processed and fed to other systems, in particular assistance systems in a motor vehicle.\",\n \"Other objects, advantages and novel features of the present invention will become apparent from the following detailed description of the invention when considered in conjunction with the accompanying drawings.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS FIG.\",\n \"1 shows a schematic block circuit diagram of an embodiment of the device for outputting data relating to information regarding a motor vehicle; FIG.\",\n \"2 shows a schematic block circuit diagram of a further embodiment of the device; FIG.\",\n \"3 shows a plan view of an embodiment of the device; FIG.\",\n \"4 shows, by way of example, an image which is represented on a display unit, and is generated by the device in a basic state; FIG.\",\n \"5 shows, by way of example, an image which is represented on a display unit, and is generated by the device in a state with a preselected option; and FIG.\",\n \"6 shows, by way of example, an image which is represented on a display unit, and is generated by the device in a state with output data.\",\n \"DETAILED DESCRIPTION OF THE DRAWINGS FIG.\",\n \"1 shows a motor vehicle 1 with a device 2 for outputting data (D) with a processing unit 3.The device 2 comprises a means 4 for representing options (On) relating to the information (I) (referred to below for short as options output means 4), and a means 5 for outputting data (D) relating to the selected option (O) (referred to below for short as data output means 5).\",\n \"The two output means 4 and 5 are, for example, two displays which are arranged at a distance from one another.\",\n \"Alternatively, both output means 4 and 5 can be formed by a display.\",\n \"As a further alternative or additionally, an audible output is provided for the output means 4 and/or 5.Furthermore, the processing unit 3 has a means 6 (referred to below for short as selection means 6) by which the user can select at least one option (O).\",\n \"The selection means 6 is embodied as a separate unit, for example, as a voice operated controller.\",\n \"Additionally or alternatively, a manual operator control element of conventional design may be provided, such as for example a joystick or a combined rotary/pushbutton knob, which has a large number of different latched positions depending on the number of options (On) for the information.\",\n \"As a further alternative it is possible to provide a unit which detects body movements or facial expression of the user, classified them and assigns them to a corresponding operator control request, for which the requested operator control operation is then carried out.\",\n \"The processing unit 3 is supplied with values (W) from at least one component 7, such as a vehicle-mounted sensor which measures the current speed, or a control unit which is provided for setting possibilities of the vehicle seat.\",\n \"However, it is also possible to transmit values in a wireless fashion into the vehicle, for example from other vehicles and/or from local transmission devices arranged at the edge of the road pavement.\",\n \"Furthermore, a unit 8 for storing values (Wn) such as, for example, sensor data and/or operator control operations is provided (referred to below as storage unit 8).\",\n \"The storage unit 8 may be embodied as part of the usual peripheral components of the processing unit 3 with a conventional design.\",\n \"Furthermore, at least one context-dependent profile (P) is selected from a memory 9 by at least one profile selection means 10, and fed to the processing unit 3.The profile selection means 10 may provide a switch, for example a rotary or pushbutton switch, which is suitable for this.\",\n \"The user places the switch in a switch position which is provided for it and which is sensed by the processing unit 3, after which it retrieves the context-dependent profile or profiles (P) which is selected by the user from the memory 9 and allocates them to the user.\",\n \"Alternatively, the profile selection means 10 is connected to the starting device of the engine as a fingerprint sensor (detection of the “fingerprint” of the user).\",\n \"When the user initiates an engine starting procedure, he is identified by the profile selection means 10.Then, at least one context dependent profile (P) which is provided for the user is retrieved from the memory 9 and defined for him.\",\n \"Likewise, it is possible to provide a specific interface of the processor unit 3 as the profile selection means 10, to which interface, for example, a customary diagnostic device for defining a context dependent profile (P) can be connected.\",\n \"Alternatively or additionally, an electronic data carrier can be provided for the combination of memory 9 and profile selection means 10, the user carrying said data carrier, for example a driver-specific chip card, on him and introducing it into a device which is connected to the vehicle.\",\n \"The user-specific values which are evaluated by the processing unit 3 and bring about the selection of a context-dependent profile (P) which is suitable for the user are stored on the chip card.\",\n \"The electronic data carrier can also be used for storing other values.\",\n \"For example, the data carrier can also be provided for storing values (Wn) such as, for example, sensor data and/or operator control operations of the user.\",\n \"Alternatively or additionally, the memory 9 can also be provided outside the vehicle for example, in a service center (a central processing unit with a database which can be used in a wireless fashion by subscribers to a respective service).\",\n \"A context-dependent profile (P) which is provided for the user is transferred from the service center into the vehicle in a wireless fashion if it is detected in this control center that a transceiver unit (“radio key”) which is carried by the user initiates vehicle starting procedure.\",\n \"Alternatively or additionally, the service center is also used to store vehicle-related values (Wn) such as, for example, sensor data and/or operator control operations of the user.\",\n \"The selection of the information (I), the type and number of options (On) generated with respect to the information (I) and the outputting of the respective data (D) are controlled by the context-dependent profile (P) by means of the processing unit 3.The data (D) which are to be output are selected from a database 11 as a function of profiles and/or options, read and output using the output means 5.Depending on the type and design of the device 2, the database 11 is integrated in the motor vehicle 1.Alternatively or additionally, data from an external database 12 can be read by the processing device 2 via wireless transmission R. This external database 12 can be provided in the service center described above.\",\n \"FIG.\",\n \"2 illustrates a further embodiment of the device 2 for outputting data (D) relating to the information (I).\",\n \"The device 2 comprises two separate networks 13, 14, which are embodied as units of network nodes that are each connected to a plurality of transmission lines.\",\n \"This distributed design divides the functional units of the device 2 (shown in FIG.\",\n \"1) into the network 13 in which the values (W) of the component 7 are collected and conditioned, while storage, conditioning and outputting of data (D) is carried out by the network 14.The networks 13, 14 are connected to one another by a transmission line 15.In order to connect the two networks 13, 14 to the transmission line 15, the gateway network node 16 is provided in the network 13 and the gateway network node 17 is provided in the network 14.As is schematically illustrated in FIG.\",\n \"2, the two networks 13, 14 have different architectures and are thus configured for different requirements.\",\n \"The network 13 has a tree structure and comprises components 7, the storage unit 8, the memory 9 and an associated display unit 18, while the network 14, which has a ring structure and is suitable particularly for transmitting large amounts of data, comprises the database 11 and an associated, large-format display unit 19.FIG.\",\n \"3 is a plan view of an embodiment of the device 2 in the dashboard of the motor vehicle 1, including a display unit 18, the selection means 6 for selecting options (On) and the display unit 19.The processing unit 3 causes data (D) relating to information (I) to be represented on the display unit 18, as a function of profiles and/or options.\",\n \"At the same time, the information (I) designates a global term which can include various types of data (D).\",\n \"As is illustrated in block 20, explanations of warning displays and changes of states of the motor vehicle 1 as well as of exceptional situations and suitable operator control functions are output on the display unit 18 by the processing unit 3.When there is a more wide ranging information request relating to data (D) which has already been output in the display unit 18 in the form of brief information, further data (D) is output as a detailed explanation in the display unit 19 as a function of the profiles.\",\n \"The display units 18, 19 are appropriately embodied in terms of their size, type and arrangement in this respect.\",\n \"Additionally or alternatively, options (O) relating to the information (I) for the outputting of data are represented on the display unit 18, as illustrated by way of example in FIG.\",\n \"4.A respective option (Os) for the outputting of the data (D) which describe the respective information is selected by operating the selection means 6.When there is a direct request for information (I) by the user via the manual operated control element 6, corresponding data (D) are output on the display unit 19.This data comprise, as shown in block 22, an online help, a keyword/subject search feature, an initial instruction feature and an explanation of operator control switches, as shown in FIG.\",\n \"3.FIG.\",\n \"4 shows by way of example an image which is represented on the output means 4 or 5 and generated by the device 2 as a function of the profiles when the device 2 is in a basic state.\",\n \"In this state, the user can request data (D) directly as a function of the profiles.\",\n \"As shown in FIG.\",\n \"4 as a function of the context dependent profile, the user is provided, with the options 01 to 07 relating to the information (I) “motor vehicle information system”.\",\n \"The user can retrieve explanatory data (D) and/or further options (O) relating to the multipurpose instrument and to the central unit by selecting corresponding options 01, 02 by using the selection means 6.Here, a further option 08, 09 relating to the two first options 01, 02, which can be selected by selection, is already defined.\",\n \"The option 01 is already selected as a function of the profiles.\",\n \"Furthermore, the user can selectively search for subjects on which he desires further explanations by selecting the option 03 “subject search”.\",\n \"If the user selects the option 04 “explanation of switch”, he can cause data (D) and/or further options (O) relating to individual operator control elements to be output directly.\",\n \"The selection of the option 05 “initial instruction” by the user causes data (D) and/or further options (O) to be output which are advantageous for the user if he is using the motor vehicle for the first time.\",\n \"By selecting the option 06 “last subject” data which the user had already requested once at an earlier time is output to the user.\",\n \"Selecting the option 07 “settings” provides the user with information on the device according to the invention itself.\",\n \"FIG.\",\n \"5 shows by way of example a further image which is represented on the output means 4 or 5.The image represents to the user that the device 2 is in a state in which an option (O) has already been selected.\",\n \"In this state, the user can cause the corresponding data (D) to be output automatically as a function of the profiles, and as a function of options by selecting the respective option (O).\",\n \"Likewise, a number of symbols A indicate to the user how many options 0 have already been selected.\",\n \"Here, the respectively generated images are depicted one on top of the other as a hierarchical structure.\",\n \"The direct representation of such an image as a function of the profiles according to the invention avoids the tedious selection process which would otherwise precede a representation of this image.\",\n \"Directly selecting a symbol A causes the respective image to be represented directly in the hierarchical structure.\",\n \"A symbol (W) indicates to the user that further options (O) have been generated, and these can be “scrolled through” by selecting the symbol (W).\",\n \"FIG.\",\n \"6 shows by way of example an image which is represented on the display unit 5.This image is generated by the device 2 if it is in a state with data (D) which have been output.\",\n \"As is apparent in FIG.\",\n \"6, these data (D) comprise a pictogram (P), which has been obtained from the operating manual of the motor vehicle 1, and explanatory text.\",\n \"Selecting the option 011 causes further data (D) to be output.\",\n \"Selecting the option 012 causes thematically related data (D) to be represented as options (O) for outputting.\",\n \"Selecting the option 013 causes the operator control function which is explained by the data (D) to be triggered.\",\n \"Selecting the option 014 causes the image generated by a preceding processing step to be represented.\",\n \"The foregoing disclosure has been set forth merely to illustrate the invention and is not intended to be limiting.\",\n \"Since modifications of the disclosed embodiments incorporating the spirit and substance of the invention may occur to persons skilled in the art, the invention should be construed to include everything within the scope of the appended claims and equivalents thereof.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10474189"}}},{"rowIdx":788,"cells":{"text":{"kind":"list like","value":[["Device for the production of an extruded profile","A food product comprising a sugar wafer preferably cone-shaped having a filling of a substantially water-free fat-based confectionery material and a process for preparing it which comprises filling the sugar wafer with the substantially water-free fat-based confectionery in a molten, semi-liquid or semi-solid mass, and allowing the filling to harden."],["1.A device for the production of an extruded profile by co-extruding differing materials, particularly an extruded profile, with at least three parallel and spaced bars, comprising at least two extruders that release the differing materials as well as flow channels (14, 16, 18, 20, 22, 24) leading to a tool that comprises outlet openings releasing the materials such as slots (26, 28, 30), wherein the tool comprises at least three elongated, one inner and two first outer, outlet slots (26, 28, 30), whereby the outer outlet slots (26, 28) are each connected to a first flow channel (16, 18) extending from a first junction point (10) that is connected to one of the extruders for the supply of a first material and the inner outlet opening (30) is connected to two second flow channels (22, 24) for the supply of a second material, between which one of the first flow channels (18) runs and which extend from a joint second junction point (12) that is connected to the second extruder.","2.A device according to claim 1, wherein the inner outlet opening such as outlet slot (30) and the outer outlet openings such as outlet slots (26, 28) run parallel to each other.","3.A device according to claim 1, wherein from the first junction point (10) further flow channels (14, 20) extend, which lead to additional preferably slot-shaped outlet openings (32, 34), which run in the transverse peripheral region of at least one of the first outer outlet openings such as longitudinal slots (26, 28).","4.A device according to claim 1, wherein the two second flow channels (22, 24), which surround one of the first flow channels (18), and the first flow channel run perpendicular or substantially perpendicular to each other in the overlapping area.","5.A device according to claim 1, wherein at least four flow channels (14, 16, 18, 20) extend from the first junction point (10), of which two join into the outer elongated first outlet openings (26, 28) that are arranged parallel to each other and the remaining join into additional outlet openings (32, 34) that run perpendicular in them, of which one, respectively, runs along the transverse peripheral region of the outer first outlet openings (26, 28).","6.A device according to claim 1, wherein from the first junction point (10) a flow channel (18) extends; which joins both into one of the outer first outlet openings (28) and in additional outlet openings (40, 42, 44, 46) that run on the outside along its longitudinal edge."],["The invention relates to a device for the production of an extruded profile, by extrusion of differing materials, particularly for the production of an extruded profile with at least three parallel and spaced bars, comprising at least two extruders that release the differing materials as well as flow channels leading to a tool that comprises outlet openings releasing the materials such as slots.","In order to be able to produce semi-finished products or profile consisting of plastic continuously, extrusion processes are used.","Here thermoplastic resins in the form of e.g.","powder or granules are placed in an extruder, mixed with stabilizers, slip additives and possibly coloring and filling agents and are then conveyed to a profile-determining extrusion tool by means of a screw.","During this process the materials are compressed, degassed and homogenized while simultaneously being mixed and kneaded.","This creates a homogeneous, flowing mixture, the so-called molding compound, which is heated to or kept at a temperature that is characterized in the lower range by plastification and in the upper range by decomposition of the material.","After traveling through an extrusion die specifying the geometry, the material runs through a calibration unit, which consists of a dry-wet calibration combination.","Subsequent to that, the material is guided through a full bath vacuum or spray vacuum tank with integrated calibrating cover.","A corresponding device is revealed in DE 197 44 515.Here the flow channels are united directly in front of a die front plate of the tool in order to produce an extruded profile.","From EP 0 299 736 A1 we know of a co-extrusion tool in which also several flow channels run, which are united within the tool in order to produce a multi-layer extruded profile, which exhibits a laminate design.","It is the object of the present invention to further develop a device of the afore-mentioned kind such that extruded profiles can be extruded, which can consist of differing material in order to be able to process in particular inexpensive, such as recycled, materials.","Hereby it shall be ensured that the material that is supposed to be extruded flows to the necessary extent to relatively wide slots that are arranged parallel to one another.","Pursuant to the invention the problem is essentially resolved in that the tool comprises at least three elongated, i.e.","one inner and two outer, outlet slots, wherein the outer outlet slots each are connected to a first flow channel, for the supply of a first material, extending from a first junction point, and the inner outlet opening is connected to two second flow channels, for the supply of a second material, between which one of the first flow channels runs and which extend from one joint second junction point.","Hereby the inner outlet slot and the outer outlet slots run parallel to each other.","The junction points as such are connected to an extruder, respectively.","To be able to produce even complex extruded profiles, one further development of the invention provides that further flow channels extend from the first junction point, which lead to additional preferably slot-shaped outlet openings running in the transverse peripheral region of at least one of the outer elongated outlet openings.","In particular, at least one additional flow channel extends from the first junction point, wherein said channel ends up at another outlet opening that runs in a transverse peripheral region of one of the elongated outlet openings, in particular in a transverse peripheral region of the inner outlet opening.","In a preferred embodiment, at least four flow channels extend from the first junction point, wherein two of said four channels lead into the outer elongated outlet openings and two lead perpendicular to them, of which one, respectively, extends along the transverse peripheral regions of the outer elongated outlet openings.","The outlet openings as such join at a distance from each other into an outlet die of the tool so that the desired geometry of the profile can be achieved.","Hereby inner rods, in particular the one extruded with the inner elongated outlet opening, can consist of less expensive resin such as recycled material.","Further details, benefits and features of the invention are revealed not only in the claims, the characteristics disclosed therein—either alone and/or in combination—but also in the following description of an exemplary embodiment shown in the drawing.","To produce profiles by means of extrusion, with said profiles consisting of sections comprising differing materials and relatively elongated bars that run parallel to each other in the range of for example 100 mm, pursuant to the invention a device with a tool is suggested, where the flow channels have the necessary diameter to guarantee material supply to outlet openings such as slots or sheet dies.","The only FIGURE shows in purely principle form in a diagrammatic view flow channels that run in the tool of an extrusion device, whereby the channels extend from one of a total of two junction points 10, 12, which each are connected to an extruder, via which the flow channels are supplied with the flowing molding compound.","In the example, four flow channels 14, 16, 18 and 20 extend from the first junction point 10 and two flow channels 22, 24 from the second junction point 12.As the basic diagram shows, the tool comprises three elongated outlet slots 26, 28, 30, which run parallel to each other, wherein the outer outlet slots 26, 28, which transition into corresponding outlet dies of the extrusion tool, are connected to the flow channels 16, 18 leading to the first junction point 10 and the inner outlet slot 30 is connected to the second junction point 12 via the flow channels 22, 24.In order to supply the outlet slot 30 to the necessary extent with flowing homogenized and compressed material, the outlet slot 30 is supplied via the two flow channels 22, 24 by the junction point 12, wherein the flow channels 22, 24 surround in some sections the flow channel 18 that leads to the upper outlet slot 30.In other words, the flow channels 22, 24 extend perpendicular or substantially perpendicular in the overlapping area to the flow channel 18.Due to the fact that the inner outlet slot 30 is connected to two flow channels 22, 24, sufficient molding compound can be supplied regardless of the small diameter of the flow channels 22, 24.In other words, due to the arrangement of the flow channels pursuant to the invention, the outlet slots 26, 28, 30 can be sufficiently supplied with the compressed flowing material.","Furthermore, apart from the flow channels 16, 18 that lead to the outer elongated slotted dies 26, 28, two additional flow channels 14, 20 extend from the first junction point 10, which join into slots 32, 34 that run perpendicular to the outlet slots 26, 28, 30 and along their transverse edges.","Moreover additional outlet openings 36, 38 or 40, 42, 44, 46 with a square or rectangular cross-section branch off the flow channels 14, 20 as well as off the flow channel 18 in order to be able to create the desired extruded profile.","Hereby outlet openings 40, 42, 44, 46 are arranged along the outer longitudinal side of the upper outlet slot 28, i.e.","on the side away from the inner slot 30.Due to the fact that the flow channels that extend from the second junction point 12 in some areas surround a flow channel 18 that extends from the first junction point 12, sufficient compressed flowing material can flow to the outlet slots 30 with optimal space utilization, whereby the junction points 10, 12 are connected to extruders by means of which the differing materials are plasticized."]],"string":"[\n [\n \"Device for the production of an extruded profile\",\n \"A food product comprising a sugar wafer preferably cone-shaped having a filling of a substantially water-free fat-based confectionery material and a process for preparing it which comprises filling the sugar wafer with the substantially water-free fat-based confectionery in a molten, semi-liquid or semi-solid mass, and allowing the filling to harden.\"\n ],\n [\n \"1.A device for the production of an extruded profile by co-extruding differing materials, particularly an extruded profile, with at least three parallel and spaced bars, comprising at least two extruders that release the differing materials as well as flow channels (14, 16, 18, 20, 22, 24) leading to a tool that comprises outlet openings releasing the materials such as slots (26, 28, 30), wherein the tool comprises at least three elongated, one inner and two first outer, outlet slots (26, 28, 30), whereby the outer outlet slots (26, 28) are each connected to a first flow channel (16, 18) extending from a first junction point (10) that is connected to one of the extruders for the supply of a first material and the inner outlet opening (30) is connected to two second flow channels (22, 24) for the supply of a second material, between which one of the first flow channels (18) runs and which extend from a joint second junction point (12) that is connected to the second extruder.\",\n \"2.A device according to claim 1, wherein the inner outlet opening such as outlet slot (30) and the outer outlet openings such as outlet slots (26, 28) run parallel to each other.\",\n \"3.A device according to claim 1, wherein from the first junction point (10) further flow channels (14, 20) extend, which lead to additional preferably slot-shaped outlet openings (32, 34), which run in the transverse peripheral region of at least one of the first outer outlet openings such as longitudinal slots (26, 28).\",\n \"4.A device according to claim 1, wherein the two second flow channels (22, 24), which surround one of the first flow channels (18), and the first flow channel run perpendicular or substantially perpendicular to each other in the overlapping area.\",\n \"5.A device according to claim 1, wherein at least four flow channels (14, 16, 18, 20) extend from the first junction point (10), of which two join into the outer elongated first outlet openings (26, 28) that are arranged parallel to each other and the remaining join into additional outlet openings (32, 34) that run perpendicular in them, of which one, respectively, runs along the transverse peripheral region of the outer first outlet openings (26, 28).\",\n \"6.A device according to claim 1, wherein from the first junction point (10) a flow channel (18) extends; which joins both into one of the outer first outlet openings (28) and in additional outlet openings (40, 42, 44, 46) that run on the outside along its longitudinal edge.\"\n ],\n [\n \"The invention relates to a device for the production of an extruded profile, by extrusion of differing materials, particularly for the production of an extruded profile with at least three parallel and spaced bars, comprising at least two extruders that release the differing materials as well as flow channels leading to a tool that comprises outlet openings releasing the materials such as slots.\",\n \"In order to be able to produce semi-finished products or profile consisting of plastic continuously, extrusion processes are used.\",\n \"Here thermoplastic resins in the form of e.g.\",\n \"powder or granules are placed in an extruder, mixed with stabilizers, slip additives and possibly coloring and filling agents and are then conveyed to a profile-determining extrusion tool by means of a screw.\",\n \"During this process the materials are compressed, degassed and homogenized while simultaneously being mixed and kneaded.\",\n \"This creates a homogeneous, flowing mixture, the so-called molding compound, which is heated to or kept at a temperature that is characterized in the lower range by plastification and in the upper range by decomposition of the material.\",\n \"After traveling through an extrusion die specifying the geometry, the material runs through a calibration unit, which consists of a dry-wet calibration combination.\",\n \"Subsequent to that, the material is guided through a full bath vacuum or spray vacuum tank with integrated calibrating cover.\",\n \"A corresponding device is revealed in DE 197 44 515.Here the flow channels are united directly in front of a die front plate of the tool in order to produce an extruded profile.\",\n \"From EP 0 299 736 A1 we know of a co-extrusion tool in which also several flow channels run, which are united within the tool in order to produce a multi-layer extruded profile, which exhibits a laminate design.\",\n \"It is the object of the present invention to further develop a device of the afore-mentioned kind such that extruded profiles can be extruded, which can consist of differing material in order to be able to process in particular inexpensive, such as recycled, materials.\",\n \"Hereby it shall be ensured that the material that is supposed to be extruded flows to the necessary extent to relatively wide slots that are arranged parallel to one another.\",\n \"Pursuant to the invention the problem is essentially resolved in that the tool comprises at least three elongated, i.e.\",\n \"one inner and two outer, outlet slots, wherein the outer outlet slots each are connected to a first flow channel, for the supply of a first material, extending from a first junction point, and the inner outlet opening is connected to two second flow channels, for the supply of a second material, between which one of the first flow channels runs and which extend from one joint second junction point.\",\n \"Hereby the inner outlet slot and the outer outlet slots run parallel to each other.\",\n \"The junction points as such are connected to an extruder, respectively.\",\n \"To be able to produce even complex extruded profiles, one further development of the invention provides that further flow channels extend from the first junction point, which lead to additional preferably slot-shaped outlet openings running in the transverse peripheral region of at least one of the outer elongated outlet openings.\",\n \"In particular, at least one additional flow channel extends from the first junction point, wherein said channel ends up at another outlet opening that runs in a transverse peripheral region of one of the elongated outlet openings, in particular in a transverse peripheral region of the inner outlet opening.\",\n \"In a preferred embodiment, at least four flow channels extend from the first junction point, wherein two of said four channels lead into the outer elongated outlet openings and two lead perpendicular to them, of which one, respectively, extends along the transverse peripheral regions of the outer elongated outlet openings.\",\n \"The outlet openings as such join at a distance from each other into an outlet die of the tool so that the desired geometry of the profile can be achieved.\",\n \"Hereby inner rods, in particular the one extruded with the inner elongated outlet opening, can consist of less expensive resin such as recycled material.\",\n \"Further details, benefits and features of the invention are revealed not only in the claims, the characteristics disclosed therein—either alone and/or in combination—but also in the following description of an exemplary embodiment shown in the drawing.\",\n \"To produce profiles by means of extrusion, with said profiles consisting of sections comprising differing materials and relatively elongated bars that run parallel to each other in the range of for example 100 mm, pursuant to the invention a device with a tool is suggested, where the flow channels have the necessary diameter to guarantee material supply to outlet openings such as slots or sheet dies.\",\n \"The only FIGURE shows in purely principle form in a diagrammatic view flow channels that run in the tool of an extrusion device, whereby the channels extend from one of a total of two junction points 10, 12, which each are connected to an extruder, via which the flow channels are supplied with the flowing molding compound.\",\n \"In the example, four flow channels 14, 16, 18 and 20 extend from the first junction point 10 and two flow channels 22, 24 from the second junction point 12.As the basic diagram shows, the tool comprises three elongated outlet slots 26, 28, 30, which run parallel to each other, wherein the outer outlet slots 26, 28, which transition into corresponding outlet dies of the extrusion tool, are connected to the flow channels 16, 18 leading to the first junction point 10 and the inner outlet slot 30 is connected to the second junction point 12 via the flow channels 22, 24.In order to supply the outlet slot 30 to the necessary extent with flowing homogenized and compressed material, the outlet slot 30 is supplied via the two flow channels 22, 24 by the junction point 12, wherein the flow channels 22, 24 surround in some sections the flow channel 18 that leads to the upper outlet slot 30.In other words, the flow channels 22, 24 extend perpendicular or substantially perpendicular in the overlapping area to the flow channel 18.Due to the fact that the inner outlet slot 30 is connected to two flow channels 22, 24, sufficient molding compound can be supplied regardless of the small diameter of the flow channels 22, 24.In other words, due to the arrangement of the flow channels pursuant to the invention, the outlet slots 26, 28, 30 can be sufficiently supplied with the compressed flowing material.\",\n \"Furthermore, apart from the flow channels 16, 18 that lead to the outer elongated slotted dies 26, 28, two additional flow channels 14, 20 extend from the first junction point 10, which join into slots 32, 34 that run perpendicular to the outlet slots 26, 28, 30 and along their transverse edges.\",\n \"Moreover additional outlet openings 36, 38 or 40, 42, 44, 46 with a square or rectangular cross-section branch off the flow channels 14, 20 as well as off the flow channel 18 in order to be able to create the desired extruded profile.\",\n \"Hereby outlet openings 40, 42, 44, 46 are arranged along the outer longitudinal side of the upper outlet slot 28, i.e.\",\n \"on the side away from the inner slot 30.Due to the fact that the flow channels that extend from the second junction point 12 in some areas surround a flow channel 18 that extends from the first junction point 12, sufficient compressed flowing material can flow to the outlet slots 30 with optimal space utilization, whereby the junction points 10, 12 are connected to extruders by means of which the differing materials are plasticized.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10474197"}}},{"rowIdx":789,"cells":{"text":{"kind":"list like","value":[["Trolley wheel assembly","A trolley includes a front wheel assemblie (12) and a rear wheel assembly both assemblies mounted on a trolley frame.","The front wheel assembly (12) includes a fixed wheel assembly (24) to aid control of the trolley.","In one form the front wheel assembly (12) includes a pair of swivel castors (18) and a fixed wheel assembly (24) disposed between the castors (18).","The front wheel assembly (12) is designed so that when the castors (18) swivel from their orientation for forward travel, the fixed wheel assembly (24) is caused to be lifted off the ground.","The trolley also includes a camming device which is operative to lift the front wheel assembly (12) off the ground when the trolley is moved into a nested configuration with a like trolley."],["1.A trolley comprising a trolley frame, and front and rear wheel assemblies, one of the wheel assemblies including a pair of casters, each caster including a yoke mounted to the trolley frame in a manner that allows the caster to swivel about a first axis, and a caster wheel mounted to the yoke so as to be rotatable about a second axis which is transverse to said first axis, each caster being operative to adopt a first orientation for forward travel of said trolley, the one wheel assembly being configured so as to lift the trolley frame when the casters swivel about their first axes from the first orientation, wherein the one wheel assembly further comprises a fixed wheel assembly mounted to the trolley frame and including at least one wheel which has an axis of rotation which is orientated so that said fixed wheel assembly assists in maintaining directional control of said trolley in its forward travel, the wheel of the fixed wheel assembly being positioned to engage the ground when the castors are in the first orientation and to be off the ground when the trolley frame is lifted by swivelling of the casters about their first axes.","2.A trolley according to claim 1, wherein the one wheel assembly is configured to lift the trolley frame on swivelling of the casters from the first orientation by having the centre of the caster wheels radially displaced from their first axes and having the first axes inclined to the vertical and extending rearwardly of the trolley.","3.A trolley according to claim 1, wherein the fixed wheel assembly is disposed between the casters of the one wheel assembly.","4.A trolley according to claim 1, the one wheel assembly is the front wheel assembly.","5.A trolley according to claim 4, wherein the wheel of the fixed wheel assembly is aligned with the casters of the front wheel assembly.","6.A trolley according to claim 4 wherein the fixed wheel assembly is displaced either forward or rearward of the casters of the front wheel assembly.","7.A trolley according to claim 1, the other wheel assembly includes a pair of casters, each of the other wheel assembly casters including a yoke mounted to the trolley frame in a manner that allows the caster to swivel about a first axis, and a caster wheel mounted to the yoke so as to be rotatable about a second axis which is transverse to said first axis, each caster being operative to adopt a first orientation for forward travel of said trolley.","8.A trolley according to claim 1, wherein the fixed wheel assembly is the form of a fixed caster including a yoke mounted to the trolley frame, and a caster wheel rotatably mounted to the yoke.","9.A trolley comprising a trolley frame, and front and wheel assemblies, one of said wheel assemblies including a pair of casters each including a yoke mounted to the trolley frame which is operative to allow the casters to swivel about a first axis, and a caster wheel mounted to the yoke so as to be rotatable about a second axis which is transverse to said first axis, and an elongate member which extends through the respective yokes of said casters so as to maintain the casters in a first orientation for forward travel of said trolley and thereby prevent them swivelling about their respective first axes.","10.A trolley according to claim 9, wherein the elongate member forms the axle of the respective casters and thereby supports the caster wheels for rotational movement.","11.A trolley according to claim 9, wherein the trolley further comprises a fixed wheel assembly having at least one wheel and the elongate member forms the axle of the fixed wheel assembly and thereby supports the at least one wheel for rotational movement.","12.A trolley according to claim 11, wherein the caster wheels are smaller than the at least one wheel of the fixed wheel assembly.","13.A trolley according to claim 11, wherein the trolley is designed to be nested with another said trolley, wherein the front portion of one trolley is in overlapping relation with the rear portion of the other trolley, and wherein the trolley further includes lifting means which is operative to cause the wheels of the front wheel assembly of the one trolley to be lifted off the ground on nesting of the two trolleys.","14.A trolley according to claim 13, wherein the lifting means is in the form of complementary surfaces arranged to interengage during nesting of the trolleys.","15.A trolley according to claim 14, further comprising a camming device which includes a camming surface, the camming device being secured to the trolley so that the camming surface forms one of the complementary surfaces and engages a surface of the other trolley on nesting of the trolleys.","16.A trolley according to claim 15, wherein the trolley includes a basket having a rear gate, and wherein on nesting, the basket of the one trolley locates in the basket of the other trolley through the rear gate, and wherein the camming surface is positioned so as to engage a surface of the basket to cause lifting of the one trolley.","17.A trolley according to claim 16, wherein the camming device is mounted on a base surface of the basket and projects downwardly therefrom.","18.A trolley according to claim 16, the camming device is mounted to a base of the basket and projects upwardly therefrom.","19.A trolley according to claim 15, wherein the camming device comprises an elongate member having opposite ends and being profiled to form the camming surface intermediate the ends, the member further comprising engagement elements located at each end which are operative to be fixed to the basket of said trolley.","20.A trolley according to claim 19, wherein the basket of the trolley is formed from wire mesh and the engagement elements are in the form of eyelets which are operative to engage respective wire bars of the trolley basket.","21.A trolley comprising a trolley frame, and front and rear wheel assemblies mounted on said trolley frame, the trolley being designed to be nested with another trolley where a front portion of one trolley is in overlapping relation with a rear portion of the other trolley, and wherein the trolley further comprises lifting means operative to cause the wheels of the front wheel assembly of the one trolley to be lifted off the ground during nesting of the one trolley with the other trolley.","22.A camming device for a trolley, the camming device comprising an elongate member having opposite ends and being profiled to form a camming surface intermediate the ends, the member further comprising engagement elements located at each end that are operative to be fixed to the trolley.","23.A camming device according to claim 22, engagement elements are in the form of eyelets.","24.A camming device according to claim 22, wherein the camming device is made from sheet steel.","25.A method of retrofitting a trolley having front and rear wheel assemblies to incorporate a camming surface to improve nesting of the trolley with another trolley, the method comprising the steps of: providing a camming device comprising an elongate member having opposite ends and being profiled to form a camming surface intermediate the ends, and securing the camming device to the trolley in a position so that when the trolley is moved into a nested configuration with the other trolley, the camming surface is operative to engage the other trolley in the nested configuration so as to cause the rear trolley in the nested configuration to be lifted so that the front wheels of that trolley disengage the ground.","26.A method of securing a pair of casters mounted on a trolley body against swivelling, each caster having a wheel mounted on an axle passing through a yoke which is in turn rotatably mounted on said trolley body, said method including the steps of: providing an aperture in the yoke of each of said casters; positioning each caster wheel such that the rotation axes of the wheels are substantially coaxial; passing a rigid elongate member through each said aperture so as to maintain said rotational axis in a substantially coaxial relationship; and fixedly securing said elongate members to said yoke.","27.A pair of casters when secured against swivelling by a method according to claim 26.28.","(canceled) 29.","(canceled) 30.","(canceled) 31.","(canceled) 32.","(canceled) 33.","(canceled) 34.","(canceled)"],["<SOH> BACKGROUND OF THE INVENTION <EOH>Shopping trolleys are typically configured with four fully swivelling casters, one caster being provided at each corner of the base of the trolley frame to form front and rear wheel assemblies.","The use of four fully swivelling casters enables the shopping trolley to be manoeuvred forwards or sideways, however can render the trolley difficult to control, with steering a trolley around a corner or maintaining it in a straight path or on a sloped surface being particularly difficult.","With the casters being free to swivel and therefore not providing any sideways resistance, altering the path of the trolley requires the operator to impart a moment directly on the trolley to rotate it as there are no fixed wheels about which the trolley can easily rotate.","The difficulty in manoeuvring a trolley is increased when loaded with shopping, with the increased weight resulting in an increased moment being required to alter the path of the trolley.","The applicant has found that a shopping trolley having the front pair of wheels fixed from swivelling, with the wheel axles in a co-axial relationship, and the rear wheels being able to swivel, not unlike a motor vehicle, is easier to manoeuvre.","A trolley with such a configuration is able to pivot about the fixed set of wheels to alter its direction.","The operator need only control this pivoting about the fixed wheels by applying a small sideways force to the trolley handle to direct the trolley along the required path.","Whilst this arrangement is effective, the manoeuvrability of a trolley may be unduly limited, particularly when turning the trolley in tight situations."],["<SOH> SUMMARY OF THE INVENTION <EOH>Accordingly, an aim of the present invention is to ameliorate this problem whilst still utilising the advantages to trolleys of including a fixed wheel assembly.","In its broadest terms, the invention is directed to a trolley including a trolley frame, and front and rear wheel assemblies, one of said wheel assemblies including a pair of casters, each including a yoke mounted to the trolley frame in a manner that allows the caster to swivel about a first axis, and a caster wheel mounted to the yoke so as to be rotatable about a second axis which is transverse to said first axis, each caster being operative to adopt a first orientation for forward travel of said trolley, and a fixed wheel assembly located between said casters, said fixed wheel assembly including at least one wheel which has an axis of rotation which is orientated so that said fixed wheel assembly assists in maintaining directional control of said trolley in its forward travel.","In one form, the wheels of fixed wheel assembly and the casters in their first orientation are designed to engage the ground.","In this arrangement, the fixed wheel assembly provides the directional control for the trolley, whilst the casters which are able to swivel provide the lateral stability for the trolley to prevent tipping.","In another form, the wheel of fixed wheel assembly is designed to project below the level of the caster wheels when in their first orientation so that, in forward travel, the trolley is supported at said one wheel assembly by the fixed wheel assembly.","The casters which may be fixed or swivelling, act as outrigger wheels to prevent sideways tipping of the trolley.","In this latter arrangement, preferably the fixed wheel assembly includes a plurality of closely spaced wheels to provide more balanced support at the front of the trolley.","The arrangement of the invention has significant advantages.","By providing a central fixed wheel, enables the trolley to be manoeuvred more easily than is the case if the fixed wheels are widely spaced apart.","Further, it provides a system which can be easily and inexpensively retrofitted to existing trolleys.","It also maintains a useful purpose for the existing casters to enable the trolley to be stably supported.","In particular, the arrangement enables the trolley to meet some government requirements whereby a trolley must be supplied with at least four wheels.","In a particularly preferred form, when each caster adopts a second orientation which is angularly displaced from the first orientation about the first axis, the wheels of said casters are operative to extend below said wheel of said fixed wheel assembly so as to be operative to cause the wheel of the fixed wheel assembly to be off the ground.","Preferably the second orientation is angularly displaced from said first orientation by 90 degrees.","More preferably, the fixed wheel is operative to be lifted off the ground when either caster is displaced angularly from the first orientation by more than 20 degrees.","Preferably, the caster wheel in each caster is offset from its first axis so that, in use, the caster wheel swivels about an arc around its first axis.","In addition, preferably the caster is mounted to the trolley frame with the first axis inclined to the vertical so that in use, once swivelling of the casters about their respective first axes, the vertical displacement of the centre of the respective casters from a fixed point on the trolley varies.","In a particularly preferred form, each caster is mounted to the trolley so that this vertical displacement is at a minimum when that caster is in its first orientation.","This arrangement has particular advantage in that when either of the casters is caused to be angularly spaced about its first axis from the first orientation by a predetermined amount, that caster wheel is lower than the fixed wheel thereby lifting the fixed wheel off the ground.","The degree of angular displacement required to provide adequate clearance to fully lift the fixed wheel off the ground will depend on the specific configuration of the wheel assembly, but the applicant has found that displacement of 10-20 degrees from the first orientation is ideal.","Further, if the casters are operative to remain in contact with the ground at all times, they can be caused to swivel from the first orientation (and thereby lift the fixed wheel off the ground) simply by applying a sideways force to the trolley.","This arrangement has the advantage that the fixed wheel assembly assists in controlling forward movement of the trolley, but does not inhibit sideways movement of the trolley as the caster wheels, as they pivot, would be caused to lift this fixed wheel assembly off the ground.","In addition, the fixed wheel can inhibit the trolley inadvertently “running away” when it is stationery as it lessens the tendency to move sideways if parked on a sideways sloping surface.","In one arrangement, the novel wheel assembly may be retrofitted to an existing trolley assembly by securing a fixed wheel assembly to the trolley body.","This may be secured to the existing cross bar supporting the caster wheel assemblies or an additional cross bar may be installed.","Further, the casters may be arranged to have their axes inclined to the vertical in the desired orientation merely by tilting the trolley frame which can be simply achieved by placing larger casters on the rear wheel assembly.","Alternatively the caster swivel axis may be inclined by the addition of an appropriately tapered spacers mounted at the point of attachment of the caster to the trolley.","In another form, an elongate rigid member is used to interconnect the frames of the caster wheel assemblies.","In this latter arrangement, preferably the elongate member is used as the axle for the fixed wheel assembly.","Appropriate spaces, or fixtures are provided on the axle to properly locate those fixed wheels in place.","In a particularly preferred form of this latter arrangement, the elongate member extends through the frame axles of the respective casters.","Further, the caster wheels from the pair of assemblies are removed and are used as part of the fixed wheel assembly.","In their place, smaller outrigger wheels are used.","The advantage of this arrangement is that the caster wheels are designed to be constantly in contact with the ground.","As such, they are ideally suited to be used as the fixed wheel assembly.","Further as the outrigger wheels are only used to prevent tipping of the trolley, an inferior wheel may be used as compared to the caster wheels, thereby further reducing the cost of retrofitting.","In yet a further aspect, the invention is directed to a method of retrofitting a trolley using any one of the techniques described above.","In a particularly preferred form, the front wheel assembly includes the fixed wheel assembly.","In this arrangement preferably the rear wheel assembly includes pivoting casters, although other arrangements may be used.","Whilst the design of the shopping trolley with a fixed wheel as part of the front wheel assembly aids control of an individual trolley, it may impede the manoeuvrability of the trolleys when in a nested configuration as a train as it significantly restricts the ability to be able to change the direction of the train of trolleys.","Accordingly to a further aspect, the present invention provides a trolley including a trolley frame, and front and rear wheel assemblies mounted on said trolley frame, the trolley being designed to be nested with a like trolley where a front portion of one said trolley is in overlapping relation with a rear portion of another said trolley, and wherein the trolley further includes lifting means which is operative to cause the wheels of the front wheel assembly of the one trolley to be lifted off the ground during nesting of the one trolley with said other trolley.","A trolley according to the above form has significant advantage.","The trolley may exhibit improved manoeuvrability by having a fixed wheel assembly at the front wheel assembly according to any form described above.","Further, the manoeuvrability of a train of nested trolleys in accordance with the invention is also significantly easier to manoeuvre by virtue of the lifting means which disengages each of the front wheels assemblies of the nested trolleys off the ground.","With this arrangement, only the front wheel assembly of the front trolley in the train is in engagement with the ground.","Therefore the train includes one fixed front wheel assembly with the remaining wheels being part of the individual trolleys rear wheels which are preferably casters.","This provides an arrangement which is easier to manoeuvre than traditional trains of nested shopping trolleys which includes only swivelling casters.","Further, by virtue of the fact that the front wheel assemblies of the nested trolleys is disengaged from the ground, the number of wheels actually in contact with the ground in any train is significantly reduced which further improves the manoeuvrability of the train of nested trolleys.","In a preferred form, the lifting means is in the form of a camming surface arranged to engage a complementary surface on the like trolley during nesting.","Through that engagement, the wheels on the front wheel assembly of the one trolley is caused to move off the ground.","This camming arrangement may be located on any suitable part of the trolley such as on the basket, or trolley frame.","In an alternative form, the front wheel assembly may be operative to move out of engagement with the ground using a different mechanism as the lifting means.","For example, the wheel assemblies may be pivotable from an extended to a retracted configuration, and wherein on nesting of the one trolley with the like trolley, the front wheel assemblies may be caused to move from their extended position to their retracted position.","In yet a further aspect, the present invention relates to the retrofitting of existing shopping trolleys so as to provide the enhanced manoeuvrability afforded by the shopping trolleys of the earlier aspects of the invention.","In particular, in one aspect the invention is directed to retrofitting existing shopping trolleys so as to incorporate the lifting means.","This aspect aims to use techniques which are inexpensive which thereby will not be prohibitively expensive to retrofit hundreds of shopping trolleys in a typical supermarket.","In accordance with this further aspect, the invention relates to a camming device for a trolley, the camming device including a body incorporating a camming surface, and attachment means operative to fix the camming device to said shopping trolley, the trolley including a basket incorporating a rear gate and the trolley being designed so as to be able to adopt a nested configuration with a like trolley, where the basket of one trolley locate through the rear gate of another trolley so that when nested, a substantial portion of the basket of the one trolley is disposed within the basket of the like trolley, and wherein in use, when the camming device is fixed to the trolley, the camming surface is operative to engage the other trolley in the nested configuration so as to lift the one trolley as it is moved into the nested configuration so as to cause the front wheel assembly to disengage the ground.","In a preferred form, the camming device includes an elongate member having opposite ends, the member being shaped so as to form a camming surface intermediate the ends, and wherein the attachment means comprises engagement elements located at each end which are operative to be fixed to the trolley.","In a preferred form, the camming device is operative to be secured to the basket of the trolley and the engagement means is preferably in the form of eyelets which are operative to engage respective wire bars of the trolley basket.","The advantage of this arrangement is that the camming device is inexpensive, and the method of attachment can be easily performed by providing the eyelets in partially open form on respective wire bars of the basket and merely clinching them to securely affix them in place.","In yet a further aspect, the present invention relates to a method of retrofitting a trolley to incorporate a camming device which includes the steps of providing a camming device in any form described above, and securing the camming device to the trolley in a position so that when the trolley is moved into a nested configuration with a like trolley, the camming device on one of the trolleys is operative to engage the other of the trolley in the nested configuration so as to cause the one trolley to be lifted so that the front wheels disengage the ground in the nested configuration.","It is to be appreciated that the trolley including the lifting means in accordance with these aspects of the invention is ideally suited to be used in conjunction with a trolley including the front wheel assembly with a fixed wheel assembly in accordance with the earlier aspects of the invention.","Alternatively, it may be used with trolleys having other arrangements of front wheel assemblies, whether they include a fixed wheel or not.","In yet a further aspect, the invention relates to methods and systems to modify an existing pair of caster wheel assemblies so as to provide a fixed axle wheel arrangement using simple and inexpensive techniques.","Accordingly, in this aspect, there is disclosed herein apparatus adapted to secure a pair of casters mounted on a trolley body against swivelling, each said caster having a wheel mounted on an axle passing through a yoke which is in turn rotatably mounted on said trolley body, said apparatus including: a rigid elongate member adapted to pass through an aperture provided in each said yoke such that the rotation axes of said wheels are substantially coaxial, and means for fixedly securing said elongate member to said yokes.","Typically said elongate member has a circular cross-section toward each end therefor and is adapted to replace the axle of each of said casters.","Typically said securing means includes a pair of nuts each respectively adapted to thread onto a threaded portion provided at each respective end of said elongate member so as to abut the respective said yoke.","Typically said securing means includes means for abutting each said yoke on an opposing side thereof to the associated elongate member end.","Preferably said abutting means includes a pair of circlips each adapted to be received in a groove provided in said elongate member, each said groove being positioned such that, in use, each said circlip abuts a said frame.","Preferably said abutting means includes a pair of shoulders each defined by a change in cross-section of said elongate member.","Alternatively the apparatus further includes a sleeve locatable over the elongate member and wherein opposite ends of the sleeve provides said abutting means.","There is further disclosed herein a method of securing a pair of casters mounted on a trolley body against swivelling, each said caster having a wheel mounted on an axle passing through a yoke which is in turn rotatably mounted on said trolley body, said method including the steps of: providing an aperture in the yoke of each of said caster wheel assemblies; positioning each said caster such that the rotation axes of said wheels are substantially co-axial; passing a rigid elongate member through each said aperture so as to maintain said rotation axes in a substantially co-axial relationship, and fixedly securing said elongate member to said yokes.","Typically said elongate member is circular in cross-section towards each end thereof and said step of providing an aperture in each yoke is achieved by removing the axle of each said caster, said elongate member replacing said axles so as to provide said axes of rotation of said wheels.","There is still further disclosed herein a method of modifying the handling characteristics of a shopping trolley having four caster wheel assemblies mounted on the body thereof, said method including the step of securing either the front or rear pair of caster wheels against swivelling according to the above disclosed method.","There is still further disclosed herein a trolley modified by the above method.","Accordingly, the present invention provides a novel trolley and associated methods of modification of existing trolleys, which has the advantage of the fixed front wheels, but have improved manoeuvrability, and which may be inexpensively retrofitted to be incorporated into existing trolleys."],["TECHNICAL FIELD The present invention relates to a trolley and in particular to a trolley with a modified wheel assembly to aid in control of the trolley.","The invention has been designed especially, but not exclusively, for use in shopping trolleys and the invention is herein described in that context.","It is to be appreciated however that the invention is not limited to that use.","BACKGROUND OF THE INVENTION Shopping trolleys are typically configured with four fully swivelling casters, one caster being provided at each corner of the base of the trolley frame to form front and rear wheel assemblies.","The use of four fully swivelling casters enables the shopping trolley to be manoeuvred forwards or sideways, however can render the trolley difficult to control, with steering a trolley around a corner or maintaining it in a straight path or on a sloped surface being particularly difficult.","With the casters being free to swivel and therefore not providing any sideways resistance, altering the path of the trolley requires the operator to impart a moment directly on the trolley to rotate it as there are no fixed wheels about which the trolley can easily rotate.","The difficulty in manoeuvring a trolley is increased when loaded with shopping, with the increased weight resulting in an increased moment being required to alter the path of the trolley.","The applicant has found that a shopping trolley having the front pair of wheels fixed from swivelling, with the wheel axles in a co-axial relationship, and the rear wheels being able to swivel, not unlike a motor vehicle, is easier to manoeuvre.","A trolley with such a configuration is able to pivot about the fixed set of wheels to alter its direction.","The operator need only control this pivoting about the fixed wheels by applying a small sideways force to the trolley handle to direct the trolley along the required path.","Whilst this arrangement is effective, the manoeuvrability of a trolley may be unduly limited, particularly when turning the trolley in tight situations.","SUMMARY OF THE INVENTION Accordingly, an aim of the present invention is to ameliorate this problem whilst still utilising the advantages to trolleys of including a fixed wheel assembly.","In its broadest terms, the invention is directed to a trolley including a trolley frame, and front and rear wheel assemblies, one of said wheel assemblies including a pair of casters, each including a yoke mounted to the trolley frame in a manner that allows the caster to swivel about a first axis, and a caster wheel mounted to the yoke so as to be rotatable about a second axis which is transverse to said first axis, each caster being operative to adopt a first orientation for forward travel of said trolley, and a fixed wheel assembly located between said casters, said fixed wheel assembly including at least one wheel which has an axis of rotation which is orientated so that said fixed wheel assembly assists in maintaining directional control of said trolley in its forward travel.","In one form, the wheels of fixed wheel assembly and the casters in their first orientation are designed to engage the ground.","In this arrangement, the fixed wheel assembly provides the directional control for the trolley, whilst the casters which are able to swivel provide the lateral stability for the trolley to prevent tipping.","In another form, the wheel of fixed wheel assembly is designed to project below the level of the caster wheels when in their first orientation so that, in forward travel, the trolley is supported at said one wheel assembly by the fixed wheel assembly.","The casters which may be fixed or swivelling, act as outrigger wheels to prevent sideways tipping of the trolley.","In this latter arrangement, preferably the fixed wheel assembly includes a plurality of closely spaced wheels to provide more balanced support at the front of the trolley.","The arrangement of the invention has significant advantages.","By providing a central fixed wheel, enables the trolley to be manoeuvred more easily than is the case if the fixed wheels are widely spaced apart.","Further, it provides a system which can be easily and inexpensively retrofitted to existing trolleys.","It also maintains a useful purpose for the existing casters to enable the trolley to be stably supported.","In particular, the arrangement enables the trolley to meet some government requirements whereby a trolley must be supplied with at least four wheels.","In a particularly preferred form, when each caster adopts a second orientation which is angularly displaced from the first orientation about the first axis, the wheels of said casters are operative to extend below said wheel of said fixed wheel assembly so as to be operative to cause the wheel of the fixed wheel assembly to be off the ground.","Preferably the second orientation is angularly displaced from said first orientation by 90 degrees.","More preferably, the fixed wheel is operative to be lifted off the ground when either caster is displaced angularly from the first orientation by more than 20 degrees.","Preferably, the caster wheel in each caster is offset from its first axis so that, in use, the caster wheel swivels about an arc around its first axis.","In addition, preferably the caster is mounted to the trolley frame with the first axis inclined to the vertical so that in use, once swivelling of the casters about their respective first axes, the vertical displacement of the centre of the respective casters from a fixed point on the trolley varies.","In a particularly preferred form, each caster is mounted to the trolley so that this vertical displacement is at a minimum when that caster is in its first orientation.","This arrangement has particular advantage in that when either of the casters is caused to be angularly spaced about its first axis from the first orientation by a predetermined amount, that caster wheel is lower than the fixed wheel thereby lifting the fixed wheel off the ground.","The degree of angular displacement required to provide adequate clearance to fully lift the fixed wheel off the ground will depend on the specific configuration of the wheel assembly, but the applicant has found that displacement of 10-20 degrees from the first orientation is ideal.","Further, if the casters are operative to remain in contact with the ground at all times, they can be caused to swivel from the first orientation (and thereby lift the fixed wheel off the ground) simply by applying a sideways force to the trolley.","This arrangement has the advantage that the fixed wheel assembly assists in controlling forward movement of the trolley, but does not inhibit sideways movement of the trolley as the caster wheels, as they pivot, would be caused to lift this fixed wheel assembly off the ground.","In addition, the fixed wheel can inhibit the trolley inadvertently “running away” when it is stationery as it lessens the tendency to move sideways if parked on a sideways sloping surface.","In one arrangement, the novel wheel assembly may be retrofitted to an existing trolley assembly by securing a fixed wheel assembly to the trolley body.","This may be secured to the existing cross bar supporting the caster wheel assemblies or an additional cross bar may be installed.","Further, the casters may be arranged to have their axes inclined to the vertical in the desired orientation merely by tilting the trolley frame which can be simply achieved by placing larger casters on the rear wheel assembly.","Alternatively the caster swivel axis may be inclined by the addition of an appropriately tapered spacers mounted at the point of attachment of the caster to the trolley.","In another form, an elongate rigid member is used to interconnect the frames of the caster wheel assemblies.","In this latter arrangement, preferably the elongate member is used as the axle for the fixed wheel assembly.","Appropriate spaces, or fixtures are provided on the axle to properly locate those fixed wheels in place.","In a particularly preferred form of this latter arrangement, the elongate member extends through the frame axles of the respective casters.","Further, the caster wheels from the pair of assemblies are removed and are used as part of the fixed wheel assembly.","In their place, smaller outrigger wheels are used.","The advantage of this arrangement is that the caster wheels are designed to be constantly in contact with the ground.","As such, they are ideally suited to be used as the fixed wheel assembly.","Further as the outrigger wheels are only used to prevent tipping of the trolley, an inferior wheel may be used as compared to the caster wheels, thereby further reducing the cost of retrofitting.","In yet a further aspect, the invention is directed to a method of retrofitting a trolley using any one of the techniques described above.","In a particularly preferred form, the front wheel assembly includes the fixed wheel assembly.","In this arrangement preferably the rear wheel assembly includes pivoting casters, although other arrangements may be used.","Whilst the design of the shopping trolley with a fixed wheel as part of the front wheel assembly aids control of an individual trolley, it may impede the manoeuvrability of the trolleys when in a nested configuration as a train as it significantly restricts the ability to be able to change the direction of the train of trolleys.","Accordingly to a further aspect, the present invention provides a trolley including a trolley frame, and front and rear wheel assemblies mounted on said trolley frame, the trolley being designed to be nested with a like trolley where a front portion of one said trolley is in overlapping relation with a rear portion of another said trolley, and wherein the trolley further includes lifting means which is operative to cause the wheels of the front wheel assembly of the one trolley to be lifted off the ground during nesting of the one trolley with said other trolley.","A trolley according to the above form has significant advantage.","The trolley may exhibit improved manoeuvrability by having a fixed wheel assembly at the front wheel assembly according to any form described above.","Further, the manoeuvrability of a train of nested trolleys in accordance with the invention is also significantly easier to manoeuvre by virtue of the lifting means which disengages each of the front wheels assemblies of the nested trolleys off the ground.","With this arrangement, only the front wheel assembly of the front trolley in the train is in engagement with the ground.","Therefore the train includes one fixed front wheel assembly with the remaining wheels being part of the individual trolleys rear wheels which are preferably casters.","This provides an arrangement which is easier to manoeuvre than traditional trains of nested shopping trolleys which includes only swivelling casters.","Further, by virtue of the fact that the front wheel assemblies of the nested trolleys is disengaged from the ground, the number of wheels actually in contact with the ground in any train is significantly reduced which further improves the manoeuvrability of the train of nested trolleys.","In a preferred form, the lifting means is in the form of a camming surface arranged to engage a complementary surface on the like trolley during nesting.","Through that engagement, the wheels on the front wheel assembly of the one trolley is caused to move off the ground.","This camming arrangement may be located on any suitable part of the trolley such as on the basket, or trolley frame.","In an alternative form, the front wheel assembly may be operative to move out of engagement with the ground using a different mechanism as the lifting means.","For example, the wheel assemblies may be pivotable from an extended to a retracted configuration, and wherein on nesting of the one trolley with the like trolley, the front wheel assemblies may be caused to move from their extended position to their retracted position.","In yet a further aspect, the present invention relates to the retrofitting of existing shopping trolleys so as to provide the enhanced manoeuvrability afforded by the shopping trolleys of the earlier aspects of the invention.","In particular, in one aspect the invention is directed to retrofitting existing shopping trolleys so as to incorporate the lifting means.","This aspect aims to use techniques which are inexpensive which thereby will not be prohibitively expensive to retrofit hundreds of shopping trolleys in a typical supermarket.","In accordance with this further aspect, the invention relates to a camming device for a trolley, the camming device including a body incorporating a camming surface, and attachment means operative to fix the camming device to said shopping trolley, the trolley including a basket incorporating a rear gate and the trolley being designed so as to be able to adopt a nested configuration with a like trolley, where the basket of one trolley locate through the rear gate of another trolley so that when nested, a substantial portion of the basket of the one trolley is disposed within the basket of the like trolley, and wherein in use, when the camming device is fixed to the trolley, the camming surface is operative to engage the other trolley in the nested configuration so as to lift the one trolley as it is moved into the nested configuration so as to cause the front wheel assembly to disengage the ground.","In a preferred form, the camming device includes an elongate member having opposite ends, the member being shaped so as to form a camming surface intermediate the ends, and wherein the attachment means comprises engagement elements located at each end which are operative to be fixed to the trolley.","In a preferred form, the camming device is operative to be secured to the basket of the trolley and the engagement means is preferably in the form of eyelets which are operative to engage respective wire bars of the trolley basket.","The advantage of this arrangement is that the camming device is inexpensive, and the method of attachment can be easily performed by providing the eyelets in partially open form on respective wire bars of the basket and merely clinching them to securely affix them in place.","In yet a further aspect, the present invention relates to a method of retrofitting a trolley to incorporate a camming device which includes the steps of providing a camming device in any form described above, and securing the camming device to the trolley in a position so that when the trolley is moved into a nested configuration with a like trolley, the camming device on one of the trolleys is operative to engage the other of the trolley in the nested configuration so as to cause the one trolley to be lifted so that the front wheels disengage the ground in the nested configuration.","It is to be appreciated that the trolley including the lifting means in accordance with these aspects of the invention is ideally suited to be used in conjunction with a trolley including the front wheel assembly with a fixed wheel assembly in accordance with the earlier aspects of the invention.","Alternatively, it may be used with trolleys having other arrangements of front wheel assemblies, whether they include a fixed wheel or not.","In yet a further aspect, the invention relates to methods and systems to modify an existing pair of caster wheel assemblies so as to provide a fixed axle wheel arrangement using simple and inexpensive techniques.","Accordingly, in this aspect, there is disclosed herein apparatus adapted to secure a pair of casters mounted on a trolley body against swivelling, each said caster having a wheel mounted on an axle passing through a yoke which is in turn rotatably mounted on said trolley body, said apparatus including: a rigid elongate member adapted to pass through an aperture provided in each said yoke such that the rotation axes of said wheels are substantially coaxial, and means for fixedly securing said elongate member to said yokes.","Typically said elongate member has a circular cross-section toward each end therefor and is adapted to replace the axle of each of said casters.","Typically said securing means includes a pair of nuts each respectively adapted to thread onto a threaded portion provided at each respective end of said elongate member so as to abut the respective said yoke.","Typically said securing means includes means for abutting each said yoke on an opposing side thereof to the associated elongate member end.","Preferably said abutting means includes a pair of circlips each adapted to be received in a groove provided in said elongate member, each said groove being positioned such that, in use, each said circlip abuts a said frame.","Preferably said abutting means includes a pair of shoulders each defined by a change in cross-section of said elongate member.","Alternatively the apparatus further includes a sleeve locatable over the elongate member and wherein opposite ends of the sleeve provides said abutting means.","There is further disclosed herein a method of securing a pair of casters mounted on a trolley body against swivelling, each said caster having a wheel mounted on an axle passing through a yoke which is in turn rotatably mounted on said trolley body, said method including the steps of: providing an aperture in the yoke of each of said caster wheel assemblies; positioning each said caster such that the rotation axes of said wheels are substantially co-axial; passing a rigid elongate member through each said aperture so as to maintain said rotation axes in a substantially co-axial relationship, and fixedly securing said elongate member to said yokes.","Typically said elongate member is circular in cross-section towards each end thereof and said step of providing an aperture in each yoke is achieved by removing the axle of each said caster, said elongate member replacing said axles so as to provide said axes of rotation of said wheels.","There is still further disclosed herein a method of modifying the handling characteristics of a shopping trolley having four caster wheel assemblies mounted on the body thereof, said method including the step of securing either the front or rear pair of caster wheels against swivelling according to the above disclosed method.","There is still further disclosed herein a trolley modified by the above method.","Accordingly, the present invention provides a novel trolley and associated methods of modification of existing trolleys, which has the advantage of the fixed front wheels, but have improved manoeuvrability, and which may be inexpensively retrofitted to be incorporated into existing trolleys.","BRIEF DESCRIPTION OF THE DRAWINGS It is convenient to hereinafter describe embodiments of the present invention with reference to the accompanying drawings.","It is to be appreciated that the particularity of the drawings and the related description is to be understood as not superseding the generality of the preceding broad description of the invention.","In the drawings: FIG.","1 is a side view of a shopping trolley according to a first embodiment of the invention; FIG.","2 is a front elevation view of a front wheel assembly of the trolley of FIG.","1; FIGS.","3A and 3B are detailed side elevations of the front wheel assembly of FIG.","2; FIG.","4 is a front elevation view of a front wheel assembly for a trolley according to a second embodiment of the invention; FIG.","5 is a detailed view of the camming device in the trolley of FIG.","1; FIG.","6 is a side view of the trolley of FIG.","1 in a nested configuration; FIG.","7 is a front elevation view of a pair of casters mounted on a trolley body and modified with the apparatus of a further embodiment of the invention; FIG.","8 is a side elevation view of the casters of FIG.","7; FIG.","9 is a side elevation view of a pair of casters modified in an alternative manner; FIG.","10 is a front elevation view of an elongate rod of the apparatus of FIG.","7; and FIG.","11 is a front elevation view of an elongate rod of the apparatus of FIG.","9.DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS A shopping trolley 10 includes a frame 11 from which the front wheel assembly 12 and a rear wheel assembly 13 is mounted.","A wire basket 14 is supported on the frame and includes a handle 15 and rear gate 16.As is typical in shopping trolleys, the rear wheel assembly includes a pair of swivel casters 17 mounted at each back corner of the frame 11.As best illustrated in FIGS.","2, 3A and 3B, the front wheel assembly 12 include a pair of swivel casters which are mounted on respective front corners of the frame 11.Each caster 18 includes a yoke 19 which is arranged to rotate about a king pin 20 allowing the casters 18 to rotate about respective first axes 21.Caster wheels 22 are mounted on the respective yokes 19 and are rotatable about respective second axes 23 which are transverse to their respective first axes 21.To provide control to the trolley, the front wheel assembly 12 include a centre fixed wheel assembly 24 which is mounted to the trolley frame 11.In the illustrated form, the fixed wheel assembly 24 is displaced rearwardly of the front wheel assembly and is secured by a bracket 25 which is secured to the frame 11.In the illustrated form, the fixed wheel assembly 24 is secured to the bracket 25 by locating a threaded bolt 26 through an aperture in the bracket 25 which is secured in place by a nut 27.It is to be appreciated that other forms of fixing could be employed as will be appreciated by those skilled in the art.","Further, the fixed wheel assembly 24 may be located in line with the casters 18 if there is adequate clearance to allow those casters to swivel, or disposed slightly forward.","The fixed wheel assembly 24 is in the form of a non swivelling caster and includes a yoke 28 and a wheel 29 which is mounted to the yoke 28 and rotatable about an axis of rotation 30.FIGS.","3A and 3B are a side elevation of the front wheel assembly of FIG.","1.As can be seen, in this arrangement, the fixed wheel assembly 24 is secured rearward of the line of the casters 18.In addition, the first axes 21 of the casters 18 are each inclined to the vertical (V).","This is achieved in the illustrated embodiment by having the trolley frame 11 slightly inclined forwardly through having larger casters on the rear wheel assembly than the front wheel assembly.","Further, as illustrated in FIG.","3A, the centre line (P) of each wheel 22 is displaced from its first axis 21.As such, the caster wheels 22 are designed to move in an arc around their first axes 21.Because each first axis 21 is inclined to the vertical, the plane in which the caster wheels will move on swivelling is inclined to the horizontal plane.","In the illustrated embodiment, the first axes 21 of the casters 18 are inclined in the forward direction of the trolley.","As a result of this orientation, when the casters 18 are in a trailing position for forward movement of the trolley as illustrated in FIG.","3A, the caster wheels 22 are at the highest point of their arc.","Further, the fixed wheel 29 of the trolley is designed so that it is aligned with the height of the caster wheels at this point so that both the caster wheels 18 and the fixed wheel 29 are engaging with the surface 200 on forward movement of the trolley.","When the caster wheels 100 swivel about their first axes 21 away from the trailing position as shown in FIG.","3A, the distance between the caster wheel centres and the trolley frame 11 increases, which causes the trolley frame 11 to lift.","This, in turn, causes the fixed wheel 29 to move off the ground as illustrated in FIG.","3B.","This arrangement has significant practical benefit as it enables the fixed wheel 30 to assist in fully maintaining control of the trolley in the forward direction but still enables easy sideways movement of the trolley if required.","Accordingly, front wheel assembly 12 has the advantages of a centralised fixed wheel for pivoting and manoeuvrability of the trolley particularly for forward movement, but which still can provide adequate stability to the trolley to prevent tipping.","Further, the design of the front wheel assembly allows unimpeded sideways movement by creating a system where the fixed wheel can disengage the ground on certain orientations of the front casters.","FIG.","4 illustrates a modified version of the front wheel assembly 12.Again the main principal of the wheel assembly 12 is to provide a centralised fixed wheel between the front casters 18 to facilitate manoeuvrability of the trolley.","In this arrangement however, the casters 18 are modified as part of the installation of the fixed wheel assembly 31.In particular in the arrangement of FIG.","4, the caster wheels 22 are removed from their respective yoke 19 and those yokes are secured against swivelling by a rigid elongate member 32.Outrigger wheels 34 are included in the modified assembly 12 and the caster wheels 22 form part of the fixed centre wheel assembly 31.In constructing the front wheel assembly 12, an aperture is provided in each of the yokes 19 through which the elongate member 32 may pass when the casters 18 are positioned with the second axis 23 substantially coaxial.","Individual axle in each caster 18 is removed so that the elongate member is passed through the axle apertures 35 provided in the respective yokes 19.In the illustrated form, the elongate member 32 includes a threaded portion 36 at each opposite end for receiving a nut 37 to secure the elongate member 32 in place.","In conjunction with installing the elongate member 32 to actually fix the yokes 19, the caster wheels 22 are removed from the yokes 19 and are mounted on the member 32 intermediate the casters 18.Spaces 38 which in the illustrated form are sleeves of metal or plastic, are disposed on the elongate member to locate the caster wheels 22 in their central position.","In this way, the elongate member which has a circular cross section provides the axis of rotation for the wheels 22 which are now centrally fixed.","It is to be appreciated that other suitable spaces, such as circlips or the like may be used.","In addition, outrigger wheels 34 are installed in the yokes 19 to provide additional stability to the trolley.","These outrigger wheels are smaller than the casters 22 and are therefore only designed to come in contact with the ground should the trolley 10 tip.","The trolley 10 in any form described above is designed to nest with a like trolley 10l as best shown in FIG.","6.Specifically the trolley basket 14 with the pivoting rear gate 16, tapers towards its front.","With this arrangement, the trolley 10 is able to nest with the like trolley 101 with the basket 14 locating through the rear gate 16l of the like trolley 10l so that when nested, the basket 14 of the trolley is disposed within the basket 14l of the like trolley.","The trolley 10 includes a camming device 40 which is mounted to the basket 16.The device 40 includes an elongate metal member 41 which is arcuately shaped so as to form a cam surface 42 over its mid region.","Each end (44,46) of the member 41 includes rolled over edges to form eyelets 45, 46.Wire bars 47 of the basket 16 are operative to locate within the eyelets 45, 46 so as to secure the camming device to the basket.","To retrofit the camming device to the shopping trolley 10, the member 41 is provided with the eyelets partially opened so as to enable respective bars 47 of the basket 14 to be received in the eyelets whereafter they are secured in place by clinching the respective eyelets to close them.","In fitting the camming device 40 in place, the shopping trolley 10 may be provided with a single device, or multiple devices may be provided.","Further, in the illustrated form, the camming devices 40 depends downwardly from the basket 14.The advantage of this arrangement is that it places the camming device out of the way and does not reduce the volume of the material which can be provided within the trolley basket 14.Further, the formation of eyelets 45,46 at the end of the member provides a convenient safety edge to the camming device 20 which is unlikely to provide a pinch point or sharp edge which could otherwise catch on material placed in the basket 14.The inclusion of the camming device 40 is designed to cause a shopping trolley 10l when moved into a nested configuration to lift to a sufficient degree to cause the front wheel assembly 12l of nested trolley 10l to disengage the ground.","This is achieved by the camming surface 22 moving into engagement with the bottom surface 481 of the trolley basket 141.By virtue of this engagement of surfaces, the trolley 10l is caused to lift disengaging the front wheel assemblies 12l from the ground.","It is to be appreciated that the camming device 40 could be disposed to extend upwardly from the base of the trolley basket 14 and achieve the same result when a trolley is moved into a nesting configuration.","When in this nested configuration, the lifting of the front wheel assemblies 12l ensures that these assemblies do not impede the manoeuvrability of a train of the trolleys in a nested configuration.","FIGS.","7 to 11 disclose yet a further form of modifying the casters of the front wheel assembly 12.This embodiment has many similarities to the embodiment shown with reference to FIG.","4 with the exception that the modified trolley does not include a central fixed wheel assembly.","As best shown in FIG.","7, the wheels 22 of the casters 18 are secured against swivelling by a rigid elongate member 50.An aperture is provided in each of the yokes 19 through which the elongate member 50 may pass when the casters 18 are positioned with the rotation axes of the wheels 22 substantially co-axial.","In one embodiment, the axles 51 of each wheel are removed so that the elongate member is passed through the axle apertures 52 provided in the yokes 19.The elongate member 50, whose ends have a circular cross-section, thus replaces the axles so as to provide the axes of rotation of the wheels 22, ensuring that the axes of rotation are co-axial.","The elongate member 50 is then fixedly secured to the yokes 19.The elongate member 50 of this embodiment, which is in the form of a circular rod, is best depicted in FIG.","10.A threaded portion 53 is provided at each opposing end of the elongate rod 50 for receiving a nut 54.A nut 54 is threaded onto each threaded portion 53 once the elongate rod has passed through the apertures 52.Each nut 54 is tightened onto the threaded portion 53 until it abuts the associated yoke 19 adjacent the elongate frame end.","A washer 55 is typically provided between the nut 54 and yoke 19.Fastening the nuts 54 onto each end of the elongate rod 50 thus fixedly secures the elongate rod 50, with each end secured to its respective yoke 19.The rotation axes of the wheels 22 are thus fixed in the substantially co-axial relationship.","To further secure the elongate rod 50, means may be provided to abut each yoke 19 on an opposing side thereof to the associated elongate member end.","In the first embodiment this means is provided by circlips 56.The circlips 56 each abut an inner fork 62 of yoke 19, whilst a nut 54 abuts an outer fork 63 on the opposing side of the yoke 19.With each yoke 19 being abutted on opposing sides, the securing thereof is more robust.","Each circlip 56 is received in a circumferential groove 57 provided in the elongate member 50 at a position adjacent to the inner fork 62 of each yoke 19 when the elongate member 50 is in position.","An alternative elongate member 58 is depicted in FIG.","11.Like elongate member 50 it is provided with threaded portions 59 for receiving nuts 57.However, in place of the grooves 57 for receiving circlips, shoulders 60 are provided at positions corresponding to adjacent the inner fork 62 of each yoke 19.The shoulders 60 are formed by a change in cross-section of the elongate member 58 from a larger cross-section in a central region to smaller cross-sections at outer regions of the elongate member.","These shoulders 60 thus abut the inner forks 62 when the elongate member 58 is in position, helping to further secure the yokes 19 against swivelling.","Rather than providing integral shoulders 60, the same abutment effect could be achieved by sliding a sleeve over a constant cross section elongate member, the sleeve having a length approximating the distance between the inner forks 62.Rather than pass either of the alternate elongate members 50, 58 through the aperture 52 usually occupied by the wheel axles 51, a separate aperture 61 may be provided on each yoke 19 away from the wheels 22 as depicted in FIG.","9.The elongated rod 50, 58 is thus passed through the apertures 61 and secured to the yokes 19 in the manner discussed above.","The original axles 51 remain in place.","Other alternative forms of securing the elongate member 50, 58 to the frames will be appreciated by the person skilled in the art.","One such alternative form would be the use of circlips to replace the nuts 54 abutting the outer forks 63.It is preferred that the modified wheel assemblies to include a fixed wheel assembly in any form detailed above is installed on the front wheel assembly 12 as the provision of an elongate rod spanning the rear casters 17 or the presence of a centre fixed wheel may act as a barrier to nesting of trolleys one inside the other.","The elongate member or fixed wheel assembly may jamb against the front of a trolley being stacked into the back of a trolley so modified.","It is also preferred that the wheels 22 are positioned such that their rotation axes are co-axial, however slight misalignment, including a slight toe-in or toe-out of the wheels will still provide effective manoeuvrability.","It is to be appreciated that variations and/or modifications may be made to the parts previously described without departing from the spirit or ambit of the invention."]],"string":"[\n [\n \"Trolley wheel assembly\",\n \"A trolley includes a front wheel assemblie (12) and a rear wheel assembly both assemblies mounted on a trolley frame.\",\n \"The front wheel assembly (12) includes a fixed wheel assembly (24) to aid control of the trolley.\",\n \"In one form the front wheel assembly (12) includes a pair of swivel castors (18) and a fixed wheel assembly (24) disposed between the castors (18).\",\n \"The front wheel assembly (12) is designed so that when the castors (18) swivel from their orientation for forward travel, the fixed wheel assembly (24) is caused to be lifted off the ground.\",\n \"The trolley also includes a camming device which is operative to lift the front wheel assembly (12) off the ground when the trolley is moved into a nested configuration with a like trolley.\"\n ],\n [\n \"1.A trolley comprising a trolley frame, and front and rear wheel assemblies, one of the wheel assemblies including a pair of casters, each caster including a yoke mounted to the trolley frame in a manner that allows the caster to swivel about a first axis, and a caster wheel mounted to the yoke so as to be rotatable about a second axis which is transverse to said first axis, each caster being operative to adopt a first orientation for forward travel of said trolley, the one wheel assembly being configured so as to lift the trolley frame when the casters swivel about their first axes from the first orientation, wherein the one wheel assembly further comprises a fixed wheel assembly mounted to the trolley frame and including at least one wheel which has an axis of rotation which is orientated so that said fixed wheel assembly assists in maintaining directional control of said trolley in its forward travel, the wheel of the fixed wheel assembly being positioned to engage the ground when the castors are in the first orientation and to be off the ground when the trolley frame is lifted by swivelling of the casters about their first axes.\",\n \"2.A trolley according to claim 1, wherein the one wheel assembly is configured to lift the trolley frame on swivelling of the casters from the first orientation by having the centre of the caster wheels radially displaced from their first axes and having the first axes inclined to the vertical and extending rearwardly of the trolley.\",\n \"3.A trolley according to claim 1, wherein the fixed wheel assembly is disposed between the casters of the one wheel assembly.\",\n \"4.A trolley according to claim 1, the one wheel assembly is the front wheel assembly.\",\n \"5.A trolley according to claim 4, wherein the wheel of the fixed wheel assembly is aligned with the casters of the front wheel assembly.\",\n \"6.A trolley according to claim 4 wherein the fixed wheel assembly is displaced either forward or rearward of the casters of the front wheel assembly.\",\n \"7.A trolley according to claim 1, the other wheel assembly includes a pair of casters, each of the other wheel assembly casters including a yoke mounted to the trolley frame in a manner that allows the caster to swivel about a first axis, and a caster wheel mounted to the yoke so as to be rotatable about a second axis which is transverse to said first axis, each caster being operative to adopt a first orientation for forward travel of said trolley.\",\n \"8.A trolley according to claim 1, wherein the fixed wheel assembly is the form of a fixed caster including a yoke mounted to the trolley frame, and a caster wheel rotatably mounted to the yoke.\",\n \"9.A trolley comprising a trolley frame, and front and wheel assemblies, one of said wheel assemblies including a pair of casters each including a yoke mounted to the trolley frame which is operative to allow the casters to swivel about a first axis, and a caster wheel mounted to the yoke so as to be rotatable about a second axis which is transverse to said first axis, and an elongate member which extends through the respective yokes of said casters so as to maintain the casters in a first orientation for forward travel of said trolley and thereby prevent them swivelling about their respective first axes.\",\n \"10.A trolley according to claim 9, wherein the elongate member forms the axle of the respective casters and thereby supports the caster wheels for rotational movement.\",\n \"11.A trolley according to claim 9, wherein the trolley further comprises a fixed wheel assembly having at least one wheel and the elongate member forms the axle of the fixed wheel assembly and thereby supports the at least one wheel for rotational movement.\",\n \"12.A trolley according to claim 11, wherein the caster wheels are smaller than the at least one wheel of the fixed wheel assembly.\",\n \"13.A trolley according to claim 11, wherein the trolley is designed to be nested with another said trolley, wherein the front portion of one trolley is in overlapping relation with the rear portion of the other trolley, and wherein the trolley further includes lifting means which is operative to cause the wheels of the front wheel assembly of the one trolley to be lifted off the ground on nesting of the two trolleys.\",\n \"14.A trolley according to claim 13, wherein the lifting means is in the form of complementary surfaces arranged to interengage during nesting of the trolleys.\",\n \"15.A trolley according to claim 14, further comprising a camming device which includes a camming surface, the camming device being secured to the trolley so that the camming surface forms one of the complementary surfaces and engages a surface of the other trolley on nesting of the trolleys.\",\n \"16.A trolley according to claim 15, wherein the trolley includes a basket having a rear gate, and wherein on nesting, the basket of the one trolley locates in the basket of the other trolley through the rear gate, and wherein the camming surface is positioned so as to engage a surface of the basket to cause lifting of the one trolley.\",\n \"17.A trolley according to claim 16, wherein the camming device is mounted on a base surface of the basket and projects downwardly therefrom.\",\n \"18.A trolley according to claim 16, the camming device is mounted to a base of the basket and projects upwardly therefrom.\",\n \"19.A trolley according to claim 15, wherein the camming device comprises an elongate member having opposite ends and being profiled to form the camming surface intermediate the ends, the member further comprising engagement elements located at each end which are operative to be fixed to the basket of said trolley.\",\n \"20.A trolley according to claim 19, wherein the basket of the trolley is formed from wire mesh and the engagement elements are in the form of eyelets which are operative to engage respective wire bars of the trolley basket.\",\n \"21.A trolley comprising a trolley frame, and front and rear wheel assemblies mounted on said trolley frame, the trolley being designed to be nested with another trolley where a front portion of one trolley is in overlapping relation with a rear portion of the other trolley, and wherein the trolley further comprises lifting means operative to cause the wheels of the front wheel assembly of the one trolley to be lifted off the ground during nesting of the one trolley with the other trolley.\",\n \"22.A camming device for a trolley, the camming device comprising an elongate member having opposite ends and being profiled to form a camming surface intermediate the ends, the member further comprising engagement elements located at each end that are operative to be fixed to the trolley.\",\n \"23.A camming device according to claim 22, engagement elements are in the form of eyelets.\",\n \"24.A camming device according to claim 22, wherein the camming device is made from sheet steel.\",\n \"25.A method of retrofitting a trolley having front and rear wheel assemblies to incorporate a camming surface to improve nesting of the trolley with another trolley, the method comprising the steps of: providing a camming device comprising an elongate member having opposite ends and being profiled to form a camming surface intermediate the ends, and securing the camming device to the trolley in a position so that when the trolley is moved into a nested configuration with the other trolley, the camming surface is operative to engage the other trolley in the nested configuration so as to cause the rear trolley in the nested configuration to be lifted so that the front wheels of that trolley disengage the ground.\",\n \"26.A method of securing a pair of casters mounted on a trolley body against swivelling, each caster having a wheel mounted on an axle passing through a yoke which is in turn rotatably mounted on said trolley body, said method including the steps of: providing an aperture in the yoke of each of said casters; positioning each caster wheel such that the rotation axes of the wheels are substantially coaxial; passing a rigid elongate member through each said aperture so as to maintain said rotational axis in a substantially coaxial relationship; and fixedly securing said elongate members to said yoke.\",\n \"27.A pair of casters when secured against swivelling by a method according to claim 26.28.\",\n \"(canceled) 29.\",\n \"(canceled) 30.\",\n \"(canceled) 31.\",\n \"(canceled) 32.\",\n \"(canceled) 33.\",\n \"(canceled) 34.\",\n \"(canceled)\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>Shopping trolleys are typically configured with four fully swivelling casters, one caster being provided at each corner of the base of the trolley frame to form front and rear wheel assemblies.\",\n \"The use of four fully swivelling casters enables the shopping trolley to be manoeuvred forwards or sideways, however can render the trolley difficult to control, with steering a trolley around a corner or maintaining it in a straight path or on a sloped surface being particularly difficult.\",\n \"With the casters being free to swivel and therefore not providing any sideways resistance, altering the path of the trolley requires the operator to impart a moment directly on the trolley to rotate it as there are no fixed wheels about which the trolley can easily rotate.\",\n \"The difficulty in manoeuvring a trolley is increased when loaded with shopping, with the increased weight resulting in an increased moment being required to alter the path of the trolley.\",\n \"The applicant has found that a shopping trolley having the front pair of wheels fixed from swivelling, with the wheel axles in a co-axial relationship, and the rear wheels being able to swivel, not unlike a motor vehicle, is easier to manoeuvre.\",\n \"A trolley with such a configuration is able to pivot about the fixed set of wheels to alter its direction.\",\n \"The operator need only control this pivoting about the fixed wheels by applying a small sideways force to the trolley handle to direct the trolley along the required path.\",\n \"Whilst this arrangement is effective, the manoeuvrability of a trolley may be unduly limited, particularly when turning the trolley in tight situations.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>Accordingly, an aim of the present invention is to ameliorate this problem whilst still utilising the advantages to trolleys of including a fixed wheel assembly.\",\n \"In its broadest terms, the invention is directed to a trolley including a trolley frame, and front and rear wheel assemblies, one of said wheel assemblies including a pair of casters, each including a yoke mounted to the trolley frame in a manner that allows the caster to swivel about a first axis, and a caster wheel mounted to the yoke so as to be rotatable about a second axis which is transverse to said first axis, each caster being operative to adopt a first orientation for forward travel of said trolley, and a fixed wheel assembly located between said casters, said fixed wheel assembly including at least one wheel which has an axis of rotation which is orientated so that said fixed wheel assembly assists in maintaining directional control of said trolley in its forward travel.\",\n \"In one form, the wheels of fixed wheel assembly and the casters in their first orientation are designed to engage the ground.\",\n \"In this arrangement, the fixed wheel assembly provides the directional control for the trolley, whilst the casters which are able to swivel provide the lateral stability for the trolley to prevent tipping.\",\n \"In another form, the wheel of fixed wheel assembly is designed to project below the level of the caster wheels when in their first orientation so that, in forward travel, the trolley is supported at said one wheel assembly by the fixed wheel assembly.\",\n \"The casters which may be fixed or swivelling, act as outrigger wheels to prevent sideways tipping of the trolley.\",\n \"In this latter arrangement, preferably the fixed wheel assembly includes a plurality of closely spaced wheels to provide more balanced support at the front of the trolley.\",\n \"The arrangement of the invention has significant advantages.\",\n \"By providing a central fixed wheel, enables the trolley to be manoeuvred more easily than is the case if the fixed wheels are widely spaced apart.\",\n \"Further, it provides a system which can be easily and inexpensively retrofitted to existing trolleys.\",\n \"It also maintains a useful purpose for the existing casters to enable the trolley to be stably supported.\",\n \"In particular, the arrangement enables the trolley to meet some government requirements whereby a trolley must be supplied with at least four wheels.\",\n \"In a particularly preferred form, when each caster adopts a second orientation which is angularly displaced from the first orientation about the first axis, the wheels of said casters are operative to extend below said wheel of said fixed wheel assembly so as to be operative to cause the wheel of the fixed wheel assembly to be off the ground.\",\n \"Preferably the second orientation is angularly displaced from said first orientation by 90 degrees.\",\n \"More preferably, the fixed wheel is operative to be lifted off the ground when either caster is displaced angularly from the first orientation by more than 20 degrees.\",\n \"Preferably, the caster wheel in each caster is offset from its first axis so that, in use, the caster wheel swivels about an arc around its first axis.\",\n \"In addition, preferably the caster is mounted to the trolley frame with the first axis inclined to the vertical so that in use, once swivelling of the casters about their respective first axes, the vertical displacement of the centre of the respective casters from a fixed point on the trolley varies.\",\n \"In a particularly preferred form, each caster is mounted to the trolley so that this vertical displacement is at a minimum when that caster is in its first orientation.\",\n \"This arrangement has particular advantage in that when either of the casters is caused to be angularly spaced about its first axis from the first orientation by a predetermined amount, that caster wheel is lower than the fixed wheel thereby lifting the fixed wheel off the ground.\",\n \"The degree of angular displacement required to provide adequate clearance to fully lift the fixed wheel off the ground will depend on the specific configuration of the wheel assembly, but the applicant has found that displacement of 10-20 degrees from the first orientation is ideal.\",\n \"Further, if the casters are operative to remain in contact with the ground at all times, they can be caused to swivel from the first orientation (and thereby lift the fixed wheel off the ground) simply by applying a sideways force to the trolley.\",\n \"This arrangement has the advantage that the fixed wheel assembly assists in controlling forward movement of the trolley, but does not inhibit sideways movement of the trolley as the caster wheels, as they pivot, would be caused to lift this fixed wheel assembly off the ground.\",\n \"In addition, the fixed wheel can inhibit the trolley inadvertently “running away” when it is stationery as it lessens the tendency to move sideways if parked on a sideways sloping surface.\",\n \"In one arrangement, the novel wheel assembly may be retrofitted to an existing trolley assembly by securing a fixed wheel assembly to the trolley body.\",\n \"This may be secured to the existing cross bar supporting the caster wheel assemblies or an additional cross bar may be installed.\",\n \"Further, the casters may be arranged to have their axes inclined to the vertical in the desired orientation merely by tilting the trolley frame which can be simply achieved by placing larger casters on the rear wheel assembly.\",\n \"Alternatively the caster swivel axis may be inclined by the addition of an appropriately tapered spacers mounted at the point of attachment of the caster to the trolley.\",\n \"In another form, an elongate rigid member is used to interconnect the frames of the caster wheel assemblies.\",\n \"In this latter arrangement, preferably the elongate member is used as the axle for the fixed wheel assembly.\",\n \"Appropriate spaces, or fixtures are provided on the axle to properly locate those fixed wheels in place.\",\n \"In a particularly preferred form of this latter arrangement, the elongate member extends through the frame axles of the respective casters.\",\n \"Further, the caster wheels from the pair of assemblies are removed and are used as part of the fixed wheel assembly.\",\n \"In their place, smaller outrigger wheels are used.\",\n \"The advantage of this arrangement is that the caster wheels are designed to be constantly in contact with the ground.\",\n \"As such, they are ideally suited to be used as the fixed wheel assembly.\",\n \"Further as the outrigger wheels are only used to prevent tipping of the trolley, an inferior wheel may be used as compared to the caster wheels, thereby further reducing the cost of retrofitting.\",\n \"In yet a further aspect, the invention is directed to a method of retrofitting a trolley using any one of the techniques described above.\",\n \"In a particularly preferred form, the front wheel assembly includes the fixed wheel assembly.\",\n \"In this arrangement preferably the rear wheel assembly includes pivoting casters, although other arrangements may be used.\",\n \"Whilst the design of the shopping trolley with a fixed wheel as part of the front wheel assembly aids control of an individual trolley, it may impede the manoeuvrability of the trolleys when in a nested configuration as a train as it significantly restricts the ability to be able to change the direction of the train of trolleys.\",\n \"Accordingly to a further aspect, the present invention provides a trolley including a trolley frame, and front and rear wheel assemblies mounted on said trolley frame, the trolley being designed to be nested with a like trolley where a front portion of one said trolley is in overlapping relation with a rear portion of another said trolley, and wherein the trolley further includes lifting means which is operative to cause the wheels of the front wheel assembly of the one trolley to be lifted off the ground during nesting of the one trolley with said other trolley.\",\n \"A trolley according to the above form has significant advantage.\",\n \"The trolley may exhibit improved manoeuvrability by having a fixed wheel assembly at the front wheel assembly according to any form described above.\",\n \"Further, the manoeuvrability of a train of nested trolleys in accordance with the invention is also significantly easier to manoeuvre by virtue of the lifting means which disengages each of the front wheels assemblies of the nested trolleys off the ground.\",\n \"With this arrangement, only the front wheel assembly of the front trolley in the train is in engagement with the ground.\",\n \"Therefore the train includes one fixed front wheel assembly with the remaining wheels being part of the individual trolleys rear wheels which are preferably casters.\",\n \"This provides an arrangement which is easier to manoeuvre than traditional trains of nested shopping trolleys which includes only swivelling casters.\",\n \"Further, by virtue of the fact that the front wheel assemblies of the nested trolleys is disengaged from the ground, the number of wheels actually in contact with the ground in any train is significantly reduced which further improves the manoeuvrability of the train of nested trolleys.\",\n \"In a preferred form, the lifting means is in the form of a camming surface arranged to engage a complementary surface on the like trolley during nesting.\",\n \"Through that engagement, the wheels on the front wheel assembly of the one trolley is caused to move off the ground.\",\n \"This camming arrangement may be located on any suitable part of the trolley such as on the basket, or trolley frame.\",\n \"In an alternative form, the front wheel assembly may be operative to move out of engagement with the ground using a different mechanism as the lifting means.\",\n \"For example, the wheel assemblies may be pivotable from an extended to a retracted configuration, and wherein on nesting of the one trolley with the like trolley, the front wheel assemblies may be caused to move from their extended position to their retracted position.\",\n \"In yet a further aspect, the present invention relates to the retrofitting of existing shopping trolleys so as to provide the enhanced manoeuvrability afforded by the shopping trolleys of the earlier aspects of the invention.\",\n \"In particular, in one aspect the invention is directed to retrofitting existing shopping trolleys so as to incorporate the lifting means.\",\n \"This aspect aims to use techniques which are inexpensive which thereby will not be prohibitively expensive to retrofit hundreds of shopping trolleys in a typical supermarket.\",\n \"In accordance with this further aspect, the invention relates to a camming device for a trolley, the camming device including a body incorporating a camming surface, and attachment means operative to fix the camming device to said shopping trolley, the trolley including a basket incorporating a rear gate and the trolley being designed so as to be able to adopt a nested configuration with a like trolley, where the basket of one trolley locate through the rear gate of another trolley so that when nested, a substantial portion of the basket of the one trolley is disposed within the basket of the like trolley, and wherein in use, when the camming device is fixed to the trolley, the camming surface is operative to engage the other trolley in the nested configuration so as to lift the one trolley as it is moved into the nested configuration so as to cause the front wheel assembly to disengage the ground.\",\n \"In a preferred form, the camming device includes an elongate member having opposite ends, the member being shaped so as to form a camming surface intermediate the ends, and wherein the attachment means comprises engagement elements located at each end which are operative to be fixed to the trolley.\",\n \"In a preferred form, the camming device is operative to be secured to the basket of the trolley and the engagement means is preferably in the form of eyelets which are operative to engage respective wire bars of the trolley basket.\",\n \"The advantage of this arrangement is that the camming device is inexpensive, and the method of attachment can be easily performed by providing the eyelets in partially open form on respective wire bars of the basket and merely clinching them to securely affix them in place.\",\n \"In yet a further aspect, the present invention relates to a method of retrofitting a trolley to incorporate a camming device which includes the steps of providing a camming device in any form described above, and securing the camming device to the trolley in a position so that when the trolley is moved into a nested configuration with a like trolley, the camming device on one of the trolleys is operative to engage the other of the trolley in the nested configuration so as to cause the one trolley to be lifted so that the front wheels disengage the ground in the nested configuration.\",\n \"It is to be appreciated that the trolley including the lifting means in accordance with these aspects of the invention is ideally suited to be used in conjunction with a trolley including the front wheel assembly with a fixed wheel assembly in accordance with the earlier aspects of the invention.\",\n \"Alternatively, it may be used with trolleys having other arrangements of front wheel assemblies, whether they include a fixed wheel or not.\",\n \"In yet a further aspect, the invention relates to methods and systems to modify an existing pair of caster wheel assemblies so as to provide a fixed axle wheel arrangement using simple and inexpensive techniques.\",\n \"Accordingly, in this aspect, there is disclosed herein apparatus adapted to secure a pair of casters mounted on a trolley body against swivelling, each said caster having a wheel mounted on an axle passing through a yoke which is in turn rotatably mounted on said trolley body, said apparatus including: a rigid elongate member adapted to pass through an aperture provided in each said yoke such that the rotation axes of said wheels are substantially coaxial, and means for fixedly securing said elongate member to said yokes.\",\n \"Typically said elongate member has a circular cross-section toward each end therefor and is adapted to replace the axle of each of said casters.\",\n \"Typically said securing means includes a pair of nuts each respectively adapted to thread onto a threaded portion provided at each respective end of said elongate member so as to abut the respective said yoke.\",\n \"Typically said securing means includes means for abutting each said yoke on an opposing side thereof to the associated elongate member end.\",\n \"Preferably said abutting means includes a pair of circlips each adapted to be received in a groove provided in said elongate member, each said groove being positioned such that, in use, each said circlip abuts a said frame.\",\n \"Preferably said abutting means includes a pair of shoulders each defined by a change in cross-section of said elongate member.\",\n \"Alternatively the apparatus further includes a sleeve locatable over the elongate member and wherein opposite ends of the sleeve provides said abutting means.\",\n \"There is further disclosed herein a method of securing a pair of casters mounted on a trolley body against swivelling, each said caster having a wheel mounted on an axle passing through a yoke which is in turn rotatably mounted on said trolley body, said method including the steps of: providing an aperture in the yoke of each of said caster wheel assemblies; positioning each said caster such that the rotation axes of said wheels are substantially co-axial; passing a rigid elongate member through each said aperture so as to maintain said rotation axes in a substantially co-axial relationship, and fixedly securing said elongate member to said yokes.\",\n \"Typically said elongate member is circular in cross-section towards each end thereof and said step of providing an aperture in each yoke is achieved by removing the axle of each said caster, said elongate member replacing said axles so as to provide said axes of rotation of said wheels.\",\n \"There is still further disclosed herein a method of modifying the handling characteristics of a shopping trolley having four caster wheel assemblies mounted on the body thereof, said method including the step of securing either the front or rear pair of caster wheels against swivelling according to the above disclosed method.\",\n \"There is still further disclosed herein a trolley modified by the above method.\",\n \"Accordingly, the present invention provides a novel trolley and associated methods of modification of existing trolleys, which has the advantage of the fixed front wheels, but have improved manoeuvrability, and which may be inexpensively retrofitted to be incorporated into existing trolleys.\"\n ],\n [\n \"TECHNICAL FIELD The present invention relates to a trolley and in particular to a trolley with a modified wheel assembly to aid in control of the trolley.\",\n \"The invention has been designed especially, but not exclusively, for use in shopping trolleys and the invention is herein described in that context.\",\n \"It is to be appreciated however that the invention is not limited to that use.\",\n \"BACKGROUND OF THE INVENTION Shopping trolleys are typically configured with four fully swivelling casters, one caster being provided at each corner of the base of the trolley frame to form front and rear wheel assemblies.\",\n \"The use of four fully swivelling casters enables the shopping trolley to be manoeuvred forwards or sideways, however can render the trolley difficult to control, with steering a trolley around a corner or maintaining it in a straight path or on a sloped surface being particularly difficult.\",\n \"With the casters being free to swivel and therefore not providing any sideways resistance, altering the path of the trolley requires the operator to impart a moment directly on the trolley to rotate it as there are no fixed wheels about which the trolley can easily rotate.\",\n \"The difficulty in manoeuvring a trolley is increased when loaded with shopping, with the increased weight resulting in an increased moment being required to alter the path of the trolley.\",\n \"The applicant has found that a shopping trolley having the front pair of wheels fixed from swivelling, with the wheel axles in a co-axial relationship, and the rear wheels being able to swivel, not unlike a motor vehicle, is easier to manoeuvre.\",\n \"A trolley with such a configuration is able to pivot about the fixed set of wheels to alter its direction.\",\n \"The operator need only control this pivoting about the fixed wheels by applying a small sideways force to the trolley handle to direct the trolley along the required path.\",\n \"Whilst this arrangement is effective, the manoeuvrability of a trolley may be unduly limited, particularly when turning the trolley in tight situations.\",\n \"SUMMARY OF THE INVENTION Accordingly, an aim of the present invention is to ameliorate this problem whilst still utilising the advantages to trolleys of including a fixed wheel assembly.\",\n \"In its broadest terms, the invention is directed to a trolley including a trolley frame, and front and rear wheel assemblies, one of said wheel assemblies including a pair of casters, each including a yoke mounted to the trolley frame in a manner that allows the caster to swivel about a first axis, and a caster wheel mounted to the yoke so as to be rotatable about a second axis which is transverse to said first axis, each caster being operative to adopt a first orientation for forward travel of said trolley, and a fixed wheel assembly located between said casters, said fixed wheel assembly including at least one wheel which has an axis of rotation which is orientated so that said fixed wheel assembly assists in maintaining directional control of said trolley in its forward travel.\",\n \"In one form, the wheels of fixed wheel assembly and the casters in their first orientation are designed to engage the ground.\",\n \"In this arrangement, the fixed wheel assembly provides the directional control for the trolley, whilst the casters which are able to swivel provide the lateral stability for the trolley to prevent tipping.\",\n \"In another form, the wheel of fixed wheel assembly is designed to project below the level of the caster wheels when in their first orientation so that, in forward travel, the trolley is supported at said one wheel assembly by the fixed wheel assembly.\",\n \"The casters which may be fixed or swivelling, act as outrigger wheels to prevent sideways tipping of the trolley.\",\n \"In this latter arrangement, preferably the fixed wheel assembly includes a plurality of closely spaced wheels to provide more balanced support at the front of the trolley.\",\n \"The arrangement of the invention has significant advantages.\",\n \"By providing a central fixed wheel, enables the trolley to be manoeuvred more easily than is the case if the fixed wheels are widely spaced apart.\",\n \"Further, it provides a system which can be easily and inexpensively retrofitted to existing trolleys.\",\n \"It also maintains a useful purpose for the existing casters to enable the trolley to be stably supported.\",\n \"In particular, the arrangement enables the trolley to meet some government requirements whereby a trolley must be supplied with at least four wheels.\",\n \"In a particularly preferred form, when each caster adopts a second orientation which is angularly displaced from the first orientation about the first axis, the wheels of said casters are operative to extend below said wheel of said fixed wheel assembly so as to be operative to cause the wheel of the fixed wheel assembly to be off the ground.\",\n \"Preferably the second orientation is angularly displaced from said first orientation by 90 degrees.\",\n \"More preferably, the fixed wheel is operative to be lifted off the ground when either caster is displaced angularly from the first orientation by more than 20 degrees.\",\n \"Preferably, the caster wheel in each caster is offset from its first axis so that, in use, the caster wheel swivels about an arc around its first axis.\",\n \"In addition, preferably the caster is mounted to the trolley frame with the first axis inclined to the vertical so that in use, once swivelling of the casters about their respective first axes, the vertical displacement of the centre of the respective casters from a fixed point on the trolley varies.\",\n \"In a particularly preferred form, each caster is mounted to the trolley so that this vertical displacement is at a minimum when that caster is in its first orientation.\",\n \"This arrangement has particular advantage in that when either of the casters is caused to be angularly spaced about its first axis from the first orientation by a predetermined amount, that caster wheel is lower than the fixed wheel thereby lifting the fixed wheel off the ground.\",\n \"The degree of angular displacement required to provide adequate clearance to fully lift the fixed wheel off the ground will depend on the specific configuration of the wheel assembly, but the applicant has found that displacement of 10-20 degrees from the first orientation is ideal.\",\n \"Further, if the casters are operative to remain in contact with the ground at all times, they can be caused to swivel from the first orientation (and thereby lift the fixed wheel off the ground) simply by applying a sideways force to the trolley.\",\n \"This arrangement has the advantage that the fixed wheel assembly assists in controlling forward movement of the trolley, but does not inhibit sideways movement of the trolley as the caster wheels, as they pivot, would be caused to lift this fixed wheel assembly off the ground.\",\n \"In addition, the fixed wheel can inhibit the trolley inadvertently “running away” when it is stationery as it lessens the tendency to move sideways if parked on a sideways sloping surface.\",\n \"In one arrangement, the novel wheel assembly may be retrofitted to an existing trolley assembly by securing a fixed wheel assembly to the trolley body.\",\n \"This may be secured to the existing cross bar supporting the caster wheel assemblies or an additional cross bar may be installed.\",\n \"Further, the casters may be arranged to have their axes inclined to the vertical in the desired orientation merely by tilting the trolley frame which can be simply achieved by placing larger casters on the rear wheel assembly.\",\n \"Alternatively the caster swivel axis may be inclined by the addition of an appropriately tapered spacers mounted at the point of attachment of the caster to the trolley.\",\n \"In another form, an elongate rigid member is used to interconnect the frames of the caster wheel assemblies.\",\n \"In this latter arrangement, preferably the elongate member is used as the axle for the fixed wheel assembly.\",\n \"Appropriate spaces, or fixtures are provided on the axle to properly locate those fixed wheels in place.\",\n \"In a particularly preferred form of this latter arrangement, the elongate member extends through the frame axles of the respective casters.\",\n \"Further, the caster wheels from the pair of assemblies are removed and are used as part of the fixed wheel assembly.\",\n \"In their place, smaller outrigger wheels are used.\",\n \"The advantage of this arrangement is that the caster wheels are designed to be constantly in contact with the ground.\",\n \"As such, they are ideally suited to be used as the fixed wheel assembly.\",\n \"Further as the outrigger wheels are only used to prevent tipping of the trolley, an inferior wheel may be used as compared to the caster wheels, thereby further reducing the cost of retrofitting.\",\n \"In yet a further aspect, the invention is directed to a method of retrofitting a trolley using any one of the techniques described above.\",\n \"In a particularly preferred form, the front wheel assembly includes the fixed wheel assembly.\",\n \"In this arrangement preferably the rear wheel assembly includes pivoting casters, although other arrangements may be used.\",\n \"Whilst the design of the shopping trolley with a fixed wheel as part of the front wheel assembly aids control of an individual trolley, it may impede the manoeuvrability of the trolleys when in a nested configuration as a train as it significantly restricts the ability to be able to change the direction of the train of trolleys.\",\n \"Accordingly to a further aspect, the present invention provides a trolley including a trolley frame, and front and rear wheel assemblies mounted on said trolley frame, the trolley being designed to be nested with a like trolley where a front portion of one said trolley is in overlapping relation with a rear portion of another said trolley, and wherein the trolley further includes lifting means which is operative to cause the wheels of the front wheel assembly of the one trolley to be lifted off the ground during nesting of the one trolley with said other trolley.\",\n \"A trolley according to the above form has significant advantage.\",\n \"The trolley may exhibit improved manoeuvrability by having a fixed wheel assembly at the front wheel assembly according to any form described above.\",\n \"Further, the manoeuvrability of a train of nested trolleys in accordance with the invention is also significantly easier to manoeuvre by virtue of the lifting means which disengages each of the front wheels assemblies of the nested trolleys off the ground.\",\n \"With this arrangement, only the front wheel assembly of the front trolley in the train is in engagement with the ground.\",\n \"Therefore the train includes one fixed front wheel assembly with the remaining wheels being part of the individual trolleys rear wheels which are preferably casters.\",\n \"This provides an arrangement which is easier to manoeuvre than traditional trains of nested shopping trolleys which includes only swivelling casters.\",\n \"Further, by virtue of the fact that the front wheel assemblies of the nested trolleys is disengaged from the ground, the number of wheels actually in contact with the ground in any train is significantly reduced which further improves the manoeuvrability of the train of nested trolleys.\",\n \"In a preferred form, the lifting means is in the form of a camming surface arranged to engage a complementary surface on the like trolley during nesting.\",\n \"Through that engagement, the wheels on the front wheel assembly of the one trolley is caused to move off the ground.\",\n \"This camming arrangement may be located on any suitable part of the trolley such as on the basket, or trolley frame.\",\n \"In an alternative form, the front wheel assembly may be operative to move out of engagement with the ground using a different mechanism as the lifting means.\",\n \"For example, the wheel assemblies may be pivotable from an extended to a retracted configuration, and wherein on nesting of the one trolley with the like trolley, the front wheel assemblies may be caused to move from their extended position to their retracted position.\",\n \"In yet a further aspect, the present invention relates to the retrofitting of existing shopping trolleys so as to provide the enhanced manoeuvrability afforded by the shopping trolleys of the earlier aspects of the invention.\",\n \"In particular, in one aspect the invention is directed to retrofitting existing shopping trolleys so as to incorporate the lifting means.\",\n \"This aspect aims to use techniques which are inexpensive which thereby will not be prohibitively expensive to retrofit hundreds of shopping trolleys in a typical supermarket.\",\n \"In accordance with this further aspect, the invention relates to a camming device for a trolley, the camming device including a body incorporating a camming surface, and attachment means operative to fix the camming device to said shopping trolley, the trolley including a basket incorporating a rear gate and the trolley being designed so as to be able to adopt a nested configuration with a like trolley, where the basket of one trolley locate through the rear gate of another trolley so that when nested, a substantial portion of the basket of the one trolley is disposed within the basket of the like trolley, and wherein in use, when the camming device is fixed to the trolley, the camming surface is operative to engage the other trolley in the nested configuration so as to lift the one trolley as it is moved into the nested configuration so as to cause the front wheel assembly to disengage the ground.\",\n \"In a preferred form, the camming device includes an elongate member having opposite ends, the member being shaped so as to form a camming surface intermediate the ends, and wherein the attachment means comprises engagement elements located at each end which are operative to be fixed to the trolley.\",\n \"In a preferred form, the camming device is operative to be secured to the basket of the trolley and the engagement means is preferably in the form of eyelets which are operative to engage respective wire bars of the trolley basket.\",\n \"The advantage of this arrangement is that the camming device is inexpensive, and the method of attachment can be easily performed by providing the eyelets in partially open form on respective wire bars of the basket and merely clinching them to securely affix them in place.\",\n \"In yet a further aspect, the present invention relates to a method of retrofitting a trolley to incorporate a camming device which includes the steps of providing a camming device in any form described above, and securing the camming device to the trolley in a position so that when the trolley is moved into a nested configuration with a like trolley, the camming device on one of the trolleys is operative to engage the other of the trolley in the nested configuration so as to cause the one trolley to be lifted so that the front wheels disengage the ground in the nested configuration.\",\n \"It is to be appreciated that the trolley including the lifting means in accordance with these aspects of the invention is ideally suited to be used in conjunction with a trolley including the front wheel assembly with a fixed wheel assembly in accordance with the earlier aspects of the invention.\",\n \"Alternatively, it may be used with trolleys having other arrangements of front wheel assemblies, whether they include a fixed wheel or not.\",\n \"In yet a further aspect, the invention relates to methods and systems to modify an existing pair of caster wheel assemblies so as to provide a fixed axle wheel arrangement using simple and inexpensive techniques.\",\n \"Accordingly, in this aspect, there is disclosed herein apparatus adapted to secure a pair of casters mounted on a trolley body against swivelling, each said caster having a wheel mounted on an axle passing through a yoke which is in turn rotatably mounted on said trolley body, said apparatus including: a rigid elongate member adapted to pass through an aperture provided in each said yoke such that the rotation axes of said wheels are substantially coaxial, and means for fixedly securing said elongate member to said yokes.\",\n \"Typically said elongate member has a circular cross-section toward each end therefor and is adapted to replace the axle of each of said casters.\",\n \"Typically said securing means includes a pair of nuts each respectively adapted to thread onto a threaded portion provided at each respective end of said elongate member so as to abut the respective said yoke.\",\n \"Typically said securing means includes means for abutting each said yoke on an opposing side thereof to the associated elongate member end.\",\n \"Preferably said abutting means includes a pair of circlips each adapted to be received in a groove provided in said elongate member, each said groove being positioned such that, in use, each said circlip abuts a said frame.\",\n \"Preferably said abutting means includes a pair of shoulders each defined by a change in cross-section of said elongate member.\",\n \"Alternatively the apparatus further includes a sleeve locatable over the elongate member and wherein opposite ends of the sleeve provides said abutting means.\",\n \"There is further disclosed herein a method of securing a pair of casters mounted on a trolley body against swivelling, each said caster having a wheel mounted on an axle passing through a yoke which is in turn rotatably mounted on said trolley body, said method including the steps of: providing an aperture in the yoke of each of said caster wheel assemblies; positioning each said caster such that the rotation axes of said wheels are substantially co-axial; passing a rigid elongate member through each said aperture so as to maintain said rotation axes in a substantially co-axial relationship, and fixedly securing said elongate member to said yokes.\",\n \"Typically said elongate member is circular in cross-section towards each end thereof and said step of providing an aperture in each yoke is achieved by removing the axle of each said caster, said elongate member replacing said axles so as to provide said axes of rotation of said wheels.\",\n \"There is still further disclosed herein a method of modifying the handling characteristics of a shopping trolley having four caster wheel assemblies mounted on the body thereof, said method including the step of securing either the front or rear pair of caster wheels against swivelling according to the above disclosed method.\",\n \"There is still further disclosed herein a trolley modified by the above method.\",\n \"Accordingly, the present invention provides a novel trolley and associated methods of modification of existing trolleys, which has the advantage of the fixed front wheels, but have improved manoeuvrability, and which may be inexpensively retrofitted to be incorporated into existing trolleys.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS It is convenient to hereinafter describe embodiments of the present invention with reference to the accompanying drawings.\",\n \"It is to be appreciated that the particularity of the drawings and the related description is to be understood as not superseding the generality of the preceding broad description of the invention.\",\n \"In the drawings: FIG.\",\n \"1 is a side view of a shopping trolley according to a first embodiment of the invention; FIG.\",\n \"2 is a front elevation view of a front wheel assembly of the trolley of FIG.\",\n \"1; FIGS.\",\n \"3A and 3B are detailed side elevations of the front wheel assembly of FIG.\",\n \"2; FIG.\",\n \"4 is a front elevation view of a front wheel assembly for a trolley according to a second embodiment of the invention; FIG.\",\n \"5 is a detailed view of the camming device in the trolley of FIG.\",\n \"1; FIG.\",\n \"6 is a side view of the trolley of FIG.\",\n \"1 in a nested configuration; FIG.\",\n \"7 is a front elevation view of a pair of casters mounted on a trolley body and modified with the apparatus of a further embodiment of the invention; FIG.\",\n \"8 is a side elevation view of the casters of FIG.\",\n \"7; FIG.\",\n \"9 is a side elevation view of a pair of casters modified in an alternative manner; FIG.\",\n \"10 is a front elevation view of an elongate rod of the apparatus of FIG.\",\n \"7; and FIG.\",\n \"11 is a front elevation view of an elongate rod of the apparatus of FIG.\",\n \"9.DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS A shopping trolley 10 includes a frame 11 from which the front wheel assembly 12 and a rear wheel assembly 13 is mounted.\",\n \"A wire basket 14 is supported on the frame and includes a handle 15 and rear gate 16.As is typical in shopping trolleys, the rear wheel assembly includes a pair of swivel casters 17 mounted at each back corner of the frame 11.As best illustrated in FIGS.\",\n \"2, 3A and 3B, the front wheel assembly 12 include a pair of swivel casters which are mounted on respective front corners of the frame 11.Each caster 18 includes a yoke 19 which is arranged to rotate about a king pin 20 allowing the casters 18 to rotate about respective first axes 21.Caster wheels 22 are mounted on the respective yokes 19 and are rotatable about respective second axes 23 which are transverse to their respective first axes 21.To provide control to the trolley, the front wheel assembly 12 include a centre fixed wheel assembly 24 which is mounted to the trolley frame 11.In the illustrated form, the fixed wheel assembly 24 is displaced rearwardly of the front wheel assembly and is secured by a bracket 25 which is secured to the frame 11.In the illustrated form, the fixed wheel assembly 24 is secured to the bracket 25 by locating a threaded bolt 26 through an aperture in the bracket 25 which is secured in place by a nut 27.It is to be appreciated that other forms of fixing could be employed as will be appreciated by those skilled in the art.\",\n \"Further, the fixed wheel assembly 24 may be located in line with the casters 18 if there is adequate clearance to allow those casters to swivel, or disposed slightly forward.\",\n \"The fixed wheel assembly 24 is in the form of a non swivelling caster and includes a yoke 28 and a wheel 29 which is mounted to the yoke 28 and rotatable about an axis of rotation 30.FIGS.\",\n \"3A and 3B are a side elevation of the front wheel assembly of FIG.\",\n \"1.As can be seen, in this arrangement, the fixed wheel assembly 24 is secured rearward of the line of the casters 18.In addition, the first axes 21 of the casters 18 are each inclined to the vertical (V).\",\n \"This is achieved in the illustrated embodiment by having the trolley frame 11 slightly inclined forwardly through having larger casters on the rear wheel assembly than the front wheel assembly.\",\n \"Further, as illustrated in FIG.\",\n \"3A, the centre line (P) of each wheel 22 is displaced from its first axis 21.As such, the caster wheels 22 are designed to move in an arc around their first axes 21.Because each first axis 21 is inclined to the vertical, the plane in which the caster wheels will move on swivelling is inclined to the horizontal plane.\",\n \"In the illustrated embodiment, the first axes 21 of the casters 18 are inclined in the forward direction of the trolley.\",\n \"As a result of this orientation, when the casters 18 are in a trailing position for forward movement of the trolley as illustrated in FIG.\",\n \"3A, the caster wheels 22 are at the highest point of their arc.\",\n \"Further, the fixed wheel 29 of the trolley is designed so that it is aligned with the height of the caster wheels at this point so that both the caster wheels 18 and the fixed wheel 29 are engaging with the surface 200 on forward movement of the trolley.\",\n \"When the caster wheels 100 swivel about their first axes 21 away from the trailing position as shown in FIG.\",\n \"3A, the distance between the caster wheel centres and the trolley frame 11 increases, which causes the trolley frame 11 to lift.\",\n \"This, in turn, causes the fixed wheel 29 to move off the ground as illustrated in FIG.\",\n \"3B.\",\n \"This arrangement has significant practical benefit as it enables the fixed wheel 30 to assist in fully maintaining control of the trolley in the forward direction but still enables easy sideways movement of the trolley if required.\",\n \"Accordingly, front wheel assembly 12 has the advantages of a centralised fixed wheel for pivoting and manoeuvrability of the trolley particularly for forward movement, but which still can provide adequate stability to the trolley to prevent tipping.\",\n \"Further, the design of the front wheel assembly allows unimpeded sideways movement by creating a system where the fixed wheel can disengage the ground on certain orientations of the front casters.\",\n \"FIG.\",\n \"4 illustrates a modified version of the front wheel assembly 12.Again the main principal of the wheel assembly 12 is to provide a centralised fixed wheel between the front casters 18 to facilitate manoeuvrability of the trolley.\",\n \"In this arrangement however, the casters 18 are modified as part of the installation of the fixed wheel assembly 31.In particular in the arrangement of FIG.\",\n \"4, the caster wheels 22 are removed from their respective yoke 19 and those yokes are secured against swivelling by a rigid elongate member 32.Outrigger wheels 34 are included in the modified assembly 12 and the caster wheels 22 form part of the fixed centre wheel assembly 31.In constructing the front wheel assembly 12, an aperture is provided in each of the yokes 19 through which the elongate member 32 may pass when the casters 18 are positioned with the second axis 23 substantially coaxial.\",\n \"Individual axle in each caster 18 is removed so that the elongate member is passed through the axle apertures 35 provided in the respective yokes 19.In the illustrated form, the elongate member 32 includes a threaded portion 36 at each opposite end for receiving a nut 37 to secure the elongate member 32 in place.\",\n \"In conjunction with installing the elongate member 32 to actually fix the yokes 19, the caster wheels 22 are removed from the yokes 19 and are mounted on the member 32 intermediate the casters 18.Spaces 38 which in the illustrated form are sleeves of metal or plastic, are disposed on the elongate member to locate the caster wheels 22 in their central position.\",\n \"In this way, the elongate member which has a circular cross section provides the axis of rotation for the wheels 22 which are now centrally fixed.\",\n \"It is to be appreciated that other suitable spaces, such as circlips or the like may be used.\",\n \"In addition, outrigger wheels 34 are installed in the yokes 19 to provide additional stability to the trolley.\",\n \"These outrigger wheels are smaller than the casters 22 and are therefore only designed to come in contact with the ground should the trolley 10 tip.\",\n \"The trolley 10 in any form described above is designed to nest with a like trolley 10l as best shown in FIG.\",\n \"6.Specifically the trolley basket 14 with the pivoting rear gate 16, tapers towards its front.\",\n \"With this arrangement, the trolley 10 is able to nest with the like trolley 101 with the basket 14 locating through the rear gate 16l of the like trolley 10l so that when nested, the basket 14 of the trolley is disposed within the basket 14l of the like trolley.\",\n \"The trolley 10 includes a camming device 40 which is mounted to the basket 16.The device 40 includes an elongate metal member 41 which is arcuately shaped so as to form a cam surface 42 over its mid region.\",\n \"Each end (44,46) of the member 41 includes rolled over edges to form eyelets 45, 46.Wire bars 47 of the basket 16 are operative to locate within the eyelets 45, 46 so as to secure the camming device to the basket.\",\n \"To retrofit the camming device to the shopping trolley 10, the member 41 is provided with the eyelets partially opened so as to enable respective bars 47 of the basket 14 to be received in the eyelets whereafter they are secured in place by clinching the respective eyelets to close them.\",\n \"In fitting the camming device 40 in place, the shopping trolley 10 may be provided with a single device, or multiple devices may be provided.\",\n \"Further, in the illustrated form, the camming devices 40 depends downwardly from the basket 14.The advantage of this arrangement is that it places the camming device out of the way and does not reduce the volume of the material which can be provided within the trolley basket 14.Further, the formation of eyelets 45,46 at the end of the member provides a convenient safety edge to the camming device 20 which is unlikely to provide a pinch point or sharp edge which could otherwise catch on material placed in the basket 14.The inclusion of the camming device 40 is designed to cause a shopping trolley 10l when moved into a nested configuration to lift to a sufficient degree to cause the front wheel assembly 12l of nested trolley 10l to disengage the ground.\",\n \"This is achieved by the camming surface 22 moving into engagement with the bottom surface 481 of the trolley basket 141.By virtue of this engagement of surfaces, the trolley 10l is caused to lift disengaging the front wheel assemblies 12l from the ground.\",\n \"It is to be appreciated that the camming device 40 could be disposed to extend upwardly from the base of the trolley basket 14 and achieve the same result when a trolley is moved into a nesting configuration.\",\n \"When in this nested configuration, the lifting of the front wheel assemblies 12l ensures that these assemblies do not impede the manoeuvrability of a train of the trolleys in a nested configuration.\",\n \"FIGS.\",\n \"7 to 11 disclose yet a further form of modifying the casters of the front wheel assembly 12.This embodiment has many similarities to the embodiment shown with reference to FIG.\",\n \"4 with the exception that the modified trolley does not include a central fixed wheel assembly.\",\n \"As best shown in FIG.\",\n \"7, the wheels 22 of the casters 18 are secured against swivelling by a rigid elongate member 50.An aperture is provided in each of the yokes 19 through which the elongate member 50 may pass when the casters 18 are positioned with the rotation axes of the wheels 22 substantially co-axial.\",\n \"In one embodiment, the axles 51 of each wheel are removed so that the elongate member is passed through the axle apertures 52 provided in the yokes 19.The elongate member 50, whose ends have a circular cross-section, thus replaces the axles so as to provide the axes of rotation of the wheels 22, ensuring that the axes of rotation are co-axial.\",\n \"The elongate member 50 is then fixedly secured to the yokes 19.The elongate member 50 of this embodiment, which is in the form of a circular rod, is best depicted in FIG.\",\n \"10.A threaded portion 53 is provided at each opposing end of the elongate rod 50 for receiving a nut 54.A nut 54 is threaded onto each threaded portion 53 once the elongate rod has passed through the apertures 52.Each nut 54 is tightened onto the threaded portion 53 until it abuts the associated yoke 19 adjacent the elongate frame end.\",\n \"A washer 55 is typically provided between the nut 54 and yoke 19.Fastening the nuts 54 onto each end of the elongate rod 50 thus fixedly secures the elongate rod 50, with each end secured to its respective yoke 19.The rotation axes of the wheels 22 are thus fixed in the substantially co-axial relationship.\",\n \"To further secure the elongate rod 50, means may be provided to abut each yoke 19 on an opposing side thereof to the associated elongate member end.\",\n \"In the first embodiment this means is provided by circlips 56.The circlips 56 each abut an inner fork 62 of yoke 19, whilst a nut 54 abuts an outer fork 63 on the opposing side of the yoke 19.With each yoke 19 being abutted on opposing sides, the securing thereof is more robust.\",\n \"Each circlip 56 is received in a circumferential groove 57 provided in the elongate member 50 at a position adjacent to the inner fork 62 of each yoke 19 when the elongate member 50 is in position.\",\n \"An alternative elongate member 58 is depicted in FIG.\",\n \"11.Like elongate member 50 it is provided with threaded portions 59 for receiving nuts 57.However, in place of the grooves 57 for receiving circlips, shoulders 60 are provided at positions corresponding to adjacent the inner fork 62 of each yoke 19.The shoulders 60 are formed by a change in cross-section of the elongate member 58 from a larger cross-section in a central region to smaller cross-sections at outer regions of the elongate member.\",\n \"These shoulders 60 thus abut the inner forks 62 when the elongate member 58 is in position, helping to further secure the yokes 19 against swivelling.\",\n \"Rather than providing integral shoulders 60, the same abutment effect could be achieved by sliding a sleeve over a constant cross section elongate member, the sleeve having a length approximating the distance between the inner forks 62.Rather than pass either of the alternate elongate members 50, 58 through the aperture 52 usually occupied by the wheel axles 51, a separate aperture 61 may be provided on each yoke 19 away from the wheels 22 as depicted in FIG.\",\n \"9.The elongated rod 50, 58 is thus passed through the apertures 61 and secured to the yokes 19 in the manner discussed above.\",\n \"The original axles 51 remain in place.\",\n \"Other alternative forms of securing the elongate member 50, 58 to the frames will be appreciated by the person skilled in the art.\",\n \"One such alternative form would be the use of circlips to replace the nuts 54 abutting the outer forks 63.It is preferred that the modified wheel assemblies to include a fixed wheel assembly in any form detailed above is installed on the front wheel assembly 12 as the provision of an elongate rod spanning the rear casters 17 or the presence of a centre fixed wheel may act as a barrier to nesting of trolleys one inside the other.\",\n \"The elongate member or fixed wheel assembly may jamb against the front of a trolley being stacked into the back of a trolley so modified.\",\n \"It is also preferred that the wheels 22 are positioned such that their rotation axes are co-axial, however slight misalignment, including a slight toe-in or toe-out of the wheels will still provide effective manoeuvrability.\",\n \"It is to be appreciated that variations and/or modifications may be made to the parts previously described without departing from the spirit or ambit of the invention.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10474216"}}},{"rowIdx":790,"cells":{"text":{"kind":"list like","value":[["Method for obtaining data relating to the sentiment on a stock exchange","The present invention relates to a method for obtaining data relating to the sentiment of transactions on an (electronic) stock exchange, comprising the following steps of:—entering by a potential principal for stock exchange transactions electronically to one or more computers with one or more memories for storing therein one or more databases of transactions considered by this potential principal for a determined time period; and subsequently, i.e.","after a predetermined closing time for entering the transactions, determining a stock exchange sentiment on the basis of the content of the databases."],["1.A method for obtaining data relating to the sentiment of transactions on an (electronic) stock exchange, comprising the steps of: entering by a potential principal for stock exchange transactions electronically to one or more memories for storing therein one or more databases of transactions considered by this potential principal for a determined time period; and subsequently, i.e.","after a predetermined closing time for entering the transactions, determining a stock exchange sentiment on the basis of the content of the databases.","2.The method as claimed in claim 1, further comprising a step of inputting by the principal of data concerning the actually performed transactions.","3.The method as claimed in claim 1, further comprising a step for determining differences between intended transactions and performed transactions.","4.The method as claimed in claim 1, further comprising a step for inputting reasons for differences between intended transactions and performed transactions.","5.The method as claimed in claim 1, wherein the period of time is a day.","6.The method as claimed in claim 1, comprising a step for: determining a fund score in respect of a stock exchange fund by relating data concerning desired transactions in a period of time to data concerning desired transactions in a subsequent period, wherein the closing price of the day is stored in the database if it is a significant sentiment.","7.The method as claimed in claim 6, wherein a sentiment is significant if there is a difference in fund score of at least a predetermined percentage.","8.The method as claimed in claim 1, further comprising a step for determining a price prediction by relating the closing price from the databank in respect of desired transactions of the day to the closing price in respect of desired transactions after a second period of time, wherein there results a positive significant difference in a first price prediction and a negative significant difference in a second price prediction.","9.The method as claimed in claim 1, further comprising a step for determining the chance of a successful price prediction by relating a number of significant price predictions to the total number of price predictions.","10.The method as claimed in claim 1, further comprising a step for supplying one or more of said data to principals.","11.The method as claimed in claim 1, wherein the transactions are purchase and/or sale transactions.","12.The method as claimed in claim 8, wherein the second period of time is a period of for instance two weeks.","13.A system of one or more computers coupled via one or more networks to one or more other computers for performing the method as claimed in claim 1.14.A system comprising one or more computers, comprising means for performing a method for obtaining data relating to the sentiment of transactions on an (electronic) stock exchange, further comprising: input means for entering by a potential principal for stock exchange transactions electronically to one or more computers with one or more memories for storing therein one or more databases of transactions considered by the potential principal for a determined time period, and processing means for subsequently, i.e.","after a predetermined closing time for entering the transactions, determining a stock exchange sentiment on the basis of the content of the databases.","15.The system as claimed in claim 14, further comprising input means for performing a step for inputting of data by the principal concerning the actually performed transactions.","16.The system as claimed in claim 14, further comprising processing means for performing a step for determining differences between intended transactions and performed transactions.","17.The system as claimed in claim 14, further comprising input means for performing a step for inputting reasons for differences between intended transactions and performed transactions.","18.The system as claimed in claim 14, further comprising means comprising a parameter wherein the period of time is a day.","19.The system as claimed in claim 14, comprising: processing means for determining a fund score in respect of a stock exchange fund by relating data concerning desired transactions in a period of time to data concerning desired transactions in a subsequent period, wherein the closing price of the day is stored in the database if it is a significant sentiment.","20.The system as claimed in claim 18, further comprising processing means for determining that a sentiment is significant if there is a difference in fund score of at least a predetermined percentage.","21.The system as claimed in one or more of the claim 20, further comprising processing means for performing a step for determining a price prediction by relating the closing price from the databank in respect of desired transactions of the day to the closing price in respect of desired transactions after a second period of time, wherein there results a positive significant difference in a first price prediction and a negative significant difference in a second price prediction.","22.The system as claimed in claim 14, further comprising processing means for performing a step for determining the chance of a successful price prediction by relating a number of significant price predictions to the total number of price predictions.","23.The system as claimed in claim 14, further comprising processing means for performing a step for supplying one or more of said data to principals.","24.The system as claimed in claim 14, wherein the transactions are purchase and/or sale transactions.","25.The system as claimed in claim 21, wherein the second period of time is a period of for instance two weeks."],["Trading on the stock exchange has grown sharply in recent years and the share of private individuals herein has increased considerably and is still growing.","Partly owing to the development of the internet this will increase even further, since by making use of internet information which heretofore was only accessible to professionals also becomes accessible in timely manner to private individuals.","In addition, also due to internet, transaction costs have fallen considerably.","It is the expectation that these transaction costs will decrease still further.","Because of these developments private individuals are forming an increasingly important part of the stock exchange and are having more and more influence on price formation.","Research has shown that there is a large discrepancy between the evaluation of a fund by private individuals and professional analysts or traders.","It would therefore be very interesting to gain insight into the thinking of the private investor.","The present invention provides a method for obtaining data relating to the sentiment of transactions on an (electronic) stock exchange, comprising the following steps of: entering by a potential principal for stock exchange transactions electronically to one or more computers with one or more memories for storing therein one or more databases of transactions considered by this potential principal for a determined time period; and subsequently, i.e.","after a predetermined closing time for entering the transactions, determining a stock exchange sentiment on the basis of the content of the databases.","It hereby becomes the possible to automatically record which transactions an investor on the stock exchange is considering.","A great advantage hereof is that the attitude of a large number of investors in respect of determined funds is recorded on the basis of a large number of these considerations, on the basis of which stock exchange sentiments of this group can be determined.","This type of sentiment data can have, optionally after operations thereon, a predictive value, for instance for prices.","A preferred embodiment of the method further comprises a step of inputting by the principal of data concerning the actually performed transactions.","This embodiment has the advantage that particularly the intentions of the investor can be compared to the actual execution thereof.","If there is similarity between them, the intentions can for instance be assigned a higher predictive value.","If the intentions do not develop into actually performed transactions, the reasons herefor can be used to gain more insight into the relation between intentions and an execution thereof.","Another important advantage of having data concerning actually performed transactions is that analyses can be performed thereon.","A further preferred embodiment of the present method comprises a step for inputting reasons for differences between intended transactions and performed transactions.","It may for instance be important for a picture to be formed of the reasons for differences between the intentions of investors and the reasons that these intentions are not carried out or, on the contrary, are carried out.","A particular preferred embodiment provides that the period of time is a day.","It may for instance be useful to carry out daily measurements and to store the data of these measurements daily for the purpose of the method.","It is however very well possible to choose another period of time, depending on for instance the liveliness of the trading in a fund.","For the purpose of carrying out analyses, a particular embodiment of the method provides steps for: determining a fund score in respect of a stock exchange fund by relating data concerning desired transactions in the period of time to data concerning desired transactions in a subsequent period, wherein the closing price of the day is stored in the database if it is a significant sentiment.","An advantage hereof is that only data is stored relating to significant sentiments.","It is for instance much more useful to obtain information about rapid value increases or value decreases than about marginal developments.","A further preferred embodiment of the present invention provides a method which further comprises a step for determining a price prediction by relating the closing price from the databank with funds in respect of desired transactions of the first period to the closing price of funds in respect of desired transactions after a second period of time, wherein there results a positive significant difference in a first price prediction and a negative significant difference in a second price prediction.","An advantage hereof is that shorter term developments, such as the above described sentiments concerning the differences between two successive days, can be compared to longer term developments, such as for instance developments of the sentiment over a period of two weeks.","The first price prediction can for instance be given the value one and the second price prediction can for instance be given the value zero.","If a price prediction is significant, and thereby acquires the value one, this means that the sentiment is developing in positive sense in the case of a purchase sentiment, or that the sentiment is developing in negative sense in the case of a selling sentiment.","If a price prediction is not significant, and thereby acquires the value zero, this means that the sentiment is changing minimally, whereby the predictive value is low.","A further embodiment of the present invention is a method, which further provides a step for determining the chance of a successful price prediction by relating a number of significant price predictions to a total number of price predictions in a determined period.","This has the advantage that in respect of predictions a determination is made on the basis of real data from the past relating to how great is the reliability of predictions.","On the basis of the chance of a successful price prediction in a fund an estimate can be made as to the reliability of the sentiment data of the investors.","This is a powerful support means in taking decisions.","A further preferred embodiment according to the present invention provides a step for supplying one or more of said data to principals.","This can take place automatically by making use of computers on the basis of data in the databanks.","A further preferred embodiment of the present invention provides that the transactions are purchase and/or sale transactions.","This has the advantage that all the above stated steps can relate to both purchase and sale transactions.","A further preferred embodiment of the present method has the feature that the second period of time is a period of for instance two weeks.","This makes it possible to compare sentiment differences in a short term of for instance one day to developments in a longer term such as for instance the said two weeks.","It is certainly conceivable for both period lengths to be changed in order to obtain predictions which are preferred in specific situations.","A further preferred embodiment of the present method provides a system of one or more computers coupled via one or more networks to one or more other computers for performing the method.","Further advantages, features and details of the present invention will become apparent upon reading of the following description which refers to the drawings, wherein: FIG.","1 shows a block diagram of an example of a computer system for performing the method of the present invention; FIG.","2 is a block diagram showing a preferred embodiment of the present invention; FIG.","3 is a block diagram showing another preferred embodiment; FIG.","4 is a block diagram showing another preferred embodiment; FIG.","1 shows a computer system 1 that consists inter alia of a central processing unit 2 and a memory 3.This computer 1 is connected to the internet 4.The internet 4 is used to connect computer 1 to computers 5, 6 and 7.Computer 5 is used to input data about stock exchange transactions by potential principals.","These potential principals are people who effect purchase and sale transactions on the stock exchange, in particular private investors.","Computer 6 is preferably a computer of a supplier of price information.","Computer 6 of the stock exchange sends price information via the internet to computer 1.Computer 7 is the computer of parties other than the stock exchange and the investors who wish to receive information generated using the method.","These parties may for instance be the companies which have their shares listed on the stock exchange.","It may be interesting for these companies to obtain data concerning the sentiments of the investors about their share.","A reason for this may be that the investors influence the price by means of these sentiments.","An investor makes contact in FIG.","2 with the internet.","He logs in on a system of the method in 11 in order to be able to begin inputting data.","Another manner of inputting data is for instance via e-mail messages in 12.The investor or respondent then enters his share portfolio in 13, since an investor preferably only inputs an intention to sell in respect of a fund that he also possesses.","In 14 this data is stored in a database on computer 1.In 15 the investor indicates which funds he wishes to sell within a specific predetermined period.","This data is stored in a database in 16.In 17 the investor indicates which shares he wishes to purchase and in which quantities within a specific predetermined period.","In the case of purchase the investor can name all funds which are negotiable on the stock exchange and which are used in the system.","The data entered in 17 is stored in a databank in 18.The data inputted in 13, 15 and 17 is compared in 19 to data inputted during previous log-on sessions.","The changes in the mutations are further compared to changes in the portfolio in 20.If it is determined in 21 that the indicated changes have been implemented in the portfolio by means of transactions, these changes in the databases are processed in 22 as having been executed.","If it is found in 21 that the change has not been implemented in the portfolio, the investor is asked for the reason why.","This reason is added in 24 to the database.","FIG.","3 shows how information is sent to the computers 5, 6 and 7 by computer 1.Calculations are performed in 31 per stock exchange fund.","These calculations are preferably performed during off-peak periods, for instance at night, but can also be performed in real time during peak hours.","The calculations relate to the sentiments of the investors.","Examples of calculations which are performed: the percentage of the participants which possesses a fund relative to the total number of participants (number of possessors of the share divided by the total number of participants times 100%); unweighted sale sentiment is the percentage which is considering a sale relative to the number that possesses a relevant fund (number of persons who are thinking of selling divided by the number of persons possessing the share times 100%); unweighted purchase sentiment is the percentage which is thinking of purchasing relative to the total number of respondents (number of persons who are thinking of purchasing divided by the total number of respondents times 100%); weighted purchase sentiment is the number of shares which the participants are considering purchasing relative to a predefined measuring point (total number of shares which are possibly bought divided by predetermined figure times 100%); weighted sale sentiment is the total number of shares which the respondents are considering selling relative to a predefined measuring point (total number of shares which are possibly sold divided by predetermined figure times 100%); execution is the percentage of expected mutations which are carried out (number of mutations performed divided by the number mutations performed plus the number of mutations not performed for other reasons times 100%); a fund score for purchase, which means the number of potential purchases of today divided by the number of potential purchases of yesterday times 100%).","If this is significant (for instance 105%), the closing price of the dag later) the new closing price of this fund is recorded and the so-called price prediction calculation is performed thereon.","This price prediction calculation means that the price two weeks after the significant difference in the transaction score for the purchase divided by the price of the day that the fund score for the purchase differs significantly×100%.","If this price amounts to 105% or more at the moment of a purchase, it is deemed a good, significant prediction and this is designated in the database as a 1.If this price is less than 105% at the moment of a purchase, this is not a good, significant prediction and is designated in the database with a “0”.","In similar manner such fund scores are calculated for potential sales.","In this case the fund score is the result of the calculation of potential sales of today divided by potential sales of yesterday times 100%.","If this is significant (for instance 105%), the closing price of the day must be recorded.","A determined period (for instance two weeks) later, the new price of this fund is recorded and a calculation is also performed thereon.","This price prediction calculation proceeds in analogous manner as above: price of two weeks after significant difference in fund score divided by the price of the day that the fund score differs significantly times 100%.","If in the case of a sale fund score the price comes to less than for instance 95%, this is designated as good and the prediction is stored in the database with a value 1.If in the case of a sale fund score price calculation the result, the prediction, is more than 95%, this is designated as not good and the value 0 is stored in the database.","A chance of a successful prediction is defined as the number of times that a fund score differs significantly relative to the preceding period (for instance 105% of the previous day) and the number of times that a price also differs significantly (for instance 105%) after the period (of for instance two weeks).","This chance of a successful prediction is calculated by dividing the number of good predictions (1) by the total number of predictions (0 or 1) times 100%.","After performing the calculations per fund in 31, this data is stored in 32 in one or more of the databases with data of the time periods.","After construction the databases it is possible to distribute parts thereof to one or more of the computers 5, 6, 7 or other computers not participating in this system.","For instance in 34 data relating to a change in the sentiment is sent to such other computers.","Data is sent to computers 7 in 33.In 35 verification takes place as to which investors, or respondents, have logged in via computers 5.On the basis of data in the databases it is determined in 36 which investors are sent data concerning which funds.","There is no automatic right to all data, this depending on the agreements made in this respect.","It is determined in 37 which funds an investor has in portfolio.","It is for instance determined in 38 which possible purchases the investor has inputted, whereafter on the basis thereof data of the funds which result among others from the above described calculations are linked in 39 to data of the investor.","The investor can then receive this data in 40 by means of for instance an e-mail or a personalized web page.","FIG.","5 indicates a time span by means of 56, 51, 53 and 55.These blocks each represent moments.","In 50, which corresponds with day X, the investors input the above described data, such as portfolio-content and considered purchases and sales.","In 52, which corresponds in time with 53, being day X+1, the investor once again inputs data.","Feedback is also sent in 52 by the computer 1 which relates to the data of the day X in 51 and the data relating to the day X−1 of 56.These are the results of the calculations relating to differences between the preceding two consecutive days.","Calculations are also provided which relate to differences over a longer period, such as for instance two weeks.","In 54, which corresponds with day X+two weeks in 55, data is inputted by the investor.","If on the basis of this data the above described price has changed significantly, the chance of a successful prediction can be determined.","The results of this type of calculation will be sent a day later (not shown) to computers 5, 6, 7 or optional other computers.","Several statistical methods of analysis can be used for accessing whether e.g.","a chance of successful prediction or other indicators according to the invention achieve significant better or worse results than is to be expected.","It is useful to access the number of times a prediction has come true.","After entering a sentiment by a person several possibilities arise: 1.a purchase sentiment has been entered and the price has risen, 2.a purchase sentiment has been entered and the price is lowered, 3.a purchase sentiment is entered and the price has been remained equal, 4.a sell sentiment is entered and the price has risen, 5.a sell sentiment has been entered and the price has lowered, 6.a sell sentiment has been entered and the price has remained equal.","In case 1 and 5 the prediction has been right and in case en 2 and 4 the prediction has been incorrect.","Cases 3 en 6 in which the prices have remained equal are removed from de data.","What remains is a number of correct predictions and a number of incorrect predictions.","A change of a successful prediction is defined in formula 1: Percentage right = number of right predictions total number predictions * 100 % In principle the chance of a correct prediction is 0,5.Therefore it is to be expected that the number of correct predictions will be around 50%.","By calculating this percentage per fund it can be verified whether this is the case.","Prediction categories whereby this percentage deviates from the 50% point are interesting for further examination.","A first statistical test that is applied is the paired sample sign test.","What is being calculated is when the percentage of correct prediction is sufficiently far from 50% that the proposition that the sample is actually better or worse than the expected 50% can be upheld.","What is being tested is g: the number of rights predictions, whereby n=the number of values, p=the chance a prediction is correct.","The hypothesis is: H 0 : P g = 1 2 ( The chance of a correct prediction is equal to 1 2 ) H 1 : P g ≠ 1 2 ( The chance that a correct prediction is not equal to 1 2 ) If the null hypothesis is valid than g has a binomial distribution with n=the number of values and p = 1 2 .","Therefore the expectation of g equals: E ( g _ ) = 1 2 * n The standard deviation is: σ ( g _ ) = 1 2 * 1 2 * n Therefore the Z-score is: Z = g _ + 1 2 - E ( g _ ) σ ( g _ ) = g _ + 1 2 - ( 1 2 * n ) 1 2 * 1 2 * n Associated with this Z-score is a Z-value that is being calculated using the standard normal distribution P(z≧Z).","If this p-value is smaller than 0,05 than the null hypothesis is being dismissed.","A further way of assessing the predictions is by looking at the achieved return.","The return per prediction is being calculated as follows: Return per prediction = Price after evaluation period - Price at the time of entering the sentiment Price at the time of entering sentiment * 100 % With these returns the average return is calculated: Average return = Sum of returns per data element devided by the number of data elements Total number of data elements By evaluating either the right number of prediction or the return of the data elements right and wrong predictions can be compared to each other.","The expected average return is 0%.","By calculating the average return per prediction category it can be assessed whether this is the case.","The Wilcoxon signs rank test can be used for assessing whether a prediction category is significantly better or worse than the expected return of 0%.","The method is as follows: Determine for each data element the absolute return and remember whether this return has a plus sign or a minus sign.","Arrange the returns from low to high (without regarding the plus or minus signs).","Thus the lowest absolute deviation is assigned rank number 1 and the highest absolute deviation is assigned the highest rank number n. Add all the rank numbers of the deviations that originally had a plus sign and call this sum T+.","The sum of all the rank numbers that belong to deviations originally having a minus sign is T−.","If the null hypothesis is valid than it is to be expected that the T+ en T− are by and large equal.","If this is not the case than this is a signal that the return is significantly higher or lower than 0%.","The null hypothesis is: H0: μR=0 (The average return is 0%) The alternative hypothesis is: H1: μR≠0 (The average return is lower or higher than 0%) For assessing whether T+ assumes a value that is too high or too low the distribution of T+ has to be examined.","The sum of all rank numbers is: 1+2+ .",".",".","+n=0.5n(n+1) Therefore: T++T−=0.5n(n+1) If the null hypothesis with the test is valid than the chance of a positive and a negative deviation is equal to 0,5.Therefore the value that is to be expected for T+ is: E ( T + ) = 1 2 n ( n + 1 ) * 1 2 = n ( n + 1 ) 4 The standard deviation of T+ is: σ ( T + ) = n ( n + 1 ) ( 2 n + 1 ) 24 Herewith the Z-score can be calculated: Z = T + + 1 2 - E ( T + ) σ ( T + ) = T + + 1 2 - n ( n + 1 ) 4 n ( n + 1 ) ( 2 n + 1 ) 24 When the p-value that belongs to this Z-value is smaller than 0,05 than the null hypothesis is being rejected.","After assessing whether the properties of the prediction categories have significant better or worse results, a possible coherence between the prediction categories and the result is assessed.","What needs to be assessed is whether certain combinations of characteristics occur significantly more often than others.","For processing the coherence, e.g.","the Chi-square test is applied.","The Chi-square test is difficult to interpret but can serve as a basis for further ways of association.","In determining the Chi-square, firstly it is determined how the table would be if it were compiled entirely by chance, and no coherence would exist.","Thereafter, this table is compared with the empirical table and the degree of coherence is determined.","The Chi-square is calculated with the following formula: χ 2 = ∑ i = 1 R ∑ j = 1 C ( O ij - E ij ) 2 E ij in which: R=the total number of rows in the table, C=the total number of columns in the table, Oij=the observed value in row I, column j, Eij=the expected value in row i, column j. with E ij = f i .","* f .","j N in which: fi.=is the row total of row i, f.j=is the column total of column j, N=is the total number of samples.","The larger de Chi-square the bigger the coherence is between both variables.","Another statistical test thatlis applied is Cramérs V. Cramérs V is a major of association that is based on the Chi-square.","However, the number of samples and the size of the tables is being taken into account.","Therefore the value of Cramérs V is between 0 in absence of any coherence and 1 in case of full coherence.","Therefore the coherence between several variables can be compared.","Cramérs V is calculated using the formula: V = χ 2 N * min ( r - 1 , c - 1 ) in which: χ2=The Chi-square N=The total number of observations min(r-1,c-1)=the minimum of the number of rows or columns minus 1 Other methods for determining coherence between two variables are the Goodman and Kruskal's τ.","The Goodman and Kruskal's τ is a measure of an association that is based on the PRE-principle.","The letters PRE stand for the concept of “Proportional Reduction of Errors”.","PRE measures are a-symmetrical measures.","Therefore, it is necessary to designate a dependent variable beforehand.","In this case this would be the price after the evaluation period.","The formula for calculating the Goodman and Kruskal's τ is: τ = E 1 - E 2 E 2 in which: E 1 = ∑ i = 1 R f i .","* N - f i .","N E 2 = ∑ j = 1 C ∑ i = 1 R O ij * f .","j - O ij f .","j in which: R=The total number of rows in the table, C=The total number of columns in the table, Oij=The observed value in row i, column j, fi.=Row total of row i, f.j=Column total of column j, N=The total number of observations.","The value of the Goodman and Kruskal's τ is a percentage.","This percentage indicates the degree of influence of the independent variable on the dependent variable.","So if the Goodman and Kruskal's τ of variable X and variable Y is 5%, this means that adding knowledge about X will diminish the number of wrong predictions by 5%.","Further methods of analysis that can be applied include all prediction categories together, such as variance analysis or discriminant analysis.","Using variance analysis it is tested whether the expectation μ for a number of populations can have the same value.","It can be assessed whether the categories show significant differences within a dimension.","Therefore, it can be shown whether the dimension time influences the power of prediction.","Discriminant analysis is a variant of regression analysis, whereby the most notable difference has to do with the character of the dependent variable Y.","Using discriminant analysis a dependent variable with a nominal skill is being used.","In these methods all variables are also assessed at once.","An example of result that were achieved using the method are: Number Perc.","Average Time observations Right return 2 wk 233 0.714 0.058 4 wk 61 0.902 0.143 12 wk 97 0.711 0.117 In which the 233 entered sentiments where being entered by 38 different persons, the 61 entered sentiments were entered by 5 different persons, the 97 entered sentiments were entered by 13 different persons, and the total number of 391 entered sentiments where entered by 40 different persons.","The present invention is not limited to the described preferred embodiment; the rights sought are defined by the following claims."]],"string":"[\n [\n \"Method for obtaining data relating to the sentiment on a stock exchange\",\n \"The present invention relates to a method for obtaining data relating to the sentiment of transactions on an (electronic) stock exchange, comprising the following steps of:—entering by a potential principal for stock exchange transactions electronically to one or more computers with one or more memories for storing therein one or more databases of transactions considered by this potential principal for a determined time period; and subsequently, i.e.\",\n \"after a predetermined closing time for entering the transactions, determining a stock exchange sentiment on the basis of the content of the databases.\"\n ],\n [\n \"1.A method for obtaining data relating to the sentiment of transactions on an (electronic) stock exchange, comprising the steps of: entering by a potential principal for stock exchange transactions electronically to one or more memories for storing therein one or more databases of transactions considered by this potential principal for a determined time period; and subsequently, i.e.\",\n \"after a predetermined closing time for entering the transactions, determining a stock exchange sentiment on the basis of the content of the databases.\",\n \"2.The method as claimed in claim 1, further comprising a step of inputting by the principal of data concerning the actually performed transactions.\",\n \"3.The method as claimed in claim 1, further comprising a step for determining differences between intended transactions and performed transactions.\",\n \"4.The method as claimed in claim 1, further comprising a step for inputting reasons for differences between intended transactions and performed transactions.\",\n \"5.The method as claimed in claim 1, wherein the period of time is a day.\",\n \"6.The method as claimed in claim 1, comprising a step for: determining a fund score in respect of a stock exchange fund by relating data concerning desired transactions in a period of time to data concerning desired transactions in a subsequent period, wherein the closing price of the day is stored in the database if it is a significant sentiment.\",\n \"7.The method as claimed in claim 6, wherein a sentiment is significant if there is a difference in fund score of at least a predetermined percentage.\",\n \"8.The method as claimed in claim 1, further comprising a step for determining a price prediction by relating the closing price from the databank in respect of desired transactions of the day to the closing price in respect of desired transactions after a second period of time, wherein there results a positive significant difference in a first price prediction and a negative significant difference in a second price prediction.\",\n \"9.The method as claimed in claim 1, further comprising a step for determining the chance of a successful price prediction by relating a number of significant price predictions to the total number of price predictions.\",\n \"10.The method as claimed in claim 1, further comprising a step for supplying one or more of said data to principals.\",\n \"11.The method as claimed in claim 1, wherein the transactions are purchase and/or sale transactions.\",\n \"12.The method as claimed in claim 8, wherein the second period of time is a period of for instance two weeks.\",\n \"13.A system of one or more computers coupled via one or more networks to one or more other computers for performing the method as claimed in claim 1.14.A system comprising one or more computers, comprising means for performing a method for obtaining data relating to the sentiment of transactions on an (electronic) stock exchange, further comprising: input means for entering by a potential principal for stock exchange transactions electronically to one or more computers with one or more memories for storing therein one or more databases of transactions considered by the potential principal for a determined time period, and processing means for subsequently, i.e.\",\n \"after a predetermined closing time for entering the transactions, determining a stock exchange sentiment on the basis of the content of the databases.\",\n \"15.The system as claimed in claim 14, further comprising input means for performing a step for inputting of data by the principal concerning the actually performed transactions.\",\n \"16.The system as claimed in claim 14, further comprising processing means for performing a step for determining differences between intended transactions and performed transactions.\",\n \"17.The system as claimed in claim 14, further comprising input means for performing a step for inputting reasons for differences between intended transactions and performed transactions.\",\n \"18.The system as claimed in claim 14, further comprising means comprising a parameter wherein the period of time is a day.\",\n \"19.The system as claimed in claim 14, comprising: processing means for determining a fund score in respect of a stock exchange fund by relating data concerning desired transactions in a period of time to data concerning desired transactions in a subsequent period, wherein the closing price of the day is stored in the database if it is a significant sentiment.\",\n \"20.The system as claimed in claim 18, further comprising processing means for determining that a sentiment is significant if there is a difference in fund score of at least a predetermined percentage.\",\n \"21.The system as claimed in one or more of the claim 20, further comprising processing means for performing a step for determining a price prediction by relating the closing price from the databank in respect of desired transactions of the day to the closing price in respect of desired transactions after a second period of time, wherein there results a positive significant difference in a first price prediction and a negative significant difference in a second price prediction.\",\n \"22.The system as claimed in claim 14, further comprising processing means for performing a step for determining the chance of a successful price prediction by relating a number of significant price predictions to the total number of price predictions.\",\n \"23.The system as claimed in claim 14, further comprising processing means for performing a step for supplying one or more of said data to principals.\",\n \"24.The system as claimed in claim 14, wherein the transactions are purchase and/or sale transactions.\",\n \"25.The system as claimed in claim 21, wherein the second period of time is a period of for instance two weeks.\"\n ],\n [\n \"Trading on the stock exchange has grown sharply in recent years and the share of private individuals herein has increased considerably and is still growing.\",\n \"Partly owing to the development of the internet this will increase even further, since by making use of internet information which heretofore was only accessible to professionals also becomes accessible in timely manner to private individuals.\",\n \"In addition, also due to internet, transaction costs have fallen considerably.\",\n \"It is the expectation that these transaction costs will decrease still further.\",\n \"Because of these developments private individuals are forming an increasingly important part of the stock exchange and are having more and more influence on price formation.\",\n \"Research has shown that there is a large discrepancy between the evaluation of a fund by private individuals and professional analysts or traders.\",\n \"It would therefore be very interesting to gain insight into the thinking of the private investor.\",\n \"The present invention provides a method for obtaining data relating to the sentiment of transactions on an (electronic) stock exchange, comprising the following steps of: entering by a potential principal for stock exchange transactions electronically to one or more computers with one or more memories for storing therein one or more databases of transactions considered by this potential principal for a determined time period; and subsequently, i.e.\",\n \"after a predetermined closing time for entering the transactions, determining a stock exchange sentiment on the basis of the content of the databases.\",\n \"It hereby becomes the possible to automatically record which transactions an investor on the stock exchange is considering.\",\n \"A great advantage hereof is that the attitude of a large number of investors in respect of determined funds is recorded on the basis of a large number of these considerations, on the basis of which stock exchange sentiments of this group can be determined.\",\n \"This type of sentiment data can have, optionally after operations thereon, a predictive value, for instance for prices.\",\n \"A preferred embodiment of the method further comprises a step of inputting by the principal of data concerning the actually performed transactions.\",\n \"This embodiment has the advantage that particularly the intentions of the investor can be compared to the actual execution thereof.\",\n \"If there is similarity between them, the intentions can for instance be assigned a higher predictive value.\",\n \"If the intentions do not develop into actually performed transactions, the reasons herefor can be used to gain more insight into the relation between intentions and an execution thereof.\",\n \"Another important advantage of having data concerning actually performed transactions is that analyses can be performed thereon.\",\n \"A further preferred embodiment of the present method comprises a step for inputting reasons for differences between intended transactions and performed transactions.\",\n \"It may for instance be important for a picture to be formed of the reasons for differences between the intentions of investors and the reasons that these intentions are not carried out or, on the contrary, are carried out.\",\n \"A particular preferred embodiment provides that the period of time is a day.\",\n \"It may for instance be useful to carry out daily measurements and to store the data of these measurements daily for the purpose of the method.\",\n \"It is however very well possible to choose another period of time, depending on for instance the liveliness of the trading in a fund.\",\n \"For the purpose of carrying out analyses, a particular embodiment of the method provides steps for: determining a fund score in respect of a stock exchange fund by relating data concerning desired transactions in the period of time to data concerning desired transactions in a subsequent period, wherein the closing price of the day is stored in the database if it is a significant sentiment.\",\n \"An advantage hereof is that only data is stored relating to significant sentiments.\",\n \"It is for instance much more useful to obtain information about rapid value increases or value decreases than about marginal developments.\",\n \"A further preferred embodiment of the present invention provides a method which further comprises a step for determining a price prediction by relating the closing price from the databank with funds in respect of desired transactions of the first period to the closing price of funds in respect of desired transactions after a second period of time, wherein there results a positive significant difference in a first price prediction and a negative significant difference in a second price prediction.\",\n \"An advantage hereof is that shorter term developments, such as the above described sentiments concerning the differences between two successive days, can be compared to longer term developments, such as for instance developments of the sentiment over a period of two weeks.\",\n \"The first price prediction can for instance be given the value one and the second price prediction can for instance be given the value zero.\",\n \"If a price prediction is significant, and thereby acquires the value one, this means that the sentiment is developing in positive sense in the case of a purchase sentiment, or that the sentiment is developing in negative sense in the case of a selling sentiment.\",\n \"If a price prediction is not significant, and thereby acquires the value zero, this means that the sentiment is changing minimally, whereby the predictive value is low.\",\n \"A further embodiment of the present invention is a method, which further provides a step for determining the chance of a successful price prediction by relating a number of significant price predictions to a total number of price predictions in a determined period.\",\n \"This has the advantage that in respect of predictions a determination is made on the basis of real data from the past relating to how great is the reliability of predictions.\",\n \"On the basis of the chance of a successful price prediction in a fund an estimate can be made as to the reliability of the sentiment data of the investors.\",\n \"This is a powerful support means in taking decisions.\",\n \"A further preferred embodiment according to the present invention provides a step for supplying one or more of said data to principals.\",\n \"This can take place automatically by making use of computers on the basis of data in the databanks.\",\n \"A further preferred embodiment of the present invention provides that the transactions are purchase and/or sale transactions.\",\n \"This has the advantage that all the above stated steps can relate to both purchase and sale transactions.\",\n \"A further preferred embodiment of the present method has the feature that the second period of time is a period of for instance two weeks.\",\n \"This makes it possible to compare sentiment differences in a short term of for instance one day to developments in a longer term such as for instance the said two weeks.\",\n \"It is certainly conceivable for both period lengths to be changed in order to obtain predictions which are preferred in specific situations.\",\n \"A further preferred embodiment of the present method provides a system of one or more computers coupled via one or more networks to one or more other computers for performing the method.\",\n \"Further advantages, features and details of the present invention will become apparent upon reading of the following description which refers to the drawings, wherein: FIG.\",\n \"1 shows a block diagram of an example of a computer system for performing the method of the present invention; FIG.\",\n \"2 is a block diagram showing a preferred embodiment of the present invention; FIG.\",\n \"3 is a block diagram showing another preferred embodiment; FIG.\",\n \"4 is a block diagram showing another preferred embodiment; FIG.\",\n \"1 shows a computer system 1 that consists inter alia of a central processing unit 2 and a memory 3.This computer 1 is connected to the internet 4.The internet 4 is used to connect computer 1 to computers 5, 6 and 7.Computer 5 is used to input data about stock exchange transactions by potential principals.\",\n \"These potential principals are people who effect purchase and sale transactions on the stock exchange, in particular private investors.\",\n \"Computer 6 is preferably a computer of a supplier of price information.\",\n \"Computer 6 of the stock exchange sends price information via the internet to computer 1.Computer 7 is the computer of parties other than the stock exchange and the investors who wish to receive information generated using the method.\",\n \"These parties may for instance be the companies which have their shares listed on the stock exchange.\",\n \"It may be interesting for these companies to obtain data concerning the sentiments of the investors about their share.\",\n \"A reason for this may be that the investors influence the price by means of these sentiments.\",\n \"An investor makes contact in FIG.\",\n \"2 with the internet.\",\n \"He logs in on a system of the method in 11 in order to be able to begin inputting data.\",\n \"Another manner of inputting data is for instance via e-mail messages in 12.The investor or respondent then enters his share portfolio in 13, since an investor preferably only inputs an intention to sell in respect of a fund that he also possesses.\",\n \"In 14 this data is stored in a database on computer 1.In 15 the investor indicates which funds he wishes to sell within a specific predetermined period.\",\n \"This data is stored in a database in 16.In 17 the investor indicates which shares he wishes to purchase and in which quantities within a specific predetermined period.\",\n \"In the case of purchase the investor can name all funds which are negotiable on the stock exchange and which are used in the system.\",\n \"The data entered in 17 is stored in a databank in 18.The data inputted in 13, 15 and 17 is compared in 19 to data inputted during previous log-on sessions.\",\n \"The changes in the mutations are further compared to changes in the portfolio in 20.If it is determined in 21 that the indicated changes have been implemented in the portfolio by means of transactions, these changes in the databases are processed in 22 as having been executed.\",\n \"If it is found in 21 that the change has not been implemented in the portfolio, the investor is asked for the reason why.\",\n \"This reason is added in 24 to the database.\",\n \"FIG.\",\n \"3 shows how information is sent to the computers 5, 6 and 7 by computer 1.Calculations are performed in 31 per stock exchange fund.\",\n \"These calculations are preferably performed during off-peak periods, for instance at night, but can also be performed in real time during peak hours.\",\n \"The calculations relate to the sentiments of the investors.\",\n \"Examples of calculations which are performed: the percentage of the participants which possesses a fund relative to the total number of participants (number of possessors of the share divided by the total number of participants times 100%); unweighted sale sentiment is the percentage which is considering a sale relative to the number that possesses a relevant fund (number of persons who are thinking of selling divided by the number of persons possessing the share times 100%); unweighted purchase sentiment is the percentage which is thinking of purchasing relative to the total number of respondents (number of persons who are thinking of purchasing divided by the total number of respondents times 100%); weighted purchase sentiment is the number of shares which the participants are considering purchasing relative to a predefined measuring point (total number of shares which are possibly bought divided by predetermined figure times 100%); weighted sale sentiment is the total number of shares which the respondents are considering selling relative to a predefined measuring point (total number of shares which are possibly sold divided by predetermined figure times 100%); execution is the percentage of expected mutations which are carried out (number of mutations performed divided by the number mutations performed plus the number of mutations not performed for other reasons times 100%); a fund score for purchase, which means the number of potential purchases of today divided by the number of potential purchases of yesterday times 100%).\",\n \"If this is significant (for instance 105%), the closing price of the dag later) the new closing price of this fund is recorded and the so-called price prediction calculation is performed thereon.\",\n \"This price prediction calculation means that the price two weeks after the significant difference in the transaction score for the purchase divided by the price of the day that the fund score for the purchase differs significantly×100%.\",\n \"If this price amounts to 105% or more at the moment of a purchase, it is deemed a good, significant prediction and this is designated in the database as a 1.If this price is less than 105% at the moment of a purchase, this is not a good, significant prediction and is designated in the database with a “0”.\",\n \"In similar manner such fund scores are calculated for potential sales.\",\n \"In this case the fund score is the result of the calculation of potential sales of today divided by potential sales of yesterday times 100%.\",\n \"If this is significant (for instance 105%), the closing price of the day must be recorded.\",\n \"A determined period (for instance two weeks) later, the new price of this fund is recorded and a calculation is also performed thereon.\",\n \"This price prediction calculation proceeds in analogous manner as above: price of two weeks after significant difference in fund score divided by the price of the day that the fund score differs significantly times 100%.\",\n \"If in the case of a sale fund score the price comes to less than for instance 95%, this is designated as good and the prediction is stored in the database with a value 1.If in the case of a sale fund score price calculation the result, the prediction, is more than 95%, this is designated as not good and the value 0 is stored in the database.\",\n \"A chance of a successful prediction is defined as the number of times that a fund score differs significantly relative to the preceding period (for instance 105% of the previous day) and the number of times that a price also differs significantly (for instance 105%) after the period (of for instance two weeks).\",\n \"This chance of a successful prediction is calculated by dividing the number of good predictions (1) by the total number of predictions (0 or 1) times 100%.\",\n \"After performing the calculations per fund in 31, this data is stored in 32 in one or more of the databases with data of the time periods.\",\n \"After construction the databases it is possible to distribute parts thereof to one or more of the computers 5, 6, 7 or other computers not participating in this system.\",\n \"For instance in 34 data relating to a change in the sentiment is sent to such other computers.\",\n \"Data is sent to computers 7 in 33.In 35 verification takes place as to which investors, or respondents, have logged in via computers 5.On the basis of data in the databases it is determined in 36 which investors are sent data concerning which funds.\",\n \"There is no automatic right to all data, this depending on the agreements made in this respect.\",\n \"It is determined in 37 which funds an investor has in portfolio.\",\n \"It is for instance determined in 38 which possible purchases the investor has inputted, whereafter on the basis thereof data of the funds which result among others from the above described calculations are linked in 39 to data of the investor.\",\n \"The investor can then receive this data in 40 by means of for instance an e-mail or a personalized web page.\",\n \"FIG.\",\n \"5 indicates a time span by means of 56, 51, 53 and 55.These blocks each represent moments.\",\n \"In 50, which corresponds with day X, the investors input the above described data, such as portfolio-content and considered purchases and sales.\",\n \"In 52, which corresponds in time with 53, being day X+1, the investor once again inputs data.\",\n \"Feedback is also sent in 52 by the computer 1 which relates to the data of the day X in 51 and the data relating to the day X−1 of 56.These are the results of the calculations relating to differences between the preceding two consecutive days.\",\n \"Calculations are also provided which relate to differences over a longer period, such as for instance two weeks.\",\n \"In 54, which corresponds with day X+two weeks in 55, data is inputted by the investor.\",\n \"If on the basis of this data the above described price has changed significantly, the chance of a successful prediction can be determined.\",\n \"The results of this type of calculation will be sent a day later (not shown) to computers 5, 6, 7 or optional other computers.\",\n \"Several statistical methods of analysis can be used for accessing whether e.g.\",\n \"a chance of successful prediction or other indicators according to the invention achieve significant better or worse results than is to be expected.\",\n \"It is useful to access the number of times a prediction has come true.\",\n \"After entering a sentiment by a person several possibilities arise: 1.a purchase sentiment has been entered and the price has risen, 2.a purchase sentiment has been entered and the price is lowered, 3.a purchase sentiment is entered and the price has been remained equal, 4.a sell sentiment is entered and the price has risen, 5.a sell sentiment has been entered and the price has lowered, 6.a sell sentiment has been entered and the price has remained equal.\",\n \"In case 1 and 5 the prediction has been right and in case en 2 and 4 the prediction has been incorrect.\",\n \"Cases 3 en 6 in which the prices have remained equal are removed from de data.\",\n \"What remains is a number of correct predictions and a number of incorrect predictions.\",\n \"A change of a successful prediction is defined in formula 1: Percentage right = number of right predictions total number predictions * 100 % In principle the chance of a correct prediction is 0,5.Therefore it is to be expected that the number of correct predictions will be around 50%.\",\n \"By calculating this percentage per fund it can be verified whether this is the case.\",\n \"Prediction categories whereby this percentage deviates from the 50% point are interesting for further examination.\",\n \"A first statistical test that is applied is the paired sample sign test.\",\n \"What is being calculated is when the percentage of correct prediction is sufficiently far from 50% that the proposition that the sample is actually better or worse than the expected 50% can be upheld.\",\n \"What is being tested is g: the number of rights predictions, whereby n=the number of values, p=the chance a prediction is correct.\",\n \"The hypothesis is: H 0 : P g = 1 2 ( The chance of a correct prediction is equal to 1 2 ) H 1 : P g ≠ 1 2 ( The chance that a correct prediction is not equal to 1 2 ) If the null hypothesis is valid than g has a binomial distribution with n=the number of values and p = 1 2 .\",\n \"Therefore the expectation of g equals: E ( g _ ) = 1 2 * n The standard deviation is: σ ( g _ ) = 1 2 * 1 2 * n Therefore the Z-score is: Z = g _ + 1 2 - E ( g _ ) σ ( g _ ) = g _ + 1 2 - ( 1 2 * n ) 1 2 * 1 2 * n Associated with this Z-score is a Z-value that is being calculated using the standard normal distribution P(z≧Z).\",\n \"If this p-value is smaller than 0,05 than the null hypothesis is being dismissed.\",\n \"A further way of assessing the predictions is by looking at the achieved return.\",\n \"The return per prediction is being calculated as follows: Return per prediction = Price after evaluation period - Price at the time of entering the sentiment Price at the time of entering sentiment * 100 % With these returns the average return is calculated: Average return = Sum of returns per data element devided by the number of data elements Total number of data elements By evaluating either the right number of prediction or the return of the data elements right and wrong predictions can be compared to each other.\",\n \"The expected average return is 0%.\",\n \"By calculating the average return per prediction category it can be assessed whether this is the case.\",\n \"The Wilcoxon signs rank test can be used for assessing whether a prediction category is significantly better or worse than the expected return of 0%.\",\n \"The method is as follows: Determine for each data element the absolute return and remember whether this return has a plus sign or a minus sign.\",\n \"Arrange the returns from low to high (without regarding the plus or minus signs).\",\n \"Thus the lowest absolute deviation is assigned rank number 1 and the highest absolute deviation is assigned the highest rank number n. Add all the rank numbers of the deviations that originally had a plus sign and call this sum T+.\",\n \"The sum of all the rank numbers that belong to deviations originally having a minus sign is T−.\",\n \"If the null hypothesis is valid than it is to be expected that the T+ en T− are by and large equal.\",\n \"If this is not the case than this is a signal that the return is significantly higher or lower than 0%.\",\n \"The null hypothesis is: H0: μR=0 (The average return is 0%) The alternative hypothesis is: H1: μR≠0 (The average return is lower or higher than 0%) For assessing whether T+ assumes a value that is too high or too low the distribution of T+ has to be examined.\",\n \"The sum of all rank numbers is: 1+2+ .\",\n \".\",\n \".\",\n \"+n=0.5n(n+1) Therefore: T++T−=0.5n(n+1) If the null hypothesis with the test is valid than the chance of a positive and a negative deviation is equal to 0,5.Therefore the value that is to be expected for T+ is: E ( T + ) = 1 2 n ( n + 1 ) * 1 2 = n ( n + 1 ) 4 The standard deviation of T+ is: σ ( T + ) = n ( n + 1 ) ( 2 n + 1 ) 24 Herewith the Z-score can be calculated: Z = T + + 1 2 - E ( T + ) σ ( T + ) = T + + 1 2 - n ( n + 1 ) 4 n ( n + 1 ) ( 2 n + 1 ) 24 When the p-value that belongs to this Z-value is smaller than 0,05 than the null hypothesis is being rejected.\",\n \"After assessing whether the properties of the prediction categories have significant better or worse results, a possible coherence between the prediction categories and the result is assessed.\",\n \"What needs to be assessed is whether certain combinations of characteristics occur significantly more often than others.\",\n \"For processing the coherence, e.g.\",\n \"the Chi-square test is applied.\",\n \"The Chi-square test is difficult to interpret but can serve as a basis for further ways of association.\",\n \"In determining the Chi-square, firstly it is determined how the table would be if it were compiled entirely by chance, and no coherence would exist.\",\n \"Thereafter, this table is compared with the empirical table and the degree of coherence is determined.\",\n \"The Chi-square is calculated with the following formula: χ 2 = ∑ i = 1 R ∑ j = 1 C ( O ij - E ij ) 2 E ij in which: R=the total number of rows in the table, C=the total number of columns in the table, Oij=the observed value in row I, column j, Eij=the expected value in row i, column j. with E ij = f i .\",\n \"* f .\",\n \"j N in which: fi.=is the row total of row i, f.j=is the column total of column j, N=is the total number of samples.\",\n \"The larger de Chi-square the bigger the coherence is between both variables.\",\n \"Another statistical test thatlis applied is Cramérs V. Cramérs V is a major of association that is based on the Chi-square.\",\n \"However, the number of samples and the size of the tables is being taken into account.\",\n \"Therefore the value of Cramérs V is between 0 in absence of any coherence and 1 in case of full coherence.\",\n \"Therefore the coherence between several variables can be compared.\",\n \"Cramérs V is calculated using the formula: V = χ 2 N * min ( r - 1 , c - 1 ) in which: χ2=The Chi-square N=The total number of observations min(r-1,c-1)=the minimum of the number of rows or columns minus 1 Other methods for determining coherence between two variables are the Goodman and Kruskal's τ.\",\n \"The Goodman and Kruskal's τ is a measure of an association that is based on the PRE-principle.\",\n \"The letters PRE stand for the concept of “Proportional Reduction of Errors”.\",\n \"PRE measures are a-symmetrical measures.\",\n \"Therefore, it is necessary to designate a dependent variable beforehand.\",\n \"In this case this would be the price after the evaluation period.\",\n \"The formula for calculating the Goodman and Kruskal's τ is: τ = E 1 - E 2 E 2 in which: E 1 = ∑ i = 1 R f i .\",\n \"* N - f i .\",\n \"N E 2 = ∑ j = 1 C ∑ i = 1 R O ij * f .\",\n \"j - O ij f .\",\n \"j in which: R=The total number of rows in the table, C=The total number of columns in the table, Oij=The observed value in row i, column j, fi.=Row total of row i, f.j=Column total of column j, N=The total number of observations.\",\n \"The value of the Goodman and Kruskal's τ is a percentage.\",\n \"This percentage indicates the degree of influence of the independent variable on the dependent variable.\",\n \"So if the Goodman and Kruskal's τ of variable X and variable Y is 5%, this means that adding knowledge about X will diminish the number of wrong predictions by 5%.\",\n \"Further methods of analysis that can be applied include all prediction categories together, such as variance analysis or discriminant analysis.\",\n \"Using variance analysis it is tested whether the expectation μ for a number of populations can have the same value.\",\n \"It can be assessed whether the categories show significant differences within a dimension.\",\n \"Therefore, it can be shown whether the dimension time influences the power of prediction.\",\n \"Discriminant analysis is a variant of regression analysis, whereby the most notable difference has to do with the character of the dependent variable Y.\",\n \"Using discriminant analysis a dependent variable with a nominal skill is being used.\",\n \"In these methods all variables are also assessed at once.\",\n \"An example of result that were achieved using the method are: Number Perc.\",\n \"Average Time observations Right return 2 wk 233 0.714 0.058 4 wk 61 0.902 0.143 12 wk 97 0.711 0.117 In which the 233 entered sentiments where being entered by 38 different persons, the 61 entered sentiments were entered by 5 different persons, the 97 entered sentiments were entered by 13 different persons, and the total number of 391 entered sentiments where entered by 40 different persons.\",\n \"The present invention is not limited to the described preferred embodiment; the rights sought are defined by the following claims.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10474262"}}},{"rowIdx":791,"cells":{"text":{"kind":"list like","value":[["Specific detection of proteolytic enzymes","Specific detection of proteolytic enzymes is achieved by extinguishing dye fluorescence by the amino acid tryptophan.","Tryptophan is disposed on one side of the cutting site of a proteolytic enzyme while an amino acid marked with a dye is arranged on the other side.","Extinction of fluorescence occurs prior to enzyme cutting.","Spatial separation of the tryptophan and the dye takes place after cutting, whereby fluorescence extinction does not occur.","The dye can then fluoresce and a signal increase occurs thereby indicating that cutting has been carried out and the presence of the enzyme."],["1.","(canceled) 2.The peptide as claimed in claim 21, wherein the fluorescent dye is one of an oxazine derivative or a rhodamine derivative.","3.A method for the detection of an endopeptidase comprising the following steps: mixing the peptide as claimed in claim 21 and an enzyme to be detected in a solution; and determining an intensity of emission of the fluorophore in the solution.","4.A device on which a peptide as claimed in claim 21 is immobilized.","5.A device for immobilizing the peptide as claimed in claim 21, wherein the peptide is immobilized on the device in such a way that after cleavage by the enzyme to be detected, the fluorophore remains immobilized on a surface, whereas the quencher is no longer immobilized on the surface.","6.","(canceled) 7.The peptide as claimed in claim 22, wherein the fluorescent dye is an oxazine derivative or a rhodamine derivative.","8.The peptide as claimed in claim 22, wherein: the quencher is at least one first fluorescent dye; the fluorophore is at least one second fluorescent dye; the at least one first and one second fluorescent dye can couple covalently to peptides or proteins; and the at least one first and second fluorescent dye can quench each other.","9.A method for the detection of an exopeptidase comprising the following steps: mixing a peptide as claimed in claim 22 and the enzyme to be detected in a solution; and determining the intensity of emission of the fluorophore in the solution.","10.A device on which a peptide as claimed in claim 22 s immobilized.","11.A device for immobilizing the peptide as claimed in claim 22, wherein the peptide is immobilized on the device in such a way that after cleavage by the enzyme to be detected, the fluorophore remains immobilized on the surface, whereas the quencher is no longer immobilized on the surface.","12.","(canceled) 13.","(canceled) 14.The peptide as claimed in claim 24, wherein the fluorescent dye is an oxazine derivative or a rhodamine derivative.","15.The peptide as claimed in claim 23, wherein the quencher is at least one first fluorescent dye; the fluorophore is at last one second fluorescent dye; the at least one first and second fluorescent dye can couple covalently to peptides or proteins; and the at least one first and second fluorescent dye can quench each other, 16.","(canceled) 17.A method for the detection of an endopeptidase comprising the following steps; mixing a peptide as claimed in claim 23 at least one exopeptidase and the endopeptidase to be detected, in a solution; and determining the intensity of emission of the fluorophore in the solution.","18.","(canceled) 19.","(canceled) 20.Kit comprising the peptide as claimed in claim 23 and including at least one exopeptidase.","21.A peptide with the following properties, the peptide comprising an identification sequence of an endopeptidase and functioning as substrate for the endopeptidase and providing a cleavage site between two amino acids at the identification sequence in the peptide; and at least one fluorophore in the form of a fluorescent dye for covalently coupling to peptides or proteins and located at one side of the cleavage site; and at least one quencher in the form of tryptophan arranged on the other side of the cleavage site; and wherein the quencher is sufficiently proximate to the fluorophore so that before the substrate at the cleavage site between quencher and fluorophore is cleaved, at least a partial quenching of a fluorescent emission from the fluorophore is realized; and wherein after the substrate is cleaved by the endopeptidase at the cleavage site, emission by the fluorophore is increased.","22.A peptide with the following properties, comprising: an amino acid sequence, and at least one group containing tryptophan as a quencher and a fluorescent dye as a fluorophore on different amino acids; wherein the fluorescent dye is able to couple covalently to peptides or proteins and to emit fluorescence that is quenched by tryptophan; wherein the sequence contains sufficient spatial proximity between the quencher and the fluorophore so that as long as the amino acids carrying the quencher and the fluorescent dye have not been separated by an exopeptidase, at least a partial quenching of the emitted fluorescence of the fluorophore is realized; and wherein after the amino acids carrying the quencher and the fluorescent dye are separated by the exopeptidase, at least a partial increase in fluorescent emission of the fluorophore is realized.","23.A peptide with the following properties, the peptide comprising: an identification sequence of an endopeptidase and functioning as substrate for the endopeptidase and providing a cleavage site between two amino acids at the identification sequence in the peptide; and a stop group that prevents an exopeptidase from digesting the peptide and arranged at one end of the peptide; and wherein the identification sequence includes at least one group containing a quencher and a fluorophore on different amino acids and arranged downstream from the stop group; wherein the quencher and the fluorophore are sufficiently proximate, so that as long as the amino acids carrying the quencher and the fluorescent dye have not yet been separated by an exopeptidase, at least a partial quenching of the emission of the fluorophore is realized; and wherein after the amino acids carrying the quencher and the fluorescent dye are separated by an exopeptidase, a spatial distance is realized between the quencher or any other quencher and the fluorophore sufficient to realize an at least partial increase in emission the fluorophore.","24.The peptide as claimed in claim 23, wherein the fluorophore is a fluorescent dye for coupling covalently to peptides or proteins and the quencher is tryptophan for quenching the fluorescence emitted from the fluorescent dye."],["Proteolytic enzymes are proteases or peptidases, i.e.","enzymes that are able to split or lyse peptides or proteins.","The word “peptide” generally denotes a shorter amino acid sequence, whereas the word “protein” generally denotes larger molecules consisting of an amino acid sequence.","Both terms are used synonymously in the following.","Peptidases and proteases play a decisive role in protein activation, cell regulation and signal transmission.","Their detection at maximum possible sensitivity is therefore of enormous importance for understanding the functioning of a cell and its communication, and for monitoring the course of diseases.","Furthermore, precise knowledge of peptidase or protease concentration permits the targeted development of new therapeutic agents for inhibition, e.g.","of HIV protease.","HIV protease breaks down the HIV protein, which is at first very long after multiplication in a cell, into functioning individual proteins, which only then permit the further activity and growth of the HIV virus.","There are two possible approaches.","Firstly, if the HIV protease is blocked, the HIV virus cannot replicate and its propagation is halted.","Secondly, infection with the HIV virus can take place via detection of the HIV protease.","Specific detection of HIV protease can therefore serve as an AIDS test.","Moreover, protease tests are attracting increasing attention in medical research, because more and more diseases, including cancers, are being connected with enzymes.","Tumor-associated proteases are increasingly the object of oncological-biochemical research.","These proteases bring about the breakdown of proteins of the tumor stroma and of the basal membrane and thus permit tumor cell invasion.","Therefore investigations of protease systems that are overexpressed in tumor cells are among the new prognosis factors in tumor diagnostics.","Another important aspect is the use of protease tests in quality control of biochemical products.","If, for example proteins, enzymes, peptides, etc.","are being sold, it is necessary to ensure that the products sold are free from proteases, so that the product sold does not break down on its own.","The same can be used for testing the specific activity of proteases, e.g.","after prolonged storage, regarding the question as to whether the enzymes are still active.","The proteolytic enzymes are divided into endopeptidases (proteases) and exopeptidases.","Endopeptidases cleave amino linkages inside peptides.","Exopeptidases digest peptides amino acid by amino acid starting from their ends, i.e.","they cleave terminal amino acids.","The exopeptidases can be further subdivided into aminopeptidases and carboxypeptidases, as claimed in their activity at the amino or N end or at the carboxy or C end of the peptide.","For determination of the concentration or activity of various peptidases and proteases, to date various fluorescence-based tests have been developed, and their operating principles and sensitivities are briefly described below.","One possibility for detecting proteolytic enzymes is offered by the “EnzChek Protease Assay Kit” from the company Molecular Probes (Eugene, USA).","For detecting proteases, this kit employs casein derivatives, to which very many pH-insensitive green or red fluorescing BODIPY dyes are coupled (Karolin J., Johansson B.","A., Strandberg L., Ny T.; J.","Am.","Chem.","Soc., 1994, 116, 7801-7806).","On account of the very high degree of marking of the protein, the intermolecular distances between the fluorophores are very small (typically of the order of a few nanometers).","The dyes therefore extinguish each other, inter alia by dimer formation.","Casein is a large enzyme, which contains many binding sites for proteases, because it has many different sequence segments.","If the dye-marked casein derivative comes into contact with proteases, e.g.","with trypsin, which hydrolyzes peptide bonds of the basic amino acids arginine and lysine on the carboxy side, these split the casein derivative into smaller peptides, so that as a rule the distance between the dyes increases.","Extinction does not occur, and an increase in fluorescence is observed, indicating that proteolytic enzymes are present.","For trypsin, for example, the detection sensitivity of this kit is a few ng/ml.","For these small quantities of protease the detection time or measurement time is more than 10 hours.","This test detects many proteolytic enzymes nonspecifically, since casein is digested by elastase, pepsin, thermolysin, papain, trypsin, etc.","An enzyme test based on Rhodamine 110 for various proteases and peptidases (Leytus S. P., Melhado L. L., Mangel W. F.: Rhodamine-based compounds as fluorogenic substrates for serine protease, (1983) Biochem.","J., 209 (2): 299-307; Leytus S. P., Patterson W. L., Mangel W. F.: New class of sensitive and selective fluorogenic substrates for serine proteinases.","Amino acid and dipeptide derivatives of rhodamine, (1983) Biochem.","J., 215 (2): 253-260) is also being marketed for the detection of serine proteases.","In this test, at least one of the amino groups of Rhodamine 110 is coupled covalently to an amino acid (usually arginine), causing strong extinction of the fluorescence of the fluorophore.","After digestion of the peptide bonds there is a dramatic increase in fluorescence of Rhodamine 110.For caspase-3, the sensitivity of the test is a few ng absolute quantity of enzyme.","Disadvantages are that the principle is not of general applicability, sometimes its action is nonspecific, and it is relatively insensitive.","A protease or peptidase test can also be accomplished by means of Forster fluorescence resonance energy transfer (FRET) (Forster Th.",": Zwischenmolekulare Energiewanderung und Fluoreszenz [Intermolecular Energy Transfer and Fluorescence], (1948) Annalen der Physik, 2: 55-75).","In this, the specific identification sequence of an endopeptidase is marked covalently at both ends with a donor dye and an acceptor dye.","Owing to spectral overlap of donor emission with acceptor absorption, the fluorescence of the donor is extinguished at small distances.","If the peptide is cleaved by the endopeptidase, the spatially close contact between donor and acceptor is lost, leading to an increase in fluorescence.","Detection of HIV protease will be briefly explained here, as an example.","The specific cleaving sequence of HIV protease is coupled, near the cleavage site, with an EDANS dye (donor) and a DABCYL dye (acceptor).","The peptide sequence is then: Arg-Glu(EDANS)-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys-(DABCYL)-Arg, using the 3-letter abbreviations for the amino acids.","After cleavage by the HIV protease we have two parts: Arg-Glu(EDANS)Ser-Gln-Asn-Tyr-OH and Pro-Ile-Val-Gln-Lys(DABCYL)-Arg.","The principle is of general applicability, but there must be a suitable small distance between the two dyes.","Sensitivity is of the order of nanomolar solutions of HIV protease.","A disadvantage is the expensive synthesis of the substrate with specific coupling of a donor and an acceptor.","If the substrate is to some extent marked incompletely, e.g.","with just one donor, this leads to poor sensitivity of the test.","The aim of the invention is to improve the possibilities for a specific detection of proteolytic enzymes.","This aim is achieved by the inventions as claimed in the independent claims.","Advantageous developments are described in the subclaims.","A first method of detection (“test” for short) is used for the detection of endopeptidases (also briefly referred to as enzymes hereinafter) by means of special peptides.","For this purpose, a peptide is prepared with the properties enumerated below.","The peptide contains an identification sequence of the endopeptidase that is to be detected, and can therefore serve as substrate for the enzyme, with a cleavage site between two amino acids in the identification sequence in the peptide.","The peptide contains at least one quencher and at least one fluorophore.","“Quenching” means reduction of the emission of the fluorophore due to interaction with the aforesaid quencher.","A fluorescent dye is used as the fluorophore.","Quenching causes a decrease in fluorescence quantum yield.","The words “extinction” and “extinguishing molecule” or “extinguisher” are also used as synonyms for “quenching” and “quencher”.","The amino acid tryptophan is used as quencher.","Other amino acids, such as tyrosine or histidine, also exhibit slight fluorescence extinction, but do not have the strong quenching action of tryptophan.","The fluorescent dye is selected on the basis that its fluorescence can be extinguished by tryptophan.","Furthermore, it should be possible for the fluorescent dye to be coupled covalently to peptides or proteins.","As a rule the fluorophore is coupled to an amino acid.","Attainment of high sensitivity of the test requires targeted selection of the peptide sequence: the quencher must be arranged on one side of the cleavage site, with the fluorophore on the other side of the cleavage site.","The sequence must be selected so that the quencher and the fluorophore are sufficiently spatially close to one another, so long as the substrate has still not been cleaved by the enzyme at the cleavage site between quencher and fluorophore, to provide the possibility of at least partial extinction of the emission of the fluorophore.","After cleavage of the substrate by the enzyme at the cleavage site between quencher and fluorophore, a definite increase in emission of the fluorophore must be possible.","Thus, the peptide or protein is not marked with the fluorescent dye on the tryptophan itself, but on another amino acid, which is separated from the tryptophan by the cleavage site.","Furthermore, an amino acid other than tryptophan is chosen for coupling the fluorescent dye, e.g.","lysine, arginine or cysteine.","The fluorescent dye can also be coupled to one end of the peptide or protein.","The largest increase in fluorescence is obtained if the tryptophan and the dye-marked amino acid are located directly on either side of the cleavage site.","For dye MR 121, for example, an increase in fluorescence by a factor of 10 can be observed after cleavage of the peptide by the enzyme.","Moreover, no tryptophan is to be arranged in adjacent positions on the same side of the cleavage site of the peptide sequence as the dye, and especially not in immediately adjacent positions, so that the dye is not quenched by this additional tryptophan.","Appropriate choice of substrate sequence makes it possible to mark the substrate with just one fluorescent dye, and not with at least two, as was necessary until now.","This simplifies the synthesis of the substrate.","In addition there is less disturbance of binding of the enzyme to the substrate by coupled dyes, resulting in greater specificity of binding between substrate and enzyme.","Finally, the peptide thus prepared and the enzyme to be detected are mixed in a solution.","If binding occurs between peptide and peptidase, the peptide is cleaved at the cleavage site.","As a result, the spatial distance between the fluorophore and the quencher becomes greater, and the emission of the fluorophore increases.","This increase in emission is determined and serves for detecting those peptidases or proteases that bind to the identification sequence and can cleave the peptide there.","Accordingly, the detection reaction is specific.","It can be directed towards the detection of specific enzymes by varying the peptide sequence.","In particular, the test can detect enzymes specifically, even when other types of enzymes are also present in the solution.","For example, the enzyme trypsin can be detected in a solution that also contains elastase and chymotrypsin.","If the temporal course of the increase in emission is traced, the concentration of the enzyme can be determined quantitatively.","Using the test as claimed in the invention, it is even possible to detect proteases or peptidases at a concentration of 10{circumflex over ( )}−15 mol/l.","As dyes, consideration may be given to all fluorescent dyes that are measurably extinguished in their fluorescence by the amino acid tryptophan.","Dyes in the classes of oxazine derivatives (e.g.","MR 121, JA 242, JA 243) and rhodamines (e.g.","JA 165, JA 167, JA 169), which absorb and emit in the red region of the spectrum of visible light [see, for example, Müller R., Zander C., Sauer M., Deimel M., Ko D. S., Siebert S., Arden-Jacob J., Deltau G., Marx N. J., Drexhage K. H., Wolfrum J. Chem.","Phys.","Let.","1996, 262, 716-722] are particularly suitable.","These red dyes are extinguished by tryptophan by one to two orders of magnitude better than other dyes.","When red dyes are used there is a marked increase in sensitivity, because otherwise the autofluorescence of biologically relevant proteins, e.g.","blood sera and ascites, which mostly occurs in the green region of the visible spectrum, makes sensitive detection more difficult.","With the method as claimed in the invention, sensitive tests can also be carried out on these biologically relevant samples, even on undiluted blood sera.","The aim is also achieved by a second test for the detection of exopeptidases (enzyme).","Once again a peptide is prepared.","This time the peptide contains at least one group that contains a quencher and a fluorophore on different amino acids.","Once again the amino acid tryptophan is used as the quencher.","A fluorescent dye is again used as the fluorophore.","A fluorescent dye is chosen that can be coupled covalently to peptides or proteins.","Furthermore, the fluorescence of the dye can be extinguished by tryptophan.","The sequence of the peptide must be selected in such a way that the quencher and the fluorophore are spatially sufficiently close together, so long as the amino acids carrying the quencher or the fluorescent dye have not yet been separated by the enzyme, to ensure the possibility of an at least partial extinction of the emission of the fluorophore.","Moreover, after separation of the amino acids carrying the quencher or the fluorescent dye by the enzyme, a sufficient spatial distance must be provided between any other quencher and the fluorophore, to achieve an at least partial increase in emission of the fluorophore.","The peptide prepared in this way is mixed in a solution with the enzyme to be detected.","The intensity of the emission of the fluorophore in the solution is determined.","In many aspects this test corresponds to that already described for detecting an endopeptidase.","It is, however, easier to detect an exopeptidase.","The exopeptidase digests the protein or peptide from its end.","It detaches one amino acid after another.","A tryptophan is also detached, among others.","The dye coupled to an amino acid in the vicinity is therefore no longer extinguished, and there is an increase in fluorescence.","The same applies when the amino acid is detached first with the dye.","If the substrate contains a multiplicity of tryptophan-dye groups, an increase in fluorescence can be observed repeatedly.","The effect is then a multiple increase in fluorescence.","Once again, the aforementioned red dyes can be used advantageously.","The aim is finally achieved by a third test for the detection of endopeptidases (enzyme).","This test again has many similarities and some decisive differences.","First a peptide is prepared.","A group that prevents an exopeptidase from digesting the peptide is arranged at one end of the peptide.","The group at the end of the peptidase that prevents the digestion of the peptide by an exopeptidase is also called a stop group or stop molecule.","This can be, for example, D-amino acids, DNA or PNA, which are not cleaved by the exopeptidase.","Another possibility for providing a stop group is for an amino acid or amino acid sequence that is not cleaved by the exopeptidase to be arranged at the end of the peptide, by an appropriate choice of peptide sequence.","The carboxypeptidase A as exopeptidase, for example, does not cleave beyond the amino acid proline at the end of a peptide.","The arrangement of an ordinary amino acid as stop group facilitates synthesis of the peptide.","The exopeptidase can thus be present in solution with the peptide or protein, without the latter being attacked.","In addition the peptide contains an identification sequence of the endopeptidase that is to be detected and can therefore serve as substrate for the enzyme, yielding a cleavage site between two amino acids in the identification sequence in the peptide.","Viewed from the stop group in the sequence of the peptide on the other side of the cleavage site, the peptide contains at least one group that contains a quencher and a fluorophore on different amino acids.","The peptide sequence is chosen such that spatially the quencher and fluorophore are sufficiently close together, so long as the amino acids carrying the quencher and the fluorescent dye have not yet been separated by an exopeptidase, to ensure the possibility of an at least partial extinction of the emission of the fluorophore.","Moreover, it is necessary to ensure that after separation of the amino acids carrying the quencher and the fluorescent dye by an exopeptidase, a sufficient spatial distance is provided between any other quencher and the fluorophore, to achieve an at least partial increase in emission of the fluorophore.","Finally, the peptide thus prepared, the at least one exopeptidase and the endopeptidase that is to be detected are mixed in a solution.","This results in binding between peptide as substrate and endopeptidase as enzyme.","The peptide is cleaved at the cleavage site within the molecule.","As a result the protective stop group at the end of the substrate is separated from the rest of the substrate.","The residue that remains then no longer has a protective stop group, and an exopeptidase can digest the rest of the substrate.","Such an exopeptidase can be added to the solution in sufficient quantity, together with the substrate.","It can thus be used as a kit together with the substrate.","By digestion by means of the exopeptidase, the quencher and fluorophore are separated.","There is again an increase in fluorescence.","Amino acid sequences of any length can be present between the stop group and the region of the peptide or protein carrying the quencher-and-fluorophore groups, without impairing the sensitivity of detection.","It is thus also possible to detect enzymes that possess a long identification sequence, such as HIV protease.","If a carboxypeptidase is used as exopeptidase, the stop group is placed at the C-terminal end of the peptide, so that subsequent digestion by the carboxypeptidase can take place.","If, however, an aminopeptidase is used as exopeptidase, the stop group is placed at the N-terminal end of the peptide.","If the substrate contains a multiplicity of quencher-and-fluorophore groups, a multiple increase in fluorescence can again be observed.","The measurement time can be shortened considerably as a result.","There is at the same time a marked increase in the signal.","The great advantage of multiple marking is that with a single cleavage and thus with a single endopeptidase to be detected, not only one, but many fluorophores are activated.","As a result we obtain a more definite increase in signal in a much shorter time.","This means that the increase in signal can even be measured with less sensitive instruments.","There is also a shift of the limit of detection to lower values.","This test also detects endopeptidases specifically, with the other advantages that have already been mentioned.","Various dyes that are bound covalently to an amino acid can be selected as the fluorophore.","A quencher can also be bound covalently to an amino acid.","A suitable combination is for example the quencher DABCYL or guanosine and as fluorophore, for example, the dyes fluorescein, coumarin or eosins.","For this third test, once again the amino acid tryptophan can be used as quencher, together with a fluorescent dye as fluorophore.","It must be possible for the fluorescent dye to couple covalently to peptides or proteins, and for its fluorescence to be extinguished by tryptophan.","The advantages of this choice have already been explained.","Once again it is possible to use red dyes.","Among other things, for this third test there are yet other possibilities for producing the quencher/fluorophore pair.","The quencher can be at least one first fluorescent dye and the fluorophore at least one second fluorescent dye, provided they can both be coupled covalently to peptides or proteins.","What is important is that the two dyes can extinguish one another when they are spatially close together.","This can result from dimer formation or other mechanisms, such as energy transfer.","After removal of the stop group by the endopeptidase that is to be detected, the exopeptidase digests the rest of the peptide chain, so that the dyes become farther apart and quenching is thus no longer possible.","The resulting increase in intensity of fluorescence is measured.","Preferably all the aforementioned rhodamines and oxazines are suitable as dyes for this, and in addition cyanine derivatives (e.g.","Cy 3, Cy 5, Cy 7, obtainable from Amersham Biosciences), all BODIPY dyes, all coumarins, fluorescein and Texas Red.","The invention is explained in more detail below on the basis of examples of application, shown schematically in the diagrams.","Reference numbers in the individual diagrams denote the same items.","FIG.","1 is a schematic representation of the basic principle of the detection reaction; FIG.","2 is a schematic representation of the processes in photo-induced electron transfer; FIG.","3 is a schematic representation of the basic principle of a variant of the detection reaction; FIG.","4 is a schematic representation of the detection reaction as claimed in FIG.","4 with multiple marking; and FIG.","5 shows the structural formulae of the dyes mentioned.","First Test Detection of an Endopeptidase FIG.","1 is a schematic representation of the principle of detection of an endopeptidase, which is represented in FIG.","1 by the scissors symbol.","The peptide, consisting of four amino acids (AS) and tryptophan (Trp), serves as substrate.","A dye molecule (Dye), preferably MR 121, is coupled to the second amino acid from the left.","Possibilities for coupling are described in [Bodanszky M.: “Peptide Chemistry: A Practical Textbook”, Springer Verlag 1986] [Anderson G. W., Zimmerman J. E., Callahlan F. M.: “The use of esters of N-Hydroxysuccinimide in Peptide Synthesis”, J.","Am.","Chem.","Soc.","1964, 86, 1839-1842].","If, for example, we would like to detect the endopeptidase trypsin, which hydrolyzes the peptide bond between two arginines, the following peptide can be synthesized as substrate: Gln-Lys(MR 121)-Arg-Arg-Trp, as shown schematically in FIG.","1.Tryptophan acts as quencher on the dye.","The mechanism of quenching is static quenching, in contrast to a dynamic impact quenching.","The dye forms a complex with tryptophan, called a ground-state complex, which hardly fluoresces.","In this ground-state complex there is electron transfer from tryptophan to the excited dye molecule.","Electron transfer will be explained briefly in the following, referring to FIG.","2.This depicts fluorescence quenching of the excited dye molecule F* by a tryptophan residue W. The black circles represent electrons.","The HOMO (highest occupied molecular orbital) and the LUMO (lowest unoccupied molecular orbital) are shown in each case.","The HOMO is the energetically highest molecular orbital occupied in the electronic ground state.","The LUMO is the energetically lowest molecular orbital, unoccupied in the electronic ground state; it is as a rule the molecular orbital that is occupied in the first excited state.","Basically there are two possibilities for fluorescence quenching by photo-induced electron transfer.","In the case shown on the left in FIG.","2, the tryptophan residue W acts as an electron donor (Donor).","Following excitation of the fluorophore F*, an electron is transferred from the doubly occupied HOMO of the tryptophan residue W to the now singly occupied HOMO of the fluorophore F* (path 1).","There is reduction of the excited fluorophore F* by the tryptophan residue W. The electron in the LUMO of fluorophore F* can then be transferred to the now singly occupied HOMO of tryptophan residue W (path 2).","This case occurs between red dyes like MR 121 and tryptophan.","In the case shown on the right in FIG.","2, the tryptophan residue W acts as an electron acceptor (Acceptor).","The electron migrates from the singly occupied LUMO of the excited fluorophore F* to the unoccupied LUMO of tryptophan residue W (path 3).","There is oxidation of the excited fluorophore F* by the tryptophan residue W. The electron in the LUMO of the tryptophan residue W can then return to the HOMO of the fluorophore (path 4).","In both cases, after electron transfer the electron can no longer return to the HOMO from the LUMO of the excited fluorophore F* by emitting a photon.","The first excited state has been deactivated without radiation.","The fluorescence is quenched.","With the red dyes under examination, tryptophan always acts as an electron donor.","This knowledge makes it possible, for example, to choose suitable dyes on the basis of their electrochemically determined potentials.","The fluorescence signals are preferably detected in solution in a cuvette in conventional fluorescence spectrometers.","This is generally called “homogeneous assay”.","More sensitive measurements can be carried out using confocal spectroscopy.","Instead of the fluorescence intensity, it is also possible to use the fluorescence lifetime for detection.","The fluorescence lifetime is shorter in the quenched state than in the free, unquenched state.","Another possibility for detecting a quenched or unquenched state is measurement of changes in polarization.","The polarization properties of emissions, e.g.","fluorescence signals, can vary between a quenched and an unquenched state.","In the second and third test it is also possible to use FRET systems, i.e.","pairs of dyes that extinguish each other by energy transfer.","These often show different wavelengths in the quenched and unquenched state, which can be used for detection.","As excitation light sources it is preferable to use diode lasers, otherwise any other lasers or lamps with suitable wavelength.","It is preferable to use pulsed diode lasers for determining the extinction time of the fluorophores.","As well as the so-called homogeneous assay, there is also a so-called heterogeneous assay.","This generally denotes detection on a surface.","The dye-marked peptides or proteins can be bound covalently to a surface at a C-end, an N-end or via amino acid residues (e.g.","cysteine or lysine).","Possible surfaces are for example modified glass surfaces (accordingly chips or biochips) or (possibly magnetic) beats.","For this purpose the surfaces can be coated e.g.","with linear and/or crosslinked polyethyleneglycols or hyaluronic acid, to prevent adsorption of the molecules.","If an amino acid, by which the peptide is bound to the surface, is located on the same side of the cleavage site as the dye, cleavage through the enzyme that is to be detected has the effect that the dye bound covalently to the surface stays back and its fluorescence can no longer be extinguished there.","The fluorescence can then be detected evanescently or by means of surface screenings.","It is also possible to immobilize the peptide on the surface in such a way that the enzyme actually disrupts the bond between dye and surface as a result of the cleavage.","The fluorescence of the unquenched dye can then be detected in the solution.","Second Test Detection of an Exopeptidase The second test described at the beginning for detecting an exopeptidase is carried out entirely similarly to the first test.","The differences were described at the beginning.","Third Test Detection of an Endopeptidase FIG.","3 is a schematic representation of the principle of detection of an endopeptidase as claimed in the third test.","The endopeptidase to be detected is represented in FIG.","3A by the scissors symbol 10.The peptide, consisting of a stop group (Stop), two amino acids (AS), tryptophan (Trp) and another amino acid (AS), to which a dye molecule (Dye) is coupled, preferably MR 121, serves as substrate.","The endopeptidase to be detected 10 cleaves the peptide between the two adjacent amino acids AS.","This splits off the stop group (Stop) together with an amino acid AS (see FIG.","3B).","This makes it possible for the exopeptidase 12, which is also present in the solution and has up till now been prevented from digestion by the stop group (Stop), to digest the residual peptide.","The exopeptidase 12 successively splits off first an amino acid AS and then tryptophan (Trp).","As a result of this, the dye (Dye) on the amino acid that remains (AS) is no longer located spatially close to the tryptophan, because the tryptophan diffuses away in the solution.","Therefore the dye is no longer extinguished by tryptophan.","Its fluorescence can be detected as a signal increase in the spectrometer.","For detecting HIV protease, the following peptide can be used: (N terminus) Lys(MR 121)-Trp-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Pro-Pro (C terminus).","This peptide contains the identification sequence for cleavage by HIV protease.","This peptide can be used in combination with carboxypeptidase A, which does not cleave beyond proline.","Therefore the two proline residues at the C terminus of the above peptide serve as a stop group.","HIV protease cleaves between tyrosine and proline.","Two fragments are produced: Lys(MR 121)-Trp-Ser-Gln-Asn-Tyr-OH and Pro-Ile-Val-Gln-Pro-Pro.","The stop group is removed thereby and carboxypeptidase A can digest the first fragment counting from the C terminus.","This also separates the dye-marked lysine and tryptophan.","An increase in fluorescence can be detected.","FIG.","4 shows the third test with signal intensification by multiple marking.","FIG.","4A shows the peptide, as in FIG.","3A, but with an alternating succession of tryptophan (Trp) and a dye-marked amino acid (AS+Dye).","If the endopeptidase to be detected 10 splits off the stop group (Stop) (see FIG.","4B), the exopeptidase 12 can digest the peptide successively.","In the process, tryptophan and free amino acid with the bound dye (AS+Dye) are released successively (see FIG.","4C).","Thus, with one cleavage of the endopeptidase, a large number of dyes is released, whose emission is no longer extinguished.","There is accordingly a rapid and strong signal increase.","This shortens the measurement time, even for highly sensitive measurements, to minutes or seconds.","Finally, FIG.","5 shows the structure of the preferred dyes used, as mentioned at the beginning."]],"string":"[\n [\n \"Specific detection of proteolytic enzymes\",\n \"Specific detection of proteolytic enzymes is achieved by extinguishing dye fluorescence by the amino acid tryptophan.\",\n \"Tryptophan is disposed on one side of the cutting site of a proteolytic enzyme while an amino acid marked with a dye is arranged on the other side.\",\n \"Extinction of fluorescence occurs prior to enzyme cutting.\",\n \"Spatial separation of the tryptophan and the dye takes place after cutting, whereby fluorescence extinction does not occur.\",\n \"The dye can then fluoresce and a signal increase occurs thereby indicating that cutting has been carried out and the presence of the enzyme.\"\n ],\n [\n \"1.\",\n \"(canceled) 2.The peptide as claimed in claim 21, wherein the fluorescent dye is one of an oxazine derivative or a rhodamine derivative.\",\n \"3.A method for the detection of an endopeptidase comprising the following steps: mixing the peptide as claimed in claim 21 and an enzyme to be detected in a solution; and determining an intensity of emission of the fluorophore in the solution.\",\n \"4.A device on which a peptide as claimed in claim 21 is immobilized.\",\n \"5.A device for immobilizing the peptide as claimed in claim 21, wherein the peptide is immobilized on the device in such a way that after cleavage by the enzyme to be detected, the fluorophore remains immobilized on a surface, whereas the quencher is no longer immobilized on the surface.\",\n \"6.\",\n \"(canceled) 7.The peptide as claimed in claim 22, wherein the fluorescent dye is an oxazine derivative or a rhodamine derivative.\",\n \"8.The peptide as claimed in claim 22, wherein: the quencher is at least one first fluorescent dye; the fluorophore is at least one second fluorescent dye; the at least one first and one second fluorescent dye can couple covalently to peptides or proteins; and the at least one first and second fluorescent dye can quench each other.\",\n \"9.A method for the detection of an exopeptidase comprising the following steps: mixing a peptide as claimed in claim 22 and the enzyme to be detected in a solution; and determining the intensity of emission of the fluorophore in the solution.\",\n \"10.A device on which a peptide as claimed in claim 22 s immobilized.\",\n \"11.A device for immobilizing the peptide as claimed in claim 22, wherein the peptide is immobilized on the device in such a way that after cleavage by the enzyme to be detected, the fluorophore remains immobilized on the surface, whereas the quencher is no longer immobilized on the surface.\",\n \"12.\",\n \"(canceled) 13.\",\n \"(canceled) 14.The peptide as claimed in claim 24, wherein the fluorescent dye is an oxazine derivative or a rhodamine derivative.\",\n \"15.The peptide as claimed in claim 23, wherein the quencher is at least one first fluorescent dye; the fluorophore is at last one second fluorescent dye; the at least one first and second fluorescent dye can couple covalently to peptides or proteins; and the at least one first and second fluorescent dye can quench each other, 16.\",\n \"(canceled) 17.A method for the detection of an endopeptidase comprising the following steps; mixing a peptide as claimed in claim 23 at least one exopeptidase and the endopeptidase to be detected, in a solution; and determining the intensity of emission of the fluorophore in the solution.\",\n \"18.\",\n \"(canceled) 19.\",\n \"(canceled) 20.Kit comprising the peptide as claimed in claim 23 and including at least one exopeptidase.\",\n \"21.A peptide with the following properties, the peptide comprising an identification sequence of an endopeptidase and functioning as substrate for the endopeptidase and providing a cleavage site between two amino acids at the identification sequence in the peptide; and at least one fluorophore in the form of a fluorescent dye for covalently coupling to peptides or proteins and located at one side of the cleavage site; and at least one quencher in the form of tryptophan arranged on the other side of the cleavage site; and wherein the quencher is sufficiently proximate to the fluorophore so that before the substrate at the cleavage site between quencher and fluorophore is cleaved, at least a partial quenching of a fluorescent emission from the fluorophore is realized; and wherein after the substrate is cleaved by the endopeptidase at the cleavage site, emission by the fluorophore is increased.\",\n \"22.A peptide with the following properties, comprising: an amino acid sequence, and at least one group containing tryptophan as a quencher and a fluorescent dye as a fluorophore on different amino acids; wherein the fluorescent dye is able to couple covalently to peptides or proteins and to emit fluorescence that is quenched by tryptophan; wherein the sequence contains sufficient spatial proximity between the quencher and the fluorophore so that as long as the amino acids carrying the quencher and the fluorescent dye have not been separated by an exopeptidase, at least a partial quenching of the emitted fluorescence of the fluorophore is realized; and wherein after the amino acids carrying the quencher and the fluorescent dye are separated by the exopeptidase, at least a partial increase in fluorescent emission of the fluorophore is realized.\",\n \"23.A peptide with the following properties, the peptide comprising: an identification sequence of an endopeptidase and functioning as substrate for the endopeptidase and providing a cleavage site between two amino acids at the identification sequence in the peptide; and a stop group that prevents an exopeptidase from digesting the peptide and arranged at one end of the peptide; and wherein the identification sequence includes at least one group containing a quencher and a fluorophore on different amino acids and arranged downstream from the stop group; wherein the quencher and the fluorophore are sufficiently proximate, so that as long as the amino acids carrying the quencher and the fluorescent dye have not yet been separated by an exopeptidase, at least a partial quenching of the emission of the fluorophore is realized; and wherein after the amino acids carrying the quencher and the fluorescent dye are separated by an exopeptidase, a spatial distance is realized between the quencher or any other quencher and the fluorophore sufficient to realize an at least partial increase in emission the fluorophore.\",\n \"24.The peptide as claimed in claim 23, wherein the fluorophore is a fluorescent dye for coupling covalently to peptides or proteins and the quencher is tryptophan for quenching the fluorescence emitted from the fluorescent dye.\"\n ],\n [\n \"Proteolytic enzymes are proteases or peptidases, i.e.\",\n \"enzymes that are able to split or lyse peptides or proteins.\",\n \"The word “peptide” generally denotes a shorter amino acid sequence, whereas the word “protein” generally denotes larger molecules consisting of an amino acid sequence.\",\n \"Both terms are used synonymously in the following.\",\n \"Peptidases and proteases play a decisive role in protein activation, cell regulation and signal transmission.\",\n \"Their detection at maximum possible sensitivity is therefore of enormous importance for understanding the functioning of a cell and its communication, and for monitoring the course of diseases.\",\n \"Furthermore, precise knowledge of peptidase or protease concentration permits the targeted development of new therapeutic agents for inhibition, e.g.\",\n \"of HIV protease.\",\n \"HIV protease breaks down the HIV protein, which is at first very long after multiplication in a cell, into functioning individual proteins, which only then permit the further activity and growth of the HIV virus.\",\n \"There are two possible approaches.\",\n \"Firstly, if the HIV protease is blocked, the HIV virus cannot replicate and its propagation is halted.\",\n \"Secondly, infection with the HIV virus can take place via detection of the HIV protease.\",\n \"Specific detection of HIV protease can therefore serve as an AIDS test.\",\n \"Moreover, protease tests are attracting increasing attention in medical research, because more and more diseases, including cancers, are being connected with enzymes.\",\n \"Tumor-associated proteases are increasingly the object of oncological-biochemical research.\",\n \"These proteases bring about the breakdown of proteins of the tumor stroma and of the basal membrane and thus permit tumor cell invasion.\",\n \"Therefore investigations of protease systems that are overexpressed in tumor cells are among the new prognosis factors in tumor diagnostics.\",\n \"Another important aspect is the use of protease tests in quality control of biochemical products.\",\n \"If, for example proteins, enzymes, peptides, etc.\",\n \"are being sold, it is necessary to ensure that the products sold are free from proteases, so that the product sold does not break down on its own.\",\n \"The same can be used for testing the specific activity of proteases, e.g.\",\n \"after prolonged storage, regarding the question as to whether the enzymes are still active.\",\n \"The proteolytic enzymes are divided into endopeptidases (proteases) and exopeptidases.\",\n \"Endopeptidases cleave amino linkages inside peptides.\",\n \"Exopeptidases digest peptides amino acid by amino acid starting from their ends, i.e.\",\n \"they cleave terminal amino acids.\",\n \"The exopeptidases can be further subdivided into aminopeptidases and carboxypeptidases, as claimed in their activity at the amino or N end or at the carboxy or C end of the peptide.\",\n \"For determination of the concentration or activity of various peptidases and proteases, to date various fluorescence-based tests have been developed, and their operating principles and sensitivities are briefly described below.\",\n \"One possibility for detecting proteolytic enzymes is offered by the “EnzChek Protease Assay Kit” from the company Molecular Probes (Eugene, USA).\",\n \"For detecting proteases, this kit employs casein derivatives, to which very many pH-insensitive green or red fluorescing BODIPY dyes are coupled (Karolin J., Johansson B.\",\n \"A., Strandberg L., Ny T.; J.\",\n \"Am.\",\n \"Chem.\",\n \"Soc., 1994, 116, 7801-7806).\",\n \"On account of the very high degree of marking of the protein, the intermolecular distances between the fluorophores are very small (typically of the order of a few nanometers).\",\n \"The dyes therefore extinguish each other, inter alia by dimer formation.\",\n \"Casein is a large enzyme, which contains many binding sites for proteases, because it has many different sequence segments.\",\n \"If the dye-marked casein derivative comes into contact with proteases, e.g.\",\n \"with trypsin, which hydrolyzes peptide bonds of the basic amino acids arginine and lysine on the carboxy side, these split the casein derivative into smaller peptides, so that as a rule the distance between the dyes increases.\",\n \"Extinction does not occur, and an increase in fluorescence is observed, indicating that proteolytic enzymes are present.\",\n \"For trypsin, for example, the detection sensitivity of this kit is a few ng/ml.\",\n \"For these small quantities of protease the detection time or measurement time is more than 10 hours.\",\n \"This test detects many proteolytic enzymes nonspecifically, since casein is digested by elastase, pepsin, thermolysin, papain, trypsin, etc.\",\n \"An enzyme test based on Rhodamine 110 for various proteases and peptidases (Leytus S. P., Melhado L. L., Mangel W. F.: Rhodamine-based compounds as fluorogenic substrates for serine protease, (1983) Biochem.\",\n \"J., 209 (2): 299-307; Leytus S. P., Patterson W. L., Mangel W. F.: New class of sensitive and selective fluorogenic substrates for serine proteinases.\",\n \"Amino acid and dipeptide derivatives of rhodamine, (1983) Biochem.\",\n \"J., 215 (2): 253-260) is also being marketed for the detection of serine proteases.\",\n \"In this test, at least one of the amino groups of Rhodamine 110 is coupled covalently to an amino acid (usually arginine), causing strong extinction of the fluorescence of the fluorophore.\",\n \"After digestion of the peptide bonds there is a dramatic increase in fluorescence of Rhodamine 110.For caspase-3, the sensitivity of the test is a few ng absolute quantity of enzyme.\",\n \"Disadvantages are that the principle is not of general applicability, sometimes its action is nonspecific, and it is relatively insensitive.\",\n \"A protease or peptidase test can also be accomplished by means of Forster fluorescence resonance energy transfer (FRET) (Forster Th.\",\n \": Zwischenmolekulare Energiewanderung und Fluoreszenz [Intermolecular Energy Transfer and Fluorescence], (1948) Annalen der Physik, 2: 55-75).\",\n \"In this, the specific identification sequence of an endopeptidase is marked covalently at both ends with a donor dye and an acceptor dye.\",\n \"Owing to spectral overlap of donor emission with acceptor absorption, the fluorescence of the donor is extinguished at small distances.\",\n \"If the peptide is cleaved by the endopeptidase, the spatially close contact between donor and acceptor is lost, leading to an increase in fluorescence.\",\n \"Detection of HIV protease will be briefly explained here, as an example.\",\n \"The specific cleaving sequence of HIV protease is coupled, near the cleavage site, with an EDANS dye (donor) and a DABCYL dye (acceptor).\",\n \"The peptide sequence is then: Arg-Glu(EDANS)-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys-(DABCYL)-Arg, using the 3-letter abbreviations for the amino acids.\",\n \"After cleavage by the HIV protease we have two parts: Arg-Glu(EDANS)Ser-Gln-Asn-Tyr-OH and Pro-Ile-Val-Gln-Lys(DABCYL)-Arg.\",\n \"The principle is of general applicability, but there must be a suitable small distance between the two dyes.\",\n \"Sensitivity is of the order of nanomolar solutions of HIV protease.\",\n \"A disadvantage is the expensive synthesis of the substrate with specific coupling of a donor and an acceptor.\",\n \"If the substrate is to some extent marked incompletely, e.g.\",\n \"with just one donor, this leads to poor sensitivity of the test.\",\n \"The aim of the invention is to improve the possibilities for a specific detection of proteolytic enzymes.\",\n \"This aim is achieved by the inventions as claimed in the independent claims.\",\n \"Advantageous developments are described in the subclaims.\",\n \"A first method of detection (“test” for short) is used for the detection of endopeptidases (also briefly referred to as enzymes hereinafter) by means of special peptides.\",\n \"For this purpose, a peptide is prepared with the properties enumerated below.\",\n \"The peptide contains an identification sequence of the endopeptidase that is to be detected, and can therefore serve as substrate for the enzyme, with a cleavage site between two amino acids in the identification sequence in the peptide.\",\n \"The peptide contains at least one quencher and at least one fluorophore.\",\n \"“Quenching” means reduction of the emission of the fluorophore due to interaction with the aforesaid quencher.\",\n \"A fluorescent dye is used as the fluorophore.\",\n \"Quenching causes a decrease in fluorescence quantum yield.\",\n \"The words “extinction” and “extinguishing molecule” or “extinguisher” are also used as synonyms for “quenching” and “quencher”.\",\n \"The amino acid tryptophan is used as quencher.\",\n \"Other amino acids, such as tyrosine or histidine, also exhibit slight fluorescence extinction, but do not have the strong quenching action of tryptophan.\",\n \"The fluorescent dye is selected on the basis that its fluorescence can be extinguished by tryptophan.\",\n \"Furthermore, it should be possible for the fluorescent dye to be coupled covalently to peptides or proteins.\",\n \"As a rule the fluorophore is coupled to an amino acid.\",\n \"Attainment of high sensitivity of the test requires targeted selection of the peptide sequence: the quencher must be arranged on one side of the cleavage site, with the fluorophore on the other side of the cleavage site.\",\n \"The sequence must be selected so that the quencher and the fluorophore are sufficiently spatially close to one another, so long as the substrate has still not been cleaved by the enzyme at the cleavage site between quencher and fluorophore, to provide the possibility of at least partial extinction of the emission of the fluorophore.\",\n \"After cleavage of the substrate by the enzyme at the cleavage site between quencher and fluorophore, a definite increase in emission of the fluorophore must be possible.\",\n \"Thus, the peptide or protein is not marked with the fluorescent dye on the tryptophan itself, but on another amino acid, which is separated from the tryptophan by the cleavage site.\",\n \"Furthermore, an amino acid other than tryptophan is chosen for coupling the fluorescent dye, e.g.\",\n \"lysine, arginine or cysteine.\",\n \"The fluorescent dye can also be coupled to one end of the peptide or protein.\",\n \"The largest increase in fluorescence is obtained if the tryptophan and the dye-marked amino acid are located directly on either side of the cleavage site.\",\n \"For dye MR 121, for example, an increase in fluorescence by a factor of 10 can be observed after cleavage of the peptide by the enzyme.\",\n \"Moreover, no tryptophan is to be arranged in adjacent positions on the same side of the cleavage site of the peptide sequence as the dye, and especially not in immediately adjacent positions, so that the dye is not quenched by this additional tryptophan.\",\n \"Appropriate choice of substrate sequence makes it possible to mark the substrate with just one fluorescent dye, and not with at least two, as was necessary until now.\",\n \"This simplifies the synthesis of the substrate.\",\n \"In addition there is less disturbance of binding of the enzyme to the substrate by coupled dyes, resulting in greater specificity of binding between substrate and enzyme.\",\n \"Finally, the peptide thus prepared and the enzyme to be detected are mixed in a solution.\",\n \"If binding occurs between peptide and peptidase, the peptide is cleaved at the cleavage site.\",\n \"As a result, the spatial distance between the fluorophore and the quencher becomes greater, and the emission of the fluorophore increases.\",\n \"This increase in emission is determined and serves for detecting those peptidases or proteases that bind to the identification sequence and can cleave the peptide there.\",\n \"Accordingly, the detection reaction is specific.\",\n \"It can be directed towards the detection of specific enzymes by varying the peptide sequence.\",\n \"In particular, the test can detect enzymes specifically, even when other types of enzymes are also present in the solution.\",\n \"For example, the enzyme trypsin can be detected in a solution that also contains elastase and chymotrypsin.\",\n \"If the temporal course of the increase in emission is traced, the concentration of the enzyme can be determined quantitatively.\",\n \"Using the test as claimed in the invention, it is even possible to detect proteases or peptidases at a concentration of 10{circumflex over ( )}−15 mol/l.\",\n \"As dyes, consideration may be given to all fluorescent dyes that are measurably extinguished in their fluorescence by the amino acid tryptophan.\",\n \"Dyes in the classes of oxazine derivatives (e.g.\",\n \"MR 121, JA 242, JA 243) and rhodamines (e.g.\",\n \"JA 165, JA 167, JA 169), which absorb and emit in the red region of the spectrum of visible light [see, for example, Müller R., Zander C., Sauer M., Deimel M., Ko D. S., Siebert S., Arden-Jacob J., Deltau G., Marx N. J., Drexhage K. H., Wolfrum J. Chem.\",\n \"Phys.\",\n \"Let.\",\n \"1996, 262, 716-722] are particularly suitable.\",\n \"These red dyes are extinguished by tryptophan by one to two orders of magnitude better than other dyes.\",\n \"When red dyes are used there is a marked increase in sensitivity, because otherwise the autofluorescence of biologically relevant proteins, e.g.\",\n \"blood sera and ascites, which mostly occurs in the green region of the visible spectrum, makes sensitive detection more difficult.\",\n \"With the method as claimed in the invention, sensitive tests can also be carried out on these biologically relevant samples, even on undiluted blood sera.\",\n \"The aim is also achieved by a second test for the detection of exopeptidases (enzyme).\",\n \"Once again a peptide is prepared.\",\n \"This time the peptide contains at least one group that contains a quencher and a fluorophore on different amino acids.\",\n \"Once again the amino acid tryptophan is used as the quencher.\",\n \"A fluorescent dye is again used as the fluorophore.\",\n \"A fluorescent dye is chosen that can be coupled covalently to peptides or proteins.\",\n \"Furthermore, the fluorescence of the dye can be extinguished by tryptophan.\",\n \"The sequence of the peptide must be selected in such a way that the quencher and the fluorophore are spatially sufficiently close together, so long as the amino acids carrying the quencher or the fluorescent dye have not yet been separated by the enzyme, to ensure the possibility of an at least partial extinction of the emission of the fluorophore.\",\n \"Moreover, after separation of the amino acids carrying the quencher or the fluorescent dye by the enzyme, a sufficient spatial distance must be provided between any other quencher and the fluorophore, to achieve an at least partial increase in emission of the fluorophore.\",\n \"The peptide prepared in this way is mixed in a solution with the enzyme to be detected.\",\n \"The intensity of the emission of the fluorophore in the solution is determined.\",\n \"In many aspects this test corresponds to that already described for detecting an endopeptidase.\",\n \"It is, however, easier to detect an exopeptidase.\",\n \"The exopeptidase digests the protein or peptide from its end.\",\n \"It detaches one amino acid after another.\",\n \"A tryptophan is also detached, among others.\",\n \"The dye coupled to an amino acid in the vicinity is therefore no longer extinguished, and there is an increase in fluorescence.\",\n \"The same applies when the amino acid is detached first with the dye.\",\n \"If the substrate contains a multiplicity of tryptophan-dye groups, an increase in fluorescence can be observed repeatedly.\",\n \"The effect is then a multiple increase in fluorescence.\",\n \"Once again, the aforementioned red dyes can be used advantageously.\",\n \"The aim is finally achieved by a third test for the detection of endopeptidases (enzyme).\",\n \"This test again has many similarities and some decisive differences.\",\n \"First a peptide is prepared.\",\n \"A group that prevents an exopeptidase from digesting the peptide is arranged at one end of the peptide.\",\n \"The group at the end of the peptidase that prevents the digestion of the peptide by an exopeptidase is also called a stop group or stop molecule.\",\n \"This can be, for example, D-amino acids, DNA or PNA, which are not cleaved by the exopeptidase.\",\n \"Another possibility for providing a stop group is for an amino acid or amino acid sequence that is not cleaved by the exopeptidase to be arranged at the end of the peptide, by an appropriate choice of peptide sequence.\",\n \"The carboxypeptidase A as exopeptidase, for example, does not cleave beyond the amino acid proline at the end of a peptide.\",\n \"The arrangement of an ordinary amino acid as stop group facilitates synthesis of the peptide.\",\n \"The exopeptidase can thus be present in solution with the peptide or protein, without the latter being attacked.\",\n \"In addition the peptide contains an identification sequence of the endopeptidase that is to be detected and can therefore serve as substrate for the enzyme, yielding a cleavage site between two amino acids in the identification sequence in the peptide.\",\n \"Viewed from the stop group in the sequence of the peptide on the other side of the cleavage site, the peptide contains at least one group that contains a quencher and a fluorophore on different amino acids.\",\n \"The peptide sequence is chosen such that spatially the quencher and fluorophore are sufficiently close together, so long as the amino acids carrying the quencher and the fluorescent dye have not yet been separated by an exopeptidase, to ensure the possibility of an at least partial extinction of the emission of the fluorophore.\",\n \"Moreover, it is necessary to ensure that after separation of the amino acids carrying the quencher and the fluorescent dye by an exopeptidase, a sufficient spatial distance is provided between any other quencher and the fluorophore, to achieve an at least partial increase in emission of the fluorophore.\",\n \"Finally, the peptide thus prepared, the at least one exopeptidase and the endopeptidase that is to be detected are mixed in a solution.\",\n \"This results in binding between peptide as substrate and endopeptidase as enzyme.\",\n \"The peptide is cleaved at the cleavage site within the molecule.\",\n \"As a result the protective stop group at the end of the substrate is separated from the rest of the substrate.\",\n \"The residue that remains then no longer has a protective stop group, and an exopeptidase can digest the rest of the substrate.\",\n \"Such an exopeptidase can be added to the solution in sufficient quantity, together with the substrate.\",\n \"It can thus be used as a kit together with the substrate.\",\n \"By digestion by means of the exopeptidase, the quencher and fluorophore are separated.\",\n \"There is again an increase in fluorescence.\",\n \"Amino acid sequences of any length can be present between the stop group and the region of the peptide or protein carrying the quencher-and-fluorophore groups, without impairing the sensitivity of detection.\",\n \"It is thus also possible to detect enzymes that possess a long identification sequence, such as HIV protease.\",\n \"If a carboxypeptidase is used as exopeptidase, the stop group is placed at the C-terminal end of the peptide, so that subsequent digestion by the carboxypeptidase can take place.\",\n \"If, however, an aminopeptidase is used as exopeptidase, the stop group is placed at the N-terminal end of the peptide.\",\n \"If the substrate contains a multiplicity of quencher-and-fluorophore groups, a multiple increase in fluorescence can again be observed.\",\n \"The measurement time can be shortened considerably as a result.\",\n \"There is at the same time a marked increase in the signal.\",\n \"The great advantage of multiple marking is that with a single cleavage and thus with a single endopeptidase to be detected, not only one, but many fluorophores are activated.\",\n \"As a result we obtain a more definite increase in signal in a much shorter time.\",\n \"This means that the increase in signal can even be measured with less sensitive instruments.\",\n \"There is also a shift of the limit of detection to lower values.\",\n \"This test also detects endopeptidases specifically, with the other advantages that have already been mentioned.\",\n \"Various dyes that are bound covalently to an amino acid can be selected as the fluorophore.\",\n \"A quencher can also be bound covalently to an amino acid.\",\n \"A suitable combination is for example the quencher DABCYL or guanosine and as fluorophore, for example, the dyes fluorescein, coumarin or eosins.\",\n \"For this third test, once again the amino acid tryptophan can be used as quencher, together with a fluorescent dye as fluorophore.\",\n \"It must be possible for the fluorescent dye to couple covalently to peptides or proteins, and for its fluorescence to be extinguished by tryptophan.\",\n \"The advantages of this choice have already been explained.\",\n \"Once again it is possible to use red dyes.\",\n \"Among other things, for this third test there are yet other possibilities for producing the quencher/fluorophore pair.\",\n \"The quencher can be at least one first fluorescent dye and the fluorophore at least one second fluorescent dye, provided they can both be coupled covalently to peptides or proteins.\",\n \"What is important is that the two dyes can extinguish one another when they are spatially close together.\",\n \"This can result from dimer formation or other mechanisms, such as energy transfer.\",\n \"After removal of the stop group by the endopeptidase that is to be detected, the exopeptidase digests the rest of the peptide chain, so that the dyes become farther apart and quenching is thus no longer possible.\",\n \"The resulting increase in intensity of fluorescence is measured.\",\n \"Preferably all the aforementioned rhodamines and oxazines are suitable as dyes for this, and in addition cyanine derivatives (e.g.\",\n \"Cy 3, Cy 5, Cy 7, obtainable from Amersham Biosciences), all BODIPY dyes, all coumarins, fluorescein and Texas Red.\",\n \"The invention is explained in more detail below on the basis of examples of application, shown schematically in the diagrams.\",\n \"Reference numbers in the individual diagrams denote the same items.\",\n \"FIG.\",\n \"1 is a schematic representation of the basic principle of the detection reaction; FIG.\",\n \"2 is a schematic representation of the processes in photo-induced electron transfer; FIG.\",\n \"3 is a schematic representation of the basic principle of a variant of the detection reaction; FIG.\",\n \"4 is a schematic representation of the detection reaction as claimed in FIG.\",\n \"4 with multiple marking; and FIG.\",\n \"5 shows the structural formulae of the dyes mentioned.\",\n \"First Test Detection of an Endopeptidase FIG.\",\n \"1 is a schematic representation of the principle of detection of an endopeptidase, which is represented in FIG.\",\n \"1 by the scissors symbol.\",\n \"The peptide, consisting of four amino acids (AS) and tryptophan (Trp), serves as substrate.\",\n \"A dye molecule (Dye), preferably MR 121, is coupled to the second amino acid from the left.\",\n \"Possibilities for coupling are described in [Bodanszky M.: “Peptide Chemistry: A Practical Textbook”, Springer Verlag 1986] [Anderson G. W., Zimmerman J. E., Callahlan F. M.: “The use of esters of N-Hydroxysuccinimide in Peptide Synthesis”, J.\",\n \"Am.\",\n \"Chem.\",\n \"Soc.\",\n \"1964, 86, 1839-1842].\",\n \"If, for example, we would like to detect the endopeptidase trypsin, which hydrolyzes the peptide bond between two arginines, the following peptide can be synthesized as substrate: Gln-Lys(MR 121)-Arg-Arg-Trp, as shown schematically in FIG.\",\n \"1.Tryptophan acts as quencher on the dye.\",\n \"The mechanism of quenching is static quenching, in contrast to a dynamic impact quenching.\",\n \"The dye forms a complex with tryptophan, called a ground-state complex, which hardly fluoresces.\",\n \"In this ground-state complex there is electron transfer from tryptophan to the excited dye molecule.\",\n \"Electron transfer will be explained briefly in the following, referring to FIG.\",\n \"2.This depicts fluorescence quenching of the excited dye molecule F* by a tryptophan residue W. The black circles represent electrons.\",\n \"The HOMO (highest occupied molecular orbital) and the LUMO (lowest unoccupied molecular orbital) are shown in each case.\",\n \"The HOMO is the energetically highest molecular orbital occupied in the electronic ground state.\",\n \"The LUMO is the energetically lowest molecular orbital, unoccupied in the electronic ground state; it is as a rule the molecular orbital that is occupied in the first excited state.\",\n \"Basically there are two possibilities for fluorescence quenching by photo-induced electron transfer.\",\n \"In the case shown on the left in FIG.\",\n \"2, the tryptophan residue W acts as an electron donor (Donor).\",\n \"Following excitation of the fluorophore F*, an electron is transferred from the doubly occupied HOMO of the tryptophan residue W to the now singly occupied HOMO of the fluorophore F* (path 1).\",\n \"There is reduction of the excited fluorophore F* by the tryptophan residue W. The electron in the LUMO of fluorophore F* can then be transferred to the now singly occupied HOMO of tryptophan residue W (path 2).\",\n \"This case occurs between red dyes like MR 121 and tryptophan.\",\n \"In the case shown on the right in FIG.\",\n \"2, the tryptophan residue W acts as an electron acceptor (Acceptor).\",\n \"The electron migrates from the singly occupied LUMO of the excited fluorophore F* to the unoccupied LUMO of tryptophan residue W (path 3).\",\n \"There is oxidation of the excited fluorophore F* by the tryptophan residue W. The electron in the LUMO of the tryptophan residue W can then return to the HOMO of the fluorophore (path 4).\",\n \"In both cases, after electron transfer the electron can no longer return to the HOMO from the LUMO of the excited fluorophore F* by emitting a photon.\",\n \"The first excited state has been deactivated without radiation.\",\n \"The fluorescence is quenched.\",\n \"With the red dyes under examination, tryptophan always acts as an electron donor.\",\n \"This knowledge makes it possible, for example, to choose suitable dyes on the basis of their electrochemically determined potentials.\",\n \"The fluorescence signals are preferably detected in solution in a cuvette in conventional fluorescence spectrometers.\",\n \"This is generally called “homogeneous assay”.\",\n \"More sensitive measurements can be carried out using confocal spectroscopy.\",\n \"Instead of the fluorescence intensity, it is also possible to use the fluorescence lifetime for detection.\",\n \"The fluorescence lifetime is shorter in the quenched state than in the free, unquenched state.\",\n \"Another possibility for detecting a quenched or unquenched state is measurement of changes in polarization.\",\n \"The polarization properties of emissions, e.g.\",\n \"fluorescence signals, can vary between a quenched and an unquenched state.\",\n \"In the second and third test it is also possible to use FRET systems, i.e.\",\n \"pairs of dyes that extinguish each other by energy transfer.\",\n \"These often show different wavelengths in the quenched and unquenched state, which can be used for detection.\",\n \"As excitation light sources it is preferable to use diode lasers, otherwise any other lasers or lamps with suitable wavelength.\",\n \"It is preferable to use pulsed diode lasers for determining the extinction time of the fluorophores.\",\n \"As well as the so-called homogeneous assay, there is also a so-called heterogeneous assay.\",\n \"This generally denotes detection on a surface.\",\n \"The dye-marked peptides or proteins can be bound covalently to a surface at a C-end, an N-end or via amino acid residues (e.g.\",\n \"cysteine or lysine).\",\n \"Possible surfaces are for example modified glass surfaces (accordingly chips or biochips) or (possibly magnetic) beats.\",\n \"For this purpose the surfaces can be coated e.g.\",\n \"with linear and/or crosslinked polyethyleneglycols or hyaluronic acid, to prevent adsorption of the molecules.\",\n \"If an amino acid, by which the peptide is bound to the surface, is located on the same side of the cleavage site as the dye, cleavage through the enzyme that is to be detected has the effect that the dye bound covalently to the surface stays back and its fluorescence can no longer be extinguished there.\",\n \"The fluorescence can then be detected evanescently or by means of surface screenings.\",\n \"It is also possible to immobilize the peptide on the surface in such a way that the enzyme actually disrupts the bond between dye and surface as a result of the cleavage.\",\n \"The fluorescence of the unquenched dye can then be detected in the solution.\",\n \"Second Test Detection of an Exopeptidase The second test described at the beginning for detecting an exopeptidase is carried out entirely similarly to the first test.\",\n \"The differences were described at the beginning.\",\n \"Third Test Detection of an Endopeptidase FIG.\",\n \"3 is a schematic representation of the principle of detection of an endopeptidase as claimed in the third test.\",\n \"The endopeptidase to be detected is represented in FIG.\",\n \"3A by the scissors symbol 10.The peptide, consisting of a stop group (Stop), two amino acids (AS), tryptophan (Trp) and another amino acid (AS), to which a dye molecule (Dye) is coupled, preferably MR 121, serves as substrate.\",\n \"The endopeptidase to be detected 10 cleaves the peptide between the two adjacent amino acids AS.\",\n \"This splits off the stop group (Stop) together with an amino acid AS (see FIG.\",\n \"3B).\",\n \"This makes it possible for the exopeptidase 12, which is also present in the solution and has up till now been prevented from digestion by the stop group (Stop), to digest the residual peptide.\",\n \"The exopeptidase 12 successively splits off first an amino acid AS and then tryptophan (Trp).\",\n \"As a result of this, the dye (Dye) on the amino acid that remains (AS) is no longer located spatially close to the tryptophan, because the tryptophan diffuses away in the solution.\",\n \"Therefore the dye is no longer extinguished by tryptophan.\",\n \"Its fluorescence can be detected as a signal increase in the spectrometer.\",\n \"For detecting HIV protease, the following peptide can be used: (N terminus) Lys(MR 121)-Trp-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Pro-Pro (C terminus).\",\n \"This peptide contains the identification sequence for cleavage by HIV protease.\",\n \"This peptide can be used in combination with carboxypeptidase A, which does not cleave beyond proline.\",\n \"Therefore the two proline residues at the C terminus of the above peptide serve as a stop group.\",\n \"HIV protease cleaves between tyrosine and proline.\",\n \"Two fragments are produced: Lys(MR 121)-Trp-Ser-Gln-Asn-Tyr-OH and Pro-Ile-Val-Gln-Pro-Pro.\",\n \"The stop group is removed thereby and carboxypeptidase A can digest the first fragment counting from the C terminus.\",\n \"This also separates the dye-marked lysine and tryptophan.\",\n \"An increase in fluorescence can be detected.\",\n \"FIG.\",\n \"4 shows the third test with signal intensification by multiple marking.\",\n \"FIG.\",\n \"4A shows the peptide, as in FIG.\",\n \"3A, but with an alternating succession of tryptophan (Trp) and a dye-marked amino acid (AS+Dye).\",\n \"If the endopeptidase to be detected 10 splits off the stop group (Stop) (see FIG.\",\n \"4B), the exopeptidase 12 can digest the peptide successively.\",\n \"In the process, tryptophan and free amino acid with the bound dye (AS+Dye) are released successively (see FIG.\",\n \"4C).\",\n \"Thus, with one cleavage of the endopeptidase, a large number of dyes is released, whose emission is no longer extinguished.\",\n \"There is accordingly a rapid and strong signal increase.\",\n \"This shortens the measurement time, even for highly sensitive measurements, to minutes or seconds.\",\n \"Finally, FIG.\",\n \"5 shows the structure of the preferred dyes used, as mentioned at the beginning.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10474263"}}},{"rowIdx":792,"cells":{"text":{"kind":"list like","value":[["Modifiable assembly of microscopic apertures","The invention concerns a modifiable assembly of microscopic apertures comprising several plates (100, 110) coated with an opaque deposition except on transparent parts, for example (101, 114, 115), capable of sliding relative to one another, separated by an oil layer (117).","The invention is applicable to microscopic apertures for single-point or multiple-point confocal microscopy."],["1.A modifiable set of at least one pinhole intended for filtering a light beam, comprising several plates (100, 110) each carrying at least one pinhole, one of these plates (110) carrying at least two pinholes, said several plates being placed against each other and being able to slide with respect to each other, in order to form the modifiable set of at least one pinhole by the superimposition of pinholes (114, 101) in said several plates, and to modify, by sliding said plates with respect to each other, the number and/or size of the pinholes in said modifiable set of at least one pinhole.","2.A modifiable set of at least one pinhole as claimed in claim 1, wherein said two plates (100, 110) are transparent windows on which said pinholes are produced by the deposition of an opaque layer by a lithographic method, and wherein said opaque layers of these two plates are turned towards each other, so that the space between said opaque layers is as small as possible.","3.A modifiable set of at least one pinhole as claimed in one of claims 1 or 2, wherein at least one of said plates (1040; 1041) is a fine opaque sheet in which said pinholes are obtained by piercing.","4.A modifiable set as claimed in claim 3, wherein said plates (1040, 1041, 1042, 1051) consisting of fine opaque sheets are placed against each other and clamped between two thick plates, in order to prevent any deformation of said plates consisting of fine opaque sheets.","5.A modifiable set as claimed in one of claims 1 to 4, wherein said plates are separated from each other by layers of a transparent lubricating liquid.","6.A modifiable set as claimed in one of claims 1 to 5, wherein the sliding of one of said plates with respect to an adjacent plate takes place by translation along an axis, and wherein these two plates are positioned with respect to each other by means of microscopic guide rails (1101, 1102,1103, 1111, 1112, 1113).","7.A modifiable set of pinholes as claimed in claim 6, comprising several microscopic guide rails.","8.A modifiable set of pinholes as claimed in claim 7, wherein the microscopic guide rails are produced by lithography.","9.A modifiable set of pinholes as claimed in one of claims 1 to 8, wherein: it comprises a first intermediate set of pinholes (101) formed by one or more of said plates (100), and a second intermediate set of pinholes (115, 114, 113, 112, 111) formed by one or more of said plates (110), it comprises a means for making the second intermediate set slide with respect to the first intermediate set so as to pass from a first configuration to a second configuration, at least one hole in the second intermediate set which is superimposed on a hole in the first intermediate set in the first configuration is not superimposed on a hole in the first intermediate set in the second configuration, so that the number and/or size of the holes is determined by the number and/or size of the holes in the second intermediate set which are superimposed on the holes in the first intermediate set.","10.A modifiable set of pinholes as claimed in claim 9, wherein the number of holes in the second intermediate set (320) which are superimposed on holes in the first intermediate set (310) in the first configuration differs from the number of holes in the second intermediate set which are superimposed on holes in the first intermediate set in the second configuration, so that the number of holes in the modifiable set of pinholes in the second configuration differs from the number of holes in the modifiable set of pinholes in the first configuration.","11.A modifiable set of pinholes as claimed in claim 9, wherein the size of the holes in the second intermediate set (300) which are superimposed on holes in the first intermediate set (310) in the first configuration differs from the size of the holes in the second intermediate set which are superimposed on holes in the first intermediate set in the second configuration, so that the size of the holes in the modifiable set of pinholes in the second configuration differs from the size of the holes in the modifiable set of pinholes in the first configuration.","12.A modifiable set of pinholes as claimed in one of claims 1 to 8, comprising means for continuously moving the plates (400) with respect to each other so that this continuous movement results in a continuous modification of the surface of each pinhole.","13.A modifiable set of pinholes as claimed in claim 12, wherein, in a reference position, the pinholes in said several plates (400) are exactly superimposed on each other.","14.A modifiable set of pinholes as claimed in one of claims 12 or 13, comprising means for generating a translation movement of the ith plate in a direction oriented at 2 π N i radians in a reference frame common to the N plates constituting the set of pinholes.","15.A modifiable set of pinholes as claimed in one of claims 12 to 14, wherein the pinholes are polygons with 2N sides, where N is the number of plates constituting the set of pinholes.","16.A modifiable set of pinholes as claimed in claims 14 and 15, wherein the directions of the translation movement of the plates with respect to each other are directed along midperpendiculars of the polygons.","17.A modifiable set of pinholes as claimed in one of claims 12 to 16, wherein the sliding of the plates with respect to each other is obtained by means of an iris diaphragm mechanism (FIG.","16).","18.A modifiable set of pinholes as claimed in one of claims 1 to 16, wherein one of the plates (110) is moved by means of a linear positioner along an axis.","19.A modifiable set of pinholes as claimed in one of claims 1 to 16, wherein one of the plates (1051) is moved by means of a two-axis positioner.","20.A modifiable set of pinholes as claimed in one of claims 1 to 19, comprising exactly two plates.","21.A modifiable set of pinholes as claimed in one of claims 1 to 20, comprising at least two pinholes."],["<SOH> TECHNICAL FIELD <EOH>The invention concerns a microscopic hole or a set of microscopic holes (pinholes), the number of these holes and/or their size being able to be modified easily.","Such a set of pinholes is intended to be used for various applications in optics, in particular in confocal microscopy."],["TECHNICAL FIELD The invention concerns a microscopic hole or a set of microscopic holes (pinholes), the number of these holes and/or their size being able to be modified easily.","Such a set of pinholes is intended to be used for various applications in optics, in particular in confocal microscopy.","STATE OF THE ART In confocal microscopy use is usually made of two types of pinhole: holes of fixed size: to modify the size of a hole, it is necessary to replace it with another.","Typically several pinholes can be mounted on a wheel having a position corresponding to the use of each of these holes.","The movement of the wheel must be very precise.","holes of variable size: functioning on the principle of the iris diaphragm, they require at least three blades which form a hole by crossing one another and are expensive because of the relative complexity of the mechanism.","Conventionally, confocal microscopy systems require the use of a single pinhole.","For example, the first embodiment of French patent application number 0103860 of Mar.","22, 2001, as well as the microscope described in FIG.","3 of the U.S. Pat.","No.","5,978,095 or the microscope described in the patent application U.S. Pat.","No.","5,162,941.Other confocal microscopy systems require the use of an array of pinholes.","For example, the microscopes described by FIG.","1 of U.S. Pat.","No.","5,239,178 or FIG.","3 in the patent U.S. Pat.","No.","5,978,095, or the Nipkow disk systems.","In certain embodiments of a microscope such as the one described in French patent number 0103860 of Mar.","22, 2001, an array of pinholes must be positioned with great precision, which is difficult using a simple technique consisting of exchanging the whole of the array.","When “single” pinholes are simply exchanged, as on certain single-point confocal microscopes, their precise positioning is also difficult.","In addition, the systems for exchanging arrays of pinholes are necessarily bulky, since their size is the sum of the sizes of each array able to be exchanged.","In the case of microscopes using an array of pinholes, the size and density of the holes cannot usually be modified.","However, this modification is desirable in order to adapt the size of the holes to the wavelength being studied.","U.S. Pat.","No.","6,002,509 affords a solution to this problem in the case of a Nipkow disk microscope.","However, this solution requires the replacement of the array of holes with an array of reflective points.","When the technique used consists of using reflective points produced by a multilayer treatment, each wavelength corresponds to a given size and density of the reflective points.","It is then not possible to modify the size or density of the holes of the hole with a given wavelength, and the number of different sizes of holes is limited by the performance of the multilayer treatment.","When the technique used consists of introducing several concentric rings on the Nipkow disk, a movement of the disk, which is not very practical, is necessary, and the size of the disk rapidly becomes excessive.","The technique is difficult to adapt to systems using a fixed array of pinholes.","DESCRIPTION OF THE INVENTION The object of the invention is a set of one or more pinholes of variable size and/or number, the changes to which are obtained by a simplified method which is precise and inexpensive.","In particular, one object of the invention is to produce pinholes which can be modified without problems in positioning and which are of reduced bulk.","“Holes” means holes in the optical sense of the term, that is to say small areas through which light can pass, not necessarily void.","A “hole” can for example be an interruption in an opaque layer deposited on glass.","To this end, the object of the invention is a modifiable set of at least one pinhole intended for filtering a light beam, comprising several plates each carrying at least one pinhole, one of these plates carrying at least two pinholes, said several plates being placed against each other and being able to slide with respect to each other, in order to form the modifiable set of at least one pinhole by the superimposition of pinholes in said several plates, and to modify, by sliding said plates with respect to each other, the number and/or size of the pinholes in said modifiable set of at least one pinhole.","An iris diaphragm also comprises plates sliding with respect to each other.","The invention is distinguished in this by the fact that these plates carry pinholes and by the fact that one of the plates carries at least two pinholes.","This particular arrangement simplifies the design of the system (in the case of a single pinhole, it is possible to use only two plates) and makes it possible to produce arrays of modifiable pinholes, whilst iris diaphragms are designed only for a single modifiable pinhole.","A modifiable set of pinholes can also be obtained by a system physically exchanging two sets of pinholes produced on different plates.","This solution is used in certain single-point confocal microscopes.","The present invention is distinguished from this simple technical solution by the use of several superimposed plates, which makes it possible to modify the array of pinholes by means of movements which are also microscopic, rather than macroscopic as is the case in the state of the art.","This simplifies the positioning problems.","Various techniques for producing plates can be employed.","For example, and according to one characteristic of the invention, two of said plates can be transparent windows on which said pinholes are produced by the deposition of an opaque layer by a lithographic method.","The opaque layers on these two plates can then be turned towards each other, so that the space separating them is as small as possible.","The advantage of this technique is that the windows have good rigidity (deform little).","According to one characteristic of the invention, at least one of the plates is a fine opaque sheet in which said pinholes are obtained by piercing.","This solution is necessary when more than two plates are used.","This is because, if only glass plates are used, their thickness does not make it possible to produce an array of holes correctly.","According to one characteristic of the invention, the plates consisting of fine opaque sheets are placed against each other and held between two thick plates, in order to prevent any deformation of said plates consisting of fine opaque sheets.","This is because one difficulty in production is the tendency to the deformation of the fine sheets, which do not have the necessary rigidity and must therefore be placed between thicker supports.","According to one characteristic of the invention, the plates are separated from each other by layers of a transparent lubricating liquid.","This is because, in the contrary case, friction between the plates make correct functioning difficult.","Another solution is to use plates which do not touch each other, but this solution is difficult since it requires excellent surface evenness of the plates.","Sliding of one plate with respect to another can in general take place along two axes.","However, the system is simplified, according to one characteristic of the invention, if this sliding takes place along only one axis.","In this case, it is possible to use a guide rail to help maintain correct relative positioning of the plates.","However, such a rail is expensive and poses problems of positioning.","In order to facilitate the relative positioning of the plates, and according to one characteristic of the invention, two adjacent plates sliding with respect to each other along one axis are positioned with respect to each other by microscopic guide rails.","A microscopic rail being fragile, it is preferable, according to one characteristic of the invention, to use several microscopic guide rails.","These rails can for example be produced by lithography.","Various solutions can be used for the arrangement of the plates and the distribution of the holes on the plates.","According to one characteristic of the invention, the modifiable set of pinholes satisfies the following facts: it comprises a first intermediate set of pinholes formed by one or more of said plates, and a second intermediate set of pinholes formed by one or more of said plates, it comprises a means for making the second intermediate set slide with respect to the first intermediate set so as to pass from a first configuration to a second configuration, at least one hole in the second intermediate set which is superimposed on a hole in the first intermediate set in the first configuration is not superimposed on a hole in the first intermediate set in the second configuration, so that the number and/or size of the holes is determined by the number and/or size of the holes in the second intermediate set which are superimposed on the holes in the first intermediate set.","In this case, the change in configuration is accompanied by a change in the pinhole or pinholes in the second intermediate set which is used.","The change in configuration therefore consists of making the plates slide with respect to each other so as to bring a new pinhole in the second intermediate set opposite a pinhole in the first intermediate set.","Typically, a discrete number of configurations is used, and the relative movement of the plates takes place step by step.","According to one characteristic of the invention, the number of holes in the second intermediate set which are superimposed on holes in the first intermediate set in the first configuration differs from the number of holes in the second intermediate set which are superimposed on holes in the first intermediate set in the second configuration, so that the number of holes in the modifiable set of pinholes in the second configuration differs from the number of holes in the modifiable set of pinholes in the first configuration.","According to one characteristic of the invention, the size of the holes in the second intermediate set which are superimposed on holes in the first intermediate set in the first configuration differs from the size of the holes in the second intermediate set which are superimposed on holes in the first intermediate set in the second configuration, so that the size of the holes in the modifiable set of pinholes in the second configuration differs from the size of the holes in the modifiable set of pinholes in the first configuration.","These two characteristics of the invention can be combined in order to simultaneously modify the size and number of pinholes.","For example, it is possible to use only two plates each constituting an intermediate set, and to vary the size and/or number of holes solely by moving one of these plates along one axis.","This solution is effective and simple.","However, it is insufficient when it is sought to generate an array of pinholes able to have a high density of pinholes.","This is because, in this case, sufficient space is not available on the plate constituting the first intermediate set to conceal the various pinholes in the second intermediate set which must successively be superimposed on the same hole in the first intermediate set.","According to one characteristic of the invention, an appropriate solution to this problem consists of continuously moving the plates with respect to each other so that this continuous movement results in a continuous modification of the surface of each pinhole in the modifiable set.","It is possible for example to use several identical plates and one reference position for which the pinholes in said several plates are exactly superimposed on each other.","So that the reduction in size of the pinholes takes place regularly, it is preferable to generate a translation movement of the ith plate in a direction oriented at 2 π N radians in a reference frame common to the N plates constituting the set of pinholes.","In the case where N plates are used, the pinholes are preferably polygons with 2N sides, although they can also have other shapes, for example circular.","The directions of the translation movement of the plates with respect to each other are then preferably directed along midperpendiculars of the polygons.","According to one characteristic of the invention, the sliding of the plates with respect to each other is obtained by means of an iris diaphragm mechanism.","This solution is well adapted to the previous case, where it makes it possible to coordinate the continuous movement of several plates.","This iris diaphragm mechanism can, in the case where a rotation of the set of pinholes must be avoided, be supplemented by a supplementary rotation device for compensating for the rotation caused by an iris diaphragm mechanism with a single movable element.","According to one characteristic of the invention, one of the plates is moved by means of a linear positioner along an axis.","This solution is preferred when a technique based on discrete movements is used.","It may also be necessary to move one of the plates by means of a two-axis positioner.","This solution is one which allows the maximum flexibility.","When the density of pinholes sought is low, and according to one characteristic of the invention, the modifiable set of pinholes can easily be produced by means of two plates only.","In this case it may be a case of thick transparent plates on which the arrays of holes are produced by a lithographic method.","This solution is the most simple, in particular because the plates do not deform.","However, when the density of pinholes is high, it becomes necessary to use more than two plates, as seen above.","The modifiable set of pinholes according to the invention may comprise only one pinhole of modifiable size.","It is then useful in a single-point confocal microscope.","It may, according to one characteristic of the invention, comprise several pinholes of modifiable size, number and/or location, in which case it is useful with a confocal microscope with multipoint illumination.","QUICK DESCRIPTION OF THE FIGURES FIG.","1 depicts a fixed plate used in a first embodiment.","FIG.","2 depicts a movable plate used in this embodiment.","FIG.","3 depicts the superimposition of the two plates, in this embodiment, forming a single pinhole of small size.","FIG.","4 depicts the same superimposition in section.","FIG.","5 depicts a movable plate used in a second embodiment.","FIG.","6 depicts a fixed plate used in this embodiment.","FIG.","7 depicts a movable plate used in a third embodiment.","FIG.","8 depicts a fixed plate used in this third embodiment.","FIG.","9 depicts a movable plate used in a variant of this third embodiment.","FIG.","10 depicts a plate used in a fourth embodiment.","FIG.","11 depicts a device used for preventing leakages of the liquid separating the plates, the device being filled under vacuum.","FIG.","12 depicts a similar device but comprising an overflow having a breather.","FIG.","13 depicts a plate used in a fifth embodiment.","FIG.","14 illustrates the movement of 3 plates in this embodiment.","FIG.","15 shows a supplementary plate used in this embodiment.","FIG.","16 shows in front view the device driving 3 identical plates on the principle in FIG.","14.FIG.","17 shows this device in section, supplemented by a fourth plate.","FIGS.","18 to 21 relate to a method of guiding the plates by means of microscopic rails.","FIG.","18 depicts a movable plate carrying rails.","FIG.","19 depicts a movable plate carrying rails corresponding to those of FIG.","18.FIG.","20 depicts in section the assembly of two plates produced on plates of thick glass.","FIG.","21 depicts in section the assembly of two plates produced on metallic sheets.","FIRST EMBODIMENT This first embodiment makes it possible to obtain, by step by step sliding, a movable plate 110 constituting a second intermediate set, with respect to a fixed plate 100 constituting a first intermediate set, a single pinhole of modifiable size in the form of a disk.","FIG.","1 depicts the plate 100 comprising a hole 101.FIG.","2 depicts the plate 110 comprising holes 111, 112, 113, 114, 115 aligned on an axis 116.The modifiable pinhole is formed by the two plates A and B placed one against the other.","FIG.","3 depicts the modifiable pinhole obtained when the hole 114 is superimposed on the hole 101.FIG.","4 depicts the same superimposition in section along a plane passing through the axis 116.In the position of the plates which is depicted in FIGS.","3 and 4, only the hole 114 can have light passing through it, and the diameter of the modifiable hole is that of the hole 114.The plate 110 must be mounted on a positioner making it possible to move it in the direction of the axis 116 passing through the set of holes 111 to 115.By sliding the plate 110 with respect to the plate 100, by means of the positioner, it is possible to superimpose any one of the holes 111 to 115 on the hole 101 and therefore to obtain five distinct diameters of the modifiable pinhole.","When for example the hole 113, instead of the hole 114, is superimposed on the hole 101, the diameter of the modifiable hole is increased and becomes equal to the diameter of the hole 113.The pinholes in the plate 110 are not necessarily aligned on the same straight line, however this solution makes it possible to move the plate 110 only along one axis, which minimizes costs.","In general terms, it is possible to obtain as many holes with different characteristics as there are holes formed in the plate 110.A guide rail can be used so that the movement of the plate B with respect to the plate A is restricted to the direction of the axis 116.The plates 100 and 110 can be glass plates whose opaque parts are metallic layers obtained by a lithographic process.","These metallic layers are shown in thick lines in FIG.","4.The two plates are separated by a fine layer 117 of optical liquid, for example oil, in order to prevent any solid rubbing.","When the optical liquid is maintained solely by capillary attraction, as indicated in FIG.","4, the qualities of the layer are liable to degrade over time by partial evaporation, and the liquid is also liable to spread into the parts of the optical system other than the areas where its presence is required.","In order to avoid these problems the system can be made impermeable as indicated in FIG.","14 by means of a flexible closure 500, for example made from plastic, which closes the whole of the system.","The liquid can then be injected under vacuum between the two plates and into the area included inside the flexible closure 500.This device makes it possible to reconcile the movement of the plates 110 and 100 with the absence of leakages of liquid.","One alternative to filling under vacuum is the overflow system depicted in FIG.","15.A tube 105 leads into a reservoir 502 provided with a breather and raised up and ensures the maintenance of a level of optical liquid in the area included between the plates.","In all the embodiments use will preferably be made of glass plates with opaque layers obtained by lithography and turned towards each other, separated by an optical liquid and equipped with a system preventing leakages of liquid.","These technological aspects will not be repeated in the remainder of the explanations.","SECOND EMBODIMENT This second embodiment makes it possible to obtain an array of pinholes of modifiable size by step by step sliding of a movable plate 300 on a fixed plate 310.FIG.","5 depicts an example of a plate 300 and FIG.","6 depicts the corresponding plate 310.The array of modifiable pinholes is formed by plates 310 and 300 placed against each other.","When the hole 301 in the plate 300 is superimposed on the hole 311 in the plate 310, the orientation of the plates not being modified compared with the drawing, the array of modifiable pinholes has the appearance of the plate 310 alone, that is to say all the holes in the plate 310 are left free by the plate 300.When the hole 302 in the plate 300 is superimposed on the hole 311 in the plate 310, the diameter of the holes in the array of modifiable pinholes is decreased.","When the holes 303, 304, 305 are successively superimposed on the hole 311 in the plate 310, the diameter of the holes in the array of modifiable pinholes is on each occasion decreased.","There is therefore in this example an array of modifiable pinholes whose diameter can take 5 distinct values.","In general terms, it is possible to produce on this principle an array comprising a large number of pinholes each able to take various sizes or shapes.","THIRD EMBODIMENT This third embodiment makes it possible to obtain holes of variable size and number by sliding a movable plate 320 carrying a second intermediate set of holes on a fixed plate 330 carrying a first intermediate set of holes.","FIG.","7 depicts an example of a plate 320 used and FIG.","8 depicts the corresponding plate 330.When the hole 321 in the plate 320 is superimposed on the hole 331 in the plate 330, the array of modifiable pinholes has the appearance of the plate 330 alone, that is to say the 12 holes in the plate 330 are left free by the plate 320.When for example the hole 322 in the plate 320 is superimposed on the hole 331 in the plate 330, the diameter of the holes in the array of modifiable pinholes is decreased but their number is constant.","When the hole 323 in the plate 320 is superimposed on the hole 331 in the plate 330, the number of pinholes in the set of modifiable pinholes becomes equal to 4 instead of 12, their diameter being equal to that of the holes in the plate 330, that is to say 6 holes in the plate 330 are concealed by the opaque part of the plate 320, and the other 4 holes are left free.","When the hole 324 in the plate 320 is superimposed on the hole 331 in the plate 330, the modifiable set of pinholes comprises 4 holes of reduced diameter.","This example therefore constitutes a modifiable set of pinholes, the number of pinholes being able to be equal to 4 or 12, and the diameter of each hole being able to take 5 distinct values.","In general terms it is possible to produce on this principle a modifiable array comprising a large number of holes, the size and number of which are both modifiable.","Another example of this embodiment uses the same plate 330 but the second intermediate set is formed by the plate 340 depicted in FIG.","9.When the hole 341 in the plate 340 is superimposed on the hole 331 in the plate 330, the array of pinholes modifiable to the appearance of the plate 330 alone, that is to say the 12 holes in the plate 330, are left free by the plate 340.When for example the hole 342 in the plate 340 is superimposed on the hole 331 in the plate 330, the diameter of the holes in the array of modifiable pinholes is decreased but their number is constant.","When the hole 343 in the plate 340 is superimposed on the hole 332 in the plate 330, the number of pinholes in the set of modifiable pinholes becomes equal to 1 instead of 12, their diameter being equal to that of the holes in the plate 330, that is to say 11 holes in the plate 330 are concealed by the opaque part of the plate 340 and the other 1 hole is left free.","When the hole 344 in the plate 340 is superimposed on the hole 332 in the plate 330, the modifiable set of pinholes comprises only 1 hole of reduced diameter.","This example therefore constitutes a modifiable set of pinholes, the number of pinholes being able to be equal to 1 or 12, and the diameter of each hole being able to take 5 distinct values.","This type of array of pinholes is particularly useful in a confocal microscope which it is wished to be able to use both in multipoint mode and in single-point mode.","FOURTH EMBODIMENT This fifth embodiment makes it possible to obtain square holes of continuously variable size.","It uses two identical plates depicted for example by FIG.","10.When the hole 401 in the first plate is exactly superimposed on the hole 401 in the second plate a set of square holes equivalent to the first plate alone is obtained.","When the two plates are moved with respect to each other along the axis 402, the size of the square holes resulting from the superimposition of the plates is decreased.","FIFTH EMBODIMENT This fifth embodiment is particularly adapted to the case where a high density of pinholes is sought.","In a basic version, it requires the use of 3 plates which move continuously with respect to each other.","They are driven by means of a diaphragm device with modified iris in order to compensate for the rotation of the whole.","The pinholes are hexagonal.","The three plates carry arrays of identical holes which, in a reference position, are superimposed on each other.","FIG.","13 shows a plate 1000 comprising pinholes, for example 1001.The broken lines, for example 1002, delimit hexagonal locations not carrying any hole.","In the reference position, the appearance of the modifiable set of pinholes is the same as the appearance of each of the plates and is therefore depicted by FIG.","13.The size of the holes in the modifiable set of pinholes is modified by moving the plates with respect to each other in the manner indicated by FIG.","14.This figure depicts part of the array of pinholes.","In the figure, the broken lines depict the limits of the pinholes in each of the three plates and the intersection of these holes, which constitutes the effective hole of the modifiable array, has been depicted in white.","The arrows show the direction of movement of the plates from the reference position.","By moving the plates in the direction of the arrows the width of the holes is decreased continuously.","The shape of the holes of the modifiable set of pinholes is not modified when their width decreases.","This is due to the fact that there are 3 plates, that the holes have 2×3=6 sides, and that the directions of the movement are along the midperpendiculars of the hexagons.","In a version also making it possible to modify the density of pinholes, a fourth plate is necessary.","This plate is depicted in FIG.","15.The set of modifiable pinholes obtained by means of three plates and depicted in FIG.","13 in the reference position constitutes a first intermediate set.","The plate in FIG.","14 constitutes a second intermediate set.","When the hole 1010 in the plate in FIG.","15 is superimposed on the hole 1001 in the first intermediate set in FIG.","13, the density of pinholes is at a maximum.","When the hole 1013 in the plate in FIG.","15 is superimposed on the hole 1001 in the first intermediate set in FIG.","13, the number of pinholes per unit surface area is divided by 4.When the hole 1012 in the plate in FIG.","15 is superimposed on the hole 1001 in the first intermediate set in FIG.","13, the number of pinholes per unit surface area is divided by 9.When the hole 1011 in the plate in FIG.","15 is superimposed on the hole 1001 in the first intermediate set in FIG.","13, the number of pinholes per unit surface area is divided by 16.The three identical plates depicted in FIG.","13 can be driven by means of an iris diaphragm drive system depicted in FIGS.","16 and 17.The plates in FIG.","13 are the plates 1040, 1041, 1042 depicted in FIG.","17.These plates are metallic sheets tensioned over circular holding rings 1022, 1021, 1020 and carrying holes produced for example by laser piercing.","These three fixing rings are connected to two control rings 1023, 1024.For example, the internal ring 1020 is connected to the control ring 1023 by a bar 1028 turning freely about an axis 1029 fixed in the control ring 1023.The internal ring 1020 is connected to the control ring 1024 by a bar 1026 turning freely about an axis 1027 fixed in the control ring 1024.The housing 1031 of the axis 1027 is oversized so as to be able to combine a rotation and translation with respect to the axis 1027.A tie rod 1030 is used for keeping the housing 1031 in abutment on the axis 1027.The other two holding rings are connected in a similar manner to the control rings.","When the two control rings turn simultaneously in opposite directions and by an equal angle, the three plates move in translation with an angle of 120 degrees between the directions of each movement axis, as indicated in FIG.","14.FIG.","17 also depicts the fourth plate 1051 which is a sheet tensioned on a holding ring 1050.The holding ring 1050 is mounted on a two-axis positioner.","The plates 1051 and 1042 are themselves pressed on thick plates of glass 1052 and 1043.When the whole of the system is in position the two glass plates 1052 and 1043 prevent deformations of the sheets (plates) 1042, 1041, 1040, 1051 carrying pinholes.","Method of Guiding and Positioning the Plates In certain embodiments the plates carrying pinholes move in translation with respect to each other along a single axis.","For example, in the first embodiment, the second and fourth embodiments, and also in the fifth embodiment with regard to the three plates moving by means of an iris diaphragm mechanism.","This solution simplifies the system in that each plate moves with respect to another along a single axis.","As indicated above a guide rail can be used for guiding the plates.","However, a macroscopic guide rail is difficult to produce with the required precision.","In order to obtain good positioning of the plates it is possible to replace such a guide rail with a set of microscopic guide rails.","FIGS.","18 to 20 illustrate the embodiment of a set of microscopic guide rails in a system with two plates of the type described in the first embodiment.","FIG.","18 shows the female part 1101, 1102, 1103 of the rails, produced by lithography in the same way as the pinholes, on the movable plate.","FIG.","19 shows the male part 1111, 1112, 1113 of the rails, produced by lithography on the fixed plate, but by means of a supplementary metallic layer.","FIG.","20 shows in section the assembly of the rails 1103 and 1113.The cross-section is produced along a cutting axis 1114 of the fixed plate and along a cutting axis 1104 of the movable plate, these two axes being deemed to be superimposed in the position depicted.","The fixed glass plate 1120 carries a metallic layer 1122 leaving the pinhole 1115 free.","It also carries a supplementary metallic layer 1113 constituting the male part of the guidance system.","This male part is produced by lithography and the remains of a layer of resin 1125 have also been shown.","The object of this layer of resin is to protect the bottom metallic layer 1122 when the rail 1113 is formed.","The movable plate 1120 carries a metallic layer 1123 leaving free the pinhole 1105 and the female part 1103 of the guide rail.","The space 1123 included between the two plates is filled with a lubricating liquid for preventing solid friction.","The male part 1113 fits in the groove 1103 in order to guide the movement.","The superimposition of the pinholes 1115 and 1105 produces a pinhole of reduced size.","This guidance method can be adapted to the case where the plates are metallic sheets, as in the fifth embodiment.","In this case male or female rails can be produced on each side of each sheet.","The glass plates are then replaced by the metallic sheets.","The diagram in FIG.","21 is equivalent to that in FIG.","20 but illustrates the case of plates formed by fine metallic sheets.","The plate 1134 carries the male rail 1132 separated from the plate by a layer of protective resin 1133 used for producing the rail 1132 by lithography.","The plate 1137 carries the female rail 1138 produced by lithography in a metallic layer 1135 separated from the plate by a resin 1139.The space between the two plates is filled with a lubricating liquid.","A guide rail has been shown only on one side of each plate but it is possible to produce one on each side of each plate.","In the case of the fifth embodiment, the guide rails which separate each metallic sheet guided by two adjacent sheets participate in the maintenance of the shape of the metallic sheets and in the prevention of deformations.","Naturally sheet number 1051 in FIG.","17 can be guided by this method only if its movement is restricted to a single direction.","Industrial Applications The present set of pinholes can be used in a confocal microscope with single-point or multipoint illumination or in a confocal microscope intended to alternate the two illumination modes.","For example, if a set of pinholes of the type described in the fourth embodiment replaces the set of pinholes used in the system described by FIG.","1 of U.S. Pat.","No.","5,239,178 it becomes possible to modify the size and number of these pinholes and possibly to alternate between a multipoint and a single-point operating mode.","Likewise, the array of pinholes in the fourth embodiment of the present invention can replace, with the same effect, the array of pinholes used in FIG.","3 of the U.S. Pat.","No.","5,978,095.By using a modifiable array of pinholes according to the present invention in the microscope described by one of the first two embodiments of French patent application number 0103860 of Mar.","22, 2001, it is possible to easily modify the diameter of the pinholes or their number, which affects the speed/resolution or speed/penetration depth compromise in the sample.","The pinhole described in one of the first two embodiments of the present invention can also replace the interchangeable pinholes or the “iris” diaphragms normally used in single-point confocal microscopes with laser scanning."]],"string":"[\n [\n \"Modifiable assembly of microscopic apertures\",\n \"The invention concerns a modifiable assembly of microscopic apertures comprising several plates (100, 110) coated with an opaque deposition except on transparent parts, for example (101, 114, 115), capable of sliding relative to one another, separated by an oil layer (117).\",\n \"The invention is applicable to microscopic apertures for single-point or multiple-point confocal microscopy.\"\n ],\n [\n \"1.A modifiable set of at least one pinhole intended for filtering a light beam, comprising several plates (100, 110) each carrying at least one pinhole, one of these plates (110) carrying at least two pinholes, said several plates being placed against each other and being able to slide with respect to each other, in order to form the modifiable set of at least one pinhole by the superimposition of pinholes (114, 101) in said several plates, and to modify, by sliding said plates with respect to each other, the number and/or size of the pinholes in said modifiable set of at least one pinhole.\",\n \"2.A modifiable set of at least one pinhole as claimed in claim 1, wherein said two plates (100, 110) are transparent windows on which said pinholes are produced by the deposition of an opaque layer by a lithographic method, and wherein said opaque layers of these two plates are turned towards each other, so that the space between said opaque layers is as small as possible.\",\n \"3.A modifiable set of at least one pinhole as claimed in one of claims 1 or 2, wherein at least one of said plates (1040; 1041) is a fine opaque sheet in which said pinholes are obtained by piercing.\",\n \"4.A modifiable set as claimed in claim 3, wherein said plates (1040, 1041, 1042, 1051) consisting of fine opaque sheets are placed against each other and clamped between two thick plates, in order to prevent any deformation of said plates consisting of fine opaque sheets.\",\n \"5.A modifiable set as claimed in one of claims 1 to 4, wherein said plates are separated from each other by layers of a transparent lubricating liquid.\",\n \"6.A modifiable set as claimed in one of claims 1 to 5, wherein the sliding of one of said plates with respect to an adjacent plate takes place by translation along an axis, and wherein these two plates are positioned with respect to each other by means of microscopic guide rails (1101, 1102,1103, 1111, 1112, 1113).\",\n \"7.A modifiable set of pinholes as claimed in claim 6, comprising several microscopic guide rails.\",\n \"8.A modifiable set of pinholes as claimed in claim 7, wherein the microscopic guide rails are produced by lithography.\",\n \"9.A modifiable set of pinholes as claimed in one of claims 1 to 8, wherein: it comprises a first intermediate set of pinholes (101) formed by one or more of said plates (100), and a second intermediate set of pinholes (115, 114, 113, 112, 111) formed by one or more of said plates (110), it comprises a means for making the second intermediate set slide with respect to the first intermediate set so as to pass from a first configuration to a second configuration, at least one hole in the second intermediate set which is superimposed on a hole in the first intermediate set in the first configuration is not superimposed on a hole in the first intermediate set in the second configuration, so that the number and/or size of the holes is determined by the number and/or size of the holes in the second intermediate set which are superimposed on the holes in the first intermediate set.\",\n \"10.A modifiable set of pinholes as claimed in claim 9, wherein the number of holes in the second intermediate set (320) which are superimposed on holes in the first intermediate set (310) in the first configuration differs from the number of holes in the second intermediate set which are superimposed on holes in the first intermediate set in the second configuration, so that the number of holes in the modifiable set of pinholes in the second configuration differs from the number of holes in the modifiable set of pinholes in the first configuration.\",\n \"11.A modifiable set of pinholes as claimed in claim 9, wherein the size of the holes in the second intermediate set (300) which are superimposed on holes in the first intermediate set (310) in the first configuration differs from the size of the holes in the second intermediate set which are superimposed on holes in the first intermediate set in the second configuration, so that the size of the holes in the modifiable set of pinholes in the second configuration differs from the size of the holes in the modifiable set of pinholes in the first configuration.\",\n \"12.A modifiable set of pinholes as claimed in one of claims 1 to 8, comprising means for continuously moving the plates (400) with respect to each other so that this continuous movement results in a continuous modification of the surface of each pinhole.\",\n \"13.A modifiable set of pinholes as claimed in claim 12, wherein, in a reference position, the pinholes in said several plates (400) are exactly superimposed on each other.\",\n \"14.A modifiable set of pinholes as claimed in one of claims 12 or 13, comprising means for generating a translation movement of the ith plate in a direction oriented at 2 π N i radians in a reference frame common to the N plates constituting the set of pinholes.\",\n \"15.A modifiable set of pinholes as claimed in one of claims 12 to 14, wherein the pinholes are polygons with 2N sides, where N is the number of plates constituting the set of pinholes.\",\n \"16.A modifiable set of pinholes as claimed in claims 14 and 15, wherein the directions of the translation movement of the plates with respect to each other are directed along midperpendiculars of the polygons.\",\n \"17.A modifiable set of pinholes as claimed in one of claims 12 to 16, wherein the sliding of the plates with respect to each other is obtained by means of an iris diaphragm mechanism (FIG.\",\n \"16).\",\n \"18.A modifiable set of pinholes as claimed in one of claims 1 to 16, wherein one of the plates (110) is moved by means of a linear positioner along an axis.\",\n \"19.A modifiable set of pinholes as claimed in one of claims 1 to 16, wherein one of the plates (1051) is moved by means of a two-axis positioner.\",\n \"20.A modifiable set of pinholes as claimed in one of claims 1 to 19, comprising exactly two plates.\",\n \"21.A modifiable set of pinholes as claimed in one of claims 1 to 20, comprising at least two pinholes.\"\n ],\n [\n \"<SOH> TECHNICAL FIELD <EOH>The invention concerns a microscopic hole or a set of microscopic holes (pinholes), the number of these holes and/or their size being able to be modified easily.\",\n \"Such a set of pinholes is intended to be used for various applications in optics, in particular in confocal microscopy.\"\n ],\n [\n \"TECHNICAL FIELD The invention concerns a microscopic hole or a set of microscopic holes (pinholes), the number of these holes and/or their size being able to be modified easily.\",\n \"Such a set of pinholes is intended to be used for various applications in optics, in particular in confocal microscopy.\",\n \"STATE OF THE ART In confocal microscopy use is usually made of two types of pinhole: holes of fixed size: to modify the size of a hole, it is necessary to replace it with another.\",\n \"Typically several pinholes can be mounted on a wheel having a position corresponding to the use of each of these holes.\",\n \"The movement of the wheel must be very precise.\",\n \"holes of variable size: functioning on the principle of the iris diaphragm, they require at least three blades which form a hole by crossing one another and are expensive because of the relative complexity of the mechanism.\",\n \"Conventionally, confocal microscopy systems require the use of a single pinhole.\",\n \"For example, the first embodiment of French patent application number 0103860 of Mar.\",\n \"22, 2001, as well as the microscope described in FIG.\",\n \"3 of the U.S. Pat.\",\n \"No.\",\n \"5,978,095 or the microscope described in the patent application U.S. Pat.\",\n \"No.\",\n \"5,162,941.Other confocal microscopy systems require the use of an array of pinholes.\",\n \"For example, the microscopes described by FIG.\",\n \"1 of U.S. Pat.\",\n \"No.\",\n \"5,239,178 or FIG.\",\n \"3 in the patent U.S. Pat.\",\n \"No.\",\n \"5,978,095, or the Nipkow disk systems.\",\n \"In certain embodiments of a microscope such as the one described in French patent number 0103860 of Mar.\",\n \"22, 2001, an array of pinholes must be positioned with great precision, which is difficult using a simple technique consisting of exchanging the whole of the array.\",\n \"When “single” pinholes are simply exchanged, as on certain single-point confocal microscopes, their precise positioning is also difficult.\",\n \"In addition, the systems for exchanging arrays of pinholes are necessarily bulky, since their size is the sum of the sizes of each array able to be exchanged.\",\n \"In the case of microscopes using an array of pinholes, the size and density of the holes cannot usually be modified.\",\n \"However, this modification is desirable in order to adapt the size of the holes to the wavelength being studied.\",\n \"U.S. Pat.\",\n \"No.\",\n \"6,002,509 affords a solution to this problem in the case of a Nipkow disk microscope.\",\n \"However, this solution requires the replacement of the array of holes with an array of reflective points.\",\n \"When the technique used consists of using reflective points produced by a multilayer treatment, each wavelength corresponds to a given size and density of the reflective points.\",\n \"It is then not possible to modify the size or density of the holes of the hole with a given wavelength, and the number of different sizes of holes is limited by the performance of the multilayer treatment.\",\n \"When the technique used consists of introducing several concentric rings on the Nipkow disk, a movement of the disk, which is not very practical, is necessary, and the size of the disk rapidly becomes excessive.\",\n \"The technique is difficult to adapt to systems using a fixed array of pinholes.\",\n \"DESCRIPTION OF THE INVENTION The object of the invention is a set of one or more pinholes of variable size and/or number, the changes to which are obtained by a simplified method which is precise and inexpensive.\",\n \"In particular, one object of the invention is to produce pinholes which can be modified without problems in positioning and which are of reduced bulk.\",\n \"“Holes” means holes in the optical sense of the term, that is to say small areas through which light can pass, not necessarily void.\",\n \"A “hole” can for example be an interruption in an opaque layer deposited on glass.\",\n \"To this end, the object of the invention is a modifiable set of at least one pinhole intended for filtering a light beam, comprising several plates each carrying at least one pinhole, one of these plates carrying at least two pinholes, said several plates being placed against each other and being able to slide with respect to each other, in order to form the modifiable set of at least one pinhole by the superimposition of pinholes in said several plates, and to modify, by sliding said plates with respect to each other, the number and/or size of the pinholes in said modifiable set of at least one pinhole.\",\n \"An iris diaphragm also comprises plates sliding with respect to each other.\",\n \"The invention is distinguished in this by the fact that these plates carry pinholes and by the fact that one of the plates carries at least two pinholes.\",\n \"This particular arrangement simplifies the design of the system (in the case of a single pinhole, it is possible to use only two plates) and makes it possible to produce arrays of modifiable pinholes, whilst iris diaphragms are designed only for a single modifiable pinhole.\",\n \"A modifiable set of pinholes can also be obtained by a system physically exchanging two sets of pinholes produced on different plates.\",\n \"This solution is used in certain single-point confocal microscopes.\",\n \"The present invention is distinguished from this simple technical solution by the use of several superimposed plates, which makes it possible to modify the array of pinholes by means of movements which are also microscopic, rather than macroscopic as is the case in the state of the art.\",\n \"This simplifies the positioning problems.\",\n \"Various techniques for producing plates can be employed.\",\n \"For example, and according to one characteristic of the invention, two of said plates can be transparent windows on which said pinholes are produced by the deposition of an opaque layer by a lithographic method.\",\n \"The opaque layers on these two plates can then be turned towards each other, so that the space separating them is as small as possible.\",\n \"The advantage of this technique is that the windows have good rigidity (deform little).\",\n \"According to one characteristic of the invention, at least one of the plates is a fine opaque sheet in which said pinholes are obtained by piercing.\",\n \"This solution is necessary when more than two plates are used.\",\n \"This is because, if only glass plates are used, their thickness does not make it possible to produce an array of holes correctly.\",\n \"According to one characteristic of the invention, the plates consisting of fine opaque sheets are placed against each other and held between two thick plates, in order to prevent any deformation of said plates consisting of fine opaque sheets.\",\n \"This is because one difficulty in production is the tendency to the deformation of the fine sheets, which do not have the necessary rigidity and must therefore be placed between thicker supports.\",\n \"According to one characteristic of the invention, the plates are separated from each other by layers of a transparent lubricating liquid.\",\n \"This is because, in the contrary case, friction between the plates make correct functioning difficult.\",\n \"Another solution is to use plates which do not touch each other, but this solution is difficult since it requires excellent surface evenness of the plates.\",\n \"Sliding of one plate with respect to another can in general take place along two axes.\",\n \"However, the system is simplified, according to one characteristic of the invention, if this sliding takes place along only one axis.\",\n \"In this case, it is possible to use a guide rail to help maintain correct relative positioning of the plates.\",\n \"However, such a rail is expensive and poses problems of positioning.\",\n \"In order to facilitate the relative positioning of the plates, and according to one characteristic of the invention, two adjacent plates sliding with respect to each other along one axis are positioned with respect to each other by microscopic guide rails.\",\n \"A microscopic rail being fragile, it is preferable, according to one characteristic of the invention, to use several microscopic guide rails.\",\n \"These rails can for example be produced by lithography.\",\n \"Various solutions can be used for the arrangement of the plates and the distribution of the holes on the plates.\",\n \"According to one characteristic of the invention, the modifiable set of pinholes satisfies the following facts: it comprises a first intermediate set of pinholes formed by one or more of said plates, and a second intermediate set of pinholes formed by one or more of said plates, it comprises a means for making the second intermediate set slide with respect to the first intermediate set so as to pass from a first configuration to a second configuration, at least one hole in the second intermediate set which is superimposed on a hole in the first intermediate set in the first configuration is not superimposed on a hole in the first intermediate set in the second configuration, so that the number and/or size of the holes is determined by the number and/or size of the holes in the second intermediate set which are superimposed on the holes in the first intermediate set.\",\n \"In this case, the change in configuration is accompanied by a change in the pinhole or pinholes in the second intermediate set which is used.\",\n \"The change in configuration therefore consists of making the plates slide with respect to each other so as to bring a new pinhole in the second intermediate set opposite a pinhole in the first intermediate set.\",\n \"Typically, a discrete number of configurations is used, and the relative movement of the plates takes place step by step.\",\n \"According to one characteristic of the invention, the number of holes in the second intermediate set which are superimposed on holes in the first intermediate set in the first configuration differs from the number of holes in the second intermediate set which are superimposed on holes in the first intermediate set in the second configuration, so that the number of holes in the modifiable set of pinholes in the second configuration differs from the number of holes in the modifiable set of pinholes in the first configuration.\",\n \"According to one characteristic of the invention, the size of the holes in the second intermediate set which are superimposed on holes in the first intermediate set in the first configuration differs from the size of the holes in the second intermediate set which are superimposed on holes in the first intermediate set in the second configuration, so that the size of the holes in the modifiable set of pinholes in the second configuration differs from the size of the holes in the modifiable set of pinholes in the first configuration.\",\n \"These two characteristics of the invention can be combined in order to simultaneously modify the size and number of pinholes.\",\n \"For example, it is possible to use only two plates each constituting an intermediate set, and to vary the size and/or number of holes solely by moving one of these plates along one axis.\",\n \"This solution is effective and simple.\",\n \"However, it is insufficient when it is sought to generate an array of pinholes able to have a high density of pinholes.\",\n \"This is because, in this case, sufficient space is not available on the plate constituting the first intermediate set to conceal the various pinholes in the second intermediate set which must successively be superimposed on the same hole in the first intermediate set.\",\n \"According to one characteristic of the invention, an appropriate solution to this problem consists of continuously moving the plates with respect to each other so that this continuous movement results in a continuous modification of the surface of each pinhole in the modifiable set.\",\n \"It is possible for example to use several identical plates and one reference position for which the pinholes in said several plates are exactly superimposed on each other.\",\n \"So that the reduction in size of the pinholes takes place regularly, it is preferable to generate a translation movement of the ith plate in a direction oriented at 2 π N radians in a reference frame common to the N plates constituting the set of pinholes.\",\n \"In the case where N plates are used, the pinholes are preferably polygons with 2N sides, although they can also have other shapes, for example circular.\",\n \"The directions of the translation movement of the plates with respect to each other are then preferably directed along midperpendiculars of the polygons.\",\n \"According to one characteristic of the invention, the sliding of the plates with respect to each other is obtained by means of an iris diaphragm mechanism.\",\n \"This solution is well adapted to the previous case, where it makes it possible to coordinate the continuous movement of several plates.\",\n \"This iris diaphragm mechanism can, in the case where a rotation of the set of pinholes must be avoided, be supplemented by a supplementary rotation device for compensating for the rotation caused by an iris diaphragm mechanism with a single movable element.\",\n \"According to one characteristic of the invention, one of the plates is moved by means of a linear positioner along an axis.\",\n \"This solution is preferred when a technique based on discrete movements is used.\",\n \"It may also be necessary to move one of the plates by means of a two-axis positioner.\",\n \"This solution is one which allows the maximum flexibility.\",\n \"When the density of pinholes sought is low, and according to one characteristic of the invention, the modifiable set of pinholes can easily be produced by means of two plates only.\",\n \"In this case it may be a case of thick transparent plates on which the arrays of holes are produced by a lithographic method.\",\n \"This solution is the most simple, in particular because the plates do not deform.\",\n \"However, when the density of pinholes is high, it becomes necessary to use more than two plates, as seen above.\",\n \"The modifiable set of pinholes according to the invention may comprise only one pinhole of modifiable size.\",\n \"It is then useful in a single-point confocal microscope.\",\n \"It may, according to one characteristic of the invention, comprise several pinholes of modifiable size, number and/or location, in which case it is useful with a confocal microscope with multipoint illumination.\",\n \"QUICK DESCRIPTION OF THE FIGURES FIG.\",\n \"1 depicts a fixed plate used in a first embodiment.\",\n \"FIG.\",\n \"2 depicts a movable plate used in this embodiment.\",\n \"FIG.\",\n \"3 depicts the superimposition of the two plates, in this embodiment, forming a single pinhole of small size.\",\n \"FIG.\",\n \"4 depicts the same superimposition in section.\",\n \"FIG.\",\n \"5 depicts a movable plate used in a second embodiment.\",\n \"FIG.\",\n \"6 depicts a fixed plate used in this embodiment.\",\n \"FIG.\",\n \"7 depicts a movable plate used in a third embodiment.\",\n \"FIG.\",\n \"8 depicts a fixed plate used in this third embodiment.\",\n \"FIG.\",\n \"9 depicts a movable plate used in a variant of this third embodiment.\",\n \"FIG.\",\n \"10 depicts a plate used in a fourth embodiment.\",\n \"FIG.\",\n \"11 depicts a device used for preventing leakages of the liquid separating the plates, the device being filled under vacuum.\",\n \"FIG.\",\n \"12 depicts a similar device but comprising an overflow having a breather.\",\n \"FIG.\",\n \"13 depicts a plate used in a fifth embodiment.\",\n \"FIG.\",\n \"14 illustrates the movement of 3 plates in this embodiment.\",\n \"FIG.\",\n \"15 shows a supplementary plate used in this embodiment.\",\n \"FIG.\",\n \"16 shows in front view the device driving 3 identical plates on the principle in FIG.\",\n \"14.FIG.\",\n \"17 shows this device in section, supplemented by a fourth plate.\",\n \"FIGS.\",\n \"18 to 21 relate to a method of guiding the plates by means of microscopic rails.\",\n \"FIG.\",\n \"18 depicts a movable plate carrying rails.\",\n \"FIG.\",\n \"19 depicts a movable plate carrying rails corresponding to those of FIG.\",\n \"18.FIG.\",\n \"20 depicts in section the assembly of two plates produced on plates of thick glass.\",\n \"FIG.\",\n \"21 depicts in section the assembly of two plates produced on metallic sheets.\",\n \"FIRST EMBODIMENT This first embodiment makes it possible to obtain, by step by step sliding, a movable plate 110 constituting a second intermediate set, with respect to a fixed plate 100 constituting a first intermediate set, a single pinhole of modifiable size in the form of a disk.\",\n \"FIG.\",\n \"1 depicts the plate 100 comprising a hole 101.FIG.\",\n \"2 depicts the plate 110 comprising holes 111, 112, 113, 114, 115 aligned on an axis 116.The modifiable pinhole is formed by the two plates A and B placed one against the other.\",\n \"FIG.\",\n \"3 depicts the modifiable pinhole obtained when the hole 114 is superimposed on the hole 101.FIG.\",\n \"4 depicts the same superimposition in section along a plane passing through the axis 116.In the position of the plates which is depicted in FIGS.\",\n \"3 and 4, only the hole 114 can have light passing through it, and the diameter of the modifiable hole is that of the hole 114.The plate 110 must be mounted on a positioner making it possible to move it in the direction of the axis 116 passing through the set of holes 111 to 115.By sliding the plate 110 with respect to the plate 100, by means of the positioner, it is possible to superimpose any one of the holes 111 to 115 on the hole 101 and therefore to obtain five distinct diameters of the modifiable pinhole.\",\n \"When for example the hole 113, instead of the hole 114, is superimposed on the hole 101, the diameter of the modifiable hole is increased and becomes equal to the diameter of the hole 113.The pinholes in the plate 110 are not necessarily aligned on the same straight line, however this solution makes it possible to move the plate 110 only along one axis, which minimizes costs.\",\n \"In general terms, it is possible to obtain as many holes with different characteristics as there are holes formed in the plate 110.A guide rail can be used so that the movement of the plate B with respect to the plate A is restricted to the direction of the axis 116.The plates 100 and 110 can be glass plates whose opaque parts are metallic layers obtained by a lithographic process.\",\n \"These metallic layers are shown in thick lines in FIG.\",\n \"4.The two plates are separated by a fine layer 117 of optical liquid, for example oil, in order to prevent any solid rubbing.\",\n \"When the optical liquid is maintained solely by capillary attraction, as indicated in FIG.\",\n \"4, the qualities of the layer are liable to degrade over time by partial evaporation, and the liquid is also liable to spread into the parts of the optical system other than the areas where its presence is required.\",\n \"In order to avoid these problems the system can be made impermeable as indicated in FIG.\",\n \"14 by means of a flexible closure 500, for example made from plastic, which closes the whole of the system.\",\n \"The liquid can then be injected under vacuum between the two plates and into the area included inside the flexible closure 500.This device makes it possible to reconcile the movement of the plates 110 and 100 with the absence of leakages of liquid.\",\n \"One alternative to filling under vacuum is the overflow system depicted in FIG.\",\n \"15.A tube 105 leads into a reservoir 502 provided with a breather and raised up and ensures the maintenance of a level of optical liquid in the area included between the plates.\",\n \"In all the embodiments use will preferably be made of glass plates with opaque layers obtained by lithography and turned towards each other, separated by an optical liquid and equipped with a system preventing leakages of liquid.\",\n \"These technological aspects will not be repeated in the remainder of the explanations.\",\n \"SECOND EMBODIMENT This second embodiment makes it possible to obtain an array of pinholes of modifiable size by step by step sliding of a movable plate 300 on a fixed plate 310.FIG.\",\n \"5 depicts an example of a plate 300 and FIG.\",\n \"6 depicts the corresponding plate 310.The array of modifiable pinholes is formed by plates 310 and 300 placed against each other.\",\n \"When the hole 301 in the plate 300 is superimposed on the hole 311 in the plate 310, the orientation of the plates not being modified compared with the drawing, the array of modifiable pinholes has the appearance of the plate 310 alone, that is to say all the holes in the plate 310 are left free by the plate 300.When the hole 302 in the plate 300 is superimposed on the hole 311 in the plate 310, the diameter of the holes in the array of modifiable pinholes is decreased.\",\n \"When the holes 303, 304, 305 are successively superimposed on the hole 311 in the plate 310, the diameter of the holes in the array of modifiable pinholes is on each occasion decreased.\",\n \"There is therefore in this example an array of modifiable pinholes whose diameter can take 5 distinct values.\",\n \"In general terms, it is possible to produce on this principle an array comprising a large number of pinholes each able to take various sizes or shapes.\",\n \"THIRD EMBODIMENT This third embodiment makes it possible to obtain holes of variable size and number by sliding a movable plate 320 carrying a second intermediate set of holes on a fixed plate 330 carrying a first intermediate set of holes.\",\n \"FIG.\",\n \"7 depicts an example of a plate 320 used and FIG.\",\n \"8 depicts the corresponding plate 330.When the hole 321 in the plate 320 is superimposed on the hole 331 in the plate 330, the array of modifiable pinholes has the appearance of the plate 330 alone, that is to say the 12 holes in the plate 330 are left free by the plate 320.When for example the hole 322 in the plate 320 is superimposed on the hole 331 in the plate 330, the diameter of the holes in the array of modifiable pinholes is decreased but their number is constant.\",\n \"When the hole 323 in the plate 320 is superimposed on the hole 331 in the plate 330, the number of pinholes in the set of modifiable pinholes becomes equal to 4 instead of 12, their diameter being equal to that of the holes in the plate 330, that is to say 6 holes in the plate 330 are concealed by the opaque part of the plate 320, and the other 4 holes are left free.\",\n \"When the hole 324 in the plate 320 is superimposed on the hole 331 in the plate 330, the modifiable set of pinholes comprises 4 holes of reduced diameter.\",\n \"This example therefore constitutes a modifiable set of pinholes, the number of pinholes being able to be equal to 4 or 12, and the diameter of each hole being able to take 5 distinct values.\",\n \"In general terms it is possible to produce on this principle a modifiable array comprising a large number of holes, the size and number of which are both modifiable.\",\n \"Another example of this embodiment uses the same plate 330 but the second intermediate set is formed by the plate 340 depicted in FIG.\",\n \"9.When the hole 341 in the plate 340 is superimposed on the hole 331 in the plate 330, the array of pinholes modifiable to the appearance of the plate 330 alone, that is to say the 12 holes in the plate 330, are left free by the plate 340.When for example the hole 342 in the plate 340 is superimposed on the hole 331 in the plate 330, the diameter of the holes in the array of modifiable pinholes is decreased but their number is constant.\",\n \"When the hole 343 in the plate 340 is superimposed on the hole 332 in the plate 330, the number of pinholes in the set of modifiable pinholes becomes equal to 1 instead of 12, their diameter being equal to that of the holes in the plate 330, that is to say 11 holes in the plate 330 are concealed by the opaque part of the plate 340 and the other 1 hole is left free.\",\n \"When the hole 344 in the plate 340 is superimposed on the hole 332 in the plate 330, the modifiable set of pinholes comprises only 1 hole of reduced diameter.\",\n \"This example therefore constitutes a modifiable set of pinholes, the number of pinholes being able to be equal to 1 or 12, and the diameter of each hole being able to take 5 distinct values.\",\n \"This type of array of pinholes is particularly useful in a confocal microscope which it is wished to be able to use both in multipoint mode and in single-point mode.\",\n \"FOURTH EMBODIMENT This fifth embodiment makes it possible to obtain square holes of continuously variable size.\",\n \"It uses two identical plates depicted for example by FIG.\",\n \"10.When the hole 401 in the first plate is exactly superimposed on the hole 401 in the second plate a set of square holes equivalent to the first plate alone is obtained.\",\n \"When the two plates are moved with respect to each other along the axis 402, the size of the square holes resulting from the superimposition of the plates is decreased.\",\n \"FIFTH EMBODIMENT This fifth embodiment is particularly adapted to the case where a high density of pinholes is sought.\",\n \"In a basic version, it requires the use of 3 plates which move continuously with respect to each other.\",\n \"They are driven by means of a diaphragm device with modified iris in order to compensate for the rotation of the whole.\",\n \"The pinholes are hexagonal.\",\n \"The three plates carry arrays of identical holes which, in a reference position, are superimposed on each other.\",\n \"FIG.\",\n \"13 shows a plate 1000 comprising pinholes, for example 1001.The broken lines, for example 1002, delimit hexagonal locations not carrying any hole.\",\n \"In the reference position, the appearance of the modifiable set of pinholes is the same as the appearance of each of the plates and is therefore depicted by FIG.\",\n \"13.The size of the holes in the modifiable set of pinholes is modified by moving the plates with respect to each other in the manner indicated by FIG.\",\n \"14.This figure depicts part of the array of pinholes.\",\n \"In the figure, the broken lines depict the limits of the pinholes in each of the three plates and the intersection of these holes, which constitutes the effective hole of the modifiable array, has been depicted in white.\",\n \"The arrows show the direction of movement of the plates from the reference position.\",\n \"By moving the plates in the direction of the arrows the width of the holes is decreased continuously.\",\n \"The shape of the holes of the modifiable set of pinholes is not modified when their width decreases.\",\n \"This is due to the fact that there are 3 plates, that the holes have 2×3=6 sides, and that the directions of the movement are along the midperpendiculars of the hexagons.\",\n \"In a version also making it possible to modify the density of pinholes, a fourth plate is necessary.\",\n \"This plate is depicted in FIG.\",\n \"15.The set of modifiable pinholes obtained by means of three plates and depicted in FIG.\",\n \"13 in the reference position constitutes a first intermediate set.\",\n \"The plate in FIG.\",\n \"14 constitutes a second intermediate set.\",\n \"When the hole 1010 in the plate in FIG.\",\n \"15 is superimposed on the hole 1001 in the first intermediate set in FIG.\",\n \"13, the density of pinholes is at a maximum.\",\n \"When the hole 1013 in the plate in FIG.\",\n \"15 is superimposed on the hole 1001 in the first intermediate set in FIG.\",\n \"13, the number of pinholes per unit surface area is divided by 4.When the hole 1012 in the plate in FIG.\",\n \"15 is superimposed on the hole 1001 in the first intermediate set in FIG.\",\n \"13, the number of pinholes per unit surface area is divided by 9.When the hole 1011 in the plate in FIG.\",\n \"15 is superimposed on the hole 1001 in the first intermediate set in FIG.\",\n \"13, the number of pinholes per unit surface area is divided by 16.The three identical plates depicted in FIG.\",\n \"13 can be driven by means of an iris diaphragm drive system depicted in FIGS.\",\n \"16 and 17.The plates in FIG.\",\n \"13 are the plates 1040, 1041, 1042 depicted in FIG.\",\n \"17.These plates are metallic sheets tensioned over circular holding rings 1022, 1021, 1020 and carrying holes produced for example by laser piercing.\",\n \"These three fixing rings are connected to two control rings 1023, 1024.For example, the internal ring 1020 is connected to the control ring 1023 by a bar 1028 turning freely about an axis 1029 fixed in the control ring 1023.The internal ring 1020 is connected to the control ring 1024 by a bar 1026 turning freely about an axis 1027 fixed in the control ring 1024.The housing 1031 of the axis 1027 is oversized so as to be able to combine a rotation and translation with respect to the axis 1027.A tie rod 1030 is used for keeping the housing 1031 in abutment on the axis 1027.The other two holding rings are connected in a similar manner to the control rings.\",\n \"When the two control rings turn simultaneously in opposite directions and by an equal angle, the three plates move in translation with an angle of 120 degrees between the directions of each movement axis, as indicated in FIG.\",\n \"14.FIG.\",\n \"17 also depicts the fourth plate 1051 which is a sheet tensioned on a holding ring 1050.The holding ring 1050 is mounted on a two-axis positioner.\",\n \"The plates 1051 and 1042 are themselves pressed on thick plates of glass 1052 and 1043.When the whole of the system is in position the two glass plates 1052 and 1043 prevent deformations of the sheets (plates) 1042, 1041, 1040, 1051 carrying pinholes.\",\n \"Method of Guiding and Positioning the Plates In certain embodiments the plates carrying pinholes move in translation with respect to each other along a single axis.\",\n \"For example, in the first embodiment, the second and fourth embodiments, and also in the fifth embodiment with regard to the three plates moving by means of an iris diaphragm mechanism.\",\n \"This solution simplifies the system in that each plate moves with respect to another along a single axis.\",\n \"As indicated above a guide rail can be used for guiding the plates.\",\n \"However, a macroscopic guide rail is difficult to produce with the required precision.\",\n \"In order to obtain good positioning of the plates it is possible to replace such a guide rail with a set of microscopic guide rails.\",\n \"FIGS.\",\n \"18 to 20 illustrate the embodiment of a set of microscopic guide rails in a system with two plates of the type described in the first embodiment.\",\n \"FIG.\",\n \"18 shows the female part 1101, 1102, 1103 of the rails, produced by lithography in the same way as the pinholes, on the movable plate.\",\n \"FIG.\",\n \"19 shows the male part 1111, 1112, 1113 of the rails, produced by lithography on the fixed plate, but by means of a supplementary metallic layer.\",\n \"FIG.\",\n \"20 shows in section the assembly of the rails 1103 and 1113.The cross-section is produced along a cutting axis 1114 of the fixed plate and along a cutting axis 1104 of the movable plate, these two axes being deemed to be superimposed in the position depicted.\",\n \"The fixed glass plate 1120 carries a metallic layer 1122 leaving the pinhole 1115 free.\",\n \"It also carries a supplementary metallic layer 1113 constituting the male part of the guidance system.\",\n \"This male part is produced by lithography and the remains of a layer of resin 1125 have also been shown.\",\n \"The object of this layer of resin is to protect the bottom metallic layer 1122 when the rail 1113 is formed.\",\n \"The movable plate 1120 carries a metallic layer 1123 leaving free the pinhole 1105 and the female part 1103 of the guide rail.\",\n \"The space 1123 included between the two plates is filled with a lubricating liquid for preventing solid friction.\",\n \"The male part 1113 fits in the groove 1103 in order to guide the movement.\",\n \"The superimposition of the pinholes 1115 and 1105 produces a pinhole of reduced size.\",\n \"This guidance method can be adapted to the case where the plates are metallic sheets, as in the fifth embodiment.\",\n \"In this case male or female rails can be produced on each side of each sheet.\",\n \"The glass plates are then replaced by the metallic sheets.\",\n \"The diagram in FIG.\",\n \"21 is equivalent to that in FIG.\",\n \"20 but illustrates the case of plates formed by fine metallic sheets.\",\n \"The plate 1134 carries the male rail 1132 separated from the plate by a layer of protective resin 1133 used for producing the rail 1132 by lithography.\",\n \"The plate 1137 carries the female rail 1138 produced by lithography in a metallic layer 1135 separated from the plate by a resin 1139.The space between the two plates is filled with a lubricating liquid.\",\n \"A guide rail has been shown only on one side of each plate but it is possible to produce one on each side of each plate.\",\n \"In the case of the fifth embodiment, the guide rails which separate each metallic sheet guided by two adjacent sheets participate in the maintenance of the shape of the metallic sheets and in the prevention of deformations.\",\n \"Naturally sheet number 1051 in FIG.\",\n \"17 can be guided by this method only if its movement is restricted to a single direction.\",\n \"Industrial Applications The present set of pinholes can be used in a confocal microscope with single-point or multipoint illumination or in a confocal microscope intended to alternate the two illumination modes.\",\n \"For example, if a set of pinholes of the type described in the fourth embodiment replaces the set of pinholes used in the system described by FIG.\",\n \"1 of U.S. Pat.\",\n \"No.\",\n \"5,239,178 it becomes possible to modify the size and number of these pinholes and possibly to alternate between a multipoint and a single-point operating mode.\",\n \"Likewise, the array of pinholes in the fourth embodiment of the present invention can replace, with the same effect, the array of pinholes used in FIG.\",\n \"3 of the U.S. Pat.\",\n \"No.\",\n \"5,978,095.By using a modifiable array of pinholes according to the present invention in the microscope described by one of the first two embodiments of French patent application number 0103860 of Mar.\",\n \"22, 2001, it is possible to easily modify the diameter of the pinholes or their number, which affects the speed/resolution or speed/penetration depth compromise in the sample.\",\n \"The pinhole described in one of the first two embodiments of the present invention can also replace the interchangeable pinholes or the “iris” diaphragms normally used in single-point confocal microscopes with laser scanning.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10474269"}}},{"rowIdx":793,"cells":{"text":{"kind":"list like","value":[["Patent application cmos imager for cellular applications and methods of using such","Systems, methods and devices related to detecting and transmitting images.","Imaging systems and devices, as well as methods of using such that are provided herein include flicker detection and/or correction; and/or built-in self test associated with various analog circuitry in the imaging devices; and/or power reduction ability; and/or pixels with charge evacuation functionality; and/or a parallel to serial conversion unit and associated serial output interface; and/or other advanced functionality."],["1.A CMOS imaging device, the CMOS imaging device comprising: a pixel array; a selector, wherein the selector is operable to select between an input derived from the pixel array and at least one reference input, and wherein the selection between the input derived from the pixel array and the at least one reference input is at least in part based on a signal derived from a digital domain; and an analog to digital converter, wherein the analog to digital converter is operable to convert a signal derived from the selector to the digital domain.","2.The CMOS imaging device of claim 1, wherein the signal derived from the selector is an output of the selector that is passed through an analog processing circuit.","3.The CMOS imaging device of claim 2, wherein the analog processing circuit consists of one or more circuits selected from a group consisting of: a level shift circuit, a programmable offset circuit, and a programmable gain amplifier.","4.The CMOS imaging device of claim 2, the CMOS imaging device further comprising: an analog processing selector, wherein the analog processing selector is operable to bypass one or more portions of the analog processing circuit.","5.The CMOS imaging device of claim 1, the CMOS imaging device further comprising: a comparator receiving an output from the analog to digital converter, wherein the comparator is selected from a group consisting of: a programmable hardware comparator, a programmable software comparator, and a hardwired comparator.","6.The CMOS imaging device of claim 1, wherein the pixel array includes at least one pseudo-pixel, the CMOS imaging device further comprising: a pixel array selector, wherein the pixel array selector is operable to select the at least one pseudo-pixel, and wherein the input derived from the pixel array is derived from the pseudo-pixel.","7.The CMOS imaging device of claim 6, the CMOS imaging device further comprising: a column amplifier, wherein the column amplifier receives an output from the pseudo-pixel, and amplifies the output from the pseudo-pixel; and wherein the input derived from the pixel array is derived via the column amplifier.","8.The CMOS imaging device of claim 1, wherein the reference input to the selector is a pixel under reset, the CMOS imaging device further comprising: a circuit for selecting the pixel under reset; and wherein the selector is operable to provide a differential pair consisting of the input derived from the pixel array and the pixel under reset.","9.The CMOS imaging device of claim 1, wherein at least one of the reference inputs to the selector is tied to a pad of the CMOS imaging device.","10.The CMOS imaging device of claim 1, wherein at least one of the reference inputs to the selector comprises one or more voltages generated internal to the CMOS imaging device.","11.The CMOS imaging device of claim 1, wherein the input derived from the pixel array and the at least one reference input are both differential inputs.","12.A method for testing analog functionality of a CMOS imaging device in the digital domain, the method comprising: providing a pixel array; providing a selector, wherein the selector is operable to select between an input derived from the pixel array and at least one reference input; providing an analog to digital converter, wherein the analog to digital converter is operable to convert a signal derived from the selector to the digital domain; asserting a selection signal in the digital domain, wherein a derivative of the selection signal is input to the selector and at least in part controls the selection between the input derived from the pixel array and the reference input; and analyzing an output of the analog to digital converter to verify that the CMOS imaging device is functional.","13.The method of claim 12, wherein the reference input is a differential input, and wherein the reference input is a combination of two signals selected from a group consisting of: a common voltage, a negative reference voltage, and a positive reference voltage.","14.The method of claim 13, wherein asserting the selection signal causes the reference input to be selected, the method further comprising: providing an analog processing circuit, wherein the analog processing circuit receives the reference input from the selector, and processes the reference input, and wherein the signal derived from the selector is the processed reference input.","15.The method of claim 14, wherein the analog processing circuit comprises a first circuit and a second circuit, the method further comprising: bypassing the first circuit.","16.The method of claim 12, wherein the analyzing the output of the analog to digital converter comprises comparing the output of the analog to digital converter to determine if the output is within a predefined range.","17.The method of claim 12, wherein the pixel array includes at least one pseudo-pixel, the method further comprising: selecting the pseudo-pixel, wherein an output of the pseudo pixel is not sensitive to an illumination of the CMOS imaging device, and wherein the input derived from the pixel array is derived from the output of the pseudo-pixel; and wherein asserting the selection signal causes the input from the pixel array to be selected.","18.The method of claim 17, the method further comprising: providing a column amplifier, wherein the input derived from the pixel array is derived via the column amplifier.","19.The method of claim 12, wherein the reference input is a differential input, and wherein at least one differential signal of the reference input is generated external to the CMOS imaging device.","20.The method of claim 12, wherein the input derived from the pixel array is a differential input, wherein the at least one reference input comprises a reset level, and wherein the selection between the input derived from the pixel array and the reference input includes selecting one of the differential signals associated with the input derived from the pixel array and the reset level, and providing the input derived from the pixel array and the reset level as a differential pair.","21.The method of claim 20, the method further comprising: resetting at least one pixel in the pixel array; and selecting the at least one reset pixel, wherein the input derived from the pixel array is derived from the at least one reset pixel.","22.A CMOS imager, wherein the CMOS imager comprises: an image sensor, wherein the image sensor includes one or more rows of pixels, and wherein each of the pixels provides a pixel value; a storage element; a summing circuit, wherein the summing circuit is operable to sum the pixel values for each of the one or more rows to create row sums, and to store the row sums to the storage element; and a programmable core, wherein the programmable core includes instructions executable by the programmable core to: determine a first energy value and a second energy value associated with the row sums; compare the first energy value and the second energy value to estimate a flicker frequency; and constrain an exposure duration associated with the image sensor to approximate a multiple of the flicker duration.","23.The CMOS imager of claim 22, wherein the summing circuit is further operable to calculate row sums for a plurality of frames, wherein each of the plurality of frames includes pixel values from the two or more rows of pixels, and wherein the instructions are further executable by the programmable core to: determine a first energy value and a second energy value associated with the row sums for each frame; calculate a variance between first energy values across the plurality of frames; calculate a variance between second energy values across the plurality of frames; and wherein the comparing the first energy value and the second energy value to estimate a flicker frequency comprises comparing the variance of the first energy values with the variance of the second energy values.","24.The CMOS imager of claim 23, wherein the instructions are further executable by the programmable core to: compare the variance of the first energy values with the variance of the second energy values, wherein the larger is a greater variance; and comparing the greater variance with a predefined threshold, wherein the constraining the exposure frequency is done when the greater variance is greater than the predefined threshold.","25.The CMOS imager of claim 24, wherein the CMOS imager further comprises: a confidence counter, wherein the confidence counter is incremented whenever the estimated flicker frequency is approximately a first frequency, and decremented whenever the estimated flicker frequency is not approximately the first frequency.","26.The CMOS imager of claim 25, wherein the constraining the exposure frequency is done when the confidence counter changes its sign.","27.The CMOS imager of claim 22, wherein the CMOS imager further comprises: an interrupt associated with the summing circuit, wherein assertion of the interrupt indicates that summing for the one or more rows of pixels is complete.","28.The CMOS imager of claim 27, wherein the instructions are further executable by the programmable core to receive the interrupt.","29.The CMOS imager of claim 22, wherein the summing circuit is further operable to calculate row sums for a plurality of frames, wherein each of the plurality of frames includes pixel values from the two or more rows of pixels, and wherein the CMOS imager further comprises: an interrupt associated with the summing circuit, wherein assertion of the interrupt indicates that summing for the two or more rows of pixels for a frame is complete; and wherein the instructions are further executable by the programmable core to: receive the interrupt; determine a first energy value and a second energy value associated with the row sums for each frame; determine if a sufficient number of frames have been received to calculate a variance between first energy values and a variance between second energy values; calculate the variance between first energy values across the plurality of frames; calculate the variance between second energy values across the plurality of frames; and wherein the comparing the first energy value and the second energy value to estimate a flicker frequency comprises comparing the variance of the first energy values with the variance of the second energy values.","30.The CMOS imager of claim 22, wherein determining the first energy value and the second energy value comprises calculating an exact Fourier component for the first energy value and for the second energy value.","31.A method for automatically detecting flicker in an imaging device, the method comprising: providing an imaging device, wherein the imaging device includes an image sensor with two or more rows of pixels, and wherein each of the pixels provides a pixel value; retrieving at least a portion of the two or more rows of pixels, and summing the pixel values to create a row sum for each of the rows of pixels; determining a first energy value and a second energy value associated with the row sums; and comparing the first energy value and the second energy value to estimate a flicker frequency.","32.The method of claim 31, the method farther comprising: constraining an exposure duration associated with the image sensor to approximate a multiple of the flicker frequency period.","33.The method of claim 31, wherein determining the first energy value and the second energy value comprises calculating an approximate Fourier component for the first energy value and for the second energy value.","34.The method of claim 31, wherein the retrieving at least a portion of each of the two or more rows of pixels, sunning the pixel values, and determining a first energy value and a second energy value is performed for each of a plurality of frames, the method further comprising: calculating a variance between first energy values across the plurality of frames; calculating a variance between second energy values across the plurality of frames; and wherein the comparing the first energy value and the second energy value to estimate a flicker frequency comprises comparing the variance of the first energy values with the variance of the second energy values.","35.The method of claim 34, the method further comprising: comparing the variance of the first energy values with the variance of the second energy values, wherein the larger is a greater variance; and comparing the greater variance with a predefined threshold, wherein the constraining the exposure duration is done when the greater variance is greater than the predefined threshold.","36.The method of claim 35, the method further comprising: incrementing a confidence count whenever the estimated flicker frequency is approximately a first frequency, and decrementing the confidence count whenever the estimated flicker frequency is not approximately the first frequency.","37.The method of claim 36, wherein the constraining the exposure duration is done when the confidence counter changes sign.","38.The method of claim 31, wherein the summing the pixel values to create a row sum for each of the rows of pixels is done by a summing circuit included in the imaging device, the method further comprising: issuing an interrupt associated with the summing circuit, wherein the interrupt indicates that summing for the two or more rows of pixels is complete.","39.The method of claim 38, the method further comprising: receiving the interrupt, wherein determining the first and the second energy values is at least in part triggered by the interrupt.","40.An imaging device, wherein the imaging device comprises: a pixel array, wherein the pixel array comprises a plurality of rows, wherein each of the plurality of rows comprises a plurality of pixels, and wherein each of the plurality of pixels provides a pixel value; a summing circuit, wherein the summing circuit is operable to sum the pixel values for one or more of the plurality of rows to create row sums; an interrupt associated with the summing circuit, wherein the interrupt indicates that summing is complete for one or more of the plurality of rows; and a programmable core, wherein the programmable core includes instructions executable by the programmable core to: receive the interrupt; determine a first energy value and a second energy value associated with the row sums for each frame of a plurality of frames; calculate a variance between first energy values across the plurality of frames; calculate a variance between second energy values across the plurality of frames; and compare the variance of the first energy values with the variance of the second energy values, wherein the larger is a greater variance.","41.The imaging device of claim 40, wherein the instructions are further executable by the programmable core to: constrain the exposure duration to a multiple of the flicker frequency period.","42.A CMOS imager, wherein the CMOS imager comprises: an image sensor; a sub-sample control system, wherein the sub-sample control system is operable to identify one or more signals from the image sensor that are not processed; an analog processing circuit, wherein the analog processing circuit receives the one or more signals from the image sensor at one or more inputs; and a processing control system, wherein the processing control system is operable to limit switching of the one or more inputs.","43.The CMOS imager of claim 42, wherein the analog processing circuit comprises a column amplifier, and wherein the one or more inputs are inputs to the column amplifier.","44.The CMOS imager of claim 42, wherein the analog processing circuit comprises a level shift circuit, and wherein the one or more inputs are inputs to the level shift circuit.","45.The CMOS imager of claim 42, wherein the analog processing circuit comprises a programmable offset circuit, and wherein the one or more inputs are inputs to the programmable offset circuit.","46.The CMOS imager of claim 42, wherein the processing control system is operable to maintain a common mode feedback loop of an amplifier associated with the analog processing circuit in an active state, while limiting switching of the one or more inputs.","47.The CMOS imager of claim 42, the CMOS imager further comprising: a digital processing circuit, wherein the processing control system is further operable to disable the digital processing circuit.","48.A method for processing images in a power sensitive application, the method comprising: providing a CMOS imager, wherein the CMOS imager includes a pixel array, and an analog processing circuit; defining a first portion and a second portion of the pixel array, wherein the first portion of the pixel array includes a output image area and the second portion of the pixel array includes a dropped image area; receiving a pixel signal from the second portion of the pixel array; and placing the analog processing circuit in a standby mode, wherein the pixel signal from the second portion of the pixel array is not processed.","49.The method of claim 48, wherein placing the analog processing circuit in the standby mode comprises: limiting switching of one or more inputs to the analog processing circuit.","50.The method of claim 48, wherein placing the analog processing circuit in the standby mode comprises: electrically coupling one or more inputs of the analog processing circuit to a static voltage, wherein switching of the analog processing circuit is reduced.","51.The method of claim 48, wherein placing the analog processing circuit in the standby mode comprises: electrically coupling one or more inputs of the analog processing circuit to a static voltage, while maintaining switching associated with a common mode feedback loop of one or more amplifiers associated with the analog processing circuit, wherein switching of the analog processing circuit is reduced, and bias currents are reduced while maintaining the one or more amplifiers at its inertial state.","52.The method of claim 48, the method further comprising: receiving a pixel signal from the first portion of the pixel array; and placing the analog processing circuit in an operational mode, wherein the pixel signal from the first portion of the pixel array is processed.","53.The method of claim 48, wherein the power sensitive application comprises video transmission via a cellular telephone.","54.The method of claim 48, wherein the power sensitive application comprises image detection and processing using a mobile device.","55.The method of claim 54, wherein the mobile device is selected from a group consisting of a cellular telephone, a personal digital assistant, and a laptop computer.","56.The method of claim 48, wherein the CMOS imager further comprises a digital processing circuit, the method further comprising: disabling the digital processing circuit, wherein the pixel signal from the second portion of the pixel array is not processed.","57.The method of claim 56, wherein disabling the digital processing circuit comprises: gating at least one clock driving the digital processing circuit.","58.The method of claim 52, wherein the CMOS imager further comprises a digital processing circuit, the method further comprising: enabling the digital processing circuit, wherein the pixel signal from the first portion of the pixel array is processed.","59.The method of claim 48, wherein the analog processing circuit comprises: a column amplifier; and a level shift circuit.","60.The method of claim 59, wherein the analog processing circuit further comprises: a black level offset; a programmable gain amplifier; and an ADC.","61.The method of claim 56, wherein the digital processing circuit includes a translation unit.","62.A method for reducing power consumption in an imaging device, the method comprising: providing an imaging device including an image sensor and an analog processing circuit; sub-sampling the image sensor, wherein a first portion of the image sensor is not processed; receiving an image signal from the first portion of the image sensor; and placing the analog processing circuit in a standby state, wherein the image signal from the first portion is not processed.","63.The method of claim 62, wherein placing the analog processing circuit in a standby state comprises: limiting switching of one or more inputs to the analog processing circuit.","64.The method of claim 62, wherein the analog processing circuit comprises: a level shift circuit; a programmable offset circuit; and a programmable gain amplifier.","65.The method of claim 64, wherein placing the analog processing circuit in a standby state comprises: electrically coupling one or more inputs of the analog processing circuit to a static voltage, wherein switching of the analog processing circuit is reduced.","66.The method of claim 65, wherein placing the analog processing circuit in a standby state further comprises: maintaining switching associated with a common mode feedback loop of one or more amplifiers associated with the analog processing circuit, wherein the one or more amplifiers are maintained in an inertial state, and a bias current associated with the amplifiers is reduced.","67.The method of claim 62, wherein sub-sampling the image sensor is done to create a bandwidth matched image output from the imaging device.","68.The method of claim 62, the method further comprising: receiving an image signal from a second portion of the image sensor, wherein the second portion the image sensor is processed; and placing the analog processing circuit in an operational mode, wherein the image signal from the second portion is processed.","69.The method of claim 68, wherein placing the analog processing circuit in an operational mode comprises electrically coupling one or more inputs of the analog processing circuit to the image signal from the second portion.","70.The method of claim 62, wherein the imaging device further comprises a digital processing circuit, the method further comprising: disabling the digital processing circuit.","71.A CMOS imager, wherein the CMOS imager comprises an image sensor; a parallel to serial data conversion unit, wherein the parallel to serial data conversion unit receives an image derived from the image sensor and converts the image to a serial data stream; and a serial output interface, wherein the serial output interface provides access to the serial data stream.","72.The CMOS imager of claim 71, wherein the image sensor comprises one or more pixels, and wherein the one or more pixels include a charge evacuation element.","73.The CMOS imager of claim 71, wherein the serial output interface includes a clock signal and a data signal, and wherein the clock signal is only active when valid data is presented on the data signal.","74.The CMOS imager of claim 71, wherein the serial output includes a clock signal, a data signal, and a qualifying signal, and wherein the qualifying signal indicates the presence of valid data on the data signal."],["<SOH> BACKGROUND OF THE INVENTION <EOH>The present invention relates generally to systems and methods for detecting and/or transmitting images.","More particularly, the present invention relates to detecting and transmitting images in relation to a cellular telephone.","Image processors typically detect light reflected off of a subject, and convert the detected light to an electrical image.","The electrical image is then sampled at a frequency that is the reciprocal of the integration time.","In many cases, such a frequency is different from the frequency of light reflected off the subject.","In such cases, a modulation of the displayed image can result.","Such modulation appears as a flicker and can be very distracting.","To overcome this problem, some image processors allow for manual selection between different integration times that allow a user to reduce the flicker.","For example, where an image processor will be used in a 60 Hz and a 50 Hz lighting environment, a manual selection between an integration time associated with either 60 Hz or 50 Hz can be provided.","This alleviates some flicker, but requires adjustment from a user.","Furthermore, such an approach is limited to reducing flicker in a limited number of pre-determined environments, and thus may not be able to address various flicker situations.","This is particularly problematic for mobile devices that are used in ever changing environments.","Further, a limited reservoir of power typically exists for mobile devices.","As such, power consuming imaging applications are often incompatible with mobile devices.","Various approaches exist to increase the amount of power available to mobile devices, such that the power requirements of an imaging application are not prohibitive.","However, such approaches often are both expensive and can result in increased dimensions of a mobile device.","In some cases, light detected by an image processor is detected by a pixel array and a variety of analog processing circuitry.","Output from the pixel array representing the amount of detected light is processed by analog circuitry to produce an image which is subsequently converted from the analog domain to the digital domain.","The digital image is subsequently provided to a monitor of some sort for viewing.","Testing such an imaging device can include reflecting light off a known image, and determining if the image was properly acquired and/or processed by the imaging device.","This method provides an effective approach for testing such devices, however, the method is subject to a number of variables including lighting and image stability which can effect testing of the imaging device.","Providing such a test, and controlling for such variables can be costly and time consuming.","Further, such an approach tests the imaging device holistically, and is limited in its ability to identify sub-components of the imaging device which may have failed.","Hence, for at least the aforementioned reasons, there exists a need in the art to provide advanced systems, methods and devices for detecting and/or transmitting images."],["<SOH> BRIEF SUMMARY OF THE INVENTION <EOH>The present invention provides systems, devices, and methods for imaging.","More particularly, the present invention provides devices and methods for capturing images, formatting the images, and/or transmitting the images via a cellular telephone network or other wireless network.","Various aspects of the present invention include an imaging device with one or more of the following units: a control unit, a serial interface unit, a parallel interface unit, an imaging unit, and a translation unit.","In some embodiments, flicker detection and/or correction functionality is incorporated.","Additionally, in some embodiments, a built-in self test associated with various analog circuitry in the imaging device is included.","Further, in some embodiments, systems and methods for reducing power consumption by various analog circuitry within the imaging device are included.","Yet further, in some embodiments, a parallel to serial conversion unit and associated serial output interface are included.","These and many other novel aspects of the present invention are provided in greater detail in the proceeding portions of this document.","One particular embodiment of the present invention provides an imaging device and methods for using such that provide for advanced testing ability.","In one aspect, an imaging device includes a pixel array.","The imaging device further includes a selector that is operable to select between an input derived from the pixel array and at least one reference input.","Such selection between the input derived from the pixel array and the at least one reference input is based at least in part on a signal derived from a digital domain.","Further, the imaging device includes an analog to digital converter that is operable to convert a signal derived from the selector to the digital domain.","In some cases, such an embodiment provides an ability to verify the functionality of analog circuitry within the imaging device using a digital tester.","In some aspects of the embodiment, the output of the selector is passed through an analog processing circuit that can include one or more circuits selected from a group consisting of: a level shift circuit, a programmable offset circuit, and a programmable gain amplifier.","In various aspects of the embodiment, the imaging device further comprises an analog processing selector that is operable to bypass one or more portions of the analog processing circuit.","In other aspects, the imaging device further includes a comparator receiving an output from the analog to digital converter.","Such a comparator is selected from a group consisting of: a programmable hardware comparator, a programmable software comparator, and a hardwired comparator.","In some aspects of the embodiment, the pixel array includes at least one pseudo-pixel, and the imaging device additionally includes a pixel array selector that is operable to select the pseudo-pixel such that input derived from the pixel array is derived from the pseudo-pixel.","Other aspects include a column amplifier that receives and amplifies the output from the pseudo-pixel.","In some aspects, the reference input to the selector is a reset level that includes a pixel under reset condition, and the imaging device farther includes a circuit for selecting the reset level and pairing it with an output from the pixel array.","In this way, the selector is operable to provide a differential pair consisting of the input derived from the pixel array and the reset level.","In other aspects, the reference input to the selector is tied to a pad of the imaging device, and in yet other aspects, the reference input to the selector comprises one or more voltages generated internal to the imaging device.","In other embodiments of the present invention, imaging devices and methods for using such are provided to detect and/or reduce image flicker.","Such an imaging device includes an image sensor with one or more rows of pixels, where each of the pixels provides a pixel value.","The device further includes a storage element and a summing circuit.","Such a summing circuit can be a hardware summing circuit, either programmable or hardwired, or a software summing circuit including for example a programmable core.","The summing circuit is operable to sum the pixel values for each of the one or more rows to create row sums, and to store the row sums to the storage element.","In addition, the imaging device includes a programmable core that includes instructions executable by the programmable core to: determine a first energy value and a second energy value associated with the row sums; compare the first energy value and the second energy value to estimate a flicker frequency; and constrain an exposure duration associated with the image sensor to approximate a multiple of the flicker frequency.","Such a programmable core can also be used for the summing circuit.","In some aspects of the embodiment, the summing circuit is further operable to calculate row sums for a plurality of frames, where each of the plurality of frames includes pixel values from one or more rows of pixels, and where the instructions are further executable by the programmable core to: determine a first energy value and a second energy value associated with the row sums for each frame; calculate a variance between first energy values across the plurality of frames; calculate a variance between second energy values across the plurality of frames; and wherein the comparing the first energy value and the second energy value to estimate a flicker frequency comprises comparing the variance of the first energy values with the variance of the second energy values.","In yet other aspects of the embodiment, the instructions are further executable by the programmable core to: compare the variance of the first energy values with the variance of the second energy values, and to compare the greater variance with a predefined threshold where constraining the exposure frequency is done when the greater variance is greater than the predefined threshold.","In various aspects, a confidence counter is implemented to control switching from an exposure frequency to a multiple of an estimated flicker frequency.","In such aspects, a change of frequencies can be limited to situations where the confidence counter changes sign.","In one particular aspect of the embodiment, the summing circuit is further operable to calculate row sums for a plurality of frames, wherein each of the plurality of frames includes pixel values from the two or more rows of pixels.","Further, the imaging device further includes an interrupt associated with the summing circuit, wherein assertion of the interrupt indicates that summing for the two or more rows of pixels for a frame is complete; and wherein the instructions are further executable by the programmable core to: receive the interrupt; determine a first energy value and a second energy value associated with the row sums for each frame; determine if a sufficient number of frames have been received to calculate a variance between first energy values and a variance between second energy values; calculate the variance between first energy values across the plurality of frames; calculate the variance between second energy values across the plurality of frames; and wherein the comparing the first energy value and the second energy value to estimate a flicker frequency comprises comparing the variance of the first energy values with the variance of the second energy values.","Some aspects of the invention provide a method for automatically detecting flicker in an imaging device.","The method includes providing an imaging device, wherein the imaging device includes an image sensor with two or more rows of pixels, and wherein each of the pixels provides a pixel value.","In addition, the method includes retrieving at least a portion of the two or more rows of pixels, and summing the pixel values to create a row sum for each of the rows of pixels; and determining a first energy value and a second energy value associated with the row sums.","The first and second energy values are compared to estimate a flicker frequency.","Various embodiments of the present invention include a pixel with a charge evacuation mechanism for reducing noise evident at the output of the pixel.","In one aspect, the pixel includes a reset element used in relation to charging a light sensitive element.","The evacuation mechanism is used to dissipate charge build up about the reset element.","Dissipation of the charge results in a reduction of noise at the output of the pixel.","In particular embodiments, such a pixel and/or charge evacuation mechanism is implemented using complementary metal oxide semiconductor (“CMOS”), field effect transistor (“FET”) technology.","Further, aspects of the present invention include systems and methods for utilizing such pixels.","More particularly, such methods include approaches for applying various control signals to the pixel and systems include elements for producing the various control signals.","One particular aspect of the invention provides a MOS pixel that includes a reset transistor with a reset drain and a reset gate.","Further, the pixel includes a charge evacuation element that is electrically coupled to the reset drain and is operable to evacuate charge accumulated in a channel of the reset transistor.","In some embodiments, the charge evacuation element is a MOS transistor with a charge evacuation drain, a charge evacuation source, and a charge evacuation gate.","In particular cases, the charge evacuation source and the charge evacuation drain are both electrically coupled to the reset drain.","In various embodiments, the pixel further includes a light sensitive element electrically coupled to the reset drain.","Further embodiments comprise a source follower transistor that includes a source follower gate and a source follower drain, the source follower gate being electrically coupled to the reset drain.","Such embodiments further include a selection transistor with a selection drain, a selection source, and a selection gate, where the selection source is electrically coupled to the source follower drain.","A selection signal is electrically coupled to the source gate such that assertion thereof causes a representation of a signal from the node of the light sensitive element to be present on an output of the pixel.","In some embodiments, the charge evacuation element is a MOS transistor with a charge evacuation source and a charge evacuation drain, both of which are coupled to the reset drain.","The charge evacuation element further includes a charge evacuation gate.","Further, such embodiments can include a reset signal driving the reset gate and a complement of the reset signal driving the charge evacuation gate.","In some instances, the complement of the reset signal is delayed from the reset signal.","Other embodiments of the present invention provide a circuit including a reset transistor comprising a reset gate and a reset drain.","The circuit further includes a photodiode with a node of the photodiode electrically coupled to the reset drain, and a charge evacuation transistor that includes a charge evacuation gate, a charge evacuation drain, and a charge evacuation source.","The charge evacuation source and the charge evacuation drain are both electrically coupled to the reset drain and the charge evacuation transistor is operable to evacuate charge accumulated in a channel of the reset transistor.","Various embodiments of the circuit also include a source follower transistor comprised of a source follower gate and a source follower drain.","The source follower gate is electrically coupled to the reset drain.","the circuit further includes a selection transistor comprised of a selection source, a selection gate, and a selection drain.","The selection source is electrically coupled to the source follower drain.","A selection signal is provided that when asserted causes a representation of a signal from the photodiode to be present on an output of the pixel.","Yet other embodiments of the present invention provide an imaging system.","Such an imaging system includes a group of pixel elements.","One or more of the pixel elements includes a reset element, a charge evacuation element, and a light sensitive element.","In some cases, the imaging system further includes an optical device, wherein the optical device transfers light to the group of pixel elements, and wherein the light strikes the light sensitive element of the pixel elements.","In addition, a timing circuit is included that provides at least one control signal to the group of pixel elements.","In one particular instance, the group of pixel elements are arranged in a rectangular array.","Yet another embodiment of the present invention provides an image sensor including a plurality of pixel devices.","The plurality of pixel devices are arranged as a plurality of rows and a plurality of columns.","Each of the pixel devices includes: a light detecting element, and a charge evacuation element for dissipating unwanted charge built up in the image sensor.","Yet additional embodiments provide methods for detecting an image.","Such methods include providing a pixel device that comprises a charge evacuation element, and applying a charge evacuation control signal to the charge evacuation element, wherein a charge accumulation in a channel of a reset transistor of the pixel device is reduced.","Other embodiments provide systems and method for reducing power consumption in an imaging device.","In one aspect, an imaging device is included with an image sensor and a sub-sample control system.","The sub-sample control system is operable to identify one or more signals from the image sensor that are not processed.","The imaging device further includes an analog processing circuit that receives the one or more signals from the image sensor at one or more inputs, and a processing control system that is operable to limit switching of the one or more inputs.","In some aspects, the analog processing circuit comprises a column amplifier that receives the one or more inputs.","In various aspects, the analog processing circuit comprises a level shift circuit, and/or a programmable or black offset circuit.","In some aspects, the processing control system is operable to maintain a common mode feedback loop of an amplifier associated with the analog processing circuit in an active state, while limiting switching of the one or more inputs.","In other aspects, the imaging device further includes a digital processing circuit, and the processing control system is further operable to disable the digital processing circuit.","Yet other embodiments of the present invention provide a CMOS imager that includes an image sensor, and a parallel to serial data conversion unit.","The parallel to serial data conversion unit receives an image derived from the image sensor and converts the image to a serial data stream.","The serial data stream is output via a serial output interface.","In some aspects of the embodiment, the image sensor comprises one or more pixels, and wherein the one or more pixels include a charge evacuation element.","in other aspects, the serial output interface includes a clock signal and a data signal.","The clock signal is only active when valid data is presented on the data signal.","In yet other aspects, the serial output includes a clock signal, a data signal, and a qualifying signal.","The qualifying signal indicates the presence of valid data on the data signal.","These and other aspects are more fully developed in the detailed description below.","Thus, the summary provides only a general outline of the embodiments according to the present invention.","Many other objects, features and advantages of the present invention will become more fully apparent from the following detailed description, the appended claims and the accompanying drawings."],["CROSS-REFERENCES TO RELATED APPLICATIONS This application claims the benefit of U.S.","Provisional Patent Application No.","60/294,388, filed on May 29, 2001, which is incorporated herein by reference in its entirety for all purposes.","BACKGROUND OF THE INVENTION The present invention relates generally to systems and methods for detecting and/or transmitting images.","More particularly, the present invention relates to detecting and transmitting images in relation to a cellular telephone.","Image processors typically detect light reflected off of a subject, and convert the detected light to an electrical image.","The electrical image is then sampled at a frequency that is the reciprocal of the integration time.","In many cases, such a frequency is different from the frequency of light reflected off the subject.","In such cases, a modulation of the displayed image can result.","Such modulation appears as a flicker and can be very distracting.","To overcome this problem, some image processors allow for manual selection between different integration times that allow a user to reduce the flicker.","For example, where an image processor will be used in a 60 Hz and a 50 Hz lighting environment, a manual selection between an integration time associated with either 60 Hz or 50 Hz can be provided.","This alleviates some flicker, but requires adjustment from a user.","Furthermore, such an approach is limited to reducing flicker in a limited number of pre-determined environments, and thus may not be able to address various flicker situations.","This is particularly problematic for mobile devices that are used in ever changing environments.","Further, a limited reservoir of power typically exists for mobile devices.","As such, power consuming imaging applications are often incompatible with mobile devices.","Various approaches exist to increase the amount of power available to mobile devices, such that the power requirements of an imaging application are not prohibitive.","However, such approaches often are both expensive and can result in increased dimensions of a mobile device.","In some cases, light detected by an image processor is detected by a pixel array and a variety of analog processing circuitry.","Output from the pixel array representing the amount of detected light is processed by analog circuitry to produce an image which is subsequently converted from the analog domain to the digital domain.","The digital image is subsequently provided to a monitor of some sort for viewing.","Testing such an imaging device can include reflecting light off a known image, and determining if the image was properly acquired and/or processed by the imaging device.","This method provides an effective approach for testing such devices, however, the method is subject to a number of variables including lighting and image stability which can effect testing of the imaging device.","Providing such a test, and controlling for such variables can be costly and time consuming.","Further, such an approach tests the imaging device holistically, and is limited in its ability to identify sub-components of the imaging device which may have failed.","Hence, for at least the aforementioned reasons, there exists a need in the art to provide advanced systems, methods and devices for detecting and/or transmitting images.","BRIEF SUMMARY OF THE INVENTION The present invention provides systems, devices, and methods for imaging.","More particularly, the present invention provides devices and methods for capturing images, formatting the images, and/or transmitting the images via a cellular telephone network or other wireless network.","Various aspects of the present invention include an imaging device with one or more of the following units: a control unit, a serial interface unit, a parallel interface unit, an imaging unit, and a translation unit.","In some embodiments, flicker detection and/or correction functionality is incorporated.","Additionally, in some embodiments, a built-in self test associated with various analog circuitry in the imaging device is included.","Further, in some embodiments, systems and methods for reducing power consumption by various analog circuitry within the imaging device are included.","Yet further, in some embodiments, a parallel to serial conversion unit and associated serial output interface are included.","These and many other novel aspects of the present invention are provided in greater detail in the proceeding portions of this document.","One particular embodiment of the present invention provides an imaging device and methods for using such that provide for advanced testing ability.","In one aspect, an imaging device includes a pixel array.","The imaging device further includes a selector that is operable to select between an input derived from the pixel array and at least one reference input.","Such selection between the input derived from the pixel array and the at least one reference input is based at least in part on a signal derived from a digital domain.","Further, the imaging device includes an analog to digital converter that is operable to convert a signal derived from the selector to the digital domain.","In some cases, such an embodiment provides an ability to verify the functionality of analog circuitry within the imaging device using a digital tester.","In some aspects of the embodiment, the output of the selector is passed through an analog processing circuit that can include one or more circuits selected from a group consisting of: a level shift circuit, a programmable offset circuit, and a programmable gain amplifier.","In various aspects of the embodiment, the imaging device further comprises an analog processing selector that is operable to bypass one or more portions of the analog processing circuit.","In other aspects, the imaging device further includes a comparator receiving an output from the analog to digital converter.","Such a comparator is selected from a group consisting of: a programmable hardware comparator, a programmable software comparator, and a hardwired comparator.","In some aspects of the embodiment, the pixel array includes at least one pseudo-pixel, and the imaging device additionally includes a pixel array selector that is operable to select the pseudo-pixel such that input derived from the pixel array is derived from the pseudo-pixel.","Other aspects include a column amplifier that receives and amplifies the output from the pseudo-pixel.","In some aspects, the reference input to the selector is a reset level that includes a pixel under reset condition, and the imaging device farther includes a circuit for selecting the reset level and pairing it with an output from the pixel array.","In this way, the selector is operable to provide a differential pair consisting of the input derived from the pixel array and the reset level.","In other aspects, the reference input to the selector is tied to a pad of the imaging device, and in yet other aspects, the reference input to the selector comprises one or more voltages generated internal to the imaging device.","In other embodiments of the present invention, imaging devices and methods for using such are provided to detect and/or reduce image flicker.","Such an imaging device includes an image sensor with one or more rows of pixels, where each of the pixels provides a pixel value.","The device further includes a storage element and a summing circuit.","Such a summing circuit can be a hardware summing circuit, either programmable or hardwired, or a software summing circuit including for example a programmable core.","The summing circuit is operable to sum the pixel values for each of the one or more rows to create row sums, and to store the row sums to the storage element.","In addition, the imaging device includes a programmable core that includes instructions executable by the programmable core to: determine a first energy value and a second energy value associated with the row sums; compare the first energy value and the second energy value to estimate a flicker frequency; and constrain an exposure duration associated with the image sensor to approximate a multiple of the flicker frequency.","Such a programmable core can also be used for the summing circuit.","In some aspects of the embodiment, the summing circuit is further operable to calculate row sums for a plurality of frames, where each of the plurality of frames includes pixel values from one or more rows of pixels, and where the instructions are further executable by the programmable core to: determine a first energy value and a second energy value associated with the row sums for each frame; calculate a variance between first energy values across the plurality of frames; calculate a variance between second energy values across the plurality of frames; and wherein the comparing the first energy value and the second energy value to estimate a flicker frequency comprises comparing the variance of the first energy values with the variance of the second energy values.","In yet other aspects of the embodiment, the instructions are further executable by the programmable core to: compare the variance of the first energy values with the variance of the second energy values, and to compare the greater variance with a predefined threshold where constraining the exposure frequency is done when the greater variance is greater than the predefined threshold.","In various aspects, a confidence counter is implemented to control switching from an exposure frequency to a multiple of an estimated flicker frequency.","In such aspects, a change of frequencies can be limited to situations where the confidence counter changes sign.","In one particular aspect of the embodiment, the summing circuit is further operable to calculate row sums for a plurality of frames, wherein each of the plurality of frames includes pixel values from the two or more rows of pixels.","Further, the imaging device further includes an interrupt associated with the summing circuit, wherein assertion of the interrupt indicates that summing for the two or more rows of pixels for a frame is complete; and wherein the instructions are further executable by the programmable core to: receive the interrupt; determine a first energy value and a second energy value associated with the row sums for each frame; determine if a sufficient number of frames have been received to calculate a variance between first energy values and a variance between second energy values; calculate the variance between first energy values across the plurality of frames; calculate the variance between second energy values across the plurality of frames; and wherein the comparing the first energy value and the second energy value to estimate a flicker frequency comprises comparing the variance of the first energy values with the variance of the second energy values.","Some aspects of the invention provide a method for automatically detecting flicker in an imaging device.","The method includes providing an imaging device, wherein the imaging device includes an image sensor with two or more rows of pixels, and wherein each of the pixels provides a pixel value.","In addition, the method includes retrieving at least a portion of the two or more rows of pixels, and summing the pixel values to create a row sum for each of the rows of pixels; and determining a first energy value and a second energy value associated with the row sums.","The first and second energy values are compared to estimate a flicker frequency.","Various embodiments of the present invention include a pixel with a charge evacuation mechanism for reducing noise evident at the output of the pixel.","In one aspect, the pixel includes a reset element used in relation to charging a light sensitive element.","The evacuation mechanism is used to dissipate charge build up about the reset element.","Dissipation of the charge results in a reduction of noise at the output of the pixel.","In particular embodiments, such a pixel and/or charge evacuation mechanism is implemented using complementary metal oxide semiconductor (“CMOS”), field effect transistor (“FET”) technology.","Further, aspects of the present invention include systems and methods for utilizing such pixels.","More particularly, such methods include approaches for applying various control signals to the pixel and systems include elements for producing the various control signals.","One particular aspect of the invention provides a MOS pixel that includes a reset transistor with a reset drain and a reset gate.","Further, the pixel includes a charge evacuation element that is electrically coupled to the reset drain and is operable to evacuate charge accumulated in a channel of the reset transistor.","In some embodiments, the charge evacuation element is a MOS transistor with a charge evacuation drain, a charge evacuation source, and a charge evacuation gate.","In particular cases, the charge evacuation source and the charge evacuation drain are both electrically coupled to the reset drain.","In various embodiments, the pixel further includes a light sensitive element electrically coupled to the reset drain.","Further embodiments comprise a source follower transistor that includes a source follower gate and a source follower drain, the source follower gate being electrically coupled to the reset drain.","Such embodiments further include a selection transistor with a selection drain, a selection source, and a selection gate, where the selection source is electrically coupled to the source follower drain.","A selection signal is electrically coupled to the source gate such that assertion thereof causes a representation of a signal from the node of the light sensitive element to be present on an output of the pixel.","In some embodiments, the charge evacuation element is a MOS transistor with a charge evacuation source and a charge evacuation drain, both of which are coupled to the reset drain.","The charge evacuation element further includes a charge evacuation gate.","Further, such embodiments can include a reset signal driving the reset gate and a complement of the reset signal driving the charge evacuation gate.","In some instances, the complement of the reset signal is delayed from the reset signal.","Other embodiments of the present invention provide a circuit including a reset transistor comprising a reset gate and a reset drain.","The circuit further includes a photodiode with a node of the photodiode electrically coupled to the reset drain, and a charge evacuation transistor that includes a charge evacuation gate, a charge evacuation drain, and a charge evacuation source.","The charge evacuation source and the charge evacuation drain are both electrically coupled to the reset drain and the charge evacuation transistor is operable to evacuate charge accumulated in a channel of the reset transistor.","Various embodiments of the circuit also include a source follower transistor comprised of a source follower gate and a source follower drain.","The source follower gate is electrically coupled to the reset drain.","the circuit further includes a selection transistor comprised of a selection source, a selection gate, and a selection drain.","The selection source is electrically coupled to the source follower drain.","A selection signal is provided that when asserted causes a representation of a signal from the photodiode to be present on an output of the pixel.","Yet other embodiments of the present invention provide an imaging system.","Such an imaging system includes a group of pixel elements.","One or more of the pixel elements includes a reset element, a charge evacuation element, and a light sensitive element.","In some cases, the imaging system further includes an optical device, wherein the optical device transfers light to the group of pixel elements, and wherein the light strikes the light sensitive element of the pixel elements.","In addition, a timing circuit is included that provides at least one control signal to the group of pixel elements.","In one particular instance, the group of pixel elements are arranged in a rectangular array.","Yet another embodiment of the present invention provides an image sensor including a plurality of pixel devices.","The plurality of pixel devices are arranged as a plurality of rows and a plurality of columns.","Each of the pixel devices includes: a light detecting element, and a charge evacuation element for dissipating unwanted charge built up in the image sensor.","Yet additional embodiments provide methods for detecting an image.","Such methods include providing a pixel device that comprises a charge evacuation element, and applying a charge evacuation control signal to the charge evacuation element, wherein a charge accumulation in a channel of a reset transistor of the pixel device is reduced.","Other embodiments provide systems and method for reducing power consumption in an imaging device.","In one aspect, an imaging device is included with an image sensor and a sub-sample control system.","The sub-sample control system is operable to identify one or more signals from the image sensor that are not processed.","The imaging device further includes an analog processing circuit that receives the one or more signals from the image sensor at one or more inputs, and a processing control system that is operable to limit switching of the one or more inputs.","In some aspects, the analog processing circuit comprises a column amplifier that receives the one or more inputs.","In various aspects, the analog processing circuit comprises a level shift circuit, and/or a programmable or black offset circuit.","In some aspects, the processing control system is operable to maintain a common mode feedback loop of an amplifier associated with the analog processing circuit in an active state, while limiting switching of the one or more inputs.","In other aspects, the imaging device further includes a digital processing circuit, and the processing control system is further operable to disable the digital processing circuit.","Yet other embodiments of the present invention provide a CMOS imager that includes an image sensor, and a parallel to serial data conversion unit.","The parallel to serial data conversion unit receives an image derived from the image sensor and converts the image to a serial data stream.","The serial data stream is output via a serial output interface.","In some aspects of the embodiment, the image sensor comprises one or more pixels, and wherein the one or more pixels include a charge evacuation element.","in other aspects, the serial output interface includes a clock signal and a data signal.","The clock signal is only active when valid data is presented on the data signal.","In yet other aspects, the serial output includes a clock signal, a data signal, and a qualifying signal.","The qualifying signal indicates the presence of valid data on the data signal.","These and other aspects are more fully developed in the detailed description below.","Thus, the summary provides only a general outline of the embodiments according to the present invention.","Many other objects, features and advantages of the present invention will become more fully apparent from the following detailed description, the appended claims and the accompanying drawings.","BRIEF DESCRIPTION OF THE DRAWINGS A further understanding of the nature and advantages of the present invention may be realized by reference to the figures which are described in remaining portions of the specification.","In the figures, like reference numerals are used throughout several figures to refer to similar components.","In some instances, a sub-label consisting of a lower case letter is associated with a reference numeral to denote one of multiple similar components.","When reference is made to a reference numeral without specification to an existing sub-label, it is intended to refer to all such multiple similar components.","FIG.","1 is a block diagram of a CMOS imager in accordance with various embodiments of the present invention; FIG.","2 is a timing diagram of frame rate control implemented in the CMOS imager of FIG.","1; FIG.","3 is a timing diagram illustrating the various clocks used in relation to the CMOS imager of FIG.","1; FIG.","4 is a block diagram of a sensor in accordance with one embodiment of the present invention; FIG.","5 is a block diagram of a virtual image window in accordance with various embodiments of the present invention; FIG.","6 illustrates a Bayer grid array; FIG.","7 illustrates a pixel circuit in accordance with various embodiments of the present invention; FIG.","8 is a flow diagram illustrating a method for using the pixel circuit of FIG.","7 in accordance with an embodiment of the present invention; FIG.","9 are timing diagrams illustrating the operation of the pixel of FIG.","7.FIG.","10 is an exemplary column amplifier circuit; FIGS.","11-13 are timing diagrams illustrating the operation of the column amplifier circuit of FIG.","10; FIG.","14 is a timing diagram illustrating the exposure and sensing of a flame of video in accordance with an embodiment of the present invention; FIG.","15 is a timing diagram illustrating sampling of a pixel array incorporated in the sensor of FIG.","4; FIG.","16 illustrates high gain and high dynamic range modes of the level shift circuit of FIG.","4; FIG.","17 illustrates the use of black level and gain controls in relation to the sensor of FIG.","4; FIG.","18 is a timing diagram of various control signals associated with the sensor of FIG.","4; FIG.","19 is a timing diagram illustrating the interaction of various standby mode control signals; FIG.","20 is a block diagram of a standby mode circuit in accordance with embodiments of the present invention; FIG.","21 illustrates an implementation of a standby mode in a switched capacitor circuit in accordance with embodiments of the present invention; FIG.","22 is a flow diagram illustrating a method of entering and exiting a standby mode in accordance with embodiments of the present invention; FIG.","23 is a block diagram of a power distribution circuit for use in relation to the CMOS imager of FIG.","1; FIG.","24 is a block diagram of a translation unit for use in relation to the CMOS imager of FIG.","1; FIG.","25 illustrates the placement of color components by the translation unit of FIG.","24; FIG.","26 is a block diagram of a flicker detection and/or correction block in accordance with embodiments of the present invention; FIG.","27 is a graph illustrating an exemplary row average across a frame of data; FIG.","28 is a flow diagram of a method for detecting and/or correcting flicker in accordance with embodiments of the present invention; FIG.","29 is a timing/block diagram illustrating the operation of qualification signals used in relation to providing parallel output data from the CMOS imager of FIG.","1; FIG.","30 is a timing diagram illustrating vertical and horizontal qualification signals used in relation to providing parallel output data from the CMOS imager of FIG.","1; FIG.","31 is a timing diagram illustrating the output of Bayer data via a parallel output interface; FIG.","32 is a timing diagram illustrating the output of REV data via a parallel output interface; FIG.","33 is a timing diagram illustrating frame dropping; FIG.","34 is a timing diagram illustrating unqualified serial output in accordance with embodiments of the present invention; and FIG.","35 is a timing diagram illustrating qualified serial output in accordance with embodiments of the present invention.","DETAILED DESCRIPTION OF THE INVENTION The present invention provides systems, devices, and methods for imaging.","More particularly, the present invention provides devices and methods for capturing images, formatting the images, and/or transmitting the images via a cellular telephone network or other wireless network.","In one particular embodiment of the present invention, an imaging device including a CMOS sensor is provided.","The imaging device includes an imager array, sensor timing control, and image processing algorithms integrated onto a single chip.","Such an imaging device can produce CIF (i.e., 352×288) resolution image for still capture, and QCIF (i.e., 176×144) resolution image for video applications.","In addition, the imaging device can perform various correction algorithms on a captured image to create a corrected YCrCb (4:2:2) image.","In an embodiment, the imager array is 384×316 physical pixels including 376×296 active pixels.","The pixels can be mapped to a configurable size of output Bayer grid array (e.g., windowing).","Pixel size can be 6.4 μm×6.4 μm.","QCIF resolution can be created by either or both of down-sampling and filtering.","In some embodiments, a resulting Bayer grid (raw RGB output) or a recreated YCrCb image is sent out on a digital ten (10) bit parallel interface.","The ten (10) bit parallel interface provides access to a captured video image on a line by line basis.","In addition, an output clock and other qualifying signals are provided in association with the ten (10) bit parallel interface.","The imaging device can include electronic exposure control from 375 nsec up to a full frame (e.g., 33.33 msec at a maximum frame rate of thirty frames per second).","Further, automatic dark level offset, a separate gain for each of the color components, and configurable offset for dynamic range stretching, as well as, image enhancement algorithms can be provided.","Such image enhancement algorithms can include black level offset, fault pixel correction, color interpolation, color and white balance correction, gamma correction, and/or auto exposure and auto white balance algorithms.","In addition, noise reduction processes can be applied to reduce noise in general, and fixed pattern noise (“FPN”) in particular.","The imaging device can operate on a single 2.8 volt power supply and a reduced-power mode can be implemented to reduce power consumption when full operation of the imaging device is not necessary.","In addition, the imaging device can include a sleep mode for further reducing power consumption.","In some embodiments, the imaging device is designed for use under the control of an external processor, which in some cases is a Digital Signal Processor.","The external processor can act as a system host (“Host”).","As such, the external processor controls the mode of the imaging device, monitors system status, and compresses an output image stream.","For example, the external processor can control the frame-rate, the size of the requested image, and several registers for image processing, such as gamma-correction and gain registers.","Such control can be provided via a programmable core integrated with the imaging device.","Block Diagram FIG.","1 illustrates a block diagram of an imaging device 100 in accordance with embodiments of the present invention.","Imaging device 100 includes a serial interface unit 106, a control unit 110, an imaging unit 130, a translation unit 150, a testability unit 170, and a parallel interface unit 190.The various units are electrically coupled via a control bus 101, an image bus 131, a translation bus 151, a parallel interface bus 191, a UART bus 120, a DeICE bus 125, and a serial bus 107.Inputs and outputs to/from imaging device 100 can be provided via an I2C input/output (“I/O”) 108, a UART I/O 121, a DeICE I/O 126, and/or a parallel video interface 192.In addition, imaging device 100 includes a clock unit 102 with a reset 171 and a clock 172 input.","Serial Interface In one embodiment, serial I/O 108 includes a clock and data input.","Such a serial interface can be used to program various registers of imaging device 100.Various different serial interface types that are known in the art can be incorporated into imaging device 100 in accordance with the present invention.","Control Unit Control unit 110 implements parameters used in relation to the different units within imaging device 100, and calculates various parameters for image enhancement.","In some embodiments, control unit 110 includes a programmable processor core 112, and in one particular embodiment, programmable processor core 112 is a S0186Turbo processor core.","Control unit 110 also includes a core memory 118, and interface hardware (not shown) to UART I/O 121 and DeICE 126 I/O.","Further, control unit 110 includes an arbitrator 116 for controlling access to control bus 101, and a frame-rate mechanism 114 for controlling the frame rate output video images from imaging device 100.Via arbitrator 116, programmable processing core 112, serial I/O 108, UART I/O 121, and/or DeICE I/O 126 can access registers maintained on imaging device 100 over control bus 101.In one particular embodiment, when programmable processing core 112 is disabled by, for example, entering a clock disable mode, serial I/O 108 is granted immediate access to control bus 101 without adhering to an arbitration scheme implemented by arbitrator 116.Such a clock disable mode is discussed in more detail with reference to clock unit 102.For purposes of the immediate discussion, clock unit 102 produces a control line that indicates if programmable processing core 112 is active, or if it is held in reset mode or has its clock shut down.","Alternatively, when programmable processing core 112 is active, arbitrator 116 grants access to control bus 101 according to a determined priority scheme.","One such priority scheme is described in the 80186Turbo reference manual.","Programmable processor core 112 serves as the main control source for imaging device 100, controlling other units via control bus 101.Such control can be provided by manipulating address, data and read/write signals associated with control bus 101.As just one example, programmable processing core 112 sets up the working mode of imaging device 100 according to the parameters that are received from a host (not shown) via serial I/O 108, UART I/O 121, and/or DeICE I/O 126.In addition, programmable processing core 112 can regulate the operation of the various units within imaging device 100.In one particular embodiment, programmable processor core 112 supports a twenty-bit address memory space for data and program memories, and a fourteen-bit address spaced for I/O mapped peripherals.","In other embodiments, only a twelve-bit memory space and an eight-bit I/O space (program and data) are supported.","Programs controlling the operation of imaging device 100 can be loaded to core memory 118 via serial I/O 108, and/or via UART I/O 121.Once the program is loaded, control unit 110 calculates and updates various image processing parameters according to a selected working mode.","After a complete frame is read from imaging unit 130 and processed by translation unit 150, control unit 110 reads statistical results of the frame, and produces the exposure, gain and auto-white balance parameters for the next frame.","In this way, control unit 110 continuously monitors operation of imaging device 100 and modifies such operation to assure that it conforms to the downloaded program.","In one particular embodiment, various parameters associated with the operation of imaging device 100 are written to registers associated with the various units within imaging device 100.Such a process can be performed by writing the parameters to core memory 118, after which programmable processing core 112 writes the values to registers maintained within the various units to which the parameters pertain.","Alternatively, such parameters can be written directly to the various units by the Host.","To facilitate such processes, imaging unit 130, translation unit 150, and parallel interface unit 190 can include double buffered registers so that parameters can be loaded into such registers at any time without interfering with continuing processes.","Imaging device 100 operates in a default standby mode, with sensor 132 turned off and translation unit 150 and parallel interface unit 190 inactive.","Once the program is loaded, control unit 110 awaits instructions from the Host.","The Host can pass commands to control unit 110 by writing to memory 118 at HOST-FIFO address space.","After writing data to the HOST-FIFO address space, the Host can activate an interrupt to programmable processing core 112.Programmable processing core 112 then reads the data and responds according to the instructions.","Once the frame counters are active, a vertical synchronization interrupt signal (“VSYNC”) is asserted every time a frame boundary for an image is detected.","VSYNC is used by programmable processing core 112 to govern software frame-rate control, or by frame-rate mechanism 114 to govern hardware frame-rate control.","When an image is being processed by imaging device 100, statistics are collected in translation unit 150.At the end of a frame processing, translation unit 150 activates an interrupt signal, END_TRANSLATION, indicating that programmable processing core 112 can read the statistics from translation unit 150.Programmable processing core 112 utilizes these statistics to calculate exposure, gain and color processing parameters for the next frame.","In some embodiments, such calculations are provided in accordance with auto-exposure, auto-white-balance, and/or dynamic range algorithms.","The calculations result in a new set of parameters that are written by control unit 110 to registers maintained in association with imaging unit 130 and translation unit 150 before the following VSYNC.","Further, control unit 110 can be interrupted by either or both of UART I/O 121 and serial I/O 108.Such interrupts are generally processed as soon as programmable processing core 112 becomes available.","Reasonable response time to such interrupts helps assure that the I/O pipelines do not become overfilled.","The following Table 1 lists a series of tasks completed as imaging device 100 is programmed and started.","TABLE 1 Imaging Device Tasks EVENT Task Comments Reset Programmable processing core 112 is frozen Deassert Reset Execute boot program - check configuration register.","If boot from UART I/O 121, wait for UART message.","If from serial I/O 108, program is loaded, and can subsequently be executed.","According to program, initiate registers and activate imager frame counters.","VSYNC Asserts Interrupt processes.","Activate translation unit 150 First frame exposure has for next frame.","Execute started, no image is being frame-rate control routine.","read out.","VSYNC Interrupt processes.","Asserts Execute frame-rate control Image is being sent from routine imaging unit 130 to translation unit 150 in accordance with a frame-rate control.","END_TRANSLATION Interrupt processes.","Asserts Read statistics results from The algorithm defines the translation unit 150.sequence of the calculations.","Calculate gain and offset for In some instances, not all imaging unit 130.Calculate calculations are performed exposure parameters, white for every frame.","balance correction, and color correction matrix.","Write calculated parameters to various units.","VSYNC Asserts Interrupt processes.","Execute frame-rate control routine.","In one embodiment, programmable processing core 112 includes six interrupts.","Such interrupts can include, but are not limited to, two software timer interrupts used for timeout purposes, and interrupts associated with UART I/O 121 and serial I/O 108.Additionally, VSYNC and END_TRANSLATION are provided.","As previously discussed, VSYNC indicates a frame boundary has been detected, and END_TRANSLATION indicates that translation unit 150 has completed processing a frame.","Upon assertion of END_TRANSLATION, translation has gathered and/or calculated various statistics associated with the processed frame that can now be read and utilized by programmable processing core 112.Programmable processing core 112 utilizes the statistics to perform one or more of the following algorithms: auto exposure (e.g., exposure parameters calculation), auto white balance (e.g., white balance coefficients calculation and/or full color matrix calculation), and dynamic range stretching, (e.g., gain and offset calculations).","Thus, for example, if the auto-exposure and auto-white-Balance options are enabled, the parameters calculated by programmable processing core 112 in relation to such controls are passed to imaging unit 130 where they are used to control exposure and processing of a subsequent image frame.","In addition, programmable processing core 112 can turn its own clock off by writing to a disable clock register maintained in clock unit 102.In some embodiments, the clock is turned back on whenever any activity is detected on a receive pin associated with UART I/O 121, or if any interrupt signal is activated.","Disabling programmable processing core 112 helps to conserve power.","In addition to controlling the rate of image output via parallel video interface 192, control unit 110 also controls the frame-rate associated with image processing internal to imaging device 100.In an embodiment, three registers are used to effectuate frame rate control.","The first, Frame_rate_en, holds an enable bit for a frame-rate mechanism 114.When frame-rate mechanism 114 is disabled, the array content is read according to a second register, Frame_rate_read, and the data is sent out on parallel interface 192.When frame-rate mechanism 114 is activated, two eight-bit registers define whether a particular frame is to be read out, or dropped.","When a frame is to be output via parallel video interface 192, a control signal, c_readframe, which is synchronized to a frame boundary signal, Im_Frame 910, is sent to imaging unit 130 to enable output of the frame data to translation unit 150.In addition, a control signal, C_Outframe, which is also synchronized to Im_Frame 910 signal is sent to parallel interface unit 190 to enable the data received from translation unit 150 to be output via parallel video interface 192 at a rate defined by the Frame_rate_send register value.","As illustrated in FIG.","2, when c_readframe is asserted high, the content of a pixel array within sensor 132 is transferred from imaging unit 130 to translation unit 150.When C_Outframe is asserted high, the data is sent out on parallel video interface 192.In some embodiments, when C_Outframe is deasserted, the output data is ignored by parallel interface unit 190, but the output pins are driven.","While the data lines of parallel video interface 192 are driven, the vertical and horizontal synchronization signals associated with the data are not driven.","In other embodiments, when C_Outframe is deasserted, the output data is ignored by parallel interface unit 190, and the output pins are not driven to conserve power and reduce noise.","Clock Unit Clock unit 102 receives clock input 172 signal and reset input 171.Based on these signals, as well as control signals from control unit 110, clock unit 102 produces the various clock and reset signals to the different units within imaging device 100.In addition, clock unit 102 samples the configuration pins while reset signal 171 is asserted, and produces several configuration signals based upon the sensed configuration arrangement.","The configuration inputs determine the working clock rate, the type of serial interface (SSI or I2C compatible) that is operational on serial I/O 108 and the program-loading source (serial interface or UART) that is operational on UART I/O 121.Further, clock unit 102 produces a control line that indicates if programmable processing core 112 is active, or if it is held in reset mode or has its clock shut down.","As previously discussed, this signal enables the bypass of arbitrator 116 for accessing control bus 101.Clock unit 102 produces the device's general reset signal.","An internal reset signal, Reset_i 104, is activated when reset input 171 is asserted, by the setting of a software reset register, or by change in phase locked loop configuration.","Clock unit 102 also synchronizes Reset_i 104 to an internal clock signal, Rclk 103.Clock unit 102 also produces a signal that resets programmable processing core 112.This reset signal is activated when Reset_i 104 is active and/or by a special reset command from serial interface unit 106.When the configuration of imaging device 100 calls for program loading through serial interface unit 106, programmable processing core 112 is held in reset state by this signal until released by a load complete command.","For power reduction purposes, in some embodiments clock signals from clock unit 102 may be turned off.","This can be accomplished under direction from either programmable processing core 112 or the Host.","As previously discussed, the clocks can be restored whenever an interrupt signal is detected.","Alternatively, clocks going to units other than control unit 110 can be turned off, and subsequently restored by issuing a command from programmable processing core 112.In an embodiment, clock unit 102 produces the following signals: an Rclk 601, an Aclk 603, a Reg_clk 609, an Rc_pix signal 605, an Rclkc, an Rclky, and an Rclkp.","Rclk 601 is the main internal clock signal generated from clock input 172.A division factor is applied to clock input 172 to produce Rclk 601, such division factor being determined by reading the configuration pins when reset input 171 is asserted.","Aclk 603 is a clock signal to the analog part of imaging unit 130 and is synchronous to Rclk 601, with a one-half Rclk 601 cycle skew between them, at the rate of the pixel-cycles.","Thus, the positive going edge of Aclk 603 follows every fourth positive edge of Rclk 601.Reg_clk 609 is a clock signal to the voltage regulators.","It is one-sixteenth the rate of Rclk 601.The rising edge of Reg_clk 609 coincides with the rising edge of Rclk 601, when Rc_pix 605 is asserted.","Rc_pix 605 is a signal in the digital domain that marks the Rclk 601 cycle that coincides with the first low cycle of Aclk 603.The first positive going clock edge after an indication that a phase lock loop is ready is qualified as Rc_pix 605.Rclkc is a clock signal to control unit 110 that is identical to Rclk 601.Similarly, Rclky is identical to Rclk 601 and is the clock signal to translation unit 150.Rclkp is a clock signal to parallel interface 190 that is derived from clock input 172.Rclkp can either be identical in frequency to clock input 172, or divided according to a clock division register, Rclkp_config.","Rclkp is used for serial output mode and is typically eight to sixteen times the rate of a pixel-clock when serial output is required.","Rclk_edge 607 is a qualifying signal to parallel interface unit 190 and is used for serial output mode.","FIG.","3 is an exemplary timing diagram 600 illustrating one example of the timing of the various signals generated by clock unit 102.The operating mode of imaging device 100 determines the clock configuration.","The following Table 2 lists the different operating modes, and the clocking scheme that is associated with each of the modes: TABLE 2 Clock Modes Input Supported Rclk divide Rclkp frequency fps Supported output format factor divide factor 8 MHz 15 fps Bayer, YUV parallel (CIF, QCIF) 1 NA Bayer 1x serial - QCIF 1 1 12 MHz 20 fps Bayer, YUV parallel (CIF, QCIF) 1 NA Bayer 1x serial - QCIF 1 1 16 MHz 30 fps Bayer, YUV parallel (CIF, QCIF) 1 NA Bayer 1x serial - QCIF 1 1 15 fps Bayer, YUV parallel (CIF, QCIF) 2 NA Bayer 1x serial - QCIF 2 2 Bayer 1x serial - CIF 2 1 YUV 1x serial - QCIF 2 1 Bayer 2x serial - QCIF 2 1 32 MHz 30 fps Bayer, YUV parallel (CIF, QCIF) 2 NA Bayer 1x serial - QCIF 2 2 Bayer 1x serial - CIF 2 1 YUV 1x serial - QCIF 2 1 Bayer 2x serial - QCIF 2 1 15 fps Bayer, YUV parallel (CIF, QCIF) 4 NA Bayer 1x serial - QCIF 4 4 Bayer 1x serial - CIF 4 2 YUV 1x serial - QCIF 4 2 YUV 1x serial - CIF 4 1 Bayer 2x serial - QCIF 4 2 Bayer 2x serial - CIF 4 1 YUV 2x serial - QCIF 4 1 64 MHz 30 fps Bayer, YUV parallel (CIF, QCIF) 4 NA Bayer 1x serial - QCIF 4 4 Bayer 1x serial - CIF 4 2 YUV 1x serial - QCIF 4 2 YUV 1x serial - CIF 4 1 Bayer 2x serial - QCIF 4 2 Bayer 2x serial - CIF 4 1 YUV 2x serial - QCIF 4 1 15 fps Bayer, YUV parallel (CIF, QCIF) 8 NA Bayer 1x serial - QCIF 8 8 Bayer 1x serial - CIF 8 4 YUV 1x serial - QCIF 8 4 YUV 1x serial - CIF 8 2 Bayer 2x serial - QCIF 8 4 Bayer 2x serial - CIF 8 2 YUV 2x serial - QCIF 8 2 YUV 2x serial - CIF 8 1 Various combinations of frequencies for the input clock, Rclk and Rclkp are summarized in Table 3 below: TABLE 3 Various Input and Output Clock Frequencies clock input 172 Rclk dividing factor Rclkp dividing factor) 8 MHz, 12 MHz, 16 MHz 1 1 16 MHz, 32 MHz 2 1 2 2 32 MHz, 64 MHz 4 1 4 2 4 4 64 MHz 8 1 8 2 8 4 8 8 Imaging Unit Imaging unit 130 provides the central functionality of imaging device 100.Imaging unit 130 includes a timing control and address generator unit (“TCAG”) 136 that receives timing signals from control unit 110 and produces the sequence signals to sensor 132 and ADC 134 for integration (e.g., exposure control), read and reset operations.","Sensor 132 includes a CIF size pixel array, level shift circuitry, gain control circuitry, programmable or black offset circuitry, gamma-correction circuitry and readout circuits.","FIG.","4 illustrates a block diagram 792 of the analog processing portion of imaging unit 130 including sensor 132 and ADC 134.Sensor 132 includes a pixel array 700, a column amplifier 703, an analog multiplexer (“MUX”) 709, a source selector mux 713, a level shift block 719, a black offset block 723, a first stage programmable gain amplifier 729, and a second stage programmable gain amplifier 733.In addition, sensor 132 includes a column logic block 739 and a reset level block 743.It should be noted that column amplifier 703 represents a vector of column amplifiers.","Column logic within imaging device 100 can be designed such that only two of the 384 total column amplifiers within column amplifier 703 are active at any given time.","FIG.","5, illustrates one embodiment of pixel array 700 associated with sensor 132.Pixel array 700 includes an array of 376 by 296 active pixels 720 surrounded by four black column is 730 on each side.","It is also surrounded by sixteen black lines 710 above and four black lines below the active image array, totaling 384 by 316.Each pixel in pixel array 700 is covered by a color filter, thus creating a Bayer grid format 800 as illustrated in FIG.","6.Referring again to FIG.","4, sensor 132 receives a number of timing and address signals from TCAG 136.More particularly, pixel array 700 receives the decoded and time-shaped line reset, Im_line_rst[328:0] (element 707), and line select, Im_line_sel[328:0] (element 727), signals.","In some embodiments of imaging device 100, each pixel within pixel array 700 includes a charge injection compensation circuit.","Referring to FIG.","7, a pixel 3000 is illustrated in accordance with embodiments of the present invention that include such a charge injection compensation circuit.","Pixel 3000 includes a reset element 3350, a source follower transistor 3360, a selection element 3370, a charge evacuation element 3380, and a light sensitive element 3390.The inputs to pixel 3000 include a reset control signal (im_line_rst 707), a selection control signal im_line_sel 727), and a charge evacuation control signal (im_line_rst_n 717).","Further, pixel 3000 provides an output signal 742.In an embodiment, pixel 3000 is attached to a voltage source 3345 and a common ground 3355.Source follower transistor 3360 can be any of a number of transistor types.","In one particular embodiment, source follower transistor 3360 is a CMOS FET.","Source follower transistor 3360 includes a source follower gate 3361, a source follower source 3362 and a source follower drain 3363.Each of evacuation element 3380, reset element 3350, and photodiode 3390 are electrically connected to source follower gate 3361.For purposes of this document, charge evacuation element 3380 can be any device capable of dissipating charge from pixel 3000.Thus, for example, charge evacuation element 3380 can be a switch, a FET, a bipolar junction transistor (“BJT”), a switched capacitor, or any other such device.","In one particular embodiment of the present invention, charge evacuation element 3380 is CMOS transistor as illustrated in FIG.","7.In some cases, such CMOS transistors can be comprised of N-FET devices, P-FET devices, and/or combinations thereof as known in the art.","In such an embodiment, charge evacuation element 3380 includes a charge evacuation gate 3381, a charge evacuation drain 3383, and a charge evacuation source 3382.As used herein, a reset element 3350 can be any device capable of switching, such that voltage source 3345 is applied to light sensitive element 3390.In one particular embodiment, reset element 3350 is a CMOS transistor that includes a reset gate 3351, a reset drain 3353, and a reset source 3352.Yet further, for purposes of this document, light sensitive element 3390 is any device that is capable of detecting the presence of light and producing a signal indicative of the amount of light detected.","Thus, in one embodiment of the present invention, light sensitive element 3390 is a photodiode.","In other embodiments, light sensitive element 3390 is a photo-gate.","Other such light sensitive elements can be used including, but not limited to, a bipolar photo-transistor and/or a stacked complex n-p-n-p device where each junction is sensitive to a part of the light spectrum to be detected.","Also for purposes of this document, selection element 3370 can be any device capable of presenting a signal representative of an amount of light impinging upon light sensitive element 3390.Thus, selection element 3370 can be a transistor, a pass gate, or the like.","In one embodiment of the invention, selection element 3370 is a CMOS transistor with a selection gate 3371, a selection drain 3373, and a selection source 3372.For discussion purposes, each of reset element 3350, source follower transistor 3360, selection element 3370, and charge evacuation element 3380 are CMOS transistors and light sensitive element 3390 is a photodiode.","However, based on the disclosure provided herein, one of ordinary skill in the art will recognize that the present invention and the principles included herewith are applicable to a number of other device types.","As illustrated by pixel 3000, source follower drain 3363 and reset drain 3353 are both electrically coupled to voltage source 3345.Further, source follower gate 3361, a node of light sensitive element 3390, reset source 3352, charge evacuation source 3382, and charge evacuation drain 3383 are electrically coupled together at a node 3303.The other node of light sensitive element 3390 is electrically coupled to column ground 3355.Yet further, source follower source 3362 is electrically coupled to selection drain 3373 at a node 3313.Reset gate 3351 is driven by im_line_rst(i) 707, charge evacuation gate 3381 is driven by im_line_rst_n(i) 717, and selection gate 3371 is driven by im_line_sel(i) 727.Output signal 742 is driven by selection source 3372.In some embodiments, im_line_rst_n(i) 717 is the inverse, or complement, of im_line_rst(i) 707.Thus, when reset element 3350 is switched, thereby providing a low impedance path from reset drain 3353 to reset source 3352, charge evacuation element 3380 is not switched, thereby providing an open circuit between charge evacuation drain 3383 and charge evacuation source 3382.The opposite is also true.","When reset element 3350 is not switched, thereby providing an open circuit between reset drain 3353 and reset source 3352, charge evacuation element 3380 is switched thereby providing a low impedance path from charge evacuation drain 3383 to charge evacuation source 3382.In some embodiments where im_line_rst_n(i) 717 is the complement of im_line_rst(i) 707, im_line rst_n(i) 717 is delayed such that a falling edge of im_line rst(i) 707 precedes a corresponding rising edge of im_line_rst_n(i) 717 by a period of time.","In other embodiments, im_line rst_n(i) 717 works in relation to im_line rst(i) 707, but is not the complement of im_line_rst(i) 707, but rather a distinctly generated signal.","The timing relationships of the various signals are discussed further in relation to FIG.","9 below.","FIG.","8 illustrates a flow diagram 3400 of one method in accordance with the present invention for operating pixel 3000.In operation, im_line_rst(i) 707 is asserted such that a low impedance path exists between reset drain 3353 and reset source 3352 (block 3410).","In particular embodiments, im_line_rst(i) 707 overdrives reset gate 3351 such that photodiode 3390 is charged as rapidly as possible.","Further, im_line_rst_n(i) 717 is deasserted such that an open circuit exists between charge evacuation drain 3383 and charge evacuation source 3382 (block 3420).","As such, a voltage corresponding to voltage source 3345, less the impedance drop across reset element 3350 is present at node 3303, thereby reverse biasing photodiode 3390.In this state, photodiode 3390 is charged to a level corresponding to the voltage at node 3303.Further, a negative charge is built in the channel of reset element 3350.Some time later, im_line_rst(i) 707 is deasserted such that an open circuit exists between reset drain 3353 and reset source 3352 (block 3430).","Thus, additional charge from voltage source 3345 is not available to photodiode 3390.However, on the falling edge of im_line_rst(i) 707, the negative charge accumulated in the channel of reset element 3350 is dissipated through both reset drain 3353 and reset source 3352.Thus, part of the charge is discharged via node 3303 causing a voltage drop (i.e., noise), which can be significant.","In some cases, the voltage drop due to the negative charge from reset element 3350 can be between two-hundred and five-hundred millivolts for geometries of four to seven micrometers, and is exacerbated as pixel geometries and source voltages decrease.","With an open circuit between reset drain 3353 and reset-source 3352, the charge built up in photodiode 3390 begins to dissipate at a rate corresponding to the amount of light impinging upon photodiode 3390.Thus, where a significant amount of light impinges upon photodiode 3390, the voltage at node 3303 will decrease at a more rapid rate than if a small amount of light impinges upon photodiode 3390.Either some time after the deassertion of im_line_rst(i) 707 or coincident therewith, im_line_rst_n(i) 717 is asserted such that a low impedance path exists between charge evacuation drain 3383 and charge evacuation source 3352 (block 3440).","This assertion provides a path whereby at least a portion of the negative charge that was stored in the channel of reset element 3350 can dissipate through charge evacuation device 3380.In some embodiments, the type of device used to implement charge evacuation element 3380 is chosen such that the operation of charge evacuation element 3380 reflects the non-linearities exhibited by reset element 3350.Thus, in some cases, charge evacuation element 3350 and reset element 3350 are chosen to be the same device types.","As approximately half of the charge built up in the channel of reset element 3350 dissipates through reset drain 3353 and the other half dissipates through reset source 3352, the size of charge evacuation device 3380 can be chosen such that it can evacuate roughly half of the charge built up in the channel of reset element 3350.Thus, in one embodiment, where the charge evacuation element 3380 and reset element 3350 are the same device type, the size of charge evacuation device 3380 is roughly half the size of reset element 3350.In other embodiments where reset element 3350 and charge evacuation element 3380 are different device types, such as, when charge evacuation element 3380 is a switched capacitor, the size of the capacitor is chosen based on the amount of charge to be dissipated from the channel of reset element 3350 via reset source 3352.As illustrated below in FIG.","9, dissipation of the negative charge in the channel of reset element 3350 through charge evacuation element 3380 can, depending upon the geometry of pixel 3000, eliminate as much as five-hundred millivolts of noise at node 3303.Elimination of such noise at node 3303 assures that the signal sampled at output 742 more accurately reflects the amount of light detected at photodiode 3390.At some point, im_line_sel(i) 727 is asserted to sample the remaining voltage at node 3303, and thereby gain an approximation of the light impinging upon photodiode 3390 (block 3450).","With im_line_sel(i) 727 asserted, a low impedance path exists from selection drain 3373 to selection source 3372.Thus, a signal representative of the voltage present at node 3303 is presented on output 742 via source follower transistor 3360.Once the output is sampled, im_line_sel(i) 727 is deasserted (block 3460) and the process is repeated to detect the amount of light impinging upon photodiode 3390 at a future point in time.","It should be recognized from FIG.","9 that in some embodiments of the present invention, the assertion of im_line_sel(i) 727 overlaps the assertion of im_line_rst(i) 707.Yet, in other embodiments, the assertion of im_line sel(i) 727 does not overlap the assertion of im_line_rst(i) 707.FIG.","9 illustrate timing diagrams of the various signals related to the operation of pixel 3000.More particularly, the figures illustrate the relationship between im_line_rst(i) 707, im_line rst_n(i) 717, im_line_sel(i) 727, the voltage at node 3303, and the voltage at output 742.FIG.","9a is a timing diagram 3501 illustrating operation of pixel 3000 where im_line_rst(i) 707 and im_line_rst_n(i) 717 are complementary, and where charge evacuation element 3380 is operational.","In contrast, FIG.","9b is a timing diagram 3502 illustrating similar operation of pixel 3000 where charge evacuation element 3380 is not present.","As illustrated in FIG.","9b, at the falling edge of im_line_rst(i) 707 (noted as 3520), a voltage drop, VDROP 3510, is exhibited at node 3303.This voltage drop exists until photodiode 3390 is again charged by assertion of im_line_rst(i) 707 (noted as 3530).","This voltage reduction at node 3303 (i.e., noise) causes a corresponding reduction in output signal 742.This is in comparison to timing diagram 3501 of FIG.","9a, where the voltage drop at node 3303 does not occur because the charge causing the voltage drop in timing diagram 3502 is evacuated through charge evacuation element 3380 as previously described.","Thus, as illustrated in the contrast between FIG.","9a and FIG.","9b, a substantial amount of noise can be eliminated from node 3303 via the operation of charge evacuation element 3380.FIG.","9c is a timing diagram 3503 that illustrates another embodiment where, similar to timing diagram 3501, im_line_rst(i) 707 and im_line_rst_n(i) 717 are complementary.","However, in contrast to timing diagram 3501, timing diagram 3503 illustrates im_line_rst_n(i) 717 delayed from im_line_rst(i) 707 by a period 3570.As illustrated, charge evacuation element 3380 still dissipates the charge from the channel of reset element 3350, albeit at a slightly later time corresponding to the assertion of im_line_rst_n(i) 717.Thus, a voltage droop 3580 corresponding to VDROP 3510 (illustrated as a dotted line) is noted on node 3303, until the assertion of im_line_rst_n(i) 717 (noted as 3590).","However, because of the operation of charge evacuation element 3380, the voltage at node 3303 is corrected before the sample is taken upon assertion of im_line_sel(i) 727.Based on the disclosure provided herein, one of ordinary skill in the art will recognize a number of other timing relationships between the various signals that each dissipate the charge within the channel of reset element 3350, thus reducing the noise manifest at output 742.Further, as previously mentioned, im_line_rst(i) 707 and im_line_rst_n(i) 717 can be separately generated to achieve a timing relationship that allows for charging of photodiode 3390, and for dissipating the charge within the channel of reset element 3350.In some cases, it can minimize the logic in TCAG 136 to have im_line_rst_n(i) 717 be the complement of im_line_rst(i) 707.Further, it should be recognized that im_line_sel(i) 727 can be generated at other times than those illustrated in FIG.","9.FIG.","10 illustrates the architecture of column amplifier 703.Discharge (element 777) and enable (element 767) signals are received from column logic block 739.Using column amplifier 703, the output of each column is double sampled by the track and hold circuits in the column amplifier that are controlled by im_smp_strobe (element 747) and im_ref_strobe (element 757) signals.","A timing diagram 893 illustrating the inter-relationship of the various signals of pixel array 700 and column amplifier 703 is provided as FIG.","11.Of note, the exposure control described in more detail below should not have a line in pixel array 700 during a period 892 outlined by dotted lines on FIG.","11.Imaging unit 130 can be programmed to provide a standard output from pixel array 700 or a mirrored image.","FIG.","12 provides a timing diagram of the signals associated with outputting the image in standard mode, and FIG.","13 provides a timing diagram of the signals associated with outputting the image in mirrored mode.","Selection between the standard and mirrored modes is accomplished by programming an im_mirror (element 787) signal via a register bit, which drives column logic block 739.Referring again to FIG.","4, analog mux 709 selects the pixel from pixel array 700 that will be processed.","Thus, a column of pixel array 700 is selected by column logic block 739, and then a single pixel within the column is selected via analog mux 709.Analog mux 709 is a three level mux where the first and second levels are eight to one muxes, and the third level is a six to one mux.","This provides for a total of a 384 to one mux.","Selection is accomplished via column address signals (element 797) im_col_add_L[7:0] for selecting the first level mux im_col_add_M[7:0] for selecting the second level mux, im_col_add H[5:0] for selecting the third level mux.","Sensor 132 can operate in a rolling mode, in which the exposure and readout processes are linked.","The sampling of each row data is performed at the end of the integration time.","After sampling a selected-row data, the row is reset.","The row is sampled again after the reset.","This second sampling helps eliminate fixed pattern noise (FPN) by implementing a correlated double-sampling procedure.","The integration of that row starts again according to the exposure parameters.","A set of registers defines the total size of pixel array 700 and the size of the array that is read out and sent to translation unit 150 for further processing.","Another set of registers defines the exposure time that is required for pixels within pixel array 700.The exposure and read timing sequences are derived from these registers.","The processing of a single pixel of sensor 132 is referred to as a pixel cycle.","There are four Rclk 601 cycles in a pixel-cycle that is controlled by a timing circuit.","An Im_Pix signal from clock unit 102 qualifies the first clock edge of the pixel-cycle.","On activation of the timing circuit, line and pixel counters are set to zero, and subsequently begin counting pixel-cycles.","An Im_Frame 910 signal is activated on the first pixel-cycle of the frame, and an Im_Line 920 signal is activated on the first pixel-cycle of a line, except for the first line, when Im_Frame 910 is active.","The pixel counter counts upon activation of the Im_Pix signal, and the line counter is activated when the pixel counter reaches a full-line count.","On activation of Im_Line 920 after activation of Im_Frame 910, TCAG 136 reads and resets the first line in pixel array 700.On each activation of Im_Line 920, the lines are only reset.","Once the line whose index is equal to the Im_starty register value is reached, data is sent out starting at the pixel whose index matches the Im_startx register value.","The pixels that are inside the window defined by the Im_starty and Im_startx, Im_endy, and Im_endx converted from the analog to digital domain using ADC 134, and passed to translation unit 150.An additional window can be defined for differentiating between pixels in “area of interest”, and pixels outside this defined area.","An Im_window signal is active for those pixels inside the window, and inactive for the pixels outside the window.","Im_Line 920 starts the readout process of an image line, and also causes a reset to that line.","One or more register values define the reset time for the first line.","Further, a coarse exposure register value defines the number of lines between Im_Frame 910 signal activation and the end of the coarse reset time.","Once the appropriate Im_Line 920 is reached, the number of pixel-cycles is counted according to a fine exposure register value.","Once the reset time has elapsed, the integration time of the first line is initiated.","The event is marked by the activation of Im_Expline 930 for one pixel-cycle.","A second nine-bit counter is then loaded with the full-line length register and decremented on each pixel cycle.","Once the nine-bit counter reaches a zero value, the integration time of the second line is initiated.","Im_Expline 930 is activated for one pixel-cycle and the process repeats itself until the integration time of the last line is initiated.","FIG.","14 illustrates the interaction of the previously described frame timing signals.","Each activation of Im_Expline 930 starts the exposure of an image line, and In_Line 920 signal starts the readout and reset processes.","As illustrated in FIG.","15, the line is sampled twice, once at the end of the exposure time (element 1010), and for the second time right after the activation of the reset signal (element 1020).","Double buffering can be used for all frame parameters.","This allows the Host or programmable processing core 112 to write the next frame parameters at any time.","Implementation of the newly written parameters are, however, delayed until activation of the following Im_Frame 910 to avoid problems with the currently processing frame.","Referring again to FIG.","4, a signal created by a selected pixel within pixel array 700 is passed through analog mux 703 and source selection mux 709 prior to further processing by sensor 132.More particularly, the selected pixel signal is level shifted by level shift block 719, black level corrected by black offset block 723, and amplified by first stage programmable gain amplifier 729, and second stage programmable gain amplifier 733.The output of the amplified and corrected pixel signal is then passed from sensor 132 to ADC 134.In one particular embodiment of imaging device 100, the level shift, and black offset are done in the digital domain.","In other embodiments, the processes are performed in the analog domain, while in yet other embodiments, the level shift and amplification are performed in the analog domain, while the black offset is performed in the digital domain.","Thus, it should be recognized that the point at which a selected pixel signal is presented to ADC 134 can be changed depending upon the particular embodiment.","Further, the order in which amplification, black offset and level shifting are performed can be modified.","For example, the selected pixel signal can be level shifted and amplified in the analog domain, passed to ADC 134, and subsequently black level corrected by black offset block 723 in the digital domain.","As further discussed below, analog signal processing portions of imaging device 100 are designed to ensure that as much as possible of an image detected by pixel array 700 is passed from the analog domain to the digital domain.","Further, power and bandwidth considerations are entertained in deciding whether the processing is performed in the analog or digital domain.","Thus, the order of processing through ADC 134, level shift block 719, black offset block 723, amplifiers 729 and 733, as well as whether the blocks are implemented in the analog or digital domain can be adjusted to ensure that the entire resolution of ADC 134 is exploited, adjusted based upon power and bandwidth considerations, and/or a combination thereof.","As used herein, dark level is the level of the pixel signal due to leakage and other circuit related effects that occurs in unexposed pixel during a given exposure time.","The dark level can be obtained by reading the pixels of the dark rows.","Black level is the level of the pixel signal corresponding to the darkest pixel in a frame (e.g., shadowed areas of an image).","White level is the level of the pixel signal corresponding to the brightest pixel in a frame (e.g., highlighted areas of an image).","As illustrated in FIG.","4, the selected pixel signal passed to ADC 134 is described by the equation: V adc = { ( V max - Δ V ref ) * ( 1 + im_lsh _gain ) - ( - 1 ) im_black [ 4 ] Δ V ref im_black [ 3 : 0 ] 16 } * 2 ( 1 + im_gain _m [ 2 : 0 ] ) 1 + im_gain _d [ 1 : 0 ] The signals in the analog processing chain are differential voltages in the range negative one volt to positive one volt.","The uncorrected and un-amplified output from source selector mux 713 is in the range of zero volts to one and one-half volts.","Level shift block 719 converts the output from source selector mux 713 to a voltage range compatible with the other analog processing circuits.","In some embodiments, level shift block 719 is operable in one of two different modes: a high gain mode and a high dynamic range mode.","The high gain mode is selected by setting a register output im_lsh_gain 714 equal to a high level.","The high gain mode operates by mapping a zero to one volt input range to a negative one volt to positive one volt range.","Alternatively, the high dynamic range mode is selected by setting a register output im_lsh_gain 714 equal to a low level.","The high dynamic range mode operates by mapping a zero to two volt input range to the negative one volt to positive one volt range.","FIG.","16 illustrates the correction and amplification of the selected pixel signal as it is received from source select mux 713 and passed through level shift block 719, black offset block 723, and amplifier stages 729, 733 for both high gain mode (signal 844) and high dynamic range mode (signal 846).","For a given exposure time, the voltages of most pixels in the frame will be in the range between the black level and the white level.","In some embodiments, ADC 134 ideally converts only this voltage range, without losing resolution by eliminating signals outside of the range.","In one particular embodiment, adjusting the voltages of the pixels to be in the range between black level and white level is achieved by black offset block 723.To this end, a control signal, im_black[4:0] (element 724), can be provided in sign-magnitude format and use to program the level adjustment to the proper level.","In one embodiment, the resolution of black offset block 723 is equivalent to the thirty-two least significant bits of ADC 134.FIG.","17 illustrates examples of signal outputs for normally lighted (signal 853), dark (signal 857), and bright (signal 859) subjects from source select mux 713 that are passed through level shift block 719, black offset block 723, and amplifier stages 729, 733.The gain and black level values are all pixel color dependent.","The red-green and blue-green ratios of the gain settings are determined in order to achieve gross white balance.","An exemplary timing of the control signals for the analog processing circuits is presented in FIG.","18.A sel[1:0] (element 774) is representative of the timing of column address signals 797 and also and im_test[7:0] (element 704) signal.","In one particular embodiment, a maximum exposure time is one frame at thirty frames per second, or 33.33 milliseconds, and a maximum gain of the analog chain is thirty-two (i.e, two from level shift block 719, and four from each of amplifiers 729 and 733).","A pixel sensitivity of 0.226 [V/lux*sec] for a green pixel at 540 nm is provided by pixels within pixel array 700.Full scale of ADC 134 is two volts (e.g., negative one volt to positive one volt differential), where the least significant bit is 1.95 mV.","Using these parameters, the size of the analog processing section of imaging device 100 can be determined and optimized.","For determining the resolution used to obtain the proper exposure, the lowest illumination level that can be translated to full scale response of ADC 134 corresponds to the maximal exposure time and maximal amplifier gain.","The following equation describes the approach: 2[V]/(32*0.0333[sec]*0.226[V/lux*sec])=8.3[lux] If the highest level of illumination is 100,000 [lux], the lowest exposure time needed assuming minimal amplifier gain of unity is: 2[V]/(1*100,000[lux]*0.226[V/lux*sec])=88.5 [usec] As in a CIF frame there are 101,376 pixels, the minimal exposure time corresponds to about three-hundred, fifty-four pixel cycles at thirty frames per second.","Therefore, controlling exposure time at the resolution of one pixel cycle is adequate.","For sizing black offset block 723, it is assumed that the exposure is set so that at the output source select mux 713, the white level corresponds to a drop of one volt and the black level corresponds to a drop of 937.5 mV.","If the amplifier 729, 733 gain is set to sixteen, and the black level DAC is five-bit (sign magnitude) with the least significant bit equaling 62.5 mV is set to fifteen, this will stretch the narrow histogram on the full resolution of ADC 134.On the opposite end, for a dark picture where at the output of source select mux 713 the black level is zero and the white level corresponds to a drop of 62.5 mV, a setting of negative fifteen for the black level DAC and a gain of sixteen for the amplifier 729, 733 will stretch the narrow histogram on the full resolution of ADC 134.The programmable gain amplifier 729, 733 has a maximum gain of sixteen and supports several functions including extending the exposure range for low illumination levels, providing gross white balance, and stretching the histogram as described in relation to black level correction.","Programmable gain amplifier 729, 733 can be implemented as a two stage amplifier, where each stage exhibits a maximum gain of four.","First stage programmable gain amplifier 729 is described by the following equation: Gain PGA_D = 4 1 + im_gain _d [ 1 : 0 ] Second stage programmable gain amplifier 733 is described by the following equation: Gain PGA_M = 1 + im_gain _m [ 2 : 0 ] 2 There is separate gain control for each color filtered pixel.","The parameters associated with the gain control are used for coarse white balance, and for enhanced exposure in dim light situations, where the maximum exposure time has been reached, but the output image is still too dark.","The gain parameters are used for expanding the range of the sampled data to the full range of ADC 134.The first sixteen lines of pixel array 700 are dark lines that are not exposed to light.","Each of the lines are integrated similar to other lines and the dark level for each color is separately measured on the output of ADC 134.The average of the measured dark line values is passed back for Dark pixel correction of the lines that follow the dark lines in pixel array 700.The effect can be immediate, where the calculation result is activated during the readout of the same frame where the dark lines were evaluated.","Image device 100 includes two methods for calculating the dark offset.","One or the other mode is selected by writing a predetermined mode selection in a register.","The two modes are dark offset per column and average dark offset.","Dark offset per column involves calculating and storing an offset value for each column.","When reading the image area, the dark offset parameter selected based on the particular column that is currently being read from pixel array 700, where the line number indicates the color being read.","The offset for each column is averaged on the fifth to twelfth lines, giving a total of eight lines.","The offset for each color component in each column is c averages of four pixels of the same column and color, minus the fixed black offset that is applied during these lines.","During the dark offset calculation, the black offset inputs are set to a positive known value and stored in a register, the color dependent gain is applied, and the output of ADC 134 is sampled.","Two RAMs of 384 by twelve-bits can be used.","The output from ADC 134 for the first and second dark lines is written to the RAMs.","For the following lines, the even-numbered line values are added to the first RAM entries, and the odd-numbered line values to the second RAM entries.","After the seventh line (a “RED” line), values are added and the result is stored back in the RAM.","The next line is processed in the same manner for the “BLUE” line offset.","Throughout this process, the average offset per color is calculated as well.","During the period for lines twelve and thirteen, which are dark lines that are not read from the array, the RAM contents are read, and the “column noise” is calculated as follows: Column—offset=(RAMsum)/4, rounded up Noise=Column—offset−Average, 12 bits two's complement.","When reading the image pixels from pixel array 700, the “noise” dark offset is read from the RAM.","The correction is performed in the digital domain on the output of ADC 134.The values that are stored in the RAM are subtracted from the output of ADC 134 according to the column number and the line type that is being read.","To simplify calculations, average dark offset mode is utilized.","In the average dark offset mode, the first four lines are skipped, and only the next eight dark lines are evaluated.","Evaluation is performed from pixel number sixty-four to pixel number three hundred nineteen, totaling 256 pixels per line.","A total of 128×2 green, 128 blue and 128 red pixel values are collected.","The result is averaged, limited, and stored in registers.","Further, in some embodiments, three four-bit black offset parameters are stored in a register for each of the three colors.","The value in the registers are passed to imaging unit 130 and subtracted from or added to the analog image signal before the signal is passed to ADC 134.Thus, three four-bit digital to analog converters (“DAC”) are used to convert the register values to the analog domain before the subtraction process is accomplished.","The operation is performed in accordance with a sign bit that is passed along with each of the three register values.","In some instances, the four-bit offset values are each multiplied by sixteen before the subtraction process is completed.","In addition, some embodiments of imaging device 100 include provisions for reducing random noise.","Such provisions include, but are not limited to, applying the aforementioned charge injection technique to pixel reset signals, stopping operation of the various digital circuitry in imaging device 100 while each row of pixel array 700 is sampled, and providing a clock driving the analog circuitry that precedes the clock driving the digital circuitry to keep various substrate noise effects orthogonal in time domain.","In one embodiment, such a phase difference between the analog clock and the digital clock is provided by deriving the analog clock from the falling edge of the digital clock, and subsequently adding some delay to the analog clock before applying it to ADC 134.Some embodiments include additional measures to eliminate random noise that include, but are not limited to, providing analog processing switched capacitor circuits that use clock phases generated by ADC 134 and synchronizing control signals driving the analog circuitry using the clock generated by ADC 134.In addition, the sizes of the capacitors used in the switched capacitor circuits can be increased in order to meet the precision requirements and to reduced kT/Cnoise.","Some embodiments of the present invention provide for reducing power consumption of imaging device 100.Such processes can include reducing power consumption of the analog processing circuitry of imaging device 100, while in other embodiments, power consumption is reduced by reducing power consumption of the digital processing circuitry of imaging device 100.In one particular embodiment, power consumption is reduced by limiting power consumption by both the analog processing circuitry and the digital processing circuitry of imaging device 100.FIG.","19 is a timing diagram illustrating the relationship between power consumption control signals, Im_stdby 764 and im_adc_cnt[4] 754.As previously illustrated in FIG.","4, Im_stdby 764 is provided to the various analog processing circuits of sensor 132 including analog mux 709, source selection mux 713, level shift block 719, black offset block 723, and programmable gain amplifier 729, 733.Im_adc_cnt[4] 754 is provided to ADC 134.Following FIG.","19, when Im_stdby 764 is asserted high (noted 1920), the analog processing circuitry stops functioning during a non-functional period 1910 surrounded by dashed lines.","One cycle of aclk 603 after assertion of Im_stdby 764, Im_adc_cnt[4] 754 is asserted high (noted 1940), thus placing ADC 134 in the standby mode.","To awake from the standby mode, Im_adc_cnt[4] 754 is deasserted (noted 1950) and twenty cycles of aclk 603 later (noted 1960), Im_stdby 764 is deasserted (noted 1970).","Once Im_stdby 764 is deasserted, the analog processing circuitry becomes operational again one cycle of aclk 603 later (noted 1980).","During non-functional period 1910, the analog processing circuits will enter a low power consumption standby mode.","In one embodiment of the standby mode, the common mode feedback of the operational amplifiers remains operational, but the circuits associated with the various analog processing calculations are disabled.","This maintains the inertial state of the amplifiers, while reducing the power consumption of the analog circuitry associated with imaging device 100.FIG.","20 illustrates a block diagram 1905 of circuitry related to controlling the standby mode.","Block diagram 1905 includes pixel array 700 providing differential inputs 1903 to a processing control system 1925.Processing control system 1925 provides either differential inputs 1903 to an analog processing circuit 1955 via differential inputs 1906, or a differential reference voltage 1915 (received via inputs 1904).","Differential inputs 1906 are processed by analog processing circuitry 1955 as previously discussed, and the results presented via differential inputs 1907 to ADC 134 are converted to the digital domain and passed to a digital processing circuit 1975 via a bus 1965.Digital processing circuit 1975 performs the various digital processing functions as previously discussed, and the processed data is provided as an image output.","In some embodiments analog processing circuit 1955 includes column amplifier 703, analog mux 709, source selection mux 713, level shift block 719, black offset block 723, and programmable gain amplifier 729, 733.In other embodiments, analog processing circuit 1955 includes a subset of the aforementioned element.","For example, in one particular embodiment, analog processing circuit 1955 includes level shift block 719, black offset block 723, and programmable gain amplifier 729, 733.Thus, depending upon the design, one or more of the aforementioned blocks can be placed in standby mode either individually, or as a group.","Further, in some embodiments, digital processing circuit 1955 includes translation unit 150 and parallel interface unit 190.In other embodiments, digital processing circuit 1955 includes only one or the other of translation unit 150 and parallel interface unit 190.In some cases, only a subset of the pixels within pixel array 700 are processed and provided as an image output.","For example, bandwidth available for the image output may be less than that required to output each pixel of pixel array 700.As a more specific example, every other pixel line and/or pixel column of pixel array 700 may be dropped.","In such cases, embodiments of the present invention provide for limiting power consumption of imaging device 100 by placing analog processing circuit and/or digital processing circuit 1975 in the standby mode.","To do this, a sub-sample control system 1945 provides an indication about whether a particular line, column and/or pixel is to be processed.","This indication From sub-sample control system 1945 is used to generate Im_stdby signal 764 which, as previously discussed, controls whether analog processing circuit 1925 and/or digital processing circuit 1975 are to enter the standby mode, or remain operational.","In some embodiments, sub-sample control system 1945 is a hardware system utilizing information from frame rate block 114 of control unit 110 and/or TCAG 136 of imaging unit 130 to determine where in a frame a received pixel is from, and whether the pixel is to be processed.","Alternatively, in other embodiments, sub-sample control system 1945 is implemented in software and executed by programmable processing core 112.In yet another embodiment, sub-sample control system 1945 is a hybrid hardware and software implementation utilizing relevant portions of TCAG 136, frame rate block 114, and/or programmable processing core 112.In one particular example where only two of every four pixel lines are processed, sub-sample control system 1945 monitors the addressing of pixel array 700 and causes Im_stdby 764 to assert each time a non-processed pixel line is addressed.","This causes processing control system 1925 to present reference voltage 1915 to analog processing circuit 1955 in place of differential input 1903 from pixel array 700.Reference voltage 1915 is electrically coupled to switching inputs of analog processing circuit 1955, thus reducing power dissipation due to switching circuits in analog processing circuit 1955.However, the common mode feedback of the operational amplifiers within analog processing circuit 1955 continue operating.","This maintains the inertial state of the amplifiers, while reducing the power consumption of the analog circuitry associated with imaging device 100.In addition, Im_adc_cnt[4] 754 is asserted to place ADC 134 in a similar standby mode.","Yet further, a digital disable signal 1985 is provided from processing control system 1925 to digital processing circuit 1975.This signal is used to gate the various clocks distributed to the various operating units of imaging device 100.In some embodiments, processing control system 1925 comprises portions of Rclk unit 102.In such embodiments, the various clock signals are gated by Rclk unit 102 and there is no reason to provide digital disable signal 1985 to digital processing circuit 1975.FIG.","21a illustrates all exemplary circuit 1943 used to electrically couple analog circuits within analog processing circuit 1955 to reference voltage 1904.In circuit 1943, PH1 and PH2 are non-overlapping clock phases.","In standby mode, there is no switching to vref 1904a and vref_n 1904b, rather inputs 1903 are shorted to vcommon 1953 to limit switching.","With such a circuit, both the bias currents of the Op-Amps, as well as the switching related to analog calculations is reduced.","However, switching sufficient to keep the common mode feedback loops of an amplifier 1963 active is maintained.","When Im_stdby 764 is asserted high, standby switches 1964 are shut and operational switches 1965 are open.","Thus, switching in circuit 1943 is limited.","FIG.","21b illustrates exemplary circuit 1943 when Im_stdby 764 is deasserted, and circuit 1943 is operating in phase one, PH1.FIG.","21c illustrates exemplary circuit 1943 when Im_stdby 764 is similarly deasserted, but circuit 1943 is operating in phase 2, PH2.FIG.","22 is a flow diagram 1974 illustrating a method according to the present invention for entering and exiting the standby mode.","Following flow diagram 1974, parameters associated with sub-sample control system 1945 are set-up (block 1976).","This can include writing various registers controlling the portions of pixel array 700 that are processed.","Such a register write can be performed by programmable processor control 112 and/or the Host.","An image is detected from pixel array 700 (block 1977).","Pixel array 700 is addressed by TCAG 136 to cause a desired column of pixel signals to be presented by pixel array 700 (block 1978).","If sub-sample control system 1945 indicates that the column is to be processed (block 1979) and that the line is to be processed (block 1981), then the analog processing circuitry 1955 is enabled, or awakened from the standby mode (block 1982).","Further, digital processing circuitry 1975 is enabled by removing the clock gate (block 1983).","With the circuitry thus enabled, the pixel signals received from pixel array 700 are processed.","Alternatively, where either the pixel column (block 1979) or the pixel line (block 1981) is not to be processed, analog processing circuit 1955 (block 1986) and digital processing circuit (block 1987) are placed in the standby mode to conserve power.","In such a state, the pixel signals received from pixel array 700 are dropped and not processed (block 1988).","The power distribution and management scheme for the analog circuitry provides a supply of 2.8 volts +/−10% supply.","On-chip regulators are used to provide the following supply voltages: —2.5 volts +/−10% for the 0.25 u digital circuits, 2.4 volts +10/−5% for the imager array on a vddArr 881 supply, and 3.2 volts +10/−5% for the imager array line reset drivers.","A block diagram 896 of the power management for imaging device 100 is illustrated in FIG.","23.As illustrated, the regulators are controlled by the following signals: a reg_d enable 882 signal the when asserted low causes the digital regulator to enter standby mode with the 2.5 V voltage is supplied from an external source.","In order to enable power up, reg_d_enable 882 is gated with an atest 883 signal as shown in FIG.","23.At power up atest 883 is kept low forcing reg_d_enable 882 high.","When reg_d_enable is 882 is high, the digital regulator output voltage is reduced to 2.0 V. The order of power-up of the regulators has to be digital regulator 884 first and only then a boost regulator 885.The clock of boost regulator 885 has to be provided by the digital circuits.","It is a clock in the range of 500 KHz to 1 MHz, obtained by dividing Rclk 601 by sixteen.","The edges of the divided clock should be aligned with the digital signals (i.e., aligned with the rising edge of Rclk 601).","To facilitate testing and/or characterization, imaging device 100 includes one or more built in self test (“BIST”) routines.","In particular embodiments, the BIST routines provide the ability to test one or more of blocks 700, 703, 709, 713, 719, 723, 729, 733, and/or 134 using a digital tester.","In some cases, such testing can further be accomplished with or without illumination of imaging device 100.Such approaches can reduce the costs and increase the rate at which the functionality of imaging device 100 is verified.","In some embodiments, one or more additional lines of “pseudo”-pixels are included within pixel array 700.The output from the pseudo-pixels can be a pre-defined voltage that remains constant regardless of any light impinging upon pixel array 700.For example, in one instance, the pseudo-pixels have an output of one volt that is passed to the column amplifiers.","These pixels are activated by asserting im_line_test (element 737).","The outputs of the pseudo-pixels are passed through column amplifier 703, and analog mux 709.In turn, the outputs are passed though source selector mux 713 by proper setting of im_test[8:0] (element 704).","Im_test[8:0] (element 704) is derived from decoding a programmable digital register (im_test_en[3:0]).","The outputs are then passed through one or more of blocks 719, 723, 729, 733, and 134.The output of ADC 134 can then be compared against a range of values to determine if imaging device 100 is within acceptable design limits.","The comparison can be performed either by a programmable hardware comparator, a hardwired hardware comparator, or via programmable processor core 112.Such a test provides an ability to test the operation of column amplifier 703, and/or one or more of blocks 719, 723, 729, 733, and 134.As the inputs and outputs associated with the test are available in the digital domain, a purely digital test of the analog circuit can be performed.","This provides an efficient and cost effective means for verifying the functionality of imaging device 100 as testing of the analog circuitry can be performed on a digital tester.","As previously stated, an output to be tested using one or more of the BIST routines can be passed through one or more of blocks 719, 723, 729, 733, and 134.Which blocks the outputs pass through is programmable.","Using such a programmable approach, each block can be individually bypassed.","Such bypassing allows for the output of each block individually, or a combination of the blocks to be tested to be isolated for testing purposes.","Bypassing of the individual blocks is controlled by signals im_by_lsh 702 to bypass level shift block 719, im_by_blk 712 to bypass black offset block 723, im_by_pgd 722 to bypass first stage programmable gain amplifier 729, and im_by_pgm 732 to bypass second stage programmable gain amplifier 733.Further, in some embodiments, source selector mux 713 permits application of various test voltages to the input of the analog processing circuitry.","This provides for applying known analog voltages to one or more portions of analog processing blocks 719, 723, 729, 733, 134.In one embodiment, the digital output generated by applying the known analog voltage to the blocks is compared against a predetermined value, or range of values.","In this way, testing of analog blocks 719, 723, 729, 733, 134 can be performed as a purely digital test, where im_test (8:0) (element 704) is generated from a programmable digital register (im_test_en[3:0]), and the outputs are each tested in the digital domain.","Again, this provides an efficient and cost effective means for verifying the functionality of imaging device 100 as analog testing can be performed on a digital tester.","Further, as each of blocks 719, 723, 729, 733 can be individually bypassed, any combination of blocks 719, 723, 729, 733 can be verified using the digital test.","In one particular embodiment of the present invention, source selector mux 713 selects between five input voltages based on im_test(8:0) (element 704).","More particularly, source selector mux 713 permits connecting reference voltages (v_ref 706, common 716, and vref_n 726) that are generated internal to imaging device 100 to the input of the analog processing blocks.","In some embodiments vref 706 and vref_n 726 are derived from ADC 134, and common 716 is a provided via a resistive divider coupled to the source voltage.","In addition, source selector mux 713 also provides for selecting externally generated reference voltages, atest 736 and atest_n 746 that can be bonded to input pads of imaging device 100.By selecting one of the five reference voltages 706, 716, 726, 736, 746, a known voltage can be applied to one or more of blocks 719, 723, 729, 733, and output via ADC 134.The output of ADC 134 can then be compared against a range of values to determine if imaging device 100 is within acceptable design limits.","In some embodiments, an additional test mechanism provides for digital testing of each column within pixel array 700.In such a test, pixel outputs from pixel array 700 are paired with an output from reset level 743.Reset level 743 consists of a group of pixels kept under a reset state and a column amplifier identical to column amplifier 703.BIST pixels within pixel array 700 are selected and the outputs from the pixels are passed through a selected portion of column amplifier 703 and analog mux 709.One side of the output is then paired with a known reset level 743 signal and selected via source selector mux 713.This pairing between the known reset level 743 and the output from the various BIST pixels is then passed through one or more of blocks 719, 723, 729, 733, and 134.The output of ADC 134 can then be compared against a range of values to determine if imaging device 100 is within acceptable design limits.","Such a test provides an ability to test the operability of each column amplifier within column amplifier 703, using a digital tester and without requiring illumination of imaging device 100.The various BIST modes for one particular embodiment are described in Table 4 below.","TABLE 4 Test Modes For Analog Blocks Negative Positive output of output of source source selector im_test selector mux 713 mux 713 im_test_en[3:0] active connected to connected to Comments 0000 0 Positive output of Negative output Normal analog mux 709 of analog mux 709 operation with CDS 0001 1 Reset level 743 Negative output Normal of analog mux operation 709 without CDS 0010 2 Reset level 743 Positive output Testing reset of analog mux 709 sampling branch of column amplifier 0011 3 atest 736 atest_n 746 Testing with external signal 0100 4 vref_n 726 vref 706 −1 V 0101 5 common 716 vref 706 −0.5 V 0110 6 common 716 common 716 0 V 0111 7 common 716 vref_n 726 0.5 V 1000 8 vref 706 vref_n 726 1 V In one particular embodiment ADC 134 is a ten-bit ADC with pipeline architecture.","The ten-bit output from ADC 134 is passed to translation unit 150.For the various dark-current calculations, four ten-bit outputs are averaged, and the most significant ten-bits of the average value are used.","Translation Unit Translation unit 150 provides color processing for imaging device 100.Parameter and mode registers associated with translation unit 150 are loaded via control bus 101.A Bayer image is received from imaging unit 130, processed by translation unit 150, and transferred to parallel interface 190 for output via parallel interface I/O 192.While processing the image, statistics are calculated by translation unit 150.After completing the processing of an image, translation unit 150 produces an interrupt to control unit 110 indicating that processing is complete and that the statistics are ready to be read by control unit 110 via control bus 101.Translation unit 150 receives the Bayer grid as input from imaging unit 130.For each received pixel, two color values are missing and need to be reconstructed.","Translation unit 150 reconstructs the RGB set for each pixel, performs various correction algorithms, and generates a YUV representation of the image.","FIG.","24 provides a block diagram 1100 illustrating translation unit 150 in greater detail.","As illustrated, translation unit 150 includes an RGB reconstruction block 1110 (for demosaicing), a CIF-to-QCIF reduction block 1130, a color and white-balance correction block 1140, a gamma look-up table (“LUT”) 1150, an RGB to YUV generation block 1160, a twenty-four bit YUV to 4:2:2 conversion block 1170, and a statistics block 1195.Statistics block 1195 includes a white balance statistics block 1150, an exposure statistics block 1185, and a G line-sum block 1190 for providing a sum of G Bayer components per line used to detect flicker.","RGB reconstruction block 1110 receives the Bayer input from imaging unit 130 and calculates all three color components for each pixel.","Median Interpolation method is selected for the Bayer-to-RGB conversion.","For green reconstruction from a pixel whose Bayer component is either red or blue, the green color component is given by the median value of the four surrounding Green Bayer components.","The median function is defined as the average between the two values that remain when a series of four values is evaluated, and the minimum and maximum values are removed.","For Blue reconstruction from a pixel whose Bayer component is either Red or Green, the Blue color component is given by the median value of the four surrounding Blue Bayer components.","For a pixel whose Bayer component is green located at a blue line, the blue value is the average of the two adjacent blue Bayer components in the same line.","Further, for a pixel whose Bayer component is green located at a red line, the blue value is the average of the two adjacent blue Bayer components in the same column.","For red reconstruction at a pixel whose Bayer component is blue, the red color component is given by the median value of the four surrounding red Bayer components.","At a pixel whose Bayer component is green located at a red line, the red value is the average of the two adjacent red Bayer components in the same line.","At a pixel whose Bayer component is green located at a blue line, the red value is the average of the two adjacent red Bayer components in the same column.","After reconstruction, the resulting RGB image is smaller than the Bayer input image by two columns horizontally, and two lines vertically.","In some embodiments, imaging device 100 provides three methods for producing a QCIF size image.","In the first method, a QCIF sized window is read from a CIF sized array.","For the method, no special processing is needed and the RGB per pixel is calculated in the same manner as for a CIF sized image.","The method provides a limited Field of View (“FOV”), but provides a good resolution.","In the second method, the array read out of pixel array 700 is down-sampled.","This method is implemented in the imaging unit 130.The third method is a Median Billinear method.","In the method, the CIF sized Bayer image is first converted to an RGB image using the Median method.","Then, if down-sampling is selected, the CIF image is reduced to a QCIF size image using a ( 1/4, 1/2, 1/4) filter—horizontally and then vertically.","In some embodiments of imaging device 100, color correction block 1140 can be used to compensate for any disparity between the RGB filters on imaging device 100 and a display connected via parallel interface I/O 192.In some instances, sensor 132 is not equally sensitive to each of the color components.","To compensate for the different sensitivity, and to emulate the human eye adjustment to different lighting conditions, the captured image undergoes a white-balance correction.","The coefficients of the color correction are combined into a single matrix, and the corrected R′G′B′ values per pixel are calculated according to the following equations: R′=CR1*R+CR2*G+CR3*B+Roffset G′=CG1*R+CG2*G+CG3*B+Goffset B′=CB1*R+CB2*G+CB3*B+Boffset In addition, Gamma Correction LUT 1150 is used to gamma correct the various pixels.","In one embodiment, Gamma correction LUT 1150 includes 512 entries, where each entry is a nine-bit value.","The data for Gamma correction LUT 1150 is loaded to RAM within imaging device 100.Gamma correction LUT 1150 receives a color component value from color correction block 1140, and uses the value as an address to access the LUT stored in RAM.","The value obtained from the LUT at the addressed location is stored until all three color components are received.","Then, the three color components are passed together to RGB-to-YUV block 1160.In YUV to 4:2:2 conversion block 1170, an input of an array of pixels, each with its own R, G and B values is reconstructed into an image of YUV 4:2:2 values.","The result is then passed to parallel interface 190.The process involves an interim conversion to a Y-Cr-Cb (4:4:4) format.","The Y-Cr-Cb (4:4:4) is calculated as follows: R′G′B′ are given at an 8-bit accuracy, in the range of 0-255, Y is limited to the range of 16-235, and Cr and Cb are limited to the range of 16-240.Then, the following equations are used to complete the conversion: Y=0.257R′+0.504G′+0.098B′+16 Cb=−0.148R′−0.291G′+0.439B′+128 Cr=0.439R′−0.368G′−0.071B′+128 The coefficients are loaded in {s,0.8} format, eight-bits of a fraction value, without integer bits, but with a single sign bit.","In the same manner, YUV can be calculated, without the clamping.","The multiply coefficients are programmable.","The offset for the luminance can be enabled or disabled.","When offset is enabled, the value of sixteen is used as offset for the luminance.","The chrominance offset is fixed at one-hundred, twenty eight.","The twenty-four bit YUV value per each pixel (Y-Cr-Cb 4:4:4), is down-sampled to a YUV 4:2:2 format.","To do this, the Y value for each pixel is retained, but each pair of consecutive luminance values (Y2n, and Y2n+1, n=0,1,2 .",".",". )","share a pair of Cb and Cr values.","The placement of the Chrominance pair is on the even numbered pixel.","FIG.","25 illustrates this down-sampling process.","The color components of the odd number pixels are zeroed out, while the color components of the even numbered pixels are calculated as follows: For first pixel in line: Cx′[0]=Cx [0] (x is either r or b) For subsequent pixels in the line: For n=1 to n, n is equal to (N/2−1), N is the line length tmp[2n]=(Cx[2n−1])+2*(Cx[2n])+(Cx[2n+1]) (x is either r or b) The result is rounded up and divided by 4: Cx′[2n]= 1/4(tmp[2n]+2) (x is either r or b) Table 14 provides a summary of calculations performed by translation unit 150 as previously discussed.","TABLE 13 Calculation Summary Var Formula Accur.","Remarks Input from C C are the Bayer grid outputs 10 bits unsigned imaging unit 130 Faulty C1 C1 = C if not faulty, 10 bits unsigned correction replacement value if faulty RGB per pel C2 C2 = (C1 + C1 + 1)/2 10 bits Average and round up C10 C10 = C2 or C1 10 bits unsigned Color C20 Color correction coefficient 8 bits + sign 8 bits value, 1 bit sign correction C21 C21 = C20 * C10 19 bits 18 bits magnitude, 1 bit sign [1, 18], C22 C22 = C21r + C21b + C21g 21 bits 21 bits 2's comp [1, 20] Toff Truncation parameter 4 bits Values 0 to 10 C23 C23 = truncated C22 21 bits Programmable C23 = {sign-ext, C22[20:t]} truncation [t − 1:0] lsbits truncated, Coff Offset for Color correction 8 bits + sign 8 bits value, 1 bit sign C24 C24 = (C23 + Coff * 2 + 1)/2 21 bits offset, round up (add 1) ignore lsbit, limit.","Lower limit 0 or 16 Loadable Upper limit (0, 511 for color correction) C25 C25 = limited C24 9 bits select bits [10:1] Gamma C30 C30 = LUT[C25] 9 bits correction RGB to Y20 Format conversion 8 bits + sign 8 bits value, 1 bit sign YUV coefficient Y21 Y21 = Y20 * C30 19 bits 18 bits magnitude, 1 bit sign [1, 18], Y22 Y22 = Y21r + Y21b + Y21g 21 bits 21 bits 2's comp [1, 20] Toff Truncation parameter 4 bits Values 0 to 10 (fixed to 8) Y23 Y23 = truncated Y22 21 bits Programmable Y23 = {sign-ext, truncation Y22[20:8]} 8 lsbits truncated Yoff Offset for conversion 8 bits + sign 8 bits value, 1 bit sign Y24 Y24 = (Y23 + Yoff * 2 + 1)/2 21 bits offset, round up (add 1) ignore lsbit, limit.","Lower limit 0 or 16 Loadable Upper limit 235/255 for Y 240/255 for U/V Y25 Y25 = limited Y24 8 bits select bits [9:1] 4:4:4 Yout Yout = Y25 8 bits to Ctmp Ctmp = (Yn − 1 + Yn * 2 + Yn + 1) 10 bits 4:2:2 Cout Cout = (Ctmp[9:1] + 1)/2 8 bits In addition, translation unit 150 collects a number of statistics during the previously discussed translations and calculations.","At the end of processing of a frame data, the resulting statistics are checked, and the different registers that control the correction algorithms are updated according to the results.","The different statistics checked by the various statistics blocks including white balance block 1180, exposure statistics block 1185, and G-line-sum block 1190.The various statistics checked can include, but are not limited to, white balance, exposure, minimum and maximum per color, and flicker.","Exposure statistics are determined by evaluating green Bayer pixels or Y output data from translation unit 150.The evaluation results in an eight-bit histogram.","There are eight seventeen-bit registers, each holding the number of pixels whose Y or Bayer value falls within the range that is defined for that bin (ten-bit Bayer components are divided by 4 before sorting).","At the end of a frame processing, programmable processing core 112 reads the histogram results and calculates the exposure, offset and gain parameters for the following frame.","Table 5 lists the eight bins into which the values are assigned to create the eight bit histogram.","TABLE 5 Exposure Histogram Bins Bin Number Range Bin #0 0-31 Bin #1 32-63 Bin #2 64-95 Bin #3 96-127 Bin #4 128-159 Bin #5 160-191 Bin #6 192-223 Bin #7 224-239 In addition, the statistics blocks of translation unit 150 detect the minimum and maximum value per each color, on the output of the Bayer-to-RGB conversion block 1110.The results can be read from registers by programmable processing core 112 at the end of the frame processing.","Further, flicker detection can be performed using the statistics blocks of translation unit 150.Flickering can be caused by a mismatch between the frequency of light impinging upon a detected image, and the frequency at which the detected image is sampled.","The effect of flickering is a modulation of the image information along the rows of pixel array 700.To eliminate flickering, the frequency at which the detected image is sample can be adjusted to coincide with the frequency of the light impinging upon the image.","In one embodiment of the present invention, an estimate of the impinging light is calculated, and the sample frequency is adjusted automatically.","In some embodiments, to obtain an estimation of the impinging frequency with high confidence, a uniform image is captured, thus avoiding detection of modulation in the image itself.","However, in other embodiments, any image is captured and flickering is automatically detected and corrected.","FIG.","26 is a flow diagram of such a flicker correction system 2205 in accordance with embodiments of the present invention.","Flicker correction system 2205 includes a feedback loop for automatically adjusting exposure to reduce flicker.","Flicker correction system 2205 includes capturing an image (block 2206) and outputting the captured image.","As is evident from the preceding discussion, the image is captured by sensor 132 and provided to translation unit 150 via ADC 134.The captured image received from ADC 134 is processed to determine if flickering is evident (block 2207).","Then, the exposure is adjusted to account for any detected flickering (block 2208).","Using the adjusted exposure parameters, the image is again captured and the adjustment process performed again.","In some embodiments, this process can be completed for one or more frames of the image.","This use of additional image information provides for more accurate detection and correction.","Further, such an iterative approach allows for continuous monitoring of flicker conditions.","In other embodiments, however, flicker detection and correction is only infrequently performed, rather than continuously.","In addition, in some embodiments, the process may adjust exposure duration in order to emphasize flicker artifacts and thereby increase the probability of correct detection and/or reduce estimation time.","Such an approach can include a manual adjustment whereby a user adjusts the capture frequency until the user detects a noticeable flicker.","After that, the flicker is automatically detected and corrected.","Alternatively, the adjustment creating the noticeable flicker can be automatically performed, and subsequently corrected.","Alternatively, a fully passive mode of operation is also possible.","In such a fully passive mode, exposure time is not altered by flicker detection mechanism until the estimation is completed.","Further, such adjustments to the exposure time can be limited to multiples of a detected flicker period.","In passive mode of operation the exposure adjustments can be introduced by other parts of the system including, for example, exposure selection sections that provide enough information for successful estimation of flicker frequency A light source, I(t), illuminating an image can have a flickering component at a frequency f that can be modeled the following way: I(t)=a·cos(2πft+φ)+b, where a, b and φ are constants.","R(n,m) is defined to be a image reflectance function that corresponds to a pixel at location (n,m) of pixel array 700.Using these definitions, the light impinging upon pixel (n,m) is given by: S ( n , m , t ) = R ( n , m ) · I ( t ) = b · R ( n , m ) + a · R ( n , m ) · cos ( 2 π ft + φ ) .","Thus, when an image is captured with exposure duration T, the value of the pixel (n,m) will be proportional to the following value: S ( n , m ) = ∫ n Δ n Δ + T S ( n , m , t ) ⅆ t = R ( n , m ) · ∫ n Δ n Δ + T I ( t ) ⅆ t , where is the time period between sampling of two adjacent rows in pixel array 700.The integration of the incident light produces: ∫ n Δ n Δ + T I ( t ) ⅆ t = b · T + a 2 π f [ sin ( 2 π f ( n Δ + T ) + φ ) - sin ( 2 π fn Δ + φ ) ] = b · T + [ a π f sin ( π fT ) ] · cos ( 2 π f Δ n + π fT + φ 2 ) , = b · T + a ′ · cos ( 2 π f ′ n + φ ′ ) , where f ′ = f · Δ a ′ ≡ a π f sin ( π fT ) ."," φ ′ ≡ π fT + φ 2 Substituting this result provides: S(n,m)=R(n,m)·b·T+R(n,m)·a′·cos(2πf′n+φ).","Thus, the resultant signal from pixel (n,m) consists of two components.","The first component, R(n,m)·b·T, does not cause flickering artifacts; whereas, the second component can change intensity across rows of pixel array 700.However, as illustrated by the preceding equation, when a′ equals zero, no flickering exists.","Therefore, since a′ is a function of exposure duration, it is possible to select such exposure duration that a′ will vanish.","Said another way, in some embodiments of the present invention, the flicker frequency is detected, and based on the detected frequency, the exposure time of pixel array 700 is adjusted to reduce flicker.","To do this, T is selected to satisfy the following constraint: f·T=k, k= .",".",".",", −2, −1, 0, 1, 2 .",".",".",".","To reduce the effect of the frequency components of the S(n,m) columns of each row are summed up to produce row averaged image S(n), where S(n) is described by the following equation: S ( n ) = ∑ m = 0 M - 1 S ( n , m ) = ( b · T + a ′ · cos ( 2 π f ′ n + φ ′ ) ) ∑ m = 0 M - 1 R ( n , m ) = R ( n ) · b · T + R ( n ) · a ′ · cos ( 2 π f ′ n + φ ′ ) .","In some embodiments, such a summation is performed by a hardware summing circuit within translation unit 150 where the green and/or any other Bayer components for each row of pixel array 700 are summed.","At the end of each row, an eighteen-bit value is sent to memory 118 of control unit 110 using a direct memory access via control bus 101.This same process is repeated for each row of pixel array 700, with each sum being stored to memory 118.Alternatively, in other embodiments, such summing can be accomplished using programmable processor core 112.In some embodiments of the present invention, all rows and pixel values therein are summed.","In other embodiments, only a subset of rows including all pixel values therein are summed, while in yet other embodiments, all rows, but only a subset of pixel therein are summed.","In yet another embodiment, a subset of the rows and a subset of the pixels therein are summed.","From these sum values, S(n) is spectrally analyzed using software executed by programmable processing core 112 to detect the presence of energy corresponding to potential flickering frequency.","FIG.","27 illustrates a graph 2211 of a typical row average.","More particularly, graph 2211 is a plot of the normalized row average 2212 verses the row number 2213 for an image detected by pixel array 700.Various spectral analysis techniques can be used on the sum information to determine a flicker frequency, if any flicker is apparent.","For purposes of illustration, operation of flicker detection system 2205 is discussed in relation to a Fourier analysis.","More particularly, two energy values, E1 and E2, are determined using the sum data.","The two values are calculated as follows: E i = F { S ( n ) } f = f i Δ F { S ( n ) } f = 0 = ∑ n = 0 N - 1 S ( n ) · ⅇ - 2 π f i Δ n ∑ n = 0 N - 1 S ( n ) = ∑ n = 0 N - 1 S ( n ) · cos ( 2 π f i Δ n ) + ∑ n = 0 N - 1 S ( n ) · sin ( 2 π f i Δ n ) ∑ n = 0 N - 1 S ( n ) .","In some embodiments, a simple comparison between the energy values E1 and E2 is used to provide an estimation of the flicker frequency.","Such estimations, however, can prove unreliable when any ordinary image is the source of the sum information rather than a controlled, constant image.","Thus, in various embodiments, a more reliable estimation of the flicker frequency is obtained by calculating a set of E1 and E2 pairs across various frames detected by pixel array 700, and comparing the variance of E1 and E2.This approach provides a number of sampling points across various images and, thus, a more robust estimation of the flicker frequency.","Improvement results because there is likely a different phase of the illumination for each frame, which provides various modulations of R(n).","After calculation of E1 and E2, corresponding to two possible flickering frequencies, the energy of the present flickering will vary depending on the phase; whereas the energy of the other frequency which depends on the R(n) only, will remain almost the same for all frames.","Therefore, the comparison between the standard deviation of E1 and E2 produces reliable results.","This procedure is presented in more detail below based on the following definition of variance: V i = 1 K ∑ k = 0 K - 1 ( E i k ) 2 - ( 1 K ∑ k = 0 K - 1 E i k ) 2 , where K is the number of frames that were captured and Eik is the energy value corresponding to frequency fi of the k-th image.","To improve the performance, frames can be captured at two or more different exposures that each exhibit flickering.","As an example, the following equation describes exposure times creating such a dual flicker: T = ( l + 1 2 ) · 1 f i , i = 1 , 2 ; l = 0 , 1 , 2 , … When both V1 and V2 are below some predefined threshold, no flickering has been detected.","Otherwise, the estimated flickering frequency is the frequency that produced larger Vi.","FIG.","28 illustrates a flow diagram 2241 of a method for detecting and correcting flicker in accordance with the present invention.","Following flow diagram 2241, a frame is captured from pixel array 700 (block 2243).","The lines of the frame are summed, and E1 and E2 are calculated as previously described (block 2247).","Next, it is determined if a sufficient amount of information has been gathered to make a reliable determination of flicker frequency (block 2249).","In one embodiment, satisfactory results were obtained when seventeen frames of data are obtained before determining the flicker frequency.","If not, more information is gathered by processing additional frames.","Alternatively, where sufficient information has been gathered, V1 and V2 are calculated using the gathered information (block 2251).","The determined variance value is set equal to the maximum of V1 and V2, and the frequency is set to the frequency fi at which the maximum variance was detected (block 2253).","Where the determined variance is less than a predefined threshold level D (block 2257), it is determined that no flicker is detected (block 2259), and the process is repeated to detect a change in conditions that results in flicker.","Alternatively, where the determined variance is greater than the a predefined threshold level D (block 2257), a flicker frequency of fi is detected (block 2261) and the exposure duration is constrained to a multiple of 1/fi (block 2263) to correct the detected flicker.","In some embodiments, the previously described approach is simplified to reduce computational complexity.","More particularly, instead of calculating the exact Fourier component, it is replaced by an approximation of the energy value according to the following equation: E ^ i = ∑ n = 0 N - 1 S ( n ) · SC ( 2 π f i Δ n ) + ∑ n = 0 N - 1 S ( n ) · SS ( 2 π f i Δ n ) ∑ n = 0 N - 1 S ( n ) , where SC ( x ) = sign ( cos ( x ) ) = { 1 , - π 2 ≤ x ≤ π 2 - 1 , π 2 ≤ x ≤ 3 π 2 and SS ( x ) = sign ( sin ( x ) ) = { 1 , 0 ≤ x ≤ π - 1 , π ≤ x ≤ 2 π .","However, such a computational reduction can cause degradation in frequency estimation performance.","Nevertheless, even after such degradation, satisfactory results of flicker detection and correction can be obtained.","Moreover, in some embodiments, only one of the components is used during the estimation process.","Such an approach results in insignificant performance degradation which may be overcome by increasing number of considered frames K. In yet other embodiments where greater confidence in the estimated flickering frequency is desired, a more complex approach is utilized.","In such approaches, various measures of confidence can be implemented to prevent frequency switches (block 2263) when an incorrect estimate of flicker frequency is detected.","Further, in some embodiments, residual flickering (i.e.","flickering due to slight variance around the flickering frequency) can be eliminated.","More particularly, one such improvement increases the probability of a correct estimation by utilizing more pixel values in the various calculations.","Instead of using just a standard deviation of the energy that corresponds to possible flickering frequency to make decision, one may base the estimation on a number, L, of consequent standard deviations.","The following equations illustrate such an approach: W 1 = ∑ l = 0 L - 1 V 1 l V 2 l W 2 = ∑ l = 0 L - 1 V 2 l V 1 l C 1 = ∑ l = 0 L - 1 I ( V 1 l > V 2 l ) C 2 = ∑ l = 0 L - 1 I ( V 2 l > V 1 l ) M = max l { ∑ k = 0 K - 1 E 1 k , 1 + E 2 k , l } where I(·) is an indicator function defined below: I ( Condition ) = { 1 , if Condition is true 0 , otherwise , and Vxl,Exk,l denotes the l-th standard deviation and the energy of k-th frame in l-th run, respectively.","Note that the total number of frames that should be acquired prior the decision is K·L.","The decision is made based on the following set of rules: 1.If M is below a predefined flickerless threshold D0 no flicker artifact was detected.","2.If W1>D1∩C1>D2, flicker at f1 was detected.","3.If W2>D1∩C2>D2, flicker at f2 was detected.","4.Otherwise, the information is ambiguous and no decision is taken.","This method reduces drastically the probability of both misdetection and incorrect detection.","However, the delay and probability of false alarm are increased, which can be less critical in some applications.","Further, to prevent the situation when occasional wrong frequency estimation changes the exposure to a value that leads to flicker artifact, some embodiments include additional mechanisms to reduce this possibility.","One such approach for avoiding this situation includes an additional counter, referred to as a confidence counter.","The confidence counter is initialized to zero.","Whenever flicker at a first frequency, f1, is estimated the confidence counter is increased by one for each estimation until a limit, B, is reached.","If estimated flicker frequency is a second frequency, f2, the confidence counter is decreased by one for each estimation until a limit, negative B, is reached.","In a case of ambiguity, no change is made to the confidence counter.","When the confidence counter is positive, the global estimation is f1 and exposure is limited to multiple of 1/f1.When the confidence counter is negative, the global estimation is f2 and exposure is limited to multiple of 1/f2.If the confidence counter is zero previous estimation is used.","This reduces the possibility of spurious data creating an unstable exposure time.","Parallel Interface Parallel interface 190 produces the video output of imaging device 100.parallel interface 190 includes a ten-bit parallel data bus, a vertical frame signal, a clock, and a qualifying signal.","Parallel interface 190 can also produce a serial output selectable by programming a register.","When operating in a parallel mode, imaging device 100 outputs a video image in a parallel format.","The parallel format includes a clock signal along with data signals and qualifying signals each synchronized to the clock signal.","In some embodiments of imaging device 100, the effective edge of the clock signal (i.e., rising or falling edge) is programmable.","The data signals include a ten-bit wide data bus carrying either the Bayer grid data, or the YUV 4:2:2 output data, where the type of data is determined by monitoring a programmed register.","The eight-bit YUV data is presented on the most significant data signals (i.e., bits three through ten of the ten-bit bus).","On each effective edge of the clock, a single data byte is transferred on the data bus where a horizontal qualifying signal is asserted.","The horizontal qualifying signal is asserted only during valid image data, or to qualify optional start-of-line and end-of-line markers.","A vertical qualifying signal is asserted on the effective edge of the clock to mark the beginning of the first line in a new image frame.","The duration of the vertical qualifying signal is programmable via a register implemented as part of imaging unit 130.FIG.","29 illustrates the vertical qualifying signal, VALIDV, and the horizontal qualifying signal, VALIDH, in relation to a pixel array where the active image portion of the array is designated.","As illustrated, VALIDV is a vertical synchronization signal whose effective edge (programmable by setting a register value) marks the beginning of the first line in a new image frame.","FIG.","30 illustrates the signals in relation to data signals referred to a valid data, and FIGS.","34 and 35 illustrate a bitwise view of the signals in relation to Bayer and YUV data, respectively.","As illustrated, when sending out the data in Bayer-grid format, a “RED” line will always be the first line in the frame.","When sending out the data in YUV output format, the order of the color components is programmable via a register value.","Control unit 110 controls the frame rate output via parallel interface 190 via a C_outframe signal.","Via the signal, control unit 110 enables or disables the output of a full frame.","When the signal is deasserted, VALIDH and VALIDV signals are stuck at an inactive state and the data signals hold the background value throughout the disabled frame time.","By deasserting C_outframe, a frame of data can be skipped as illustrated in FIG.","33.The type of data output via parallel interface 190 is programmable.","When Bayer data is output, the data volume is an 8/9/10 bit word.","If data markers are added, eight more bytes per line must be taken into consideration in the image buffer that is allocated for the application.","There are two types of BAYER data lines: red lines and blue lines.","Red lines carry green and red Bayer components.","The green and red data bytes are interleaved where the even numbered bytes (0, 2 to 2n) are green, and the odd numbered bytes (1, 3 to 2n+1) are red.","On the other hand, blue lines carry blue and green Bayer components, where the components are interleaved with the even numbered bytes (0, 2 to 2n) being blue, and the odd numbered bytes (1, 3 to 2n+1) being green.","When sending Bayer data out on parallel interface 190, a red line is the first line in the frame.","When YCrCb 4:2:2 data is output, the data volume is two bytes per pixel and no extra pixels are transmitted beyond the CIF/QCIF size.","If data markers are added, eight more bytes per line must be taken into consideration in the image buffer that is allocated for the application.","The data in the YCrCb 4:2:2 output format is ordered in several configurations, according to the setting in a programmable color register.","The following illustrates four possible color orders: 00—SOL, Y0, U0, Y1, V0, Y2, U2, Y3, V3, to Yn-2, Un-2, Yn-1, Vn-2, EOL 01—SOL, Y0, V0, Y1, U0, Y2, V2, Y3, U3, to Yn-2, Vn-2, Yn-1, Un-2, EOL 10—SOL, U0, Y0, V0, Y1, U2, Y2, V3, Y3, to Un-2, Yn-2, Vn-2, Yn-1, EOL 11—SOL, V0, Y0, U0, Y1, V2, Y2, U3, Y3, to Vn-2 Yn-2, Un-2, Yn-1, EOL In some embodiments, extra data markers are inserted at the beginning and end of each image line.","This insertion is optional and can be selected via a programmable register.","The following are four examples of data marker types: Start of First Line marker—This marker precedes the data of the first line in a frame.","Start of Line Marker—This marker precedes the data of each line in a frame, except for the first line.","When Bayer output mode is selected, bit #5 of the marker content differentiates between a “red” line and a “blue” line (0 and 1 respectively).","End of Last Line Marker—This marker immediately follows the data of the last line in a frame.","End of Line Marker—This marker immediately follows the data of each line in a frame, except for the last line.","In addition to outputting data in a parallel mode, parallel interface 190 can format the data for serial output.","The Transmitted data size for serial output mode is eight bits per transmitted “word”.","Bayer output provides eight bits per pixel, and YUV output is sixteen bits per pixel.","The bytes are transmitted with the most significant bits first.","During a continuous transfer, the most significant bit of byte N immediately follows the least significant bit of byte N−1.In some embodiments, imaging device 100 is integrated into systems where the Host is pin limited.","For example, in one particular embodiment, the host is a Digital Signal Processor capable of receiving video via a two or three pin interface.","In such systems, imaging device 100 can be configured to provide video output via a serial interface.","In some embodiments the serial mode provided by imaging device 100 relies on elements consistent with ITU-RBT.656-4 standard.","ITU-RBT.656-4 standard recommendation is available from the International Telecommunication Union, the entirety of which is incorporated herein by reference for all purposes.","In one particular embodiment, imaging device 100 can be configured to operate in different serial output modes: a clock-qualify mode and a synchronous serial interface (“SSI”) mode.","Such functionality provides an ability to function in low cost, reduced pin-count environments.","In contrast to ITU-RBT.656-4 standard, in embodiments of the present invention where data markers are used, the EAV and SAV markers are different.","In particular, the second most significant bit (i.e.","bit six) of the markers is a special frame marker used to indicate the start of the first line of a frame and the end of the last line of a frame.","Further, bit five indicates the Bayer color, where a logical one indicates blue and a logical 0 indicates red.","In the serial output modes, either two or three pins of the thirteen-bit parallel video interface 192 are utilized to output the serial video data.","Further, in each of the serial output modes, the frequency of the clock, DSCLK 2920, used to output the serial data is eight times (“8×”) that of a clock, CLK, used to output parallel video data via parallel video interface 192.In some embodiments, DSCLK 2920 is provided via an external pin to imaging device 100.In such cases, the input clock source is internally divided to provide clocking useful for operating parallel video interface 192 when both parallel and serial video data are being provided from imaging device 100.In other embodiments, an internal phase lock loop is used to generate the various output clocks for use in relation to both the serial and parallel output modes of imaging device 100.In operation, parallel video data is received from translation unit 150.Where a serial output mode is selected via a programmable register, the parallel data is shifted out via the designated serial pins of parallel video interface 192 using the 8× clock.","This shifting occurs by transmitting the most significant bit of any parallel data first and following in succession with bits of decreasing significance.","Clock-qualify mode is a two wire interface with clock and output data signals, DSCLK 2920 and DSDATA 2940.Imaging device 100 acts as a master of the interface.","As such, imaging device 100 activates DSCLK 2920 when there is valid data to be sent out via DSDATA 2940.FIG.","34 illustrates the clock-qualify mode where data can be output at two rates represented by DSCLK/2 2950 and DSCLK 2920, both being synchronized to CLK of imaging device 100.As illustrated, DSDATA 2940 is provided by imaging device 100 on the rising edge of DSCLK/2 2950 and latched by the host on the falling edge of DSCLK/2 2950.It should be recognized that twice the transfer rate is possible where the same approach is used in relation to DSCLK 2920.Using the approach, eight bits are transferred from the most significant bit to the least significant bit.","In some embodiments, one deasserted cycle of DSCLK 2920 or DSCLK/2 2950 indicates a word boundary, while in other embodiments, word boundaries are contiguous allowing for a continuous transfer of data based on a continuing period of DSCLK 2920 or DSCLK/2 2950.SSI mode is a three wire interface, with clock (DSCLK 2920), output data (DSDATA 2940) and frame signals (DSFRAME 2930).","Imaging device 100 acts as a master of the interface, producing all three signals.","FIG.","35 illustrates the operation of SSI mode with DSCLK 2920, DSFRAME 2930, and DSDATA 2940.As illustrated, DSFRAME 2930 is asserted by imaging device 100 prior to a rising edge of DSCLK 2920.The host accepts the asserted DSFRAME 2930 on a rising edge 2910 of DSCLK 2920.On the eight following rising edges of DSCLK 2920, one of the eight bits of a byte transfer are provided by imaging device 100 as DSDATA 2940 to a receiving host.","The bits are provided with the most significant bit of the by first followed sequentially by bits of decreasing significance.","Testability Unit Imaging device 100 can additionally include various mechanisms for performing tests of the device.","Using one such mechanism, a built-in-self-test (BIST) is implemented to facilitate production testing of ADC 134 and the various DAC circuitry.","The BIST circuit creates a sequence of values that are passed to ADC 134 after application of the gain and two offset parameters as previously discussed.","The output from sensor 132.With this canned value presented to ADC 134, the outputs of ADC 134 is checked against a known output.","If the output falls within a programmable tolerance level, imaging device 100 is acceptable.","If not, imaging device 100 is rejected.","In addition, a BIST mechanism for sensor 132 is implemented.","A minimum and maximum value is defined for each color within sensor 132.Sensor 132 is illuminated by a monochrome color, and the resulting image is tested.","The value of each pixel is checked.","At the end of the frame, three values can be read per each color.","The number of pixels below the minimum value, the number of pixels above the maximum value, and the number of pixels falling within the minimum and maximum values is determined.","Additionally, BIST mechanisms are implemented to test various RAMs and registers within imaging device 100 and to return pass or fail results.","The invention has now been described in detail for purposes of clarity and understanding.","However, it will be appreciated that certain changes and modifications may be practiced within the scope of the appended claims.","Accordingly, it should be recognized that many other systems, functions, methods, and combinations thereof are possible in accordance with the present invention.","Thus, although the invention is described with reference to specific embodiments and figures thereof, the embodiments and figures are merely illustrative, and not limiting of the invention.","Rather, the scope of the invention is to be determined solely by the appended claims."]],"string":"[\n [\n \"Patent application cmos imager for cellular applications and methods of using such\",\n \"Systems, methods and devices related to detecting and transmitting images.\",\n \"Imaging systems and devices, as well as methods of using such that are provided herein include flicker detection and/or correction; and/or built-in self test associated with various analog circuitry in the imaging devices; and/or power reduction ability; and/or pixels with charge evacuation functionality; and/or a parallel to serial conversion unit and associated serial output interface; and/or other advanced functionality.\"\n ],\n [\n \"1.A CMOS imaging device, the CMOS imaging device comprising: a pixel array; a selector, wherein the selector is operable to select between an input derived from the pixel array and at least one reference input, and wherein the selection between the input derived from the pixel array and the at least one reference input is at least in part based on a signal derived from a digital domain; and an analog to digital converter, wherein the analog to digital converter is operable to convert a signal derived from the selector to the digital domain.\",\n \"2.The CMOS imaging device of claim 1, wherein the signal derived from the selector is an output of the selector that is passed through an analog processing circuit.\",\n \"3.The CMOS imaging device of claim 2, wherein the analog processing circuit consists of one or more circuits selected from a group consisting of: a level shift circuit, a programmable offset circuit, and a programmable gain amplifier.\",\n \"4.The CMOS imaging device of claim 2, the CMOS imaging device further comprising: an analog processing selector, wherein the analog processing selector is operable to bypass one or more portions of the analog processing circuit.\",\n \"5.The CMOS imaging device of claim 1, the CMOS imaging device further comprising: a comparator receiving an output from the analog to digital converter, wherein the comparator is selected from a group consisting of: a programmable hardware comparator, a programmable software comparator, and a hardwired comparator.\",\n \"6.The CMOS imaging device of claim 1, wherein the pixel array includes at least one pseudo-pixel, the CMOS imaging device further comprising: a pixel array selector, wherein the pixel array selector is operable to select the at least one pseudo-pixel, and wherein the input derived from the pixel array is derived from the pseudo-pixel.\",\n \"7.The CMOS imaging device of claim 6, the CMOS imaging device further comprising: a column amplifier, wherein the column amplifier receives an output from the pseudo-pixel, and amplifies the output from the pseudo-pixel; and wherein the input derived from the pixel array is derived via the column amplifier.\",\n \"8.The CMOS imaging device of claim 1, wherein the reference input to the selector is a pixel under reset, the CMOS imaging device further comprising: a circuit for selecting the pixel under reset; and wherein the selector is operable to provide a differential pair consisting of the input derived from the pixel array and the pixel under reset.\",\n \"9.The CMOS imaging device of claim 1, wherein at least one of the reference inputs to the selector is tied to a pad of the CMOS imaging device.\",\n \"10.The CMOS imaging device of claim 1, wherein at least one of the reference inputs to the selector comprises one or more voltages generated internal to the CMOS imaging device.\",\n \"11.The CMOS imaging device of claim 1, wherein the input derived from the pixel array and the at least one reference input are both differential inputs.\",\n \"12.A method for testing analog functionality of a CMOS imaging device in the digital domain, the method comprising: providing a pixel array; providing a selector, wherein the selector is operable to select between an input derived from the pixel array and at least one reference input; providing an analog to digital converter, wherein the analog to digital converter is operable to convert a signal derived from the selector to the digital domain; asserting a selection signal in the digital domain, wherein a derivative of the selection signal is input to the selector and at least in part controls the selection between the input derived from the pixel array and the reference input; and analyzing an output of the analog to digital converter to verify that the CMOS imaging device is functional.\",\n \"13.The method of claim 12, wherein the reference input is a differential input, and wherein the reference input is a combination of two signals selected from a group consisting of: a common voltage, a negative reference voltage, and a positive reference voltage.\",\n \"14.The method of claim 13, wherein asserting the selection signal causes the reference input to be selected, the method further comprising: providing an analog processing circuit, wherein the analog processing circuit receives the reference input from the selector, and processes the reference input, and wherein the signal derived from the selector is the processed reference input.\",\n \"15.The method of claim 14, wherein the analog processing circuit comprises a first circuit and a second circuit, the method further comprising: bypassing the first circuit.\",\n \"16.The method of claim 12, wherein the analyzing the output of the analog to digital converter comprises comparing the output of the analog to digital converter to determine if the output is within a predefined range.\",\n \"17.The method of claim 12, wherein the pixel array includes at least one pseudo-pixel, the method further comprising: selecting the pseudo-pixel, wherein an output of the pseudo pixel is not sensitive to an illumination of the CMOS imaging device, and wherein the input derived from the pixel array is derived from the output of the pseudo-pixel; and wherein asserting the selection signal causes the input from the pixel array to be selected.\",\n \"18.The method of claim 17, the method further comprising: providing a column amplifier, wherein the input derived from the pixel array is derived via the column amplifier.\",\n \"19.The method of claim 12, wherein the reference input is a differential input, and wherein at least one differential signal of the reference input is generated external to the CMOS imaging device.\",\n \"20.The method of claim 12, wherein the input derived from the pixel array is a differential input, wherein the at least one reference input comprises a reset level, and wherein the selection between the input derived from the pixel array and the reference input includes selecting one of the differential signals associated with the input derived from the pixel array and the reset level, and providing the input derived from the pixel array and the reset level as a differential pair.\",\n \"21.The method of claim 20, the method further comprising: resetting at least one pixel in the pixel array; and selecting the at least one reset pixel, wherein the input derived from the pixel array is derived from the at least one reset pixel.\",\n \"22.A CMOS imager, wherein the CMOS imager comprises: an image sensor, wherein the image sensor includes one or more rows of pixels, and wherein each of the pixels provides a pixel value; a storage element; a summing circuit, wherein the summing circuit is operable to sum the pixel values for each of the one or more rows to create row sums, and to store the row sums to the storage element; and a programmable core, wherein the programmable core includes instructions executable by the programmable core to: determine a first energy value and a second energy value associated with the row sums; compare the first energy value and the second energy value to estimate a flicker frequency; and constrain an exposure duration associated with the image sensor to approximate a multiple of the flicker duration.\",\n \"23.The CMOS imager of claim 22, wherein the summing circuit is further operable to calculate row sums for a plurality of frames, wherein each of the plurality of frames includes pixel values from the two or more rows of pixels, and wherein the instructions are further executable by the programmable core to: determine a first energy value and a second energy value associated with the row sums for each frame; calculate a variance between first energy values across the plurality of frames; calculate a variance between second energy values across the plurality of frames; and wherein the comparing the first energy value and the second energy value to estimate a flicker frequency comprises comparing the variance of the first energy values with the variance of the second energy values.\",\n \"24.The CMOS imager of claim 23, wherein the instructions are further executable by the programmable core to: compare the variance of the first energy values with the variance of the second energy values, wherein the larger is a greater variance; and comparing the greater variance with a predefined threshold, wherein the constraining the exposure frequency is done when the greater variance is greater than the predefined threshold.\",\n \"25.The CMOS imager of claim 24, wherein the CMOS imager further comprises: a confidence counter, wherein the confidence counter is incremented whenever the estimated flicker frequency is approximately a first frequency, and decremented whenever the estimated flicker frequency is not approximately the first frequency.\",\n \"26.The CMOS imager of claim 25, wherein the constraining the exposure frequency is done when the confidence counter changes its sign.\",\n \"27.The CMOS imager of claim 22, wherein the CMOS imager further comprises: an interrupt associated with the summing circuit, wherein assertion of the interrupt indicates that summing for the one or more rows of pixels is complete.\",\n \"28.The CMOS imager of claim 27, wherein the instructions are further executable by the programmable core to receive the interrupt.\",\n \"29.The CMOS imager of claim 22, wherein the summing circuit is further operable to calculate row sums for a plurality of frames, wherein each of the plurality of frames includes pixel values from the two or more rows of pixels, and wherein the CMOS imager further comprises: an interrupt associated with the summing circuit, wherein assertion of the interrupt indicates that summing for the two or more rows of pixels for a frame is complete; and wherein the instructions are further executable by the programmable core to: receive the interrupt; determine a first energy value and a second energy value associated with the row sums for each frame; determine if a sufficient number of frames have been received to calculate a variance between first energy values and a variance between second energy values; calculate the variance between first energy values across the plurality of frames; calculate the variance between second energy values across the plurality of frames; and wherein the comparing the first energy value and the second energy value to estimate a flicker frequency comprises comparing the variance of the first energy values with the variance of the second energy values.\",\n \"30.The CMOS imager of claim 22, wherein determining the first energy value and the second energy value comprises calculating an exact Fourier component for the first energy value and for the second energy value.\",\n \"31.A method for automatically detecting flicker in an imaging device, the method comprising: providing an imaging device, wherein the imaging device includes an image sensor with two or more rows of pixels, and wherein each of the pixels provides a pixel value; retrieving at least a portion of the two or more rows of pixels, and summing the pixel values to create a row sum for each of the rows of pixels; determining a first energy value and a second energy value associated with the row sums; and comparing the first energy value and the second energy value to estimate a flicker frequency.\",\n \"32.The method of claim 31, the method farther comprising: constraining an exposure duration associated with the image sensor to approximate a multiple of the flicker frequency period.\",\n \"33.The method of claim 31, wherein determining the first energy value and the second energy value comprises calculating an approximate Fourier component for the first energy value and for the second energy value.\",\n \"34.The method of claim 31, wherein the retrieving at least a portion of each of the two or more rows of pixels, sunning the pixel values, and determining a first energy value and a second energy value is performed for each of a plurality of frames, the method further comprising: calculating a variance between first energy values across the plurality of frames; calculating a variance between second energy values across the plurality of frames; and wherein the comparing the first energy value and the second energy value to estimate a flicker frequency comprises comparing the variance of the first energy values with the variance of the second energy values.\",\n \"35.The method of claim 34, the method further comprising: comparing the variance of the first energy values with the variance of the second energy values, wherein the larger is a greater variance; and comparing the greater variance with a predefined threshold, wherein the constraining the exposure duration is done when the greater variance is greater than the predefined threshold.\",\n \"36.The method of claim 35, the method further comprising: incrementing a confidence count whenever the estimated flicker frequency is approximately a first frequency, and decrementing the confidence count whenever the estimated flicker frequency is not approximately the first frequency.\",\n \"37.The method of claim 36, wherein the constraining the exposure duration is done when the confidence counter changes sign.\",\n \"38.The method of claim 31, wherein the summing the pixel values to create a row sum for each of the rows of pixels is done by a summing circuit included in the imaging device, the method further comprising: issuing an interrupt associated with the summing circuit, wherein the interrupt indicates that summing for the two or more rows of pixels is complete.\",\n \"39.The method of claim 38, the method further comprising: receiving the interrupt, wherein determining the first and the second energy values is at least in part triggered by the interrupt.\",\n \"40.An imaging device, wherein the imaging device comprises: a pixel array, wherein the pixel array comprises a plurality of rows, wherein each of the plurality of rows comprises a plurality of pixels, and wherein each of the plurality of pixels provides a pixel value; a summing circuit, wherein the summing circuit is operable to sum the pixel values for one or more of the plurality of rows to create row sums; an interrupt associated with the summing circuit, wherein the interrupt indicates that summing is complete for one or more of the plurality of rows; and a programmable core, wherein the programmable core includes instructions executable by the programmable core to: receive the interrupt; determine a first energy value and a second energy value associated with the row sums for each frame of a plurality of frames; calculate a variance between first energy values across the plurality of frames; calculate a variance between second energy values across the plurality of frames; and compare the variance of the first energy values with the variance of the second energy values, wherein the larger is a greater variance.\",\n \"41.The imaging device of claim 40, wherein the instructions are further executable by the programmable core to: constrain the exposure duration to a multiple of the flicker frequency period.\",\n \"42.A CMOS imager, wherein the CMOS imager comprises: an image sensor; a sub-sample control system, wherein the sub-sample control system is operable to identify one or more signals from the image sensor that are not processed; an analog processing circuit, wherein the analog processing circuit receives the one or more signals from the image sensor at one or more inputs; and a processing control system, wherein the processing control system is operable to limit switching of the one or more inputs.\",\n \"43.The CMOS imager of claim 42, wherein the analog processing circuit comprises a column amplifier, and wherein the one or more inputs are inputs to the column amplifier.\",\n \"44.The CMOS imager of claim 42, wherein the analog processing circuit comprises a level shift circuit, and wherein the one or more inputs are inputs to the level shift circuit.\",\n \"45.The CMOS imager of claim 42, wherein the analog processing circuit comprises a programmable offset circuit, and wherein the one or more inputs are inputs to the programmable offset circuit.\",\n \"46.The CMOS imager of claim 42, wherein the processing control system is operable to maintain a common mode feedback loop of an amplifier associated with the analog processing circuit in an active state, while limiting switching of the one or more inputs.\",\n \"47.The CMOS imager of claim 42, the CMOS imager further comprising: a digital processing circuit, wherein the processing control system is further operable to disable the digital processing circuit.\",\n \"48.A method for processing images in a power sensitive application, the method comprising: providing a CMOS imager, wherein the CMOS imager includes a pixel array, and an analog processing circuit; defining a first portion and a second portion of the pixel array, wherein the first portion of the pixel array includes a output image area and the second portion of the pixel array includes a dropped image area; receiving a pixel signal from the second portion of the pixel array; and placing the analog processing circuit in a standby mode, wherein the pixel signal from the second portion of the pixel array is not processed.\",\n \"49.The method of claim 48, wherein placing the analog processing circuit in the standby mode comprises: limiting switching of one or more inputs to the analog processing circuit.\",\n \"50.The method of claim 48, wherein placing the analog processing circuit in the standby mode comprises: electrically coupling one or more inputs of the analog processing circuit to a static voltage, wherein switching of the analog processing circuit is reduced.\",\n \"51.The method of claim 48, wherein placing the analog processing circuit in the standby mode comprises: electrically coupling one or more inputs of the analog processing circuit to a static voltage, while maintaining switching associated with a common mode feedback loop of one or more amplifiers associated with the analog processing circuit, wherein switching of the analog processing circuit is reduced, and bias currents are reduced while maintaining the one or more amplifiers at its inertial state.\",\n \"52.The method of claim 48, the method further comprising: receiving a pixel signal from the first portion of the pixel array; and placing the analog processing circuit in an operational mode, wherein the pixel signal from the first portion of the pixel array is processed.\",\n \"53.The method of claim 48, wherein the power sensitive application comprises video transmission via a cellular telephone.\",\n \"54.The method of claim 48, wherein the power sensitive application comprises image detection and processing using a mobile device.\",\n \"55.The method of claim 54, wherein the mobile device is selected from a group consisting of a cellular telephone, a personal digital assistant, and a laptop computer.\",\n \"56.The method of claim 48, wherein the CMOS imager further comprises a digital processing circuit, the method further comprising: disabling the digital processing circuit, wherein the pixel signal from the second portion of the pixel array is not processed.\",\n \"57.The method of claim 56, wherein disabling the digital processing circuit comprises: gating at least one clock driving the digital processing circuit.\",\n \"58.The method of claim 52, wherein the CMOS imager further comprises a digital processing circuit, the method further comprising: enabling the digital processing circuit, wherein the pixel signal from the first portion of the pixel array is processed.\",\n \"59.The method of claim 48, wherein the analog processing circuit comprises: a column amplifier; and a level shift circuit.\",\n \"60.The method of claim 59, wherein the analog processing circuit further comprises: a black level offset; a programmable gain amplifier; and an ADC.\",\n \"61.The method of claim 56, wherein the digital processing circuit includes a translation unit.\",\n \"62.A method for reducing power consumption in an imaging device, the method comprising: providing an imaging device including an image sensor and an analog processing circuit; sub-sampling the image sensor, wherein a first portion of the image sensor is not processed; receiving an image signal from the first portion of the image sensor; and placing the analog processing circuit in a standby state, wherein the image signal from the first portion is not processed.\",\n \"63.The method of claim 62, wherein placing the analog processing circuit in a standby state comprises: limiting switching of one or more inputs to the analog processing circuit.\",\n \"64.The method of claim 62, wherein the analog processing circuit comprises: a level shift circuit; a programmable offset circuit; and a programmable gain amplifier.\",\n \"65.The method of claim 64, wherein placing the analog processing circuit in a standby state comprises: electrically coupling one or more inputs of the analog processing circuit to a static voltage, wherein switching of the analog processing circuit is reduced.\",\n \"66.The method of claim 65, wherein placing the analog processing circuit in a standby state further comprises: maintaining switching associated with a common mode feedback loop of one or more amplifiers associated with the analog processing circuit, wherein the one or more amplifiers are maintained in an inertial state, and a bias current associated with the amplifiers is reduced.\",\n \"67.The method of claim 62, wherein sub-sampling the image sensor is done to create a bandwidth matched image output from the imaging device.\",\n \"68.The method of claim 62, the method further comprising: receiving an image signal from a second portion of the image sensor, wherein the second portion the image sensor is processed; and placing the analog processing circuit in an operational mode, wherein the image signal from the second portion is processed.\",\n \"69.The method of claim 68, wherein placing the analog processing circuit in an operational mode comprises electrically coupling one or more inputs of the analog processing circuit to the image signal from the second portion.\",\n \"70.The method of claim 62, wherein the imaging device further comprises a digital processing circuit, the method further comprising: disabling the digital processing circuit.\",\n \"71.A CMOS imager, wherein the CMOS imager comprises an image sensor; a parallel to serial data conversion unit, wherein the parallel to serial data conversion unit receives an image derived from the image sensor and converts the image to a serial data stream; and a serial output interface, wherein the serial output interface provides access to the serial data stream.\",\n \"72.The CMOS imager of claim 71, wherein the image sensor comprises one or more pixels, and wherein the one or more pixels include a charge evacuation element.\",\n \"73.The CMOS imager of claim 71, wherein the serial output interface includes a clock signal and a data signal, and wherein the clock signal is only active when valid data is presented on the data signal.\",\n \"74.The CMOS imager of claim 71, wherein the serial output includes a clock signal, a data signal, and a qualifying signal, and wherein the qualifying signal indicates the presence of valid data on the data signal.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>The present invention relates generally to systems and methods for detecting and/or transmitting images.\",\n \"More particularly, the present invention relates to detecting and transmitting images in relation to a cellular telephone.\",\n \"Image processors typically detect light reflected off of a subject, and convert the detected light to an electrical image.\",\n \"The electrical image is then sampled at a frequency that is the reciprocal of the integration time.\",\n \"In many cases, such a frequency is different from the frequency of light reflected off the subject.\",\n \"In such cases, a modulation of the displayed image can result.\",\n \"Such modulation appears as a flicker and can be very distracting.\",\n \"To overcome this problem, some image processors allow for manual selection between different integration times that allow a user to reduce the flicker.\",\n \"For example, where an image processor will be used in a 60 Hz and a 50 Hz lighting environment, a manual selection between an integration time associated with either 60 Hz or 50 Hz can be provided.\",\n \"This alleviates some flicker, but requires adjustment from a user.\",\n \"Furthermore, such an approach is limited to reducing flicker in a limited number of pre-determined environments, and thus may not be able to address various flicker situations.\",\n \"This is particularly problematic for mobile devices that are used in ever changing environments.\",\n \"Further, a limited reservoir of power typically exists for mobile devices.\",\n \"As such, power consuming imaging applications are often incompatible with mobile devices.\",\n \"Various approaches exist to increase the amount of power available to mobile devices, such that the power requirements of an imaging application are not prohibitive.\",\n \"However, such approaches often are both expensive and can result in increased dimensions of a mobile device.\",\n \"In some cases, light detected by an image processor is detected by a pixel array and a variety of analog processing circuitry.\",\n \"Output from the pixel array representing the amount of detected light is processed by analog circuitry to produce an image which is subsequently converted from the analog domain to the digital domain.\",\n \"The digital image is subsequently provided to a monitor of some sort for viewing.\",\n \"Testing such an imaging device can include reflecting light off a known image, and determining if the image was properly acquired and/or processed by the imaging device.\",\n \"This method provides an effective approach for testing such devices, however, the method is subject to a number of variables including lighting and image stability which can effect testing of the imaging device.\",\n \"Providing such a test, and controlling for such variables can be costly and time consuming.\",\n \"Further, such an approach tests the imaging device holistically, and is limited in its ability to identify sub-components of the imaging device which may have failed.\",\n \"Hence, for at least the aforementioned reasons, there exists a need in the art to provide advanced systems, methods and devices for detecting and/or transmitting images.\"\n ],\n [\n \"<SOH> BRIEF SUMMARY OF THE INVENTION <EOH>The present invention provides systems, devices, and methods for imaging.\",\n \"More particularly, the present invention provides devices and methods for capturing images, formatting the images, and/or transmitting the images via a cellular telephone network or other wireless network.\",\n \"Various aspects of the present invention include an imaging device with one or more of the following units: a control unit, a serial interface unit, a parallel interface unit, an imaging unit, and a translation unit.\",\n \"In some embodiments, flicker detection and/or correction functionality is incorporated.\",\n \"Additionally, in some embodiments, a built-in self test associated with various analog circuitry in the imaging device is included.\",\n \"Further, in some embodiments, systems and methods for reducing power consumption by various analog circuitry within the imaging device are included.\",\n \"Yet further, in some embodiments, a parallel to serial conversion unit and associated serial output interface are included.\",\n \"These and many other novel aspects of the present invention are provided in greater detail in the proceeding portions of this document.\",\n \"One particular embodiment of the present invention provides an imaging device and methods for using such that provide for advanced testing ability.\",\n \"In one aspect, an imaging device includes a pixel array.\",\n \"The imaging device further includes a selector that is operable to select between an input derived from the pixel array and at least one reference input.\",\n \"Such selection between the input derived from the pixel array and the at least one reference input is based at least in part on a signal derived from a digital domain.\",\n \"Further, the imaging device includes an analog to digital converter that is operable to convert a signal derived from the selector to the digital domain.\",\n \"In some cases, such an embodiment provides an ability to verify the functionality of analog circuitry within the imaging device using a digital tester.\",\n \"In some aspects of the embodiment, the output of the selector is passed through an analog processing circuit that can include one or more circuits selected from a group consisting of: a level shift circuit, a programmable offset circuit, and a programmable gain amplifier.\",\n \"In various aspects of the embodiment, the imaging device further comprises an analog processing selector that is operable to bypass one or more portions of the analog processing circuit.\",\n \"In other aspects, the imaging device further includes a comparator receiving an output from the analog to digital converter.\",\n \"Such a comparator is selected from a group consisting of: a programmable hardware comparator, a programmable software comparator, and a hardwired comparator.\",\n \"In some aspects of the embodiment, the pixel array includes at least one pseudo-pixel, and the imaging device additionally includes a pixel array selector that is operable to select the pseudo-pixel such that input derived from the pixel array is derived from the pseudo-pixel.\",\n \"Other aspects include a column amplifier that receives and amplifies the output from the pseudo-pixel.\",\n \"In some aspects, the reference input to the selector is a reset level that includes a pixel under reset condition, and the imaging device farther includes a circuit for selecting the reset level and pairing it with an output from the pixel array.\",\n \"In this way, the selector is operable to provide a differential pair consisting of the input derived from the pixel array and the reset level.\",\n \"In other aspects, the reference input to the selector is tied to a pad of the imaging device, and in yet other aspects, the reference input to the selector comprises one or more voltages generated internal to the imaging device.\",\n \"In other embodiments of the present invention, imaging devices and methods for using such are provided to detect and/or reduce image flicker.\",\n \"Such an imaging device includes an image sensor with one or more rows of pixels, where each of the pixels provides a pixel value.\",\n \"The device further includes a storage element and a summing circuit.\",\n \"Such a summing circuit can be a hardware summing circuit, either programmable or hardwired, or a software summing circuit including for example a programmable core.\",\n \"The summing circuit is operable to sum the pixel values for each of the one or more rows to create row sums, and to store the row sums to the storage element.\",\n \"In addition, the imaging device includes a programmable core that includes instructions executable by the programmable core to: determine a first energy value and a second energy value associated with the row sums; compare the first energy value and the second energy value to estimate a flicker frequency; and constrain an exposure duration associated with the image sensor to approximate a multiple of the flicker frequency.\",\n \"Such a programmable core can also be used for the summing circuit.\",\n \"In some aspects of the embodiment, the summing circuit is further operable to calculate row sums for a plurality of frames, where each of the plurality of frames includes pixel values from one or more rows of pixels, and where the instructions are further executable by the programmable core to: determine a first energy value and a second energy value associated with the row sums for each frame; calculate a variance between first energy values across the plurality of frames; calculate a variance between second energy values across the plurality of frames; and wherein the comparing the first energy value and the second energy value to estimate a flicker frequency comprises comparing the variance of the first energy values with the variance of the second energy values.\",\n \"In yet other aspects of the embodiment, the instructions are further executable by the programmable core to: compare the variance of the first energy values with the variance of the second energy values, and to compare the greater variance with a predefined threshold where constraining the exposure frequency is done when the greater variance is greater than the predefined threshold.\",\n \"In various aspects, a confidence counter is implemented to control switching from an exposure frequency to a multiple of an estimated flicker frequency.\",\n \"In such aspects, a change of frequencies can be limited to situations where the confidence counter changes sign.\",\n \"In one particular aspect of the embodiment, the summing circuit is further operable to calculate row sums for a plurality of frames, wherein each of the plurality of frames includes pixel values from the two or more rows of pixels.\",\n \"Further, the imaging device further includes an interrupt associated with the summing circuit, wherein assertion of the interrupt indicates that summing for the two or more rows of pixels for a frame is complete; and wherein the instructions are further executable by the programmable core to: receive the interrupt; determine a first energy value and a second energy value associated with the row sums for each frame; determine if a sufficient number of frames have been received to calculate a variance between first energy values and a variance between second energy values; calculate the variance between first energy values across the plurality of frames; calculate the variance between second energy values across the plurality of frames; and wherein the comparing the first energy value and the second energy value to estimate a flicker frequency comprises comparing the variance of the first energy values with the variance of the second energy values.\",\n \"Some aspects of the invention provide a method for automatically detecting flicker in an imaging device.\",\n \"The method includes providing an imaging device, wherein the imaging device includes an image sensor with two or more rows of pixels, and wherein each of the pixels provides a pixel value.\",\n \"In addition, the method includes retrieving at least a portion of the two or more rows of pixels, and summing the pixel values to create a row sum for each of the rows of pixels; and determining a first energy value and a second energy value associated with the row sums.\",\n \"The first and second energy values are compared to estimate a flicker frequency.\",\n \"Various embodiments of the present invention include a pixel with a charge evacuation mechanism for reducing noise evident at the output of the pixel.\",\n \"In one aspect, the pixel includes a reset element used in relation to charging a light sensitive element.\",\n \"The evacuation mechanism is used to dissipate charge build up about the reset element.\",\n \"Dissipation of the charge results in a reduction of noise at the output of the pixel.\",\n \"In particular embodiments, such a pixel and/or charge evacuation mechanism is implemented using complementary metal oxide semiconductor (“CMOS”), field effect transistor (“FET”) technology.\",\n \"Further, aspects of the present invention include systems and methods for utilizing such pixels.\",\n \"More particularly, such methods include approaches for applying various control signals to the pixel and systems include elements for producing the various control signals.\",\n \"One particular aspect of the invention provides a MOS pixel that includes a reset transistor with a reset drain and a reset gate.\",\n \"Further, the pixel includes a charge evacuation element that is electrically coupled to the reset drain and is operable to evacuate charge accumulated in a channel of the reset transistor.\",\n \"In some embodiments, the charge evacuation element is a MOS transistor with a charge evacuation drain, a charge evacuation source, and a charge evacuation gate.\",\n \"In particular cases, the charge evacuation source and the charge evacuation drain are both electrically coupled to the reset drain.\",\n \"In various embodiments, the pixel further includes a light sensitive element electrically coupled to the reset drain.\",\n \"Further embodiments comprise a source follower transistor that includes a source follower gate and a source follower drain, the source follower gate being electrically coupled to the reset drain.\",\n \"Such embodiments further include a selection transistor with a selection drain, a selection source, and a selection gate, where the selection source is electrically coupled to the source follower drain.\",\n \"A selection signal is electrically coupled to the source gate such that assertion thereof causes a representation of a signal from the node of the light sensitive element to be present on an output of the pixel.\",\n \"In some embodiments, the charge evacuation element is a MOS transistor with a charge evacuation source and a charge evacuation drain, both of which are coupled to the reset drain.\",\n \"The charge evacuation element further includes a charge evacuation gate.\",\n \"Further, such embodiments can include a reset signal driving the reset gate and a complement of the reset signal driving the charge evacuation gate.\",\n \"In some instances, the complement of the reset signal is delayed from the reset signal.\",\n \"Other embodiments of the present invention provide a circuit including a reset transistor comprising a reset gate and a reset drain.\",\n \"The circuit further includes a photodiode with a node of the photodiode electrically coupled to the reset drain, and a charge evacuation transistor that includes a charge evacuation gate, a charge evacuation drain, and a charge evacuation source.\",\n \"The charge evacuation source and the charge evacuation drain are both electrically coupled to the reset drain and the charge evacuation transistor is operable to evacuate charge accumulated in a channel of the reset transistor.\",\n \"Various embodiments of the circuit also include a source follower transistor comprised of a source follower gate and a source follower drain.\",\n \"The source follower gate is electrically coupled to the reset drain.\",\n \"the circuit further includes a selection transistor comprised of a selection source, a selection gate, and a selection drain.\",\n \"The selection source is electrically coupled to the source follower drain.\",\n \"A selection signal is provided that when asserted causes a representation of a signal from the photodiode to be present on an output of the pixel.\",\n \"Yet other embodiments of the present invention provide an imaging system.\",\n \"Such an imaging system includes a group of pixel elements.\",\n \"One or more of the pixel elements includes a reset element, a charge evacuation element, and a light sensitive element.\",\n \"In some cases, the imaging system further includes an optical device, wherein the optical device transfers light to the group of pixel elements, and wherein the light strikes the light sensitive element of the pixel elements.\",\n \"In addition, a timing circuit is included that provides at least one control signal to the group of pixel elements.\",\n \"In one particular instance, the group of pixel elements are arranged in a rectangular array.\",\n \"Yet another embodiment of the present invention provides an image sensor including a plurality of pixel devices.\",\n \"The plurality of pixel devices are arranged as a plurality of rows and a plurality of columns.\",\n \"Each of the pixel devices includes: a light detecting element, and a charge evacuation element for dissipating unwanted charge built up in the image sensor.\",\n \"Yet additional embodiments provide methods for detecting an image.\",\n \"Such methods include providing a pixel device that comprises a charge evacuation element, and applying a charge evacuation control signal to the charge evacuation element, wherein a charge accumulation in a channel of a reset transistor of the pixel device is reduced.\",\n \"Other embodiments provide systems and method for reducing power consumption in an imaging device.\",\n \"In one aspect, an imaging device is included with an image sensor and a sub-sample control system.\",\n \"The sub-sample control system is operable to identify one or more signals from the image sensor that are not processed.\",\n \"The imaging device further includes an analog processing circuit that receives the one or more signals from the image sensor at one or more inputs, and a processing control system that is operable to limit switching of the one or more inputs.\",\n \"In some aspects, the analog processing circuit comprises a column amplifier that receives the one or more inputs.\",\n \"In various aspects, the analog processing circuit comprises a level shift circuit, and/or a programmable or black offset circuit.\",\n \"In some aspects, the processing control system is operable to maintain a common mode feedback loop of an amplifier associated with the analog processing circuit in an active state, while limiting switching of the one or more inputs.\",\n \"In other aspects, the imaging device further includes a digital processing circuit, and the processing control system is further operable to disable the digital processing circuit.\",\n \"Yet other embodiments of the present invention provide a CMOS imager that includes an image sensor, and a parallel to serial data conversion unit.\",\n \"The parallel to serial data conversion unit receives an image derived from the image sensor and converts the image to a serial data stream.\",\n \"The serial data stream is output via a serial output interface.\",\n \"In some aspects of the embodiment, the image sensor comprises one or more pixels, and wherein the one or more pixels include a charge evacuation element.\",\n \"in other aspects, the serial output interface includes a clock signal and a data signal.\",\n \"The clock signal is only active when valid data is presented on the data signal.\",\n \"In yet other aspects, the serial output includes a clock signal, a data signal, and a qualifying signal.\",\n \"The qualifying signal indicates the presence of valid data on the data signal.\",\n \"These and other aspects are more fully developed in the detailed description below.\",\n \"Thus, the summary provides only a general outline of the embodiments according to the present invention.\",\n \"Many other objects, features and advantages of the present invention will become more fully apparent from the following detailed description, the appended claims and the accompanying drawings.\"\n ],\n [\n \"CROSS-REFERENCES TO RELATED APPLICATIONS This application claims the benefit of U.S.\",\n \"Provisional Patent Application No.\",\n \"60/294,388, filed on May 29, 2001, which is incorporated herein by reference in its entirety for all purposes.\",\n \"BACKGROUND OF THE INVENTION The present invention relates generally to systems and methods for detecting and/or transmitting images.\",\n \"More particularly, the present invention relates to detecting and transmitting images in relation to a cellular telephone.\",\n \"Image processors typically detect light reflected off of a subject, and convert the detected light to an electrical image.\",\n \"The electrical image is then sampled at a frequency that is the reciprocal of the integration time.\",\n \"In many cases, such a frequency is different from the frequency of light reflected off the subject.\",\n \"In such cases, a modulation of the displayed image can result.\",\n \"Such modulation appears as a flicker and can be very distracting.\",\n \"To overcome this problem, some image processors allow for manual selection between different integration times that allow a user to reduce the flicker.\",\n \"For example, where an image processor will be used in a 60 Hz and a 50 Hz lighting environment, a manual selection between an integration time associated with either 60 Hz or 50 Hz can be provided.\",\n \"This alleviates some flicker, but requires adjustment from a user.\",\n \"Furthermore, such an approach is limited to reducing flicker in a limited number of pre-determined environments, and thus may not be able to address various flicker situations.\",\n \"This is particularly problematic for mobile devices that are used in ever changing environments.\",\n \"Further, a limited reservoir of power typically exists for mobile devices.\",\n \"As such, power consuming imaging applications are often incompatible with mobile devices.\",\n \"Various approaches exist to increase the amount of power available to mobile devices, such that the power requirements of an imaging application are not prohibitive.\",\n \"However, such approaches often are both expensive and can result in increased dimensions of a mobile device.\",\n \"In some cases, light detected by an image processor is detected by a pixel array and a variety of analog processing circuitry.\",\n \"Output from the pixel array representing the amount of detected light is processed by analog circuitry to produce an image which is subsequently converted from the analog domain to the digital domain.\",\n \"The digital image is subsequently provided to a monitor of some sort for viewing.\",\n \"Testing such an imaging device can include reflecting light off a known image, and determining if the image was properly acquired and/or processed by the imaging device.\",\n \"This method provides an effective approach for testing such devices, however, the method is subject to a number of variables including lighting and image stability which can effect testing of the imaging device.\",\n \"Providing such a test, and controlling for such variables can be costly and time consuming.\",\n \"Further, such an approach tests the imaging device holistically, and is limited in its ability to identify sub-components of the imaging device which may have failed.\",\n \"Hence, for at least the aforementioned reasons, there exists a need in the art to provide advanced systems, methods and devices for detecting and/or transmitting images.\",\n \"BRIEF SUMMARY OF THE INVENTION The present invention provides systems, devices, and methods for imaging.\",\n \"More particularly, the present invention provides devices and methods for capturing images, formatting the images, and/or transmitting the images via a cellular telephone network or other wireless network.\",\n \"Various aspects of the present invention include an imaging device with one or more of the following units: a control unit, a serial interface unit, a parallel interface unit, an imaging unit, and a translation unit.\",\n \"In some embodiments, flicker detection and/or correction functionality is incorporated.\",\n \"Additionally, in some embodiments, a built-in self test associated with various analog circuitry in the imaging device is included.\",\n \"Further, in some embodiments, systems and methods for reducing power consumption by various analog circuitry within the imaging device are included.\",\n \"Yet further, in some embodiments, a parallel to serial conversion unit and associated serial output interface are included.\",\n \"These and many other novel aspects of the present invention are provided in greater detail in the proceeding portions of this document.\",\n \"One particular embodiment of the present invention provides an imaging device and methods for using such that provide for advanced testing ability.\",\n \"In one aspect, an imaging device includes a pixel array.\",\n \"The imaging device further includes a selector that is operable to select between an input derived from the pixel array and at least one reference input.\",\n \"Such selection between the input derived from the pixel array and the at least one reference input is based at least in part on a signal derived from a digital domain.\",\n \"Further, the imaging device includes an analog to digital converter that is operable to convert a signal derived from the selector to the digital domain.\",\n \"In some cases, such an embodiment provides an ability to verify the functionality of analog circuitry within the imaging device using a digital tester.\",\n \"In some aspects of the embodiment, the output of the selector is passed through an analog processing circuit that can include one or more circuits selected from a group consisting of: a level shift circuit, a programmable offset circuit, and a programmable gain amplifier.\",\n \"In various aspects of the embodiment, the imaging device further comprises an analog processing selector that is operable to bypass one or more portions of the analog processing circuit.\",\n \"In other aspects, the imaging device further includes a comparator receiving an output from the analog to digital converter.\",\n \"Such a comparator is selected from a group consisting of: a programmable hardware comparator, a programmable software comparator, and a hardwired comparator.\",\n \"In some aspects of the embodiment, the pixel array includes at least one pseudo-pixel, and the imaging device additionally includes a pixel array selector that is operable to select the pseudo-pixel such that input derived from the pixel array is derived from the pseudo-pixel.\",\n \"Other aspects include a column amplifier that receives and amplifies the output from the pseudo-pixel.\",\n \"In some aspects, the reference input to the selector is a reset level that includes a pixel under reset condition, and the imaging device farther includes a circuit for selecting the reset level and pairing it with an output from the pixel array.\",\n \"In this way, the selector is operable to provide a differential pair consisting of the input derived from the pixel array and the reset level.\",\n \"In other aspects, the reference input to the selector is tied to a pad of the imaging device, and in yet other aspects, the reference input to the selector comprises one or more voltages generated internal to the imaging device.\",\n \"In other embodiments of the present invention, imaging devices and methods for using such are provided to detect and/or reduce image flicker.\",\n \"Such an imaging device includes an image sensor with one or more rows of pixels, where each of the pixels provides a pixel value.\",\n \"The device further includes a storage element and a summing circuit.\",\n \"Such a summing circuit can be a hardware summing circuit, either programmable or hardwired, or a software summing circuit including for example a programmable core.\",\n \"The summing circuit is operable to sum the pixel values for each of the one or more rows to create row sums, and to store the row sums to the storage element.\",\n \"In addition, the imaging device includes a programmable core that includes instructions executable by the programmable core to: determine a first energy value and a second energy value associated with the row sums; compare the first energy value and the second energy value to estimate a flicker frequency; and constrain an exposure duration associated with the image sensor to approximate a multiple of the flicker frequency.\",\n \"Such a programmable core can also be used for the summing circuit.\",\n \"In some aspects of the embodiment, the summing circuit is further operable to calculate row sums for a plurality of frames, where each of the plurality of frames includes pixel values from one or more rows of pixels, and where the instructions are further executable by the programmable core to: determine a first energy value and a second energy value associated with the row sums for each frame; calculate a variance between first energy values across the plurality of frames; calculate a variance between second energy values across the plurality of frames; and wherein the comparing the first energy value and the second energy value to estimate a flicker frequency comprises comparing the variance of the first energy values with the variance of the second energy values.\",\n \"In yet other aspects of the embodiment, the instructions are further executable by the programmable core to: compare the variance of the first energy values with the variance of the second energy values, and to compare the greater variance with a predefined threshold where constraining the exposure frequency is done when the greater variance is greater than the predefined threshold.\",\n \"In various aspects, a confidence counter is implemented to control switching from an exposure frequency to a multiple of an estimated flicker frequency.\",\n \"In such aspects, a change of frequencies can be limited to situations where the confidence counter changes sign.\",\n \"In one particular aspect of the embodiment, the summing circuit is further operable to calculate row sums for a plurality of frames, wherein each of the plurality of frames includes pixel values from the two or more rows of pixels.\",\n \"Further, the imaging device further includes an interrupt associated with the summing circuit, wherein assertion of the interrupt indicates that summing for the two or more rows of pixels for a frame is complete; and wherein the instructions are further executable by the programmable core to: receive the interrupt; determine a first energy value and a second energy value associated with the row sums for each frame; determine if a sufficient number of frames have been received to calculate a variance between first energy values and a variance between second energy values; calculate the variance between first energy values across the plurality of frames; calculate the variance between second energy values across the plurality of frames; and wherein the comparing the first energy value and the second energy value to estimate a flicker frequency comprises comparing the variance of the first energy values with the variance of the second energy values.\",\n \"Some aspects of the invention provide a method for automatically detecting flicker in an imaging device.\",\n \"The method includes providing an imaging device, wherein the imaging device includes an image sensor with two or more rows of pixels, and wherein each of the pixels provides a pixel value.\",\n \"In addition, the method includes retrieving at least a portion of the two or more rows of pixels, and summing the pixel values to create a row sum for each of the rows of pixels; and determining a first energy value and a second energy value associated with the row sums.\",\n \"The first and second energy values are compared to estimate a flicker frequency.\",\n \"Various embodiments of the present invention include a pixel with a charge evacuation mechanism for reducing noise evident at the output of the pixel.\",\n \"In one aspect, the pixel includes a reset element used in relation to charging a light sensitive element.\",\n \"The evacuation mechanism is used to dissipate charge build up about the reset element.\",\n \"Dissipation of the charge results in a reduction of noise at the output of the pixel.\",\n \"In particular embodiments, such a pixel and/or charge evacuation mechanism is implemented using complementary metal oxide semiconductor (“CMOS”), field effect transistor (“FET”) technology.\",\n \"Further, aspects of the present invention include systems and methods for utilizing such pixels.\",\n \"More particularly, such methods include approaches for applying various control signals to the pixel and systems include elements for producing the various control signals.\",\n \"One particular aspect of the invention provides a MOS pixel that includes a reset transistor with a reset drain and a reset gate.\",\n \"Further, the pixel includes a charge evacuation element that is electrically coupled to the reset drain and is operable to evacuate charge accumulated in a channel of the reset transistor.\",\n \"In some embodiments, the charge evacuation element is a MOS transistor with a charge evacuation drain, a charge evacuation source, and a charge evacuation gate.\",\n \"In particular cases, the charge evacuation source and the charge evacuation drain are both electrically coupled to the reset drain.\",\n \"In various embodiments, the pixel further includes a light sensitive element electrically coupled to the reset drain.\",\n \"Further embodiments comprise a source follower transistor that includes a source follower gate and a source follower drain, the source follower gate being electrically coupled to the reset drain.\",\n \"Such embodiments further include a selection transistor with a selection drain, a selection source, and a selection gate, where the selection source is electrically coupled to the source follower drain.\",\n \"A selection signal is electrically coupled to the source gate such that assertion thereof causes a representation of a signal from the node of the light sensitive element to be present on an output of the pixel.\",\n \"In some embodiments, the charge evacuation element is a MOS transistor with a charge evacuation source and a charge evacuation drain, both of which are coupled to the reset drain.\",\n \"The charge evacuation element further includes a charge evacuation gate.\",\n \"Further, such embodiments can include a reset signal driving the reset gate and a complement of the reset signal driving the charge evacuation gate.\",\n \"In some instances, the complement of the reset signal is delayed from the reset signal.\",\n \"Other embodiments of the present invention provide a circuit including a reset transistor comprising a reset gate and a reset drain.\",\n \"The circuit further includes a photodiode with a node of the photodiode electrically coupled to the reset drain, and a charge evacuation transistor that includes a charge evacuation gate, a charge evacuation drain, and a charge evacuation source.\",\n \"The charge evacuation source and the charge evacuation drain are both electrically coupled to the reset drain and the charge evacuation transistor is operable to evacuate charge accumulated in a channel of the reset transistor.\",\n \"Various embodiments of the circuit also include a source follower transistor comprised of a source follower gate and a source follower drain.\",\n \"The source follower gate is electrically coupled to the reset drain.\",\n \"the circuit further includes a selection transistor comprised of a selection source, a selection gate, and a selection drain.\",\n \"The selection source is electrically coupled to the source follower drain.\",\n \"A selection signal is provided that when asserted causes a representation of a signal from the photodiode to be present on an output of the pixel.\",\n \"Yet other embodiments of the present invention provide an imaging system.\",\n \"Such an imaging system includes a group of pixel elements.\",\n \"One or more of the pixel elements includes a reset element, a charge evacuation element, and a light sensitive element.\",\n \"In some cases, the imaging system further includes an optical device, wherein the optical device transfers light to the group of pixel elements, and wherein the light strikes the light sensitive element of the pixel elements.\",\n \"In addition, a timing circuit is included that provides at least one control signal to the group of pixel elements.\",\n \"In one particular instance, the group of pixel elements are arranged in a rectangular array.\",\n \"Yet another embodiment of the present invention provides an image sensor including a plurality of pixel devices.\",\n \"The plurality of pixel devices are arranged as a plurality of rows and a plurality of columns.\",\n \"Each of the pixel devices includes: a light detecting element, and a charge evacuation element for dissipating unwanted charge built up in the image sensor.\",\n \"Yet additional embodiments provide methods for detecting an image.\",\n \"Such methods include providing a pixel device that comprises a charge evacuation element, and applying a charge evacuation control signal to the charge evacuation element, wherein a charge accumulation in a channel of a reset transistor of the pixel device is reduced.\",\n \"Other embodiments provide systems and method for reducing power consumption in an imaging device.\",\n \"In one aspect, an imaging device is included with an image sensor and a sub-sample control system.\",\n \"The sub-sample control system is operable to identify one or more signals from the image sensor that are not processed.\",\n \"The imaging device further includes an analog processing circuit that receives the one or more signals from the image sensor at one or more inputs, and a processing control system that is operable to limit switching of the one or more inputs.\",\n \"In some aspects, the analog processing circuit comprises a column amplifier that receives the one or more inputs.\",\n \"In various aspects, the analog processing circuit comprises a level shift circuit, and/or a programmable or black offset circuit.\",\n \"In some aspects, the processing control system is operable to maintain a common mode feedback loop of an amplifier associated with the analog processing circuit in an active state, while limiting switching of the one or more inputs.\",\n \"In other aspects, the imaging device further includes a digital processing circuit, and the processing control system is further operable to disable the digital processing circuit.\",\n \"Yet other embodiments of the present invention provide a CMOS imager that includes an image sensor, and a parallel to serial data conversion unit.\",\n \"The parallel to serial data conversion unit receives an image derived from the image sensor and converts the image to a serial data stream.\",\n \"The serial data stream is output via a serial output interface.\",\n \"In some aspects of the embodiment, the image sensor comprises one or more pixels, and wherein the one or more pixels include a charge evacuation element.\",\n \"in other aspects, the serial output interface includes a clock signal and a data signal.\",\n \"The clock signal is only active when valid data is presented on the data signal.\",\n \"In yet other aspects, the serial output includes a clock signal, a data signal, and a qualifying signal.\",\n \"The qualifying signal indicates the presence of valid data on the data signal.\",\n \"These and other aspects are more fully developed in the detailed description below.\",\n \"Thus, the summary provides only a general outline of the embodiments according to the present invention.\",\n \"Many other objects, features and advantages of the present invention will become more fully apparent from the following detailed description, the appended claims and the accompanying drawings.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS A further understanding of the nature and advantages of the present invention may be realized by reference to the figures which are described in remaining portions of the specification.\",\n \"In the figures, like reference numerals are used throughout several figures to refer to similar components.\",\n \"In some instances, a sub-label consisting of a lower case letter is associated with a reference numeral to denote one of multiple similar components.\",\n \"When reference is made to a reference numeral without specification to an existing sub-label, it is intended to refer to all such multiple similar components.\",\n \"FIG.\",\n \"1 is a block diagram of a CMOS imager in accordance with various embodiments of the present invention; FIG.\",\n \"2 is a timing diagram of frame rate control implemented in the CMOS imager of FIG.\",\n \"1; FIG.\",\n \"3 is a timing diagram illustrating the various clocks used in relation to the CMOS imager of FIG.\",\n \"1; FIG.\",\n \"4 is a block diagram of a sensor in accordance with one embodiment of the present invention; FIG.\",\n \"5 is a block diagram of a virtual image window in accordance with various embodiments of the present invention; FIG.\",\n \"6 illustrates a Bayer grid array; FIG.\",\n \"7 illustrates a pixel circuit in accordance with various embodiments of the present invention; FIG.\",\n \"8 is a flow diagram illustrating a method for using the pixel circuit of FIG.\",\n \"7 in accordance with an embodiment of the present invention; FIG.\",\n \"9 are timing diagrams illustrating the operation of the pixel of FIG.\",\n \"7.FIG.\",\n \"10 is an exemplary column amplifier circuit; FIGS.\",\n \"11-13 are timing diagrams illustrating the operation of the column amplifier circuit of FIG.\",\n \"10; FIG.\",\n \"14 is a timing diagram illustrating the exposure and sensing of a flame of video in accordance with an embodiment of the present invention; FIG.\",\n \"15 is a timing diagram illustrating sampling of a pixel array incorporated in the sensor of FIG.\",\n \"4; FIG.\",\n \"16 illustrates high gain and high dynamic range modes of the level shift circuit of FIG.\",\n \"4; FIG.\",\n \"17 illustrates the use of black level and gain controls in relation to the sensor of FIG.\",\n \"4; FIG.\",\n \"18 is a timing diagram of various control signals associated with the sensor of FIG.\",\n \"4; FIG.\",\n \"19 is a timing diagram illustrating the interaction of various standby mode control signals; FIG.\",\n \"20 is a block diagram of a standby mode circuit in accordance with embodiments of the present invention; FIG.\",\n \"21 illustrates an implementation of a standby mode in a switched capacitor circuit in accordance with embodiments of the present invention; FIG.\",\n \"22 is a flow diagram illustrating a method of entering and exiting a standby mode in accordance with embodiments of the present invention; FIG.\",\n \"23 is a block diagram of a power distribution circuit for use in relation to the CMOS imager of FIG.\",\n \"1; FIG.\",\n \"24 is a block diagram of a translation unit for use in relation to the CMOS imager of FIG.\",\n \"1; FIG.\",\n \"25 illustrates the placement of color components by the translation unit of FIG.\",\n \"24; FIG.\",\n \"26 is a block diagram of a flicker detection and/or correction block in accordance with embodiments of the present invention; FIG.\",\n \"27 is a graph illustrating an exemplary row average across a frame of data; FIG.\",\n \"28 is a flow diagram of a method for detecting and/or correcting flicker in accordance with embodiments of the present invention; FIG.\",\n \"29 is a timing/block diagram illustrating the operation of qualification signals used in relation to providing parallel output data from the CMOS imager of FIG.\",\n \"1; FIG.\",\n \"30 is a timing diagram illustrating vertical and horizontal qualification signals used in relation to providing parallel output data from the CMOS imager of FIG.\",\n \"1; FIG.\",\n \"31 is a timing diagram illustrating the output of Bayer data via a parallel output interface; FIG.\",\n \"32 is a timing diagram illustrating the output of REV data via a parallel output interface; FIG.\",\n \"33 is a timing diagram illustrating frame dropping; FIG.\",\n \"34 is a timing diagram illustrating unqualified serial output in accordance with embodiments of the present invention; and FIG.\",\n \"35 is a timing diagram illustrating qualified serial output in accordance with embodiments of the present invention.\",\n \"DETAILED DESCRIPTION OF THE INVENTION The present invention provides systems, devices, and methods for imaging.\",\n \"More particularly, the present invention provides devices and methods for capturing images, formatting the images, and/or transmitting the images via a cellular telephone network or other wireless network.\",\n \"In one particular embodiment of the present invention, an imaging device including a CMOS sensor is provided.\",\n \"The imaging device includes an imager array, sensor timing control, and image processing algorithms integrated onto a single chip.\",\n \"Such an imaging device can produce CIF (i.e., 352×288) resolution image for still capture, and QCIF (i.e., 176×144) resolution image for video applications.\",\n \"In addition, the imaging device can perform various correction algorithms on a captured image to create a corrected YCrCb (4:2:2) image.\",\n \"In an embodiment, the imager array is 384×316 physical pixels including 376×296 active pixels.\",\n \"The pixels can be mapped to a configurable size of output Bayer grid array (e.g., windowing).\",\n \"Pixel size can be 6.4 μm×6.4 μm.\",\n \"QCIF resolution can be created by either or both of down-sampling and filtering.\",\n \"In some embodiments, a resulting Bayer grid (raw RGB output) or a recreated YCrCb image is sent out on a digital ten (10) bit parallel interface.\",\n \"The ten (10) bit parallel interface provides access to a captured video image on a line by line basis.\",\n \"In addition, an output clock and other qualifying signals are provided in association with the ten (10) bit parallel interface.\",\n \"The imaging device can include electronic exposure control from 375 nsec up to a full frame (e.g., 33.33 msec at a maximum frame rate of thirty frames per second).\",\n \"Further, automatic dark level offset, a separate gain for each of the color components, and configurable offset for dynamic range stretching, as well as, image enhancement algorithms can be provided.\",\n \"Such image enhancement algorithms can include black level offset, fault pixel correction, color interpolation, color and white balance correction, gamma correction, and/or auto exposure and auto white balance algorithms.\",\n \"In addition, noise reduction processes can be applied to reduce noise in general, and fixed pattern noise (“FPN”) in particular.\",\n \"The imaging device can operate on a single 2.8 volt power supply and a reduced-power mode can be implemented to reduce power consumption when full operation of the imaging device is not necessary.\",\n \"In addition, the imaging device can include a sleep mode for further reducing power consumption.\",\n \"In some embodiments, the imaging device is designed for use under the control of an external processor, which in some cases is a Digital Signal Processor.\",\n \"The external processor can act as a system host (“Host”).\",\n \"As such, the external processor controls the mode of the imaging device, monitors system status, and compresses an output image stream.\",\n \"For example, the external processor can control the frame-rate, the size of the requested image, and several registers for image processing, such as gamma-correction and gain registers.\",\n \"Such control can be provided via a programmable core integrated with the imaging device.\",\n \"Block Diagram FIG.\",\n \"1 illustrates a block diagram of an imaging device 100 in accordance with embodiments of the present invention.\",\n \"Imaging device 100 includes a serial interface unit 106, a control unit 110, an imaging unit 130, a translation unit 150, a testability unit 170, and a parallel interface unit 190.The various units are electrically coupled via a control bus 101, an image bus 131, a translation bus 151, a parallel interface bus 191, a UART bus 120, a DeICE bus 125, and a serial bus 107.Inputs and outputs to/from imaging device 100 can be provided via an I2C input/output (“I/O”) 108, a UART I/O 121, a DeICE I/O 126, and/or a parallel video interface 192.In addition, imaging device 100 includes a clock unit 102 with a reset 171 and a clock 172 input.\",\n \"Serial Interface In one embodiment, serial I/O 108 includes a clock and data input.\",\n \"Such a serial interface can be used to program various registers of imaging device 100.Various different serial interface types that are known in the art can be incorporated into imaging device 100 in accordance with the present invention.\",\n \"Control Unit Control unit 110 implements parameters used in relation to the different units within imaging device 100, and calculates various parameters for image enhancement.\",\n \"In some embodiments, control unit 110 includes a programmable processor core 112, and in one particular embodiment, programmable processor core 112 is a S0186Turbo processor core.\",\n \"Control unit 110 also includes a core memory 118, and interface hardware (not shown) to UART I/O 121 and DeICE 126 I/O.\",\n \"Further, control unit 110 includes an arbitrator 116 for controlling access to control bus 101, and a frame-rate mechanism 114 for controlling the frame rate output video images from imaging device 100.Via arbitrator 116, programmable processing core 112, serial I/O 108, UART I/O 121, and/or DeICE I/O 126 can access registers maintained on imaging device 100 over control bus 101.In one particular embodiment, when programmable processing core 112 is disabled by, for example, entering a clock disable mode, serial I/O 108 is granted immediate access to control bus 101 without adhering to an arbitration scheme implemented by arbitrator 116.Such a clock disable mode is discussed in more detail with reference to clock unit 102.For purposes of the immediate discussion, clock unit 102 produces a control line that indicates if programmable processing core 112 is active, or if it is held in reset mode or has its clock shut down.\",\n \"Alternatively, when programmable processing core 112 is active, arbitrator 116 grants access to control bus 101 according to a determined priority scheme.\",\n \"One such priority scheme is described in the 80186Turbo reference manual.\",\n \"Programmable processor core 112 serves as the main control source for imaging device 100, controlling other units via control bus 101.Such control can be provided by manipulating address, data and read/write signals associated with control bus 101.As just one example, programmable processing core 112 sets up the working mode of imaging device 100 according to the parameters that are received from a host (not shown) via serial I/O 108, UART I/O 121, and/or DeICE I/O 126.In addition, programmable processing core 112 can regulate the operation of the various units within imaging device 100.In one particular embodiment, programmable processor core 112 supports a twenty-bit address memory space for data and program memories, and a fourteen-bit address spaced for I/O mapped peripherals.\",\n \"In other embodiments, only a twelve-bit memory space and an eight-bit I/O space (program and data) are supported.\",\n \"Programs controlling the operation of imaging device 100 can be loaded to core memory 118 via serial I/O 108, and/or via UART I/O 121.Once the program is loaded, control unit 110 calculates and updates various image processing parameters according to a selected working mode.\",\n \"After a complete frame is read from imaging unit 130 and processed by translation unit 150, control unit 110 reads statistical results of the frame, and produces the exposure, gain and auto-white balance parameters for the next frame.\",\n \"In this way, control unit 110 continuously monitors operation of imaging device 100 and modifies such operation to assure that it conforms to the downloaded program.\",\n \"In one particular embodiment, various parameters associated with the operation of imaging device 100 are written to registers associated with the various units within imaging device 100.Such a process can be performed by writing the parameters to core memory 118, after which programmable processing core 112 writes the values to registers maintained within the various units to which the parameters pertain.\",\n \"Alternatively, such parameters can be written directly to the various units by the Host.\",\n \"To facilitate such processes, imaging unit 130, translation unit 150, and parallel interface unit 190 can include double buffered registers so that parameters can be loaded into such registers at any time without interfering with continuing processes.\",\n \"Imaging device 100 operates in a default standby mode, with sensor 132 turned off and translation unit 150 and parallel interface unit 190 inactive.\",\n \"Once the program is loaded, control unit 110 awaits instructions from the Host.\",\n \"The Host can pass commands to control unit 110 by writing to memory 118 at HOST-FIFO address space.\",\n \"After writing data to the HOST-FIFO address space, the Host can activate an interrupt to programmable processing core 112.Programmable processing core 112 then reads the data and responds according to the instructions.\",\n \"Once the frame counters are active, a vertical synchronization interrupt signal (“VSYNC”) is asserted every time a frame boundary for an image is detected.\",\n \"VSYNC is used by programmable processing core 112 to govern software frame-rate control, or by frame-rate mechanism 114 to govern hardware frame-rate control.\",\n \"When an image is being processed by imaging device 100, statistics are collected in translation unit 150.At the end of a frame processing, translation unit 150 activates an interrupt signal, END_TRANSLATION, indicating that programmable processing core 112 can read the statistics from translation unit 150.Programmable processing core 112 utilizes these statistics to calculate exposure, gain and color processing parameters for the next frame.\",\n \"In some embodiments, such calculations are provided in accordance with auto-exposure, auto-white-balance, and/or dynamic range algorithms.\",\n \"The calculations result in a new set of parameters that are written by control unit 110 to registers maintained in association with imaging unit 130 and translation unit 150 before the following VSYNC.\",\n \"Further, control unit 110 can be interrupted by either or both of UART I/O 121 and serial I/O 108.Such interrupts are generally processed as soon as programmable processing core 112 becomes available.\",\n \"Reasonable response time to such interrupts helps assure that the I/O pipelines do not become overfilled.\",\n \"The following Table 1 lists a series of tasks completed as imaging device 100 is programmed and started.\",\n \"TABLE 1 Imaging Device Tasks EVENT Task Comments Reset Programmable processing core 112 is frozen Deassert Reset Execute boot program - check configuration register.\",\n \"If boot from UART I/O 121, wait for UART message.\",\n \"If from serial I/O 108, program is loaded, and can subsequently be executed.\",\n \"According to program, initiate registers and activate imager frame counters.\",\n \"VSYNC Asserts Interrupt processes.\",\n \"Activate translation unit 150 First frame exposure has for next frame.\",\n \"Execute started, no image is being frame-rate control routine.\",\n \"read out.\",\n \"VSYNC Interrupt processes.\",\n \"Asserts Execute frame-rate control Image is being sent from routine imaging unit 130 to translation unit 150 in accordance with a frame-rate control.\",\n \"END_TRANSLATION Interrupt processes.\",\n \"Asserts Read statistics results from The algorithm defines the translation unit 150.sequence of the calculations.\",\n \"Calculate gain and offset for In some instances, not all imaging unit 130.Calculate calculations are performed exposure parameters, white for every frame.\",\n \"balance correction, and color correction matrix.\",\n \"Write calculated parameters to various units.\",\n \"VSYNC Asserts Interrupt processes.\",\n \"Execute frame-rate control routine.\",\n \"In one embodiment, programmable processing core 112 includes six interrupts.\",\n \"Such interrupts can include, but are not limited to, two software timer interrupts used for timeout purposes, and interrupts associated with UART I/O 121 and serial I/O 108.Additionally, VSYNC and END_TRANSLATION are provided.\",\n \"As previously discussed, VSYNC indicates a frame boundary has been detected, and END_TRANSLATION indicates that translation unit 150 has completed processing a frame.\",\n \"Upon assertion of END_TRANSLATION, translation has gathered and/or calculated various statistics associated with the processed frame that can now be read and utilized by programmable processing core 112.Programmable processing core 112 utilizes the statistics to perform one or more of the following algorithms: auto exposure (e.g., exposure parameters calculation), auto white balance (e.g., white balance coefficients calculation and/or full color matrix calculation), and dynamic range stretching, (e.g., gain and offset calculations).\",\n \"Thus, for example, if the auto-exposure and auto-white-Balance options are enabled, the parameters calculated by programmable processing core 112 in relation to such controls are passed to imaging unit 130 where they are used to control exposure and processing of a subsequent image frame.\",\n \"In addition, programmable processing core 112 can turn its own clock off by writing to a disable clock register maintained in clock unit 102.In some embodiments, the clock is turned back on whenever any activity is detected on a receive pin associated with UART I/O 121, or if any interrupt signal is activated.\",\n \"Disabling programmable processing core 112 helps to conserve power.\",\n \"In addition to controlling the rate of image output via parallel video interface 192, control unit 110 also controls the frame-rate associated with image processing internal to imaging device 100.In an embodiment, three registers are used to effectuate frame rate control.\",\n \"The first, Frame_rate_en, holds an enable bit for a frame-rate mechanism 114.When frame-rate mechanism 114 is disabled, the array content is read according to a second register, Frame_rate_read, and the data is sent out on parallel interface 192.When frame-rate mechanism 114 is activated, two eight-bit registers define whether a particular frame is to be read out, or dropped.\",\n \"When a frame is to be output via parallel video interface 192, a control signal, c_readframe, which is synchronized to a frame boundary signal, Im_Frame 910, is sent to imaging unit 130 to enable output of the frame data to translation unit 150.In addition, a control signal, C_Outframe, which is also synchronized to Im_Frame 910 signal is sent to parallel interface unit 190 to enable the data received from translation unit 150 to be output via parallel video interface 192 at a rate defined by the Frame_rate_send register value.\",\n \"As illustrated in FIG.\",\n \"2, when c_readframe is asserted high, the content of a pixel array within sensor 132 is transferred from imaging unit 130 to translation unit 150.When C_Outframe is asserted high, the data is sent out on parallel video interface 192.In some embodiments, when C_Outframe is deasserted, the output data is ignored by parallel interface unit 190, but the output pins are driven.\",\n \"While the data lines of parallel video interface 192 are driven, the vertical and horizontal synchronization signals associated with the data are not driven.\",\n \"In other embodiments, when C_Outframe is deasserted, the output data is ignored by parallel interface unit 190, and the output pins are not driven to conserve power and reduce noise.\",\n \"Clock Unit Clock unit 102 receives clock input 172 signal and reset input 171.Based on these signals, as well as control signals from control unit 110, clock unit 102 produces the various clock and reset signals to the different units within imaging device 100.In addition, clock unit 102 samples the configuration pins while reset signal 171 is asserted, and produces several configuration signals based upon the sensed configuration arrangement.\",\n \"The configuration inputs determine the working clock rate, the type of serial interface (SSI or I2C compatible) that is operational on serial I/O 108 and the program-loading source (serial interface or UART) that is operational on UART I/O 121.Further, clock unit 102 produces a control line that indicates if programmable processing core 112 is active, or if it is held in reset mode or has its clock shut down.\",\n \"As previously discussed, this signal enables the bypass of arbitrator 116 for accessing control bus 101.Clock unit 102 produces the device's general reset signal.\",\n \"An internal reset signal, Reset_i 104, is activated when reset input 171 is asserted, by the setting of a software reset register, or by change in phase locked loop configuration.\",\n \"Clock unit 102 also synchronizes Reset_i 104 to an internal clock signal, Rclk 103.Clock unit 102 also produces a signal that resets programmable processing core 112.This reset signal is activated when Reset_i 104 is active and/or by a special reset command from serial interface unit 106.When the configuration of imaging device 100 calls for program loading through serial interface unit 106, programmable processing core 112 is held in reset state by this signal until released by a load complete command.\",\n \"For power reduction purposes, in some embodiments clock signals from clock unit 102 may be turned off.\",\n \"This can be accomplished under direction from either programmable processing core 112 or the Host.\",\n \"As previously discussed, the clocks can be restored whenever an interrupt signal is detected.\",\n \"Alternatively, clocks going to units other than control unit 110 can be turned off, and subsequently restored by issuing a command from programmable processing core 112.In an embodiment, clock unit 102 produces the following signals: an Rclk 601, an Aclk 603, a Reg_clk 609, an Rc_pix signal 605, an Rclkc, an Rclky, and an Rclkp.\",\n \"Rclk 601 is the main internal clock signal generated from clock input 172.A division factor is applied to clock input 172 to produce Rclk 601, such division factor being determined by reading the configuration pins when reset input 171 is asserted.\",\n \"Aclk 603 is a clock signal to the analog part of imaging unit 130 and is synchronous to Rclk 601, with a one-half Rclk 601 cycle skew between them, at the rate of the pixel-cycles.\",\n \"Thus, the positive going edge of Aclk 603 follows every fourth positive edge of Rclk 601.Reg_clk 609 is a clock signal to the voltage regulators.\",\n \"It is one-sixteenth the rate of Rclk 601.The rising edge of Reg_clk 609 coincides with the rising edge of Rclk 601, when Rc_pix 605 is asserted.\",\n \"Rc_pix 605 is a signal in the digital domain that marks the Rclk 601 cycle that coincides with the first low cycle of Aclk 603.The first positive going clock edge after an indication that a phase lock loop is ready is qualified as Rc_pix 605.Rclkc is a clock signal to control unit 110 that is identical to Rclk 601.Similarly, Rclky is identical to Rclk 601 and is the clock signal to translation unit 150.Rclkp is a clock signal to parallel interface 190 that is derived from clock input 172.Rclkp can either be identical in frequency to clock input 172, or divided according to a clock division register, Rclkp_config.\",\n \"Rclkp is used for serial output mode and is typically eight to sixteen times the rate of a pixel-clock when serial output is required.\",\n \"Rclk_edge 607 is a qualifying signal to parallel interface unit 190 and is used for serial output mode.\",\n \"FIG.\",\n \"3 is an exemplary timing diagram 600 illustrating one example of the timing of the various signals generated by clock unit 102.The operating mode of imaging device 100 determines the clock configuration.\",\n \"The following Table 2 lists the different operating modes, and the clocking scheme that is associated with each of the modes: TABLE 2 Clock Modes Input Supported Rclk divide Rclkp frequency fps Supported output format factor divide factor 8 MHz 15 fps Bayer, YUV parallel (CIF, QCIF) 1 NA Bayer 1x serial - QCIF 1 1 12 MHz 20 fps Bayer, YUV parallel (CIF, QCIF) 1 NA Bayer 1x serial - QCIF 1 1 16 MHz 30 fps Bayer, YUV parallel (CIF, QCIF) 1 NA Bayer 1x serial - QCIF 1 1 15 fps Bayer, YUV parallel (CIF, QCIF) 2 NA Bayer 1x serial - QCIF 2 2 Bayer 1x serial - CIF 2 1 YUV 1x serial - QCIF 2 1 Bayer 2x serial - QCIF 2 1 32 MHz 30 fps Bayer, YUV parallel (CIF, QCIF) 2 NA Bayer 1x serial - QCIF 2 2 Bayer 1x serial - CIF 2 1 YUV 1x serial - QCIF 2 1 Bayer 2x serial - QCIF 2 1 15 fps Bayer, YUV parallel (CIF, QCIF) 4 NA Bayer 1x serial - QCIF 4 4 Bayer 1x serial - CIF 4 2 YUV 1x serial - QCIF 4 2 YUV 1x serial - CIF 4 1 Bayer 2x serial - QCIF 4 2 Bayer 2x serial - CIF 4 1 YUV 2x serial - QCIF 4 1 64 MHz 30 fps Bayer, YUV parallel (CIF, QCIF) 4 NA Bayer 1x serial - QCIF 4 4 Bayer 1x serial - CIF 4 2 YUV 1x serial - QCIF 4 2 YUV 1x serial - CIF 4 1 Bayer 2x serial - QCIF 4 2 Bayer 2x serial - CIF 4 1 YUV 2x serial - QCIF 4 1 15 fps Bayer, YUV parallel (CIF, QCIF) 8 NA Bayer 1x serial - QCIF 8 8 Bayer 1x serial - CIF 8 4 YUV 1x serial - QCIF 8 4 YUV 1x serial - CIF 8 2 Bayer 2x serial - QCIF 8 4 Bayer 2x serial - CIF 8 2 YUV 2x serial - QCIF 8 2 YUV 2x serial - CIF 8 1 Various combinations of frequencies for the input clock, Rclk and Rclkp are summarized in Table 3 below: TABLE 3 Various Input and Output Clock Frequencies clock input 172 Rclk dividing factor Rclkp dividing factor) 8 MHz, 12 MHz, 16 MHz 1 1 16 MHz, 32 MHz 2 1 2 2 32 MHz, 64 MHz 4 1 4 2 4 4 64 MHz 8 1 8 2 8 4 8 8 Imaging Unit Imaging unit 130 provides the central functionality of imaging device 100.Imaging unit 130 includes a timing control and address generator unit (“TCAG”) 136 that receives timing signals from control unit 110 and produces the sequence signals to sensor 132 and ADC 134 for integration (e.g., exposure control), read and reset operations.\",\n \"Sensor 132 includes a CIF size pixel array, level shift circuitry, gain control circuitry, programmable or black offset circuitry, gamma-correction circuitry and readout circuits.\",\n \"FIG.\",\n \"4 illustrates a block diagram 792 of the analog processing portion of imaging unit 130 including sensor 132 and ADC 134.Sensor 132 includes a pixel array 700, a column amplifier 703, an analog multiplexer (“MUX”) 709, a source selector mux 713, a level shift block 719, a black offset block 723, a first stage programmable gain amplifier 729, and a second stage programmable gain amplifier 733.In addition, sensor 132 includes a column logic block 739 and a reset level block 743.It should be noted that column amplifier 703 represents a vector of column amplifiers.\",\n \"Column logic within imaging device 100 can be designed such that only two of the 384 total column amplifiers within column amplifier 703 are active at any given time.\",\n \"FIG.\",\n \"5, illustrates one embodiment of pixel array 700 associated with sensor 132.Pixel array 700 includes an array of 376 by 296 active pixels 720 surrounded by four black column is 730 on each side.\",\n \"It is also surrounded by sixteen black lines 710 above and four black lines below the active image array, totaling 384 by 316.Each pixel in pixel array 700 is covered by a color filter, thus creating a Bayer grid format 800 as illustrated in FIG.\",\n \"6.Referring again to FIG.\",\n \"4, sensor 132 receives a number of timing and address signals from TCAG 136.More particularly, pixel array 700 receives the decoded and time-shaped line reset, Im_line_rst[328:0] (element 707), and line select, Im_line_sel[328:0] (element 727), signals.\",\n \"In some embodiments of imaging device 100, each pixel within pixel array 700 includes a charge injection compensation circuit.\",\n \"Referring to FIG.\",\n \"7, a pixel 3000 is illustrated in accordance with embodiments of the present invention that include such a charge injection compensation circuit.\",\n \"Pixel 3000 includes a reset element 3350, a source follower transistor 3360, a selection element 3370, a charge evacuation element 3380, and a light sensitive element 3390.The inputs to pixel 3000 include a reset control signal (im_line_rst 707), a selection control signal im_line_sel 727), and a charge evacuation control signal (im_line_rst_n 717).\",\n \"Further, pixel 3000 provides an output signal 742.In an embodiment, pixel 3000 is attached to a voltage source 3345 and a common ground 3355.Source follower transistor 3360 can be any of a number of transistor types.\",\n \"In one particular embodiment, source follower transistor 3360 is a CMOS FET.\",\n \"Source follower transistor 3360 includes a source follower gate 3361, a source follower source 3362 and a source follower drain 3363.Each of evacuation element 3380, reset element 3350, and photodiode 3390 are electrically connected to source follower gate 3361.For purposes of this document, charge evacuation element 3380 can be any device capable of dissipating charge from pixel 3000.Thus, for example, charge evacuation element 3380 can be a switch, a FET, a bipolar junction transistor (“BJT”), a switched capacitor, or any other such device.\",\n \"In one particular embodiment of the present invention, charge evacuation element 3380 is CMOS transistor as illustrated in FIG.\",\n \"7.In some cases, such CMOS transistors can be comprised of N-FET devices, P-FET devices, and/or combinations thereof as known in the art.\",\n \"In such an embodiment, charge evacuation element 3380 includes a charge evacuation gate 3381, a charge evacuation drain 3383, and a charge evacuation source 3382.As used herein, a reset element 3350 can be any device capable of switching, such that voltage source 3345 is applied to light sensitive element 3390.In one particular embodiment, reset element 3350 is a CMOS transistor that includes a reset gate 3351, a reset drain 3353, and a reset source 3352.Yet further, for purposes of this document, light sensitive element 3390 is any device that is capable of detecting the presence of light and producing a signal indicative of the amount of light detected.\",\n \"Thus, in one embodiment of the present invention, light sensitive element 3390 is a photodiode.\",\n \"In other embodiments, light sensitive element 3390 is a photo-gate.\",\n \"Other such light sensitive elements can be used including, but not limited to, a bipolar photo-transistor and/or a stacked complex n-p-n-p device where each junction is sensitive to a part of the light spectrum to be detected.\",\n \"Also for purposes of this document, selection element 3370 can be any device capable of presenting a signal representative of an amount of light impinging upon light sensitive element 3390.Thus, selection element 3370 can be a transistor, a pass gate, or the like.\",\n \"In one embodiment of the invention, selection element 3370 is a CMOS transistor with a selection gate 3371, a selection drain 3373, and a selection source 3372.For discussion purposes, each of reset element 3350, source follower transistor 3360, selection element 3370, and charge evacuation element 3380 are CMOS transistors and light sensitive element 3390 is a photodiode.\",\n \"However, based on the disclosure provided herein, one of ordinary skill in the art will recognize that the present invention and the principles included herewith are applicable to a number of other device types.\",\n \"As illustrated by pixel 3000, source follower drain 3363 and reset drain 3353 are both electrically coupled to voltage source 3345.Further, source follower gate 3361, a node of light sensitive element 3390, reset source 3352, charge evacuation source 3382, and charge evacuation drain 3383 are electrically coupled together at a node 3303.The other node of light sensitive element 3390 is electrically coupled to column ground 3355.Yet further, source follower source 3362 is electrically coupled to selection drain 3373 at a node 3313.Reset gate 3351 is driven by im_line_rst(i) 707, charge evacuation gate 3381 is driven by im_line_rst_n(i) 717, and selection gate 3371 is driven by im_line_sel(i) 727.Output signal 742 is driven by selection source 3372.In some embodiments, im_line_rst_n(i) 717 is the inverse, or complement, of im_line_rst(i) 707.Thus, when reset element 3350 is switched, thereby providing a low impedance path from reset drain 3353 to reset source 3352, charge evacuation element 3380 is not switched, thereby providing an open circuit between charge evacuation drain 3383 and charge evacuation source 3382.The opposite is also true.\",\n \"When reset element 3350 is not switched, thereby providing an open circuit between reset drain 3353 and reset source 3352, charge evacuation element 3380 is switched thereby providing a low impedance path from charge evacuation drain 3383 to charge evacuation source 3382.In some embodiments where im_line_rst_n(i) 717 is the complement of im_line_rst(i) 707, im_line rst_n(i) 717 is delayed such that a falling edge of im_line rst(i) 707 precedes a corresponding rising edge of im_line_rst_n(i) 717 by a period of time.\",\n \"In other embodiments, im_line rst_n(i) 717 works in relation to im_line rst(i) 707, but is not the complement of im_line_rst(i) 707, but rather a distinctly generated signal.\",\n \"The timing relationships of the various signals are discussed further in relation to FIG.\",\n \"9 below.\",\n \"FIG.\",\n \"8 illustrates a flow diagram 3400 of one method in accordance with the present invention for operating pixel 3000.In operation, im_line_rst(i) 707 is asserted such that a low impedance path exists between reset drain 3353 and reset source 3352 (block 3410).\",\n \"In particular embodiments, im_line_rst(i) 707 overdrives reset gate 3351 such that photodiode 3390 is charged as rapidly as possible.\",\n \"Further, im_line_rst_n(i) 717 is deasserted such that an open circuit exists between charge evacuation drain 3383 and charge evacuation source 3382 (block 3420).\",\n \"As such, a voltage corresponding to voltage source 3345, less the impedance drop across reset element 3350 is present at node 3303, thereby reverse biasing photodiode 3390.In this state, photodiode 3390 is charged to a level corresponding to the voltage at node 3303.Further, a negative charge is built in the channel of reset element 3350.Some time later, im_line_rst(i) 707 is deasserted such that an open circuit exists between reset drain 3353 and reset source 3352 (block 3430).\",\n \"Thus, additional charge from voltage source 3345 is not available to photodiode 3390.However, on the falling edge of im_line_rst(i) 707, the negative charge accumulated in the channel of reset element 3350 is dissipated through both reset drain 3353 and reset source 3352.Thus, part of the charge is discharged via node 3303 causing a voltage drop (i.e., noise), which can be significant.\",\n \"In some cases, the voltage drop due to the negative charge from reset element 3350 can be between two-hundred and five-hundred millivolts for geometries of four to seven micrometers, and is exacerbated as pixel geometries and source voltages decrease.\",\n \"With an open circuit between reset drain 3353 and reset-source 3352, the charge built up in photodiode 3390 begins to dissipate at a rate corresponding to the amount of light impinging upon photodiode 3390.Thus, where a significant amount of light impinges upon photodiode 3390, the voltage at node 3303 will decrease at a more rapid rate than if a small amount of light impinges upon photodiode 3390.Either some time after the deassertion of im_line_rst(i) 707 or coincident therewith, im_line_rst_n(i) 717 is asserted such that a low impedance path exists between charge evacuation drain 3383 and charge evacuation source 3352 (block 3440).\",\n \"This assertion provides a path whereby at least a portion of the negative charge that was stored in the channel of reset element 3350 can dissipate through charge evacuation device 3380.In some embodiments, the type of device used to implement charge evacuation element 3380 is chosen such that the operation of charge evacuation element 3380 reflects the non-linearities exhibited by reset element 3350.Thus, in some cases, charge evacuation element 3350 and reset element 3350 are chosen to be the same device types.\",\n \"As approximately half of the charge built up in the channel of reset element 3350 dissipates through reset drain 3353 and the other half dissipates through reset source 3352, the size of charge evacuation device 3380 can be chosen such that it can evacuate roughly half of the charge built up in the channel of reset element 3350.Thus, in one embodiment, where the charge evacuation element 3380 and reset element 3350 are the same device type, the size of charge evacuation device 3380 is roughly half the size of reset element 3350.In other embodiments where reset element 3350 and charge evacuation element 3380 are different device types, such as, when charge evacuation element 3380 is a switched capacitor, the size of the capacitor is chosen based on the amount of charge to be dissipated from the channel of reset element 3350 via reset source 3352.As illustrated below in FIG.\",\n \"9, dissipation of the negative charge in the channel of reset element 3350 through charge evacuation element 3380 can, depending upon the geometry of pixel 3000, eliminate as much as five-hundred millivolts of noise at node 3303.Elimination of such noise at node 3303 assures that the signal sampled at output 742 more accurately reflects the amount of light detected at photodiode 3390.At some point, im_line_sel(i) 727 is asserted to sample the remaining voltage at node 3303, and thereby gain an approximation of the light impinging upon photodiode 3390 (block 3450).\",\n \"With im_line_sel(i) 727 asserted, a low impedance path exists from selection drain 3373 to selection source 3372.Thus, a signal representative of the voltage present at node 3303 is presented on output 742 via source follower transistor 3360.Once the output is sampled, im_line_sel(i) 727 is deasserted (block 3460) and the process is repeated to detect the amount of light impinging upon photodiode 3390 at a future point in time.\",\n \"It should be recognized from FIG.\",\n \"9 that in some embodiments of the present invention, the assertion of im_line_sel(i) 727 overlaps the assertion of im_line_rst(i) 707.Yet, in other embodiments, the assertion of im_line sel(i) 727 does not overlap the assertion of im_line_rst(i) 707.FIG.\",\n \"9 illustrate timing diagrams of the various signals related to the operation of pixel 3000.More particularly, the figures illustrate the relationship between im_line_rst(i) 707, im_line rst_n(i) 717, im_line_sel(i) 727, the voltage at node 3303, and the voltage at output 742.FIG.\",\n \"9a is a timing diagram 3501 illustrating operation of pixel 3000 where im_line_rst(i) 707 and im_line_rst_n(i) 717 are complementary, and where charge evacuation element 3380 is operational.\",\n \"In contrast, FIG.\",\n \"9b is a timing diagram 3502 illustrating similar operation of pixel 3000 where charge evacuation element 3380 is not present.\",\n \"As illustrated in FIG.\",\n \"9b, at the falling edge of im_line_rst(i) 707 (noted as 3520), a voltage drop, VDROP 3510, is exhibited at node 3303.This voltage drop exists until photodiode 3390 is again charged by assertion of im_line_rst(i) 707 (noted as 3530).\",\n \"This voltage reduction at node 3303 (i.e., noise) causes a corresponding reduction in output signal 742.This is in comparison to timing diagram 3501 of FIG.\",\n \"9a, where the voltage drop at node 3303 does not occur because the charge causing the voltage drop in timing diagram 3502 is evacuated through charge evacuation element 3380 as previously described.\",\n \"Thus, as illustrated in the contrast between FIG.\",\n \"9a and FIG.\",\n \"9b, a substantial amount of noise can be eliminated from node 3303 via the operation of charge evacuation element 3380.FIG.\",\n \"9c is a timing diagram 3503 that illustrates another embodiment where, similar to timing diagram 3501, im_line_rst(i) 707 and im_line_rst_n(i) 717 are complementary.\",\n \"However, in contrast to timing diagram 3501, timing diagram 3503 illustrates im_line_rst_n(i) 717 delayed from im_line_rst(i) 707 by a period 3570.As illustrated, charge evacuation element 3380 still dissipates the charge from the channel of reset element 3350, albeit at a slightly later time corresponding to the assertion of im_line_rst_n(i) 717.Thus, a voltage droop 3580 corresponding to VDROP 3510 (illustrated as a dotted line) is noted on node 3303, until the assertion of im_line_rst_n(i) 717 (noted as 3590).\",\n \"However, because of the operation of charge evacuation element 3380, the voltage at node 3303 is corrected before the sample is taken upon assertion of im_line_sel(i) 727.Based on the disclosure provided herein, one of ordinary skill in the art will recognize a number of other timing relationships between the various signals that each dissipate the charge within the channel of reset element 3350, thus reducing the noise manifest at output 742.Further, as previously mentioned, im_line_rst(i) 707 and im_line_rst_n(i) 717 can be separately generated to achieve a timing relationship that allows for charging of photodiode 3390, and for dissipating the charge within the channel of reset element 3350.In some cases, it can minimize the logic in TCAG 136 to have im_line_rst_n(i) 717 be the complement of im_line_rst(i) 707.Further, it should be recognized that im_line_sel(i) 727 can be generated at other times than those illustrated in FIG.\",\n \"9.FIG.\",\n \"10 illustrates the architecture of column amplifier 703.Discharge (element 777) and enable (element 767) signals are received from column logic block 739.Using column amplifier 703, the output of each column is double sampled by the track and hold circuits in the column amplifier that are controlled by im_smp_strobe (element 747) and im_ref_strobe (element 757) signals.\",\n \"A timing diagram 893 illustrating the inter-relationship of the various signals of pixel array 700 and column amplifier 703 is provided as FIG.\",\n \"11.Of note, the exposure control described in more detail below should not have a line in pixel array 700 during a period 892 outlined by dotted lines on FIG.\",\n \"11.Imaging unit 130 can be programmed to provide a standard output from pixel array 700 or a mirrored image.\",\n \"FIG.\",\n \"12 provides a timing diagram of the signals associated with outputting the image in standard mode, and FIG.\",\n \"13 provides a timing diagram of the signals associated with outputting the image in mirrored mode.\",\n \"Selection between the standard and mirrored modes is accomplished by programming an im_mirror (element 787) signal via a register bit, which drives column logic block 739.Referring again to FIG.\",\n \"4, analog mux 709 selects the pixel from pixel array 700 that will be processed.\",\n \"Thus, a column of pixel array 700 is selected by column logic block 739, and then a single pixel within the column is selected via analog mux 709.Analog mux 709 is a three level mux where the first and second levels are eight to one muxes, and the third level is a six to one mux.\",\n \"This provides for a total of a 384 to one mux.\",\n \"Selection is accomplished via column address signals (element 797) im_col_add_L[7:0] for selecting the first level mux im_col_add_M[7:0] for selecting the second level mux, im_col_add H[5:0] for selecting the third level mux.\",\n \"Sensor 132 can operate in a rolling mode, in which the exposure and readout processes are linked.\",\n \"The sampling of each row data is performed at the end of the integration time.\",\n \"After sampling a selected-row data, the row is reset.\",\n \"The row is sampled again after the reset.\",\n \"This second sampling helps eliminate fixed pattern noise (FPN) by implementing a correlated double-sampling procedure.\",\n \"The integration of that row starts again according to the exposure parameters.\",\n \"A set of registers defines the total size of pixel array 700 and the size of the array that is read out and sent to translation unit 150 for further processing.\",\n \"Another set of registers defines the exposure time that is required for pixels within pixel array 700.The exposure and read timing sequences are derived from these registers.\",\n \"The processing of a single pixel of sensor 132 is referred to as a pixel cycle.\",\n \"There are four Rclk 601 cycles in a pixel-cycle that is controlled by a timing circuit.\",\n \"An Im_Pix signal from clock unit 102 qualifies the first clock edge of the pixel-cycle.\",\n \"On activation of the timing circuit, line and pixel counters are set to zero, and subsequently begin counting pixel-cycles.\",\n \"An Im_Frame 910 signal is activated on the first pixel-cycle of the frame, and an Im_Line 920 signal is activated on the first pixel-cycle of a line, except for the first line, when Im_Frame 910 is active.\",\n \"The pixel counter counts upon activation of the Im_Pix signal, and the line counter is activated when the pixel counter reaches a full-line count.\",\n \"On activation of Im_Line 920 after activation of Im_Frame 910, TCAG 136 reads and resets the first line in pixel array 700.On each activation of Im_Line 920, the lines are only reset.\",\n \"Once the line whose index is equal to the Im_starty register value is reached, data is sent out starting at the pixel whose index matches the Im_startx register value.\",\n \"The pixels that are inside the window defined by the Im_starty and Im_startx, Im_endy, and Im_endx converted from the analog to digital domain using ADC 134, and passed to translation unit 150.An additional window can be defined for differentiating between pixels in “area of interest”, and pixels outside this defined area.\",\n \"An Im_window signal is active for those pixels inside the window, and inactive for the pixels outside the window.\",\n \"Im_Line 920 starts the readout process of an image line, and also causes a reset to that line.\",\n \"One or more register values define the reset time for the first line.\",\n \"Further, a coarse exposure register value defines the number of lines between Im_Frame 910 signal activation and the end of the coarse reset time.\",\n \"Once the appropriate Im_Line 920 is reached, the number of pixel-cycles is counted according to a fine exposure register value.\",\n \"Once the reset time has elapsed, the integration time of the first line is initiated.\",\n \"The event is marked by the activation of Im_Expline 930 for one pixel-cycle.\",\n \"A second nine-bit counter is then loaded with the full-line length register and decremented on each pixel cycle.\",\n \"Once the nine-bit counter reaches a zero value, the integration time of the second line is initiated.\",\n \"Im_Expline 930 is activated for one pixel-cycle and the process repeats itself until the integration time of the last line is initiated.\",\n \"FIG.\",\n \"14 illustrates the interaction of the previously described frame timing signals.\",\n \"Each activation of Im_Expline 930 starts the exposure of an image line, and In_Line 920 signal starts the readout and reset processes.\",\n \"As illustrated in FIG.\",\n \"15, the line is sampled twice, once at the end of the exposure time (element 1010), and for the second time right after the activation of the reset signal (element 1020).\",\n \"Double buffering can be used for all frame parameters.\",\n \"This allows the Host or programmable processing core 112 to write the next frame parameters at any time.\",\n \"Implementation of the newly written parameters are, however, delayed until activation of the following Im_Frame 910 to avoid problems with the currently processing frame.\",\n \"Referring again to FIG.\",\n \"4, a signal created by a selected pixel within pixel array 700 is passed through analog mux 703 and source selection mux 709 prior to further processing by sensor 132.More particularly, the selected pixel signal is level shifted by level shift block 719, black level corrected by black offset block 723, and amplified by first stage programmable gain amplifier 729, and second stage programmable gain amplifier 733.The output of the amplified and corrected pixel signal is then passed from sensor 132 to ADC 134.In one particular embodiment of imaging device 100, the level shift, and black offset are done in the digital domain.\",\n \"In other embodiments, the processes are performed in the analog domain, while in yet other embodiments, the level shift and amplification are performed in the analog domain, while the black offset is performed in the digital domain.\",\n \"Thus, it should be recognized that the point at which a selected pixel signal is presented to ADC 134 can be changed depending upon the particular embodiment.\",\n \"Further, the order in which amplification, black offset and level shifting are performed can be modified.\",\n \"For example, the selected pixel signal can be level shifted and amplified in the analog domain, passed to ADC 134, and subsequently black level corrected by black offset block 723 in the digital domain.\",\n \"As further discussed below, analog signal processing portions of imaging device 100 are designed to ensure that as much as possible of an image detected by pixel array 700 is passed from the analog domain to the digital domain.\",\n \"Further, power and bandwidth considerations are entertained in deciding whether the processing is performed in the analog or digital domain.\",\n \"Thus, the order of processing through ADC 134, level shift block 719, black offset block 723, amplifiers 729 and 733, as well as whether the blocks are implemented in the analog or digital domain can be adjusted to ensure that the entire resolution of ADC 134 is exploited, adjusted based upon power and bandwidth considerations, and/or a combination thereof.\",\n \"As used herein, dark level is the level of the pixel signal due to leakage and other circuit related effects that occurs in unexposed pixel during a given exposure time.\",\n \"The dark level can be obtained by reading the pixels of the dark rows.\",\n \"Black level is the level of the pixel signal corresponding to the darkest pixel in a frame (e.g., shadowed areas of an image).\",\n \"White level is the level of the pixel signal corresponding to the brightest pixel in a frame (e.g., highlighted areas of an image).\",\n \"As illustrated in FIG.\",\n \"4, the selected pixel signal passed to ADC 134 is described by the equation: V adc = { ( V max - Δ V ref ) * ( 1 + im_lsh _gain ) - ( - 1 ) im_black [ 4 ] Δ V ref im_black [ 3 : 0 ] 16 } * 2 ( 1 + im_gain _m [ 2 : 0 ] ) 1 + im_gain _d [ 1 : 0 ] The signals in the analog processing chain are differential voltages in the range negative one volt to positive one volt.\",\n \"The uncorrected and un-amplified output from source selector mux 713 is in the range of zero volts to one and one-half volts.\",\n \"Level shift block 719 converts the output from source selector mux 713 to a voltage range compatible with the other analog processing circuits.\",\n \"In some embodiments, level shift block 719 is operable in one of two different modes: a high gain mode and a high dynamic range mode.\",\n \"The high gain mode is selected by setting a register output im_lsh_gain 714 equal to a high level.\",\n \"The high gain mode operates by mapping a zero to one volt input range to a negative one volt to positive one volt range.\",\n \"Alternatively, the high dynamic range mode is selected by setting a register output im_lsh_gain 714 equal to a low level.\",\n \"The high dynamic range mode operates by mapping a zero to two volt input range to the negative one volt to positive one volt range.\",\n \"FIG.\",\n \"16 illustrates the correction and amplification of the selected pixel signal as it is received from source select mux 713 and passed through level shift block 719, black offset block 723, and amplifier stages 729, 733 for both high gain mode (signal 844) and high dynamic range mode (signal 846).\",\n \"For a given exposure time, the voltages of most pixels in the frame will be in the range between the black level and the white level.\",\n \"In some embodiments, ADC 134 ideally converts only this voltage range, without losing resolution by eliminating signals outside of the range.\",\n \"In one particular embodiment, adjusting the voltages of the pixels to be in the range between black level and white level is achieved by black offset block 723.To this end, a control signal, im_black[4:0] (element 724), can be provided in sign-magnitude format and use to program the level adjustment to the proper level.\",\n \"In one embodiment, the resolution of black offset block 723 is equivalent to the thirty-two least significant bits of ADC 134.FIG.\",\n \"17 illustrates examples of signal outputs for normally lighted (signal 853), dark (signal 857), and bright (signal 859) subjects from source select mux 713 that are passed through level shift block 719, black offset block 723, and amplifier stages 729, 733.The gain and black level values are all pixel color dependent.\",\n \"The red-green and blue-green ratios of the gain settings are determined in order to achieve gross white balance.\",\n \"An exemplary timing of the control signals for the analog processing circuits is presented in FIG.\",\n \"18.A sel[1:0] (element 774) is representative of the timing of column address signals 797 and also and im_test[7:0] (element 704) signal.\",\n \"In one particular embodiment, a maximum exposure time is one frame at thirty frames per second, or 33.33 milliseconds, and a maximum gain of the analog chain is thirty-two (i.e, two from level shift block 719, and four from each of amplifiers 729 and 733).\",\n \"A pixel sensitivity of 0.226 [V/lux*sec] for a green pixel at 540 nm is provided by pixels within pixel array 700.Full scale of ADC 134 is two volts (e.g., negative one volt to positive one volt differential), where the least significant bit is 1.95 mV.\",\n \"Using these parameters, the size of the analog processing section of imaging device 100 can be determined and optimized.\",\n \"For determining the resolution used to obtain the proper exposure, the lowest illumination level that can be translated to full scale response of ADC 134 corresponds to the maximal exposure time and maximal amplifier gain.\",\n \"The following equation describes the approach: 2[V]/(32*0.0333[sec]*0.226[V/lux*sec])=8.3[lux] If the highest level of illumination is 100,000 [lux], the lowest exposure time needed assuming minimal amplifier gain of unity is: 2[V]/(1*100,000[lux]*0.226[V/lux*sec])=88.5 [usec] As in a CIF frame there are 101,376 pixels, the minimal exposure time corresponds to about three-hundred, fifty-four pixel cycles at thirty frames per second.\",\n \"Therefore, controlling exposure time at the resolution of one pixel cycle is adequate.\",\n \"For sizing black offset block 723, it is assumed that the exposure is set so that at the output source select mux 713, the white level corresponds to a drop of one volt and the black level corresponds to a drop of 937.5 mV.\",\n \"If the amplifier 729, 733 gain is set to sixteen, and the black level DAC is five-bit (sign magnitude) with the least significant bit equaling 62.5 mV is set to fifteen, this will stretch the narrow histogram on the full resolution of ADC 134.On the opposite end, for a dark picture where at the output of source select mux 713 the black level is zero and the white level corresponds to a drop of 62.5 mV, a setting of negative fifteen for the black level DAC and a gain of sixteen for the amplifier 729, 733 will stretch the narrow histogram on the full resolution of ADC 134.The programmable gain amplifier 729, 733 has a maximum gain of sixteen and supports several functions including extending the exposure range for low illumination levels, providing gross white balance, and stretching the histogram as described in relation to black level correction.\",\n \"Programmable gain amplifier 729, 733 can be implemented as a two stage amplifier, where each stage exhibits a maximum gain of four.\",\n \"First stage programmable gain amplifier 729 is described by the following equation: Gain PGA_D = 4 1 + im_gain _d [ 1 : 0 ] Second stage programmable gain amplifier 733 is described by the following equation: Gain PGA_M = 1 + im_gain _m [ 2 : 0 ] 2 There is separate gain control for each color filtered pixel.\",\n \"The parameters associated with the gain control are used for coarse white balance, and for enhanced exposure in dim light situations, where the maximum exposure time has been reached, but the output image is still too dark.\",\n \"The gain parameters are used for expanding the range of the sampled data to the full range of ADC 134.The first sixteen lines of pixel array 700 are dark lines that are not exposed to light.\",\n \"Each of the lines are integrated similar to other lines and the dark level for each color is separately measured on the output of ADC 134.The average of the measured dark line values is passed back for Dark pixel correction of the lines that follow the dark lines in pixel array 700.The effect can be immediate, where the calculation result is activated during the readout of the same frame where the dark lines were evaluated.\",\n \"Image device 100 includes two methods for calculating the dark offset.\",\n \"One or the other mode is selected by writing a predetermined mode selection in a register.\",\n \"The two modes are dark offset per column and average dark offset.\",\n \"Dark offset per column involves calculating and storing an offset value for each column.\",\n \"When reading the image area, the dark offset parameter selected based on the particular column that is currently being read from pixel array 700, where the line number indicates the color being read.\",\n \"The offset for each column is averaged on the fifth to twelfth lines, giving a total of eight lines.\",\n \"The offset for each color component in each column is c averages of four pixels of the same column and color, minus the fixed black offset that is applied during these lines.\",\n \"During the dark offset calculation, the black offset inputs are set to a positive known value and stored in a register, the color dependent gain is applied, and the output of ADC 134 is sampled.\",\n \"Two RAMs of 384 by twelve-bits can be used.\",\n \"The output from ADC 134 for the first and second dark lines is written to the RAMs.\",\n \"For the following lines, the even-numbered line values are added to the first RAM entries, and the odd-numbered line values to the second RAM entries.\",\n \"After the seventh line (a “RED” line), values are added and the result is stored back in the RAM.\",\n \"The next line is processed in the same manner for the “BLUE” line offset.\",\n \"Throughout this process, the average offset per color is calculated as well.\",\n \"During the period for lines twelve and thirteen, which are dark lines that are not read from the array, the RAM contents are read, and the “column noise” is calculated as follows: Column—offset=(RAMsum)/4, rounded up Noise=Column—offset−Average, 12 bits two's complement.\",\n \"When reading the image pixels from pixel array 700, the “noise” dark offset is read from the RAM.\",\n \"The correction is performed in the digital domain on the output of ADC 134.The values that are stored in the RAM are subtracted from the output of ADC 134 according to the column number and the line type that is being read.\",\n \"To simplify calculations, average dark offset mode is utilized.\",\n \"In the average dark offset mode, the first four lines are skipped, and only the next eight dark lines are evaluated.\",\n \"Evaluation is performed from pixel number sixty-four to pixel number three hundred nineteen, totaling 256 pixels per line.\",\n \"A total of 128×2 green, 128 blue and 128 red pixel values are collected.\",\n \"The result is averaged, limited, and stored in registers.\",\n \"Further, in some embodiments, three four-bit black offset parameters are stored in a register for each of the three colors.\",\n \"The value in the registers are passed to imaging unit 130 and subtracted from or added to the analog image signal before the signal is passed to ADC 134.Thus, three four-bit digital to analog converters (“DAC”) are used to convert the register values to the analog domain before the subtraction process is accomplished.\",\n \"The operation is performed in accordance with a sign bit that is passed along with each of the three register values.\",\n \"In some instances, the four-bit offset values are each multiplied by sixteen before the subtraction process is completed.\",\n \"In addition, some embodiments of imaging device 100 include provisions for reducing random noise.\",\n \"Such provisions include, but are not limited to, applying the aforementioned charge injection technique to pixel reset signals, stopping operation of the various digital circuitry in imaging device 100 while each row of pixel array 700 is sampled, and providing a clock driving the analog circuitry that precedes the clock driving the digital circuitry to keep various substrate noise effects orthogonal in time domain.\",\n \"In one embodiment, such a phase difference between the analog clock and the digital clock is provided by deriving the analog clock from the falling edge of the digital clock, and subsequently adding some delay to the analog clock before applying it to ADC 134.Some embodiments include additional measures to eliminate random noise that include, but are not limited to, providing analog processing switched capacitor circuits that use clock phases generated by ADC 134 and synchronizing control signals driving the analog circuitry using the clock generated by ADC 134.In addition, the sizes of the capacitors used in the switched capacitor circuits can be increased in order to meet the precision requirements and to reduced kT/Cnoise.\",\n \"Some embodiments of the present invention provide for reducing power consumption of imaging device 100.Such processes can include reducing power consumption of the analog processing circuitry of imaging device 100, while in other embodiments, power consumption is reduced by reducing power consumption of the digital processing circuitry of imaging device 100.In one particular embodiment, power consumption is reduced by limiting power consumption by both the analog processing circuitry and the digital processing circuitry of imaging device 100.FIG.\",\n \"19 is a timing diagram illustrating the relationship between power consumption control signals, Im_stdby 764 and im_adc_cnt[4] 754.As previously illustrated in FIG.\",\n \"4, Im_stdby 764 is provided to the various analog processing circuits of sensor 132 including analog mux 709, source selection mux 713, level shift block 719, black offset block 723, and programmable gain amplifier 729, 733.Im_adc_cnt[4] 754 is provided to ADC 134.Following FIG.\",\n \"19, when Im_stdby 764 is asserted high (noted 1920), the analog processing circuitry stops functioning during a non-functional period 1910 surrounded by dashed lines.\",\n \"One cycle of aclk 603 after assertion of Im_stdby 764, Im_adc_cnt[4] 754 is asserted high (noted 1940), thus placing ADC 134 in the standby mode.\",\n \"To awake from the standby mode, Im_adc_cnt[4] 754 is deasserted (noted 1950) and twenty cycles of aclk 603 later (noted 1960), Im_stdby 764 is deasserted (noted 1970).\",\n \"Once Im_stdby 764 is deasserted, the analog processing circuitry becomes operational again one cycle of aclk 603 later (noted 1980).\",\n \"During non-functional period 1910, the analog processing circuits will enter a low power consumption standby mode.\",\n \"In one embodiment of the standby mode, the common mode feedback of the operational amplifiers remains operational, but the circuits associated with the various analog processing calculations are disabled.\",\n \"This maintains the inertial state of the amplifiers, while reducing the power consumption of the analog circuitry associated with imaging device 100.FIG.\",\n \"20 illustrates a block diagram 1905 of circuitry related to controlling the standby mode.\",\n \"Block diagram 1905 includes pixel array 700 providing differential inputs 1903 to a processing control system 1925.Processing control system 1925 provides either differential inputs 1903 to an analog processing circuit 1955 via differential inputs 1906, or a differential reference voltage 1915 (received via inputs 1904).\",\n \"Differential inputs 1906 are processed by analog processing circuitry 1955 as previously discussed, and the results presented via differential inputs 1907 to ADC 134 are converted to the digital domain and passed to a digital processing circuit 1975 via a bus 1965.Digital processing circuit 1975 performs the various digital processing functions as previously discussed, and the processed data is provided as an image output.\",\n \"In some embodiments analog processing circuit 1955 includes column amplifier 703, analog mux 709, source selection mux 713, level shift block 719, black offset block 723, and programmable gain amplifier 729, 733.In other embodiments, analog processing circuit 1955 includes a subset of the aforementioned element.\",\n \"For example, in one particular embodiment, analog processing circuit 1955 includes level shift block 719, black offset block 723, and programmable gain amplifier 729, 733.Thus, depending upon the design, one or more of the aforementioned blocks can be placed in standby mode either individually, or as a group.\",\n \"Further, in some embodiments, digital processing circuit 1955 includes translation unit 150 and parallel interface unit 190.In other embodiments, digital processing circuit 1955 includes only one or the other of translation unit 150 and parallel interface unit 190.In some cases, only a subset of the pixels within pixel array 700 are processed and provided as an image output.\",\n \"For example, bandwidth available for the image output may be less than that required to output each pixel of pixel array 700.As a more specific example, every other pixel line and/or pixel column of pixel array 700 may be dropped.\",\n \"In such cases, embodiments of the present invention provide for limiting power consumption of imaging device 100 by placing analog processing circuit and/or digital processing circuit 1975 in the standby mode.\",\n \"To do this, a sub-sample control system 1945 provides an indication about whether a particular line, column and/or pixel is to be processed.\",\n \"This indication From sub-sample control system 1945 is used to generate Im_stdby signal 764 which, as previously discussed, controls whether analog processing circuit 1925 and/or digital processing circuit 1975 are to enter the standby mode, or remain operational.\",\n \"In some embodiments, sub-sample control system 1945 is a hardware system utilizing information from frame rate block 114 of control unit 110 and/or TCAG 136 of imaging unit 130 to determine where in a frame a received pixel is from, and whether the pixel is to be processed.\",\n \"Alternatively, in other embodiments, sub-sample control system 1945 is implemented in software and executed by programmable processing core 112.In yet another embodiment, sub-sample control system 1945 is a hybrid hardware and software implementation utilizing relevant portions of TCAG 136, frame rate block 114, and/or programmable processing core 112.In one particular example where only two of every four pixel lines are processed, sub-sample control system 1945 monitors the addressing of pixel array 700 and causes Im_stdby 764 to assert each time a non-processed pixel line is addressed.\",\n \"This causes processing control system 1925 to present reference voltage 1915 to analog processing circuit 1955 in place of differential input 1903 from pixel array 700.Reference voltage 1915 is electrically coupled to switching inputs of analog processing circuit 1955, thus reducing power dissipation due to switching circuits in analog processing circuit 1955.However, the common mode feedback of the operational amplifiers within analog processing circuit 1955 continue operating.\",\n \"This maintains the inertial state of the amplifiers, while reducing the power consumption of the analog circuitry associated with imaging device 100.In addition, Im_adc_cnt[4] 754 is asserted to place ADC 134 in a similar standby mode.\",\n \"Yet further, a digital disable signal 1985 is provided from processing control system 1925 to digital processing circuit 1975.This signal is used to gate the various clocks distributed to the various operating units of imaging device 100.In some embodiments, processing control system 1925 comprises portions of Rclk unit 102.In such embodiments, the various clock signals are gated by Rclk unit 102 and there is no reason to provide digital disable signal 1985 to digital processing circuit 1975.FIG.\",\n \"21a illustrates all exemplary circuit 1943 used to electrically couple analog circuits within analog processing circuit 1955 to reference voltage 1904.In circuit 1943, PH1 and PH2 are non-overlapping clock phases.\",\n \"In standby mode, there is no switching to vref 1904a and vref_n 1904b, rather inputs 1903 are shorted to vcommon 1953 to limit switching.\",\n \"With such a circuit, both the bias currents of the Op-Amps, as well as the switching related to analog calculations is reduced.\",\n \"However, switching sufficient to keep the common mode feedback loops of an amplifier 1963 active is maintained.\",\n \"When Im_stdby 764 is asserted high, standby switches 1964 are shut and operational switches 1965 are open.\",\n \"Thus, switching in circuit 1943 is limited.\",\n \"FIG.\",\n \"21b illustrates exemplary circuit 1943 when Im_stdby 764 is deasserted, and circuit 1943 is operating in phase one, PH1.FIG.\",\n \"21c illustrates exemplary circuit 1943 when Im_stdby 764 is similarly deasserted, but circuit 1943 is operating in phase 2, PH2.FIG.\",\n \"22 is a flow diagram 1974 illustrating a method according to the present invention for entering and exiting the standby mode.\",\n \"Following flow diagram 1974, parameters associated with sub-sample control system 1945 are set-up (block 1976).\",\n \"This can include writing various registers controlling the portions of pixel array 700 that are processed.\",\n \"Such a register write can be performed by programmable processor control 112 and/or the Host.\",\n \"An image is detected from pixel array 700 (block 1977).\",\n \"Pixel array 700 is addressed by TCAG 136 to cause a desired column of pixel signals to be presented by pixel array 700 (block 1978).\",\n \"If sub-sample control system 1945 indicates that the column is to be processed (block 1979) and that the line is to be processed (block 1981), then the analog processing circuitry 1955 is enabled, or awakened from the standby mode (block 1982).\",\n \"Further, digital processing circuitry 1975 is enabled by removing the clock gate (block 1983).\",\n \"With the circuitry thus enabled, the pixel signals received from pixel array 700 are processed.\",\n \"Alternatively, where either the pixel column (block 1979) or the pixel line (block 1981) is not to be processed, analog processing circuit 1955 (block 1986) and digital processing circuit (block 1987) are placed in the standby mode to conserve power.\",\n \"In such a state, the pixel signals received from pixel array 700 are dropped and not processed (block 1988).\",\n \"The power distribution and management scheme for the analog circuitry provides a supply of 2.8 volts +/−10% supply.\",\n \"On-chip regulators are used to provide the following supply voltages: —2.5 volts +/−10% for the 0.25 u digital circuits, 2.4 volts +10/−5% for the imager array on a vddArr 881 supply, and 3.2 volts +10/−5% for the imager array line reset drivers.\",\n \"A block diagram 896 of the power management for imaging device 100 is illustrated in FIG.\",\n \"23.As illustrated, the regulators are controlled by the following signals: a reg_d enable 882 signal the when asserted low causes the digital regulator to enter standby mode with the 2.5 V voltage is supplied from an external source.\",\n \"In order to enable power up, reg_d_enable 882 is gated with an atest 883 signal as shown in FIG.\",\n \"23.At power up atest 883 is kept low forcing reg_d_enable 882 high.\",\n \"When reg_d_enable is 882 is high, the digital regulator output voltage is reduced to 2.0 V. The order of power-up of the regulators has to be digital regulator 884 first and only then a boost regulator 885.The clock of boost regulator 885 has to be provided by the digital circuits.\",\n \"It is a clock in the range of 500 KHz to 1 MHz, obtained by dividing Rclk 601 by sixteen.\",\n \"The edges of the divided clock should be aligned with the digital signals (i.e., aligned with the rising edge of Rclk 601).\",\n \"To facilitate testing and/or characterization, imaging device 100 includes one or more built in self test (“BIST”) routines.\",\n \"In particular embodiments, the BIST routines provide the ability to test one or more of blocks 700, 703, 709, 713, 719, 723, 729, 733, and/or 134 using a digital tester.\",\n \"In some cases, such testing can further be accomplished with or without illumination of imaging device 100.Such approaches can reduce the costs and increase the rate at which the functionality of imaging device 100 is verified.\",\n \"In some embodiments, one or more additional lines of “pseudo”-pixels are included within pixel array 700.The output from the pseudo-pixels can be a pre-defined voltage that remains constant regardless of any light impinging upon pixel array 700.For example, in one instance, the pseudo-pixels have an output of one volt that is passed to the column amplifiers.\",\n \"These pixels are activated by asserting im_line_test (element 737).\",\n \"The outputs of the pseudo-pixels are passed through column amplifier 703, and analog mux 709.In turn, the outputs are passed though source selector mux 713 by proper setting of im_test[8:0] (element 704).\",\n \"Im_test[8:0] (element 704) is derived from decoding a programmable digital register (im_test_en[3:0]).\",\n \"The outputs are then passed through one or more of blocks 719, 723, 729, 733, and 134.The output of ADC 134 can then be compared against a range of values to determine if imaging device 100 is within acceptable design limits.\",\n \"The comparison can be performed either by a programmable hardware comparator, a hardwired hardware comparator, or via programmable processor core 112.Such a test provides an ability to test the operation of column amplifier 703, and/or one or more of blocks 719, 723, 729, 733, and 134.As the inputs and outputs associated with the test are available in the digital domain, a purely digital test of the analog circuit can be performed.\",\n \"This provides an efficient and cost effective means for verifying the functionality of imaging device 100 as testing of the analog circuitry can be performed on a digital tester.\",\n \"As previously stated, an output to be tested using one or more of the BIST routines can be passed through one or more of blocks 719, 723, 729, 733, and 134.Which blocks the outputs pass through is programmable.\",\n \"Using such a programmable approach, each block can be individually bypassed.\",\n \"Such bypassing allows for the output of each block individually, or a combination of the blocks to be tested to be isolated for testing purposes.\",\n \"Bypassing of the individual blocks is controlled by signals im_by_lsh 702 to bypass level shift block 719, im_by_blk 712 to bypass black offset block 723, im_by_pgd 722 to bypass first stage programmable gain amplifier 729, and im_by_pgm 732 to bypass second stage programmable gain amplifier 733.Further, in some embodiments, source selector mux 713 permits application of various test voltages to the input of the analog processing circuitry.\",\n \"This provides for applying known analog voltages to one or more portions of analog processing blocks 719, 723, 729, 733, 134.In one embodiment, the digital output generated by applying the known analog voltage to the blocks is compared against a predetermined value, or range of values.\",\n \"In this way, testing of analog blocks 719, 723, 729, 733, 134 can be performed as a purely digital test, where im_test (8:0) (element 704) is generated from a programmable digital register (im_test_en[3:0]), and the outputs are each tested in the digital domain.\",\n \"Again, this provides an efficient and cost effective means for verifying the functionality of imaging device 100 as analog testing can be performed on a digital tester.\",\n \"Further, as each of blocks 719, 723, 729, 733 can be individually bypassed, any combination of blocks 719, 723, 729, 733 can be verified using the digital test.\",\n \"In one particular embodiment of the present invention, source selector mux 713 selects between five input voltages based on im_test(8:0) (element 704).\",\n \"More particularly, source selector mux 713 permits connecting reference voltages (v_ref 706, common 716, and vref_n 726) that are generated internal to imaging device 100 to the input of the analog processing blocks.\",\n \"In some embodiments vref 706 and vref_n 726 are derived from ADC 134, and common 716 is a provided via a resistive divider coupled to the source voltage.\",\n \"In addition, source selector mux 713 also provides for selecting externally generated reference voltages, atest 736 and atest_n 746 that can be bonded to input pads of imaging device 100.By selecting one of the five reference voltages 706, 716, 726, 736, 746, a known voltage can be applied to one or more of blocks 719, 723, 729, 733, and output via ADC 134.The output of ADC 134 can then be compared against a range of values to determine if imaging device 100 is within acceptable design limits.\",\n \"In some embodiments, an additional test mechanism provides for digital testing of each column within pixel array 700.In such a test, pixel outputs from pixel array 700 are paired with an output from reset level 743.Reset level 743 consists of a group of pixels kept under a reset state and a column amplifier identical to column amplifier 703.BIST pixels within pixel array 700 are selected and the outputs from the pixels are passed through a selected portion of column amplifier 703 and analog mux 709.One side of the output is then paired with a known reset level 743 signal and selected via source selector mux 713.This pairing between the known reset level 743 and the output from the various BIST pixels is then passed through one or more of blocks 719, 723, 729, 733, and 134.The output of ADC 134 can then be compared against a range of values to determine if imaging device 100 is within acceptable design limits.\",\n \"Such a test provides an ability to test the operability of each column amplifier within column amplifier 703, using a digital tester and without requiring illumination of imaging device 100.The various BIST modes for one particular embodiment are described in Table 4 below.\",\n \"TABLE 4 Test Modes For Analog Blocks Negative Positive output of output of source source selector im_test selector mux 713 mux 713 im_test_en[3:0] active connected to connected to Comments 0000 0 Positive output of Negative output Normal analog mux 709 of analog mux 709 operation with CDS 0001 1 Reset level 743 Negative output Normal of analog mux operation 709 without CDS 0010 2 Reset level 743 Positive output Testing reset of analog mux 709 sampling branch of column amplifier 0011 3 atest 736 atest_n 746 Testing with external signal 0100 4 vref_n 726 vref 706 −1 V 0101 5 common 716 vref 706 −0.5 V 0110 6 common 716 common 716 0 V 0111 7 common 716 vref_n 726 0.5 V 1000 8 vref 706 vref_n 726 1 V In one particular embodiment ADC 134 is a ten-bit ADC with pipeline architecture.\",\n \"The ten-bit output from ADC 134 is passed to translation unit 150.For the various dark-current calculations, four ten-bit outputs are averaged, and the most significant ten-bits of the average value are used.\",\n \"Translation Unit Translation unit 150 provides color processing for imaging device 100.Parameter and mode registers associated with translation unit 150 are loaded via control bus 101.A Bayer image is received from imaging unit 130, processed by translation unit 150, and transferred to parallel interface 190 for output via parallel interface I/O 192.While processing the image, statistics are calculated by translation unit 150.After completing the processing of an image, translation unit 150 produces an interrupt to control unit 110 indicating that processing is complete and that the statistics are ready to be read by control unit 110 via control bus 101.Translation unit 150 receives the Bayer grid as input from imaging unit 130.For each received pixel, two color values are missing and need to be reconstructed.\",\n \"Translation unit 150 reconstructs the RGB set for each pixel, performs various correction algorithms, and generates a YUV representation of the image.\",\n \"FIG.\",\n \"24 provides a block diagram 1100 illustrating translation unit 150 in greater detail.\",\n \"As illustrated, translation unit 150 includes an RGB reconstruction block 1110 (for demosaicing), a CIF-to-QCIF reduction block 1130, a color and white-balance correction block 1140, a gamma look-up table (“LUT”) 1150, an RGB to YUV generation block 1160, a twenty-four bit YUV to 4:2:2 conversion block 1170, and a statistics block 1195.Statistics block 1195 includes a white balance statistics block 1150, an exposure statistics block 1185, and a G line-sum block 1190 for providing a sum of G Bayer components per line used to detect flicker.\",\n \"RGB reconstruction block 1110 receives the Bayer input from imaging unit 130 and calculates all three color components for each pixel.\",\n \"Median Interpolation method is selected for the Bayer-to-RGB conversion.\",\n \"For green reconstruction from a pixel whose Bayer component is either red or blue, the green color component is given by the median value of the four surrounding Green Bayer components.\",\n \"The median function is defined as the average between the two values that remain when a series of four values is evaluated, and the minimum and maximum values are removed.\",\n \"For Blue reconstruction from a pixel whose Bayer component is either Red or Green, the Blue color component is given by the median value of the four surrounding Blue Bayer components.\",\n \"For a pixel whose Bayer component is green located at a blue line, the blue value is the average of the two adjacent blue Bayer components in the same line.\",\n \"Further, for a pixel whose Bayer component is green located at a red line, the blue value is the average of the two adjacent blue Bayer components in the same column.\",\n \"For red reconstruction at a pixel whose Bayer component is blue, the red color component is given by the median value of the four surrounding red Bayer components.\",\n \"At a pixel whose Bayer component is green located at a red line, the red value is the average of the two adjacent red Bayer components in the same line.\",\n \"At a pixel whose Bayer component is green located at a blue line, the red value is the average of the two adjacent red Bayer components in the same column.\",\n \"After reconstruction, the resulting RGB image is smaller than the Bayer input image by two columns horizontally, and two lines vertically.\",\n \"In some embodiments, imaging device 100 provides three methods for producing a QCIF size image.\",\n \"In the first method, a QCIF sized window is read from a CIF sized array.\",\n \"For the method, no special processing is needed and the RGB per pixel is calculated in the same manner as for a CIF sized image.\",\n \"The method provides a limited Field of View (“FOV”), but provides a good resolution.\",\n \"In the second method, the array read out of pixel array 700 is down-sampled.\",\n \"This method is implemented in the imaging unit 130.The third method is a Median Billinear method.\",\n \"In the method, the CIF sized Bayer image is first converted to an RGB image using the Median method.\",\n \"Then, if down-sampling is selected, the CIF image is reduced to a QCIF size image using a ( 1/4, 1/2, 1/4) filter—horizontally and then vertically.\",\n \"In some embodiments of imaging device 100, color correction block 1140 can be used to compensate for any disparity between the RGB filters on imaging device 100 and a display connected via parallel interface I/O 192.In some instances, sensor 132 is not equally sensitive to each of the color components.\",\n \"To compensate for the different sensitivity, and to emulate the human eye adjustment to different lighting conditions, the captured image undergoes a white-balance correction.\",\n \"The coefficients of the color correction are combined into a single matrix, and the corrected R′G′B′ values per pixel are calculated according to the following equations: R′=CR1*R+CR2*G+CR3*B+Roffset G′=CG1*R+CG2*G+CG3*B+Goffset B′=CB1*R+CB2*G+CB3*B+Boffset In addition, Gamma Correction LUT 1150 is used to gamma correct the various pixels.\",\n \"In one embodiment, Gamma correction LUT 1150 includes 512 entries, where each entry is a nine-bit value.\",\n \"The data for Gamma correction LUT 1150 is loaded to RAM within imaging device 100.Gamma correction LUT 1150 receives a color component value from color correction block 1140, and uses the value as an address to access the LUT stored in RAM.\",\n \"The value obtained from the LUT at the addressed location is stored until all three color components are received.\",\n \"Then, the three color components are passed together to RGB-to-YUV block 1160.In YUV to 4:2:2 conversion block 1170, an input of an array of pixels, each with its own R, G and B values is reconstructed into an image of YUV 4:2:2 values.\",\n \"The result is then passed to parallel interface 190.The process involves an interim conversion to a Y-Cr-Cb (4:4:4) format.\",\n \"The Y-Cr-Cb (4:4:4) is calculated as follows: R′G′B′ are given at an 8-bit accuracy, in the range of 0-255, Y is limited to the range of 16-235, and Cr and Cb are limited to the range of 16-240.Then, the following equations are used to complete the conversion: Y=0.257R′+0.504G′+0.098B′+16 Cb=−0.148R′−0.291G′+0.439B′+128 Cr=0.439R′−0.368G′−0.071B′+128 The coefficients are loaded in {s,0.8} format, eight-bits of a fraction value, without integer bits, but with a single sign bit.\",\n \"In the same manner, YUV can be calculated, without the clamping.\",\n \"The multiply coefficients are programmable.\",\n \"The offset for the luminance can be enabled or disabled.\",\n \"When offset is enabled, the value of sixteen is used as offset for the luminance.\",\n \"The chrominance offset is fixed at one-hundred, twenty eight.\",\n \"The twenty-four bit YUV value per each pixel (Y-Cr-Cb 4:4:4), is down-sampled to a YUV 4:2:2 format.\",\n \"To do this, the Y value for each pixel is retained, but each pair of consecutive luminance values (Y2n, and Y2n+1, n=0,1,2 .\",\n \".\",\n \". )\",\n \"share a pair of Cb and Cr values.\",\n \"The placement of the Chrominance pair is on the even numbered pixel.\",\n \"FIG.\",\n \"25 illustrates this down-sampling process.\",\n \"The color components of the odd number pixels are zeroed out, while the color components of the even numbered pixels are calculated as follows: For first pixel in line: Cx′[0]=Cx [0] (x is either r or b) For subsequent pixels in the line: For n=1 to n, n is equal to (N/2−1), N is the line length tmp[2n]=(Cx[2n−1])+2*(Cx[2n])+(Cx[2n+1]) (x is either r or b) The result is rounded up and divided by 4: Cx′[2n]= 1/4(tmp[2n]+2) (x is either r or b) Table 14 provides a summary of calculations performed by translation unit 150 as previously discussed.\",\n \"TABLE 13 Calculation Summary Var Formula Accur.\",\n \"Remarks Input from C C are the Bayer grid outputs 10 bits unsigned imaging unit 130 Faulty C1 C1 = C if not faulty, 10 bits unsigned correction replacement value if faulty RGB per pel C2 C2 = (C1 + C1 + 1)/2 10 bits Average and round up C10 C10 = C2 or C1 10 bits unsigned Color C20 Color correction coefficient 8 bits + sign 8 bits value, 1 bit sign correction C21 C21 = C20 * C10 19 bits 18 bits magnitude, 1 bit sign [1, 18], C22 C22 = C21r + C21b + C21g 21 bits 21 bits 2's comp [1, 20] Toff Truncation parameter 4 bits Values 0 to 10 C23 C23 = truncated C22 21 bits Programmable C23 = {sign-ext, C22[20:t]} truncation [t − 1:0] lsbits truncated, Coff Offset for Color correction 8 bits + sign 8 bits value, 1 bit sign C24 C24 = (C23 + Coff * 2 + 1)/2 21 bits offset, round up (add 1) ignore lsbit, limit.\",\n \"Lower limit 0 or 16 Loadable Upper limit (0, 511 for color correction) C25 C25 = limited C24 9 bits select bits [10:1] Gamma C30 C30 = LUT[C25] 9 bits correction RGB to Y20 Format conversion 8 bits + sign 8 bits value, 1 bit sign YUV coefficient Y21 Y21 = Y20 * C30 19 bits 18 bits magnitude, 1 bit sign [1, 18], Y22 Y22 = Y21r + Y21b + Y21g 21 bits 21 bits 2's comp [1, 20] Toff Truncation parameter 4 bits Values 0 to 10 (fixed to 8) Y23 Y23 = truncated Y22 21 bits Programmable Y23 = {sign-ext, truncation Y22[20:8]} 8 lsbits truncated Yoff Offset for conversion 8 bits + sign 8 bits value, 1 bit sign Y24 Y24 = (Y23 + Yoff * 2 + 1)/2 21 bits offset, round up (add 1) ignore lsbit, limit.\",\n \"Lower limit 0 or 16 Loadable Upper limit 235/255 for Y 240/255 for U/V Y25 Y25 = limited Y24 8 bits select bits [9:1] 4:4:4 Yout Yout = Y25 8 bits to Ctmp Ctmp = (Yn − 1 + Yn * 2 + Yn + 1) 10 bits 4:2:2 Cout Cout = (Ctmp[9:1] + 1)/2 8 bits In addition, translation unit 150 collects a number of statistics during the previously discussed translations and calculations.\",\n \"At the end of processing of a frame data, the resulting statistics are checked, and the different registers that control the correction algorithms are updated according to the results.\",\n \"The different statistics checked by the various statistics blocks including white balance block 1180, exposure statistics block 1185, and G-line-sum block 1190.The various statistics checked can include, but are not limited to, white balance, exposure, minimum and maximum per color, and flicker.\",\n \"Exposure statistics are determined by evaluating green Bayer pixels or Y output data from translation unit 150.The evaluation results in an eight-bit histogram.\",\n \"There are eight seventeen-bit registers, each holding the number of pixels whose Y or Bayer value falls within the range that is defined for that bin (ten-bit Bayer components are divided by 4 before sorting).\",\n \"At the end of a frame processing, programmable processing core 112 reads the histogram results and calculates the exposure, offset and gain parameters for the following frame.\",\n \"Table 5 lists the eight bins into which the values are assigned to create the eight bit histogram.\",\n \"TABLE 5 Exposure Histogram Bins Bin Number Range Bin #0 0-31 Bin #1 32-63 Bin #2 64-95 Bin #3 96-127 Bin #4 128-159 Bin #5 160-191 Bin #6 192-223 Bin #7 224-239 In addition, the statistics blocks of translation unit 150 detect the minimum and maximum value per each color, on the output of the Bayer-to-RGB conversion block 1110.The results can be read from registers by programmable processing core 112 at the end of the frame processing.\",\n \"Further, flicker detection can be performed using the statistics blocks of translation unit 150.Flickering can be caused by a mismatch between the frequency of light impinging upon a detected image, and the frequency at which the detected image is sampled.\",\n \"The effect of flickering is a modulation of the image information along the rows of pixel array 700.To eliminate flickering, the frequency at which the detected image is sample can be adjusted to coincide with the frequency of the light impinging upon the image.\",\n \"In one embodiment of the present invention, an estimate of the impinging light is calculated, and the sample frequency is adjusted automatically.\",\n \"In some embodiments, to obtain an estimation of the impinging frequency with high confidence, a uniform image is captured, thus avoiding detection of modulation in the image itself.\",\n \"However, in other embodiments, any image is captured and flickering is automatically detected and corrected.\",\n \"FIG.\",\n \"26 is a flow diagram of such a flicker correction system 2205 in accordance with embodiments of the present invention.\",\n \"Flicker correction system 2205 includes a feedback loop for automatically adjusting exposure to reduce flicker.\",\n \"Flicker correction system 2205 includes capturing an image (block 2206) and outputting the captured image.\",\n \"As is evident from the preceding discussion, the image is captured by sensor 132 and provided to translation unit 150 via ADC 134.The captured image received from ADC 134 is processed to determine if flickering is evident (block 2207).\",\n \"Then, the exposure is adjusted to account for any detected flickering (block 2208).\",\n \"Using the adjusted exposure parameters, the image is again captured and the adjustment process performed again.\",\n \"In some embodiments, this process can be completed for one or more frames of the image.\",\n \"This use of additional image information provides for more accurate detection and correction.\",\n \"Further, such an iterative approach allows for continuous monitoring of flicker conditions.\",\n \"In other embodiments, however, flicker detection and correction is only infrequently performed, rather than continuously.\",\n \"In addition, in some embodiments, the process may adjust exposure duration in order to emphasize flicker artifacts and thereby increase the probability of correct detection and/or reduce estimation time.\",\n \"Such an approach can include a manual adjustment whereby a user adjusts the capture frequency until the user detects a noticeable flicker.\",\n \"After that, the flicker is automatically detected and corrected.\",\n \"Alternatively, the adjustment creating the noticeable flicker can be automatically performed, and subsequently corrected.\",\n \"Alternatively, a fully passive mode of operation is also possible.\",\n \"In such a fully passive mode, exposure time is not altered by flicker detection mechanism until the estimation is completed.\",\n \"Further, such adjustments to the exposure time can be limited to multiples of a detected flicker period.\",\n \"In passive mode of operation the exposure adjustments can be introduced by other parts of the system including, for example, exposure selection sections that provide enough information for successful estimation of flicker frequency A light source, I(t), illuminating an image can have a flickering component at a frequency f that can be modeled the following way: I(t)=a·cos(2πft+φ)+b, where a, b and φ are constants.\",\n \"R(n,m) is defined to be a image reflectance function that corresponds to a pixel at location (n,m) of pixel array 700.Using these definitions, the light impinging upon pixel (n,m) is given by: S ( n , m , t ) = R ( n , m ) · I ( t ) = b · R ( n , m ) + a · R ( n , m ) · cos ( 2 π ft + φ ) .\",\n \"Thus, when an image is captured with exposure duration T, the value of the pixel (n,m) will be proportional to the following value: S ( n , m ) = ∫ n Δ n Δ + T S ( n , m , t ) ⅆ t = R ( n , m ) · ∫ n Δ n Δ + T I ( t ) ⅆ t , where is the time period between sampling of two adjacent rows in pixel array 700.The integration of the incident light produces: ∫ n Δ n Δ + T I ( t ) ⅆ t = b · T + a 2 π f [ sin ( 2 π f ( n Δ + T ) + φ ) - sin ( 2 π fn Δ + φ ) ] = b · T + [ a π f sin ( π fT ) ] · cos ( 2 π f Δ n + π fT + φ 2 ) , = b · T + a ′ · cos ( 2 π f ′ n + φ ′ ) , where f ′ = f · Δ a ′ ≡ a π f sin ( π fT ) .\",\n \" φ ′ ≡ π fT + φ 2 Substituting this result provides: S(n,m)=R(n,m)·b·T+R(n,m)·a′·cos(2πf′n+φ).\",\n \"Thus, the resultant signal from pixel (n,m) consists of two components.\",\n \"The first component, R(n,m)·b·T, does not cause flickering artifacts; whereas, the second component can change intensity across rows of pixel array 700.However, as illustrated by the preceding equation, when a′ equals zero, no flickering exists.\",\n \"Therefore, since a′ is a function of exposure duration, it is possible to select such exposure duration that a′ will vanish.\",\n \"Said another way, in some embodiments of the present invention, the flicker frequency is detected, and based on the detected frequency, the exposure time of pixel array 700 is adjusted to reduce flicker.\",\n \"To do this, T is selected to satisfy the following constraint: f·T=k, k= .\",\n \".\",\n \".\",\n \", −2, −1, 0, 1, 2 .\",\n \".\",\n \".\",\n \".\",\n \"To reduce the effect of the frequency components of the S(n,m) columns of each row are summed up to produce row averaged image S(n), where S(n) is described by the following equation: S ( n ) = ∑ m = 0 M - 1 S ( n , m ) = ( b · T + a ′ · cos ( 2 π f ′ n + φ ′ ) ) ∑ m = 0 M - 1 R ( n , m ) = R ( n ) · b · T + R ( n ) · a ′ · cos ( 2 π f ′ n + φ ′ ) .\",\n \"In some embodiments, such a summation is performed by a hardware summing circuit within translation unit 150 where the green and/or any other Bayer components for each row of pixel array 700 are summed.\",\n \"At the end of each row, an eighteen-bit value is sent to memory 118 of control unit 110 using a direct memory access via control bus 101.This same process is repeated for each row of pixel array 700, with each sum being stored to memory 118.Alternatively, in other embodiments, such summing can be accomplished using programmable processor core 112.In some embodiments of the present invention, all rows and pixel values therein are summed.\",\n \"In other embodiments, only a subset of rows including all pixel values therein are summed, while in yet other embodiments, all rows, but only a subset of pixel therein are summed.\",\n \"In yet another embodiment, a subset of the rows and a subset of the pixels therein are summed.\",\n \"From these sum values, S(n) is spectrally analyzed using software executed by programmable processing core 112 to detect the presence of energy corresponding to potential flickering frequency.\",\n \"FIG.\",\n \"27 illustrates a graph 2211 of a typical row average.\",\n \"More particularly, graph 2211 is a plot of the normalized row average 2212 verses the row number 2213 for an image detected by pixel array 700.Various spectral analysis techniques can be used on the sum information to determine a flicker frequency, if any flicker is apparent.\",\n \"For purposes of illustration, operation of flicker detection system 2205 is discussed in relation to a Fourier analysis.\",\n \"More particularly, two energy values, E1 and E2, are determined using the sum data.\",\n \"The two values are calculated as follows: E i = F { S ( n ) } f = f i Δ F { S ( n ) } f = 0 = ∑ n = 0 N - 1 S ( n ) · ⅇ - 2 π f i Δ n ∑ n = 0 N - 1 S ( n ) = ∑ n = 0 N - 1 S ( n ) · cos ( 2 π f i Δ n ) + ∑ n = 0 N - 1 S ( n ) · sin ( 2 π f i Δ n ) ∑ n = 0 N - 1 S ( n ) .\",\n \"In some embodiments, a simple comparison between the energy values E1 and E2 is used to provide an estimation of the flicker frequency.\",\n \"Such estimations, however, can prove unreliable when any ordinary image is the source of the sum information rather than a controlled, constant image.\",\n \"Thus, in various embodiments, a more reliable estimation of the flicker frequency is obtained by calculating a set of E1 and E2 pairs across various frames detected by pixel array 700, and comparing the variance of E1 and E2.This approach provides a number of sampling points across various images and, thus, a more robust estimation of the flicker frequency.\",\n \"Improvement results because there is likely a different phase of the illumination for each frame, which provides various modulations of R(n).\",\n \"After calculation of E1 and E2, corresponding to two possible flickering frequencies, the energy of the present flickering will vary depending on the phase; whereas the energy of the other frequency which depends on the R(n) only, will remain almost the same for all frames.\",\n \"Therefore, the comparison between the standard deviation of E1 and E2 produces reliable results.\",\n \"This procedure is presented in more detail below based on the following definition of variance: V i = 1 K ∑ k = 0 K - 1 ( E i k ) 2 - ( 1 K ∑ k = 0 K - 1 E i k ) 2 , where K is the number of frames that were captured and Eik is the energy value corresponding to frequency fi of the k-th image.\",\n \"To improve the performance, frames can be captured at two or more different exposures that each exhibit flickering.\",\n \"As an example, the following equation describes exposure times creating such a dual flicker: T = ( l + 1 2 ) · 1 f i , i = 1 , 2 ; l = 0 , 1 , 2 , … When both V1 and V2 are below some predefined threshold, no flickering has been detected.\",\n \"Otherwise, the estimated flickering frequency is the frequency that produced larger Vi.\",\n \"FIG.\",\n \"28 illustrates a flow diagram 2241 of a method for detecting and correcting flicker in accordance with the present invention.\",\n \"Following flow diagram 2241, a frame is captured from pixel array 700 (block 2243).\",\n \"The lines of the frame are summed, and E1 and E2 are calculated as previously described (block 2247).\",\n \"Next, it is determined if a sufficient amount of information has been gathered to make a reliable determination of flicker frequency (block 2249).\",\n \"In one embodiment, satisfactory results were obtained when seventeen frames of data are obtained before determining the flicker frequency.\",\n \"If not, more information is gathered by processing additional frames.\",\n \"Alternatively, where sufficient information has been gathered, V1 and V2 are calculated using the gathered information (block 2251).\",\n \"The determined variance value is set equal to the maximum of V1 and V2, and the frequency is set to the frequency fi at which the maximum variance was detected (block 2253).\",\n \"Where the determined variance is less than a predefined threshold level D (block 2257), it is determined that no flicker is detected (block 2259), and the process is repeated to detect a change in conditions that results in flicker.\",\n \"Alternatively, where the determined variance is greater than the a predefined threshold level D (block 2257), a flicker frequency of fi is detected (block 2261) and the exposure duration is constrained to a multiple of 1/fi (block 2263) to correct the detected flicker.\",\n \"In some embodiments, the previously described approach is simplified to reduce computational complexity.\",\n \"More particularly, instead of calculating the exact Fourier component, it is replaced by an approximation of the energy value according to the following equation: E ^ i = ∑ n = 0 N - 1 S ( n ) · SC ( 2 π f i Δ n ) + ∑ n = 0 N - 1 S ( n ) · SS ( 2 π f i Δ n ) ∑ n = 0 N - 1 S ( n ) , where SC ( x ) = sign ( cos ( x ) ) = { 1 , - π 2 ≤ x ≤ π 2 - 1 , π 2 ≤ x ≤ 3 π 2 and SS ( x ) = sign ( sin ( x ) ) = { 1 , 0 ≤ x ≤ π - 1 , π ≤ x ≤ 2 π .\",\n \"However, such a computational reduction can cause degradation in frequency estimation performance.\",\n \"Nevertheless, even after such degradation, satisfactory results of flicker detection and correction can be obtained.\",\n \"Moreover, in some embodiments, only one of the components is used during the estimation process.\",\n \"Such an approach results in insignificant performance degradation which may be overcome by increasing number of considered frames K. In yet other embodiments where greater confidence in the estimated flickering frequency is desired, a more complex approach is utilized.\",\n \"In such approaches, various measures of confidence can be implemented to prevent frequency switches (block 2263) when an incorrect estimate of flicker frequency is detected.\",\n \"Further, in some embodiments, residual flickering (i.e.\",\n \"flickering due to slight variance around the flickering frequency) can be eliminated.\",\n \"More particularly, one such improvement increases the probability of a correct estimation by utilizing more pixel values in the various calculations.\",\n \"Instead of using just a standard deviation of the energy that corresponds to possible flickering frequency to make decision, one may base the estimation on a number, L, of consequent standard deviations.\",\n \"The following equations illustrate such an approach: W 1 = ∑ l = 0 L - 1 V 1 l V 2 l W 2 = ∑ l = 0 L - 1 V 2 l V 1 l C 1 = ∑ l = 0 L - 1 I ( V 1 l > V 2 l ) C 2 = ∑ l = 0 L - 1 I ( V 2 l > V 1 l ) M = max l { ∑ k = 0 K - 1 E 1 k , 1 + E 2 k , l } where I(·) is an indicator function defined below: I ( Condition ) = { 1 , if Condition is true 0 , otherwise , and Vxl,Exk,l denotes the l-th standard deviation and the energy of k-th frame in l-th run, respectively.\",\n \"Note that the total number of frames that should be acquired prior the decision is K·L.\",\n \"The decision is made based on the following set of rules: 1.If M is below a predefined flickerless threshold D0 no flicker artifact was detected.\",\n \"2.If W1>D1∩C1>D2, flicker at f1 was detected.\",\n \"3.If W2>D1∩C2>D2, flicker at f2 was detected.\",\n \"4.Otherwise, the information is ambiguous and no decision is taken.\",\n \"This method reduces drastically the probability of both misdetection and incorrect detection.\",\n \"However, the delay and probability of false alarm are increased, which can be less critical in some applications.\",\n \"Further, to prevent the situation when occasional wrong frequency estimation changes the exposure to a value that leads to flicker artifact, some embodiments include additional mechanisms to reduce this possibility.\",\n \"One such approach for avoiding this situation includes an additional counter, referred to as a confidence counter.\",\n \"The confidence counter is initialized to zero.\",\n \"Whenever flicker at a first frequency, f1, is estimated the confidence counter is increased by one for each estimation until a limit, B, is reached.\",\n \"If estimated flicker frequency is a second frequency, f2, the confidence counter is decreased by one for each estimation until a limit, negative B, is reached.\",\n \"In a case of ambiguity, no change is made to the confidence counter.\",\n \"When the confidence counter is positive, the global estimation is f1 and exposure is limited to multiple of 1/f1.When the confidence counter is negative, the global estimation is f2 and exposure is limited to multiple of 1/f2.If the confidence counter is zero previous estimation is used.\",\n \"This reduces the possibility of spurious data creating an unstable exposure time.\",\n \"Parallel Interface Parallel interface 190 produces the video output of imaging device 100.parallel interface 190 includes a ten-bit parallel data bus, a vertical frame signal, a clock, and a qualifying signal.\",\n \"Parallel interface 190 can also produce a serial output selectable by programming a register.\",\n \"When operating in a parallel mode, imaging device 100 outputs a video image in a parallel format.\",\n \"The parallel format includes a clock signal along with data signals and qualifying signals each synchronized to the clock signal.\",\n \"In some embodiments of imaging device 100, the effective edge of the clock signal (i.e., rising or falling edge) is programmable.\",\n \"The data signals include a ten-bit wide data bus carrying either the Bayer grid data, or the YUV 4:2:2 output data, where the type of data is determined by monitoring a programmed register.\",\n \"The eight-bit YUV data is presented on the most significant data signals (i.e., bits three through ten of the ten-bit bus).\",\n \"On each effective edge of the clock, a single data byte is transferred on the data bus where a horizontal qualifying signal is asserted.\",\n \"The horizontal qualifying signal is asserted only during valid image data, or to qualify optional start-of-line and end-of-line markers.\",\n \"A vertical qualifying signal is asserted on the effective edge of the clock to mark the beginning of the first line in a new image frame.\",\n \"The duration of the vertical qualifying signal is programmable via a register implemented as part of imaging unit 130.FIG.\",\n \"29 illustrates the vertical qualifying signal, VALIDV, and the horizontal qualifying signal, VALIDH, in relation to a pixel array where the active image portion of the array is designated.\",\n \"As illustrated, VALIDV is a vertical synchronization signal whose effective edge (programmable by setting a register value) marks the beginning of the first line in a new image frame.\",\n \"FIG.\",\n \"30 illustrates the signals in relation to data signals referred to a valid data, and FIGS.\",\n \"34 and 35 illustrate a bitwise view of the signals in relation to Bayer and YUV data, respectively.\",\n \"As illustrated, when sending out the data in Bayer-grid format, a “RED” line will always be the first line in the frame.\",\n \"When sending out the data in YUV output format, the order of the color components is programmable via a register value.\",\n \"Control unit 110 controls the frame rate output via parallel interface 190 via a C_outframe signal.\",\n \"Via the signal, control unit 110 enables or disables the output of a full frame.\",\n \"When the signal is deasserted, VALIDH and VALIDV signals are stuck at an inactive state and the data signals hold the background value throughout the disabled frame time.\",\n \"By deasserting C_outframe, a frame of data can be skipped as illustrated in FIG.\",\n \"33.The type of data output via parallel interface 190 is programmable.\",\n \"When Bayer data is output, the data volume is an 8/9/10 bit word.\",\n \"If data markers are added, eight more bytes per line must be taken into consideration in the image buffer that is allocated for the application.\",\n \"There are two types of BAYER data lines: red lines and blue lines.\",\n \"Red lines carry green and red Bayer components.\",\n \"The green and red data bytes are interleaved where the even numbered bytes (0, 2 to 2n) are green, and the odd numbered bytes (1, 3 to 2n+1) are red.\",\n \"On the other hand, blue lines carry blue and green Bayer components, where the components are interleaved with the even numbered bytes (0, 2 to 2n) being blue, and the odd numbered bytes (1, 3 to 2n+1) being green.\",\n \"When sending Bayer data out on parallel interface 190, a red line is the first line in the frame.\",\n \"When YCrCb 4:2:2 data is output, the data volume is two bytes per pixel and no extra pixels are transmitted beyond the CIF/QCIF size.\",\n \"If data markers are added, eight more bytes per line must be taken into consideration in the image buffer that is allocated for the application.\",\n \"The data in the YCrCb 4:2:2 output format is ordered in several configurations, according to the setting in a programmable color register.\",\n \"The following illustrates four possible color orders: 00—SOL, Y0, U0, Y1, V0, Y2, U2, Y3, V3, to Yn-2, Un-2, Yn-1, Vn-2, EOL 01—SOL, Y0, V0, Y1, U0, Y2, V2, Y3, U3, to Yn-2, Vn-2, Yn-1, Un-2, EOL 10—SOL, U0, Y0, V0, Y1, U2, Y2, V3, Y3, to Un-2, Yn-2, Vn-2, Yn-1, EOL 11—SOL, V0, Y0, U0, Y1, V2, Y2, U3, Y3, to Vn-2 Yn-2, Un-2, Yn-1, EOL In some embodiments, extra data markers are inserted at the beginning and end of each image line.\",\n \"This insertion is optional and can be selected via a programmable register.\",\n \"The following are four examples of data marker types: Start of First Line marker—This marker precedes the data of the first line in a frame.\",\n \"Start of Line Marker—This marker precedes the data of each line in a frame, except for the first line.\",\n \"When Bayer output mode is selected, bit #5 of the marker content differentiates between a “red” line and a “blue” line (0 and 1 respectively).\",\n \"End of Last Line Marker—This marker immediately follows the data of the last line in a frame.\",\n \"End of Line Marker—This marker immediately follows the data of each line in a frame, except for the last line.\",\n \"In addition to outputting data in a parallel mode, parallel interface 190 can format the data for serial output.\",\n \"The Transmitted data size for serial output mode is eight bits per transmitted “word”.\",\n \"Bayer output provides eight bits per pixel, and YUV output is sixteen bits per pixel.\",\n \"The bytes are transmitted with the most significant bits first.\",\n \"During a continuous transfer, the most significant bit of byte N immediately follows the least significant bit of byte N−1.In some embodiments, imaging device 100 is integrated into systems where the Host is pin limited.\",\n \"For example, in one particular embodiment, the host is a Digital Signal Processor capable of receiving video via a two or three pin interface.\",\n \"In such systems, imaging device 100 can be configured to provide video output via a serial interface.\",\n \"In some embodiments the serial mode provided by imaging device 100 relies on elements consistent with ITU-RBT.656-4 standard.\",\n \"ITU-RBT.656-4 standard recommendation is available from the International Telecommunication Union, the entirety of which is incorporated herein by reference for all purposes.\",\n \"In one particular embodiment, imaging device 100 can be configured to operate in different serial output modes: a clock-qualify mode and a synchronous serial interface (“SSI”) mode.\",\n \"Such functionality provides an ability to function in low cost, reduced pin-count environments.\",\n \"In contrast to ITU-RBT.656-4 standard, in embodiments of the present invention where data markers are used, the EAV and SAV markers are different.\",\n \"In particular, the second most significant bit (i.e.\",\n \"bit six) of the markers is a special frame marker used to indicate the start of the first line of a frame and the end of the last line of a frame.\",\n \"Further, bit five indicates the Bayer color, where a logical one indicates blue and a logical 0 indicates red.\",\n \"In the serial output modes, either two or three pins of the thirteen-bit parallel video interface 192 are utilized to output the serial video data.\",\n \"Further, in each of the serial output modes, the frequency of the clock, DSCLK 2920, used to output the serial data is eight times (“8×”) that of a clock, CLK, used to output parallel video data via parallel video interface 192.In some embodiments, DSCLK 2920 is provided via an external pin to imaging device 100.In such cases, the input clock source is internally divided to provide clocking useful for operating parallel video interface 192 when both parallel and serial video data are being provided from imaging device 100.In other embodiments, an internal phase lock loop is used to generate the various output clocks for use in relation to both the serial and parallel output modes of imaging device 100.In operation, parallel video data is received from translation unit 150.Where a serial output mode is selected via a programmable register, the parallel data is shifted out via the designated serial pins of parallel video interface 192 using the 8× clock.\",\n \"This shifting occurs by transmitting the most significant bit of any parallel data first and following in succession with bits of decreasing significance.\",\n \"Clock-qualify mode is a two wire interface with clock and output data signals, DSCLK 2920 and DSDATA 2940.Imaging device 100 acts as a master of the interface.\",\n \"As such, imaging device 100 activates DSCLK 2920 when there is valid data to be sent out via DSDATA 2940.FIG.\",\n \"34 illustrates the clock-qualify mode where data can be output at two rates represented by DSCLK/2 2950 and DSCLK 2920, both being synchronized to CLK of imaging device 100.As illustrated, DSDATA 2940 is provided by imaging device 100 on the rising edge of DSCLK/2 2950 and latched by the host on the falling edge of DSCLK/2 2950.It should be recognized that twice the transfer rate is possible where the same approach is used in relation to DSCLK 2920.Using the approach, eight bits are transferred from the most significant bit to the least significant bit.\",\n \"In some embodiments, one deasserted cycle of DSCLK 2920 or DSCLK/2 2950 indicates a word boundary, while in other embodiments, word boundaries are contiguous allowing for a continuous transfer of data based on a continuing period of DSCLK 2920 or DSCLK/2 2950.SSI mode is a three wire interface, with clock (DSCLK 2920), output data (DSDATA 2940) and frame signals (DSFRAME 2930).\",\n \"Imaging device 100 acts as a master of the interface, producing all three signals.\",\n \"FIG.\",\n \"35 illustrates the operation of SSI mode with DSCLK 2920, DSFRAME 2930, and DSDATA 2940.As illustrated, DSFRAME 2930 is asserted by imaging device 100 prior to a rising edge of DSCLK 2920.The host accepts the asserted DSFRAME 2930 on a rising edge 2910 of DSCLK 2920.On the eight following rising edges of DSCLK 2920, one of the eight bits of a byte transfer are provided by imaging device 100 as DSDATA 2940 to a receiving host.\",\n \"The bits are provided with the most significant bit of the by first followed sequentially by bits of decreasing significance.\",\n \"Testability Unit Imaging device 100 can additionally include various mechanisms for performing tests of the device.\",\n \"Using one such mechanism, a built-in-self-test (BIST) is implemented to facilitate production testing of ADC 134 and the various DAC circuitry.\",\n \"The BIST circuit creates a sequence of values that are passed to ADC 134 after application of the gain and two offset parameters as previously discussed.\",\n \"The output from sensor 132.With this canned value presented to ADC 134, the outputs of ADC 134 is checked against a known output.\",\n \"If the output falls within a programmable tolerance level, imaging device 100 is acceptable.\",\n \"If not, imaging device 100 is rejected.\",\n \"In addition, a BIST mechanism for sensor 132 is implemented.\",\n \"A minimum and maximum value is defined for each color within sensor 132.Sensor 132 is illuminated by a monochrome color, and the resulting image is tested.\",\n \"The value of each pixel is checked.\",\n \"At the end of the frame, three values can be read per each color.\",\n \"The number of pixels below the minimum value, the number of pixels above the maximum value, and the number of pixels falling within the minimum and maximum values is determined.\",\n \"Additionally, BIST mechanisms are implemented to test various RAMs and registers within imaging device 100 and to return pass or fail results.\",\n \"The invention has now been described in detail for purposes of clarity and understanding.\",\n \"However, it will be appreciated that certain changes and modifications may be practiced within the scope of the appended claims.\",\n \"Accordingly, it should be recognized that many other systems, functions, methods, and combinations thereof are possible in accordance with the present invention.\",\n \"Thus, although the invention is described with reference to specific embodiments and figures thereof, the embodiments and figures are merely illustrative, and not limiting of the invention.\",\n \"Rather, the scope of the invention is to be determined solely by the appended claims.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10474275"}}},{"rowIdx":794,"cells":{"text":{"kind":"list like","value":[["Surface profile measurement","An imaging system scans a light source output over an object to be imaged, and receives the reflected signal in a multi-region light detector, wherein different regions of the object to be imaged are imaged onto different regions of the detector by the receiving optics, and wherein different regions of the light detector can be actuated separately.","The light source scanning is synchronised with the actuation of the light detector so that a region of the detector imaging the region of object being illuminated by the light source is actuated.","The time of flight of light signals from the light source to the actuated portion of the detector is calculated for all scanning directions, to enable a three dimensional image of the object to be constructed."],["1.An imaging system comprising: a light source; means for scanning the light from the light source over an object to be imaged; stationary receiving optics for receiving light reflected from the object to be imaged; a multi-region light detector for detecting light received from the receiving optics, wherein different regions of the object to be imaged are imaged onto different regions of the detector by the receiving optics, and wherein different regions of the light detector can be actuated separately; control means to synchronise the light source scanning and the actuation of the light detector so that a region of the detector imaging the region of the object being illuminated by the light source is actuated; and processing means for measuring the time of flight of light signals from the light source to the actuated portion of the detector for all scanning directions.","2.An imaging system as claimed in claim 1, wherein the or each light source is a pulsed laser or light emitting diode or diodes.","3.An imaging system as claimed in claim 1, wherein the light source is a laser or light emitting diode or diodes having a modulated output.","4.An imaging system as claimed in any preceding claim, wherein the means for scanning comprises: a reflector for directing the light from the light source to an object to be imaged; and a drive mechanism for controlling movement of the reflector for scanning the light from the light source over the object to be imaged.","5.An imaging system comprising: a plurality of light sources, each light source being directed to an object to be imaged; stationary receiving optics for receiving light reflected from the object to be imaged; a multi-region light detector for detecting light received from the receiving optics, wherein each light source is imaged by the receiving optics onto a different region of the light detector, wherein the different regions of the light detector can be actuated separately; control means to synchronise the operation of individual light sources and the operation of the light detector so that the region of the detector imaging the region of the object being illuminated by an operating light source is actuated; and processing means for measuring the time of flight of light signals from the operating light source to the actuated portion of the detector for all scanning directions.","6.An imaging system as claimed in claim 5, wherein each light source comprises a pulsed laser or light emitting diode or diodes.","7.An imaging system as claimed in claim 5, wherein the light source is a laser or light emitting diode or diodes having a modulated output.","8.An imaging system comprising: a light source directed to an object to be imaged; stationary receiving optics for receiving light reflected from the object to be imaged; a multi-region light detector for detecting light received from the receiving optics; control means to synchronise the operation of the light source and the operation of the light detector so that a region of the detector is actuated synchronously with an allocated operation of the light source; and processing means for measuring the time of flight of light signals from the light source to the actuated portion of the detector.","9.An imaging system as claimed in claim 8, wherein all regions of the light detector are actuated in parallel.","10.An imaging system as claimed in claim 8, wherein different regions of the light detector can be actuated separately.","11.An imaging system as claimed in claim 10, wherein different sub-regions of the light detector are actuated in sequence, a different operation of the light source being allocated to each sub-region.","12.An imaging system as claimed in claim 10 or 11, wherein each sub region comprises a row or column of detector pixels of a pixel array.","13.An imaging system as claimed in any one of claims 8 to 12, wherein the light source comprises a pulsed laser or light emitting diode or diodes.","14.An imaging system as claimed in any one of claims 8 to 12, wherein the light source is a laser or light emitting diode or diodes having a modulated output.","15.An imaging system as claimed in any preceding claim, wherein the light detector comprises a photodiode array.","16.An imaging system as claimed in claim 15, wherein the photodiode array is operable in two modes, a first mode in which light signals from all photodiodes in the array are read out in sequence, and a second mode in which light signals from selected photodiodes or sub-groups of photodiodes in the array are read out in a sequence.","17.An imaging system as claimed in claim 15 or 16, wherein the processing means is integrated onto the same substrate as the photodiode array.","18.An imaging system as claimed in any one of claims 15 to 17, wherein multiple channels of time of flight measurement circuitry are integrated onto the same substrate as the photodiode array and configured to process signals from different actuated portions of the photodiode array in parallel.","19.An imaging system as claimed in any preceding claim, wherein the processing means uses cross-correlation for measuring the time of flight of light signals from the light source to the actuated portion of the detector.","20.A method of obtaining an image of an object comprising: scanning a light source signal over the object by directing a light source output in a plurality of scanning directions in sequence, and detecting reflected light received from the object using a two dimensional light detector array; determining the regions of the light detector array which are illuminated for each scanning direction; scanning the light source signal over the object again, and detecting reflected light received from the object using only the determined regions of the light detector array; calculating the time of flight of light pulses from the light source to the detector for each scanning direction; and obtaining a 3D profile from the time of flight calculations.","21.A method as claimed in claim 20, wherein the scanning of the light source is synchronised with the actuation of selected regions of the light detector array.","22.A method as claimed in claim 20 or 21 wherein the light source signal comprises a pulsed laser output or output of a light emitting diode or diodes.","23.A method as claimed in claim 20 or 21, wherein the light source signal comprises a modulated output of a laser or light emitting diode or diodes.","24.A method as claimed in claim 23, wherein calculating the time of flight uses cross-correlation."],["The invention relates to apparatus for measuring the surface profile of a sample, and in particular relates to non-contact 3D surface profile measurement systems.","The non-contact measurement of three-dimensional (3D) objects to extract data regarding their physical form and location in space is a topic which has excited much research.","Many techniques have been developed to suit the distance to the object, the precision with which the object's features need to be measured and so on.","One common technique is to illuminate the remote object with a light source of known pattern; so called “structured illumination”.","A camera is located some distance away from the structured light source and arranged to collect an image of the projection of the structured light pattern onto the surface of the remote object.","FIG.","1 shows this known profile measurement using structured lighting.","A structured light source 10 projects light onto the object 12, and the reflected light is captured by a camera 14 having a field of view 15 which covers the object 12.An image processor 16 derives the 3D profile data 18 by analysis of the deformation of the structured light pattern, which is representative of the distance to the remote object.","The 3D nature of the remote object causes the structured light pattern in the image captured by the camera to be deformed.","From knowledge of the physical separation of the camera from the light source (the baseline 20) and trigonometry, it is possible to compute the 3D surface profile of the remote object from the deformation of the image of the structured light pattern.","A benefit of this system is that if a video camera is used to capture the image, then with sufficient processing, the 3D surface profile can be measured at video frame rates (50 or 60 times per second).","As this system works on trigonometric principles, the depth accuracy is related to the baseline of the system and the resolution of the sensor.","As a result, practical considerations tend to limit the application of such systems to remote objects no more than a few metres away.","In addition, as the baseline is lengthened, the angle between the incident light pattern and the line of sight of the camera becomes more obtuse and shadows cast by features in the remote object can obscure the 3D profile measurement.","Alternative techniques rely on scanning a device which measures the distance to a remote point; e.g.","a laser rangefinder (LRF) or interferometer, using a remote controlled pan and tilt unit.","An example of such a system is illustrated in FIG.","2.A distance measuring device such as a laser rangefinder 30 measures the distance to a single point on the object, and is controlled by a pan-and-tilt unit 32.Control electronics 34 under the control of a computer causes the pan and tilt unit 32 to scan the line of sight of the distance measuring device 30 across the object to be measured to build up a 3D matrix of the azimuth, elevation and distance to the remote object.","This matrix of numbers represents the 3D surface profile of the remote object measured in polar co-ordinates from the axis of rotation of the pan and tilt unit.","Some corrections may be applied to the data if the axes of rotation of the pan and tilt unit are not co-incident with the line of sight of the measuring device.","The resultant 3D surface profile 18 can be transformed to other co-ordinate systems using known mathematical techniques.","This technique is analogous to radar systems where the time of flight of a scanned micro-wave radio signal is used to determine the distance to one or more remote objects.","This scanned LRF approach does not suffer from the range limitations of the structured light approach, but is relatively expensive to implement because high precision and costly components are required for good accuracy.","In addition, because the whole mass of the distance measuring device is scanned, achieving scanning at rates sufficient to give “real time” 3D image data is problematic.","To overcome this difficulty, systems have been built using a scanned mirror or mirrors in front of the LRF.","For example PCT/US97/06793 describes a complex system using two orthogonal mirrors to scan the LRF beam horizontally and vertically so as to cover a field of view of approximately 40°×40°.","This approach enables the LRF to remain static.","However, the mirror/mirrors must be sufficiently large to encompass both the laser beam and lens aperture of the receiving system without vignetting or optical crosstalk between the emitted laser beam and received beam and so such mirror scanned systems still remain expensive.","Hybrid approaches, where the distance measuring device is scanned along a line in one direction and the remote object is rotated have been used to create a map of the complete surface of a the remote object.","However, these techniques are not appropriate for all targets.","Other workers have sought to overcome these problems by scanning the laser beam whilst keeping the detector static, as described in GB 2 286 495.With this approach, the only moving part of the system is a scanned mirror which only need be sufficiently large to steer the laser beam.","This avoids the high cost of a precision motorised pan and tilt head and enables a high scan rate.","Furthermore, because the laser and receiving optical paths can be kept completely separate there is no risk of optical crosstalk.","However, this approach requires the field of view of the receiving optical system to encompass the whole of the remote object.","As a result, the optical system will collect illumination from the whole of the field of view.","This generates a background offset signal level in the photodetector of the optical system.","Whilst this offset level can be compensated for in the signal detection process, the shot noise inherent in the offset signal level significantly degrades the ratio of detected laser signal level to noise and hence reduces the maximum range of the system.","To overcome this significant disadvantage some very complex schemes have been proposed.","One such is as described in U.S. Pat.","No.","5,682,229 where the reflected light from a scanning laser is focussed onto a stationary detector assembly which uses a series of complex fibre bundles to transfer the collected light to 128 separate avalanche photodiodes and associated time of flight measuring circuitry.","It will be appreciated that the complexity and use of relatively expensive components such as coherent fibre-optic assemblies and multiple discrete avalanche photodiodes makes such approaches expensive to implement.","According to the invention, there is provided an imaging system comprising: a light source; means for scanning the light from the light source over an object to be imaged; stationary receiving optics for receiving light reflected from the object to be imaged; a multi-region light detector for detecting light received from the receiving optics, wherein different regions of the object to be imaged are imaged onto different regions of the detector by the receiving optics and wherein different regions of the light detector can be actuated separately; control means to synchronise the light source scanning and the actuation of the light detector so that a region of the detector imaging the region of the object being illuminated by the light source is actuated; and processing means for measuring the time of flight of light signals from the light source to the actuated portion of the detector for all scanning directions.","The direction of scanning and the associated measured time of flight can be used to build up a three dimensional image of the object.","The arrangement of the invention provides a simple structure with few moving parts.","The means for scanning may comprise a reflector for directing the light from the light source to an object to be imaged, and a drive mechanism for controlling movement of the reflector for scanning the light from the light source over the object to be imaged.","According to a second aspect of the invention, there is provided an imaging system comprising: a light source directed to an object to be imaged; stationary receiving optics for receiving light reflected from the object to be imaged; a multi-region light detector for detecting light received from the receiving optics; control means to synchronise the operation of the light source and the operation of the light detector so that a region of the detector is actuated synchronously with an allocated operation of the light source; and processing means for measuring the time of flight of light signals from the light source to the actuated portion of the detector.","This aspect enables an imaging system to be made with no moving parts, using a multi-region light detector.","All regions of the light detector can be actuated in parallel, for high speed image capture.","However, different regions of the light detector can preferably be actuated separately.","This enables different sub-regions of the light detector to be actuated in sequence, a different operation of the light source being allocated to each sub-region.","For example, each sub region may comprises a row or column of detector pixels of a pixel array.","According to a third aspect of the invention, there is provided an imaging system comprising: a plurality of light sources, each light source being directed to an object to be imaged; stationary receiving optics for receiving light reflected from the object to be imaged; a multi-region light detector for detecting light received from the receiving optics, wherein each light source is imaged by the receiving optics onto a different region of the light detector, wherein the different regions of the light detector can be actuated separately; control means to synchronise the operation of individual light sources and the operation of the light detector so that the region of the detector imaging the region of the object being illuminated by an operating light source is actuated; and processing means for measuring the time of flight of light signals from the operating light source to the actuated portion of the detector for all scanning directions.","According to this aspect, the need to scan the light source output over the object to be imaged is again avoided.","In each of these aspects, the or each light source may comprise a pulsed laser or light emitting diode or diodes, and the output may be modulated, with cross correlation techniques used for the time of flight measurement.","In each of these aspects of the invention, the light detector preferably comprises a photodiode array, which can be implemented at low cost.","The photodiode array may be operable in two modes; a first mode in which light signals from all photodiodes in the array are read out in sequence, and a second mode in which light signals from selected photodiodes or sub-groups of photodiodes in the array are read out in a sequence.","The invention also provides a method of obtaining an image of an object, comprising: scanning a light source signal over the object by directing a light source output in a plurality of scanning directions in sequence, and detecting reflected light received from the object using a two dimensional light detector array; determining the regions of the light detector array which are illuminated for each scanning direction; scanning the light source signal over the object again, and detecting reflected light received from the object using only the determined regions of the light detector array; calculating the time of flight of light pulses from the light source to the detector for each scanning direction; and obtaining a 3D profile from the time of flight calculations.","Examples of the invention will now be described in detail with reference to the accompanying drawings, in which: FIG.","1 shows a first known 3D imaging system; FIG.","2 shows a second know 3D imaging system; FIG.","3 shows a simplified example of the photodiode array for use in the system of the invention; FIG.","4 shows an imaging system of the invention, using the photodiode array of FIG.","3; FIG.","5 shows a modification to the photodiode array; FIG.","6 shows how pulsed light sources can be used in the imaging system of the invention.","The invention provides a 3D imaging system, with a stationary receiving optical system.","The invention uses a two dimensional detector, which will first be described with reference to FIG.","3.FIG.","3 is a simplified schematic of a photodiode array for use in an imaging system of the invention, in a 2×2 format.","The device consists of an array of photodiode pixels 60, each of which comprises a photodiode (PD11 to PD22) and associated transistor (TR11 to TR22), which are configured and drive to act as analogue switches.","For standard video imaging applications, the device is operated in an integration mode where incident illumination is focussed upon its surface.","The incident illumination generates charge within each photodiode by the photoelectric effect.","During this integration period, connections X1, X2, Y1 and Y2 are all held low so that all transistors are off and the photodiodes are electrically isolated.","The photo-generated charge then accumulates in each photodiode and is stored on the self-capacitance of the photodiode.","Once sufficient photocharge has been collected, the device is readout as follows.","Input X1 is taken to a high potential so that TR1 is turned on thereby allowing charge to flow between the column and a charge sensitive amplifier 62.Then input Y1 is pulsed high for addressing a row of pixels, turning TR11 on allowing and the photo-generated charge stored on photodiode PD11 to flow through TR11 and TR1 to the output amplifier 62 where the charge is converted to a voltage.","This creates an output signal whose amplitude is proportional to the level of charge stored on PD11 and hence the level of light incident on TR11.After the self capacitance of PD11 has been discharged, input Y1 is taken low and input Y2 is taken high, allowing the stored charge on PD12 to be readout.","In this way, a column of pixels is read out in turn.","After all the charge collected by PD12 has been discharged, Y2 is taken low and X2 is taken high to allow PD21 and PD22 (the pixels in the next column) to be readout sequentially by pulsing Y1 and Y2 in the manner described above.","It can be seen that this process allows the 2×2 array to be scanned and an electrical signal that is the analogue of the incident illumination generated.","In normal operation, larger numbers of photodiodes are used, e.g.","256×256, to increase resolution.","Often the readout sequence and sensor scanning are arranged to generate a standard video signal.","In addition, it may be noted that the basic structure described here has been simplified for the purpose of describing the proposed invention.","Practical X-Y addressed photodiode arrays are generally fabricated as single complementary metal oxide semiconductor (CMOS) large scale integrated circuits (LSI) which include many refinements such as on-chip clock circuitry to generate the pulse sequences for electrodes X1 to Xn and Y1 to Yn.","On-chip and additional pixel and/or column level circuitry improve amplification and detection of the photo-charge.","These known refinements can be applied with advantage to the 3D profile measurement principle described here.","For 3D profile measurement, the inventor has realised that such X-Y addressed photo-diode arrays can be utilised not in an integrating mode, but as a multiplexer synchronised with the laser scanning and pulsing so that only the individual photodiode receiving the reflected image of the laser spot on the remote object is addressed.","FIG.","4 shows a system using the photodiode array in this synchronised mode of operation.","A sequentially pulsed laser beam output from a laser 40 is scanned by known means to illuminate a point on the surface of the remote object 12.To minimise size and cost, the laser scanning system 44 may be implemented in a number of ways including using electromagnetically or piezo-electrically scanned mirrors 42 or by mounting a laser chip on a micro-machined silicon or compact piezo electric structure.","An image of the laser spot is formed on the surface of the X-Y addressed photodiode array 70 by the stationary, receiving optical system 46.A narrow band-pass optical filter 50 may be used to reject all wavelengths except those near the laser wavelength and optimise discrimination of the laser pulse against the background illumination.","The control electronics apply logic level signals to the relevant X and Y control lines of the X-Y addressed array so that the photodiode illuminated by the image of the laser spot is connected to the pre-amplifier and time of flight detection electronics 56.In particular, the reflected laser pulse is captured by this photodiode and the resultant electrical signal routed to an electrical pulse detector 54 and TOF (time of flight) measurement circuitry 56.This computes the TOF of the laser pulse to the spot on the remote object and back to the photodiode on the X-Y addressed array and hence distance from the remote object to the X-Y addressed array.","This process is then repeated for many points over the surface of the remote object to measure the surface profile of the remote object.","If the image formed of the laser spot is larger than a single pixel then the control electronics can cause the detector to address a group of adjacent photodiodes (e.g.","a 2×2 s-array of photodiodes) in parallel to optimise collection and detection of the laser energy.","It can be seen that because only one or a small number of the photodiodes is connected to the receiving amplifier and time of flight electronics at any one time, the background offset signal will be limited to that generated by the part of the field of view focussed onto the individual photodiode/photodiode group, rather than from the whole of the field of view of the optics as for the system described above.","For example, if the laser beam is arranged to sequentially scan 100×100 equispaced points within the optical field of view, then the background signal level collected by each photodiode/photodiode group will be nominally reduced by 10,000 times in comparison to the simple system described above, which confers substantial benefits on system performance.","It may be noted that a simplified version of this system can be used with a linear photodiode array to measure a 2D, rather than 3D surface profile.","The performance of the system can be substantially improved by replacing the pulsed laser source 40 with a modulated laser source and the pulse discriminator 56 by a cross-correlation system.","Such systems are known, for example, from DE19949803 to Denso Corp and PCT/GB/04968.In particular, the system may include a signal source such as a laser for supplying a modulation signal and a transmission system connected to the signal source for transmitting a transmitted optical signal modulated by the modulation signal.","The modulation signal may be, for example a maximal length sequence.","A reception system is then provided for receiving a reflected and delayed version of the transmitted signal, and a cross-correlator for obtaining the time delay.","The cross correlator can be arranged to determine, at a coarse resolution, the time delay of the modulation signal needed to maximise the correlation between the time delayed modulation signal and the received signal.","The cross correlator can then determine, at a finer resolution than the coarse resolution, the correlation between the modulation signal and the received signal as a function of the time delay of the modulation signal with respect to the received signal in a smaller time delay range around the determined time delay.","A measure of distance is calculated from the time delay of the modulation signal needed to maximise the correlation between the time delayed modulation signal and the received signal.","The cross-correlator can be implemented digitally.","In this way, for a given laser peak power, greater energy can then be delivered to the remote object which improves the signal to noise ratio and hence maximum range of the system.","This oversampling approach enables the distance resolution of the system to be improved, and the efficient signal processing method using coarse and fine cross-correlators minimises the processing power needed.","In a preferred embodiment, the X-Y sensor, time of flight measurement system and control electronics are fabricated on a single integrated circuit to minimise manufacturing cost.","The photodiodes can be manufactured and operated as avalanche photodiodes to provide signal amplification by the avalanche effect, prior to signal detection.","The time of flight measurement approach used may be as described above in connection with FIG.","4 or other higher performance schemes described above.","X-Y Sensors such as described here can suffer from image lag which arises because a) the photo charge generated by incident light upon a pixel is partitioned between the photodiode's intrinsic capacitance and the capacitance of the row and column electrodes resulting in some charge being left on the row and column electrode capacitances after each pixel is read out and b) a load transistor may be provided to discharge the photodiode capacitance and bias the amplifier, but the effective resistance of this load transistor increases with declining current increasing the time constant of the system at low light levels.","In a preferred embodiment, this effect may be minimised by configuring the receiving amplifier as a transimpedance amplifier whilst the photodiode array is used in a multiplexer mode.","The virtual earth offered by the transimpedance amplifier's input provides a low impedance path through which the photo generated current can flow, reducing the charge partitioning effect and improving system bandwidth.","This approach also provides a means to bias the photodiodes during readout.","In an optimal development of the preferred embodiment, a transimpedance amplifier stage is provided for each pixel to drive the multiplexing circuitry of the array.","The laser scanning pattern will often be a repeating pattern arranged to cover the optical field of view whilst providing adequate time resolution to measure the position of moving objects in the field of view.","The pattern is typically arranged as a conventional raster scan for ease of display on conventional monitors.","However, it can be seen that other patterns may be used.","One useful pattern is a spiral scan pattern where by controlling the velocity of the laser scan, increased spatial resolution may be achieved in the centre of the scan whilst still maintaining a low spatial resolution to detect objects appearing at the periphery of the scan.","For those applications where it is necessary to monitor the 3D motion of a few specific objects within the field of view, the scan can be controlled adaptively to track the objects and ignore the rest of the field of view.","This approach can increase the temporal resolution for the tracked objects.","An important benefit of the approach described here is that the X-Y addressed array can still be operated in an imaging mode, rather than a multiplexed time of flight detector mode.","This can be achieved simply by returning the sequence of pulses applied to the X-Y addressed array to a conventional video scanning sequence.","This has several significant benefits.","First, for optimal performance, it is important that only the photodiode or local group of photodiodes receiving the image of the laser spot at any point in time are addressed; i.e.","that the laser scanning and the photodiode array scanning are synchronised.","This would normally require precise calibration of the scanner and optical system.","However, if the laser is scanned whilst the sensor is in an imaging mode, an image of the laser path can be collected by the control electronics.","This image can be used to determine the precise path of the laser beam image on the surface of the photodiode array and hence set up the correct addressing sequence for the X-Y addressed array and/or laser pulsing sequence, to ensure synchronisation in the multiplexed time of flight detector mode.","Thus, the normal addressing mode is used as a calibration stage for the higher performance multiplexing mode.","In effect, the system can be self calibrating which is a major benefit for systems which have to operate over large temperature ranges.","Secondly, if the laser is turned off and the X-Y addressed photodiode array is operated in an imaging mode, then the system can be used as a conventional imaging device; i.e.","a video or still camera to provide additional information regarding the remote object.","Third, because the same detector is used to capture a 3D surface profile and standard image, the registration between the standard image and 3D data is near perfect.","This enables beneficial combinations of conventional and imaging and 3D profile measuring modes to be used.","For example, by toggling the system between capturing a 3D scan and a conventional image scan both a video image sequence and 3D sequence to be captured and overlaid on one another.","This approach is particularly beneficial for collision avoidance or intruder detection.","If the 3D image shows that an object within the field of view is too close or is on a path likely to lead to a collision, the corresponding part of the conventional image can be coloured or caused to flash to draw, attention to the problem.","Another benefit of capturing perfectly registered 3D and conventional image data is that image processing methods can be applied simultaneously to both sets of data, using the optimal method to extract important features from each image and combining the feature data to yield more information about objects in the field of view.","For example, the 3D data can be used to determine the orientation and size of an object in the field of view.","This data can then be used to select, or generate from a 3D solid model of a known object, an appropriate pattern for pattern matching and object/target recognition.","It can be seen that the benefits of acquiring perfectly registered 3D and conventional image data are many.","Where higher scanning rates are required, the system described above can be improved by modifying the optics of the laser to illuminate a line, rather than a point, of the remote object and modifying the structure of the X-Y addressed array to permit a row of photodiodes to be connected in parallel to multiple time of flight measurement processors; i.e.","a Y scanned, X parallel output array.","One implementation of the revised photodiode array structure is shown for a 2×2 array in FIG.","5.Each column switch transistor has been replaced by an amplifier 80 and time of flight measurement (TOF) circuitry comprising a pulse detector 82, counter 84 and latch 86.All counters 84 are clocked at the same frequency by a common clock input 88.The laser scanning system is set to illuminate a strip of the remote object.","The image of the illuminated strip is focussed onto row N of the photodiode array.","The laser is pulsed on, the TOF counters 84 are reset to zero and the control electronics takes the relevant Y electrode (Y1,Y2) high to connect all the photodiodes along the row to the corresponding amplifier & TOF circuitry.","As each photodiode receives the reflected pulse of laser light, the pulse detector causes the corresponding counter 84 to stop counting.","Once all reflected pulses have been received, the number held on each counter 84 is latched into the latches 86 and is readout in a normal manner, whilst the next row is illuminated and the process repeated.","The numbers stored in each latch are proportional to the time of flight allowing the distance to each of the points illuminated by the laser line and imaged by the photodiodes to be measured.","To improve performance the time of flight measurement circuitry can use the same principles described above.","With this approach, parallax effects must be taken into account for objects close to the optical system.","These effects may be minimised either by configuring the laser source and receiving optics to be as close as possible, or preferably co-incident through the use of a beam splitter.","Alternatively, the laser line can be defocused to illuminate a broad strip of the remote object and ensure that a single row of photodiodes will all receive reflected light.","Another approach is for the control electronics to cause not one, but several photodiodes in each column to be connected to the column amplifier at once.","These techniques can be used singly or in combination.","To remove the need for any moving parts, an illumination system comprising multiple pulsed or modulated light sources can be used.","This approach is illustrated in FIG.","6 which shows an illumination system comprising three laser diodes 90A, 90B, 90C configured so that each laser diode illuminates a region 91A, 91B, 91C of the field of view 92 of the receiving optics.","Each light source would be operated in sequence to cause the region of illumination to be synchronised with the readout of the Y scanned, X parallel output array.","Preferably, the light sources; e.g.","laser diodes or light emitting diodes (LED) are fabricated on a common substrate with integrated drive circuitry to minimise cost.","To steer and focus the output of the each individual light source, it is advantageous to fabricate an array of micro-lenses, each aligned with an individual light source.","It can be seen that many alternative laser/multiplexer combinations can be used, depending upon the requirements of the end application.","For example, if it is only required to measure the distance profile along a line, then a one dimensional array of photodiodes may be used.","To those skilled in the art of imaging, it will be seen that it is also possible to use a charge coupled device (CCD) as the detector array for measuring the distance profile along a line.","This is particularly appropriate with modulated light sources.","In this case, a two dimensional CCD array is masked so that the top row of image section pixels, furthest from the parallel readout register is illuminated.","The CCD image section is then clocked in a time delay and integrate (TDI) mode in synchronism with the light source modulation to capture the reflected modulation sequence for each pixel in the illuminated row in parallel.","The captured sequence can then be readout of the CCD in the normal manner and the data from each column processed using cross correlation to extract the time of flight from the light source to the illuminated pixel at the top of each column.","Alternatively, a very simple system can be realised using a single bright pulsed or modulated light source to illuminate the whole of the field of view simultaneously, and an X-Y addressed array, or a Y scanned, X parallel output array used to capture the 3D image data in the manner described above.","This enables an imaging system to be made with no moving parts.","The light source is operated once for each group of pixels of the detector array to be read out simultaneously.","For the very highest speed applications, TOF measurement circuitry can be provided for each photodiode in the array.","Thus, the group of pixels is the entire array of pixels.","This enables the TOF and hence distance to the remote object to be measured in parallel for all points in the field of view.","However, in a lower cost implementation, different regions of the light detector are actuated separately, in a sequence.","The sequence may be row-by-row or column-by-column, depending on the row and column driver circuit configurations.","This enables different sub-regions (rows or columns) of the light detector to be actuated in sequence, with a different operation of the light source being allocated to each sub-region.","In all examples above, the distance measurement mode of operation of the detector pixel array involves a direct-read mode of operation, in which photogenerated charges pass directly to the measurement circuitry, so that the time of arrival of incident light is known.","The imaging mode is an integrating mode of operation, in which charges are allowed to build up on the photodiode self-capacitance (or else on an addition charge storage capacitor in the pixel layout).","The row and column driver circuits will need to be operable in these two different modes, and this will typically require different control voltages and timing in the two modes.","The possible specific pixel configurations and control schemes will be apparent to those skilled in the art."]],"string":"[\n [\n \"Surface profile measurement\",\n \"An imaging system scans a light source output over an object to be imaged, and receives the reflected signal in a multi-region light detector, wherein different regions of the object to be imaged are imaged onto different regions of the detector by the receiving optics, and wherein different regions of the light detector can be actuated separately.\",\n \"The light source scanning is synchronised with the actuation of the light detector so that a region of the detector imaging the region of object being illuminated by the light source is actuated.\",\n \"The time of flight of light signals from the light source to the actuated portion of the detector is calculated for all scanning directions, to enable a three dimensional image of the object to be constructed.\"\n ],\n [\n \"1.An imaging system comprising: a light source; means for scanning the light from the light source over an object to be imaged; stationary receiving optics for receiving light reflected from the object to be imaged; a multi-region light detector for detecting light received from the receiving optics, wherein different regions of the object to be imaged are imaged onto different regions of the detector by the receiving optics, and wherein different regions of the light detector can be actuated separately; control means to synchronise the light source scanning and the actuation of the light detector so that a region of the detector imaging the region of the object being illuminated by the light source is actuated; and processing means for measuring the time of flight of light signals from the light source to the actuated portion of the detector for all scanning directions.\",\n \"2.An imaging system as claimed in claim 1, wherein the or each light source is a pulsed laser or light emitting diode or diodes.\",\n \"3.An imaging system as claimed in claim 1, wherein the light source is a laser or light emitting diode or diodes having a modulated output.\",\n \"4.An imaging system as claimed in any preceding claim, wherein the means for scanning comprises: a reflector for directing the light from the light source to an object to be imaged; and a drive mechanism for controlling movement of the reflector for scanning the light from the light source over the object to be imaged.\",\n \"5.An imaging system comprising: a plurality of light sources, each light source being directed to an object to be imaged; stationary receiving optics for receiving light reflected from the object to be imaged; a multi-region light detector for detecting light received from the receiving optics, wherein each light source is imaged by the receiving optics onto a different region of the light detector, wherein the different regions of the light detector can be actuated separately; control means to synchronise the operation of individual light sources and the operation of the light detector so that the region of the detector imaging the region of the object being illuminated by an operating light source is actuated; and processing means for measuring the time of flight of light signals from the operating light source to the actuated portion of the detector for all scanning directions.\",\n \"6.An imaging system as claimed in claim 5, wherein each light source comprises a pulsed laser or light emitting diode or diodes.\",\n \"7.An imaging system as claimed in claim 5, wherein the light source is a laser or light emitting diode or diodes having a modulated output.\",\n \"8.An imaging system comprising: a light source directed to an object to be imaged; stationary receiving optics for receiving light reflected from the object to be imaged; a multi-region light detector for detecting light received from the receiving optics; control means to synchronise the operation of the light source and the operation of the light detector so that a region of the detector is actuated synchronously with an allocated operation of the light source; and processing means for measuring the time of flight of light signals from the light source to the actuated portion of the detector.\",\n \"9.An imaging system as claimed in claim 8, wherein all regions of the light detector are actuated in parallel.\",\n \"10.An imaging system as claimed in claim 8, wherein different regions of the light detector can be actuated separately.\",\n \"11.An imaging system as claimed in claim 10, wherein different sub-regions of the light detector are actuated in sequence, a different operation of the light source being allocated to each sub-region.\",\n \"12.An imaging system as claimed in claim 10 or 11, wherein each sub region comprises a row or column of detector pixels of a pixel array.\",\n \"13.An imaging system as claimed in any one of claims 8 to 12, wherein the light source comprises a pulsed laser or light emitting diode or diodes.\",\n \"14.An imaging system as claimed in any one of claims 8 to 12, wherein the light source is a laser or light emitting diode or diodes having a modulated output.\",\n \"15.An imaging system as claimed in any preceding claim, wherein the light detector comprises a photodiode array.\",\n \"16.An imaging system as claimed in claim 15, wherein the photodiode array is operable in two modes, a first mode in which light signals from all photodiodes in the array are read out in sequence, and a second mode in which light signals from selected photodiodes or sub-groups of photodiodes in the array are read out in a sequence.\",\n \"17.An imaging system as claimed in claim 15 or 16, wherein the processing means is integrated onto the same substrate as the photodiode array.\",\n \"18.An imaging system as claimed in any one of claims 15 to 17, wherein multiple channels of time of flight measurement circuitry are integrated onto the same substrate as the photodiode array and configured to process signals from different actuated portions of the photodiode array in parallel.\",\n \"19.An imaging system as claimed in any preceding claim, wherein the processing means uses cross-correlation for measuring the time of flight of light signals from the light source to the actuated portion of the detector.\",\n \"20.A method of obtaining an image of an object comprising: scanning a light source signal over the object by directing a light source output in a plurality of scanning directions in sequence, and detecting reflected light received from the object using a two dimensional light detector array; determining the regions of the light detector array which are illuminated for each scanning direction; scanning the light source signal over the object again, and detecting reflected light received from the object using only the determined regions of the light detector array; calculating the time of flight of light pulses from the light source to the detector for each scanning direction; and obtaining a 3D profile from the time of flight calculations.\",\n \"21.A method as claimed in claim 20, wherein the scanning of the light source is synchronised with the actuation of selected regions of the light detector array.\",\n \"22.A method as claimed in claim 20 or 21 wherein the light source signal comprises a pulsed laser output or output of a light emitting diode or diodes.\",\n \"23.A method as claimed in claim 20 or 21, wherein the light source signal comprises a modulated output of a laser or light emitting diode or diodes.\",\n \"24.A method as claimed in claim 23, wherein calculating the time of flight uses cross-correlation.\"\n ],\n [\n \"The invention relates to apparatus for measuring the surface profile of a sample, and in particular relates to non-contact 3D surface profile measurement systems.\",\n \"The non-contact measurement of three-dimensional (3D) objects to extract data regarding their physical form and location in space is a topic which has excited much research.\",\n \"Many techniques have been developed to suit the distance to the object, the precision with which the object's features need to be measured and so on.\",\n \"One common technique is to illuminate the remote object with a light source of known pattern; so called “structured illumination”.\",\n \"A camera is located some distance away from the structured light source and arranged to collect an image of the projection of the structured light pattern onto the surface of the remote object.\",\n \"FIG.\",\n \"1 shows this known profile measurement using structured lighting.\",\n \"A structured light source 10 projects light onto the object 12, and the reflected light is captured by a camera 14 having a field of view 15 which covers the object 12.An image processor 16 derives the 3D profile data 18 by analysis of the deformation of the structured light pattern, which is representative of the distance to the remote object.\",\n \"The 3D nature of the remote object causes the structured light pattern in the image captured by the camera to be deformed.\",\n \"From knowledge of the physical separation of the camera from the light source (the baseline 20) and trigonometry, it is possible to compute the 3D surface profile of the remote object from the deformation of the image of the structured light pattern.\",\n \"A benefit of this system is that if a video camera is used to capture the image, then with sufficient processing, the 3D surface profile can be measured at video frame rates (50 or 60 times per second).\",\n \"As this system works on trigonometric principles, the depth accuracy is related to the baseline of the system and the resolution of the sensor.\",\n \"As a result, practical considerations tend to limit the application of such systems to remote objects no more than a few metres away.\",\n \"In addition, as the baseline is lengthened, the angle between the incident light pattern and the line of sight of the camera becomes more obtuse and shadows cast by features in the remote object can obscure the 3D profile measurement.\",\n \"Alternative techniques rely on scanning a device which measures the distance to a remote point; e.g.\",\n \"a laser rangefinder (LRF) or interferometer, using a remote controlled pan and tilt unit.\",\n \"An example of such a system is illustrated in FIG.\",\n \"2.A distance measuring device such as a laser rangefinder 30 measures the distance to a single point on the object, and is controlled by a pan-and-tilt unit 32.Control electronics 34 under the control of a computer causes the pan and tilt unit 32 to scan the line of sight of the distance measuring device 30 across the object to be measured to build up a 3D matrix of the azimuth, elevation and distance to the remote object.\",\n \"This matrix of numbers represents the 3D surface profile of the remote object measured in polar co-ordinates from the axis of rotation of the pan and tilt unit.\",\n \"Some corrections may be applied to the data if the axes of rotation of the pan and tilt unit are not co-incident with the line of sight of the measuring device.\",\n \"The resultant 3D surface profile 18 can be transformed to other co-ordinate systems using known mathematical techniques.\",\n \"This technique is analogous to radar systems where the time of flight of a scanned micro-wave radio signal is used to determine the distance to one or more remote objects.\",\n \"This scanned LRF approach does not suffer from the range limitations of the structured light approach, but is relatively expensive to implement because high precision and costly components are required for good accuracy.\",\n \"In addition, because the whole mass of the distance measuring device is scanned, achieving scanning at rates sufficient to give “real time” 3D image data is problematic.\",\n \"To overcome this difficulty, systems have been built using a scanned mirror or mirrors in front of the LRF.\",\n \"For example PCT/US97/06793 describes a complex system using two orthogonal mirrors to scan the LRF beam horizontally and vertically so as to cover a field of view of approximately 40°×40°.\",\n \"This approach enables the LRF to remain static.\",\n \"However, the mirror/mirrors must be sufficiently large to encompass both the laser beam and lens aperture of the receiving system without vignetting or optical crosstalk between the emitted laser beam and received beam and so such mirror scanned systems still remain expensive.\",\n \"Hybrid approaches, where the distance measuring device is scanned along a line in one direction and the remote object is rotated have been used to create a map of the complete surface of a the remote object.\",\n \"However, these techniques are not appropriate for all targets.\",\n \"Other workers have sought to overcome these problems by scanning the laser beam whilst keeping the detector static, as described in GB 2 286 495.With this approach, the only moving part of the system is a scanned mirror which only need be sufficiently large to steer the laser beam.\",\n \"This avoids the high cost of a precision motorised pan and tilt head and enables a high scan rate.\",\n \"Furthermore, because the laser and receiving optical paths can be kept completely separate there is no risk of optical crosstalk.\",\n \"However, this approach requires the field of view of the receiving optical system to encompass the whole of the remote object.\",\n \"As a result, the optical system will collect illumination from the whole of the field of view.\",\n \"This generates a background offset signal level in the photodetector of the optical system.\",\n \"Whilst this offset level can be compensated for in the signal detection process, the shot noise inherent in the offset signal level significantly degrades the ratio of detected laser signal level to noise and hence reduces the maximum range of the system.\",\n \"To overcome this significant disadvantage some very complex schemes have been proposed.\",\n \"One such is as described in U.S. Pat.\",\n \"No.\",\n \"5,682,229 where the reflected light from a scanning laser is focussed onto a stationary detector assembly which uses a series of complex fibre bundles to transfer the collected light to 128 separate avalanche photodiodes and associated time of flight measuring circuitry.\",\n \"It will be appreciated that the complexity and use of relatively expensive components such as coherent fibre-optic assemblies and multiple discrete avalanche photodiodes makes such approaches expensive to implement.\",\n \"According to the invention, there is provided an imaging system comprising: a light source; means for scanning the light from the light source over an object to be imaged; stationary receiving optics for receiving light reflected from the object to be imaged; a multi-region light detector for detecting light received from the receiving optics, wherein different regions of the object to be imaged are imaged onto different regions of the detector by the receiving optics and wherein different regions of the light detector can be actuated separately; control means to synchronise the light source scanning and the actuation of the light detector so that a region of the detector imaging the region of the object being illuminated by the light source is actuated; and processing means for measuring the time of flight of light signals from the light source to the actuated portion of the detector for all scanning directions.\",\n \"The direction of scanning and the associated measured time of flight can be used to build up a three dimensional image of the object.\",\n \"The arrangement of the invention provides a simple structure with few moving parts.\",\n \"The means for scanning may comprise a reflector for directing the light from the light source to an object to be imaged, and a drive mechanism for controlling movement of the reflector for scanning the light from the light source over the object to be imaged.\",\n \"According to a second aspect of the invention, there is provided an imaging system comprising: a light source directed to an object to be imaged; stationary receiving optics for receiving light reflected from the object to be imaged; a multi-region light detector for detecting light received from the receiving optics; control means to synchronise the operation of the light source and the operation of the light detector so that a region of the detector is actuated synchronously with an allocated operation of the light source; and processing means for measuring the time of flight of light signals from the light source to the actuated portion of the detector.\",\n \"This aspect enables an imaging system to be made with no moving parts, using a multi-region light detector.\",\n \"All regions of the light detector can be actuated in parallel, for high speed image capture.\",\n \"However, different regions of the light detector can preferably be actuated separately.\",\n \"This enables different sub-regions of the light detector to be actuated in sequence, a different operation of the light source being allocated to each sub-region.\",\n \"For example, each sub region may comprises a row or column of detector pixels of a pixel array.\",\n \"According to a third aspect of the invention, there is provided an imaging system comprising: a plurality of light sources, each light source being directed to an object to be imaged; stationary receiving optics for receiving light reflected from the object to be imaged; a multi-region light detector for detecting light received from the receiving optics, wherein each light source is imaged by the receiving optics onto a different region of the light detector, wherein the different regions of the light detector can be actuated separately; control means to synchronise the operation of individual light sources and the operation of the light detector so that the region of the detector imaging the region of the object being illuminated by an operating light source is actuated; and processing means for measuring the time of flight of light signals from the operating light source to the actuated portion of the detector for all scanning directions.\",\n \"According to this aspect, the need to scan the light source output over the object to be imaged is again avoided.\",\n \"In each of these aspects, the or each light source may comprise a pulsed laser or light emitting diode or diodes, and the output may be modulated, with cross correlation techniques used for the time of flight measurement.\",\n \"In each of these aspects of the invention, the light detector preferably comprises a photodiode array, which can be implemented at low cost.\",\n \"The photodiode array may be operable in two modes; a first mode in which light signals from all photodiodes in the array are read out in sequence, and a second mode in which light signals from selected photodiodes or sub-groups of photodiodes in the array are read out in a sequence.\",\n \"The invention also provides a method of obtaining an image of an object, comprising: scanning a light source signal over the object by directing a light source output in a plurality of scanning directions in sequence, and detecting reflected light received from the object using a two dimensional light detector array; determining the regions of the light detector array which are illuminated for each scanning direction; scanning the light source signal over the object again, and detecting reflected light received from the object using only the determined regions of the light detector array; calculating the time of flight of light pulses from the light source to the detector for each scanning direction; and obtaining a 3D profile from the time of flight calculations.\",\n \"Examples of the invention will now be described in detail with reference to the accompanying drawings, in which: FIG.\",\n \"1 shows a first known 3D imaging system; FIG.\",\n \"2 shows a second know 3D imaging system; FIG.\",\n \"3 shows a simplified example of the photodiode array for use in the system of the invention; FIG.\",\n \"4 shows an imaging system of the invention, using the photodiode array of FIG.\",\n \"3; FIG.\",\n \"5 shows a modification to the photodiode array; FIG.\",\n \"6 shows how pulsed light sources can be used in the imaging system of the invention.\",\n \"The invention provides a 3D imaging system, with a stationary receiving optical system.\",\n \"The invention uses a two dimensional detector, which will first be described with reference to FIG.\",\n \"3.FIG.\",\n \"3 is a simplified schematic of a photodiode array for use in an imaging system of the invention, in a 2×2 format.\",\n \"The device consists of an array of photodiode pixels 60, each of which comprises a photodiode (PD11 to PD22) and associated transistor (TR11 to TR22), which are configured and drive to act as analogue switches.\",\n \"For standard video imaging applications, the device is operated in an integration mode where incident illumination is focussed upon its surface.\",\n \"The incident illumination generates charge within each photodiode by the photoelectric effect.\",\n \"During this integration period, connections X1, X2, Y1 and Y2 are all held low so that all transistors are off and the photodiodes are electrically isolated.\",\n \"The photo-generated charge then accumulates in each photodiode and is stored on the self-capacitance of the photodiode.\",\n \"Once sufficient photocharge has been collected, the device is readout as follows.\",\n \"Input X1 is taken to a high potential so that TR1 is turned on thereby allowing charge to flow between the column and a charge sensitive amplifier 62.Then input Y1 is pulsed high for addressing a row of pixels, turning TR11 on allowing and the photo-generated charge stored on photodiode PD11 to flow through TR11 and TR1 to the output amplifier 62 where the charge is converted to a voltage.\",\n \"This creates an output signal whose amplitude is proportional to the level of charge stored on PD11 and hence the level of light incident on TR11.After the self capacitance of PD11 has been discharged, input Y1 is taken low and input Y2 is taken high, allowing the stored charge on PD12 to be readout.\",\n \"In this way, a column of pixels is read out in turn.\",\n \"After all the charge collected by PD12 has been discharged, Y2 is taken low and X2 is taken high to allow PD21 and PD22 (the pixels in the next column) to be readout sequentially by pulsing Y1 and Y2 in the manner described above.\",\n \"It can be seen that this process allows the 2×2 array to be scanned and an electrical signal that is the analogue of the incident illumination generated.\",\n \"In normal operation, larger numbers of photodiodes are used, e.g.\",\n \"256×256, to increase resolution.\",\n \"Often the readout sequence and sensor scanning are arranged to generate a standard video signal.\",\n \"In addition, it may be noted that the basic structure described here has been simplified for the purpose of describing the proposed invention.\",\n \"Practical X-Y addressed photodiode arrays are generally fabricated as single complementary metal oxide semiconductor (CMOS) large scale integrated circuits (LSI) which include many refinements such as on-chip clock circuitry to generate the pulse sequences for electrodes X1 to Xn and Y1 to Yn.\",\n \"On-chip and additional pixel and/or column level circuitry improve amplification and detection of the photo-charge.\",\n \"These known refinements can be applied with advantage to the 3D profile measurement principle described here.\",\n \"For 3D profile measurement, the inventor has realised that such X-Y addressed photo-diode arrays can be utilised not in an integrating mode, but as a multiplexer synchronised with the laser scanning and pulsing so that only the individual photodiode receiving the reflected image of the laser spot on the remote object is addressed.\",\n \"FIG.\",\n \"4 shows a system using the photodiode array in this synchronised mode of operation.\",\n \"A sequentially pulsed laser beam output from a laser 40 is scanned by known means to illuminate a point on the surface of the remote object 12.To minimise size and cost, the laser scanning system 44 may be implemented in a number of ways including using electromagnetically or piezo-electrically scanned mirrors 42 or by mounting a laser chip on a micro-machined silicon or compact piezo electric structure.\",\n \"An image of the laser spot is formed on the surface of the X-Y addressed photodiode array 70 by the stationary, receiving optical system 46.A narrow band-pass optical filter 50 may be used to reject all wavelengths except those near the laser wavelength and optimise discrimination of the laser pulse against the background illumination.\",\n \"The control electronics apply logic level signals to the relevant X and Y control lines of the X-Y addressed array so that the photodiode illuminated by the image of the laser spot is connected to the pre-amplifier and time of flight detection electronics 56.In particular, the reflected laser pulse is captured by this photodiode and the resultant electrical signal routed to an electrical pulse detector 54 and TOF (time of flight) measurement circuitry 56.This computes the TOF of the laser pulse to the spot on the remote object and back to the photodiode on the X-Y addressed array and hence distance from the remote object to the X-Y addressed array.\",\n \"This process is then repeated for many points over the surface of the remote object to measure the surface profile of the remote object.\",\n \"If the image formed of the laser spot is larger than a single pixel then the control electronics can cause the detector to address a group of adjacent photodiodes (e.g.\",\n \"a 2×2 s-array of photodiodes) in parallel to optimise collection and detection of the laser energy.\",\n \"It can be seen that because only one or a small number of the photodiodes is connected to the receiving amplifier and time of flight electronics at any one time, the background offset signal will be limited to that generated by the part of the field of view focussed onto the individual photodiode/photodiode group, rather than from the whole of the field of view of the optics as for the system described above.\",\n \"For example, if the laser beam is arranged to sequentially scan 100×100 equispaced points within the optical field of view, then the background signal level collected by each photodiode/photodiode group will be nominally reduced by 10,000 times in comparison to the simple system described above, which confers substantial benefits on system performance.\",\n \"It may be noted that a simplified version of this system can be used with a linear photodiode array to measure a 2D, rather than 3D surface profile.\",\n \"The performance of the system can be substantially improved by replacing the pulsed laser source 40 with a modulated laser source and the pulse discriminator 56 by a cross-correlation system.\",\n \"Such systems are known, for example, from DE19949803 to Denso Corp and PCT/GB/04968.In particular, the system may include a signal source such as a laser for supplying a modulation signal and a transmission system connected to the signal source for transmitting a transmitted optical signal modulated by the modulation signal.\",\n \"The modulation signal may be, for example a maximal length sequence.\",\n \"A reception system is then provided for receiving a reflected and delayed version of the transmitted signal, and a cross-correlator for obtaining the time delay.\",\n \"The cross correlator can be arranged to determine, at a coarse resolution, the time delay of the modulation signal needed to maximise the correlation between the time delayed modulation signal and the received signal.\",\n \"The cross correlator can then determine, at a finer resolution than the coarse resolution, the correlation between the modulation signal and the received signal as a function of the time delay of the modulation signal with respect to the received signal in a smaller time delay range around the determined time delay.\",\n \"A measure of distance is calculated from the time delay of the modulation signal needed to maximise the correlation between the time delayed modulation signal and the received signal.\",\n \"The cross-correlator can be implemented digitally.\",\n \"In this way, for a given laser peak power, greater energy can then be delivered to the remote object which improves the signal to noise ratio and hence maximum range of the system.\",\n \"This oversampling approach enables the distance resolution of the system to be improved, and the efficient signal processing method using coarse and fine cross-correlators minimises the processing power needed.\",\n \"In a preferred embodiment, the X-Y sensor, time of flight measurement system and control electronics are fabricated on a single integrated circuit to minimise manufacturing cost.\",\n \"The photodiodes can be manufactured and operated as avalanche photodiodes to provide signal amplification by the avalanche effect, prior to signal detection.\",\n \"The time of flight measurement approach used may be as described above in connection with FIG.\",\n \"4 or other higher performance schemes described above.\",\n \"X-Y Sensors such as described here can suffer from image lag which arises because a) the photo charge generated by incident light upon a pixel is partitioned between the photodiode's intrinsic capacitance and the capacitance of the row and column electrodes resulting in some charge being left on the row and column electrode capacitances after each pixel is read out and b) a load transistor may be provided to discharge the photodiode capacitance and bias the amplifier, but the effective resistance of this load transistor increases with declining current increasing the time constant of the system at low light levels.\",\n \"In a preferred embodiment, this effect may be minimised by configuring the receiving amplifier as a transimpedance amplifier whilst the photodiode array is used in a multiplexer mode.\",\n \"The virtual earth offered by the transimpedance amplifier's input provides a low impedance path through which the photo generated current can flow, reducing the charge partitioning effect and improving system bandwidth.\",\n \"This approach also provides a means to bias the photodiodes during readout.\",\n \"In an optimal development of the preferred embodiment, a transimpedance amplifier stage is provided for each pixel to drive the multiplexing circuitry of the array.\",\n \"The laser scanning pattern will often be a repeating pattern arranged to cover the optical field of view whilst providing adequate time resolution to measure the position of moving objects in the field of view.\",\n \"The pattern is typically arranged as a conventional raster scan for ease of display on conventional monitors.\",\n \"However, it can be seen that other patterns may be used.\",\n \"One useful pattern is a spiral scan pattern where by controlling the velocity of the laser scan, increased spatial resolution may be achieved in the centre of the scan whilst still maintaining a low spatial resolution to detect objects appearing at the periphery of the scan.\",\n \"For those applications where it is necessary to monitor the 3D motion of a few specific objects within the field of view, the scan can be controlled adaptively to track the objects and ignore the rest of the field of view.\",\n \"This approach can increase the temporal resolution for the tracked objects.\",\n \"An important benefit of the approach described here is that the X-Y addressed array can still be operated in an imaging mode, rather than a multiplexed time of flight detector mode.\",\n \"This can be achieved simply by returning the sequence of pulses applied to the X-Y addressed array to a conventional video scanning sequence.\",\n \"This has several significant benefits.\",\n \"First, for optimal performance, it is important that only the photodiode or local group of photodiodes receiving the image of the laser spot at any point in time are addressed; i.e.\",\n \"that the laser scanning and the photodiode array scanning are synchronised.\",\n \"This would normally require precise calibration of the scanner and optical system.\",\n \"However, if the laser is scanned whilst the sensor is in an imaging mode, an image of the laser path can be collected by the control electronics.\",\n \"This image can be used to determine the precise path of the laser beam image on the surface of the photodiode array and hence set up the correct addressing sequence for the X-Y addressed array and/or laser pulsing sequence, to ensure synchronisation in the multiplexed time of flight detector mode.\",\n \"Thus, the normal addressing mode is used as a calibration stage for the higher performance multiplexing mode.\",\n \"In effect, the system can be self calibrating which is a major benefit for systems which have to operate over large temperature ranges.\",\n \"Secondly, if the laser is turned off and the X-Y addressed photodiode array is operated in an imaging mode, then the system can be used as a conventional imaging device; i.e.\",\n \"a video or still camera to provide additional information regarding the remote object.\",\n \"Third, because the same detector is used to capture a 3D surface profile and standard image, the registration between the standard image and 3D data is near perfect.\",\n \"This enables beneficial combinations of conventional and imaging and 3D profile measuring modes to be used.\",\n \"For example, by toggling the system between capturing a 3D scan and a conventional image scan both a video image sequence and 3D sequence to be captured and overlaid on one another.\",\n \"This approach is particularly beneficial for collision avoidance or intruder detection.\",\n \"If the 3D image shows that an object within the field of view is too close or is on a path likely to lead to a collision, the corresponding part of the conventional image can be coloured or caused to flash to draw, attention to the problem.\",\n \"Another benefit of capturing perfectly registered 3D and conventional image data is that image processing methods can be applied simultaneously to both sets of data, using the optimal method to extract important features from each image and combining the feature data to yield more information about objects in the field of view.\",\n \"For example, the 3D data can be used to determine the orientation and size of an object in the field of view.\",\n \"This data can then be used to select, or generate from a 3D solid model of a known object, an appropriate pattern for pattern matching and object/target recognition.\",\n \"It can be seen that the benefits of acquiring perfectly registered 3D and conventional image data are many.\",\n \"Where higher scanning rates are required, the system described above can be improved by modifying the optics of the laser to illuminate a line, rather than a point, of the remote object and modifying the structure of the X-Y addressed array to permit a row of photodiodes to be connected in parallel to multiple time of flight measurement processors; i.e.\",\n \"a Y scanned, X parallel output array.\",\n \"One implementation of the revised photodiode array structure is shown for a 2×2 array in FIG.\",\n \"5.Each column switch transistor has been replaced by an amplifier 80 and time of flight measurement (TOF) circuitry comprising a pulse detector 82, counter 84 and latch 86.All counters 84 are clocked at the same frequency by a common clock input 88.The laser scanning system is set to illuminate a strip of the remote object.\",\n \"The image of the illuminated strip is focussed onto row N of the photodiode array.\",\n \"The laser is pulsed on, the TOF counters 84 are reset to zero and the control electronics takes the relevant Y electrode (Y1,Y2) high to connect all the photodiodes along the row to the corresponding amplifier & TOF circuitry.\",\n \"As each photodiode receives the reflected pulse of laser light, the pulse detector causes the corresponding counter 84 to stop counting.\",\n \"Once all reflected pulses have been received, the number held on each counter 84 is latched into the latches 86 and is readout in a normal manner, whilst the next row is illuminated and the process repeated.\",\n \"The numbers stored in each latch are proportional to the time of flight allowing the distance to each of the points illuminated by the laser line and imaged by the photodiodes to be measured.\",\n \"To improve performance the time of flight measurement circuitry can use the same principles described above.\",\n \"With this approach, parallax effects must be taken into account for objects close to the optical system.\",\n \"These effects may be minimised either by configuring the laser source and receiving optics to be as close as possible, or preferably co-incident through the use of a beam splitter.\",\n \"Alternatively, the laser line can be defocused to illuminate a broad strip of the remote object and ensure that a single row of photodiodes will all receive reflected light.\",\n \"Another approach is for the control electronics to cause not one, but several photodiodes in each column to be connected to the column amplifier at once.\",\n \"These techniques can be used singly or in combination.\",\n \"To remove the need for any moving parts, an illumination system comprising multiple pulsed or modulated light sources can be used.\",\n \"This approach is illustrated in FIG.\",\n \"6 which shows an illumination system comprising three laser diodes 90A, 90B, 90C configured so that each laser diode illuminates a region 91A, 91B, 91C of the field of view 92 of the receiving optics.\",\n \"Each light source would be operated in sequence to cause the region of illumination to be synchronised with the readout of the Y scanned, X parallel output array.\",\n \"Preferably, the light sources; e.g.\",\n \"laser diodes or light emitting diodes (LED) are fabricated on a common substrate with integrated drive circuitry to minimise cost.\",\n \"To steer and focus the output of the each individual light source, it is advantageous to fabricate an array of micro-lenses, each aligned with an individual light source.\",\n \"It can be seen that many alternative laser/multiplexer combinations can be used, depending upon the requirements of the end application.\",\n \"For example, if it is only required to measure the distance profile along a line, then a one dimensional array of photodiodes may be used.\",\n \"To those skilled in the art of imaging, it will be seen that it is also possible to use a charge coupled device (CCD) as the detector array for measuring the distance profile along a line.\",\n \"This is particularly appropriate with modulated light sources.\",\n \"In this case, a two dimensional CCD array is masked so that the top row of image section pixels, furthest from the parallel readout register is illuminated.\",\n \"The CCD image section is then clocked in a time delay and integrate (TDI) mode in synchronism with the light source modulation to capture the reflected modulation sequence for each pixel in the illuminated row in parallel.\",\n \"The captured sequence can then be readout of the CCD in the normal manner and the data from each column processed using cross correlation to extract the time of flight from the light source to the illuminated pixel at the top of each column.\",\n \"Alternatively, a very simple system can be realised using a single bright pulsed or modulated light source to illuminate the whole of the field of view simultaneously, and an X-Y addressed array, or a Y scanned, X parallel output array used to capture the 3D image data in the manner described above.\",\n \"This enables an imaging system to be made with no moving parts.\",\n \"The light source is operated once for each group of pixels of the detector array to be read out simultaneously.\",\n \"For the very highest speed applications, TOF measurement circuitry can be provided for each photodiode in the array.\",\n \"Thus, the group of pixels is the entire array of pixels.\",\n \"This enables the TOF and hence distance to the remote object to be measured in parallel for all points in the field of view.\",\n \"However, in a lower cost implementation, different regions of the light detector are actuated separately, in a sequence.\",\n \"The sequence may be row-by-row or column-by-column, depending on the row and column driver circuit configurations.\",\n \"This enables different sub-regions (rows or columns) of the light detector to be actuated in sequence, with a different operation of the light source being allocated to each sub-region.\",\n \"In all examples above, the distance measurement mode of operation of the detector pixel array involves a direct-read mode of operation, in which photogenerated charges pass directly to the measurement circuitry, so that the time of arrival of incident light is known.\",\n \"The imaging mode is an integrating mode of operation, in which charges are allowed to build up on the photodiode self-capacitance (or else on an addition charge storage capacitor in the pixel layout).\",\n \"The row and column driver circuits will need to be operable in these two different modes, and this will typically require different control voltages and timing in the two modes.\",\n \"The possible specific pixel configurations and control schemes will be apparent to those skilled in the art.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10474293"}}},{"rowIdx":795,"cells":{"text":{"kind":"list like","value":[["Supported double layer structure for displaying a nucleic acid associated with a protein","The invention concerns novel biosensors, in particular a support for displaying nucleic acids and for detecting both the presence of nucleic acids in a sample and the linkage between proteins and nucleic acids, as well as the linkage between a ligand and a protein linked to a nucleic acid."],["1.A cell for displaying a nucleic acid, comprising: a support which is substantially flat on an atomic scale, a protein III having an unequivocal three-dimensional structure, a nucleic acid IV linked to said protein III, the nucleic acid-protein assembly being laterally mobile relative to said support, such that the hybridization of a target nucleic acid to said nucleic acid IV or the change in conformation of said nucleic acid IV can be detected via the geometric reorganization of the nucleic acid-protein assembly.","2.The cell as claimed in claim 1, characterized in that said protein III is chosen from the group consisting of proteins having a redox center and proteins having an anisotropic optical absorption property.","3.The cell as claimed in one of claims 1 and 2, comprising: a support which is substantially flat on an atomic scale, to which is attached a hydrophobic monolayer I, on which is present a monolayer II comprising phospholipids, a bridging molecule having a hydrophobic end which can interact with the monolayer II, and an end which is chemically functionalized so as to form a stable bond with a protein III, a protein III having an unequivocal three-dimensional structure, said protein III being laterally mobile in said layer II, a nucleic acid IV unequivocally attached to said protein III, preferably via a molecular arm.","4.The cell as claimed in one of claims 1 to 3, characterized in that the nucleic acid is attached to said protein via a covalent bond.","5.The cell as claimed in one of claims 1 to 3, characterized in that the nucleic acid is attached to said protein via a reversible bond.","6.The cell as claimed in one of claims 1 to 3, characterized in that the nucleic acid IV is attached to said protein III via a compound chosen from: a spacer arm connecting two identical or different, chemically reactive residues of the nucleic acid IV and of the protein III, chosen from the group consisting of thiols, amines, amides and arginines; a metal complex such as cis-platinum, trans-platinum, europium, or a complex of nickel, of copper or of ruthenium coordinated, firstly, with an amino residue or a group of amino residues of said protein III and, secondly, with one or more natural or modified bases of the nucleic acid IV, a covalent or noncovalent protein complex involving the nucleic acid IV.","7.The cell as claimed in one of claims 1 to 6, characterized in that said protein III has undergone a modification which introduces a unique site for effecting said bond [lacuna] said nucleic acid IV in an unequivocal manner.","8.The cell as claimed in claim 7, characterized in that said modification consists of a mutation of said protein so as to leave remaining or to introduce just one unique predetermined amino acid.","9.The cell as claimed in claim 8, characterized in that said predetermined amino acid is a cysteine.","10.The cell as claimed in one of claims 3 to 9, characterized in that said protein is attached to a phospholipid of the layer II.","11.The cell as claimed in claim 10, characterized in that said bond is a covalent bond, for example via acylation, farnesylation, via a GPI (glycosyl-phosphatidylinositol) anchoring sequence or via the attachment of a phospholipid to a specific residue of the protein III by any artificial method.","12.The cell as claimed in claim 10 or 11, characterized in that said bond is a reversible bond, for example via an arm capable of linking a metal chelate on one side and an amino acid or a group of amino acids on the other.","13.The cell as claimed in one of claims 10 to 12, characterized in that said protein is linked to a modified phospholipid of the layer II, said phospholipid having a chelate-binding zone.","14.The cell as claimed in one of claims 3 to 9, characterized in that said protein has a hydrophobic tail which allows it to be anchored in the layer II.","15.The cell as claimed in one of claims 3 to 14, characterized in that the phospholipids of the layer II have hydrophobic tails chosen such that said layer is not in the crystalline state, but in the fluid state at the temperature for use.","16.The cell as claimed in claim 15, characterized in that said hydrophobic tails are greater than or equal to 14 carbon atoms in chain length, said chain possibly having unsaturations, such that said layer II is in the fluid state at the temperature for use.","17.The cell as claimed in one of claims 3 to 16, characterized in that said phospholipids of the layer II have polar heads, chosen from the group consisting of neutral polar heads carrying no charges, polar heads which are neutral overall but which carry opposite charges (zwitterionic polar heads), or polar heads carrying an overall negative charge.","18.The cell as claimed in one of claims 3 to 17, characterized in that said layer I comprises hydrophobic elements with a chain length of approximately 2 to 2.5 nm, for example of between 12 and 20 carbon atoms, preferably between 14 and 18 carbon atoms.","19.The cell as claimed in one of claims 3 to 18, characterized in that said layer I comprises phospholipids linked to said support via their polar heads.","20.The cell as claimed in one of claims 3 to 18, characterized in that said layer I comprises hydrophobic elements linked to said support via a covalent bond, for example an alkyl-thiol bond or an alkyl-siloxane bond.","21.The cell as claimed in one of claims 1 to 20, characterized in that the flatness of said support is such that the difference in height between two zones separated by less than 100 nm is less than or equal to 10 nm, preferably less than or equal to 3 nm, more preferably less than or equal to 2 nm.","22.The cell as claimed in one of claims 1 to 21, characterized in that said support consists of a material chosen from the group consisting of glass covered with a layer of gold, cleaved mica, silicon or any other monocrystalline material.","23.The cell as claimed in one of claims 1 to 22, characterized in that it comprises several identical or different proteins linked to identical or different nucleic acids.","24.A support for detecting nucleic acids, displaying a plurality of cells as claimed in one of claims 1 to 23.25.The support as claimed in claim 24, characterized in that it comprises a system for measuring current, impedance or potential.","26.The support as claimed in claim 24, characterized in that it comprises, between each cell, an integrated optical device capable of measuring the absorbance of a cell in two cross directions and of deducing therefrom its state of organization (anisotropy) or of disorganization (isotropy).","27.The support as claimed in claim 24, characterized in that it comprises a surface alternating conducting and reflecting zones and nonconducting and transparent zones.","28.A method for identifying the presence of a test nucleic acid in a sample, comprising the steps of: a) bringing said sample into contact with a cell as claimed in one of claims 1 to 23, under conditions which allow the hybridization of said test nucleic acid to a nucleic acid attached to a protein of said cell, b) detecting the hybridization of said test nucleic acid to said nucleic acid attached to said protein.","29.A method for identifying the binding of a protein to a nucleic acid, and/or its activity on the conformation of said nucleic acid, comprising the steps of: a) bringing said protein into contact with a nucleic acid attached to a protein, in a cell as claimed in one of claims 1 to 23, b) detecting the binding of said protein to said nucleic acid, and/or its activity on the conformation of said nucleic acid.","30.A method for identifying the binding of a ligand to a protein attached to a nucleic acid attached to a protein in a cell as claimed in one of claims 1 to 23, comprising the steps of: a) bringing said ligand into contact with said protein, b) detecting the binding of said ligand to said protein.","31.A method for identifying the binding of a compound to a nucleic acid, and/or its activity on the conformation of said nucleic acid, comprising the steps of: a) bringing said compound into contact with a nucleic acid attached to a protein, in a cell as claimed in one of claims 1 to 23, b) detecting the binding of said compound to said nucleic acid, and/or its activity on the conformation of said nucleic acid.","32.The method of identification as claimed in one of claims 28 to 31, characterized in that the detection is carried out via an optical (absorbance, fluorescence), electrical, electron-based, surface plasmon resonance, energy transfer, radiolabeling, diffraction (optical, electron, X-ray, neutron diffraction) or microscopy (direct optical or fluorescence microscopy, electron microscopy, near-field microscopy, atomic force microscopy) system."],["The present invention relates to novel biostructures which can in particular be used as biosensors, in particular a support for displaying nucleic acids and for detecting both the presence of nucleic acids in a sample and the interaction between proteins and said nucleic acids, and also the binding between a ligand and a protein linked to a nucleic acid.","The development of DNA biosensors, also referred to as DNA chips, has considerably increased over the last few years.","The impact of such biosensors has mainly been directed toward sequencing, gene expression mapping, diagnosis (detection, damage, mutation, etc.)","and analysis of DNA-ligand interactions (hybridization, etc.).","Most of the biosensors employed use techniques for immobilizing single-stranded nucleic acid, or ssDNA, molecules, the analysis being carried out by virtue of optical transduction, electrochemical or piezoacoustic systems.","The fact that biological membranes are highly important in physiological terms as a site for numerous chemical and metabolic reactions has led to a craze for developing lipid models on inorganic surfaces.","The design of such biocompatible and biofunctional structures on solid substrates has made it possible to develop models involving interdisciplinary fields which have brought about an expansion of many lines of scientific research and also practical applications.","The main applications have thus concerned: the production of matrices for immobilizing enzymes, cell receptors or hormones in nondenaturing environments, the design of interfaces for capturing cells in order to study the effects of pharmaceutical molecules on the outcome of the cell, the self-assembly of very resistant membranes for the development of biosensors.","The analytical methods applied to the analysis of DNA-DNA biomolecular interactions, such as they are represented at the heart of DNA chip devices, are essentially based on the attachment of a single-stranded DNA (ssDNA) within a delimited area on a substrate.","The main techniques for producing DNAs covalently attached to the substrate are the in situ synthesis of ssDNA on the support, the assembly of ssDNAs which have been chemically modified to allow grafting onto the support, or microdeposition onto an adsorbent matrix (Boncheva et al., 1999, Berney et al., 2000, Wittung-Stafshede P. et al., 2000).","Now, in the development of biosensors, the arrangement of biomolecules at the interfaces plays a crucial role.","In order to obtain highly sensitive surfaces, it is important to exhibit receptor molecules in such a way that the corresponding ligand can interact without steric hindrances.","However, in these various approaches, the means used to produce these DNA matrices generate artifacts (errors in the in situ syntheses or in functionalization of oligonucleotides) which cause nonspecific adsorptions; thus, some of the current studies relate to means for getting round these various limitations (Steel et al., 2000).","Another limitation of these systems is inherent to the method of displaying the ssDNAs.","The highly compacted nature of the molecules on the surface leads to the appearance of steric constraints which affect the hybridization efficiency of the chip.","While the immobilization of ssDNAs on a surface is to date the most commonly used alternative, it also constitutes that which restricts the geometry of the device, resulting in a complete loss of flexibility and of modularity of the assembly.","One of the characteristics of the present invention is to provide a novel method of displaying nucleic acids at the surface of the substrate.","The method of displaying the nucleic acids according to the invention is original in that it allows the nucleic acids to be mobile, and thus makes it possible to detect any change in the state of said nucleic acids by detecting a transition between two states exhibiting distinct organizational levels.","Thus, the subject of the present invention is a cell for displaying a nucleic acid, comprising: a support which is substantially flat on an atomic scale, a protein III having an unequivocal three-dimensional structure (stable and defined spatial organization), a nucleic acid IV linked to said protein III, the nucleic acid-protein assembly being laterally mobile relative to said support, such that the hybridization of a target nucleic acid to said nucleic acid IV or the change in conformation of said nucleic acid IV can be or is detected via the geometric reorganization of the nucleic acid-protein assembly.","In a preferred case of the present invention, said protein III is chosen from the group consisting of proteins having a redox center and proteins having an anisotropic optical absorption property.","The protein III will be described in greater detail later.","The term “cell” is intended to mean a laterally delimited subset of the surface, which is characterized by a structural homogeneity, and many identical or variant copies of which can be reproduced on the inorganic support.","A cell can in particular be used for detecting a specific type of nucleic acid (as will be seen later), different from the nucleic acid detected by using another cell.","It is understood that the present invention also relates to a support displaying a plurality of cells according to the invention.","In the present application, the term “floater” may be used to denote the assembly consisting of the protein which is mobile relative to the support, attached to the oligonucleotide.","The oligonucleotide linked to the floater may be referred to as “primer” in the present application.","The following abbreviations may also be used in the present application: DMPC: dimyristoylphosphatidylchloline DOGS: 1,2-dioleoyl-sn-glycero-3-[(N-(5-amino-1-carboxy-pentyl)iminodiacetic acid)succinyl] DOPC: dioleoyl phosphatidylcholine DPDPB: 1,4-di-[3′-(2′-pyridyldithio)propionamido]butane DPPC: dipalmitoyl phosphatidylcholine dsDNA: double-stranded DNA DTT: dithiothreitol Hb5(His)4: human cytochrome b5-histidine 4 fusion protein Hb5(His)4mut24: cysteine (position 24) mutant of Hb5(His)4 MIAC: metal ion affinity chromatography OG: octyl glucopyranoside OM: octadecyl mercaptan SPR: surface plasmon resonance RU: response unit SAM: self assemblage monolayer ssDNA: single-stranded DNA It is advantageous for the oligonucleotide not to be directly in contact with the support, thereby allowing an improved display with respect to the target nucleic acids or other elements which interact and modify the conformation of the primer.","In a particular case, the protein-nucleic acid assembly is mobile relative to the support due to the fact that the protein is itself mobile in a lipid layer which is itself attached to the support.","A lipid bilayer is preferably used.","Briefly, this assembly consists of a lipid membrane reconstituted on a flat surface which is inorganic in nature.","This is then referred to as a supported lipid membrane.","Such a model has properties which are close to biological membranes in terms of compactness of the elements making it up, of compartmentalization, and of fluidity of the elements making up these models and also of most of the molecules incorporated or interacting with this model (Heyse et al., 1998, Marchal et al., 1998).","This property of membrane dynamics is the reason for the lateral mobility of the proposed structures and for the reorganization allowing signal transduction and domain formation.","In addition, it is possible to modulate as desired the molecular composition of this lipid structure in order to modify its intrinsic properties or to introduce new potentialities.","Thus, in a preferred embodiment, the present invention relates to a cell for displaying a nucleic acid, comprising: a support which is substantially flat on an atomic scale, to which is attached a hydrophobic monolayer I, on which is present a monolayer II comprising phospholipids, a bridging molecule having a hydrophobic end which can interact with the monolayer II, and an end which is chemically functionalized so as to form a stable bond with a protein III, a protein III having an unequivocal three-dimensional structure, said protein III being laterally mobile in said layer II, a nucleic acid IV unequivocally attached to said protein III, preferably via a molecular arm.","The fact that the protein is mobile in the layer II (lipid sheet) ensures lateral dynamics, i.e.","the possibility of the floater+primer assembly diffusing freely in two dimensions in the absence of any additional geometric constraint which would be imposed by interaction of the primer with other nucleic acids.","The membrane fluidity allows the reorganization and the contact of molecules essential for the self-assembly into superstructures.","In addition, in some embodiments, the supramolecular assembly thus produced is characterized by a strong modularity: each step for producing or for destructuring this assembly may be carried out irreversibly by the controlled action of the solutes present in an aqueous phase or by the action of thermal conditions or of an electrostatic environment, etc.","The grafting of a single-stranded nucleic acid onto the protein has been carried out so as to use the very particular properties of these biostructures.","In this way, the oligonucleotides are grafted onto blocks which are protein in nature and which are floating on the upper lipid monolayer of the membrane model.","This arrangement offers the following advantages: Lateral mobility of the floating protein anchor and, consequently, of the grafted ssDNA.","Suppression of the nonspecific interactions of the ssDNAs on the flat support due to an appropriate lipid covering.","Control of the density of the ssDNAs present at the surface of the biosensor (great modularity).","In some embodiments, reversibility of the various supramolecular interactions via aqueous treatment procedures (allowing regeneration of the biosensor).","The terms “nucleic acid”, “nucleic acid sequence”, “polynucleotide”, “oligonucleotide”, “polynucleotide sequence” or “nucleotide sequence”, terms which may be used indifferently in the present description, are intended to denote a precise series of natural or synthetic nucleotides defining a fragment or a region of a nucleic acid and which can correspond equally to a single-stranded, double-stranded or triplex DNA or RNA.","Thus, the nucleic acid sequences according to the invention also encompass PNAs (peptide nucleic acids), or the like.","The bonds between the various bases may be conventional phosphodiester bonds, or modified bonds such as phosphorothioates or methylphosphonates.","The oligonucleotides may also be chimeric, i.e.","they may have different bonds and/or bases.","The term thus encompasses any variant of a nucleic acid sequence which exhibits a property of recognition by hybridization.","Preferably, the nucleic acid linked to the protein in the cell according to the invention is a single-stranded nucleic acid.","Its size is not a determinant parameter according to the invention, and it can be readily chosen using the general knowledge of those skilled in the art.","In a preferred embodiment, the nucleic acid is attached to said protein via a reversible bond.","This reversible bond may, moreover, be covalent.","Thus, it is possible to attach the nucleic acid to the protein such that disulfide bonds are involved, which bonds can, however, be reversed in the presence of appropriate agents, or under particular conditions.","Thus, the reversible bond can, in this particular case, be considered to be a bond which can be readily broken.","In one embodiment, the nucleic acid is attached to said protein via a covalent bond.","Such a bond is known to those skilled in the art and can in particular be produced using modified bases in the oligonucleotide allowing coupling to a chemical arm which is functionalized so that it can be coupled specifically to the protein of the floater.","Moreover, there can be two types of grafting: a closely related grafting can be carried out so as to place the nucleic acid molecule and the protein tightly next to one another, or grafting via a molecular spacer arm can be carried out so as to provide greater conformational freedom of the DNA with respect to the floater.","A spacer arm attached in an unequivocal manner to said protein is therefore preferably used.","In fact, as will be developed later, it is advantageous for the geometry of the floater to be completely defined.","Thus, it is preferable for the spacer arm to be attached to the protein at a precise site of said protein.","In the embodiment in which the nucleic acid and the protein are placed next to one another, said spacer arm can then be a metal complex such as cis-platinum, trans-platinum, europium or another transition metal involving ligands originating from the very structure of the nucleic acid and one or more ligands originating from the very structure of the protein III of the floater.","When the spacer arm is longer, allowing greater flexibility of the nucleic acid, a spacer arm is preferably chosen which attaches a thiol- or amino-function introduced onto the nucleic acid, in particular via a modified base (it being possible for the base chosen to be any base, i.e.","it is possible to choose a base in the 3′ position, in the 5′ position or within the oligonucleotide, this alternative possibly having certain advantages, in particular of providing the oligonucleotide with a greater degree of freedom), to another functionality present, preferably uniquely present, on the protein.","This second function is preferably also a thiol function, carried for example by a cysteine.","However, other types of grafting of a nucleic acid to a protein exist.","They can in particular be grafted onto an arginine in the protein.","It is, however, preferable, in order to respect a principle of the invention, for the coupling of the nucleic acid to the protein to be specific, completely defined and unequivocal in molecular terms.","Thus, any type of random grafting of the nucleic acid onto the protein which may create an ambiguity or a heterogeneity in the structure of the floaters according to the invention is generally excluded.","Thus, preferably, the nucleic acid IV is attached to said protein III via a compound chosen from: a spacer arm connecting two identical or different, chemically reactive residues of the nucleic acid IV and of the protein III, chosen from the group consisting of thiols, amines, amides and arginines; a metal complex such as cis-platinum, trans-platinum, europium, or a complex of nickel, of copper or of ruthenium coordinated, firstly, with an amino residue or a group of amino residues of said protein III and, secondly, with one or more natural or modified bases of the nucleic acid IV, a covalent or noncovalent protein complex involving the nucleic acid IV.","The protein in the cell according to the present invention has a geometry which can be detected in an unequivocal manner, i.e.","a stable and defined spatial organization.","This means in particular that its conformation in the cell according to the present invention is identical whatever the protein observed, when the cell according to the invention displays several identical floaters.","This also means that bringing the cell into contact with outside nucleic acids capable of interacting with the primers in an unequivocal manner will lead to a highly repetitive organization of the floaters, in spatial terms, within the cell.","In particular, the external nucleic acids may bring about the association of floaters in organized groups and these groups of floaters may, by virtue of their lateral mobility, interact with one another noncovalently so as to form, within each cell, a network of floaters which is highly organized in two-dimensional terms.","The properties (physical, electrical, optical, quantum properties) of this highly organized network will then differ from the dynamic properties of disorganized floaters within the cell in the absence of any interaction with the external nucleic acid.","Preferably, the protein of the floater will be chosen such that the variations in spatial organization of the floaters relative to one another lead to a signal which can be detected by any method, in particular an electrical, optical or resonance method.","Proteins having this property will preferably be chosen from the group consisting of proteins having a redox center and/or an anisotropic optical absorption property and/or an intrinsic fluorescence property.","Mention is in particular made of cytochromes, and more preferably human cytochrome b5, yeast cytochrome b5 or cytochrome b5 of any other origin, flavodoxins, ferridoxins, azurins and other blue copper proteins, and multiheme proteins of the bacterial cytochrome c class, which have both a redox center and anisotropic optical absorption properties.","Among the fluorescent proteins, mention will preferably be made of green fluorescent protein (GFP) and analogs thereof obtained by site-directed mutagenesis or directed evolution.","In a preferred case of implementation, said protein is water soluble.","It may also have undergone modifications, some of which are indicated below, provided that these modifications result in a protein with a stable and defined three-dimensional structure satisfying the criteria as defined according to the present invention.","Thus, in a preferred embodiment, said protein has undergone a modification which has allowed the introduction of a unique site into said protein, in order to effect said bond with said nucleic acid.","The fact that a unique site is introduced into the protein according to the invention, in order to graft the nucleic acid, thus makes it possible to strongly promote the unicity, the specificity and the unequivocal definition of the protein-nucleic acid bond.","In a particular case, said modification consists of a mutation of said protein so as to leave remaining or to introduce just one predetermined unique amino acid.","The mutation can be introduced by mutation of the nucleic acid encoding the protein; this is the simplest method which will be favored by those skilled in the art.","The unique site for grafting the nucleic acid will then be this predetermined unique amino acid.","It is in fact within the scope of those skilled in the art to determine, according to the sequence of the protein which has been chosen in order to implement the invention, the amino acids to be modified so as to leave remaining just one of them, or so as to introduce one which is not present in the native protein.","Thus, and in order to illustrate this fact, when the protein which is used is a cytochrome b5, it is possible to introduce a cysteine into the protein, since this amino acid is absent from the native protein.","It is, moreover, always very advantageous to modify the protein such that it has only one cysteine, or at least such that it has a unique free cysteine (not involved in a disulfide bond with another cysteine).","It is also preferable, for correct implementation of the method according to the invention, for the modification in the protein to be such that this facilitates the grafting of the nucleic acid, i.e.","such that the amino acid used for said grafting is located at the periphery (at the surface) of said protein.","It is within the scope of those skilled in the art to determine, as a function of the protein chosen, the best position for introducing the modification into the protein.","In cytochrome b5, introduction of the cysteine by mutation of serine 24 has thus been exemplified.","However, alternative positions could easily have been chosen.","Thus, according to the three-dimensional structure of a cytochrome b5 of a mammal other than human (for example rat or bovine), it is entirely possible to select the amino acids to be modified so as to introduce just one cysteine into the sequence of the protein.","For example, the representation in three dimensions will make it possible to target the most peripheral amino acids of the protein, i.e.","those which promote better reactivity with the modified nucleic acid.","In addition, it may be of crucial advantage to link the ssDNA in the region of the redox center of the protein: a mutation of glycine 47 would make it possible to obtain such a connection.","One of the important characteristics of the present invention is that the protein III is mobile relative to the layer II of the cell according to the invention.","This also ensures the mobility of the nucleic acid IV attached to the protein III in the cell, relative to the support according to the invention.","The mobility of the protein in the layer II can be obtained in various ways, in particular when said protein has a hydrophobic tail which allows it to be anchored in the layer II.","The sequences of hydrophobic tails are known in the literature, and it is within the scope of those skilled in the art to choose such a sequence and to modify the selected protein by genetic engineering in order to introduce such an anchoring sequence.","It is then advantageous to choose a hydrophobic tail which is short, so that it becomes anchored only in the layer II, without penetrating into the layer I.","This makes it possible in particular to improve the mobility of the protein in the layer II, by reducing the “rubbing” phenomena which may be observed if the hydrophobic tail is too large.","The optimization of these elements remains within the scope of those skilled in the art.","In a preferred case, however, said protein is attached to a phospholipid of the layer II and, in a particular case, to an artificial phospholipid preferably introduced in a small proportion (i.e.","in a proportion less than 10%, preferably less than 5%, more preferably approximately 2%).","As a result, the mobility of the protein relative to the layer II is then similar to the mobility of the phospholipids in said layer.","Such a mobility is generally greater than when the protein has a hydrophobic tail.","This makes it possible to improve the levels of performance of the cell according to the invention, as will be seen.","Those skilled in the art have several methods available to them for producing a protein-phospholipid bond.","Mention may in particular be made, among covalent bonds, of acylation, farnesylation, the use of a GPI (glycosylphosphatidylinositol) anchoring sequence or the incorporation of a phospholipid or of a fatty acid into a specific residue of the protein III by any artificial method.","It may, however, be preferable to have a reversible bond, i.e.","a bond which can be broken in the presence of reagents or under particular conditions, but which is stable under the usual working conditions or in the presence of the usual working reagents (hybridization of nucleic acids, study of binding between biological elements, etc.).","In general, those skilled in the art can use all the known methods for attaching a protein to a liposome, for example, since liposomes generally consist of phospholipids.","Thus, it is in particular possible to use an arm capable of linking a metal chelate on one side and an amino acid on the other, using, in the layer II, a modified phospholipid, said phospholipid having a chelate-binding zone.","Thus, a preferred phospholipid according to the invention is DOGS, or 1,2-dioleoyl-sn-glycero-3-[(N-(5-amino-1-carboxypentyl)iminodiacetic acid)succinyl].","This phospholipid is a synthetic phospholipid possessing an iminodiacetate residue which complexes a nickel divalent cation.","In order to promote the mobility of the protein in the layer II, it is advantageous to choose the phospholipids of said layer II so that they have hydrophobic tails chosen such that said layer is not in the crystalline state, but in the fluid state at the temperature for use.","These properties of fluidity of a phospholipid layer are known to those skilled in the art, and there are in particular diagrams for choosing the chain length and the unsaturation in order to obtain fluidity at the temperature for use (Gulik-Krzywicki et al., 1967).","This temperature for use obviously depends on the use for which the sensor is intended.","Thus, when hybridization is sought, the temperature for use may be different from that used when the binding of a protein to a nucleic acid is studied.","Furthermore, transition between fluid and gel states may be strategic in the context of a detection technique based on super-structure imaging, such as atomic force microscopy (AFM).","Those skilled in the art will seek the phospholipids most suitable for the layer II as a function of the temperature for use.","As a general rule, and in order to simplify and improve the potentialities of the biosensor, fluidity of the layer II at ambient temperature is sought.","Thus, phospholipids having hydrophobic tails which are greater than or equal to 14 carbon atoms in chain length are preferably chosen, said chain optionally having unsaturations, such that said layer II is in the fluid state at the temperature for use.","It is advantageous to note, for example, that C14 elements are generally fluid at a temperature of greater than 23° C., whereas C16 elements are fluid only from 37° C., and C18 elements above 50-55° C. However, the use of C18 tails which have unsaturations makes it possible to decrease the phase transition (crystalline state-fluid state) temperature.","Nevertheless, for some applications or methods of detection, it may be preferred for the layer II not to be fluid at ambient temperature, but only at higher temperatures.","In particular, this method will be preferred if it is desired to “memorize” the organizational state of the floaters, using a decrease in temperature to fix the structure after an organization phase at higher temperature.","This will in particular be the case if there is a risk of the detection technique (near-field microscopy for example) or the conditions for analysis (dehydration for example) altering the structure.","The nonmodified phospholipids of the layer II have no particular characteristics besides the mobility properties already described; however, it is preferable for them to have polar heads, chosen from the group consisting of neutral polar heads carrying no charges, polar heads which are neutral overall but which carry opposite charges (zwitterionic polar heads), or polar heads carrying an overall negative charge.","The choice of the charge carried by the phospholipids may prove to be advantageous, in particular for reducing the phenomena of nonspecific absorption of the test nucleic acids on the layer, which are decreased when the charge of the phospholipid heads is negative.","The use of neutral heads (either uncharged or carrying several opposite charges) is also envisioned.","The layer II is assembled on the layer I conventionally, via the interactions which exist between two hydrophobic elements in an aqueous medium.","Reference may therefore be made to a self-assemblage layer.","However, in certain cases, a suitable catalyst, called fusogenic agent, will be used to facilitate this fusion.","Thus, such agents, such as Ca++ cations, the ethylene glycol polymer (PEG) or detergents, will be used according to protocols known to those skilled in the art.","As regards the layer I, it comprises hydrophobic elements with a chain length of approximately 2 to 2.5 nm, for example of between 12 and 20 carbon atoms, preferably between 14 and 18 carbon atoms.","It is in fact advisable to have a layer I which is particularly homogeneous.","Moreover, the chain length for the elements of the layer I is preferably sufficiently long so that the coming together of the two layers I and II forms a true bilayer.","The elements of the layer I are linear or branched and optionally carry unsaturations.","It is important for this layer I to be homogeneous, since it serves as a support for depositing the layer II.","The layer I is attached to the support of the cell according to the invention covalently, or via strong noncovalent bonds.","This method of bonding to the support is not of great importance for the implementation of the invention.","Thus, in a particular embodiment, said layer I comprises phospholipids linked to said support via their polar heads.","In another embodiment, said layer I comprises hydrophobic elements linked to said support via a covalent bond, for example an alkyl-thiol bond or an alkyl-siloxane bond, depending on the nature of the support.","It may be advantageous to have a support-layer I bond which is covalent, for facilitated re-use of the cell according to the invention.","In fact, when the nucleic acid is linked to the protein and the protein is linked to the layer II by reversible bonds, the cell can be readily regenerated by the action of elements (or conditions) which make it possible to break the nucleic acid-protein and protein-layer II bonds and to eliminate the layer II by the action of an appropriate detergent.","It is thus readily seen that a nonreversible covalent bond between the layer I and the support makes it possible to reconstruct a cell.","It is obvious that if the layer I is linked to the support via a reversible bond, it is possible to recommence preparing the cell, but with an additional step.","The re-use of the support can prove to be very advantageous insofar as said support must be virtually flat at the atomic level and, moreover, can include all or part of the system for detecting the state of the cells, which can pose certain manufacturing restrictions and increase the production costs for the cell according to the invention.","Thus, in a preferred embodiment, the flatness of said support is such that the difference in height between two zones less than 100 nm apart is less than or equal to 10 nm, preferably less than or equal to 3 nm, more preferably less than or equal to 2 nm.","Various materials may be chosen for producing the base of the cell according to the invention, in particular a material chosen from the group consisting of glass covered with a layer of gold, of cleaved mica, of silicon or of any other monocrystalline material.","Monocrystalline silicon is a very advantageous material insofar as it is effectively flat on an atomic scale, and it can readily be subjected to microetching and varied chemical modifications, which makes it possible to produce a support having a plurality of cells according to the invention.","Moreover, silicon has conducting or optical properties which may prove to be very advantageous as regards the detection of biological events taking place on the cell according to the invention.","As will be developed, the cell according to the invention may comprise several identical or different proteins linked to identical or different nucleic acids.","When different primers are used, a target nucleic acid complementary to two primers will be capable of binding the corresponding two floaters, which will accentuate its interaction with the support and will prime a change in spatial organization of the floaters within the cell.","Thus, when the nucleic acid IV-protein III and protein III-layer II bonds are reversible, it is possible, via agents which compete with the floater-support interaction (histidine, imidazole, EDTA), to readily differentiate the degrees of hybridization.","Moreover, the aspect of two-dimensional reorganization of the floater and primer nano-objects on the flat membrane surface, obtained through the fluidity of the layer II, falls within the area of the self-organization of biological molecules and of the physicochemical phenomena which ensue from these collective movements.","Thus, using a random distribution of the protein-nucleic acid blocks, it is clear that the action of an effector (for example a target molecule) will engender a radical restructuring in two dimensions.","When use is in particular made of a cell in which two (or more) different primers are present, the presence in the target sample of a nucleic acid which can hybridize to these various primers and which allows said blocks to become bridged should lead to a process of elongation (one-dimensional organization) at the surface of the cell.","There is therefore a change, and this is one of the major advantages of the invention, from a more or less disorganized system (random, or with an unstable equilibrium) to a highly ordered system, under the effect of the presence of the target nucleic acid in the sample tested.","Moreover, this organization is extremely sensitive to the geometric conformation of the nucleic acid and therefore to any factor (mismatching, protein binding, chemical agent) capable of interacting with the conformation or the structure of the nucleic acid.","Similarly, when the binding of any protein or of any ligand to a (single-stranded or double-stranded) nucleic acid linked to the protein anchored in the layer II is studied, or when the binding of an effector molecule on a protein (of the transcription factor type) linked to a nucleic acid, on a cell according to the invention, is studied, a modification of the structure (for example curvature of the nucleic acid) and of the equilibrium of the system are then observed (due to the introduction of these elements), which modifications can then be readily observed by various methods capable of detecting the organizational change in the cell, and in particular via an optical (absorbance, fluorescence), electrical, electron-based, surface plasmon resonance, energy transfer, radio-labeling, diffraction (optical, electrical, X-ray, neutron diffraction) or microscopy (direct optical or fluorescence microscopy, electron microscopy, near-field microscopy, atomic force microscopy) system.","This disorder-order phase change observed when an event occurs on an atomic scale can only be readily detected when the protein on the cell according to the invention has what has been called “an unequivocally detectable geometry” and when the nucleic acid is attached thereto unequivocally.","The principle of the invention is therefore the mobility of the protein anchored in the layer II, allowing a reorganization of the system, and the fact that the protein-nucleic acid unit is unequivocal and non-random, which makes it possible to detect said reorganization.","It is also envisioned that the presence of a single target nucleic acid molecule in the sample tested will make it possible to induce a reorganization which may then be detected.","An example of the implementation of such a method would be to pre-organize the cell with a nucleic acid of the same nature as that to be detected, but which, firstly, has a lower affinity for the primer than the nucleic acid to be tested (modified base, mismatch) and, secondly, has a geometry (size, curvature) which is different from the nucleic acid to be tested.","The exchange of a single nucleic acid (external competitor molecule) within a pre-organized cell will then lead to a local structural defect.","It is known by those skilled in the art that a single defect within a “crystalline” network destabilizes the entire network.","If the cell size is chosen in an appropriate manner, it is reasonable to hope that this single event will change the stability of the organization of the cell.","Such a change in stability can be tested by gradually applying an outside (chemical, temperature, electric potential, etc.)","factor which destabilizes the cell as far as disorganization.","This principle opens up the possibility of detecting single molecular events and corresponds to the theoretical maximum sensitivity of a detector.","This sensitivity is an enormous advantage compared to the other existing widely used systems, which require the presence of a large number of nucleic acid molecules in the sample in order to be detected.","Moreover, such a detector associated with a receptor protein chosen in an appropriate manner in order to see its interaction with the modulated DNA can be the basis of a molecular detector for an infinite variety of molecules of interest (in particular proteins, medicinal products, pollutants).","It is also important to note that the hybridization as such is not detected, but rather the changes induced in the cell according to the invention.","Thus, this implies that the cell according to the invention (or the support displaying a plurality of cells) can be used for detecting a target in a sample without it being necessary to label or to excessively handle said sample.","This is also a considerable advantage, since it is thus possible to detect the presence of mRNA without having to first convert it into cDNA.","It should be noted that the geometric conformation of ssDNAs and dsDNAs is so different that it results in very distinct physicochemical properties.","Thus, after hybridization, it is in particular possible to detect this effect by a change in the electronic conductivity of the cell according to the invention.","This is particularly easy when the protein according to the invention also has a redox center.","It is also possible to detect the hybridization by surface plasmon resonance, or by an optical method, which is facilitated by the presence of a protein exhibiting an anisotropic optical absorption.","In order to carry out the simultaneous detection of several targets, the invention therefore also relates to a support displaying a plurality of cells according to the invention.","The various cells may be separated from one another by microetching on the support, or by the presence, between two cells, of a hydrophobic layer consisting of elements with a chain length different from (preferably less than) the chain length for the elements of the layer I, for example less than or equal to 5 carbon atoms (“differential patterning” Cheng et al., 2000).","These elements having a low-carbon-number chain can be deposited onto the support using the masking techniques common in microelectronics.","The support according to the invention is advantageously completed by the setting up of a system for measuring the current, the impedance or the potential, making it possible to detect the events occurring on each one of the cells.","Another preferred alternative is that the support comprises a surface alternating conducting and reflecting zones and nonconducting and transparent zones.","Such a network comprising transparent zones clearly smaller in size than the wavelength of visible light has been described as having a function of light transmission according to wavelength in the form of a “comb” (Ebbesen et al., 1998).","The optical transmission properties of such a structure should be altered by the state of the neighboring cells, which provides a possible reading means.","It can also be envisioned that the support according to the invention comprises, between each cell, an integrated optical device (laser-photodiode couple for example) capable of measuring the absorbance of a cell in two cross directions and of deducing therefrom its state of organization (anisotropy) or of disorganization (isotropy).","It is also possible to preferably envision reading the organizational state of each cell via a laser optical scanning device sensitive to the change in light polarization.","Such a device capable of reading cells which are submicron in size is commonly used by the general public in minidisc readers and rewritable CD-Rom readers.","In such a case, the variation in polarization of the reflected light normally provided in magneto-optical discs by a reorganization of the crystalline network of a rare earth salt (europium, and other compounds of the same transition series of the Mendeleiev table) by a magnetic field and laser heating, would, in the system described in the invention, be provided by the variation in spatial organization, in the cell, of the protein or of its cofactor, which are objects which are optically active in terms of polarization, ellipticity (differential absorption of polarized light) and optical absorption.","The invention also relates to a method for identifying the presence of a test nucleic acid in a sample, comprising the steps of: a) bringing said sample into contact with a cell according to the invention, under conditions which allow the hybridization of said test nucleic acid to a nucleic acid attached to a protein of said cell, b) detecting the hybridization of said test nucleic acid to said nucleic acid attached to said protein.","The detection can be carried out by any means mentioned above.","If the test sample has been labeled, the detection can be carried out as a function of said labeling.","However, it may be preferred to detect the presence of the target nucleic acid by a change in conformation of the system.","Thus, the cell according to the invention allows the detection without there having been any need to label the test sample.","The invention also relates to a method for identifying the binding of a protein to a nucleic acid, and/or its activity on the conformation of said nucleic acid, comprising the steps of: a) bringing said protein into contact with a nucleic acid attached to a protein, in a cell according to the invention, b) detecting the binding of said protein to said nucleic acid, and/or its activity on the conformation of said nucleic acid.","Said nucleic acid may be single-stranded, double-stranded or triplex, when a nucleic acid has already been attached to one of the nucleic acids (primers) of the cell.","It is clear that the choice of certain bases for the primer can lead to the formation of a triplex nucleic acid, said formation then being considered, for the purpose of the invention, as a hybridization.","The detection can also be carried out in several ways, and in particular by a change in conformation of the system.","The invention also relates to a method for identifying the binding of a ligand to a protein attached to a nucleic acid attached to a protein in a cell according to the invention, comprising the steps of: a) bringing said ligand into contact with said protein, b) detecting the binding of said ligand to said protein.","Among the ligands and proteins envisioned, studies are in particular carried out in order to identify effector molecules which bind to nuclear receptors or transcription factors.","The binding may be direct or indirect, and may involve, for example, the binding of the ligand to a receptor which will then attach to a protein or a protein complex already associated with the DNA of the cell.","The binding of the ligand to the protein can be observed in particular by a change in conformation of the system.","In general, the invention relates to a method for identifying the binding of a compound to a nucleic acid, and/or its activity on the conformation of said nucleic acid, comprising the steps of: a) bringing said compound into contact with a nucleic acid attached to a protein, in a cell according to the invention, b) detecting the binding of said compound to said nucleic acid, and/or its activity on the conformation of said nucleic acid.","As above, said nucleic acid may be single-stranded, double-stranded or triplex, when a nucleic acid has already been attached to one of the nucleic acids of the cell.","The binding is in particular detected by a change in conformation of the system, as was explained above.","The detection of the geometry or of the disturbance of the geometry of the Floater-Primer-Target assemblies can advantageously be carried out through the contribution of atomic force microscopy technology in a liquid environment.","Thus, the potential applications of this type of characterization relate to studying the impact of the interaction of any molecules or macromolecules with nucleic acids (regulation, transcription factors, drugs, medicinal products, in particular for chemotherapy, etc.).","In a particular case, the device according to the invention offers a molecular and supramolecular proximity involving a lipid membrane, redox proteins and nucleic acids.","This assembly of electro-active and optically active biomolecules within microsystems derived from microelectronics may potentially prove to have a great impact in electron transduction (conductors, semi-conductors, biomolecular circuits) and nonlinear optical transduction (anisotropic) fields.","In fact, double-stranded DNA, unlike ssDNA, has properties of electron transfer according to a process of electronic conduction by overlap of the Pi orbitals.","In addition, the hybridization reaction between two ssDNA molecules is very specific.","The current systems developed in the context of electronic components on a nanometer scale come up against various problems, including: the interconnection of the nano-objects with one another, the connection of the nanostructures to macroscopic electrodes.","The use of processes of molecular recognition and of self-assembly of molecules within supramolecular structures, as envisioned within the cell according to the present invention, might make it possible to circumvent these difficulties.","In fact, the cell according to the invention exhibits an original biomolecular architecture which falls within the field of nano-bioengineering, i.e.","of the development of novel types of nanostructures having properties of: strong modularity, high degree of organization on a molecular scale, vectorial electron transfers, coupling with chemical messengers.","DNA has the mechanical molecular recognition properties for being integrated into microcircuits.","However, the intrinsic electronic properties of this macromolecule appear to be insufficient to be used directly (Braun et al., 1998).","One of the most promising areas of research in the use of electrical charge transport potential along the DNA duplex is represented by the use of electrocatalytic processes (Boon et al., 2000).","Such a signal transduction device can absolutely be integrated into the device according to the invention through the use of additional redox molecules.","However, in a preferred device according to the invention, the presence of floaters composed of redox proteins having electroactive groups and/or redox potentials which are different and which can be modulated makes it possible to provide an electron environment which is dense in terms of overlap of electron orbitals.","Depending on the periodicity and on the two-dimensional organization of these redox macromolecules (then considered to be relay centers), such a structure might give rise to electron doping of DNAs, to electron jumping between proteins or to energy transfer processes.","Thus, a cell according to the invention may have methods of use other than the simple methods of detection which have been mentioned above.","The invention also relates to a method for preparing a cell according to the invention, comprising the step of: introducing, on a support which is substantially flat on an atomic scale, a protein III having an unequivocal three-dimensional structure (stable and defined spatial organization), linked to a nucleic acid IV, such that the nucleic acid-protein assembly is laterally mobile relative to said support, in order for the hybridization of a target nucleic acid to said nucleic acid IV or the change in conformation of said nucleic acid IV to be detectable via the geometric reorganization of the nucleic acid-protein assembly.","The invention also relates to a method for preparing a cell according to the invention, comprising the steps of: attaching a hydrophobic monolayer I to a support which is substantially flat on an atomic scale, assembling, on said monolayer I, a mono-layer II comprising phospholipids, introducing a protein III having an unequivocal three-dimensional structure, said protein III being laterally mobile in said layer II, via a bridging molecule having a hydrophobic end capable of interacting with the monolayer II, and an end which has been chemically functionalized so as to form a stable bond with a protein III, said protein III being attached in an-unequivocal manner to a nucleic acid IV, preferably via a molecular arm.","DESCRIPTION OF THE FIGURES FIG.","1: Construction of the hybrid bilayer and characterization by surface plasmon resonance analysis A: The signal recorded corresponds to the base line for the metal support covered with a dense monolayer of OM in the environment of the working buffer (conventional phosphate buffer, 50 mM, pH 7.5).","B: The variation in signal corresponds to the injection into the fluid circuit of a DMPC-DOGS mixed vesicular solution produced in water.","The increase in the signal corresponds to fusion kinetics for the vesicles on the support so as to obtain a compact lipid monolayer.","Thus, after 20-40 min of injection, a return to the working buffer makes it possible to estimate the amount of lipids assembled at 1700±300 RU.","This value, according to the various modes of calibration proposed by BIAcore, corresponds to that of a lipid half-membrane.","C: A procedure for treating the membrane with a 20 mM sodium hydroxide solution makes it possible to remove the excess lipids adsorbed onto the structure.","D: Control of the homogeneity of the lipid membrane by injection of water-soluble protein (for example cytochrome c, BSA, etc.).","FIG.","2: Demonstration of a specific association of Hb5 (His)4 on the hybrid bilayer functionalized with chelating nickel cations Characterization by SPR A: Three successive injections (represented by arrows) of the cytochrome b5 polyhistidine form are carried out in the region of the functionalized lipid bilayer.","Conventionally, the protein solution, which has a concentration of 1 μM in 50 mM phosphate buffer, pH 7, is injected at a flow rate of 10 μl/min for 180 seconds.","The various injections produce a gradual saturation of the surface.","The maximum covering is of the order of 650±50 RU, i.e.","a degree of surface covering of 7 pmol.cm−2.B: In order to show the great specificity of the interaction, injections of a 1 mg/ml solution of histidine in 50 mM phosphate buffer are carried out (for example: four injections marked with arrows).","The injection of an agent which competes with the chelation between the polyhistidine segment of Hb5 (His)4 and the membrane.","Each injection (flow rate 10 μl/min for 120 seconds) is accompanied by a decrease in signal, i.e.","in the amount of proteins interacting with the membrane.","Thus, complete regeneration of the structure is obtained after a contact time of a few minutes.","FIG.","3: Diagram of the membrane biosensor, construction/regeneration of the supramolecular assembly Construction: An inorganic surface (for example: gold) is functionalized with a dense monolayer of octadecyl mercaptan.","This surface is suitable for the fusion of lipid vesicles (liposomes) until a dense monolayer of phospholipids is obtained above the support.","The presence of 10% (mol/mol) of modified phospholipids allows the anchoring of cytochromes b5 so as to obtain a dense monolayer of proteins.","Regeneration: Injections of agents which compete with the affinity via metal ions, such as a solution of histidine (1 mg/ml), make it possible to detach all the Hb5(His)4 interacting specifically with the support.","Similarly, the lipid half-membrane reconstituted on the functionalized inorganic support can be regenerated using a solution of detergent such as octyl glucoside, or OG (40 mM in H2O).","Thus, all of the constructing and destructuring phases can be carried out in a controlled manner using an aqueous solution which preserves the integrity of the biological molecules.","FIG.","4: Characterization of the b5-cis-Pt-ssDNA complex, SDS-polyacrylamide (15%) gel of Hb5(His)4 platinated and coupled with an oligonucleotide of sequence CTATCATTTGCTTACTATTC (SEQ ID No.","13) FIG.","4.A: Autoradiogram of a gel into which Hb5(His)4-cis-Pt has been injected; the 32P-radiolabeled oligonucleotide (well 2) and the b5-cis-Pt-ssDNA complex (well 3) make it possible to demonstrate the appearance of an additional radioactivity signal in well 3 compared to well 2.FIG.","4.B: Use of a polyacrylamide gel (15%) under denaturing conditions (SDS).","Cross-characterization of the b5-cis-Pt-ssDNA complex: said complex injected into lane 3 exhibits a band which makes it possible to confirm the existence of a population whose molecular weight is close to 20000 Daltons, i.e.","the theoretical molecular weight.","Well (1): b5 platinated with cis-Pt[(NH3)2(H2O)2]2+.","Well (2): radiolabeled 20-mer DNA (DNA*).","Well (3): platinated b5 linked to the 20-mer DNA* FIG.","5: Spectral characterization of the complex Hb5(His)4-CTATCATTTGCTTACTATTC via cis-Pt[(NH3)2(H2O)2]2+bridging Two procedures for reconstituting the Hb5 (his)4-cis-Pt-ssDNA complex are characterized by spectrophotometry in a 240-600 nm window.","The cytochrome b5 has an absorption maximum at 412 nm whereas the ssDNA has a maximum at 260 nm.","The spectral profile of two samples and also the relative quantification of each species by virtue of the molar extinction coefficients make it possible to demonstrate: red spectrum: an Hb5(His)4/ssDNA molar fraction= 3/1 blue spectrum: an Hb5(His)4/ssDNA molar fraction= 1/1 The procedure for the characterization represented by the blue absorption spectrum makes it possible to obtain equimolar coupling.","FIG.","6: Determination of the melting temperatures of the 20-mer DNA and of the 20-mer DNA bridged with platinated Hb5(His)4 The melting temperatures were determined by spectral measurements of hyperchromicity after melting of the complementary strand at high temperature (80° C.).","A decrease by steps of 1° C. makes it possible to follow the evolution of the dsDNA spectrally.","The results of absorption as a function of temperature for the free ssDNA makes it possible to obtain a curve whose inflexion point corresponds to the melting temperature.","The Tm for the ssDNA is evaluated at 45° C. The same characterization on the ssDNA coupled to Hb5(His)4 gives a modified profile, the Tm of which was estimated at 23° C. FIG.","7: Separation of the protein species and of the cytochrome b5-ssDNA complex via a DPDPB bridge, by chromatography.","Characterization by spectrophotometry at 240-450 nm The various populations derived from the incubation (4 H at 30° C.) of ssDNA-DPDPB with Hb5(His)4mut24 are loaded onto a DEAE column.","The various species are then eluted via a 25 mM phosphate solution and an increasing NaCl gradient.","The cytochrome b5 has an absorption maximum at 412 nm whereas the ssDNA has a maximum at 260 nm.","The NaCl gradient makes it possible to gradually elute all the populations: [NaCl]<0.15 M: population 1 of Hb5(His)4mut24 0.15<[NaCl]<0.25 M: population 2 of Hb5(His)4mut24 [NaCl]=0.5 M: population 3 of Hb5(His)4mut24 FIG.","8: Elution profile and supramolecular composition of the cytochrome b5-ssDNA complexes The molar quantification of the species isolated after separation on DEAE is represented in the form of a histogram: Populations 1 and 2 of Hb5(His)4mut24, characterized in FIG.","7, show no ssDNA component measured at 260 nm.","These two populations correspond to the monomeric form (which is not reacted during the incubation) and to the dimeric form (resulting from a cystine bridge between two proteins).","Population 3 shows an equimolar Hb5(his)4mut24-ssDNA complex.","FIG.","9: Detection by surface plasmon resonance of the kinetics of association of an ssDNA complementary to the primer ssDNA interacting with the biosensor Starting from a “b5-W1” signal in the region of 250 RU, various injections of ssDNA were carried out, at a b5-ssDNA density equal to 1 pmol.cm−2: Thus, the ssDNA strand complementary to W1 was injected in the region of the biosensor (1 μl/min for 10 min) for two concentrations of NaCl.","At a 150 mM concentration of NaCl, a hybridization signal is observed within a complexation range corresponding to 100% of the “b5-ssDNA” assemblies interacting with the membrane.","The same oligonucleotide injected at a low concentration of NaCl (25 mM) shows very restricted hybridization with respect to the biosensor.","FIG.","10: Diagram of the assembly as a ˜DNA biosensor The arrow represents the flow of biomolecules of the test sample.","FIG.","11: Diagram of the assembly as nanostructures with dynamic and conformational properties A set of various floating blocks can be displayed at the surface of the biosensor, and the injection of an oligonucleotide which potentially hybridizes on a part of the floating blocks should allow inter-block bridging.","The lateral mobility characteristic of the elements should promote this process until phenomena of supramolecular reorganization at the surface of the cell are obtained.","The following examples relate to a particular case of the invention, but should not be considered as limiting the invention.","EXAMPLES Example 1 Preparation of the Hybrid Bilayer 1.1) Functionalization of the Support The solid support used consists of a glass slide covered with a film of gold marketed by the company BIAcore® under the name chip J1.Such supports were also produced by the inventors.","They consist of the use of polished glass cover slips (thickness 0.4 mm, 22*10 mm, Prolabo) which are successively subjected to spraying of 2.6 nm chromium and then spraying of 48 nm of gold under a vacuum of 2×106 mmHg.","The metal interface is then covered by self-assembly of molecules of octadecyl mercaptan, or OM, from Aldrich.","The OM solution is used at 1 mM in a 4/1 (v/v) ethanol/water mixture.","After incubation for two hours, the slides are washed with chloroform, with methanol and then with water and are dried under a stream of nitrogen.","The result is the reconstitution on the gold of a highly organized dense monolayer which confers on the support very strong hydrophobic properties.","This first assembly has been characterized many times in the literature.","Contact angle measurements make it possible to routinely verify the compactness of the monolayer thus reconstituted.","1.2) Production of the Lipid Xesicles 1.2.1) Sonication/Extrusion Method The method used to prepare the liposomes is based on a sonication/extrusion double procedure.","The dimyristoyl and/or dipalmitoyl phosphatidyl-choline phospholipids from Sigma, which are the main constituents of the vesicles, are solubilized in an organic solution of chloroform.","A lipid film is then deposited in glass scintillation tubes by evaporation of the chloroform under a stream of nitrogen.","This film is resuspended in an aqueous solution (of ultrapure or buffered water) at a final concentration of 1 mM.","This mixture is then sonicated: in an ultrasound bath (Transsonic 310 from Fisher-Scientific) at maximum power for 20 min, and then with an ultrasonic disintegrator (Vibra-Cell from Bioblock) using 2 min pulses, with 50% active cycle for an outlet power of 4.The procedure is stopped when the solution has become translucent, i.e.","conventionally after 6 to 10 min of cumulative sonication time.","The final production step consists of extrusion through a polycarbonate membrane (Avestin, 19 mm in diameter and 50 nm or 100 nm in pore diameter) in a Liposofast® basic device, Avestin, Inc. 1.2.2) Vortex/Extrusion Method A second procedure is used when molecules are used which are sensitive to ultrasonic disturbances, such as the hydrocarbon chains of some phospholipids having polyunsaturations (for example: 1,2-dioleoyl-sn-glycero-3-[(N-(5-amino-1-carboxypentyl)iminodiacetic acid)succinyl] or DOGS, from Aventi Polar Lipids, and dioleoyl phosphatidylcholine, or DOPC, from Sigma).","In this case, a procedure identical to 1.2.1, consisting of solubilization in CHCl3 and then construction of a lipid film on glass, is applied.","The procedure then differs in that the phospholipids are resuspended in an aqueous solution at 1 mM via a step consisting of vortexing for 5 min at maximum rate.","This step is followed by a phase of extrusion through a polycarbonate membrane with a 50 nm pore diameter, in a Liposofast® device from AVESTIN.","1.3) Reconstitution of the Lipid Monolayer The hydrophobic support serving as matrix to generate the lipid monolayer is subjected to a treatment procedure as follows: in order to improve the wettability of the structure, an ethanol/water ( 1/1 vol) solution is injected onto the support and is followed by steps of rinsing with water and with buffered solutions.","In order to clean the interface, a solution of detergent, octyl glucopyranoside from Sigma, at a concentration of 40 mM, is injected.","Immediately following this, the lipid vesicles prepared are injected in contact with the functionalized metal surface.","The vesicles fuse spontaneously on contact with the hydrophobic monolayer of OM, according to a liposome fusion process established by Kalb et al.","(1992).","After 30 to 60 min, this process leads to the formation of a lipid monolayer completely covering the OM monolayer.","The molecular assembly thus obtained has a substrate which is metallic in nature, covalently covered with a first alkylated sheet on which is assembled a fluid and dynamic sheet of amphiphilic molecules.","The characterization experiments use the BIAcore® technology via BIAcore 1000 and BIAcore X devices.","The liposome fusion results are obtained in the form of a sensorgram (FIG.","1).","On injection of the vesicles, a phase of association with the support is observed, producing a plateau after a contact time of 20-30 [lacuna].","On stopping the injection, and therefore on returning to the working buffer, a first amount of lipid material is evaluated.","A procedure consisting of cleaning the surface with 20 mM sodium hydroxide makes it possible to remove all the vesicles incompletely fused at the surface of the biosensor.","Conventionally, the signal obtained is between 1400 and 2000 RU, which is in accordance with the results available in the literature and the methods of calibration provided by the manufacturer.","An injection of soluble proteins in the region of this supported membrane makes it possible to validate the absence of a defect in the upper lipid monolayer.","Introduction into the liposomes of synthetic DOGS phospholipids in a proportion of 10% mol/mol confers on this monolayer protein-HisTag sensor complexation properties.","Example 2 Construction, Expression and Purification of the Fusion Proteins 2.1) The Water-Soluble Proteins with a Histidine Tag The initial protein used in the context of this example is a membrane-bound heme protein: human or yeast cytochrome b5.It is a bitopic protein having a large globular water-soluble domain containing the heme and a small hydrophobic domain allowing anchoring to microsomal membranes.","All the nucleotide and amino acid sequences are included in the sequence listing.","The amino acid sequence of the human and yeast forms are referred to as follows: Amino Acid Sequence of Membrane-Bound Human Cytochrome b5 (SEQ ID No.","1) 1 MLAEQSDEAV KYYTLEEIQK HNHSKSTWLI LHHKVYDLTK FLEEHPGGEE VLREQAGGDA 61 TENFEDVGHS TDAREMSKTF IIGELHPDDR PKLNKPPETL ITTIDSSSSW WTNWVIPAIS 121 AVAVALMYRL YMAED Amino Acid Sequence of Membrane-Bound Yeast Cytochrome b5 (SEQ ID No.","2) 1 MPKVYSYQEV AEHNGPQNFW IIIDDKVYDV SQFKDEHPGG DEIIMDLGGQ DATESFVDIG 61 HSDEALRLLK GLYIGDVDKT SERVSVEKVS TSENQSKGSG TLVVILAILM LGVAYYLLNE The first experiment to modify the sequences of cytochrome b5 consisted in the removal, on the carboxy-terminal side, of the amino acids involved in the membrane insertion, so as to replace them with four histidines.","This chimeric or fusion protein is obtained by cloning the nucleotide sequence by polymerase chain reaction (PCR).","The cloning kit used is TOPO TA cloning® from Invitrogen.","The coding DNA sequence for the chimeric protein is introduced into a pCR®2.1-TOPO® vector.","The assembly was amplified in Vial of One Shot® cells.","The coding DNA sequence was then transferred into the PUHE25-2 expression vector (SphI cloning for the ATG and Bam HI for the stop).","The base modification for Hb5 is therefore as follows: Nucleotide Sequence of Membrane-Bound Hb5 (SEQ ID No.","3) 1 ATGGCAGAGC AGTCGGACGA GGCCGTGAAG TACTACACCC TAGAGGAGAT TCAGAAGCAC 61 AACCACAGCA AGAGCACCTG GCTGATCCTG CACCACAAGG TGTACGATTT GACCAAATTT 121 CTGGAAGAGC ATCCTGGTGG GGAAGAAGTT TTAAGGGAAC AAGCTGGAGG TGACGCTACT 181 GAGAACTTTG AGGATGTCGG GCACTCTACA GATGCCAGGG AAATGTCCAA AACATTCATC 241 ATTGGGGAGC TCCATCCAGA TGACAGACCA AAGTTAAACA AGCCTCCGGA AACTCTTATC 301 ACTACTATTG ATTCTAGTTC CAGTTGGTGG ACCAACTGGG TGATCCCTGC CATCTCTGCA 361 GTGGCCGTCG CCTTGATGTA TCGCCTATAC ATGGCAGAGG ACTGA Nucleotide Sequence of Soluble Hb5-Histidine Tag [Hb5-(His)4] (SEQ ID No.","4) 1 ATGGCAGAGC AGTCGGACGA GGCCGTGAAG TACTACACCC TAGAGGAGAT TCAGAAGCAC 61 AACCACAGCA AGAGCACCTG GCTGATCCTG CACCACAAGG TGTACGATTT GACCAAATTT 121 CTGGAAGAGC ATCCTGGTGG GGAAGAAGTT TTAAGGGAAC AAGCTGGAGG TGACGCTACT 181 GAGAACTTTG AGGATGTCGG GCACTCTACA GATGCCAGGG AAATGTCCAG AACATTCATC 241 ATTGGGGAGC TCCATCCAGA TGACAGACCA AAGTTAAACA AGCCTCCGGA AACTCTTATC 301 ACTACTATTG ATTCTAGTTC CAGTAACGGA CATCACCACC ATTAA Amino Acid Sequence of [Hb5-(His)4] (SEQ ID No.","5) 1 MLAEQSDEAV KYYTLEEIQK HNHSKSTWLI LHHKVYDLTK FLEEHPGGEE VLREQAGGDA 61 TENFEDVGHS TDAREMSRTF IIGELHPDDR PKLNKPPETL ITTIDSSSSN GHHHH 2.2) The Water-Soluble Protein-Histidine Tag with a Ser24→Cys mutation The chimeric protein obtained previously is then mutated in order to reveal a thiol group at the periphery of the protein, via a cysteine residue.","This modification is produced by virtue of a site-directed mutagenesis on serine No.","24 of Hb5(His)4.The mutagenesis kit used is the QuikChange® site-directed Mutagenesis Kit from Stratagene.","The action of the pfu DNA polymerase makes it possible to reconstitute the expression vector and the sequence to be mutated.","Supercompetent epicurian® XL1-blue bacteria are transformed with this plasmid.","Several different locations for the mutation were tried, and position 24 is in particular exemplified, i.e.","a modification of the serine residue to cysteine.","The mutation was verified by sequencing in the laboratory (ABI sequencer): Nucleotide Sequence of Hb5-(His)4 mut24 (SEQ ID No.","6) 1 ATGGCAGAGC AGTCGGACGA GGCCGTGAAG TACTACACCC TAGAGGAGAT TCAGAAGCAC 61 AACCACTGCA AGAGCACCTG GCTGATCCTG CACCACAAGG TGTACGATTT GACCAAATTT 121 CTGGAAGAGC ATCCTGGTGG GGAAGAAGTT TTAAGGGAAC AAGCTGGAGG TGACGCTACT 181 GAGAACTTTG AGGATGTCGG GCACTCTACA GATGCCAGGG AAATGTCCAG AACATTCATC 241 ATTGGGGAGC TCCATCCAGA TGACAGACCA AAGTTAAACA AGCCTCCGGA AACTCTTATC 301 ACTACTATTG ATTCTAGTTC CAGTAACGGA CATCACCACC ATTAA Amino Acid Sequence of Hb5-(His)4 mut24 (SEQ ID No.","7) 1 MLAEQSDEAV KYYTLEEIQK HNHCKSTWLI LHHKVYDLTK FLEEHPGGEE VLREQAGGDA 61 TENFEDVGHS TDAREMSRTF IIGELHPDDR PKLNKPPETL ITTIDSSSSN GHHHH 2.3) Protein Expression in Escherichia coli The bacterial expression strain is XL1-blue.","A preculture (6 h at 37° C. in 50 ml of medium) of a clone in a Luria-Bertami (LB) medium in the presence of a selection marker, ampicillin, made it possible to seed, in a proportion of 1/100, one liter of Terrific Broth medium supplemented with 100 mg/l of ampicillin.","The culture is placed at ambient temperature with shaking for 48 h and then overexpression of protein is induced by adding isopropyl-β-D-thiogalactopyranoside (IPTG) in a final proportion of 0.5 mM.","2.4) Protein Purification The bacteria are centrifuged after induction for 24-48 h and are frozen.","A freezing/thawing cycle is followed by a lysis/fractionation step (lysozyme, cholic acid and ultrasonic disintegrator).","The metal ion affinity chromatography (MIAC) purification procedure is defined as follows: An agarose gel functionalized with iminodiacetic acid (Sigma) is pre-loaded (flow of 1 ml/min) with nickel by loading a 50 mM solution of nickel chloride solubilized in an acetate buffer, pH 5.4.The column is washed with this same buffer at the same flow and the acetate is then replaced with a 50 mM sodium phosphate buffer, pH 7.The cell fractionation is directly injected onto the agarose gel column at 5 ml/min.","Specifically, the chimeric proteins are retained on the column according to the MIAC technique.","The column is subjected to a washing phase with phosphate buffer of increasing concentration of 50 to 250 mM, pH 7, until the solution passing through no longer exhibits an absorption spectrum in the ultraviolet range (240-300 nm).","The protein sample is eluted during pulses of histines in solution at a concentration of 1 mg/ml in the 50 mM phosphate buffer, pH 7.Similar experiments were carried out on the yeast form of cytochrome b5.Thus, it is possible to obtain several mutant human and yeast chimeric proteins with a high degree of purity and in a proportion of several milligrams of protein.","Example 3 Production of the Floating Protein Anchoring The hybrid bilayer structure presented in paragraph 1.3 was functionalized with the aim of conferring on the model metal ion affinity properties.","To do this, a synthetic phospholipid having an imino-diacetate residue which complexes a nickel divalent cation was chosen.","It is DOGS, or 1,2-dioleoyl-sn-glycero-3-[(N-(5-amino-1-carboxypentyl)iminodiacetic acid)succinyl] from Avanti polar Lipid, Inc.","It has been demonstrated that such a molecule allows the interaction of proteins having a polyhistidine domain in their primary sequence.","With the aim of introducing these synthetic phospholipids into the membrane model, we produced lipid vesicles composed of a mixture of lipids containing 10% (mol/mol) of modified lipid.","These mixed liposomes have the same behavior with respect to the hydrophobic support as those composed of a single species of phospholipids.","The fusion results by SPR give rise to values which are comparable in terms of kinetics of fusion and of covering (results not shown).","A method for addressing soluble proteins to membrane supports in a stable and specific manner is to use the coupling between a chelating divalent metal cation present at the surface of the membrane and histidine residues.","It has been demonstrated that cytochrome b5 in which the membrane domain has been deleted in favor of a histidine Tag segment conserves an affinity for biological membranes displaying chelating divalent cations.","Various forms of cytochrome b5 with a HisTag domain were produced (paragraph 2.1).","The immobilization of the cytochromes b5 on the hybrid bilayer via an MIAC approach was characterized by SPR (FIG.","2).","To evaluate the binding capacity of the membrane, multiple protein injections were performed within its surroundings.","The protein concentrations are conventionally between 0.5 and 2 μM and produce biosensor saturation kinetics.","For the Hb5(His)4 form, a maximum load signal of 650 RU, which corresponds to approximately 75% of optimum covering on such a membrane model (FIG.","1a), was obtained.","To demonstrate the specific interaction of these proteins with the biosensor, multiple injections of an agent which competes with histidine-Ni2+ coordinations were performed.","A solution of histidine at 1 mg/ml injected at the surface of the biosensor produces a large decrease in the signal, i.e.","in the amount of cytochrome b5 attached to the membrane (FIG.","2b).","In this way, it is possible to compete with the specific interaction and to return to the signal base line.","This procedure, using an aqueous solution which is nondenaturing for the biomolecules, thus enables complete regeneration of the surface of the lipid monolayer.","The results given in FIG.","2 confirm the strong potential of the biosensor in terms of protein capture with a high degree of compactness.","They make it possible to underline the very high level of control of the authors over the various phases of construction of this supramolecular assembly since, by modulating the protein or competitor injection pulses, it is possible to precisely attach the amount of protein at the surface.","This great modularity can be summarized in FIG.","3.Example 4 Protein-ssDNA Grafting Via a Cis-Pt Complex Platinum complexes, in particular cis-diamine-dichloroplatinums (cis-[PtCl2(NH3)2]) are agents which bind covalently to DNA.","In addition, these molecules, by substitution of their labile chloro ligand, can couple, firstly, a base of a DNA and, secondly, a nucleophilic residue of an amino acid of a protein.","4.1) Materials Cis- [PtCl2(NH3)2] and trans- [ PtCl2 (NH3)2] are provided by Johnson Matthey (London, United Kingdom).","Cis-[Pt(NH3)2 (H2O)2] 2+ and trans- [Pt(NH3)2 (H2O)2] 2+are prepared by dissolving a suspension of cis-[Pt(NO3)2 (NH3)2] and trans- [Pt(NO3)2 (NH3)2] in water, respectively formed by reacting cis-[PtCl2(NH3)2] and trans-[PtCl2(NH3)2] with silver nitrate.","The 20-mer DNA, its complementary strand and its biotinylated complementary strand come from Eurogentec.","The sequence of the 20-mer DNA, 5′CTATCATTTGCTTACTATTC 3′(SEQ ID No.","13), is chosen such that it possesses only one guanine since the N7 nitrogen is the main site of platination of DNA (Lepre et al., 1990).","4.2) Formation of the DNA-b5 Bridging Via Platinum Bridging The platination reactions are carried out firstly on the DNA or on the cytochrome b5 (Hb5(His)4 and Hb5(His)4mut24), but the DNA-b5 bridging yield is greater when the cytochrome b5 is platinated first.","Platination of the ssDNA (500 μM) with one equivalent of platinum complex in 70 mM NaClO4 leads to the formation of the cis- or trans- [Pt(NH3)2(H2O) ]2+ monoadduct linked to the guanine.","The platination site is determined after treatment of the modified oligonucleotide with piperidine-DMS under “Maxam-Gilbert” sequencing conditions followed by treatment with NaCN.","A lack of reactivity of the platinated N7 guanine with the dimethyl sulfate (DMS) makes it possible to conclude that the latter is protected with respect to the DMS and is therefore effectively platinated (Comess et al., 1990).","The cytochrome b5 (100-200 μM) is incubated in the presence of 5 equivalents of platinum complexes in unbuffered water (pH 5) for 5 hours at 25° C. The cytochrome b5 is then incubated (90 μM) with 1.5 equivalents of DNA in unbuffered water (pH 5) for 50 hours at 25° C. The DNA-b5 complexes are separated from the non-complexed DNA molecules using an agarose gel functionalized with Ni2+/iminodiacetic acid groups (Sigma).","The protein is eluted by injection of imidazole (1 M).","The excess DNA is not retained on the column.","The DNA-b5 complexes coupled via a platinum bridge are also purified from the nonreactive forms of DNA using streptavidin-coated magnetic spheres from Promega, according to a modification of the procedure described by Promega: 0.5 nmol of total DNA (complexed or non-complexed) of the complexation mixture is heated at 45° C. for 10 min in water and is then brought into contact with its biotinylated complementary strand in 0.075 M NaCl, 0.0075 M citrate, pH 7.2, at 2.5° C. The mixture (500 μl) is added to 0.6 mg of magnetic spheres complexed with streptavidin, pre-washed and equilibrated in the same buffer at 2.5° C. The ssDNA-b5 platinum complexes are not retained on the spheres and the non-complexed ssDNA is eluted between 30-40° C. in 0.015 M NaCl and 0.0015 M citrate, pH 7.2.These purification steps are followed, firstly, by detection of the radioactivity of the ssDNA labeled with 32P in the 5′ position (radiolabeled beforehand via the action of polynucleotide kinase using [γ32P]ATP) and, secondly, by a spectral study over a window of absorbance comprising 413 nm for the b5 (e=100 000 M−1.cm−1) and 260 nm for the ssDNA (e=180500 M−1.cm−1) .","The latter is corrected for the Hb5 absorbance component at this wavelength (i.e.","⅕ of its absorbance at 413 nm).","However, it is specified that these last two procedures do not make it possible to separate the “ssDNA-b5” platinum complex from the non-complexed cytochrome b5.The cis- [Pt(NH3)2(H2O)2]2+ and trans-[Pt(NH3)2(H2O)2]2+ platinum complexes are capable of coupling the cytochrome b5 with the 20-mer ssDNA.","The platination site on the DNA oligonucleotide occurs at N7 of a unique guanine.","The “DNA-b5” macromolecular complexes are characterized by gel electrophoresis analyses under denaturing conditions, spectro-photometric quantification analyses and determination of melting temperature with a complementary free ssDNA.","An electrophoretic study on SDS-15% PAGE of the mixture composed of the 5′-radiolabeled ssDNA, of Hb5[His]4 and of cis- or trans- [Pt(NH3)2(H2O)2] 2+was carried out.","After migration of the samples, the radioactivity of the ssDNA is associated with the Hb5[His]4 migration band (revealed by coomassie blue staining), demonstrating that a covalent bond has been formed between the 20-mer ssDNA and the cytochrome b5 (FIG.","4).","The “DNA-pt-b5” complex was characterized by spectral analysis with the aim of determining the stoichiometry of the reaction between the cytochrome b5 and the 20-mer ssDNA.","The experimental conditions allow the formation of an equimolecular complex as indicated by the respective absorbance of the cytochrome b5 at 413 nm and of the 20-mer ssDNA at 260 nm (FIG.","5).","4.3) Determination of the Melting Temperature The melting temperatures of the 20-mer ssDNA and of the 20-mer ssDNA coupled to the b5 via a cis-[Pt(NH3)2 (H2O) 2] 2+bridge were determined in 0.1 M NaClO4, pH 4, with their complementary ssDNAs.","The melting temperatures were measured on a Uvikon 941 spectrophotometer (Kontron).","0.2 nmol of 20-mer ssDNA (or “ssDNA-b5-Pt” complex) were incubated with an ssDNA having a complementary sequence, at a final concentration of 0.5 μM, in 0.025 M NaClO4, pH 4.4.The duplex sample is heated to 80° C. (20-mer DNA) or 65° C. (“DNA-Pt-b5”), the temperature is subjected to a decrease of 1° C./min down to 2.5° C., and the absorbance is measured at 260 nm.","The samples are heated from 2.5° C. to 60° C. at 1° C./min and their absorbance is followed at 260 nm, with the aim of verifying that the curves obtained during the cooling and the heating can be superimposed (reversible profiles).","The Tm values were determined using Kontron software.","The melting temperature of the duplex containing the 20-mer ssDNA decreases by 22° C. (from 45° C. to 23° C.) when it is coupled with the cytochrome b5 (FIG.","6).","No difference was observed between the complex comprising Hb5(His)4 and the cysteine mutant; the cysteine at position 24 does not appear, for the cis-Pt complex, to be more reactive than the other nucleophilic sites present at the periphery of the cytochrome b5.Be that as it may, the platination reactions do not occur on the 4-histidine tag in the carboxy-terminal position, in which case the complex thus formed could not be purified on an agarose-imino-diacetate column or would not be capable of interacting strongly with the cell.","Experiments for characterizing the various complexes by mass spectroscopy (electrospray, Maldi-TOF) are in the process of being carried out.","Example 5 Protein-ssDNA Grafting Via a Spacer Arm 5.1) Functionalization of ssDNA with DPDPB A second alternative was explored, concerning the grafting of a nucleic acid with the cytochrome b5.With a view to optimum display of the DNA coupled at the surface of the membrane biosensor, the inventors chose to use not direct coupling, but preferentially complexation via a spacer arm.","The molecule providing the intermolecular bridging is 1,4-di-[3′-(2′-pyridyl-dithio)propionamido]butane, or DPDPB.","This molecule allows bonding of the covalent type between two molecules having thiol functions and provides an intermolecular space of 1.6 nm.","In order to produce such a supramolecular assembly, a thiol function was created both on an oligonucleotide and on the cytochrome b5.The modified oligonucleotides are of HPLC quality and are produced by Eurogentec.","The various modified oligonucleotides are as follows: Oligonucleotide No.","W1 (SEQ ID No.","8) 5′GCT-AGC-TGC-ATA-GAT-CTC-TAC-C-SH 3′ The thiol modification is performed at the 3′ end of the 22-mer.","Oligonucleotide No.","W2 (SEQ ID No.","9) 5′ GCT-AGC-TGC-ATA-GAT-CTC-TAC-C 3′ The thiol modification is performed intra-chain on a thymine placed at position 11 of the 22-mer.","The various nonmodified oligonucleotides involved in the exemplification are as follows: Oligonucleotide No.","W3 (SEQ ID No.","10) 5′ GGT-AGA-GAT-CTA-TGC-AGC-TAG-G-3′ W3 is the 22-mer complementary to W1.Oligonucleotide No.","W4 (SEQ ID No.","11) 5′-GCA-GCT-AGC-GGT-AGA-GAT-CT-3′ W4 has a sequence complementary to the first 9 bases of Wl (5′ side) and to the first 11 bases of W1 (3′ side).","In this manner, W4 has the particularity of being able to bridge two W1 oligonucleotides.","Oligonucleotide No.","W5 (SEQ ID No.","12) 5′-GCT-AGC-TGC-ATA-GAT-CTC-TAC-C-3′ W5 corresponds to the random base sequence of W1.The DPDPB-W1 coupling procedure is as follows: in order to prevent the dimerization of the W1s by disulfide bonding, the oligonucleotides are treated with a dithiothreitol (DTT) reducing agent, in a molar fraction proportion of ⅕.","The mixture is incubated at 30° C. for 15 min.","The stock solution of DPDPB is solubilized in dimethyl sulfoxide (DMSO) in a proportion of 10 mg/ml.","DPDPB, in a proportion of 1 mol of W1 per 500 to 5 000 mol of DPDPB, is added in the presence of the residual DTT, for an incubation time of 2 to 4 hours at 30° C. The W1 sample is then specifically attached to a DEAE ion exchange column, the reagents are eliminated, and the oligonucleotide is eluted by adding a 1.5 M NaCl solution.","The degree of modification can be calculated by assaying the cross-linking agent with DTT.","This reduction causes the second thiopyridine group of the bridging agent to leave, and it can be quantified spectrally at 341 nm.","The procedure described makes it possible to achieve 100% of complex.","5.2) Cytochrome b5-W1 Grafting Via the Spacer Arm In order to perform controlled and reversible grafting on the cytochrome b5, the inventors carried out a site-directed mutagenesis of Hb5(His)4 as described in paragraph 2.2, producing the protein Hb5(His)4mutant24.The serine→cysteine mutation makes it possible to specifically introduce a thiol group at the periphery of the protein.","In order to prevent dimerization of the proteins, the solution is treated with DTT (Xmol b5/DTT=⅕) for 30 min at 30° C. The sample is then loaded onto a gel exclusion chromatography column (G25) so as to remove the DTT and is immediately injected into the solution of modified oligonucleotides (Xmol b5/W1=½) for 4 h at 30° C. The excess oligonucleotide is removed by retention of the proteins on an iminodiacetate-Ni2+gel.","Elution of the proteins is obtained by pulses of a 1 mg/ml histidine solution, as indicated during the HisTag protein purification step.","Spectral analysis over a [240-500] nm window makes it possible to demonstrate the effective coupling of Hb5(His)4mut24 with W1 (FIG.","7).","With regard to the molar absorption coefficients of the two macromolecules, the degrees of complexation range from 35 to 65%.","In order to subsequently use only an equimolecular Hb5/W1 population, an additional purification step is carried out on DEAE.","Elution performed under an NaCl concentration gradient makes it possible to isolate the various species reconstituted during the coupling (monomeric and dimeric Hb5, Hb5-spacer-W1).","Thus, all the species are separated from the initial population, by an increasing NaCl gradient (FIG.","7): Spectrum a) Hb5(His)4mut24 monomer Spectrum b) Hb5(His)4mut24 dimer Spectrum c) equimolar b5-W1 complex.","According to this procedure, the b5-W1 population is isolated and can be brought into contact with the hybrid membrane.","Example 6 Preparation of the Supramolecular Assembly “b5-cis Pt-ssDNA” and “b5-DPDPB-ssDNA” The grafting of the various complexes onto the hybrid bilayer was characterized by surface plasmon resonance in a BIAcore device.","Radioactivity-based and fluorescence labeling-based experiments are in the process of being developed in the laboratory.","The experiments consisting of coupling the molecular assembly to the lipid model by virtue of the chelating nickel complex by surface plasmon resonance made it possible to show the formation of a stable complex.","The levels of compactness obtained with the assembly appear, on the other hand, to be less than those obtained with the protein not modified with the oligonucleotide.","The increase in negative charges introduced by W1 may be responsible for repulsions between supramolecular blocks and may thus lead to less covering.","The covering values were evaluated at between 200 and 250 RU.","It was verified that, like Hb5(His)4, the structures are interacting specifically with the support and that the action of agents which compete with the chelation, such as a 1 mg/ml histidine solution, allows complete regeneration of the structure (data not shown, but exhibiting the same behavior as those appearing in FIG.","2).","Example 7 Potential of the Biosensor and Detection of Hybridization In order to evaluate the potential of the membrane biosensor in terms of specificity and sensitivity, various tests were carried out.","Firstly, it was essential to verify the absence of nonspecific interactions of oligonucleotides with the biosensor.","To do this, various injections of W4 were performed.","W4 represents a random sequence from the bases constituting W1; studying the evolution of the signal made it possible to show that W4 does not interact at the surface of the biosensor, i.e.","the lipid structure and the b5-W1 supramolecular assembly, whatever the bridging means (cis-Pt or DPDPB).","Secondly, various injections of W3 were performed at various salt concentrations.","Thus, it was clearly demonstrated that the injections of W3, the sequence complementary to W1, in the presence of a high concentration of NaCl were accompanied by association or hybridization kinetics.","On the other hand, for concentrations of NaCl less than or equal to 20 mM, the same oligonucleotide interacts very little or not at all with the biosensor.","These results demonstrate the possibility of detecting hybridization states between two ssDNAs at the surface of the biosensor.","The hybridization results show a threshold value of approximately 30 to 35 RU.","Now, according to the various methods of calibration, the value expected for complete hybridization based on a b5-W1 block signal of 250 RU is 70 RU.","However, the Tm measurements made on W1 clearly show a phase transition temperature close to ambient temperature, i.e.","the temperature at which measurements are made in the BIAcore device.","At this temperature, only 50% of the hybridization is expected.","The value measured with our device is therefore close to optimum hybridization, which is proof of a very favorable accessibility of the target molecules displayed by the biosensor.","The size of the working cell within the BIAcore device is of the order of 1 mm2.However, the surface really probed by the damped wave represents only 0.224 mm2 Considering the calibration 1 000 RU←→0.224 ng/0.224 mm2, 250 RU of molecular assembly correspond to 0.1 ng, i.e.","5 fmol ≈3×109 molecules.","The membrane biosensor allows the molecular hybridization of approximately 109 molecules.","In addition, the biosensor presented requires only the capture of a complementary ssDNA, without it being necessary to use a labeled probe.","Bibliographical References Berney et al.","(2000) Sensors amd Actuators B 68, 100-108 Boncheva et al.","(1999) Langmuir 15, 4317-4320 Boon et al.","(2000) Natural Biotechnology 18, 1096-1100 Braun et al.","(1998) Nature 391, 775-778 Cheng et al (2000) Reviews in molecular biotechnology 74, 159-174 (p 18) Comess et al.","(1990) Biochemistry, 29, 2102-2110 Ebbesen et al.","(1998) Nature 391, 667-669 (p 18) Gulik-Krzywicki et al.","(1967) J Mol Biol; 27(2): 303-22 Heyse et al.","(1998) Biochemistry 37, 507-522 Kalb et al.","(1992) Biochim.","Biophys.","Acta 1103: 307-316 Lepre et al.","(1990 ) Nucleic Acids Molec.","Biol., 4, 9-38 Marchal et al.","(1998) Biophysical Journal 74, 1937-1948 Steel et al.","(2000) Biophysical Journal 79, 975-981 Wittung-Stafshede et al.","(2000) Colloids and Surfaces 174, 269-273"]],"string":"[\n [\n \"Supported double layer structure for displaying a nucleic acid associated with a protein\",\n \"The invention concerns novel biosensors, in particular a support for displaying nucleic acids and for detecting both the presence of nucleic acids in a sample and the linkage between proteins and nucleic acids, as well as the linkage between a ligand and a protein linked to a nucleic acid.\"\n ],\n [\n \"1.A cell for displaying a nucleic acid, comprising: a support which is substantially flat on an atomic scale, a protein III having an unequivocal three-dimensional structure, a nucleic acid IV linked to said protein III, the nucleic acid-protein assembly being laterally mobile relative to said support, such that the hybridization of a target nucleic acid to said nucleic acid IV or the change in conformation of said nucleic acid IV can be detected via the geometric reorganization of the nucleic acid-protein assembly.\",\n \"2.The cell as claimed in claim 1, characterized in that said protein III is chosen from the group consisting of proteins having a redox center and proteins having an anisotropic optical absorption property.\",\n \"3.The cell as claimed in one of claims 1 and 2, comprising: a support which is substantially flat on an atomic scale, to which is attached a hydrophobic monolayer I, on which is present a monolayer II comprising phospholipids, a bridging molecule having a hydrophobic end which can interact with the monolayer II, and an end which is chemically functionalized so as to form a stable bond with a protein III, a protein III having an unequivocal three-dimensional structure, said protein III being laterally mobile in said layer II, a nucleic acid IV unequivocally attached to said protein III, preferably via a molecular arm.\",\n \"4.The cell as claimed in one of claims 1 to 3, characterized in that the nucleic acid is attached to said protein via a covalent bond.\",\n \"5.The cell as claimed in one of claims 1 to 3, characterized in that the nucleic acid is attached to said protein via a reversible bond.\",\n \"6.The cell as claimed in one of claims 1 to 3, characterized in that the nucleic acid IV is attached to said protein III via a compound chosen from: a spacer arm connecting two identical or different, chemically reactive residues of the nucleic acid IV and of the protein III, chosen from the group consisting of thiols, amines, amides and arginines; a metal complex such as cis-platinum, trans-platinum, europium, or a complex of nickel, of copper or of ruthenium coordinated, firstly, with an amino residue or a group of amino residues of said protein III and, secondly, with one or more natural or modified bases of the nucleic acid IV, a covalent or noncovalent protein complex involving the nucleic acid IV.\",\n \"7.The cell as claimed in one of claims 1 to 6, characterized in that said protein III has undergone a modification which introduces a unique site for effecting said bond [lacuna] said nucleic acid IV in an unequivocal manner.\",\n \"8.The cell as claimed in claim 7, characterized in that said modification consists of a mutation of said protein so as to leave remaining or to introduce just one unique predetermined amino acid.\",\n \"9.The cell as claimed in claim 8, characterized in that said predetermined amino acid is a cysteine.\",\n \"10.The cell as claimed in one of claims 3 to 9, characterized in that said protein is attached to a phospholipid of the layer II.\",\n \"11.The cell as claimed in claim 10, characterized in that said bond is a covalent bond, for example via acylation, farnesylation, via a GPI (glycosyl-phosphatidylinositol) anchoring sequence or via the attachment of a phospholipid to a specific residue of the protein III by any artificial method.\",\n \"12.The cell as claimed in claim 10 or 11, characterized in that said bond is a reversible bond, for example via an arm capable of linking a metal chelate on one side and an amino acid or a group of amino acids on the other.\",\n \"13.The cell as claimed in one of claims 10 to 12, characterized in that said protein is linked to a modified phospholipid of the layer II, said phospholipid having a chelate-binding zone.\",\n \"14.The cell as claimed in one of claims 3 to 9, characterized in that said protein has a hydrophobic tail which allows it to be anchored in the layer II.\",\n \"15.The cell as claimed in one of claims 3 to 14, characterized in that the phospholipids of the layer II have hydrophobic tails chosen such that said layer is not in the crystalline state, but in the fluid state at the temperature for use.\",\n \"16.The cell as claimed in claim 15, characterized in that said hydrophobic tails are greater than or equal to 14 carbon atoms in chain length, said chain possibly having unsaturations, such that said layer II is in the fluid state at the temperature for use.\",\n \"17.The cell as claimed in one of claims 3 to 16, characterized in that said phospholipids of the layer II have polar heads, chosen from the group consisting of neutral polar heads carrying no charges, polar heads which are neutral overall but which carry opposite charges (zwitterionic polar heads), or polar heads carrying an overall negative charge.\",\n \"18.The cell as claimed in one of claims 3 to 17, characterized in that said layer I comprises hydrophobic elements with a chain length of approximately 2 to 2.5 nm, for example of between 12 and 20 carbon atoms, preferably between 14 and 18 carbon atoms.\",\n \"19.The cell as claimed in one of claims 3 to 18, characterized in that said layer I comprises phospholipids linked to said support via their polar heads.\",\n \"20.The cell as claimed in one of claims 3 to 18, characterized in that said layer I comprises hydrophobic elements linked to said support via a covalent bond, for example an alkyl-thiol bond or an alkyl-siloxane bond.\",\n \"21.The cell as claimed in one of claims 1 to 20, characterized in that the flatness of said support is such that the difference in height between two zones separated by less than 100 nm is less than or equal to 10 nm, preferably less than or equal to 3 nm, more preferably less than or equal to 2 nm.\",\n \"22.The cell as claimed in one of claims 1 to 21, characterized in that said support consists of a material chosen from the group consisting of glass covered with a layer of gold, cleaved mica, silicon or any other monocrystalline material.\",\n \"23.The cell as claimed in one of claims 1 to 22, characterized in that it comprises several identical or different proteins linked to identical or different nucleic acids.\",\n \"24.A support for detecting nucleic acids, displaying a plurality of cells as claimed in one of claims 1 to 23.25.The support as claimed in claim 24, characterized in that it comprises a system for measuring current, impedance or potential.\",\n \"26.The support as claimed in claim 24, characterized in that it comprises, between each cell, an integrated optical device capable of measuring the absorbance of a cell in two cross directions and of deducing therefrom its state of organization (anisotropy) or of disorganization (isotropy).\",\n \"27.The support as claimed in claim 24, characterized in that it comprises a surface alternating conducting and reflecting zones and nonconducting and transparent zones.\",\n \"28.A method for identifying the presence of a test nucleic acid in a sample, comprising the steps of: a) bringing said sample into contact with a cell as claimed in one of claims 1 to 23, under conditions which allow the hybridization of said test nucleic acid to a nucleic acid attached to a protein of said cell, b) detecting the hybridization of said test nucleic acid to said nucleic acid attached to said protein.\",\n \"29.A method for identifying the binding of a protein to a nucleic acid, and/or its activity on the conformation of said nucleic acid, comprising the steps of: a) bringing said protein into contact with a nucleic acid attached to a protein, in a cell as claimed in one of claims 1 to 23, b) detecting the binding of said protein to said nucleic acid, and/or its activity on the conformation of said nucleic acid.\",\n \"30.A method for identifying the binding of a ligand to a protein attached to a nucleic acid attached to a protein in a cell as claimed in one of claims 1 to 23, comprising the steps of: a) bringing said ligand into contact with said protein, b) detecting the binding of said ligand to said protein.\",\n \"31.A method for identifying the binding of a compound to a nucleic acid, and/or its activity on the conformation of said nucleic acid, comprising the steps of: a) bringing said compound into contact with a nucleic acid attached to a protein, in a cell as claimed in one of claims 1 to 23, b) detecting the binding of said compound to said nucleic acid, and/or its activity on the conformation of said nucleic acid.\",\n \"32.The method of identification as claimed in one of claims 28 to 31, characterized in that the detection is carried out via an optical (absorbance, fluorescence), electrical, electron-based, surface plasmon resonance, energy transfer, radiolabeling, diffraction (optical, electron, X-ray, neutron diffraction) or microscopy (direct optical or fluorescence microscopy, electron microscopy, near-field microscopy, atomic force microscopy) system.\"\n ],\n [\n \"The present invention relates to novel biostructures which can in particular be used as biosensors, in particular a support for displaying nucleic acids and for detecting both the presence of nucleic acids in a sample and the interaction between proteins and said nucleic acids, and also the binding between a ligand and a protein linked to a nucleic acid.\",\n \"The development of DNA biosensors, also referred to as DNA chips, has considerably increased over the last few years.\",\n \"The impact of such biosensors has mainly been directed toward sequencing, gene expression mapping, diagnosis (detection, damage, mutation, etc.)\",\n \"and analysis of DNA-ligand interactions (hybridization, etc.).\",\n \"Most of the biosensors employed use techniques for immobilizing single-stranded nucleic acid, or ssDNA, molecules, the analysis being carried out by virtue of optical transduction, electrochemical or piezoacoustic systems.\",\n \"The fact that biological membranes are highly important in physiological terms as a site for numerous chemical and metabolic reactions has led to a craze for developing lipid models on inorganic surfaces.\",\n \"The design of such biocompatible and biofunctional structures on solid substrates has made it possible to develop models involving interdisciplinary fields which have brought about an expansion of many lines of scientific research and also practical applications.\",\n \"The main applications have thus concerned: the production of matrices for immobilizing enzymes, cell receptors or hormones in nondenaturing environments, the design of interfaces for capturing cells in order to study the effects of pharmaceutical molecules on the outcome of the cell, the self-assembly of very resistant membranes for the development of biosensors.\",\n \"The analytical methods applied to the analysis of DNA-DNA biomolecular interactions, such as they are represented at the heart of DNA chip devices, are essentially based on the attachment of a single-stranded DNA (ssDNA) within a delimited area on a substrate.\",\n \"The main techniques for producing DNAs covalently attached to the substrate are the in situ synthesis of ssDNA on the support, the assembly of ssDNAs which have been chemically modified to allow grafting onto the support, or microdeposition onto an adsorbent matrix (Boncheva et al., 1999, Berney et al., 2000, Wittung-Stafshede P. et al., 2000).\",\n \"Now, in the development of biosensors, the arrangement of biomolecules at the interfaces plays a crucial role.\",\n \"In order to obtain highly sensitive surfaces, it is important to exhibit receptor molecules in such a way that the corresponding ligand can interact without steric hindrances.\",\n \"However, in these various approaches, the means used to produce these DNA matrices generate artifacts (errors in the in situ syntheses or in functionalization of oligonucleotides) which cause nonspecific adsorptions; thus, some of the current studies relate to means for getting round these various limitations (Steel et al., 2000).\",\n \"Another limitation of these systems is inherent to the method of displaying the ssDNAs.\",\n \"The highly compacted nature of the molecules on the surface leads to the appearance of steric constraints which affect the hybridization efficiency of the chip.\",\n \"While the immobilization of ssDNAs on a surface is to date the most commonly used alternative, it also constitutes that which restricts the geometry of the device, resulting in a complete loss of flexibility and of modularity of the assembly.\",\n \"One of the characteristics of the present invention is to provide a novel method of displaying nucleic acids at the surface of the substrate.\",\n \"The method of displaying the nucleic acids according to the invention is original in that it allows the nucleic acids to be mobile, and thus makes it possible to detect any change in the state of said nucleic acids by detecting a transition between two states exhibiting distinct organizational levels.\",\n \"Thus, the subject of the present invention is a cell for displaying a nucleic acid, comprising: a support which is substantially flat on an atomic scale, a protein III having an unequivocal three-dimensional structure (stable and defined spatial organization), a nucleic acid IV linked to said protein III, the nucleic acid-protein assembly being laterally mobile relative to said support, such that the hybridization of a target nucleic acid to said nucleic acid IV or the change in conformation of said nucleic acid IV can be or is detected via the geometric reorganization of the nucleic acid-protein assembly.\",\n \"In a preferred case of the present invention, said protein III is chosen from the group consisting of proteins having a redox center and proteins having an anisotropic optical absorption property.\",\n \"The protein III will be described in greater detail later.\",\n \"The term “cell” is intended to mean a laterally delimited subset of the surface, which is characterized by a structural homogeneity, and many identical or variant copies of which can be reproduced on the inorganic support.\",\n \"A cell can in particular be used for detecting a specific type of nucleic acid (as will be seen later), different from the nucleic acid detected by using another cell.\",\n \"It is understood that the present invention also relates to a support displaying a plurality of cells according to the invention.\",\n \"In the present application, the term “floater” may be used to denote the assembly consisting of the protein which is mobile relative to the support, attached to the oligonucleotide.\",\n \"The oligonucleotide linked to the floater may be referred to as “primer” in the present application.\",\n \"The following abbreviations may also be used in the present application: DMPC: dimyristoylphosphatidylchloline DOGS: 1,2-dioleoyl-sn-glycero-3-[(N-(5-amino-1-carboxy-pentyl)iminodiacetic acid)succinyl] DOPC: dioleoyl phosphatidylcholine DPDPB: 1,4-di-[3′-(2′-pyridyldithio)propionamido]butane DPPC: dipalmitoyl phosphatidylcholine dsDNA: double-stranded DNA DTT: dithiothreitol Hb5(His)4: human cytochrome b5-histidine 4 fusion protein Hb5(His)4mut24: cysteine (position 24) mutant of Hb5(His)4 MIAC: metal ion affinity chromatography OG: octyl glucopyranoside OM: octadecyl mercaptan SPR: surface plasmon resonance RU: response unit SAM: self assemblage monolayer ssDNA: single-stranded DNA It is advantageous for the oligonucleotide not to be directly in contact with the support, thereby allowing an improved display with respect to the target nucleic acids or other elements which interact and modify the conformation of the primer.\",\n \"In a particular case, the protein-nucleic acid assembly is mobile relative to the support due to the fact that the protein is itself mobile in a lipid layer which is itself attached to the support.\",\n \"A lipid bilayer is preferably used.\",\n \"Briefly, this assembly consists of a lipid membrane reconstituted on a flat surface which is inorganic in nature.\",\n \"This is then referred to as a supported lipid membrane.\",\n \"Such a model has properties which are close to biological membranes in terms of compactness of the elements making it up, of compartmentalization, and of fluidity of the elements making up these models and also of most of the molecules incorporated or interacting with this model (Heyse et al., 1998, Marchal et al., 1998).\",\n \"This property of membrane dynamics is the reason for the lateral mobility of the proposed structures and for the reorganization allowing signal transduction and domain formation.\",\n \"In addition, it is possible to modulate as desired the molecular composition of this lipid structure in order to modify its intrinsic properties or to introduce new potentialities.\",\n \"Thus, in a preferred embodiment, the present invention relates to a cell for displaying a nucleic acid, comprising: a support which is substantially flat on an atomic scale, to which is attached a hydrophobic monolayer I, on which is present a monolayer II comprising phospholipids, a bridging molecule having a hydrophobic end which can interact with the monolayer II, and an end which is chemically functionalized so as to form a stable bond with a protein III, a protein III having an unequivocal three-dimensional structure, said protein III being laterally mobile in said layer II, a nucleic acid IV unequivocally attached to said protein III, preferably via a molecular arm.\",\n \"The fact that the protein is mobile in the layer II (lipid sheet) ensures lateral dynamics, i.e.\",\n \"the possibility of the floater+primer assembly diffusing freely in two dimensions in the absence of any additional geometric constraint which would be imposed by interaction of the primer with other nucleic acids.\",\n \"The membrane fluidity allows the reorganization and the contact of molecules essential for the self-assembly into superstructures.\",\n \"In addition, in some embodiments, the supramolecular assembly thus produced is characterized by a strong modularity: each step for producing or for destructuring this assembly may be carried out irreversibly by the controlled action of the solutes present in an aqueous phase or by the action of thermal conditions or of an electrostatic environment, etc.\",\n \"The grafting of a single-stranded nucleic acid onto the protein has been carried out so as to use the very particular properties of these biostructures.\",\n \"In this way, the oligonucleotides are grafted onto blocks which are protein in nature and which are floating on the upper lipid monolayer of the membrane model.\",\n \"This arrangement offers the following advantages: Lateral mobility of the floating protein anchor and, consequently, of the grafted ssDNA.\",\n \"Suppression of the nonspecific interactions of the ssDNAs on the flat support due to an appropriate lipid covering.\",\n \"Control of the density of the ssDNAs present at the surface of the biosensor (great modularity).\",\n \"In some embodiments, reversibility of the various supramolecular interactions via aqueous treatment procedures (allowing regeneration of the biosensor).\",\n \"The terms “nucleic acid”, “nucleic acid sequence”, “polynucleotide”, “oligonucleotide”, “polynucleotide sequence” or “nucleotide sequence”, terms which may be used indifferently in the present description, are intended to denote a precise series of natural or synthetic nucleotides defining a fragment or a region of a nucleic acid and which can correspond equally to a single-stranded, double-stranded or triplex DNA or RNA.\",\n \"Thus, the nucleic acid sequences according to the invention also encompass PNAs (peptide nucleic acids), or the like.\",\n \"The bonds between the various bases may be conventional phosphodiester bonds, or modified bonds such as phosphorothioates or methylphosphonates.\",\n \"The oligonucleotides may also be chimeric, i.e.\",\n \"they may have different bonds and/or bases.\",\n \"The term thus encompasses any variant of a nucleic acid sequence which exhibits a property of recognition by hybridization.\",\n \"Preferably, the nucleic acid linked to the protein in the cell according to the invention is a single-stranded nucleic acid.\",\n \"Its size is not a determinant parameter according to the invention, and it can be readily chosen using the general knowledge of those skilled in the art.\",\n \"In a preferred embodiment, the nucleic acid is attached to said protein via a reversible bond.\",\n \"This reversible bond may, moreover, be covalent.\",\n \"Thus, it is possible to attach the nucleic acid to the protein such that disulfide bonds are involved, which bonds can, however, be reversed in the presence of appropriate agents, or under particular conditions.\",\n \"Thus, the reversible bond can, in this particular case, be considered to be a bond which can be readily broken.\",\n \"In one embodiment, the nucleic acid is attached to said protein via a covalent bond.\",\n \"Such a bond is known to those skilled in the art and can in particular be produced using modified bases in the oligonucleotide allowing coupling to a chemical arm which is functionalized so that it can be coupled specifically to the protein of the floater.\",\n \"Moreover, there can be two types of grafting: a closely related grafting can be carried out so as to place the nucleic acid molecule and the protein tightly next to one another, or grafting via a molecular spacer arm can be carried out so as to provide greater conformational freedom of the DNA with respect to the floater.\",\n \"A spacer arm attached in an unequivocal manner to said protein is therefore preferably used.\",\n \"In fact, as will be developed later, it is advantageous for the geometry of the floater to be completely defined.\",\n \"Thus, it is preferable for the spacer arm to be attached to the protein at a precise site of said protein.\",\n \"In the embodiment in which the nucleic acid and the protein are placed next to one another, said spacer arm can then be a metal complex such as cis-platinum, trans-platinum, europium or another transition metal involving ligands originating from the very structure of the nucleic acid and one or more ligands originating from the very structure of the protein III of the floater.\",\n \"When the spacer arm is longer, allowing greater flexibility of the nucleic acid, a spacer arm is preferably chosen which attaches a thiol- or amino-function introduced onto the nucleic acid, in particular via a modified base (it being possible for the base chosen to be any base, i.e.\",\n \"it is possible to choose a base in the 3′ position, in the 5′ position or within the oligonucleotide, this alternative possibly having certain advantages, in particular of providing the oligonucleotide with a greater degree of freedom), to another functionality present, preferably uniquely present, on the protein.\",\n \"This second function is preferably also a thiol function, carried for example by a cysteine.\",\n \"However, other types of grafting of a nucleic acid to a protein exist.\",\n \"They can in particular be grafted onto an arginine in the protein.\",\n \"It is, however, preferable, in order to respect a principle of the invention, for the coupling of the nucleic acid to the protein to be specific, completely defined and unequivocal in molecular terms.\",\n \"Thus, any type of random grafting of the nucleic acid onto the protein which may create an ambiguity or a heterogeneity in the structure of the floaters according to the invention is generally excluded.\",\n \"Thus, preferably, the nucleic acid IV is attached to said protein III via a compound chosen from: a spacer arm connecting two identical or different, chemically reactive residues of the nucleic acid IV and of the protein III, chosen from the group consisting of thiols, amines, amides and arginines; a metal complex such as cis-platinum, trans-platinum, europium, or a complex of nickel, of copper or of ruthenium coordinated, firstly, with an amino residue or a group of amino residues of said protein III and, secondly, with one or more natural or modified bases of the nucleic acid IV, a covalent or noncovalent protein complex involving the nucleic acid IV.\",\n \"The protein in the cell according to the present invention has a geometry which can be detected in an unequivocal manner, i.e.\",\n \"a stable and defined spatial organization.\",\n \"This means in particular that its conformation in the cell according to the present invention is identical whatever the protein observed, when the cell according to the invention displays several identical floaters.\",\n \"This also means that bringing the cell into contact with outside nucleic acids capable of interacting with the primers in an unequivocal manner will lead to a highly repetitive organization of the floaters, in spatial terms, within the cell.\",\n \"In particular, the external nucleic acids may bring about the association of floaters in organized groups and these groups of floaters may, by virtue of their lateral mobility, interact with one another noncovalently so as to form, within each cell, a network of floaters which is highly organized in two-dimensional terms.\",\n \"The properties (physical, electrical, optical, quantum properties) of this highly organized network will then differ from the dynamic properties of disorganized floaters within the cell in the absence of any interaction with the external nucleic acid.\",\n \"Preferably, the protein of the floater will be chosen such that the variations in spatial organization of the floaters relative to one another lead to a signal which can be detected by any method, in particular an electrical, optical or resonance method.\",\n \"Proteins having this property will preferably be chosen from the group consisting of proteins having a redox center and/or an anisotropic optical absorption property and/or an intrinsic fluorescence property.\",\n \"Mention is in particular made of cytochromes, and more preferably human cytochrome b5, yeast cytochrome b5 or cytochrome b5 of any other origin, flavodoxins, ferridoxins, azurins and other blue copper proteins, and multiheme proteins of the bacterial cytochrome c class, which have both a redox center and anisotropic optical absorption properties.\",\n \"Among the fluorescent proteins, mention will preferably be made of green fluorescent protein (GFP) and analogs thereof obtained by site-directed mutagenesis or directed evolution.\",\n \"In a preferred case of implementation, said protein is water soluble.\",\n \"It may also have undergone modifications, some of which are indicated below, provided that these modifications result in a protein with a stable and defined three-dimensional structure satisfying the criteria as defined according to the present invention.\",\n \"Thus, in a preferred embodiment, said protein has undergone a modification which has allowed the introduction of a unique site into said protein, in order to effect said bond with said nucleic acid.\",\n \"The fact that a unique site is introduced into the protein according to the invention, in order to graft the nucleic acid, thus makes it possible to strongly promote the unicity, the specificity and the unequivocal definition of the protein-nucleic acid bond.\",\n \"In a particular case, said modification consists of a mutation of said protein so as to leave remaining or to introduce just one predetermined unique amino acid.\",\n \"The mutation can be introduced by mutation of the nucleic acid encoding the protein; this is the simplest method which will be favored by those skilled in the art.\",\n \"The unique site for grafting the nucleic acid will then be this predetermined unique amino acid.\",\n \"It is in fact within the scope of those skilled in the art to determine, according to the sequence of the protein which has been chosen in order to implement the invention, the amino acids to be modified so as to leave remaining just one of them, or so as to introduce one which is not present in the native protein.\",\n \"Thus, and in order to illustrate this fact, when the protein which is used is a cytochrome b5, it is possible to introduce a cysteine into the protein, since this amino acid is absent from the native protein.\",\n \"It is, moreover, always very advantageous to modify the protein such that it has only one cysteine, or at least such that it has a unique free cysteine (not involved in a disulfide bond with another cysteine).\",\n \"It is also preferable, for correct implementation of the method according to the invention, for the modification in the protein to be such that this facilitates the grafting of the nucleic acid, i.e.\",\n \"such that the amino acid used for said grafting is located at the periphery (at the surface) of said protein.\",\n \"It is within the scope of those skilled in the art to determine, as a function of the protein chosen, the best position for introducing the modification into the protein.\",\n \"In cytochrome b5, introduction of the cysteine by mutation of serine 24 has thus been exemplified.\",\n \"However, alternative positions could easily have been chosen.\",\n \"Thus, according to the three-dimensional structure of a cytochrome b5 of a mammal other than human (for example rat or bovine), it is entirely possible to select the amino acids to be modified so as to introduce just one cysteine into the sequence of the protein.\",\n \"For example, the representation in three dimensions will make it possible to target the most peripheral amino acids of the protein, i.e.\",\n \"those which promote better reactivity with the modified nucleic acid.\",\n \"In addition, it may be of crucial advantage to link the ssDNA in the region of the redox center of the protein: a mutation of glycine 47 would make it possible to obtain such a connection.\",\n \"One of the important characteristics of the present invention is that the protein III is mobile relative to the layer II of the cell according to the invention.\",\n \"This also ensures the mobility of the nucleic acid IV attached to the protein III in the cell, relative to the support according to the invention.\",\n \"The mobility of the protein in the layer II can be obtained in various ways, in particular when said protein has a hydrophobic tail which allows it to be anchored in the layer II.\",\n \"The sequences of hydrophobic tails are known in the literature, and it is within the scope of those skilled in the art to choose such a sequence and to modify the selected protein by genetic engineering in order to introduce such an anchoring sequence.\",\n \"It is then advantageous to choose a hydrophobic tail which is short, so that it becomes anchored only in the layer II, without penetrating into the layer I.\",\n \"This makes it possible in particular to improve the mobility of the protein in the layer II, by reducing the “rubbing” phenomena which may be observed if the hydrophobic tail is too large.\",\n \"The optimization of these elements remains within the scope of those skilled in the art.\",\n \"In a preferred case, however, said protein is attached to a phospholipid of the layer II and, in a particular case, to an artificial phospholipid preferably introduced in a small proportion (i.e.\",\n \"in a proportion less than 10%, preferably less than 5%, more preferably approximately 2%).\",\n \"As a result, the mobility of the protein relative to the layer II is then similar to the mobility of the phospholipids in said layer.\",\n \"Such a mobility is generally greater than when the protein has a hydrophobic tail.\",\n \"This makes it possible to improve the levels of performance of the cell according to the invention, as will be seen.\",\n \"Those skilled in the art have several methods available to them for producing a protein-phospholipid bond.\",\n \"Mention may in particular be made, among covalent bonds, of acylation, farnesylation, the use of a GPI (glycosylphosphatidylinositol) anchoring sequence or the incorporation of a phospholipid or of a fatty acid into a specific residue of the protein III by any artificial method.\",\n \"It may, however, be preferable to have a reversible bond, i.e.\",\n \"a bond which can be broken in the presence of reagents or under particular conditions, but which is stable under the usual working conditions or in the presence of the usual working reagents (hybridization of nucleic acids, study of binding between biological elements, etc.).\",\n \"In general, those skilled in the art can use all the known methods for attaching a protein to a liposome, for example, since liposomes generally consist of phospholipids.\",\n \"Thus, it is in particular possible to use an arm capable of linking a metal chelate on one side and an amino acid on the other, using, in the layer II, a modified phospholipid, said phospholipid having a chelate-binding zone.\",\n \"Thus, a preferred phospholipid according to the invention is DOGS, or 1,2-dioleoyl-sn-glycero-3-[(N-(5-amino-1-carboxypentyl)iminodiacetic acid)succinyl].\",\n \"This phospholipid is a synthetic phospholipid possessing an iminodiacetate residue which complexes a nickel divalent cation.\",\n \"In order to promote the mobility of the protein in the layer II, it is advantageous to choose the phospholipids of said layer II so that they have hydrophobic tails chosen such that said layer is not in the crystalline state, but in the fluid state at the temperature for use.\",\n \"These properties of fluidity of a phospholipid layer are known to those skilled in the art, and there are in particular diagrams for choosing the chain length and the unsaturation in order to obtain fluidity at the temperature for use (Gulik-Krzywicki et al., 1967).\",\n \"This temperature for use obviously depends on the use for which the sensor is intended.\",\n \"Thus, when hybridization is sought, the temperature for use may be different from that used when the binding of a protein to a nucleic acid is studied.\",\n \"Furthermore, transition between fluid and gel states may be strategic in the context of a detection technique based on super-structure imaging, such as atomic force microscopy (AFM).\",\n \"Those skilled in the art will seek the phospholipids most suitable for the layer II as a function of the temperature for use.\",\n \"As a general rule, and in order to simplify and improve the potentialities of the biosensor, fluidity of the layer II at ambient temperature is sought.\",\n \"Thus, phospholipids having hydrophobic tails which are greater than or equal to 14 carbon atoms in chain length are preferably chosen, said chain optionally having unsaturations, such that said layer II is in the fluid state at the temperature for use.\",\n \"It is advantageous to note, for example, that C14 elements are generally fluid at a temperature of greater than 23° C., whereas C16 elements are fluid only from 37° C., and C18 elements above 50-55° C. However, the use of C18 tails which have unsaturations makes it possible to decrease the phase transition (crystalline state-fluid state) temperature.\",\n \"Nevertheless, for some applications or methods of detection, it may be preferred for the layer II not to be fluid at ambient temperature, but only at higher temperatures.\",\n \"In particular, this method will be preferred if it is desired to “memorize” the organizational state of the floaters, using a decrease in temperature to fix the structure after an organization phase at higher temperature.\",\n \"This will in particular be the case if there is a risk of the detection technique (near-field microscopy for example) or the conditions for analysis (dehydration for example) altering the structure.\",\n \"The nonmodified phospholipids of the layer II have no particular characteristics besides the mobility properties already described; however, it is preferable for them to have polar heads, chosen from the group consisting of neutral polar heads carrying no charges, polar heads which are neutral overall but which carry opposite charges (zwitterionic polar heads), or polar heads carrying an overall negative charge.\",\n \"The choice of the charge carried by the phospholipids may prove to be advantageous, in particular for reducing the phenomena of nonspecific absorption of the test nucleic acids on the layer, which are decreased when the charge of the phospholipid heads is negative.\",\n \"The use of neutral heads (either uncharged or carrying several opposite charges) is also envisioned.\",\n \"The layer II is assembled on the layer I conventionally, via the interactions which exist between two hydrophobic elements in an aqueous medium.\",\n \"Reference may therefore be made to a self-assemblage layer.\",\n \"However, in certain cases, a suitable catalyst, called fusogenic agent, will be used to facilitate this fusion.\",\n \"Thus, such agents, such as Ca++ cations, the ethylene glycol polymer (PEG) or detergents, will be used according to protocols known to those skilled in the art.\",\n \"As regards the layer I, it comprises hydrophobic elements with a chain length of approximately 2 to 2.5 nm, for example of between 12 and 20 carbon atoms, preferably between 14 and 18 carbon atoms.\",\n \"It is in fact advisable to have a layer I which is particularly homogeneous.\",\n \"Moreover, the chain length for the elements of the layer I is preferably sufficiently long so that the coming together of the two layers I and II forms a true bilayer.\",\n \"The elements of the layer I are linear or branched and optionally carry unsaturations.\",\n \"It is important for this layer I to be homogeneous, since it serves as a support for depositing the layer II.\",\n \"The layer I is attached to the support of the cell according to the invention covalently, or via strong noncovalent bonds.\",\n \"This method of bonding to the support is not of great importance for the implementation of the invention.\",\n \"Thus, in a particular embodiment, said layer I comprises phospholipids linked to said support via their polar heads.\",\n \"In another embodiment, said layer I comprises hydrophobic elements linked to said support via a covalent bond, for example an alkyl-thiol bond or an alkyl-siloxane bond, depending on the nature of the support.\",\n \"It may be advantageous to have a support-layer I bond which is covalent, for facilitated re-use of the cell according to the invention.\",\n \"In fact, when the nucleic acid is linked to the protein and the protein is linked to the layer II by reversible bonds, the cell can be readily regenerated by the action of elements (or conditions) which make it possible to break the nucleic acid-protein and protein-layer II bonds and to eliminate the layer II by the action of an appropriate detergent.\",\n \"It is thus readily seen that a nonreversible covalent bond between the layer I and the support makes it possible to reconstruct a cell.\",\n \"It is obvious that if the layer I is linked to the support via a reversible bond, it is possible to recommence preparing the cell, but with an additional step.\",\n \"The re-use of the support can prove to be very advantageous insofar as said support must be virtually flat at the atomic level and, moreover, can include all or part of the system for detecting the state of the cells, which can pose certain manufacturing restrictions and increase the production costs for the cell according to the invention.\",\n \"Thus, in a preferred embodiment, the flatness of said support is such that the difference in height between two zones less than 100 nm apart is less than or equal to 10 nm, preferably less than or equal to 3 nm, more preferably less than or equal to 2 nm.\",\n \"Various materials may be chosen for producing the base of the cell according to the invention, in particular a material chosen from the group consisting of glass covered with a layer of gold, of cleaved mica, of silicon or of any other monocrystalline material.\",\n \"Monocrystalline silicon is a very advantageous material insofar as it is effectively flat on an atomic scale, and it can readily be subjected to microetching and varied chemical modifications, which makes it possible to produce a support having a plurality of cells according to the invention.\",\n \"Moreover, silicon has conducting or optical properties which may prove to be very advantageous as regards the detection of biological events taking place on the cell according to the invention.\",\n \"As will be developed, the cell according to the invention may comprise several identical or different proteins linked to identical or different nucleic acids.\",\n \"When different primers are used, a target nucleic acid complementary to two primers will be capable of binding the corresponding two floaters, which will accentuate its interaction with the support and will prime a change in spatial organization of the floaters within the cell.\",\n \"Thus, when the nucleic acid IV-protein III and protein III-layer II bonds are reversible, it is possible, via agents which compete with the floater-support interaction (histidine, imidazole, EDTA), to readily differentiate the degrees of hybridization.\",\n \"Moreover, the aspect of two-dimensional reorganization of the floater and primer nano-objects on the flat membrane surface, obtained through the fluidity of the layer II, falls within the area of the self-organization of biological molecules and of the physicochemical phenomena which ensue from these collective movements.\",\n \"Thus, using a random distribution of the protein-nucleic acid blocks, it is clear that the action of an effector (for example a target molecule) will engender a radical restructuring in two dimensions.\",\n \"When use is in particular made of a cell in which two (or more) different primers are present, the presence in the target sample of a nucleic acid which can hybridize to these various primers and which allows said blocks to become bridged should lead to a process of elongation (one-dimensional organization) at the surface of the cell.\",\n \"There is therefore a change, and this is one of the major advantages of the invention, from a more or less disorganized system (random, or with an unstable equilibrium) to a highly ordered system, under the effect of the presence of the target nucleic acid in the sample tested.\",\n \"Moreover, this organization is extremely sensitive to the geometric conformation of the nucleic acid and therefore to any factor (mismatching, protein binding, chemical agent) capable of interacting with the conformation or the structure of the nucleic acid.\",\n \"Similarly, when the binding of any protein or of any ligand to a (single-stranded or double-stranded) nucleic acid linked to the protein anchored in the layer II is studied, or when the binding of an effector molecule on a protein (of the transcription factor type) linked to a nucleic acid, on a cell according to the invention, is studied, a modification of the structure (for example curvature of the nucleic acid) and of the equilibrium of the system are then observed (due to the introduction of these elements), which modifications can then be readily observed by various methods capable of detecting the organizational change in the cell, and in particular via an optical (absorbance, fluorescence), electrical, electron-based, surface plasmon resonance, energy transfer, radio-labeling, diffraction (optical, electrical, X-ray, neutron diffraction) or microscopy (direct optical or fluorescence microscopy, electron microscopy, near-field microscopy, atomic force microscopy) system.\",\n \"This disorder-order phase change observed when an event occurs on an atomic scale can only be readily detected when the protein on the cell according to the invention has what has been called “an unequivocally detectable geometry” and when the nucleic acid is attached thereto unequivocally.\",\n \"The principle of the invention is therefore the mobility of the protein anchored in the layer II, allowing a reorganization of the system, and the fact that the protein-nucleic acid unit is unequivocal and non-random, which makes it possible to detect said reorganization.\",\n \"It is also envisioned that the presence of a single target nucleic acid molecule in the sample tested will make it possible to induce a reorganization which may then be detected.\",\n \"An example of the implementation of such a method would be to pre-organize the cell with a nucleic acid of the same nature as that to be detected, but which, firstly, has a lower affinity for the primer than the nucleic acid to be tested (modified base, mismatch) and, secondly, has a geometry (size, curvature) which is different from the nucleic acid to be tested.\",\n \"The exchange of a single nucleic acid (external competitor molecule) within a pre-organized cell will then lead to a local structural defect.\",\n \"It is known by those skilled in the art that a single defect within a “crystalline” network destabilizes the entire network.\",\n \"If the cell size is chosen in an appropriate manner, it is reasonable to hope that this single event will change the stability of the organization of the cell.\",\n \"Such a change in stability can be tested by gradually applying an outside (chemical, temperature, electric potential, etc.)\",\n \"factor which destabilizes the cell as far as disorganization.\",\n \"This principle opens up the possibility of detecting single molecular events and corresponds to the theoretical maximum sensitivity of a detector.\",\n \"This sensitivity is an enormous advantage compared to the other existing widely used systems, which require the presence of a large number of nucleic acid molecules in the sample in order to be detected.\",\n \"Moreover, such a detector associated with a receptor protein chosen in an appropriate manner in order to see its interaction with the modulated DNA can be the basis of a molecular detector for an infinite variety of molecules of interest (in particular proteins, medicinal products, pollutants).\",\n \"It is also important to note that the hybridization as such is not detected, but rather the changes induced in the cell according to the invention.\",\n \"Thus, this implies that the cell according to the invention (or the support displaying a plurality of cells) can be used for detecting a target in a sample without it being necessary to label or to excessively handle said sample.\",\n \"This is also a considerable advantage, since it is thus possible to detect the presence of mRNA without having to first convert it into cDNA.\",\n \"It should be noted that the geometric conformation of ssDNAs and dsDNAs is so different that it results in very distinct physicochemical properties.\",\n \"Thus, after hybridization, it is in particular possible to detect this effect by a change in the electronic conductivity of the cell according to the invention.\",\n \"This is particularly easy when the protein according to the invention also has a redox center.\",\n \"It is also possible to detect the hybridization by surface plasmon resonance, or by an optical method, which is facilitated by the presence of a protein exhibiting an anisotropic optical absorption.\",\n \"In order to carry out the simultaneous detection of several targets, the invention therefore also relates to a support displaying a plurality of cells according to the invention.\",\n \"The various cells may be separated from one another by microetching on the support, or by the presence, between two cells, of a hydrophobic layer consisting of elements with a chain length different from (preferably less than) the chain length for the elements of the layer I, for example less than or equal to 5 carbon atoms (“differential patterning” Cheng et al., 2000).\",\n \"These elements having a low-carbon-number chain can be deposited onto the support using the masking techniques common in microelectronics.\",\n \"The support according to the invention is advantageously completed by the setting up of a system for measuring the current, the impedance or the potential, making it possible to detect the events occurring on each one of the cells.\",\n \"Another preferred alternative is that the support comprises a surface alternating conducting and reflecting zones and nonconducting and transparent zones.\",\n \"Such a network comprising transparent zones clearly smaller in size than the wavelength of visible light has been described as having a function of light transmission according to wavelength in the form of a “comb” (Ebbesen et al., 1998).\",\n \"The optical transmission properties of such a structure should be altered by the state of the neighboring cells, which provides a possible reading means.\",\n \"It can also be envisioned that the support according to the invention comprises, between each cell, an integrated optical device (laser-photodiode couple for example) capable of measuring the absorbance of a cell in two cross directions and of deducing therefrom its state of organization (anisotropy) or of disorganization (isotropy).\",\n \"It is also possible to preferably envision reading the organizational state of each cell via a laser optical scanning device sensitive to the change in light polarization.\",\n \"Such a device capable of reading cells which are submicron in size is commonly used by the general public in minidisc readers and rewritable CD-Rom readers.\",\n \"In such a case, the variation in polarization of the reflected light normally provided in magneto-optical discs by a reorganization of the crystalline network of a rare earth salt (europium, and other compounds of the same transition series of the Mendeleiev table) by a magnetic field and laser heating, would, in the system described in the invention, be provided by the variation in spatial organization, in the cell, of the protein or of its cofactor, which are objects which are optically active in terms of polarization, ellipticity (differential absorption of polarized light) and optical absorption.\",\n \"The invention also relates to a method for identifying the presence of a test nucleic acid in a sample, comprising the steps of: a) bringing said sample into contact with a cell according to the invention, under conditions which allow the hybridization of said test nucleic acid to a nucleic acid attached to a protein of said cell, b) detecting the hybridization of said test nucleic acid to said nucleic acid attached to said protein.\",\n \"The detection can be carried out by any means mentioned above.\",\n \"If the test sample has been labeled, the detection can be carried out as a function of said labeling.\",\n \"However, it may be preferred to detect the presence of the target nucleic acid by a change in conformation of the system.\",\n \"Thus, the cell according to the invention allows the detection without there having been any need to label the test sample.\",\n \"The invention also relates to a method for identifying the binding of a protein to a nucleic acid, and/or its activity on the conformation of said nucleic acid, comprising the steps of: a) bringing said protein into contact with a nucleic acid attached to a protein, in a cell according to the invention, b) detecting the binding of said protein to said nucleic acid, and/or its activity on the conformation of said nucleic acid.\",\n \"Said nucleic acid may be single-stranded, double-stranded or triplex, when a nucleic acid has already been attached to one of the nucleic acids (primers) of the cell.\",\n \"It is clear that the choice of certain bases for the primer can lead to the formation of a triplex nucleic acid, said formation then being considered, for the purpose of the invention, as a hybridization.\",\n \"The detection can also be carried out in several ways, and in particular by a change in conformation of the system.\",\n \"The invention also relates to a method for identifying the binding of a ligand to a protein attached to a nucleic acid attached to a protein in a cell according to the invention, comprising the steps of: a) bringing said ligand into contact with said protein, b) detecting the binding of said ligand to said protein.\",\n \"Among the ligands and proteins envisioned, studies are in particular carried out in order to identify effector molecules which bind to nuclear receptors or transcription factors.\",\n \"The binding may be direct or indirect, and may involve, for example, the binding of the ligand to a receptor which will then attach to a protein or a protein complex already associated with the DNA of the cell.\",\n \"The binding of the ligand to the protein can be observed in particular by a change in conformation of the system.\",\n \"In general, the invention relates to a method for identifying the binding of a compound to a nucleic acid, and/or its activity on the conformation of said nucleic acid, comprising the steps of: a) bringing said compound into contact with a nucleic acid attached to a protein, in a cell according to the invention, b) detecting the binding of said compound to said nucleic acid, and/or its activity on the conformation of said nucleic acid.\",\n \"As above, said nucleic acid may be single-stranded, double-stranded or triplex, when a nucleic acid has already been attached to one of the nucleic acids of the cell.\",\n \"The binding is in particular detected by a change in conformation of the system, as was explained above.\",\n \"The detection of the geometry or of the disturbance of the geometry of the Floater-Primer-Target assemblies can advantageously be carried out through the contribution of atomic force microscopy technology in a liquid environment.\",\n \"Thus, the potential applications of this type of characterization relate to studying the impact of the interaction of any molecules or macromolecules with nucleic acids (regulation, transcription factors, drugs, medicinal products, in particular for chemotherapy, etc.).\",\n \"In a particular case, the device according to the invention offers a molecular and supramolecular proximity involving a lipid membrane, redox proteins and nucleic acids.\",\n \"This assembly of electro-active and optically active biomolecules within microsystems derived from microelectronics may potentially prove to have a great impact in electron transduction (conductors, semi-conductors, biomolecular circuits) and nonlinear optical transduction (anisotropic) fields.\",\n \"In fact, double-stranded DNA, unlike ssDNA, has properties of electron transfer according to a process of electronic conduction by overlap of the Pi orbitals.\",\n \"In addition, the hybridization reaction between two ssDNA molecules is very specific.\",\n \"The current systems developed in the context of electronic components on a nanometer scale come up against various problems, including: the interconnection of the nano-objects with one another, the connection of the nanostructures to macroscopic electrodes.\",\n \"The use of processes of molecular recognition and of self-assembly of molecules within supramolecular structures, as envisioned within the cell according to the present invention, might make it possible to circumvent these difficulties.\",\n \"In fact, the cell according to the invention exhibits an original biomolecular architecture which falls within the field of nano-bioengineering, i.e.\",\n \"of the development of novel types of nanostructures having properties of: strong modularity, high degree of organization on a molecular scale, vectorial electron transfers, coupling with chemical messengers.\",\n \"DNA has the mechanical molecular recognition properties for being integrated into microcircuits.\",\n \"However, the intrinsic electronic properties of this macromolecule appear to be insufficient to be used directly (Braun et al., 1998).\",\n \"One of the most promising areas of research in the use of electrical charge transport potential along the DNA duplex is represented by the use of electrocatalytic processes (Boon et al., 2000).\",\n \"Such a signal transduction device can absolutely be integrated into the device according to the invention through the use of additional redox molecules.\",\n \"However, in a preferred device according to the invention, the presence of floaters composed of redox proteins having electroactive groups and/or redox potentials which are different and which can be modulated makes it possible to provide an electron environment which is dense in terms of overlap of electron orbitals.\",\n \"Depending on the periodicity and on the two-dimensional organization of these redox macromolecules (then considered to be relay centers), such a structure might give rise to electron doping of DNAs, to electron jumping between proteins or to energy transfer processes.\",\n \"Thus, a cell according to the invention may have methods of use other than the simple methods of detection which have been mentioned above.\",\n \"The invention also relates to a method for preparing a cell according to the invention, comprising the step of: introducing, on a support which is substantially flat on an atomic scale, a protein III having an unequivocal three-dimensional structure (stable and defined spatial organization), linked to a nucleic acid IV, such that the nucleic acid-protein assembly is laterally mobile relative to said support, in order for the hybridization of a target nucleic acid to said nucleic acid IV or the change in conformation of said nucleic acid IV to be detectable via the geometric reorganization of the nucleic acid-protein assembly.\",\n \"The invention also relates to a method for preparing a cell according to the invention, comprising the steps of: attaching a hydrophobic monolayer I to a support which is substantially flat on an atomic scale, assembling, on said monolayer I, a mono-layer II comprising phospholipids, introducing a protein III having an unequivocal three-dimensional structure, said protein III being laterally mobile in said layer II, via a bridging molecule having a hydrophobic end capable of interacting with the monolayer II, and an end which has been chemically functionalized so as to form a stable bond with a protein III, said protein III being attached in an-unequivocal manner to a nucleic acid IV, preferably via a molecular arm.\",\n \"DESCRIPTION OF THE FIGURES FIG.\",\n \"1: Construction of the hybrid bilayer and characterization by surface plasmon resonance analysis A: The signal recorded corresponds to the base line for the metal support covered with a dense monolayer of OM in the environment of the working buffer (conventional phosphate buffer, 50 mM, pH 7.5).\",\n \"B: The variation in signal corresponds to the injection into the fluid circuit of a DMPC-DOGS mixed vesicular solution produced in water.\",\n \"The increase in the signal corresponds to fusion kinetics for the vesicles on the support so as to obtain a compact lipid monolayer.\",\n \"Thus, after 20-40 min of injection, a return to the working buffer makes it possible to estimate the amount of lipids assembled at 1700±300 RU.\",\n \"This value, according to the various modes of calibration proposed by BIAcore, corresponds to that of a lipid half-membrane.\",\n \"C: A procedure for treating the membrane with a 20 mM sodium hydroxide solution makes it possible to remove the excess lipids adsorbed onto the structure.\",\n \"D: Control of the homogeneity of the lipid membrane by injection of water-soluble protein (for example cytochrome c, BSA, etc.).\",\n \"FIG.\",\n \"2: Demonstration of a specific association of Hb5 (His)4 on the hybrid bilayer functionalized with chelating nickel cations Characterization by SPR A: Three successive injections (represented by arrows) of the cytochrome b5 polyhistidine form are carried out in the region of the functionalized lipid bilayer.\",\n \"Conventionally, the protein solution, which has a concentration of 1 μM in 50 mM phosphate buffer, pH 7, is injected at a flow rate of 10 μl/min for 180 seconds.\",\n \"The various injections produce a gradual saturation of the surface.\",\n \"The maximum covering is of the order of 650±50 RU, i.e.\",\n \"a degree of surface covering of 7 pmol.cm−2.B: In order to show the great specificity of the interaction, injections of a 1 mg/ml solution of histidine in 50 mM phosphate buffer are carried out (for example: four injections marked with arrows).\",\n \"The injection of an agent which competes with the chelation between the polyhistidine segment of Hb5 (His)4 and the membrane.\",\n \"Each injection (flow rate 10 μl/min for 120 seconds) is accompanied by a decrease in signal, i.e.\",\n \"in the amount of proteins interacting with the membrane.\",\n \"Thus, complete regeneration of the structure is obtained after a contact time of a few minutes.\",\n \"FIG.\",\n \"3: Diagram of the membrane biosensor, construction/regeneration of the supramolecular assembly Construction: An inorganic surface (for example: gold) is functionalized with a dense monolayer of octadecyl mercaptan.\",\n \"This surface is suitable for the fusion of lipid vesicles (liposomes) until a dense monolayer of phospholipids is obtained above the support.\",\n \"The presence of 10% (mol/mol) of modified phospholipids allows the anchoring of cytochromes b5 so as to obtain a dense monolayer of proteins.\",\n \"Regeneration: Injections of agents which compete with the affinity via metal ions, such as a solution of histidine (1 mg/ml), make it possible to detach all the Hb5(His)4 interacting specifically with the support.\",\n \"Similarly, the lipid half-membrane reconstituted on the functionalized inorganic support can be regenerated using a solution of detergent such as octyl glucoside, or OG (40 mM in H2O).\",\n \"Thus, all of the constructing and destructuring phases can be carried out in a controlled manner using an aqueous solution which preserves the integrity of the biological molecules.\",\n \"FIG.\",\n \"4: Characterization of the b5-cis-Pt-ssDNA complex, SDS-polyacrylamide (15%) gel of Hb5(His)4 platinated and coupled with an oligonucleotide of sequence CTATCATTTGCTTACTATTC (SEQ ID No.\",\n \"13) FIG.\",\n \"4.A: Autoradiogram of a gel into which Hb5(His)4-cis-Pt has been injected; the 32P-radiolabeled oligonucleotide (well 2) and the b5-cis-Pt-ssDNA complex (well 3) make it possible to demonstrate the appearance of an additional radioactivity signal in well 3 compared to well 2.FIG.\",\n \"4.B: Use of a polyacrylamide gel (15%) under denaturing conditions (SDS).\",\n \"Cross-characterization of the b5-cis-Pt-ssDNA complex: said complex injected into lane 3 exhibits a band which makes it possible to confirm the existence of a population whose molecular weight is close to 20000 Daltons, i.e.\",\n \"the theoretical molecular weight.\",\n \"Well (1): b5 platinated with cis-Pt[(NH3)2(H2O)2]2+.\",\n \"Well (2): radiolabeled 20-mer DNA (DNA*).\",\n \"Well (3): platinated b5 linked to the 20-mer DNA* FIG.\",\n \"5: Spectral characterization of the complex Hb5(His)4-CTATCATTTGCTTACTATTC via cis-Pt[(NH3)2(H2O)2]2+bridging Two procedures for reconstituting the Hb5 (his)4-cis-Pt-ssDNA complex are characterized by spectrophotometry in a 240-600 nm window.\",\n \"The cytochrome b5 has an absorption maximum at 412 nm whereas the ssDNA has a maximum at 260 nm.\",\n \"The spectral profile of two samples and also the relative quantification of each species by virtue of the molar extinction coefficients make it possible to demonstrate: red spectrum: an Hb5(His)4/ssDNA molar fraction= 3/1 blue spectrum: an Hb5(His)4/ssDNA molar fraction= 1/1 The procedure for the characterization represented by the blue absorption spectrum makes it possible to obtain equimolar coupling.\",\n \"FIG.\",\n \"6: Determination of the melting temperatures of the 20-mer DNA and of the 20-mer DNA bridged with platinated Hb5(His)4 The melting temperatures were determined by spectral measurements of hyperchromicity after melting of the complementary strand at high temperature (80° C.).\",\n \"A decrease by steps of 1° C. makes it possible to follow the evolution of the dsDNA spectrally.\",\n \"The results of absorption as a function of temperature for the free ssDNA makes it possible to obtain a curve whose inflexion point corresponds to the melting temperature.\",\n \"The Tm for the ssDNA is evaluated at 45° C. The same characterization on the ssDNA coupled to Hb5(His)4 gives a modified profile, the Tm of which was estimated at 23° C. FIG.\",\n \"7: Separation of the protein species and of the cytochrome b5-ssDNA complex via a DPDPB bridge, by chromatography.\",\n \"Characterization by spectrophotometry at 240-450 nm The various populations derived from the incubation (4 H at 30° C.) of ssDNA-DPDPB with Hb5(His)4mut24 are loaded onto a DEAE column.\",\n \"The various species are then eluted via a 25 mM phosphate solution and an increasing NaCl gradient.\",\n \"The cytochrome b5 has an absorption maximum at 412 nm whereas the ssDNA has a maximum at 260 nm.\",\n \"The NaCl gradient makes it possible to gradually elute all the populations: [NaCl]<0.15 M: population 1 of Hb5(His)4mut24 0.15<[NaCl]<0.25 M: population 2 of Hb5(His)4mut24 [NaCl]=0.5 M: population 3 of Hb5(His)4mut24 FIG.\",\n \"8: Elution profile and supramolecular composition of the cytochrome b5-ssDNA complexes The molar quantification of the species isolated after separation on DEAE is represented in the form of a histogram: Populations 1 and 2 of Hb5(His)4mut24, characterized in FIG.\",\n \"7, show no ssDNA component measured at 260 nm.\",\n \"These two populations correspond to the monomeric form (which is not reacted during the incubation) and to the dimeric form (resulting from a cystine bridge between two proteins).\",\n \"Population 3 shows an equimolar Hb5(his)4mut24-ssDNA complex.\",\n \"FIG.\",\n \"9: Detection by surface plasmon resonance of the kinetics of association of an ssDNA complementary to the primer ssDNA interacting with the biosensor Starting from a “b5-W1” signal in the region of 250 RU, various injections of ssDNA were carried out, at a b5-ssDNA density equal to 1 pmol.cm−2: Thus, the ssDNA strand complementary to W1 was injected in the region of the biosensor (1 μl/min for 10 min) for two concentrations of NaCl.\",\n \"At a 150 mM concentration of NaCl, a hybridization signal is observed within a complexation range corresponding to 100% of the “b5-ssDNA” assemblies interacting with the membrane.\",\n \"The same oligonucleotide injected at a low concentration of NaCl (25 mM) shows very restricted hybridization with respect to the biosensor.\",\n \"FIG.\",\n \"10: Diagram of the assembly as a ˜DNA biosensor The arrow represents the flow of biomolecules of the test sample.\",\n \"FIG.\",\n \"11: Diagram of the assembly as nanostructures with dynamic and conformational properties A set of various floating blocks can be displayed at the surface of the biosensor, and the injection of an oligonucleotide which potentially hybridizes on a part of the floating blocks should allow inter-block bridging.\",\n \"The lateral mobility characteristic of the elements should promote this process until phenomena of supramolecular reorganization at the surface of the cell are obtained.\",\n \"The following examples relate to a particular case of the invention, but should not be considered as limiting the invention.\",\n \"EXAMPLES Example 1 Preparation of the Hybrid Bilayer 1.1) Functionalization of the Support The solid support used consists of a glass slide covered with a film of gold marketed by the company BIAcore® under the name chip J1.Such supports were also produced by the inventors.\",\n \"They consist of the use of polished glass cover slips (thickness 0.4 mm, 22*10 mm, Prolabo) which are successively subjected to spraying of 2.6 nm chromium and then spraying of 48 nm of gold under a vacuum of 2×106 mmHg.\",\n \"The metal interface is then covered by self-assembly of molecules of octadecyl mercaptan, or OM, from Aldrich.\",\n \"The OM solution is used at 1 mM in a 4/1 (v/v) ethanol/water mixture.\",\n \"After incubation for two hours, the slides are washed with chloroform, with methanol and then with water and are dried under a stream of nitrogen.\",\n \"The result is the reconstitution on the gold of a highly organized dense monolayer which confers on the support very strong hydrophobic properties.\",\n \"This first assembly has been characterized many times in the literature.\",\n \"Contact angle measurements make it possible to routinely verify the compactness of the monolayer thus reconstituted.\",\n \"1.2) Production of the Lipid Xesicles 1.2.1) Sonication/Extrusion Method The method used to prepare the liposomes is based on a sonication/extrusion double procedure.\",\n \"The dimyristoyl and/or dipalmitoyl phosphatidyl-choline phospholipids from Sigma, which are the main constituents of the vesicles, are solubilized in an organic solution of chloroform.\",\n \"A lipid film is then deposited in glass scintillation tubes by evaporation of the chloroform under a stream of nitrogen.\",\n \"This film is resuspended in an aqueous solution (of ultrapure or buffered water) at a final concentration of 1 mM.\",\n \"This mixture is then sonicated: in an ultrasound bath (Transsonic 310 from Fisher-Scientific) at maximum power for 20 min, and then with an ultrasonic disintegrator (Vibra-Cell from Bioblock) using 2 min pulses, with 50% active cycle for an outlet power of 4.The procedure is stopped when the solution has become translucent, i.e.\",\n \"conventionally after 6 to 10 min of cumulative sonication time.\",\n \"The final production step consists of extrusion through a polycarbonate membrane (Avestin, 19 mm in diameter and 50 nm or 100 nm in pore diameter) in a Liposofast® basic device, Avestin, Inc. 1.2.2) Vortex/Extrusion Method A second procedure is used when molecules are used which are sensitive to ultrasonic disturbances, such as the hydrocarbon chains of some phospholipids having polyunsaturations (for example: 1,2-dioleoyl-sn-glycero-3-[(N-(5-amino-1-carboxypentyl)iminodiacetic acid)succinyl] or DOGS, from Aventi Polar Lipids, and dioleoyl phosphatidylcholine, or DOPC, from Sigma).\",\n \"In this case, a procedure identical to 1.2.1, consisting of solubilization in CHCl3 and then construction of a lipid film on glass, is applied.\",\n \"The procedure then differs in that the phospholipids are resuspended in an aqueous solution at 1 mM via a step consisting of vortexing for 5 min at maximum rate.\",\n \"This step is followed by a phase of extrusion through a polycarbonate membrane with a 50 nm pore diameter, in a Liposofast® device from AVESTIN.\",\n \"1.3) Reconstitution of the Lipid Monolayer The hydrophobic support serving as matrix to generate the lipid monolayer is subjected to a treatment procedure as follows: in order to improve the wettability of the structure, an ethanol/water ( 1/1 vol) solution is injected onto the support and is followed by steps of rinsing with water and with buffered solutions.\",\n \"In order to clean the interface, a solution of detergent, octyl glucopyranoside from Sigma, at a concentration of 40 mM, is injected.\",\n \"Immediately following this, the lipid vesicles prepared are injected in contact with the functionalized metal surface.\",\n \"The vesicles fuse spontaneously on contact with the hydrophobic monolayer of OM, according to a liposome fusion process established by Kalb et al.\",\n \"(1992).\",\n \"After 30 to 60 min, this process leads to the formation of a lipid monolayer completely covering the OM monolayer.\",\n \"The molecular assembly thus obtained has a substrate which is metallic in nature, covalently covered with a first alkylated sheet on which is assembled a fluid and dynamic sheet of amphiphilic molecules.\",\n \"The characterization experiments use the BIAcore® technology via BIAcore 1000 and BIAcore X devices.\",\n \"The liposome fusion results are obtained in the form of a sensorgram (FIG.\",\n \"1).\",\n \"On injection of the vesicles, a phase of association with the support is observed, producing a plateau after a contact time of 20-30 [lacuna].\",\n \"On stopping the injection, and therefore on returning to the working buffer, a first amount of lipid material is evaluated.\",\n \"A procedure consisting of cleaning the surface with 20 mM sodium hydroxide makes it possible to remove all the vesicles incompletely fused at the surface of the biosensor.\",\n \"Conventionally, the signal obtained is between 1400 and 2000 RU, which is in accordance with the results available in the literature and the methods of calibration provided by the manufacturer.\",\n \"An injection of soluble proteins in the region of this supported membrane makes it possible to validate the absence of a defect in the upper lipid monolayer.\",\n \"Introduction into the liposomes of synthetic DOGS phospholipids in a proportion of 10% mol/mol confers on this monolayer protein-HisTag sensor complexation properties.\",\n \"Example 2 Construction, Expression and Purification of the Fusion Proteins 2.1) The Water-Soluble Proteins with a Histidine Tag The initial protein used in the context of this example is a membrane-bound heme protein: human or yeast cytochrome b5.It is a bitopic protein having a large globular water-soluble domain containing the heme and a small hydrophobic domain allowing anchoring to microsomal membranes.\",\n \"All the nucleotide and amino acid sequences are included in the sequence listing.\",\n \"The amino acid sequence of the human and yeast forms are referred to as follows: Amino Acid Sequence of Membrane-Bound Human Cytochrome b5 (SEQ ID No.\",\n \"1) 1 MLAEQSDEAV KYYTLEEIQK HNHSKSTWLI LHHKVYDLTK FLEEHPGGEE VLREQAGGDA 61 TENFEDVGHS TDAREMSKTF IIGELHPDDR PKLNKPPETL ITTIDSSSSW WTNWVIPAIS 121 AVAVALMYRL YMAED Amino Acid Sequence of Membrane-Bound Yeast Cytochrome b5 (SEQ ID No.\",\n \"2) 1 MPKVYSYQEV AEHNGPQNFW IIIDDKVYDV SQFKDEHPGG DEIIMDLGGQ DATESFVDIG 61 HSDEALRLLK GLYIGDVDKT SERVSVEKVS TSENQSKGSG TLVVILAILM LGVAYYLLNE The first experiment to modify the sequences of cytochrome b5 consisted in the removal, on the carboxy-terminal side, of the amino acids involved in the membrane insertion, so as to replace them with four histidines.\",\n \"This chimeric or fusion protein is obtained by cloning the nucleotide sequence by polymerase chain reaction (PCR).\",\n \"The cloning kit used is TOPO TA cloning® from Invitrogen.\",\n \"The coding DNA sequence for the chimeric protein is introduced into a pCR®2.1-TOPO® vector.\",\n \"The assembly was amplified in Vial of One Shot® cells.\",\n \"The coding DNA sequence was then transferred into the PUHE25-2 expression vector (SphI cloning for the ATG and Bam HI for the stop).\",\n \"The base modification for Hb5 is therefore as follows: Nucleotide Sequence of Membrane-Bound Hb5 (SEQ ID No.\",\n \"3) 1 ATGGCAGAGC AGTCGGACGA GGCCGTGAAG TACTACACCC TAGAGGAGAT TCAGAAGCAC 61 AACCACAGCA AGAGCACCTG GCTGATCCTG CACCACAAGG TGTACGATTT GACCAAATTT 121 CTGGAAGAGC ATCCTGGTGG GGAAGAAGTT TTAAGGGAAC AAGCTGGAGG TGACGCTACT 181 GAGAACTTTG AGGATGTCGG GCACTCTACA GATGCCAGGG AAATGTCCAA AACATTCATC 241 ATTGGGGAGC TCCATCCAGA TGACAGACCA AAGTTAAACA AGCCTCCGGA AACTCTTATC 301 ACTACTATTG ATTCTAGTTC CAGTTGGTGG ACCAACTGGG TGATCCCTGC CATCTCTGCA 361 GTGGCCGTCG CCTTGATGTA TCGCCTATAC ATGGCAGAGG ACTGA Nucleotide Sequence of Soluble Hb5-Histidine Tag [Hb5-(His)4] (SEQ ID No.\",\n \"4) 1 ATGGCAGAGC AGTCGGACGA GGCCGTGAAG TACTACACCC TAGAGGAGAT TCAGAAGCAC 61 AACCACAGCA AGAGCACCTG GCTGATCCTG CACCACAAGG TGTACGATTT GACCAAATTT 121 CTGGAAGAGC ATCCTGGTGG GGAAGAAGTT TTAAGGGAAC AAGCTGGAGG TGACGCTACT 181 GAGAACTTTG AGGATGTCGG GCACTCTACA GATGCCAGGG AAATGTCCAG AACATTCATC 241 ATTGGGGAGC TCCATCCAGA TGACAGACCA AAGTTAAACA AGCCTCCGGA AACTCTTATC 301 ACTACTATTG ATTCTAGTTC CAGTAACGGA CATCACCACC ATTAA Amino Acid Sequence of [Hb5-(His)4] (SEQ ID No.\",\n \"5) 1 MLAEQSDEAV KYYTLEEIQK HNHSKSTWLI LHHKVYDLTK FLEEHPGGEE VLREQAGGDA 61 TENFEDVGHS TDAREMSRTF IIGELHPDDR PKLNKPPETL ITTIDSSSSN GHHHH 2.2) The Water-Soluble Protein-Histidine Tag with a Ser24→Cys mutation The chimeric protein obtained previously is then mutated in order to reveal a thiol group at the periphery of the protein, via a cysteine residue.\",\n \"This modification is produced by virtue of a site-directed mutagenesis on serine No.\",\n \"24 of Hb5(His)4.The mutagenesis kit used is the QuikChange® site-directed Mutagenesis Kit from Stratagene.\",\n \"The action of the pfu DNA polymerase makes it possible to reconstitute the expression vector and the sequence to be mutated.\",\n \"Supercompetent epicurian® XL1-blue bacteria are transformed with this plasmid.\",\n \"Several different locations for the mutation were tried, and position 24 is in particular exemplified, i.e.\",\n \"a modification of the serine residue to cysteine.\",\n \"The mutation was verified by sequencing in the laboratory (ABI sequencer): Nucleotide Sequence of Hb5-(His)4 mut24 (SEQ ID No.\",\n \"6) 1 ATGGCAGAGC AGTCGGACGA GGCCGTGAAG TACTACACCC TAGAGGAGAT TCAGAAGCAC 61 AACCACTGCA AGAGCACCTG GCTGATCCTG CACCACAAGG TGTACGATTT GACCAAATTT 121 CTGGAAGAGC ATCCTGGTGG GGAAGAAGTT TTAAGGGAAC AAGCTGGAGG TGACGCTACT 181 GAGAACTTTG AGGATGTCGG GCACTCTACA GATGCCAGGG AAATGTCCAG AACATTCATC 241 ATTGGGGAGC TCCATCCAGA TGACAGACCA AAGTTAAACA AGCCTCCGGA AACTCTTATC 301 ACTACTATTG ATTCTAGTTC CAGTAACGGA CATCACCACC ATTAA Amino Acid Sequence of Hb5-(His)4 mut24 (SEQ ID No.\",\n \"7) 1 MLAEQSDEAV KYYTLEEIQK HNHCKSTWLI LHHKVYDLTK FLEEHPGGEE VLREQAGGDA 61 TENFEDVGHS TDAREMSRTF IIGELHPDDR PKLNKPPETL ITTIDSSSSN GHHHH 2.3) Protein Expression in Escherichia coli The bacterial expression strain is XL1-blue.\",\n \"A preculture (6 h at 37° C. in 50 ml of medium) of a clone in a Luria-Bertami (LB) medium in the presence of a selection marker, ampicillin, made it possible to seed, in a proportion of 1/100, one liter of Terrific Broth medium supplemented with 100 mg/l of ampicillin.\",\n \"The culture is placed at ambient temperature with shaking for 48 h and then overexpression of protein is induced by adding isopropyl-β-D-thiogalactopyranoside (IPTG) in a final proportion of 0.5 mM.\",\n \"2.4) Protein Purification The bacteria are centrifuged after induction for 24-48 h and are frozen.\",\n \"A freezing/thawing cycle is followed by a lysis/fractionation step (lysozyme, cholic acid and ultrasonic disintegrator).\",\n \"The metal ion affinity chromatography (MIAC) purification procedure is defined as follows: An agarose gel functionalized with iminodiacetic acid (Sigma) is pre-loaded (flow of 1 ml/min) with nickel by loading a 50 mM solution of nickel chloride solubilized in an acetate buffer, pH 5.4.The column is washed with this same buffer at the same flow and the acetate is then replaced with a 50 mM sodium phosphate buffer, pH 7.The cell fractionation is directly injected onto the agarose gel column at 5 ml/min.\",\n \"Specifically, the chimeric proteins are retained on the column according to the MIAC technique.\",\n \"The column is subjected to a washing phase with phosphate buffer of increasing concentration of 50 to 250 mM, pH 7, until the solution passing through no longer exhibits an absorption spectrum in the ultraviolet range (240-300 nm).\",\n \"The protein sample is eluted during pulses of histines in solution at a concentration of 1 mg/ml in the 50 mM phosphate buffer, pH 7.Similar experiments were carried out on the yeast form of cytochrome b5.Thus, it is possible to obtain several mutant human and yeast chimeric proteins with a high degree of purity and in a proportion of several milligrams of protein.\",\n \"Example 3 Production of the Floating Protein Anchoring The hybrid bilayer structure presented in paragraph 1.3 was functionalized with the aim of conferring on the model metal ion affinity properties.\",\n \"To do this, a synthetic phospholipid having an imino-diacetate residue which complexes a nickel divalent cation was chosen.\",\n \"It is DOGS, or 1,2-dioleoyl-sn-glycero-3-[(N-(5-amino-1-carboxypentyl)iminodiacetic acid)succinyl] from Avanti polar Lipid, Inc.\",\n \"It has been demonstrated that such a molecule allows the interaction of proteins having a polyhistidine domain in their primary sequence.\",\n \"With the aim of introducing these synthetic phospholipids into the membrane model, we produced lipid vesicles composed of a mixture of lipids containing 10% (mol/mol) of modified lipid.\",\n \"These mixed liposomes have the same behavior with respect to the hydrophobic support as those composed of a single species of phospholipids.\",\n \"The fusion results by SPR give rise to values which are comparable in terms of kinetics of fusion and of covering (results not shown).\",\n \"A method for addressing soluble proteins to membrane supports in a stable and specific manner is to use the coupling between a chelating divalent metal cation present at the surface of the membrane and histidine residues.\",\n \"It has been demonstrated that cytochrome b5 in which the membrane domain has been deleted in favor of a histidine Tag segment conserves an affinity for biological membranes displaying chelating divalent cations.\",\n \"Various forms of cytochrome b5 with a HisTag domain were produced (paragraph 2.1).\",\n \"The immobilization of the cytochromes b5 on the hybrid bilayer via an MIAC approach was characterized by SPR (FIG.\",\n \"2).\",\n \"To evaluate the binding capacity of the membrane, multiple protein injections were performed within its surroundings.\",\n \"The protein concentrations are conventionally between 0.5 and 2 μM and produce biosensor saturation kinetics.\",\n \"For the Hb5(His)4 form, a maximum load signal of 650 RU, which corresponds to approximately 75% of optimum covering on such a membrane model (FIG.\",\n \"1a), was obtained.\",\n \"To demonstrate the specific interaction of these proteins with the biosensor, multiple injections of an agent which competes with histidine-Ni2+ coordinations were performed.\",\n \"A solution of histidine at 1 mg/ml injected at the surface of the biosensor produces a large decrease in the signal, i.e.\",\n \"in the amount of cytochrome b5 attached to the membrane (FIG.\",\n \"2b).\",\n \"In this way, it is possible to compete with the specific interaction and to return to the signal base line.\",\n \"This procedure, using an aqueous solution which is nondenaturing for the biomolecules, thus enables complete regeneration of the surface of the lipid monolayer.\",\n \"The results given in FIG.\",\n \"2 confirm the strong potential of the biosensor in terms of protein capture with a high degree of compactness.\",\n \"They make it possible to underline the very high level of control of the authors over the various phases of construction of this supramolecular assembly since, by modulating the protein or competitor injection pulses, it is possible to precisely attach the amount of protein at the surface.\",\n \"This great modularity can be summarized in FIG.\",\n \"3.Example 4 Protein-ssDNA Grafting Via a Cis-Pt Complex Platinum complexes, in particular cis-diamine-dichloroplatinums (cis-[PtCl2(NH3)2]) are agents which bind covalently to DNA.\",\n \"In addition, these molecules, by substitution of their labile chloro ligand, can couple, firstly, a base of a DNA and, secondly, a nucleophilic residue of an amino acid of a protein.\",\n \"4.1) Materials Cis- [PtCl2(NH3)2] and trans- [ PtCl2 (NH3)2] are provided by Johnson Matthey (London, United Kingdom).\",\n \"Cis-[Pt(NH3)2 (H2O)2] 2+ and trans- [Pt(NH3)2 (H2O)2] 2+are prepared by dissolving a suspension of cis-[Pt(NO3)2 (NH3)2] and trans- [Pt(NO3)2 (NH3)2] in water, respectively formed by reacting cis-[PtCl2(NH3)2] and trans-[PtCl2(NH3)2] with silver nitrate.\",\n \"The 20-mer DNA, its complementary strand and its biotinylated complementary strand come from Eurogentec.\",\n \"The sequence of the 20-mer DNA, 5′CTATCATTTGCTTACTATTC 3′(SEQ ID No.\",\n \"13), is chosen such that it possesses only one guanine since the N7 nitrogen is the main site of platination of DNA (Lepre et al., 1990).\",\n \"4.2) Formation of the DNA-b5 Bridging Via Platinum Bridging The platination reactions are carried out firstly on the DNA or on the cytochrome b5 (Hb5(His)4 and Hb5(His)4mut24), but the DNA-b5 bridging yield is greater when the cytochrome b5 is platinated first.\",\n \"Platination of the ssDNA (500 μM) with one equivalent of platinum complex in 70 mM NaClO4 leads to the formation of the cis- or trans- [Pt(NH3)2(H2O) ]2+ monoadduct linked to the guanine.\",\n \"The platination site is determined after treatment of the modified oligonucleotide with piperidine-DMS under “Maxam-Gilbert” sequencing conditions followed by treatment with NaCN.\",\n \"A lack of reactivity of the platinated N7 guanine with the dimethyl sulfate (DMS) makes it possible to conclude that the latter is protected with respect to the DMS and is therefore effectively platinated (Comess et al., 1990).\",\n \"The cytochrome b5 (100-200 μM) is incubated in the presence of 5 equivalents of platinum complexes in unbuffered water (pH 5) for 5 hours at 25° C. The cytochrome b5 is then incubated (90 μM) with 1.5 equivalents of DNA in unbuffered water (pH 5) for 50 hours at 25° C. The DNA-b5 complexes are separated from the non-complexed DNA molecules using an agarose gel functionalized with Ni2+/iminodiacetic acid groups (Sigma).\",\n \"The protein is eluted by injection of imidazole (1 M).\",\n \"The excess DNA is not retained on the column.\",\n \"The DNA-b5 complexes coupled via a platinum bridge are also purified from the nonreactive forms of DNA using streptavidin-coated magnetic spheres from Promega, according to a modification of the procedure described by Promega: 0.5 nmol of total DNA (complexed or non-complexed) of the complexation mixture is heated at 45° C. for 10 min in water and is then brought into contact with its biotinylated complementary strand in 0.075 M NaCl, 0.0075 M citrate, pH 7.2, at 2.5° C. The mixture (500 μl) is added to 0.6 mg of magnetic spheres complexed with streptavidin, pre-washed and equilibrated in the same buffer at 2.5° C. The ssDNA-b5 platinum complexes are not retained on the spheres and the non-complexed ssDNA is eluted between 30-40° C. in 0.015 M NaCl and 0.0015 M citrate, pH 7.2.These purification steps are followed, firstly, by detection of the radioactivity of the ssDNA labeled with 32P in the 5′ position (radiolabeled beforehand via the action of polynucleotide kinase using [γ32P]ATP) and, secondly, by a spectral study over a window of absorbance comprising 413 nm for the b5 (e=100 000 M−1.cm−1) and 260 nm for the ssDNA (e=180500 M−1.cm−1) .\",\n \"The latter is corrected for the Hb5 absorbance component at this wavelength (i.e.\",\n \"⅕ of its absorbance at 413 nm).\",\n \"However, it is specified that these last two procedures do not make it possible to separate the “ssDNA-b5” platinum complex from the non-complexed cytochrome b5.The cis- [Pt(NH3)2(H2O)2]2+ and trans-[Pt(NH3)2(H2O)2]2+ platinum complexes are capable of coupling the cytochrome b5 with the 20-mer ssDNA.\",\n \"The platination site on the DNA oligonucleotide occurs at N7 of a unique guanine.\",\n \"The “DNA-b5” macromolecular complexes are characterized by gel electrophoresis analyses under denaturing conditions, spectro-photometric quantification analyses and determination of melting temperature with a complementary free ssDNA.\",\n \"An electrophoretic study on SDS-15% PAGE of the mixture composed of the 5′-radiolabeled ssDNA, of Hb5[His]4 and of cis- or trans- [Pt(NH3)2(H2O)2] 2+was carried out.\",\n \"After migration of the samples, the radioactivity of the ssDNA is associated with the Hb5[His]4 migration band (revealed by coomassie blue staining), demonstrating that a covalent bond has been formed between the 20-mer ssDNA and the cytochrome b5 (FIG.\",\n \"4).\",\n \"The “DNA-pt-b5” complex was characterized by spectral analysis with the aim of determining the stoichiometry of the reaction between the cytochrome b5 and the 20-mer ssDNA.\",\n \"The experimental conditions allow the formation of an equimolecular complex as indicated by the respective absorbance of the cytochrome b5 at 413 nm and of the 20-mer ssDNA at 260 nm (FIG.\",\n \"5).\",\n \"4.3) Determination of the Melting Temperature The melting temperatures of the 20-mer ssDNA and of the 20-mer ssDNA coupled to the b5 via a cis-[Pt(NH3)2 (H2O) 2] 2+bridge were determined in 0.1 M NaClO4, pH 4, with their complementary ssDNAs.\",\n \"The melting temperatures were measured on a Uvikon 941 spectrophotometer (Kontron).\",\n \"0.2 nmol of 20-mer ssDNA (or “ssDNA-b5-Pt” complex) were incubated with an ssDNA having a complementary sequence, at a final concentration of 0.5 μM, in 0.025 M NaClO4, pH 4.4.The duplex sample is heated to 80° C. (20-mer DNA) or 65° C. (“DNA-Pt-b5”), the temperature is subjected to a decrease of 1° C./min down to 2.5° C., and the absorbance is measured at 260 nm.\",\n \"The samples are heated from 2.5° C. to 60° C. at 1° C./min and their absorbance is followed at 260 nm, with the aim of verifying that the curves obtained during the cooling and the heating can be superimposed (reversible profiles).\",\n \"The Tm values were determined using Kontron software.\",\n \"The melting temperature of the duplex containing the 20-mer ssDNA decreases by 22° C. (from 45° C. to 23° C.) when it is coupled with the cytochrome b5 (FIG.\",\n \"6).\",\n \"No difference was observed between the complex comprising Hb5(His)4 and the cysteine mutant; the cysteine at position 24 does not appear, for the cis-Pt complex, to be more reactive than the other nucleophilic sites present at the periphery of the cytochrome b5.Be that as it may, the platination reactions do not occur on the 4-histidine tag in the carboxy-terminal position, in which case the complex thus formed could not be purified on an agarose-imino-diacetate column or would not be capable of interacting strongly with the cell.\",\n \"Experiments for characterizing the various complexes by mass spectroscopy (electrospray, Maldi-TOF) are in the process of being carried out.\",\n \"Example 5 Protein-ssDNA Grafting Via a Spacer Arm 5.1) Functionalization of ssDNA with DPDPB A second alternative was explored, concerning the grafting of a nucleic acid with the cytochrome b5.With a view to optimum display of the DNA coupled at the surface of the membrane biosensor, the inventors chose to use not direct coupling, but preferentially complexation via a spacer arm.\",\n \"The molecule providing the intermolecular bridging is 1,4-di-[3′-(2′-pyridyl-dithio)propionamido]butane, or DPDPB.\",\n \"This molecule allows bonding of the covalent type between two molecules having thiol functions and provides an intermolecular space of 1.6 nm.\",\n \"In order to produce such a supramolecular assembly, a thiol function was created both on an oligonucleotide and on the cytochrome b5.The modified oligonucleotides are of HPLC quality and are produced by Eurogentec.\",\n \"The various modified oligonucleotides are as follows: Oligonucleotide No.\",\n \"W1 (SEQ ID No.\",\n \"8) 5′GCT-AGC-TGC-ATA-GAT-CTC-TAC-C-SH 3′ The thiol modification is performed at the 3′ end of the 22-mer.\",\n \"Oligonucleotide No.\",\n \"W2 (SEQ ID No.\",\n \"9) 5′ GCT-AGC-TGC-ATA-GAT-CTC-TAC-C 3′ The thiol modification is performed intra-chain on a thymine placed at position 11 of the 22-mer.\",\n \"The various nonmodified oligonucleotides involved in the exemplification are as follows: Oligonucleotide No.\",\n \"W3 (SEQ ID No.\",\n \"10) 5′ GGT-AGA-GAT-CTA-TGC-AGC-TAG-G-3′ W3 is the 22-mer complementary to W1.Oligonucleotide No.\",\n \"W4 (SEQ ID No.\",\n \"11) 5′-GCA-GCT-AGC-GGT-AGA-GAT-CT-3′ W4 has a sequence complementary to the first 9 bases of Wl (5′ side) and to the first 11 bases of W1 (3′ side).\",\n \"In this manner, W4 has the particularity of being able to bridge two W1 oligonucleotides.\",\n \"Oligonucleotide No.\",\n \"W5 (SEQ ID No.\",\n \"12) 5′-GCT-AGC-TGC-ATA-GAT-CTC-TAC-C-3′ W5 corresponds to the random base sequence of W1.The DPDPB-W1 coupling procedure is as follows: in order to prevent the dimerization of the W1s by disulfide bonding, the oligonucleotides are treated with a dithiothreitol (DTT) reducing agent, in a molar fraction proportion of ⅕.\",\n \"The mixture is incubated at 30° C. for 15 min.\",\n \"The stock solution of DPDPB is solubilized in dimethyl sulfoxide (DMSO) in a proportion of 10 mg/ml.\",\n \"DPDPB, in a proportion of 1 mol of W1 per 500 to 5 000 mol of DPDPB, is added in the presence of the residual DTT, for an incubation time of 2 to 4 hours at 30° C. The W1 sample is then specifically attached to a DEAE ion exchange column, the reagents are eliminated, and the oligonucleotide is eluted by adding a 1.5 M NaCl solution.\",\n \"The degree of modification can be calculated by assaying the cross-linking agent with DTT.\",\n \"This reduction causes the second thiopyridine group of the bridging agent to leave, and it can be quantified spectrally at 341 nm.\",\n \"The procedure described makes it possible to achieve 100% of complex.\",\n \"5.2) Cytochrome b5-W1 Grafting Via the Spacer Arm In order to perform controlled and reversible grafting on the cytochrome b5, the inventors carried out a site-directed mutagenesis of Hb5(His)4 as described in paragraph 2.2, producing the protein Hb5(His)4mutant24.The serine→cysteine mutation makes it possible to specifically introduce a thiol group at the periphery of the protein.\",\n \"In order to prevent dimerization of the proteins, the solution is treated with DTT (Xmol b5/DTT=⅕) for 30 min at 30° C. The sample is then loaded onto a gel exclusion chromatography column (G25) so as to remove the DTT and is immediately injected into the solution of modified oligonucleotides (Xmol b5/W1=½) for 4 h at 30° C. The excess oligonucleotide is removed by retention of the proteins on an iminodiacetate-Ni2+gel.\",\n \"Elution of the proteins is obtained by pulses of a 1 mg/ml histidine solution, as indicated during the HisTag protein purification step.\",\n \"Spectral analysis over a [240-500] nm window makes it possible to demonstrate the effective coupling of Hb5(His)4mut24 with W1 (FIG.\",\n \"7).\",\n \"With regard to the molar absorption coefficients of the two macromolecules, the degrees of complexation range from 35 to 65%.\",\n \"In order to subsequently use only an equimolecular Hb5/W1 population, an additional purification step is carried out on DEAE.\",\n \"Elution performed under an NaCl concentration gradient makes it possible to isolate the various species reconstituted during the coupling (monomeric and dimeric Hb5, Hb5-spacer-W1).\",\n \"Thus, all the species are separated from the initial population, by an increasing NaCl gradient (FIG.\",\n \"7): Spectrum a) Hb5(His)4mut24 monomer Spectrum b) Hb5(His)4mut24 dimer Spectrum c) equimolar b5-W1 complex.\",\n \"According to this procedure, the b5-W1 population is isolated and can be brought into contact with the hybrid membrane.\",\n \"Example 6 Preparation of the Supramolecular Assembly “b5-cis Pt-ssDNA” and “b5-DPDPB-ssDNA” The grafting of the various complexes onto the hybrid bilayer was characterized by surface plasmon resonance in a BIAcore device.\",\n \"Radioactivity-based and fluorescence labeling-based experiments are in the process of being developed in the laboratory.\",\n \"The experiments consisting of coupling the molecular assembly to the lipid model by virtue of the chelating nickel complex by surface plasmon resonance made it possible to show the formation of a stable complex.\",\n \"The levels of compactness obtained with the assembly appear, on the other hand, to be less than those obtained with the protein not modified with the oligonucleotide.\",\n \"The increase in negative charges introduced by W1 may be responsible for repulsions between supramolecular blocks and may thus lead to less covering.\",\n \"The covering values were evaluated at between 200 and 250 RU.\",\n \"It was verified that, like Hb5(His)4, the structures are interacting specifically with the support and that the action of agents which compete with the chelation, such as a 1 mg/ml histidine solution, allows complete regeneration of the structure (data not shown, but exhibiting the same behavior as those appearing in FIG.\",\n \"2).\",\n \"Example 7 Potential of the Biosensor and Detection of Hybridization In order to evaluate the potential of the membrane biosensor in terms of specificity and sensitivity, various tests were carried out.\",\n \"Firstly, it was essential to verify the absence of nonspecific interactions of oligonucleotides with the biosensor.\",\n \"To do this, various injections of W4 were performed.\",\n \"W4 represents a random sequence from the bases constituting W1; studying the evolution of the signal made it possible to show that W4 does not interact at the surface of the biosensor, i.e.\",\n \"the lipid structure and the b5-W1 supramolecular assembly, whatever the bridging means (cis-Pt or DPDPB).\",\n \"Secondly, various injections of W3 were performed at various salt concentrations.\",\n \"Thus, it was clearly demonstrated that the injections of W3, the sequence complementary to W1, in the presence of a high concentration of NaCl were accompanied by association or hybridization kinetics.\",\n \"On the other hand, for concentrations of NaCl less than or equal to 20 mM, the same oligonucleotide interacts very little or not at all with the biosensor.\",\n \"These results demonstrate the possibility of detecting hybridization states between two ssDNAs at the surface of the biosensor.\",\n \"The hybridization results show a threshold value of approximately 30 to 35 RU.\",\n \"Now, according to the various methods of calibration, the value expected for complete hybridization based on a b5-W1 block signal of 250 RU is 70 RU.\",\n \"However, the Tm measurements made on W1 clearly show a phase transition temperature close to ambient temperature, i.e.\",\n \"the temperature at which measurements are made in the BIAcore device.\",\n \"At this temperature, only 50% of the hybridization is expected.\",\n \"The value measured with our device is therefore close to optimum hybridization, which is proof of a very favorable accessibility of the target molecules displayed by the biosensor.\",\n \"The size of the working cell within the BIAcore device is of the order of 1 mm2.However, the surface really probed by the damped wave represents only 0.224 mm2 Considering the calibration 1 000 RU←→0.224 ng/0.224 mm2, 250 RU of molecular assembly correspond to 0.1 ng, i.e.\",\n \"5 fmol ≈3×109 molecules.\",\n \"The membrane biosensor allows the molecular hybridization of approximately 109 molecules.\",\n \"In addition, the biosensor presented requires only the capture of a complementary ssDNA, without it being necessary to use a labeled probe.\",\n \"Bibliographical References Berney et al.\",\n \"(2000) Sensors amd Actuators B 68, 100-108 Boncheva et al.\",\n \"(1999) Langmuir 15, 4317-4320 Boon et al.\",\n \"(2000) Natural Biotechnology 18, 1096-1100 Braun et al.\",\n \"(1998) Nature 391, 775-778 Cheng et al (2000) Reviews in molecular biotechnology 74, 159-174 (p 18) Comess et al.\",\n \"(1990) Biochemistry, 29, 2102-2110 Ebbesen et al.\",\n \"(1998) Nature 391, 667-669 (p 18) Gulik-Krzywicki et al.\",\n \"(1967) J Mol Biol; 27(2): 303-22 Heyse et al.\",\n \"(1998) Biochemistry 37, 507-522 Kalb et al.\",\n \"(1992) Biochim.\",\n \"Biophys.\",\n \"Acta 1103: 307-316 Lepre et al.\",\n \"(1990 ) Nucleic Acids Molec.\",\n \"Biol., 4, 9-38 Marchal et al.\",\n \"(1998) Biophysical Journal 74, 1937-1948 Steel et al.\",\n \"(2000) Biophysical Journal 79, 975-981 Wittung-Stafshede et al.\",\n \"(2000) Colloids and Surfaces 174, 269-273\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10474299"}}},{"rowIdx":796,"cells":{"text":{"kind":"list like","value":[["Server-based browser system","A server-based browser system provides a server-based browser and a client system browser.","The client browser reports the position of a user click on its display screen, hotspot ID, or URL to the server-side browser which retrieves a Document Object Model (DOM) model and view tree for the client and finds the location on the Web page that the user clicked on using the coordinates or hotspot ID received from the client.","If there is a script associated with the location, it is executed and the resulting page location is requested from the appropriate server.","If there is a URL associated with the location, it is requested from the appropriate server.","The response Web page HTML definition is parsed and a DOM tree model is created which is used to create a view tree model.","The server-side browser retrieves a style sheet, layout algorithms, and device constraints for the client device and lays out the view model using them onto a virtual page and determines the visual content.","Textual and positional information are highly compressed and formatted into a stream and sent to the client browser which decodes the stream and displays the page to the user using the textual and positional information."],["1.A process for providing a server-based Web browser that interacts with a plurality of client device types, comprising the steps of: (a) providing browsing means on a server for accepting a Web page from a Web server, said browsing means comprising Web page layout means for laying out said Web page in a two-dimensional virtual page, said Web page layout means determining what information is visible on said virtual page, said Web page layout means calculating positioning of said visible information within said Web page, and said browsing means creating a compressed stream containing said visible information and the positioning information of said visible information; (b) providing client browsing means on a client system for accepting compressed streams representing display pages; and (c) providing means on said server for sending said compressed stream to said client browsing means; wherein said client browsing means decompresses said compressed stream and displays said visible information using said positioning information.","2.The process of claim 1, wherein said browsing means comprises: a database of style sheets on a storage device for supported client devices; a database of layout algorithms on said storage device for supported client devices; and a database of device constraint information on said storage device for supported client devices; wherein said Web page layout means retrieves a style sheet, layout algorithms, and device constraints from said storage device for said client system; and wherein said Web page layout means lays out said Web page based on said client system's style sheet, layout algorithms, and device constraints.","3.The process of claim 1, wherein said step of providing browsing means further comprises the sub-steps of: parsing said Web page's HTML code to create a Document Object Model (DOM) tree; creating a view tree using said DOM tree; and storing said DOM tree and said view tree on a storage device for said client system.","4.The process of claim 3, wherein said client browsing means accepts a URL from a user or determines a position or hotspot ID of a user's click or press on a display page, wherein said client browsing means sends said URL, said position, or said hotspot ID to said browsing means, and wherein said browsing means accepts said URL, said position, or said hotspot ID from said client browsing means.","5.The process of claim 4, wherein said step of providing browsing means further comprises the sub-steps of: retrieving a DOM tree and view tree from said storage device for said client system; determining the location of said position or hotspot ID on said Web page using said view tree; finding a script or URL associated with the location using said DOM tree; wherein if a script is associated with the location then executing the script and determining the resulting URL; and requesting the Web page associated with said resulting URL or said associated URL from the appropriate server.","6.The process of claim 3, wherein said step of providing browsing means further comprises the sub-step of: saving said client system's state information on said storage device for a particular user; wherein said client system's state information includes Javascripts and cookies; and wherein said client system's state information allows said browsing means to track the current state of said client system and the particular user.","7.A process for providing a cross platform Web browser that interacts with a plurality of client device types, comprising the steps of: (a) providing browsing means on a server for accepting a Web page from a Web server; said step pf providing browsing means further comprising the sub-steps of: parsing said Web page's HTML code to create a Document Object Model (DOM) tree; creating a view tree using said DOM tree; providing Web page layout means for laying out said Web page in a two-dimensional virtual page; wherein said Web page layout means determines what information is visible on said virtual page; wherein said Web page layout means removes unnecessary structural information and retains visible information in said view tree; and storing said DOM tree and said view tree on a storage device for said client system; wherein said browsing means creates a compressed stream containing said DOM tree and said view tree; (b) providing client browsing means on a client system for accepting compressed streams; and (c) providing means on said server for sending said compressed stream to said client browsing means; wherein said client browsing means decompresses said compressed stream and displays said visible information using said DOM tree and said view tree.","8.The process of claim 7, wherein said browsing means comprises: a database of style sheets on said storage device for supported client devices; a database of layout algorithms on said storage device for supported client devices; and a database of device constraint information on said storage device for supported client devices; wherein said Web page layout means retrieves a style sheet, layout algorithms, and device constraints from said storage device for said client system; and wherein said Web page layout means lays out said Web page based on said client system's style sheet, layout algorithms, and device constraints.","9.The process of claim 7, wherein said client browsing means accepts a URL from a user or determines a position or hotspot ID of a user's click or press on a display page, wherein said client browsing means sends said URL, said position, or said hotspot ID to said browsing means, and wherein said browsing means accepts said URL, said position, or said hotspot ID from said client browsing means.","10.The process of claim 9, wherein said step of providing browsing means further comprises the sub-steps of: retrieving a DOM tree and view tree from said storage device for said client system; determining the location of said position or hotspot ID on said Web page using said view tree; and finding a script or URL associated with the location using said DOM tree; wherein if a script is associated with the location then executing the script and determining the resulting URL; and requesting the Web page associated with said resulting URL or said associated URL from the appropriate server.","11.The process of claim 7, wherein said step of providing browsing means further comprising the sub-step of: saving said client system's state information on said storage device for a particular user; wherein said client system's state information includes Javascripts and cookies; and wherein said client system's state information allows said browsing means to track the current state of said client system and the particular user.","12.An apparatus for a server-based Web browser that interacts with a plurality of client device types, comprising: (a) browsing means on a server for accepting a Web page from a Web server, said browsing means further comprising Web page layout means for laying out said Web page in a two-dimensional virtual page, said Web page layout means determining what information is visible on said virtual page, and said Web page layout means calculating positioning of said visible information within said Web page; wherein said browsing means creates a compressed stream containing said visible information and the positioning information of said visible information; (b) client browsing means on a client system for accepting compressed streams representing display pages; and (c) means on said server for sending said compressed stream to said client browsing means; wherein said client browsing means decompresses said compressed stream and displays said visible information using said positioning information.","13.The apparatus of claim 12, wherein said browsing means further comprises: a database of style sheets on a storage device for supported client devices; a database of layout algorithms on said storage device for supported client devices; and a database of device constraint information on said storage device for supported client devices; wherein said Web page layout means retrieves a style sheet, layout algorithms, and device constraints from said storage device for said client system; and wherein said Web page layout means lays out said Web page based on said client system's style sheet, layout algorithms, and device constraints.","14.The apparatus of claim 12, wherein said browsing means further comprises: a module for parsing said Web page's HTML code to create a Document Object Model (DOM) tree; a module for creating a view tree using said DOM tree; and a module for storing said DOM tree and said view tree on a storage device for said client system.","15.The apparatus of claim 14, wherein said client browsing means accepts a URL from a user or determines a position or hotspot ID of a user's click or press on a display page, wherein said client browsing means sends said URL, said position, or said hotspot ID to said browsing means, and wherein said browsing means accepts said URL, said position, or said hotspot ID from said client browsing means.","16.The apparatus of claim 15, wherein said browsing means further comprises: a module for retrieving a DOM tree and view tree from said storage device for said client system; a module for determining the location of said position or hotspot ID on said Web page using said view tree; a module for finding a script or URL associated with the location using said DOM tree; wherein if a script is associated with the location then executing the script and determining the resulting URL; and a module for requesting the Web page associated with said resulting URL or said associated URL from the appropriate server.","17.The apparatus of claim 14, wherein said browsing means further comprises: a module for saving said client system's state information on said storage device for a particular user; wherein said client system's state information includes Javascripts and cookies; and wherein said client system's state information allows said browsing means to track the current state of said client system and the particular user.","18.An apparatus for a cross platform Web browser that interacts with a plurality of client device types, comprising: (a) browsing means on a server for accepting a Web page from a Web server; said browsing means further comprising: a module for parsing said Web page's HTML code to create a Document Object Model (DOM) tree; a module for creating a view tree using said DOM tree; Web page layout means for laying out said Web page in a two-dimensional virtual page; wherein said Web page layout means determines what information is visible on said virtual page; wherein said Web page layout means removes unnecessary structural information and retains visible information in said view tree; and a module for storing said DOM tree and said view tree on a storage device for said client system; wherein said browsing means creates a compressed stream containing said DOM tree and said view tree; (b) client browsing means on a client system for accepting compressed streams; (c) means on said server for sending said compressed stream to said client browsing means; and wherein said client browsing means decompresses said compressed stream and displays said visible information using said DOM tree and said view tree.","19.The apparatus of claim 18, wherein said browsing means further comprises: a database of style sheets on said storage device for supported client devices; a database of layout algorithms on said storage device for supported client devices; a database of device constraint information on said storage device for supported client devices; wherein said Web page layout means retrieves a style sheet, layout algorithms, and device constraints from said storage device for said client system; and wherein said Web page layout means lays out said Web page based on said client system's style sheet, layout algorithms, and device constraints.","20.The apparatus of claim 18, wherein said client browsing means accepts a URL from a user or determines a position or hotspot ID of a user's click or press on a display page, wherein said client browsing means sends said URL, said position, or said hotspot ID to said browsing means, and wherein said browsing means accepts said URL, said position, or said hotspot ID from said client browsing means.","21.The apparatus of claim 20, wherein said browsing means further comprises: a module for retrieving a DOM tree and view tree from said storage device for said client system; a module for determining the location of said position or hotspot ID on said Web page using said view tree; a module for finding a script or URL associated with the location using said DOM tree; wherein if a script is associated with the location then executing the script and determining the resulting URL; and a module for requesting the Web page associated with said resulting URL or said associated URL from the appropriate server.","22.The apparatus of claim 18, wherein said browsing means further comprises: a module for saving said client system's state information on said storage device for a particular user; wherein said client system's state information includes Javascripts and cookies; and wherein said client system's state information allows said browsing means to track the current state of said client system and the particular user.","23.A program storage medium readable by a computer, tangibly embodying a program of instructions executable by the computer to perform method steps for a server-based Web browser that interacts with a plurality of client device types, comprising the steps of: (a) providing browsing means on a server for accepting a Web page from a Web server, said browsing means providing Web page layout means for laying out said Web page in a two-dimensional virtual page, said Web page layout means determining what information is visible on said virtual page, said Web page layout means calculating positioning of said visible information within said Web page; and said browsing means creating a compressed stream containing said visible information and the positioning information of said visible information; (b) providing client browsing means on a client system for accepting compressed streams representing display pages; and (c) providing means on said server for sending said compressed stream to said client browsing means; wherein said client browsing means decompresses said compressed stream and displays said visible information using said positioning information.","24.The program storage medium of claim 23, wherein said browsing means comprises: a database of style sheets on a storage device for supported client devices; a database of layout algorithms on said storage device for supported client devices; and a database of device constraint information on said storage device for supported client devices; wherein said Web page layout means retrieves a style sheet, layout algorithms, and device constraints from said storage device for said client system; and wherein said Web page layout means lays out said Web page based on said client system's style sheet, layout algorithms, and device constraints.","25.The program storage medium of claim 23, wherein said step of providing browsing means further comprising the sub-steps of: parsing said Web page's HTML code to create a Document Object Model (DOM) tree; creating a view tree using said DOM tree; and storing said DOM tree and said view tree on a storage device for said client system.","26.The program storage medium of claim 25, wherein said client browsing means accepts a URL from a user or determines a position or hotspot ID of a user's click or press on a display page, wherein said client browsing means sends said URL, said position, or said hotspot ID to said browsing means, and wherein said browsing means accepts said URL, said position, or said hotspot ID from said client browsing means.","27.The program storage medium of claim 26, wherein said step of providing browsing means further comprising the sub-steps of: retrieving a DOM tree and view tree from said storage device for said client system; determining the location of said position or hotspot ID on said Web page using said view tree; finding a script or URL associated with the location using said DOM tree; wherein if a script is associated with the location then executing the script and determining the resulting URL; and requesting the Web page associated with said resulting URL or said associated URL from the appropriate server.","28.The program storage medium of claim 23, wherein said step of providing browsing means further comprising the step of: saving said client system's state information on said storage device for a particular user; wherein said client system's state information includes Javascripts and cookies; and wherein said client system's state information allows said browsing means to track the current state of said client system and the particular user.","29.A program storage medium readable by a computer, tangibly embodying a program of instructions executable by the computer to perform method steps for a cross platform Web browser that interacts with a plurality of client device types, comprising the steps of: (a) providing browsing means on a server for accepting a Web page from a Web server; said step of providing browsing means further comprising the sub-steps of: parsing said Web page's HTML code to create a Document Object Model (DOM) tree; creating a view tree using said DOM tree; providing Web page layout means for laying out said Web page in a two-dimensional virtual page; wherein said Web page layout means determines what information is visible on said virtual page; wherein said Web page layout means removes unnecessary structural information and retains visible information in said view tree; and storing said DOM tree and said view tree on a storage device for said client system; wherein said browsing means creates a compressed stream containing said DOM tree and said view tree; (b) providing client browsing means on a client system for accepting compressed streams; and (c) providing means on said server for sending said compressed stream to said client browsing means; wherein said client browsing means decompresses said compressed stream and displays said visible information using said DOM tree and said view tree.","30.The program storage medium of claim 29, wherein said browsing means comprises: a database of style sheets on said storage device for supported client devices; a database of layout algorithms on said storage device for supported client devices; and a database of device constraint information on said storage device for supported client devices; wherein said Web page layout means retrieves a style sheet, layout algorithms, and device constraints from said storage device for said client system; and wherein said Web page layout means lays out said Web page based on said client system's style sheet, layout algorithms, and device constraints.","31.The program storage medium of claim 29, wherein said client browsing means accepts a URL from a user or determines a position or hotspot ID of a user's click or press on a display page, wherein said client browsing means sends said URL, said position, or said hotspot ID to said browsing means, and wherein said browsing means accepts said URL, said position, or said hotspot ID from said client browsing means.","32.The program storage medium of claim 31, wherein said step of providing browsing means further comprising the steps of: retrieving a DOM tree and view tree from said storage device for said client system; determining the location of said position or hotspot ID on said Web page using said view tree; finding a script or URL associated with the location using said DOM tree; wherein if a script is associated with the location then executing the script and determining the resulting URL; and requesting the Web page associated with said resulting URL or said associated URL from the appropriate server.","33.The program storage medium of claim 29, wherein said step of providing browsing means further comprises the sub-step of: saving said client system's state information on said storage device for a particular user; wherein said client system's state information includes Javascripts and cookies; and wherein said client system's state information allows said browsing means to track the current state of said client system and the particular user."],["<SOH> BACKGROUND OF THE INVENTION <EOH>1.Technical Field The invention relates to Web browsing technology.","More particularly, the invention relates to a system and method for fast delivery of and fast rendering of Web pages on a lightweight network device.","2.Description of the Prior Art A typical Web page today contains not only HTML content but also, by reference to, other content elements such as graphics and external style sheets.","A browser acquires the referenced content elements by spawning worker threads that retrieve the data via multiple HTTP-TCP/IP connections.","This model works well for desktop systems containing large amounts of processing power and RAM.","However, it is usually not feasible for mobile, lightweight, embedded devices in which processing power and RAM are much limited.","The process for proper HTML parsing, validation, and layout code is normally quite complex and requires large amount of CPU and RAM to execute.","Size and manufacturing cost have been major concerns in providing embedded devices with a capacity to implement the process.","Although embedded devices are getting more and more processing power and RAM, the requirements of a proper browser engine still outdistance the resources available to even the highest end of embedded devices.","High levels of CPU usage are detrimental to the preservation of battery life, which is a very important consideration for today's mobile devices.","Heavy loads in HTTP communications, HTML parsing, HTML validation and layout may largely shorten the battery life.","Bandwidth usage is also a major constraint that must be considered in designing a browser model for embedded devices.","Text based HTML content, although compressible, is already quite large with respect to the limited transmission speed of today's wireless networks.","When image content designed for higher resolution and larger form factor display is included, one may quickly end up consuming a lot of wasted bandwidth transmitting redundant, not necessarily usable data to mobile devices.","Although various browsing technologies have been already presented in the marketplace, many of them fail to meet the tight constraints and stringent requirements of the embedded devices.","Writing a cross platform code base for these devices is an even bigger challenge, given the multitude of operating systems, memory models and processors available for embedded device manufacturers.","It would be advantageous to provide a server-based browser system that offloads heavy weight components of a traditional browser to a back end server.","It would further be advantageous to provide a server-based browser system that leaves a lightweight client device with a minimal task of implementing a customizable interface to display a pre-rendered representation of Web pages."],["<SOH> SUMMARY OF THE INVENTION <EOH>The invention provides a server-based browser system.","The system offloads heavy weight components of a traditional browser to a back end server.","In addition, the invention leaves a lightweight client device with a minimal task of implementing a customizable interface to display a pre-rendered representation of Web pages.","A preferred embodiment of the invention provides a server-based browser and a client system browser.","The client browser determines the position of a user click on its display screen and sends a navigation request containing the coordinates of the dick or a hotspot ID, to the server-side browser.","The client browser will also accept a specific URL from a user and places the URL in the navigation request.","The server-side browser retrieves a Document Object Model (DOM) model and view tree from a storage device for the client It then finds the location on the Web page that the user clicked on using the coordinates or hotspot ID received from the client.","If there is a script associated with the location, it is executed and the resulting page location is requested from the appropriate server.","If there is a URL associated with the location, it is requested from the appropriate server.","The response Web page HTML page definition is parsed and a DOM tree model is created.","The server-side browser uses the DOM tree and creates a two-dimensional view tree model, and stores both trees on the storage device.","The server-side browser retrieves a style sheet, layout algorithms, and device constraints for the client device from the storage device.","It lays out the view model using the style sheet, layout algorithms, and device constraints onto a virtual page.","If the client browser is a subset of the server-side browser engine, then the DOM tree and view tree are compressed into a stream and sent to the client browser which decodes the stream, translates the DOM tree and view tree, and displays the page to the user.","Otherwise, the server-side browser determines the visual content on the virtual page and the location of the visual content.","Visual content and positional information are highly compressed and formatted into a stream and sent to the client browser which decodes the stream and displays the page to the user using the visual content and positional information.","The DOM tree and view tree and all state information (including Javascripts and cookies) for the client are stored on the storage device by the server-side browser.","Other aspects and advantages of the invention will become apparent from the following detailed description in combination with the accompanying drawings, illustrating, by way of example, the principles of the invention."],["BACKGROUND OF THE INVENTION 1.Technical Field The invention relates to Web browsing technology.","More particularly, the invention relates to a system and method for fast delivery of and fast rendering of Web pages on a lightweight network device.","2.Description of the Prior Art A typical Web page today contains not only HTML content but also, by reference to, other content elements such as graphics and external style sheets.","A browser acquires the referenced content elements by spawning worker threads that retrieve the data via multiple HTTP-TCP/IP connections.","This model works well for desktop systems containing large amounts of processing power and RAM.","However, it is usually not feasible for mobile, lightweight, embedded devices in which processing power and RAM are much limited.","The process for proper HTML parsing, validation, and layout code is normally quite complex and requires large amount of CPU and RAM to execute.","Size and manufacturing cost have been major concerns in providing embedded devices with a capacity to implement the process.","Although embedded devices are getting more and more processing power and RAM, the requirements of a proper browser engine still outdistance the resources available to even the highest end of embedded devices.","High levels of CPU usage are detrimental to the preservation of battery life, which is a very important consideration for today's mobile devices.","Heavy loads in HTTP communications, HTML parsing, HTML validation and layout may largely shorten the battery life.","Bandwidth usage is also a major constraint that must be considered in designing a browser model for embedded devices.","Text based HTML content, although compressible, is already quite large with respect to the limited transmission speed of today's wireless networks.","When image content designed for higher resolution and larger form factor display is included, one may quickly end up consuming a lot of wasted bandwidth transmitting redundant, not necessarily usable data to mobile devices.","Although various browsing technologies have been already presented in the marketplace, many of them fail to meet the tight constraints and stringent requirements of the embedded devices.","Writing a cross platform code base for these devices is an even bigger challenge, given the multitude of operating systems, memory models and processors available for embedded device manufacturers.","It would be advantageous to provide a server-based browser system that offloads heavy weight components of a traditional browser to a back end server.","It would further be advantageous to provide a server-based browser system that leaves a lightweight client device with a minimal task of implementing a customizable interface to display a pre-rendered representation of Web pages.","SUMMARY OF THE INVENTION The invention provides a server-based browser system.","The system offloads heavy weight components of a traditional browser to a back end server.","In addition, the invention leaves a lightweight client device with a minimal task of implementing a customizable interface to display a pre-rendered representation of Web pages.","A preferred embodiment of the invention provides a server-based browser and a client system browser.","The client browser determines the position of a user click on its display screen and sends a navigation request containing the coordinates of the dick or a hotspot ID, to the server-side browser.","The client browser will also accept a specific URL from a user and places the URL in the navigation request.","The server-side browser retrieves a Document Object Model (DOM) model and view tree from a storage device for the client It then finds the location on the Web page that the user clicked on using the coordinates or hotspot ID received from the client.","If there is a script associated with the location, it is executed and the resulting page location is requested from the appropriate server.","If there is a URL associated with the location, it is requested from the appropriate server.","The response Web page HTML page definition is parsed and a DOM tree model is created.","The server-side browser uses the DOM tree and creates a two-dimensional view tree model, and stores both trees on the storage device.","The server-side browser retrieves a style sheet, layout algorithms, and device constraints for the client device from the storage device.","It lays out the view model using the style sheet, layout algorithms, and device constraints onto a virtual page.","If the client browser is a subset of the server-side browser engine, then the DOM tree and view tree are compressed into a stream and sent to the client browser which decodes the stream, translates the DOM tree and view tree, and displays the page to the user.","Otherwise, the server-side browser determines the visual content on the virtual page and the location of the visual content.","Visual content and positional information are highly compressed and formatted into a stream and sent to the client browser which decodes the stream and displays the page to the user using the visual content and positional information.","The DOM tree and view tree and all state information (including Javascripts and cookies) for the client are stored on the storage device by the server-side browser.","Other aspects and advantages of the invention will become apparent from the following detailed description in combination with the accompanying drawings, illustrating, by way of example, the principles of the invention.","BRIEF DESCRIPTION OF THE DRAWINGS FIG.","1 is a block schematic diagram of a preferred embodiment of the invention showing a server-side browser and a client browser according to the invention; FIG.","2 is a block schematic diagram of a server-side browser and renderer and client device according to the invention; FIG.","3 is a block schematic diagram of a server-side browser engine and client device according to the invention; FIG.","4 is a block schematic diagram showing an HTML document tree according to the invention; FIG.","5 is a block schematic diagram showing a view tree according to the invention; FIG.","6 is a block schematic diagram of a view layout of a Web page according to the invention; FIG.","7 is a block schematic diagram of a flowchart of an embodiment of the invention according to the invention; and FIG.","8 is a block schematic diagram of a task viewpoint of a preferred embodiment of the invention according to the invention.","DETAILED DESCRIPTION OF THE INVENTION The invention is embodied in a server-based browser system.","A system according to the invention offloads heavy weight components of a traditional browser to a back end server.","In addition, the invention provides a server-based browser system that leaves a lightweight client device with a minimal task of implementing a customizable interface to display a pre-rendered representation of Web pages.","A typical desktop browser requires a large amount of processor time to handle the load of obtaining Web page information and parsing the HTML code in the page.","Secondary requests within the HTML code also require a large amount of processing time.","Simple client devices such as PDAs, cellphones, and low CPU power set top boxes cannot handle the amount of processing power required to perform typical Web browsing.","Additionally, bandwidth on devices such as cellphones are limited and are charged to the user on a byte basis.","This means that the large data transfers involved in typical Web browsing are too expensive in terms of bandwidth limitations and cost to the user.","Referring to FIG.","1, the invention provides a server-based 101 browser 104 that converts Web pages to a minimal format adaptable to different types of wired/wireless client devices such as cellphones, PDAs, and set top TV boxes.","A light weight browser 105 is provided on a client device 102 to accept highly compressed streams of data from the server-side browser 104.The server-side browser 104 accesses information across the Internet or intranet 103.With respect to FIG.","2, a user using a client device 206, 207 requests a Web page from a light weight browser or the light weight browser itself requests a Web page.","The request is sent from the client 206, 207 to a server-side browser 201.The server-side browser 201 identifies the Web page that the user requested and sends a request to the proper server across the Internet or intranet.","When the requested Web page is returned to the server-side browser 201, the server-side browser 201 loads the initial HTML document.","The received HTML document consists of text, which comprises the content of the document, and tags, which define the structure and appearance of the document.","The server-side browser 201 looks at any dependencies within the document (e.g., referenced images, Javascript files that need to be loaded remotely, etc.)","and opens multiple sockets to get the required information across the Internet or intranet.","The server-side browser 201 gathers the Web page information and lays out the information to calculate how to display the information on a particular device.","The server-side browser 201 makes a second pass at the layout information and then transcodes the information into a very tight data stream that can be represented on the client 206, 207.The stream includes positional information that is not present in the normal HTML for the Web page.","The server-side browser 201 takes all of the HTML information for a Web page and represents it as two dimensional content on the server.","This two-dimensional representation helps the browser determine the exact position information positional information for all of the pieces of a particular HTML Web page.","If there are any images or other dependencies, the server-side browser 201 scales (or crops) the images down and/or preprocesses the images (e.g., reduce the bit depth of an image, dither the image, etc.)","to match the capabilities of the client 206, 207.The browser then creates a compressed stream that is sent down to the client.","This compressed stream contains text and images with exact positional information (that tells where the pieces of content fit on the page), and hot spot information.","The browser maps each rectangular hotspot region to a related URL.","Thus, no extraneous information, such as hyperlink URLs, or Cookie Data is sent down to the client.","The server-side browser 205 uses device characteristics information 202 for each supported device type, consisting of the associated default CSS style sheet, a list of supported fonts and their text metrics, device aspect ratios, pixel depth, color palette, form control dimensions, preferred layout algorithms, a DTD specifying level of HTML compliance, JavaScript policies, the User-Agent String to represent to a Web Server, and the compressed stream format encoder, to properly determine the ideal representation of Web content on the device.","Various examples of potential clients with vastly different device characteristics could include: a cellphone 206 with a minimal CPU and minimal RAM; a PDA client with a small display but powerful CPU and high RAM; or a TV box 207 with a large display, high capacity CPU, low RAM, and non TCP/IP based communications.","The server-side browser 205 utilizes many different layout algorithms to properly render information with these various classes of clients that all have different: display, CPU, RAM, and 10 constraints.","A lightweight client browser is provided on the clients 206, 207 that accepts a standard information format from the server-side browser 201 and displays the data streams.","This means that the server-side browser 201 can be updated at any time without affecting the clients 206, 207.Referring to FIGS.","3, 4, and 5, a preferred embodiment of the invention provides a server-side browser engine 302 that accepts HTML (e.g., HTML 4.0 content with support for CSS version 1) pages 303 and parses the HTML 304.The parser 304 creates a Document Object Model (DOM) object hierarchy model 305.The DOM model 305 is a tree representation of the HTML in the Web page, which is well known in the art.","A style sheet 306 tells the server-side browser 301 what fonts to use for standard HTML tags (e.g., bold), border sizes, and other stylistic information for each supported client device.","A device information file 307 is also used by the browser that tells the server-side browser 301 the limitations of a client device (e.g., the size of a text control, fonts available, aspect ratio, etc.).","The server-side browser 301 discovers what device type the client is when the client signs onto the server.","Using the style sheet 306 and the device information 307 for the particular device, the server-side browser 301 creates a view tree 308 which is a two-dimensional model of the parent-child relationship between viewable objects of the DOM model 305.A DOM tree is shown in FIG.","4.The HTML 401 is the root of the tree and underneath has a Body tag 402.Under the Body tag 402 is an Anchor tag 403 which has Text 404 underneath.","There is also a Table 405 beneath the Anchor tag 403.The Table 405 has a row 406 which has a cell 407.The browser makes views of DOM tree elements that are visible.","For example, a Text view 408 is made of the Text 404, a Table view 409 is made of the Table 405, a TR view 410 is made of the row 406, and a TD view 411 is made of the cell 407.An exemplary view model is shown in FIG.","5, where a Body view 501 is the root of the tree.","A Table view 502 sits below the Body view 501 and has a TR view 503 below it.","A TD view 504 is below the TR view 503.The TR view 504 has both a text view 505 and an image view 506 below it.","The server-side browser 301 then lays out the view model 308 on a virtual page.","Each view in the view model 308 has a rectangular area of information attached to it.","The server-side browser 301 lays out the views using specific layout algorithms for each view (e.g., tables, block levels, etc.)","and uses the style sheet 306 and device information 307 for the particular device to make sure that any special characteristics or behaviors of the device are taken into account For example, for a TV box client, there may be a fixed table width defined in the view and fixed table widths are ignored by the TV box, or, one pixel width table borders are only supported on the TV box client.","An example of a layout algorithm is every cell in a table is considered a row in itself, for a certain device each cell is to be laid out vertically instead of the normal horizontal layout.","With respect to FIG.","6, the server-side browser performs the first layout pass which lays out all of the views of the page 601.The page 601 is a virtual view of how the page looks.","In this example, a body view 605 has a table view 604 within it.","The Table view 604 has a row 603 which has a cell with text in it 602.Each view has a rectangular area that it represents and a parent-child relationship.","This allows the server-side browser to walk through the viewable area to see what it represents.","A second pass is made to determine what the real content is on the page.","HTML authors often define multiply nested tables to constrain a layout of an HTML page.","The server-side browser throws away the unnecessary information that does not contribute to the appearance of the page (e.g., not renderable).","The server-side browser is concerned with the physical positioning of content elements within the page.","For example, the browser finds text 602 on the page 601 and accumulates the text into a string and compresses it to send to the client.","The server-side browser also sends the client information that tells the client where the text should be displayed in its viewing area.","This information is placed in a binary format that the client device feeds into.","The DOM tree and view tree and all state information (including Javascripts) for the client are preserved on the server.","This is to ensure that, when the user is interacting with a page on the client, the server-side browser knows where the user has clicked on the page.","The server-side browser does not send URL links to the client.","This means that the client only has positional or hotspot information relating to where the user clicked or pressed on the page and sends that coordinate or hotspot information to the server-side browser.","The server-side browser looks at the DOM tree and view model to find where the coordinates map onto the page.","The view model is used to find the location of the coordinates or hotspot ID and the DOM tree is used to find the URL or script associated with the clicked area.","When the server-side browser finds the area that corresponds to location where the user clicked, the server-side browser executes whatever script or URL request that corresponds with the area.","The server-side browser then updates the client with the new page if the action was navigational, for example.","Cookie data are impractical to place on the client device.","The server-side browser caches all of the user's cookies in a user state that the server-side browser preserves on the server for each user.","A session object exists on both the server and the client The session protocol is used by the client and server to communicate information.","For example, when the client first signs onto the server, it tells the server its configuration information and other information.","The server-side browser looks into its database and finds the client device's style and device information.","Referring to FIG.","7, a flowchart of an another preferred embodiment of the invention is shown.","The client browser 711 sends a navigation request to the server-side browser engine 701.The browser session 702 retrieves the client specific style sheet 703 and layout algorithms 704.The browser session 702 fetches the requested Web page from the Internet or intranet.","The HTML Web page 705 is parsed and a DOM tree model is created 706.The DOM tree model is then used to create a two-dimensional view tree model 707.The invention lays out the view tree based on the client specific style sheet and layout algorithms 708.Any additional layouts are performed on the view tree to fit the client device constraints 709.The resulting two-dimensional view tree and DOM tree are encoded in a lightweight format and sent to the client which decodes the format 710 and displays the resulting page to the user through the client browser 711.With respect to FIG.","8, a task viewpoint of the invention is shown.","The Client Browser module 812 determines the position of a user dick or press on its display screen and sends a navigation request containing the coordinates of the click or press, or the hotspot ID, to the server-side browser through the Client Session module 811 to the Server Session module 804.The Client Browser module 812 will also accept a specific URL from a user and places the URL in the navigation request.","The Server Session module 804 forwards the navigation request to the Find Position module 805.The Find Position module 805 accepts Specific URLs from the client, coordinates of a user's click or press, and hotspot IDs 810.The Find Position module 805 forwards any specific URL to the Web Page Manager 902.Otherwise, the Find Position module 805 retrieves the DOM model and view tree from the storage device 807 for the particular client.","It then finds the location on the Web page that the user clicked or pressed on using the coordinates or hotspot ID received from the client.","If there is a script associated with the location, it is executed and the resulting page location is sent to the Web Page Manager 902.If there is a URL associated with the location, it is forwarded to the Web Page Manager 902.The Web Page Manager 802 receives the URL from the Find Position module 805 and formats a request for the Web page.","The request is sent to the W e b Session module 803.The Web page is returned to the Web Session module 803 and is forwarded to the Web Page Manager 802.The Web page Manager 802 forwards the HTML Web page definition to the Parse HTML module 801.The Parse HTML module 801 parses the HTML and creates a DOM tree model which it stores on the storage device 807.The Web page Manager 802 then activates the Create View module 806.The Create View module 806 uses the DOM model stored on the storage device 807, creates a two-dimensional view tree model, and stores it on the storage device 807.Once the view tree is completed, the Web page Manager 802 retrieves the style sheet, layout algorithms, and device constraints for the specific client device are retrieved from the storage device 807.The Web page Manager 802 lays out the view model using the style sheet, layout algorithms, and device constraints.","If the Client Browser 812 is a subset of the server-side browser engine, then the DOM tree and view tree are compressed into a stream 809 and sent through the Server Session module 804 to the Client Browser 812 via the Client Session module 811.The Client Browser 812 decodes the stream, translates the DOM tree and view tree, and displays the page to the user.","Otherwise, the Web page Manager 802 performs the first layout pass and lays out all of the views of the page.","The Web page Manager 802 then makes a second pass to determine the visual content on the page.","Visual content and positional information are highly compressed and formatted into a stream 809 and forwarded to the Server Session module 804.The Server Session module 804 sends the stream to the Client Browser 812 via the Client Session module 811.The Client Browser 812 decodes the stream and displays the page to the user using the visual content and positional information.","The DOM tree and view tree and all state information (including Javascripts and cookies) for the client are stored on the storage device 807 by the Web page Manager 802.Although the invention is described herein with reference to the preferred embodiment, one skilled in the art will readily appreciate that other applications may be substituted for those set forth herein without departing from the spirit and scope of the present invention.","Accordingly, the invention should only be limited by the claims included below."]],"string":"[\n [\n \"Server-based browser system\",\n \"A server-based browser system provides a server-based browser and a client system browser.\",\n \"The client browser reports the position of a user click on its display screen, hotspot ID, or URL to the server-side browser which retrieves a Document Object Model (DOM) model and view tree for the client and finds the location on the Web page that the user clicked on using the coordinates or hotspot ID received from the client.\",\n \"If there is a script associated with the location, it is executed and the resulting page location is requested from the appropriate server.\",\n \"If there is a URL associated with the location, it is requested from the appropriate server.\",\n \"The response Web page HTML definition is parsed and a DOM tree model is created which is used to create a view tree model.\",\n \"The server-side browser retrieves a style sheet, layout algorithms, and device constraints for the client device and lays out the view model using them onto a virtual page and determines the visual content.\",\n \"Textual and positional information are highly compressed and formatted into a stream and sent to the client browser which decodes the stream and displays the page to the user using the textual and positional information.\"\n ],\n [\n \"1.A process for providing a server-based Web browser that interacts with a plurality of client device types, comprising the steps of: (a) providing browsing means on a server for accepting a Web page from a Web server, said browsing means comprising Web page layout means for laying out said Web page in a two-dimensional virtual page, said Web page layout means determining what information is visible on said virtual page, said Web page layout means calculating positioning of said visible information within said Web page, and said browsing means creating a compressed stream containing said visible information and the positioning information of said visible information; (b) providing client browsing means on a client system for accepting compressed streams representing display pages; and (c) providing means on said server for sending said compressed stream to said client browsing means; wherein said client browsing means decompresses said compressed stream and displays said visible information using said positioning information.\",\n \"2.The process of claim 1, wherein said browsing means comprises: a database of style sheets on a storage device for supported client devices; a database of layout algorithms on said storage device for supported client devices; and a database of device constraint information on said storage device for supported client devices; wherein said Web page layout means retrieves a style sheet, layout algorithms, and device constraints from said storage device for said client system; and wherein said Web page layout means lays out said Web page based on said client system's style sheet, layout algorithms, and device constraints.\",\n \"3.The process of claim 1, wherein said step of providing browsing means further comprises the sub-steps of: parsing said Web page's HTML code to create a Document Object Model (DOM) tree; creating a view tree using said DOM tree; and storing said DOM tree and said view tree on a storage device for said client system.\",\n \"4.The process of claim 3, wherein said client browsing means accepts a URL from a user or determines a position or hotspot ID of a user's click or press on a display page, wherein said client browsing means sends said URL, said position, or said hotspot ID to said browsing means, and wherein said browsing means accepts said URL, said position, or said hotspot ID from said client browsing means.\",\n \"5.The process of claim 4, wherein said step of providing browsing means further comprises the sub-steps of: retrieving a DOM tree and view tree from said storage device for said client system; determining the location of said position or hotspot ID on said Web page using said view tree; finding a script or URL associated with the location using said DOM tree; wherein if a script is associated with the location then executing the script and determining the resulting URL; and requesting the Web page associated with said resulting URL or said associated URL from the appropriate server.\",\n \"6.The process of claim 3, wherein said step of providing browsing means further comprises the sub-step of: saving said client system's state information on said storage device for a particular user; wherein said client system's state information includes Javascripts and cookies; and wherein said client system's state information allows said browsing means to track the current state of said client system and the particular user.\",\n \"7.A process for providing a cross platform Web browser that interacts with a plurality of client device types, comprising the steps of: (a) providing browsing means on a server for accepting a Web page from a Web server; said step pf providing browsing means further comprising the sub-steps of: parsing said Web page's HTML code to create a Document Object Model (DOM) tree; creating a view tree using said DOM tree; providing Web page layout means for laying out said Web page in a two-dimensional virtual page; wherein said Web page layout means determines what information is visible on said virtual page; wherein said Web page layout means removes unnecessary structural information and retains visible information in said view tree; and storing said DOM tree and said view tree on a storage device for said client system; wherein said browsing means creates a compressed stream containing said DOM tree and said view tree; (b) providing client browsing means on a client system for accepting compressed streams; and (c) providing means on said server for sending said compressed stream to said client browsing means; wherein said client browsing means decompresses said compressed stream and displays said visible information using said DOM tree and said view tree.\",\n \"8.The process of claim 7, wherein said browsing means comprises: a database of style sheets on said storage device for supported client devices; a database of layout algorithms on said storage device for supported client devices; and a database of device constraint information on said storage device for supported client devices; wherein said Web page layout means retrieves a style sheet, layout algorithms, and device constraints from said storage device for said client system; and wherein said Web page layout means lays out said Web page based on said client system's style sheet, layout algorithms, and device constraints.\",\n \"9.The process of claim 7, wherein said client browsing means accepts a URL from a user or determines a position or hotspot ID of a user's click or press on a display page, wherein said client browsing means sends said URL, said position, or said hotspot ID to said browsing means, and wherein said browsing means accepts said URL, said position, or said hotspot ID from said client browsing means.\",\n \"10.The process of claim 9, wherein said step of providing browsing means further comprises the sub-steps of: retrieving a DOM tree and view tree from said storage device for said client system; determining the location of said position or hotspot ID on said Web page using said view tree; and finding a script or URL associated with the location using said DOM tree; wherein if a script is associated with the location then executing the script and determining the resulting URL; and requesting the Web page associated with said resulting URL or said associated URL from the appropriate server.\",\n \"11.The process of claim 7, wherein said step of providing browsing means further comprising the sub-step of: saving said client system's state information on said storage device for a particular user; wherein said client system's state information includes Javascripts and cookies; and wherein said client system's state information allows said browsing means to track the current state of said client system and the particular user.\",\n \"12.An apparatus for a server-based Web browser that interacts with a plurality of client device types, comprising: (a) browsing means on a server for accepting a Web page from a Web server, said browsing means further comprising Web page layout means for laying out said Web page in a two-dimensional virtual page, said Web page layout means determining what information is visible on said virtual page, and said Web page layout means calculating positioning of said visible information within said Web page; wherein said browsing means creates a compressed stream containing said visible information and the positioning information of said visible information; (b) client browsing means on a client system for accepting compressed streams representing display pages; and (c) means on said server for sending said compressed stream to said client browsing means; wherein said client browsing means decompresses said compressed stream and displays said visible information using said positioning information.\",\n \"13.The apparatus of claim 12, wherein said browsing means further comprises: a database of style sheets on a storage device for supported client devices; a database of layout algorithms on said storage device for supported client devices; and a database of device constraint information on said storage device for supported client devices; wherein said Web page layout means retrieves a style sheet, layout algorithms, and device constraints from said storage device for said client system; and wherein said Web page layout means lays out said Web page based on said client system's style sheet, layout algorithms, and device constraints.\",\n \"14.The apparatus of claim 12, wherein said browsing means further comprises: a module for parsing said Web page's HTML code to create a Document Object Model (DOM) tree; a module for creating a view tree using said DOM tree; and a module for storing said DOM tree and said view tree on a storage device for said client system.\",\n \"15.The apparatus of claim 14, wherein said client browsing means accepts a URL from a user or determines a position or hotspot ID of a user's click or press on a display page, wherein said client browsing means sends said URL, said position, or said hotspot ID to said browsing means, and wherein said browsing means accepts said URL, said position, or said hotspot ID from said client browsing means.\",\n \"16.The apparatus of claim 15, wherein said browsing means further comprises: a module for retrieving a DOM tree and view tree from said storage device for said client system; a module for determining the location of said position or hotspot ID on said Web page using said view tree; a module for finding a script or URL associated with the location using said DOM tree; wherein if a script is associated with the location then executing the script and determining the resulting URL; and a module for requesting the Web page associated with said resulting URL or said associated URL from the appropriate server.\",\n \"17.The apparatus of claim 14, wherein said browsing means further comprises: a module for saving said client system's state information on said storage device for a particular user; wherein said client system's state information includes Javascripts and cookies; and wherein said client system's state information allows said browsing means to track the current state of said client system and the particular user.\",\n \"18.An apparatus for a cross platform Web browser that interacts with a plurality of client device types, comprising: (a) browsing means on a server for accepting a Web page from a Web server; said browsing means further comprising: a module for parsing said Web page's HTML code to create a Document Object Model (DOM) tree; a module for creating a view tree using said DOM tree; Web page layout means for laying out said Web page in a two-dimensional virtual page; wherein said Web page layout means determines what information is visible on said virtual page; wherein said Web page layout means removes unnecessary structural information and retains visible information in said view tree; and a module for storing said DOM tree and said view tree on a storage device for said client system; wherein said browsing means creates a compressed stream containing said DOM tree and said view tree; (b) client browsing means on a client system for accepting compressed streams; (c) means on said server for sending said compressed stream to said client browsing means; and wherein said client browsing means decompresses said compressed stream and displays said visible information using said DOM tree and said view tree.\",\n \"19.The apparatus of claim 18, wherein said browsing means further comprises: a database of style sheets on said storage device for supported client devices; a database of layout algorithms on said storage device for supported client devices; a database of device constraint information on said storage device for supported client devices; wherein said Web page layout means retrieves a style sheet, layout algorithms, and device constraints from said storage device for said client system; and wherein said Web page layout means lays out said Web page based on said client system's style sheet, layout algorithms, and device constraints.\",\n \"20.The apparatus of claim 18, wherein said client browsing means accepts a URL from a user or determines a position or hotspot ID of a user's click or press on a display page, wherein said client browsing means sends said URL, said position, or said hotspot ID to said browsing means, and wherein said browsing means accepts said URL, said position, or said hotspot ID from said client browsing means.\",\n \"21.The apparatus of claim 20, wherein said browsing means further comprises: a module for retrieving a DOM tree and view tree from said storage device for said client system; a module for determining the location of said position or hotspot ID on said Web page using said view tree; a module for finding a script or URL associated with the location using said DOM tree; wherein if a script is associated with the location then executing the script and determining the resulting URL; and a module for requesting the Web page associated with said resulting URL or said associated URL from the appropriate server.\",\n \"22.The apparatus of claim 18, wherein said browsing means further comprises: a module for saving said client system's state information on said storage device for a particular user; wherein said client system's state information includes Javascripts and cookies; and wherein said client system's state information allows said browsing means to track the current state of said client system and the particular user.\",\n \"23.A program storage medium readable by a computer, tangibly embodying a program of instructions executable by the computer to perform method steps for a server-based Web browser that interacts with a plurality of client device types, comprising the steps of: (a) providing browsing means on a server for accepting a Web page from a Web server, said browsing means providing Web page layout means for laying out said Web page in a two-dimensional virtual page, said Web page layout means determining what information is visible on said virtual page, said Web page layout means calculating positioning of said visible information within said Web page; and said browsing means creating a compressed stream containing said visible information and the positioning information of said visible information; (b) providing client browsing means on a client system for accepting compressed streams representing display pages; and (c) providing means on said server for sending said compressed stream to said client browsing means; wherein said client browsing means decompresses said compressed stream and displays said visible information using said positioning information.\",\n \"24.The program storage medium of claim 23, wherein said browsing means comprises: a database of style sheets on a storage device for supported client devices; a database of layout algorithms on said storage device for supported client devices; and a database of device constraint information on said storage device for supported client devices; wherein said Web page layout means retrieves a style sheet, layout algorithms, and device constraints from said storage device for said client system; and wherein said Web page layout means lays out said Web page based on said client system's style sheet, layout algorithms, and device constraints.\",\n \"25.The program storage medium of claim 23, wherein said step of providing browsing means further comprising the sub-steps of: parsing said Web page's HTML code to create a Document Object Model (DOM) tree; creating a view tree using said DOM tree; and storing said DOM tree and said view tree on a storage device for said client system.\",\n \"26.The program storage medium of claim 25, wherein said client browsing means accepts a URL from a user or determines a position or hotspot ID of a user's click or press on a display page, wherein said client browsing means sends said URL, said position, or said hotspot ID to said browsing means, and wherein said browsing means accepts said URL, said position, or said hotspot ID from said client browsing means.\",\n \"27.The program storage medium of claim 26, wherein said step of providing browsing means further comprising the sub-steps of: retrieving a DOM tree and view tree from said storage device for said client system; determining the location of said position or hotspot ID on said Web page using said view tree; finding a script or URL associated with the location using said DOM tree; wherein if a script is associated with the location then executing the script and determining the resulting URL; and requesting the Web page associated with said resulting URL or said associated URL from the appropriate server.\",\n \"28.The program storage medium of claim 23, wherein said step of providing browsing means further comprising the step of: saving said client system's state information on said storage device for a particular user; wherein said client system's state information includes Javascripts and cookies; and wherein said client system's state information allows said browsing means to track the current state of said client system and the particular user.\",\n \"29.A program storage medium readable by a computer, tangibly embodying a program of instructions executable by the computer to perform method steps for a cross platform Web browser that interacts with a plurality of client device types, comprising the steps of: (a) providing browsing means on a server for accepting a Web page from a Web server; said step of providing browsing means further comprising the sub-steps of: parsing said Web page's HTML code to create a Document Object Model (DOM) tree; creating a view tree using said DOM tree; providing Web page layout means for laying out said Web page in a two-dimensional virtual page; wherein said Web page layout means determines what information is visible on said virtual page; wherein said Web page layout means removes unnecessary structural information and retains visible information in said view tree; and storing said DOM tree and said view tree on a storage device for said client system; wherein said browsing means creates a compressed stream containing said DOM tree and said view tree; (b) providing client browsing means on a client system for accepting compressed streams; and (c) providing means on said server for sending said compressed stream to said client browsing means; wherein said client browsing means decompresses said compressed stream and displays said visible information using said DOM tree and said view tree.\",\n \"30.The program storage medium of claim 29, wherein said browsing means comprises: a database of style sheets on said storage device for supported client devices; a database of layout algorithms on said storage device for supported client devices; and a database of device constraint information on said storage device for supported client devices; wherein said Web page layout means retrieves a style sheet, layout algorithms, and device constraints from said storage device for said client system; and wherein said Web page layout means lays out said Web page based on said client system's style sheet, layout algorithms, and device constraints.\",\n \"31.The program storage medium of claim 29, wherein said client browsing means accepts a URL from a user or determines a position or hotspot ID of a user's click or press on a display page, wherein said client browsing means sends said URL, said position, or said hotspot ID to said browsing means, and wherein said browsing means accepts said URL, said position, or said hotspot ID from said client browsing means.\",\n \"32.The program storage medium of claim 31, wherein said step of providing browsing means further comprising the steps of: retrieving a DOM tree and view tree from said storage device for said client system; determining the location of said position or hotspot ID on said Web page using said view tree; finding a script or URL associated with the location using said DOM tree; wherein if a script is associated with the location then executing the script and determining the resulting URL; and requesting the Web page associated with said resulting URL or said associated URL from the appropriate server.\",\n \"33.The program storage medium of claim 29, wherein said step of providing browsing means further comprises the sub-step of: saving said client system's state information on said storage device for a particular user; wherein said client system's state information includes Javascripts and cookies; and wherein said client system's state information allows said browsing means to track the current state of said client system and the particular user.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>1.Technical Field The invention relates to Web browsing technology.\",\n \"More particularly, the invention relates to a system and method for fast delivery of and fast rendering of Web pages on a lightweight network device.\",\n \"2.Description of the Prior Art A typical Web page today contains not only HTML content but also, by reference to, other content elements such as graphics and external style sheets.\",\n \"A browser acquires the referenced content elements by spawning worker threads that retrieve the data via multiple HTTP-TCP/IP connections.\",\n \"This model works well for desktop systems containing large amounts of processing power and RAM.\",\n \"However, it is usually not feasible for mobile, lightweight, embedded devices in which processing power and RAM are much limited.\",\n \"The process for proper HTML parsing, validation, and layout code is normally quite complex and requires large amount of CPU and RAM to execute.\",\n \"Size and manufacturing cost have been major concerns in providing embedded devices with a capacity to implement the process.\",\n \"Although embedded devices are getting more and more processing power and RAM, the requirements of a proper browser engine still outdistance the resources available to even the highest end of embedded devices.\",\n \"High levels of CPU usage are detrimental to the preservation of battery life, which is a very important consideration for today's mobile devices.\",\n \"Heavy loads in HTTP communications, HTML parsing, HTML validation and layout may largely shorten the battery life.\",\n \"Bandwidth usage is also a major constraint that must be considered in designing a browser model for embedded devices.\",\n \"Text based HTML content, although compressible, is already quite large with respect to the limited transmission speed of today's wireless networks.\",\n \"When image content designed for higher resolution and larger form factor display is included, one may quickly end up consuming a lot of wasted bandwidth transmitting redundant, not necessarily usable data to mobile devices.\",\n \"Although various browsing technologies have been already presented in the marketplace, many of them fail to meet the tight constraints and stringent requirements of the embedded devices.\",\n \"Writing a cross platform code base for these devices is an even bigger challenge, given the multitude of operating systems, memory models and processors available for embedded device manufacturers.\",\n \"It would be advantageous to provide a server-based browser system that offloads heavy weight components of a traditional browser to a back end server.\",\n \"It would further be advantageous to provide a server-based browser system that leaves a lightweight client device with a minimal task of implementing a customizable interface to display a pre-rendered representation of Web pages.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>The invention provides a server-based browser system.\",\n \"The system offloads heavy weight components of a traditional browser to a back end server.\",\n \"In addition, the invention leaves a lightweight client device with a minimal task of implementing a customizable interface to display a pre-rendered representation of Web pages.\",\n \"A preferred embodiment of the invention provides a server-based browser and a client system browser.\",\n \"The client browser determines the position of a user click on its display screen and sends a navigation request containing the coordinates of the dick or a hotspot ID, to the server-side browser.\",\n \"The client browser will also accept a specific URL from a user and places the URL in the navigation request.\",\n \"The server-side browser retrieves a Document Object Model (DOM) model and view tree from a storage device for the client It then finds the location on the Web page that the user clicked on using the coordinates or hotspot ID received from the client.\",\n \"If there is a script associated with the location, it is executed and the resulting page location is requested from the appropriate server.\",\n \"If there is a URL associated with the location, it is requested from the appropriate server.\",\n \"The response Web page HTML page definition is parsed and a DOM tree model is created.\",\n \"The server-side browser uses the DOM tree and creates a two-dimensional view tree model, and stores both trees on the storage device.\",\n \"The server-side browser retrieves a style sheet, layout algorithms, and device constraints for the client device from the storage device.\",\n \"It lays out the view model using the style sheet, layout algorithms, and device constraints onto a virtual page.\",\n \"If the client browser is a subset of the server-side browser engine, then the DOM tree and view tree are compressed into a stream and sent to the client browser which decodes the stream, translates the DOM tree and view tree, and displays the page to the user.\",\n \"Otherwise, the server-side browser determines the visual content on the virtual page and the location of the visual content.\",\n \"Visual content and positional information are highly compressed and formatted into a stream and sent to the client browser which decodes the stream and displays the page to the user using the visual content and positional information.\",\n \"The DOM tree and view tree and all state information (including Javascripts and cookies) for the client are stored on the storage device by the server-side browser.\",\n \"Other aspects and advantages of the invention will become apparent from the following detailed description in combination with the accompanying drawings, illustrating, by way of example, the principles of the invention.\"\n ],\n [\n \"BACKGROUND OF THE INVENTION 1.Technical Field The invention relates to Web browsing technology.\",\n \"More particularly, the invention relates to a system and method for fast delivery of and fast rendering of Web pages on a lightweight network device.\",\n \"2.Description of the Prior Art A typical Web page today contains not only HTML content but also, by reference to, other content elements such as graphics and external style sheets.\",\n \"A browser acquires the referenced content elements by spawning worker threads that retrieve the data via multiple HTTP-TCP/IP connections.\",\n \"This model works well for desktop systems containing large amounts of processing power and RAM.\",\n \"However, it is usually not feasible for mobile, lightweight, embedded devices in which processing power and RAM are much limited.\",\n \"The process for proper HTML parsing, validation, and layout code is normally quite complex and requires large amount of CPU and RAM to execute.\",\n \"Size and manufacturing cost have been major concerns in providing embedded devices with a capacity to implement the process.\",\n \"Although embedded devices are getting more and more processing power and RAM, the requirements of a proper browser engine still outdistance the resources available to even the highest end of embedded devices.\",\n \"High levels of CPU usage are detrimental to the preservation of battery life, which is a very important consideration for today's mobile devices.\",\n \"Heavy loads in HTTP communications, HTML parsing, HTML validation and layout may largely shorten the battery life.\",\n \"Bandwidth usage is also a major constraint that must be considered in designing a browser model for embedded devices.\",\n \"Text based HTML content, although compressible, is already quite large with respect to the limited transmission speed of today's wireless networks.\",\n \"When image content designed for higher resolution and larger form factor display is included, one may quickly end up consuming a lot of wasted bandwidth transmitting redundant, not necessarily usable data to mobile devices.\",\n \"Although various browsing technologies have been already presented in the marketplace, many of them fail to meet the tight constraints and stringent requirements of the embedded devices.\",\n \"Writing a cross platform code base for these devices is an even bigger challenge, given the multitude of operating systems, memory models and processors available for embedded device manufacturers.\",\n \"It would be advantageous to provide a server-based browser system that offloads heavy weight components of a traditional browser to a back end server.\",\n \"It would further be advantageous to provide a server-based browser system that leaves a lightweight client device with a minimal task of implementing a customizable interface to display a pre-rendered representation of Web pages.\",\n \"SUMMARY OF THE INVENTION The invention provides a server-based browser system.\",\n \"The system offloads heavy weight components of a traditional browser to a back end server.\",\n \"In addition, the invention leaves a lightweight client device with a minimal task of implementing a customizable interface to display a pre-rendered representation of Web pages.\",\n \"A preferred embodiment of the invention provides a server-based browser and a client system browser.\",\n \"The client browser determines the position of a user click on its display screen and sends a navigation request containing the coordinates of the dick or a hotspot ID, to the server-side browser.\",\n \"The client browser will also accept a specific URL from a user and places the URL in the navigation request.\",\n \"The server-side browser retrieves a Document Object Model (DOM) model and view tree from a storage device for the client It then finds the location on the Web page that the user clicked on using the coordinates or hotspot ID received from the client.\",\n \"If there is a script associated with the location, it is executed and the resulting page location is requested from the appropriate server.\",\n \"If there is a URL associated with the location, it is requested from the appropriate server.\",\n \"The response Web page HTML page definition is parsed and a DOM tree model is created.\",\n \"The server-side browser uses the DOM tree and creates a two-dimensional view tree model, and stores both trees on the storage device.\",\n \"The server-side browser retrieves a style sheet, layout algorithms, and device constraints for the client device from the storage device.\",\n \"It lays out the view model using the style sheet, layout algorithms, and device constraints onto a virtual page.\",\n \"If the client browser is a subset of the server-side browser engine, then the DOM tree and view tree are compressed into a stream and sent to the client browser which decodes the stream, translates the DOM tree and view tree, and displays the page to the user.\",\n \"Otherwise, the server-side browser determines the visual content on the virtual page and the location of the visual content.\",\n \"Visual content and positional information are highly compressed and formatted into a stream and sent to the client browser which decodes the stream and displays the page to the user using the visual content and positional information.\",\n \"The DOM tree and view tree and all state information (including Javascripts and cookies) for the client are stored on the storage device by the server-side browser.\",\n \"Other aspects and advantages of the invention will become apparent from the following detailed description in combination with the accompanying drawings, illustrating, by way of example, the principles of the invention.\",\n \"BRIEF DESCRIPTION OF THE DRAWINGS FIG.\",\n \"1 is a block schematic diagram of a preferred embodiment of the invention showing a server-side browser and a client browser according to the invention; FIG.\",\n \"2 is a block schematic diagram of a server-side browser and renderer and client device according to the invention; FIG.\",\n \"3 is a block schematic diagram of a server-side browser engine and client device according to the invention; FIG.\",\n \"4 is a block schematic diagram showing an HTML document tree according to the invention; FIG.\",\n \"5 is a block schematic diagram showing a view tree according to the invention; FIG.\",\n \"6 is a block schematic diagram of a view layout of a Web page according to the invention; FIG.\",\n \"7 is a block schematic diagram of a flowchart of an embodiment of the invention according to the invention; and FIG.\",\n \"8 is a block schematic diagram of a task viewpoint of a preferred embodiment of the invention according to the invention.\",\n \"DETAILED DESCRIPTION OF THE INVENTION The invention is embodied in a server-based browser system.\",\n \"A system according to the invention offloads heavy weight components of a traditional browser to a back end server.\",\n \"In addition, the invention provides a server-based browser system that leaves a lightweight client device with a minimal task of implementing a customizable interface to display a pre-rendered representation of Web pages.\",\n \"A typical desktop browser requires a large amount of processor time to handle the load of obtaining Web page information and parsing the HTML code in the page.\",\n \"Secondary requests within the HTML code also require a large amount of processing time.\",\n \"Simple client devices such as PDAs, cellphones, and low CPU power set top boxes cannot handle the amount of processing power required to perform typical Web browsing.\",\n \"Additionally, bandwidth on devices such as cellphones are limited and are charged to the user on a byte basis.\",\n \"This means that the large data transfers involved in typical Web browsing are too expensive in terms of bandwidth limitations and cost to the user.\",\n \"Referring to FIG.\",\n \"1, the invention provides a server-based 101 browser 104 that converts Web pages to a minimal format adaptable to different types of wired/wireless client devices such as cellphones, PDAs, and set top TV boxes.\",\n \"A light weight browser 105 is provided on a client device 102 to accept highly compressed streams of data from the server-side browser 104.The server-side browser 104 accesses information across the Internet or intranet 103.With respect to FIG.\",\n \"2, a user using a client device 206, 207 requests a Web page from a light weight browser or the light weight browser itself requests a Web page.\",\n \"The request is sent from the client 206, 207 to a server-side browser 201.The server-side browser 201 identifies the Web page that the user requested and sends a request to the proper server across the Internet or intranet.\",\n \"When the requested Web page is returned to the server-side browser 201, the server-side browser 201 loads the initial HTML document.\",\n \"The received HTML document consists of text, which comprises the content of the document, and tags, which define the structure and appearance of the document.\",\n \"The server-side browser 201 looks at any dependencies within the document (e.g., referenced images, Javascript files that need to be loaded remotely, etc.)\",\n \"and opens multiple sockets to get the required information across the Internet or intranet.\",\n \"The server-side browser 201 gathers the Web page information and lays out the information to calculate how to display the information on a particular device.\",\n \"The server-side browser 201 makes a second pass at the layout information and then transcodes the information into a very tight data stream that can be represented on the client 206, 207.The stream includes positional information that is not present in the normal HTML for the Web page.\",\n \"The server-side browser 201 takes all of the HTML information for a Web page and represents it as two dimensional content on the server.\",\n \"This two-dimensional representation helps the browser determine the exact position information positional information for all of the pieces of a particular HTML Web page.\",\n \"If there are any images or other dependencies, the server-side browser 201 scales (or crops) the images down and/or preprocesses the images (e.g., reduce the bit depth of an image, dither the image, etc.)\",\n \"to match the capabilities of the client 206, 207.The browser then creates a compressed stream that is sent down to the client.\",\n \"This compressed stream contains text and images with exact positional information (that tells where the pieces of content fit on the page), and hot spot information.\",\n \"The browser maps each rectangular hotspot region to a related URL.\",\n \"Thus, no extraneous information, such as hyperlink URLs, or Cookie Data is sent down to the client.\",\n \"The server-side browser 205 uses device characteristics information 202 for each supported device type, consisting of the associated default CSS style sheet, a list of supported fonts and their text metrics, device aspect ratios, pixel depth, color palette, form control dimensions, preferred layout algorithms, a DTD specifying level of HTML compliance, JavaScript policies, the User-Agent String to represent to a Web Server, and the compressed stream format encoder, to properly determine the ideal representation of Web content on the device.\",\n \"Various examples of potential clients with vastly different device characteristics could include: a cellphone 206 with a minimal CPU and minimal RAM; a PDA client with a small display but powerful CPU and high RAM; or a TV box 207 with a large display, high capacity CPU, low RAM, and non TCP/IP based communications.\",\n \"The server-side browser 205 utilizes many different layout algorithms to properly render information with these various classes of clients that all have different: display, CPU, RAM, and 10 constraints.\",\n \"A lightweight client browser is provided on the clients 206, 207 that accepts a standard information format from the server-side browser 201 and displays the data streams.\",\n \"This means that the server-side browser 201 can be updated at any time without affecting the clients 206, 207.Referring to FIGS.\",\n \"3, 4, and 5, a preferred embodiment of the invention provides a server-side browser engine 302 that accepts HTML (e.g., HTML 4.0 content with support for CSS version 1) pages 303 and parses the HTML 304.The parser 304 creates a Document Object Model (DOM) object hierarchy model 305.The DOM model 305 is a tree representation of the HTML in the Web page, which is well known in the art.\",\n \"A style sheet 306 tells the server-side browser 301 what fonts to use for standard HTML tags (e.g., bold), border sizes, and other stylistic information for each supported client device.\",\n \"A device information file 307 is also used by the browser that tells the server-side browser 301 the limitations of a client device (e.g., the size of a text control, fonts available, aspect ratio, etc.).\",\n \"The server-side browser 301 discovers what device type the client is when the client signs onto the server.\",\n \"Using the style sheet 306 and the device information 307 for the particular device, the server-side browser 301 creates a view tree 308 which is a two-dimensional model of the parent-child relationship between viewable objects of the DOM model 305.A DOM tree is shown in FIG.\",\n \"4.The HTML 401 is the root of the tree and underneath has a Body tag 402.Under the Body tag 402 is an Anchor tag 403 which has Text 404 underneath.\",\n \"There is also a Table 405 beneath the Anchor tag 403.The Table 405 has a row 406 which has a cell 407.The browser makes views of DOM tree elements that are visible.\",\n \"For example, a Text view 408 is made of the Text 404, a Table view 409 is made of the Table 405, a TR view 410 is made of the row 406, and a TD view 411 is made of the cell 407.An exemplary view model is shown in FIG.\",\n \"5, where a Body view 501 is the root of the tree.\",\n \"A Table view 502 sits below the Body view 501 and has a TR view 503 below it.\",\n \"A TD view 504 is below the TR view 503.The TR view 504 has both a text view 505 and an image view 506 below it.\",\n \"The server-side browser 301 then lays out the view model 308 on a virtual page.\",\n \"Each view in the view model 308 has a rectangular area of information attached to it.\",\n \"The server-side browser 301 lays out the views using specific layout algorithms for each view (e.g., tables, block levels, etc.)\",\n \"and uses the style sheet 306 and device information 307 for the particular device to make sure that any special characteristics or behaviors of the device are taken into account For example, for a TV box client, there may be a fixed table width defined in the view and fixed table widths are ignored by the TV box, or, one pixel width table borders are only supported on the TV box client.\",\n \"An example of a layout algorithm is every cell in a table is considered a row in itself, for a certain device each cell is to be laid out vertically instead of the normal horizontal layout.\",\n \"With respect to FIG.\",\n \"6, the server-side browser performs the first layout pass which lays out all of the views of the page 601.The page 601 is a virtual view of how the page looks.\",\n \"In this example, a body view 605 has a table view 604 within it.\",\n \"The Table view 604 has a row 603 which has a cell with text in it 602.Each view has a rectangular area that it represents and a parent-child relationship.\",\n \"This allows the server-side browser to walk through the viewable area to see what it represents.\",\n \"A second pass is made to determine what the real content is on the page.\",\n \"HTML authors often define multiply nested tables to constrain a layout of an HTML page.\",\n \"The server-side browser throws away the unnecessary information that does not contribute to the appearance of the page (e.g., not renderable).\",\n \"The server-side browser is concerned with the physical positioning of content elements within the page.\",\n \"For example, the browser finds text 602 on the page 601 and accumulates the text into a string and compresses it to send to the client.\",\n \"The server-side browser also sends the client information that tells the client where the text should be displayed in its viewing area.\",\n \"This information is placed in a binary format that the client device feeds into.\",\n \"The DOM tree and view tree and all state information (including Javascripts) for the client are preserved on the server.\",\n \"This is to ensure that, when the user is interacting with a page on the client, the server-side browser knows where the user has clicked on the page.\",\n \"The server-side browser does not send URL links to the client.\",\n \"This means that the client only has positional or hotspot information relating to where the user clicked or pressed on the page and sends that coordinate or hotspot information to the server-side browser.\",\n \"The server-side browser looks at the DOM tree and view model to find where the coordinates map onto the page.\",\n \"The view model is used to find the location of the coordinates or hotspot ID and the DOM tree is used to find the URL or script associated with the clicked area.\",\n \"When the server-side browser finds the area that corresponds to location where the user clicked, the server-side browser executes whatever script or URL request that corresponds with the area.\",\n \"The server-side browser then updates the client with the new page if the action was navigational, for example.\",\n \"Cookie data are impractical to place on the client device.\",\n \"The server-side browser caches all of the user's cookies in a user state that the server-side browser preserves on the server for each user.\",\n \"A session object exists on both the server and the client The session protocol is used by the client and server to communicate information.\",\n \"For example, when the client first signs onto the server, it tells the server its configuration information and other information.\",\n \"The server-side browser looks into its database and finds the client device's style and device information.\",\n \"Referring to FIG.\",\n \"7, a flowchart of an another preferred embodiment of the invention is shown.\",\n \"The client browser 711 sends a navigation request to the server-side browser engine 701.The browser session 702 retrieves the client specific style sheet 703 and layout algorithms 704.The browser session 702 fetches the requested Web page from the Internet or intranet.\",\n \"The HTML Web page 705 is parsed and a DOM tree model is created 706.The DOM tree model is then used to create a two-dimensional view tree model 707.The invention lays out the view tree based on the client specific style sheet and layout algorithms 708.Any additional layouts are performed on the view tree to fit the client device constraints 709.The resulting two-dimensional view tree and DOM tree are encoded in a lightweight format and sent to the client which decodes the format 710 and displays the resulting page to the user through the client browser 711.With respect to FIG.\",\n \"8, a task viewpoint of the invention is shown.\",\n \"The Client Browser module 812 determines the position of a user dick or press on its display screen and sends a navigation request containing the coordinates of the click or press, or the hotspot ID, to the server-side browser through the Client Session module 811 to the Server Session module 804.The Client Browser module 812 will also accept a specific URL from a user and places the URL in the navigation request.\",\n \"The Server Session module 804 forwards the navigation request to the Find Position module 805.The Find Position module 805 accepts Specific URLs from the client, coordinates of a user's click or press, and hotspot IDs 810.The Find Position module 805 forwards any specific URL to the Web Page Manager 902.Otherwise, the Find Position module 805 retrieves the DOM model and view tree from the storage device 807 for the particular client.\",\n \"It then finds the location on the Web page that the user clicked or pressed on using the coordinates or hotspot ID received from the client.\",\n \"If there is a script associated with the location, it is executed and the resulting page location is sent to the Web Page Manager 902.If there is a URL associated with the location, it is forwarded to the Web Page Manager 902.The Web Page Manager 802 receives the URL from the Find Position module 805 and formats a request for the Web page.\",\n \"The request is sent to the W e b Session module 803.The Web page is returned to the Web Session module 803 and is forwarded to the Web Page Manager 802.The Web page Manager 802 forwards the HTML Web page definition to the Parse HTML module 801.The Parse HTML module 801 parses the HTML and creates a DOM tree model which it stores on the storage device 807.The Web page Manager 802 then activates the Create View module 806.The Create View module 806 uses the DOM model stored on the storage device 807, creates a two-dimensional view tree model, and stores it on the storage device 807.Once the view tree is completed, the Web page Manager 802 retrieves the style sheet, layout algorithms, and device constraints for the specific client device are retrieved from the storage device 807.The Web page Manager 802 lays out the view model using the style sheet, layout algorithms, and device constraints.\",\n \"If the Client Browser 812 is a subset of the server-side browser engine, then the DOM tree and view tree are compressed into a stream 809 and sent through the Server Session module 804 to the Client Browser 812 via the Client Session module 811.The Client Browser 812 decodes the stream, translates the DOM tree and view tree, and displays the page to the user.\",\n \"Otherwise, the Web page Manager 802 performs the first layout pass and lays out all of the views of the page.\",\n \"The Web page Manager 802 then makes a second pass to determine the visual content on the page.\",\n \"Visual content and positional information are highly compressed and formatted into a stream 809 and forwarded to the Server Session module 804.The Server Session module 804 sends the stream to the Client Browser 812 via the Client Session module 811.The Client Browser 812 decodes the stream and displays the page to the user using the visual content and positional information.\",\n \"The DOM tree and view tree and all state information (including Javascripts and cookies) for the client are stored on the storage device 807 by the Web page Manager 802.Although the invention is described herein with reference to the preferred embodiment, one skilled in the art will readily appreciate that other applications may be substituted for those set forth herein without departing from the spirit and scope of the present invention.\",\n \"Accordingly, the invention should only be limited by the claims included below.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10474300"}}},{"rowIdx":797,"cells":{"text":{"kind":"list like","value":[["Thiazolyl-substsituted carbocylic 1,3-diones as pesticidal agents","The present invention relates to novel hetaryl-substituted carbocyclic 1,3-diones of the formula (I) in which A, B, Q1, Q2, Q3, Q4, G, m and Het are as defined in the description, to a plurality of processes for their preparation and to their use as pesticides, herbicides and fungicides."],["1.A compounds of the Formula (I) in which Het represents a nitrogen-containing 5-membered heterocycle, preferably selected from the group consisting of thiazolyl, which heterocycle is optionally substituted by halogen, alkyl, alkoxy, alkenyloxy, halogenoalkyl, halogenoalkoxy, halogenoalkenyloxy, cyano, nitro, alkylthio, alkylsulphinyl, alkylsulphonyl, optionally substituted phenyl or phenoxy, m represents the number 0 or 1, A represents hydrogen, represents in each case optionally halogen-substituted alkyl, alkenyl, alkoxyalkyl, polyalkoxyalkyl, alkylthioalkyl, saturated or unsaturated, optionally substituted cycloalkyl in which optionally at least one ring atom is replaced by a heteroatom, or in each case optionally halogen-, alkyl-, halogenoalkyl-, alkoxy-, halogenoalkoxy-, cyano- or nitro-substituted aryl, arylalkyl or hetaryl, B represents hydrogen or alkyl, A and B together with the carbon atom to which they are attached represent a saturated or unsaturated, unsubstituted or substituted cycle which optionally contains at least one heteroatom, A and Q1 together represent optionally substituted alkanediyl in which two not directly adjacent carbon atoms optionally form a further optionally substituted cycle, Q1 represents hydrogen, alkyl, alkoxyalkyl, optionally substituted cycloalkyl (in which optionally one methylene group is replaced by oxygen or sulphur) or in each case optionally substituted phenyl, hetaryl, phenylalkyl or hetarylalkyl, Q2, Q3, Q4 independently of one another represent hydrogen or alkyl, Q1 and Q2 together with the carbon atom to which they are attached represent a saturated or unsaturated, unsubstituted or substituted cycle which optionally contains one heteroatom, G represents hydrogen (a) or represents one of the groups in which E represents a metal ion or an ammonium ion, L represents oxygen or sulphur, M represents oxygen or sulphur, R1 represents in each case optionally halogen-substituted alkyl, alkenyl, alkoxyalkyl, alkylthioalkyl, polyalkoxyalkyl or optionally halogen-, alkyl- or alkoxy-substituted cycloalkyl which may be interrupted by at least one heteroatom, in each case optionally substituted phenyl, phenylalkyl, hetaryl, phenoxyalkyl or hetaryloxyalkyl, R2 represents in each case optionally halogen-substituted alkyl, alkenyl, alkoxyalkyl, polyalkoxyalkyl or represents in each case optionally substituted cycloalkyl, phenyl or benzyl, R3 represents in each case optionally substituted alkyl, halogenoalkyl, phenyl or benzyl, R4 and R5 independently of one another represent in each case optionally halogen-substituted alkyl, alkoxy, alkylamino, dialkylamino, alkylthio, alkenylthio, cycloalkylthio or represent in each case optionally substituted phenyl, benzyl, phenoxy or phenylthio, R6 and R7 independently of one another represent hydrogen, in each case optionally halogen-substituted alkyl, cycloalkyl, alkenyl, alkoxy, alkoxyalkyl, represent optionally substituted phenyl, represent optionally substituted benzyl, or together with the N atom to which they are attached represent a cycle which is optionally interrupted by oxygen or sulphur.","2.A compounds of the formula (I) according to claim 1, in which Het represents m represents the number 0 or 1, X represents hydrogen, halogen, C1-C6-alkyl, C1-C6-alkoxy, C3-C6-alkenyloxy, nitro or cyano, Y represents halogen, C1-C6-alkyl, C1-C6-halogenoalkyl, C1-C6-alkoxy, C1-C6-halogenoalkoxy or represents the groups V1 represents hydrogen, halogen, C1-C12-alkyl, C1-C6-alkoxy, C1-C6-alkylthio, C1-C6-alkylsulphinyl, C1-C6-alkylsulphonyl, C1-C4-halogenoalkyl, C1-C4-halogenoalkoxy, nitro, cyano or represents phenyl, phenoxy, phenoxy-C1-C4-alkyl, phenyl-C1-C4-alkoxy, phenylthio-C1-C4-alkyl or phenyl-C1-C4-alkylthio, each of which is optionally mono- or polysubstituted by halogen, C1-C6-alkyl, C1-C6-alkoxy, C1-C4-halogenoalkyl, C1-C4-halogenoalkoxy, nitro or cyano, V2 and V3 independently of one another represent hydrogen, halogen, C1-C6-alkyl, C1-C6-alkoxy, C1-C4-halogenoalkyl or C1-C4-halogeno-alkoxy, V1 and V2 jointly together with the carbon atoms to which they are attached represent an optionally C1-C4-alkyl- or halogen-substituted 5- or 6-membered cycle in which optionally one or two carbon atoms may be replaced by oxygen, sulphur or nitrogen, A represents hydrogen or in each case optionally halogen-substituted C1-C12-alkyl, C3-C8-alkenyl, C1-C6-alkoxy-C1-C4-alkyl, optionally halogen-, C1-C4-alkyl- or C1-C4-alkoxy-substituted C3-C8-cycloalkyl or C3-C6-cycloalkyl-C1-C4-alkyl, in which optionally one or two not directly adjacent ring members are replaced by oxygen and/or sulphur, or represents in each case optionally halogen-, C1-C6-alkyl-, C1-C6-halogenoalkyl-, C1-C6-alkoxy-, C1-C6-halogenoalkoxy-, cyano- or nitro-substituted phenyl, benzyl, hetaryl having 5 or 6 ring atoms or hetaryl-C1-C4-alkyl having 5 or 6 ring atoms, B represents hydrogen or C1-C6-alkyl, A, B and the carbon atom to which they are attached represent saturated C3-C10-cycloalkyl or unsaturated C5-C10-cycloalkyl in which optionally one ring member is replaced by oxygen or sulphur and which are optionally mono- or disubstituted by C1-C6-alkyl, C3-C8-cycloalkyl, C1-C6-halogenoalkyl, C1-C6-alkoxy, C1-C6-alkylthio, halogen or phenyl, A and Q1 together represent C3-C6-alkanediyl which is optionally mono- or disubstituted by identical or different substituents from the group consisting of C1-C4-alkyl and C1-C4-alkoxy, Q1 represents hydrogen, C1-C6-alkyl, C1-C6-alkoxy-C1-C2-alkyl, optionally fluorine-, chlorine-, C1-C4-alkyl-, C1-C2-halogenoalkyl- or C1-C4-alkoxy-substituted C3-C8-cycloalkyl in which optionally one methylene group is replaced by oxygen or sulphur or in each case optionally halogen-, C1-C4-alkyl-, C1-C4-alkoxy-, C1-C2-halogenoalkyl-, C1-C2-halogenoalkoxy-, cyano- or nitro-substituted phenyl, pyridyl, thienyl, thiazolyl, phenyl-C1-C4-alkyl, pyridyl-C1-C2-alkyl or thiazolyl-C1-C2-alkyl, Q2, Q3, Q4 independently of one another represent hydrogen or C1-C4-alkyl, Q1 and Q2 together with the carbon atom to which they are attached represent optionally C1-C6-alkyl-, C1-C6-alkoxy- or C1-C2-halogenoalkyl-substituted C3-C7-cycloalkyl in which optionally one ring member is replaced by oxygen or sulphur, G represents hydrogen (a) or represents one of the groups in which E represents a metal ion or an ammonium ion, L represents oxygen or sulphur and M represents oxygen or sulphur, R1 represents in each case optionally halogen-substituted C1-C20-alkyl, C2-C20-alkenyl, C1-C8-alkoxy-C1-C8-alkyl, C1-C8-alkylthio-C1-C8-alkyl, poly-C1-C8-alkoxy-C1-C8-alkyl or optinally halogen-, C1-C6-alkyl- or C1-C6-alkoxy-substituted C3-C8-cycloalkyl, in which optionally one or more not directly adjacent ring members are replaced by oxygen and/or sulphur, represents optionally halogen-, cyano-, nitro-, C1-C6-alkyl, C1-C6-alkoxy-, C1-C6-halogenoalkyl-, C1-C6-halogenoalkoxy-, C1-C6-alkylthio or C1-C6-alkylsulphonyl-substituted phenyl, represents optionally halogen-, nitro-, cyano-, C1-C6-alkyl-, C1-C6-alkoxy-, C1-C6-halogenoalkyl- or C1-C6-halogenoalkoxy-substituted phenyl-C1-C6-alkyl, represents optionally halogen-, C1-C6-alkyl- or trifluoromethyl-substituted 5- or 6-membered hetaryl, represents optionally halogen- or C1-C6-alkyl-substituted phenoxy-C1-C6-alkyl or represents optionally halogen-, amino- or C1-C6-alkyl-substituted 5- or 6-membered hetaryloxy-C1-C6-alkyl, R2 represents in each case optionally halogen-substituted C1-C20-alkyl, C2-C20-alkenyl, C1-C8-alkoxy-C2-C8-alkyl, poly-C1-C8-alkoxy-C2-C8-alkyl, represents optionally halogen-, C1-C6-alkyl- or C1-C6-alkoxy-substituted C3-C8-cycloalkyl or represents in each case optionally halogen-, cyano-, nitro-, C1-C6-alkyl-, C1-C6-alkoxy-, C1-C6-halogenoalkyl- or C1-C6-halogenoalkoxy-substituted phenyl or benzyl, R3 represents optionally halogeno-substituted C1-C8-alkyl or represents in each case optionally halogen-, C1-C6-alkyl-, C1-C6-alkoxy-, C1-C4-halogenoalkyl-, C1-C4-halogenoalkoxy-, cyano- or nitro-substituted phenyl or benzyl, R4 and R5 independently of one another represent in each case optionally halogen-substituted C1-C8-alkyl, C1-C8-alkoxy, C1-C8-alkylamino, di-(C1-C8-alkyl)amino, C1-C8-alkylthio, C2-C8-alkenylthio, C3-C7-cycloalkylthio or represent in each case optionally halogen-, nitro-, cyano-, C1-C4-alkoxy-, C1-C4-halogenoalkoxy-, C1-C4-alkylthio-, C1-C4-halogenoalkylthio-, C1-C4-alkyl- or C1-C4-halogenoalkyl-substituted phenyl, benzyl, phenoxy or phenylthio, R6 and R7 independently of one another represent hydrogen, represent in each case optionally halogen-substituted C1-C8-alkyl, C3-C8-cycloalkyl, C1-C8-alkoxy, C3-C8-alkenyl, C1-C8-alkoxy-C1-C8-alkyl, represent optionally halogen-, C1-C8-halogenoalkyl-, C1-C8-alkyl- or C1-C8-alkoxy-substituted phenyl, optionally halogen-, C1-C8-alkyl-, C1-C8-halogenoalkyl- or C1-C8-alkoxy-substituted benzyl or together with the nitrogen to which they are attached represent an optionally C1-C4-alkyl-substituted C3-C6-alkylene radical in which optionally one carbon atom is replaced by oxygen or sulphur.","3.A compound of the Formula (I) according to claim 1, in which Het represents m represents the number 0 or 1, X represents hydrogen, chlorine, bromine or C1-C4-alkyl, Y represents chlorine, bromine, C1-C4-alkyl, C1-C4-halogenoalkyl, C1-C4-alkoxy, C1-C4-halogenoalkoxy or the group V1 represents hydrogen, fluorine, chlorine, C1-C6-alkyl, C1-C4-alkoxy, C1-C2-halogenoalkyl, C1-C2-halogenoalkoxy, nitro, cyano or phenoxy which is optionally mono- or disubstituted by fluorine, chlorine, bromine, C1-C4-alkyl, C1-C4-alkoxy, C1-C2-halogenoalkyl, C1-C2-halogenoalkoxy, nitro or cyano, V2 represents hydrogen, fluorine, chlorine, bromine, C1-C4-alkyl, C1-C4-alkoxy, C1-C2-halogenoalkyl or C1-C2-halogenoalkoxy, V1 and V2 jointly, together with the carbon atoms to which they are attached, represent an optionally fluorine- or methyl-substituted 5- or 6-membered cycle in which optionally one or two carbon atoms may be replaced by oxygen, A represents hydrogen, represents C1-C8-alkyl or C1-C4-alkoxy-C1-C2-alkyl, each of which is optionally mono- to trisubstituted by fluorine, represents C5-C6-cycloalkyl or C3-C6-cycloalkyl-C1-C2-alkyl in which optionally one ring member is replaced by oxygen or sulphur, each of which radicals is optionally mono- or disubstituted by fluorine, chlorine, methyl, ethyl or methoxy, or represents phenyl or benzyl, each of which is optionally mono- or disubstituted by fluorine, chlorine, bromine, C1-C4-alkyl, C1-C2-halogenoalkyl, C1-C4-alkoxy or C1-C2-halogenoalkoxy, B represents hydrogen or C1-C4-alkyl, A, B and the carbon atom to which they are attached represent saturated C5-C6-cycloalkyl in which optionally one ring member is replaced by oxygen and which is optionally monosubstituted by C1-C4-alkyl, trifluoromethyl or C1-C4-alkoxy, with the proviso that in this case Q1 represents only hydrogen or C1-C4-alkyl, A and Q1 together represent C3-C4-alkanediyl which is optionally mono- or disubstituted by methyl, ethyl, methoxy or ethoxy, Q1 represents hydrogen, C1-C4-alkyl, C1-C4-alkoxy-C1-C2-alkyl, or optionally methyl- or methoxy-substituted C3-C6-cycloalkyl, in which optionally one methylene group is replaced by oxygen, or represents phenyl or benzyl, each of which is optionally mono- or disubstituted by fluorine, chlorine, bromine, C1-C4-alkyl, C1-C4-alkoxy, trifluoromethyl or trifluoromethoxy, Q2, Q3, Q4 independently of one another represent hydrogen, methyl or ethyl, Q1 and Q2 together with the carbon atom to which they are attached represent optionally C1-C4-alkyl- or C1-C4-alkoxy-substituted saturated C5-C6-cycloalkyl in which optionally one ring member is replaced by oxygen, with the proviso that in this case A only represents hydrogen or C1-C4-alkyl, G represents hydrogen (a) or represents one of the groups in which E represents a metal ion or an ammonium ion, L represents oxygen or sulphur and M represents oxygen or sulphur, R1 represents C1-C16-alkyl, C2-C16-alkenyl, C1-C4-alkoxy-C1-C2-alkyl, C1-C4-alkylthio-C1-C2-alkyl, each of which is optionally mono- to pentasubstituted by fluorine or chlorine, or C3-C6-cycloalkyl in which optionally one or two not directly adjacent ring members are replaced by oxygen and/or sulphur and which is optionally mono- or disubstituted by fluorine, chlorine, C1-C4-alkyl or C1-C5-alkoxy, represents phenyl which is optionally mono- or disubstituted by fluorine, chlorine, bromine, cyano, nitro, C1-C4-alkyl, C1-C4-alkoxy, trifluoromethyl or trifluoromethoxy, represents pyridyl or thienyl, each of which is optionally monosubstituted by fluorine, chlorine, bromine, methyl, ethyl or trifluoromethyl, R2 represents C1-C16-alkyl, C2-C16-alkenyl or C 1 -C4-alkoxy-C2-C4-alkyl, represents C3-C6-cycloalkyl which is optionally mono- or disubstituted by methyl, ethyl or methoxy, represents phenyl or benzyl, each of which is optionally mono- or disubstituted by fluorine, chlorine, bromine, cyano, nitro, C1-C4-alkyl, C1-C4-alkoxy, trifluoromethyl or trifluoromethoxy, R3 represents C1-C4-alkyl which is optionally mono- to pentasubstituted by fluorine or represents phenyl which is optionally mono- or disubstituted by fluorine, chlorine, bromine, C1-C4-alkyl, C1-C4-alkoxy, trifluoromethyl, trifluoromethoxy, cyano or nitro, R4 represents C1-C6-alkyl, C1-C6-alkoxy, C1-C6-alkylamino, di-(C1-C6-alkyl)amino, C1-C6-alkylthio, or represents phenyl, benzyl, phenoxy or phenylthio, each of which is optionally mono- or disubstituted by fluorine, chlorine, bromine, nitro, cyano, C1-C3-alkoxy, trifluoromethoxy, C1-C3-alkyl or trifluoromethyl, R5 represents C1-C4-alkyl, C1-C4-alkoxy or C1-C4-alkylthio, R6 represents hydrogen, C1-C6-alkyl, C3-C6-cycloalkyl, C1-C6-alkoxy, C3-C6-alkenyl, C1-C6-alkoxy-C1-C6-alkyl, represents phenyl which is optionally mono- or disubstituted by fluorine, chlorine, bromine, trifluoromethyl, C1-C4-alkyl or C1-C4-alkoxy, represents benzyl which is optionally substituted by fluorine, chlorine, bromine, methyl, ethyl, trifluoromethyl or methoxy, R7 represents hydrogen, C1-C6-alkyl or C3-C6-alkenyl, R6 and R7 together with nitrogen atom to which they are attached represent a C5-C6-alkylene radical in which optionally one methylene group is replaced by oxygen or sulphur and which is optionally mono- or disubstituted by methyl or ethyl.","4.A compound of the Formula (I) according to claim 1, in which Het represents m represents the number 0 or 1, X represents hydrogen, chlorine, bromine, methyl, ethyl, n-propyl or isopropyl, Y represents the group V1 represents hydrogen, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, methoxy, ethoxy, n-propoxy, isopropoxy, trifluoromethyl, trifluoromethoxy or 4-chlorophenoxy, V2 represents hydrogen, fluorine, chlorine, bromine, methyl, methoxy or trifluoromethyl, A represents hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, methoxymethyl, ethoxymethyl, phenyl or cyclohexyl, B represents hydrogen, methyl or ethyl, A, B and the carbon atom to which they are attached represent saturated C5-C6-cycloalkyl in which optionally one ring member is replaced by oxygen and which is optionally monosubstituted by methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, trifluoromethyl, methoxy, ethoxy, n-propoxy, n-butoxy or isobutoxy, with the provisio that in this case Q1 only represents hydrogen, A and Q1 together represent C3-C4-alkanediyl, Q1 represents hydrogen, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, cyclopentyl, cyclohexyl or 4-chlorophenyl, Q2, Q3, Q4 independently of one another represent hydrogen, methyl or ethyl, Q1 and Q2 together with the carbon atom to which they are attached represent optionally methyl-, ethyl-, methoxy-, ethoxy-, n-propoxy- or n-butoxy-substituted saturated C5-C6-cycloalkyl in which optionalloy one ring member is replaced by oxygen, with the proviso that in this case A only represents hydrogen, G represents hydrogen (a) or represents one of the groups in which E represents a metal ion or an ammonium ion, L represents oxygen or sulphur and M represents oxygen or sulphur, R1 represents in each case optionally fluorine- or chlorine-substituted C1-C8-alkyl, C2-C8-alkenyl, C1-C2-alkoxy-C1-C2-alkyl, C1-C2-alkylthio-C1-C2-alkyl or cyclopropyl or cyclohexyl, each of which is optionally monosubstituted by fluorine, chlorine, methyl, ethyl or methoxy, represents phenyl which is optionally monosubstituted by fluorine, chlorine, bromine, cyano, nitro, methyl, ethyl, isopropyl, tert-butyl, methoxy, trifluoromethyl or trifluoromethoxy, represents thienyl or pyridyl, each of which is optionally monosubstituted by fluorine, chlorine, bromine or methyl, R2 represents C1-C8-alkyl, C2-C8-alkenyl or C1-C4-alkoxy-C2-alkyl, represents C3-C6-cycloalkyl which is optionally monosubstituted by methyl, ethyl or methoxy, represents phenyl or benzyl, each of which is optionally monosubstituted by fluorine, chlorine, cyano, nitro, methyl, ethyl, isopropyl, tert-butyl, methoxy, trifluoromethyl or trifluoromethoxy, R3 represents methyl or ethyl, each of which is optionally trisubstituted by fluorine or represents phenyl which is optionally monosubstituted by fluorine, chlorine, bromine, methyl, tert-butyl, methoxy, trifluoromethyl, trifluoromethoxy, cyano or nitro, R4 represents C1-C4-alkyl, C1-C4-alkoxy, C1-C4-alkylamino, di-(C1-C4-alkyl)amino, C1-C4-alkylthio or represents phenyl, phenoxy or phenylthio, each of which is optionally monosubstituted by fluorine, chlorine, bromine, nitro, cyano, C1-C2-alkoxy, trifluoromethoxy or C1-C3-alkyl, R5 represents methyl, ethyl, methoxy, ethoxy, methylthio or ethylthio, R6 represents hydrogen, represents C1-C4-alkyl, C3-C6-cycloalkyl, C1-C4-alkoxy, C3-C4-alkenyl or C1-C4-alkoxy-C1-C4-alkyl, R7 represents hydrogen, C1-C4-alkyl or C3-C4-alkenyl, R6 and R7 together with the nitrogen atom to which they are attached represent a C5-alkylene radical in which optionally one methylene group is replaced by oxygen.","5.A compound of the Formula (I) according to claim 1, in which Het represents m represents the number 0 or 1, X represents hydrogen, methyl or ethyl, Y represents the group V1 represents hydrogen, chlorine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, trifluoromethyl or represents 4-chloro-phenoxy, A represents hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, phenyl or cyclohexyl, B represents hydrogen, methyl or ethyl, A, B and the carbon atom to which they are attached represent saturated C5-C6-cycloalkyl in which optionally one ring member is replaced by oxygen and which is optionally monosubstituted by methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, methoxy, ethoxy, n-propoxy, n-butoxy or isobutoxy, with the proviso that in this case Q1 only represents hydrogen, A and Q1 together represent C3-C4-alkanediyl, Q1 represents hydrogen, methyl, ethyl, n-propyl, isopropyl or 4-chlorophenyl, Q2 represents hydrogen, methyl or ethyl, Q3 represents hydrogen, Q4 represents hydrogen, Q1 and Q2 together with the carbon atom to which they are attached represent saturated C5-C6-cycloalkyl, with the proviso that in this case A only represents hydrogen, G represents hydrogen (a) or represents one of the groups in which L represents oxygen and M represents oxygen or sulphur, R1 represents C1-C8-alkyl, C1-C2-alkoxy-C1-C2-alkyl or cyclopropyl which is optionally monosubstituted by chlorine, represents phenyl which is optionally monosubstituted by chlorine, represents pyridyl which is optionally monosubstituted by chlorine, R2 represents C1-C8-alkyl, represents phenyl or benzyl, R3 represents methyl or ethyl, R6 and R7 together with the nitrogen atom to which they are attached represent a C5-alkylene radical in which optionally one methylene group is replaced by oxygen.","6.A process for preparing a compound of the Formula (I) according to claim 1, wherein to obtain (A) a compound of the Formula (I-1-a) in which A, B, Q1, Q2 and Het are as defined in claim 1, a compound of the Formula (II) in which A, B, Q1, Q2 and Het are as defined in claim 1 and R8 represents alkyl, is condensed intramolecularly, optionally in the presence of a diluent and in the presence of a base, (B) a compound of the Formula (I-2-a) in which A, B, Q1, Q2, Q3, Q4, and Het are as defined claim 1, a compound of the Formula (III) in which A, B, Q1, Q2, Q3, Q4 and Het are as defined in claim 1 and R8 represents alkyl, is condensed intramolecularly in the presence of a diluent and in the presence of a base, (C) a compound of the Formulae (I-1-b) or (I-2-b) in which A, B, Q1, Q2, Q3, Q4, m, R1 and Het are as defined in claim 1, a compound of the Formulae (I-1-a) or (I-2-a) shown above in this claim 6 in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined in claim 1 are in each case (α) reacted with an acid halide of the Formula (IV) in which R1 is as defined in claim 1 and Hal represents halogen or (β) reacted with a carboxylic anhydride of the Formula (V) R1—CO—O—CO—R1 (V) in which R1 is as defined in claim 1, optionally in the presence of a diluent and optionally in the presence of an acid binder; (D) a compound of the Formulae (I-1-c) or (I-2-c) in which A, B, Q1 , Q2, Q3, Q4, m, R2, M and Het are as defined in claim 1 and L represents oxygen, compounds of the Formulae (I-1-a) or (I-2-a) shown above in this claim 6 in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined in claim 1 are in each case reacted with a chloroformic ester or a chloroformic thioester of the Formula (VI) R2—M—CO—Cl (VI) in which R2 and M are as defined in claim 1, optionally in the presence of a diluent and optionally in the presence of an acid binder; (E) a compound of the Formulae (I-1-c) or (I-2-c) shown above in this claim 6 in which A, B, Q1, Q2, Q3, Q4, m, R2, M and Het are as defined in claim 1 and L represents sulphur, compounds of the Formulae (I-1 -a) or (I-2-a) shown above in this claim 6 in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined in claim 1 are in each case reacted with a chloromonothioformic ester or a chlorodithioformic ester of the Formula (VII) in which M and R2 are as defined in claim 1, optionally in the presence of a diluent and optionally in the presence of an acid binder and (F) a compound of the Formulae (I-1-d) or (I-2-d) in which A, B, Q1, Q2, Q3, Q4, m, R3 and Het are as defined in claim 1, compounds of the Formulae (I-1-a) or (I-2-a) shown above in this claim 6 which A, B, Q1, Q2, Q3, Q4, m and Het are as defined in claim 1 are in each case reacted with a sulphonyl chloride of the Formula (VIII) R3—SO2—Cl (VIII) in which R3 is as defined in claim 1, optionally in the presence of a diluent and optionally in the presence of an acid binder, (G) a compound of the Formulae (I-1-e) or (I-2-e) in which A, B, L, Q1, Q2, Q3, Q4, m, R4, R5 and Het are as defined in claim 1, compounds of the Formulae (I-1-a) or (I-2-a) shown above in this claim 6 in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined in claim 1 are in each case reacted with a phosphorus compound of the Formula (IX) in which L, R4 and R5 are as defined in claim 1 and Hal represents halogen, optionally in the presence of a diluent and optionally in the presence of an acid binder, (H) a compound of the formulae (I-1-f) or (I-2-f) in which A, B, E, Q1, Q2, Q3, Q4, m and Het are as defined in claim 1, compounds of the Formulae (I-1-a) or (I-2-a) shown above in this claim 6 in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined in claim 1 are in each case reacted with a metal compound or an amine of the Formulae (X) and (XI), respectively, Me(OR11)t (X) in which Me represents a mono- or divalent metal, t represents the number 1 or 2 and R11, R12, R13 independently of one another represent hydrogen or alkyl, optionally in the presence of a diluent, (I) a compound of the Formulae (I-1-g) or (I-2-g) in which A, B, L, Q1, Q2, Q3, Q4, m, R6, R7 and Het are as defined in claim 1, compounds of the Formulae (I-1-a) or (I-2-a) shown above in this claim 6 in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined in claim 1 are in each case (α) reacted with an isocyanater or isothiocyanater of the Formula (XII) R6—N═C═L (XII) in which R6 and L are as defined in claim 1, optionally in the presence of a diluent and optionally in the presence of a catalyst, or (β) reacted with a carbamoyl chloride or a thiocarbamoyl chloride of the Formula (XIII) in which L, R6 and R7 are as defined in claim 1, optionally in the presence of a diluent and optionally in the presence of an acid binder.","7.A compound of the Formula (II) in which A, B, Q1, Q2 and Het are as defined in claim 1 and R8 represents alkyl.","8.A compound of the Formula (III) in which A, B, Q1, Q2, Q3, Q4, and Het are as defined in claim 1, and R8 represents alkyl.","9.A compound of the Formula (XIV) in which A, B, Het, Q1 and Q2 are as defined in claim 1.10.A compound of the Formula (XV) in which A, B, Het, Q1 and Q2 are as defined claim 1 and R8 and R8′ represents alkyl.","11.A compound of the Formula (XIX) in which A, B, Het, Q1, Q2, Q3 and Q4 are as defined claim 1.12.A compound of the Formula (XX) in which A, B, Het, Q1, Q2, Q3, and Q4are as defined in claim 1, R8 and R8 represent alkyl.","13.Pesticides, herbicides and fungicides comprising at least one compound of the Formula (I) according to claim 1.14.A method for controlling animal pests, undesirable vegetation and/or fungi comprising allowing a compound of the Formula (I) according to claim 1 to act on pests, undesirable vegetation and/or fungi and/or their habitat.","15.canceled 16.A process for preparing pesticides, herbicides and/or fungicides comprising mixing one or more compounds of the Formula (I) according to claim 1 with one or more extenders and/or surfactants.","17.canceled."],["The present invention relates to novel hetaryl-substituted cyclopentane- and cyclohexane-1,3-dione derivatives, to a plurality of processes for their preparation and to their use as bioactive compounds in agriculture.","2-(2-Amino-thiazol-4-yl)-3-hydroxy-5,5-dimethyl-cyclohex-2-enone is known; Pshenichnyi, V. N. et al; CNClALi; Chem.","Heterocycl.","Compd.","(Engl.","transl.)","EN; 26, 1175-1178 (1990).","This invention now provides novel compounds of the formula (I) in which Het represents a nitrogen-containing 5-membered heterocycle, preferably from the group consisting of thiazolyl, which heterocycle is optionally substituted by halogen, alkyl, alkoxy, alkenyloxy, halogenoalkyl, halogenoalkoxy, halogenoalkenyloxy, cyano, nitro, alkylthio, alkylsulphinyl, alkylsulphonyl, optionally substituted phenyl or phenoxy, m represents the number 0 or 1, A represents hydrogen, represents in each case optionally halogen-substituted alkyl, alkenyl, alkoxyalkyl, polyalkoxyalkyl, alkylthioalkyl, saturated or unsaturated, optionally substituted cycloalkyl in which optionally at least one ring atom is replaced by a heteroatom, or in each case optionally halogen-, alkyl-, halogenoalkyl-, alkoxy-, halogenoalkoxy-, cyano- or nitro-substituted aryl, arylalkyl or hetaryl, B represents hydrogen or alkyl, A and B together with the carbon atom to which they are attached represent a saturated or unsaturated, unsubstituted or substituted cycle which optionally contains at least one heteroatom, A and Q1 together represent optionally substituted alkanediyl in which two not directly adjacent carbon atoms optionally form a further optionally substituted cycle, Q1 represents hydrogen, alkyl, alkoxyalkyl, optionally substituted cycloalkyl (in which optionally one methylene group is replaced by oxygen or sulphur) or in each case optionally substituted phenyl, hetaryl, phenylalkyl or hetarylalkyl, Q2, Q3, Q4 independently of one another represent hydrogen or alkyl, Q1 and Q2 together with the carbon atom to which they are attached represent a saturated or unsaturated, unsubstituted or substituted cycle which optionally contains one heteroatom, G represents hydrogen (a) or represents one of the groups in which E represents a metal ion or an ammonium ion, L represents oxygen or sulphur, M represents oxygen or sulphur, R1 represents in each case optionally halogen-substituted alkyl, alkenyl, alkoxyalkyl, alkylthioalkyl, polyalkoxyalkyl or optionally halogen-, alkyl- or alkoxy-substituted cycloalkyl which may be interrupted by at least one heteroatom, in each case optionally substituted phenyl, phenylalkyl, hetaryl, phenoxyalkyl or hetaryloxyalkyl, R2 represents in each case optionally halogen-substituted alkyl, alkenyl, alkoxyalkyl, polyalkoxyalkyl or represents in each case optionally substituted cycloalkyl, phenyl or benzyl, R3 represents in each case optionally substituted alkyl, halogenoalkyl, phenyl or benzyl, R4 and R5 independently of one another represent in each case optionally halogen-substituted alkyl, alkoxy, alkylamino, dialkylamino, alkylthio, alkenylthio, cycloalkylthio or represent in each case optionally substituted phenyl, benzyl, phenoxy or phenylthio, R6 and R7 independently of one another represent hydrogen, in each case optionally halogen-substituted alkyl, cycloalkyl, alkenyl, alkoxy, alkoxyalkyl, represent optionally substituted phenyl, represent optionally substituted benzyl, or together with the N atom to which they are attached represent a cycle which is optionally interrupted by oxygen or sulphur.","Depending inter alia on the nature of the substituents, the compounds of the formula (I) can be present as geometrical and/or optical isomers or isomer mixtures of varying composition which, if desired, can be separated in a known manner.","The present invention provides both the pure isomers and the isomer mixtures, their preparation and use and the compositions comprising them.","However, hereinbelow, for the sake of simplicity, only compounds of the formula (I) are referred to, although this includes both the pure compounds and, if appropriate, mixtures having varying proportions of isomeric compounds.","Depending on the position of the substituent G, the compounds of the formula (I) can be present in the two isomeric forms of the formulae (I-α) and (I-β), which is meant to be expressed by the broken line in the formula (I).","The compounds of the formulae (I-α) and (I-β) can be present either as mixtures or else in the form of their pure isomers.","Mixtures of the compounds of the formulae (I-α) and (I-β) can, if desired, be separated in a manner known per se by physical methods, for example by chromatographic methods.","For reasons of clarity, in each case only one of the possible isomers is shown hereinbelow.","This does not preclude that the compounds are, if appropriate, present in the form of the isomer mixtures or in the respective other isomeric form.","Including Het for thiazolyl and m for the numbers 0 and 1, the following principal structures (I-1) and (I-2) result Including the different meanings (a), (b), (c), (d,(e), (f) and (g) of the group G. the following principal structures (I-1-a) to (I-1-g) result if m represents the number 0 (I-1), in which A, B, E, L, M, Q1, Q2, R1, R2, R3, R4, R5, R6 and R7 are as defined above, and x represents hydrogen, halogen, alkyl, alkoxy, alkenyloxy, nitro, cyano or optionally substituted phenyl and Y represents halogen, alky, halogenoalkyl, alkoxy, halogenoalkoxy or in each case optionally substituted phenyl or phenoxy.","Including the different meanings (a), (b), (c), (d), (e), (f) and (g) of the group G, the following principal structures (I-2-a) to (I-2-g) result if m represents the number 1 (I-2), in which A, B, E, L, M, Q1, Q2, Q3, Q4, X, Y, R1, R2, R3, R4, R5, R6 and R7 are as defined above.","Furthermore, it has been found that the novel compounds of the formula (I) are obtained by one of the processes described below: (A) substituted cyclopentane-1,3-diones or their enoles of the formula (I-1-a) in which A, B, Q1, Q2 and Het are as defined above, are obtained when ketocarboxylic esters of the formula (II) in which A, B, Q1, Q2 and Het are as defined above and R8 represents alkyl (preferably C1-C6-alkyl) are condensed intramolecularly, if appropriate in the presence of a diluent and in the presence of a base.","Furthermore, it has been found (B) that substituted cyclohexane-1,3-diones of the formula (1-2-a) in which A, B, Q1, Q2, Q3, Q4, and Het are as defined above are obtained when ketocarboxylic esters of the formula (III) in which A, B. Q, Q2, Q3, Q4, and Het are as defined above, and R8 represents alkyl (preferably C1-C6-alkyl) are condensed intramolecularly in the presence of a diluent and in the presence of a base.","Furthermnore, it has been found (C) that the compounds of the formulae (I-1-b) to (I-2-b) shown above in which A, B, Q1, Q2, Q3, Q4, m, R1 and Het are as defined above are obtained when compounds of the formulae (I-1-a) to (I-2-a) shown above in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined above are in each case (α) reacted with acid halides of the formula (IV) in which R1 is as defined above and Hal represents halogen (in particular chlorine or bromine) or (β) reacted with carboxylic anhydrides of the formula (V) R1—CO—O—CO—R1 (V) in which R1 is as defined above, if appropriate in the presence of a diluent and if appropriate in the presence of an acid binder; (D) that the compounds of the formulae (I-1-c) to (I-2-c) shown above in which A, B, Q1, Q2, Q3, Q4, m, R2, M and Het are as defined above and L represents oxygen are obtained when compounds of the formulae (I-1-a) to (I-2-a) shown above in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined above are in each case reacted with chloroformic esters or chloroformic thioesters of the formula (VI) R2—M—CO—Cl (VI) in which R2 and M are as defined above, if appropriate in the presence of a diluent and if appropriate in the presence of an acid binder; (E) that compounds of the formulae (I-1-c) to (I-2-c) shown above in which A, B, Q1, Q2, Q3, Q4, m, R2, M and Het are as defined above and L represents sulphur are obtained when compounds of the formulae (I-1-a) to (I-2-a) shown above in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined above are in each case reacted with chloromonothioformic esters or chlorodithioformic esters of the formula (VII) in which M and R2 are as defined above, if appropriate in the presence of a diluent and if appropriate in the presence of an acid binder and (F) that compounds of the formulae (I-1-d) to (I-2-d) shown above in which A, B, Q1, Q2, Q3, Q4, m, R3 and Het are as defined above are obtained when compounds of the formulae (I-1-a) and (I-2-a) shown above in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined above are in each case reacted with sulphonyl chlorides of the formula (VIII) R3—SO2—Cl (VIII) in which R3 is as defined above, if appropriate in the presence of a diluent and if appropriate in the presence of an acid binder, (G) that compounds of the formulae (I-1-e) to (I-2-e) shown above in which A, B, L, Q1, Q2, Q3, Q4, m, R4, R5 and Het are as defined above are obtained when compounds of the formulae (I-1-a) to (I-2-a) shown above in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined above are in each case reacted with phosphorus compounds of the formula (IX) in which L, R4 and R5 are as defined above and Hal represents halogen (in particular chlorine or bromine), if appropriate in the presence of a diluent and if appropriate in the presence of an acid binder, (H) that compounds of the formulae (I-1-f) to (I-2-f) shown above in which A, B, E, Q1, Q2, Q3, Q4, m and Het are as defined above are obtained when compounds of the formulae (I-1-a) to (I-2-a) in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined above are in each case reacted with metal compounds or amines of the formulae (X) and (XI), respectively, in which Me represents a mono- or divalent metal (preferably an alkali metal or alkaline earth metal such as lithium, sodium, potassium, magnesium or calcium), t represents the number 1 or 2 and R11, R12, R13 independently of one another represent hydrogen or alkyl (preferably C1-C8-alkyl), if appropriate in the presence of a diluent, (I) that compounds of the formulae (I-1-g) to (I-2-g) shown above in which A, B, L, Q1, Q2, Q3, Q4, Q4, m, R6, R7 and Het are as defined above are obtained when compounds of the formulae (I-1-a) to (I-2-a) shown above in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined above are in each case (α) reacted with isocyanates or isothiocyanates of the formula (XII) R6—N═C═L (XII) in which R6 and L are as defined above, if appropriate in the presence of a diluent and if appropriate in the presence of a catalyst, or (β) reacted with carbamoyl chlorides or thiocarbamoyl chlorides of the formula (XII) in which L, R6 and R7 are as defined above, if appropriate in the presence of a diluent and if appropriate in the presence of an acid binder.","Furthermore, it has been found that the novel compounds of the formula (I) have, depending on the substituents, very good activity both as pesticides, preferably as insecticides and acaricides, and as herbicides and fungicides.","The formula (I) provides a general definition of the compounds according to the invention.","Preferred substituents or ranges of the radicals given in the formulae mentioned above and below are illustrated below: Het preferably represents m preferably represents the number 0 or 1, X preferably represents hydrogen, halogen, C1-C6-alkyl, C1-C6-alkoxy, C3-C6-alkenyloxy, nitro or cyano, Y preferably represents halogen, C1-C6-alkyl, C1-C6-halogenoalkyl, C1-C6-alkoxy, C1-C6-halogenoalkoxy or represents the groups V1 preferably represents hydrogen, halogen, C1-C12-alkyl, C1-C6-alkoxy, C1-C6-alkylthio, C1-C6-alkylsulphinyl, C1-C6-alkylsulphonyl, C1-C4-halogenoalkyl, C1-C4-halogenoalkoxy, nitro, cyano or represents phenyl, phenoxy, phenoxy-C1-C4-alkyl, phenyl-C1-C4-alkoxy, phenylthio-C1-C4-alkyl or phenyl-C1-C4-alkylthio, each of which is optionally mono- or polysubstituted by halogen, C1-C6-alkyl, C1-C6-alkoxy, C1-C4-halogenoalkyl, C1-C4-halogenoalkoxy, nitro or cyano, V2 and V3 independently of one another preferably represent hydrogen, halogen, C1-C6-alkyl, C1-C6-alkoxy, C1-C4-halogenoalkyl or C1-C4-halogenoalkoxy, V1 and V2 jointly together with the carbon atoms to which they are attached preferably represent an optionally C1-C4-alkyl- or halogen-substituted 5- or 6-membered cycle in which optionally one or two carbon atoms may be replaced by oxygen, sulphur or nitrogen, A preferably represents hydrogen or in each case optionally halogen-substituted C1-C12-alkyl, C3-C8-alkenyl, C1-C6-alkoxy-C1-C4-alkyl, optionally halogen-, C1-C4-alkyl- or C1-C4-alkoxy-substituted C3-C8-cycloalkyl or C3-C6-cycloalkyl-C1-C4-alkyl, in which optionally one or two not directly adjacent ring members are replaced by oxygen and/or sulphur, or represents in each case optionally halogen-, C1-C6-alkyl-, C1-C6-halogenoalkyl-, C1-C6-alkoxy-, C1-C6-halogenoalkoxy-, cyano- or nitro-substituted phenyl, benzyl, hetaryl having 5 or 6 ring atoms (for example furanyl, pyridyl, imidazolyl, triazolyl, pyrazolyl, pyrimidyl, thiazolyl or thienyl) or hetaryl-C1-C4-alkyl having 5 or 6 ring atoms (for example pyridyl-, pyrimidyl- or thiazolyl-C1-C4-alkyl), B preferably represents hydrogen or C1-C6-alkyl, A, B and the carbon atom to which they are attached preferably represent saturated C3-C11-cycloalkyl or unsaturated C5-C10-cycloalkyl in which optionally one ring member is replaced by oxygen or sulphur and which are optionally mono- or disubstituted by C1-C6-alkyl, C3-C8-cycloalkyl, C1-C6-halogenoalkyl, C1-C6-alkoxy, C1-C6-alkylthio, halogen or phenyl, A and Q1 together preferably represent C3-C6-alkanediyl which is optionally mono- or disubstituted by identical or different substituents from the group consisting of C1-C4-alkyl and C1-C4-alkoxy, Q1 preferably represents hydrogen, C1-C6-alkyl, C1-C6-alkoxy-C1-C2-alkyl, optionally fluorine-, chlorine-, C1-C4-alkyl-, C1-C2-halogenoalkyl- or C1-C4-alkoxy-substituted C3-C8-cycloalkyl in which optionally one methylene group is replaced by oxygen or sulphur or in each case optionally halogen-, C1-C4-alkyl-, C1-C4-alkoxy-, C1-C2-halogenoalkyl-, C1-C2-halogenoalkoxy-, cyano- or nitro-substituted phenyl, pyridyl, thienyl, thiazolyl, phenyl-C1-C4-alkyl, pyridyl -C11-C2-alkyl or thiazolyl-C1-C2-alkyl, Q2, Q3, Q4 independently of one another preferably represent hydrogen or C1-C4-alkyl, Q1 and Q2 together with the carbon atom to which they are attached preferably represent optionally C1-C6-alkyl-, C1-C6-alkoxy- or C1-C2-halogenoalkyl-substituted C3-C7-cycloalkyl in which optionally one ring member is replaced by oxygen or sulphur, G preferably represents hydrogen (a) or represents one of the groups in particular (a), (b), (c) or (g), in which E represents a metal ion or an ammonium ion, L represents oxygen or sulphur and M represents oxygen or sulphur, R1 preferably represents in each case optionally halogen-substituted C1-C20-alkyl, C2-C20-alkenyl, C1-C8-alkoxy-C1-C8-alkyl, C1-C8-alkylthio-C1-C8-alkyl, poly-C1-C8-alkoxy-C1-C8-alkyl or optionally halogen-, C1-C6-alkyl- or C1-C6-alkoxy-substituted C3-C8-cycloalkyl, in which optionally one or more (preferably one or two) not directly adjacent ring members are replaced by oxygen and/or sulphur, represents optionally halogen-, cyano-, nitro-, C1-C6-alkyl, C1-C6-alkoxy-, C1-C6-halogenoalkyl-, C1-C6-halogenoalkoxy-, C1-C6-alkylthio or C1-C6-alkylsulphonyl-substituted phenyl, represents optionally halogen-, nitro-, cyano-, C1-C6-alkyl-, C1-C6-alkoxy-, C1-C6-halogenoalkyl- or C1-C6-halogenoalkoxy-substituted phenyl-C1-C6-alkyl, represents optionally halogen-, C1-C6-alkyl- or trifluoromethyl-substituted 5- or 6-membered hetaryl (for example pyrazolyl, thiazolyl, pyridyl, pyrimidyl, furanyl or thienyl), represents optionally halogen- or C1-C6-alkyl-substituted phenoxy-C1-C6-alkyl or represents optionally halogen-, amino- or C1-C6-alkyl-substituted 5- or 6-membered hetaryloxy-C1-C6-alkyl (for example pyridyloxy-C1-C6-alkyl, pyrimidyloxy-C1-C6-alkyl or thiazolyloxy- C1-C6-alkyl), R2 preferably represents in each case optionally halogen-substituted C1-C20-alkyl, C2-C20-alkenyl, C1-C8-alkoxy-C2-C8-alkyl, poly-C1-C8-alkoxy-C2-C8-alkyl, represents optionally halogen-, C1-C6-alkyl- or C1-C6-alkoxy-substituted C3-C8-cycloalkyl or represents in each case optionally halogen-, cyano-, nitro-, C1-C6-alkyl-, C1-C6-alkoxy-, C1-C6-halogenoalkyl- or C1-C6-halogenoalkoxy-substituted phenyl or benzyl, R3 preferably represents optionally halogeno-substituted C1-C8-alkyl or represents in each case optionally halogen-, C1-C6-alkyl-, C1-C6-alkoxy-, C1-C4-halogenoalkyl-, C1-C4-halogenoalkoxy-, cyano- or nitro-substituted phenyl or benzyl, R4 and R5 independently of one another represent in each case optionally halogen-substituted C1-C8-alkyl, C1-C8-alkoxy, C1-C8-alkylamino, di-(C1-C8-alkyl)amino, C1-C8-alkylthio, C2-C8-alkenylthio, C3-C7-cycloalkylthio or represent in each case optionally halogen-, nitro-, cyano-, C1-C4-alkoxy-, C1-C4-halogenoalkoxy-, C1-C4-alkylthio-, C1-C4-halogenoalkylthio-, C1-C4-alkyl- or C1-C4-halogenoalkyl-substituted phenyl, benzyl, phenoxy or phenylthio, R6 and R7 independently of one another preferably represent hydrogen, represent in each case optionally halogen-substituted C1-C8-alkyl, C3-C8-cycloalkyl, C1-C8-alkoxy, C3-C8-alkenyl, C1-C8-alkoxy-C1-C8-alkyl, represent optionally halogen-, C1-C8-halogenoalkyl-, C1-C8-alkyl- or C1-C8-alkoxy-substituted phenyl, optionally halogen-, C1-C8-alkyl-, C1-C8-halogenoalkyl- or C1-C8-alkoxy-substituted benzyl or together with the nitrogen to which they are attached represent an optionally C1-C4-alkyl-substituted C3-C6-alkylene radical in which optionally one carbon atom is replaced by oxygen or sulphur.","In the radical definitions mentioned as being preferred, halogen represents fluorine, chlorine, bromine and iodine, in particular fluorine and chlorine, also as substituent, such as, for example, in halogenoalkyl.","Het particularly preferably represents m particularly preferably represents the number 0 or 1, X particularly preferably represents hydrogen, chlorine, bromine or C1-C4-alkyl, Y particularly preferably represents chlorine, bromine, C1-C4-alkyl, C1-C4-halogenoalkyl, C1-C4-alkoxy, C1-C4-halogenoalkoxy or the group V1 particularly preferably-represents hydrogen, fluorine, chlorine, C1-C6-alkyl, C1-C4-alkoxy, C1-C2-halogenoalkyl, C1-C2-halogenoalkoxy, nitro, cyano or phenoxy which is optionally mono- or disubstituted by fluorine, chlorine, bromine, C1-C4-alkyl, C1-C4-alkoxy, C1-C2-halogenoalkyl, C1-C2-halogenoalkoxy, nitro or cyano, V2 particularly preferably represents hydrogen, fluorine, chlorine, bromine, C1-C4-alkyl, C1-C4-alkoxy, C1-C2-halogenoalkyl or C1-C2-halogenoalkoxy, V1 and V2 jointly, together with the carbon atoms to which they are attached, particularly preferably represent an optionally fluorine- or methyl-substituted 5- or 6-membered cycle in which optionally one or two carbon atoms may be replaced by oxygen, A particularly preferably represents hydrogen, represents C1-C8-alkyl or C1-C4-alkoxy-C1-C2-alkyl, each of which is optionally mono- to trisubstituted fluorine, represents C5-C6-cycloalkyl or C3-C6-cycloalkyl-C1-C2-alkyl in which optionally one ring member is replaced by oxygen or sulphur, each of which radicals is optionally mono- or disubstituted by fluorine, chlorine, methyl, ethyl or methoxy, or represent phenyl or benzyl, each of which is optionally mono- or disubstituted by fluorine, chlorine, bromine, C1-C4-alkyl, C1-C2-halogenoalkyl, C1-C4-alkoxy or C1-C2-halogenoalkoxy, B particularly preferably represents hydrogen or C1-C4-alkyl, A, B and the carbon atom to which they are attached particularly preferably represent saturated C5-C6-cycloalkyl in which optionally one ring member is replaced by oxygen and which is optionally monosubstituted by C1-C4-alkyl, trifluoromethyl or C1-C4-alkoxy, with the proviso that in this case Q1 represents only hydrogen or C1-C4-alkyl, A and Q1 together particularly preferably represent C3-C4-alkanediyl which is optionally mono- or disubstituted by methyl, ethyl, methoxy or ethoxy, Q1 particularly preferably represents hydrogen, C1-C4-alkyl, C1-C4-alkoxy-C1-C2-alkyl, or optionally methyl- or methoxy-substituted C3-C6-cycloalkyl, in which optionally one methylene group is replaced by oxygen, or phenyl or benzyl, each of which is optionally mono- or disubstituted by fluorine, chlorine, bromine, C1-C4-alkyl, C1-C4-alkoxy, trifluoromethyl or trifluoromethoxy, Q2, Q3, Q4 independently of one another particularly preferably represent hydrogen, methyl or ethyl, Q1 and Q2 together with the carbon to which they are attached particularly preferably represent optionally C1-C4-alkyl- or C1-C4-alkoxy-substituted saturated C5-C6-cycloalkyl in which optionally one ring member is replaced by oxygen, with the proviso that in this case A only represents hydrogen or C1-C4-alkyl, G particularly preferably represents hydrogen (a) or represents one of the groups (g), in particular (a), (b) or (c) in which E represents a metal ion or an ammonium ion, L represents oxygen or sulphur and M represents oxygen or sulphur, R1 particularly preferably represents C1-C16-alkyl, C2-C16-alkenyl, C1-C4-alkoxy-C1-C2-alkyl, C1-C4-alkylthio-C1-C2-alkyl, each of which is optionally mono- to pentasubstituted by fluorine or chlorine, or C3-C6-cycloalkyl in which optionally one or two not directly adjacent ring members are replaced by oxygen and/or sulphur and which is optionally mono- or di substituted by fluorine, chlorine, C1-C4-alkyl or C1-C5-alkoxy, particularly preferably represents phenyl which is optionally mono- or disubstituted by fluorine, chlorine, bromine, cyano, nitro, C1-C4-alkyl, C1-C4-alkoxy, trifluoromethyl or trifluoromethoxy, particularly preferably represents pyridyl or thienyl, each of which is optionally monosubstituted by fluorine, chlorine, bromine, methyl, ethyl or trifluoromethyl, R2 particularly preferably represents C1-C16-alkyl, C2-C16-alkenyl or C1-C4-alkoxy-C2-C4-alkyl, particularly preferably represents C3-C6-cycloalkyl which is optionally mono- or disubstituted by methyl, ethyl or methoxy, particularly preferably represents phenyl or benzyl, each of which is optionally mono- or disubstituted by fluorine, chlorine, bromine, cyano, nitro, C1-C4-alkyl, C1-C4-alkoxy, trifluoromethyl or trifluoromethoxy, R3 particularly preferably represents C1-C4-alkyl which is optionally mono- to pentasubstituted by fluorine or represents phenyl which is optionally mono- or disubstituted by fluorine, chlorine, bromine, C1-C4-alkyl, C1-C4-alkoxy, trifluoromethyl, trifluoromethoxy, cyano or nitro, R4 particularly preferably represents C1-C6-alkyl, C1-C6-alkoxy, C1-C6-alkylamino, di-(C1-C6-alkyl)amino, C1-C6-alkylthio, or represents phenyl, benzyl, phenoxy or phenylthio, each of which is optionally mono- or disubstituted by fluorine, chlorine, bromine, nitro, cyano, C1-C3-alkoxy, trifluoromethoxy, C1-C3-alkyl or trifluoromethyl, R5 particularly preferably represents C1-C4-alkyl, C1-C4-alkoxy or C1-C4-alkylthio, R6 particularly preferably represents hydrogen, C1-C6-alkyl, C3-C6-cycloalkyl, C1-C6-alkoxy, C3-C6-alkenyl, C1-C6-alkoxy-C1-C6-alkyl, represents phenyl which is optionally mono- or disubstituted by fluorine, chlorine, bromine, trifluoromethyl, C1-C4-alkyl or C1-C4-alkoxy, represents benzyl which is optionally substituted by fluorine, chlorine, bromine, methyl, ethyl, trifluoromethyl or methoxy, R7 particularly preferably represents hydrogen, C1-C6-alkyl or C3-C6-alkenyl, R6 and R7 together, with nitrogen atom to which they are attached particularly preferably represent a C5-C6-alkylene radical in which optionally one methylene group is replaced by oxygen or sulphur and which is optionally mono- or disubstituted by methyl or ethyl.","In the radical definitions mentioned as being particularly preferred, halogen represents fluorine, chlorine and bromine, in particular fluorine and chlorine, especially fluorine, including as substituent, such as, for example, in halogenoalkyl.","Het very particularly preferably represents m very particularly preferably represents the number 0 or 1, X very particularly preferably represents hydrogen, chlorine, bromine, methyl, ethyl, n-propyl or isopropyl, Y very particularly preferably represents the group V1 very particularly preferably represents hydrogen, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, methoxy, ethoxy, n-propoxy, isopropoxy, trifluoromethyl, trifluoromethoxy or 4-chlorophenoxy, V2 very particularly preferably represents hydrogen, fluorine, chlorine, bromine, methyl, methoxy or trifluoromethyl, A very particularly preferably represents hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, methoxymethyl, ethoxymethyl, phenyl or cyclohexyl, B very particularly preferably represents hydrogen, methyl or ethyl, A, B and the carbon atom to which they are attached very particularly preferably represent saturated C5-C6-cycloalkyl in which optionally one ring member is replaced by oxygen and which is optionally monosubstituted by methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, trifluoromethyl, methoxy, ethoxy, n-propoxy, n-butoxy or isobutoxy, with the proviso that in this case Q1 only represents hydrogen, A and Q1 together very particularly preferably represent C3-C4-alkanediyl, Q1 very particularly preferably represents hydrogen, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, cyclopentyl, cyclohexyl or 4-chlorophenyl, Q2, Q3, Q4 independently of one another very particularly preferably represent hydrogen, methyl or ethyl, Q1 and Q2 together with the carbon atom to which they are attached very particularly preferably represent optionally methyl-, ethyl-, methoxy-, ethoxy-, n-propoxy- or n-butoxy-substituted saturated C5-C6-cycloalkyl in which optionally one ring member is replaced by oxygen, with the proviso that in this case A only represents hydrogen, G very particularly preferably represents hydrogen (a) or represents one of the groups (g), in particular (a), (b) or (c), in which E represents a metal ion or an ammonium ion, L represents oxygen or sulphur and M represents oxygen or sulphur, R1 very particularly preferably represents in each case optionally fluorine or chlorine-substituted C1-C8-alkyl, C2-C8-alkenyl, C1-C2-alkoxy-C1-C2-alkyl, C1-C2-alkylthio-C1-C2-alkyl or cyclopropyl or cyclohexyl, each of which is optionally monosubstituted by fluorine, chlorine, methyl, ethyl or methoxy, very particularly preferably represents phenyl which is optionally monosubstituted by fluorine, chlorine, bromine, cyano, nitro, methyl, ethyl, isopropyl, tert-butyl, methoxy, trifluoromethyl or trifluoromethoxy, very particularly preferably represents thienyl or pyridyl, each of which is optionally monosubstituted by fluorine, chlorine, bromine or methyl, R2 very particularly preferably represents C1-C8-alkyl, C2-C8-alkenyl or C1-C4-alkoxy-C2-alkyl, very particularly preferably represents C3-C6-cycloalkyl which is optionally monosubstituted by methyl, ethyl or methoxy, very particularly preferably represents phenyl or benzyl, each of which is optionally monosubstituted by fluorine, chlorine, cyano, nitro, methyl, ethyl, isopropyl, tert-butyl, methoxy, trifluoromethyl or trifluoromethoxy, R3 very particularly preferably represents methyl or ethyl, each of which is optionally trisubstituted by fluorine or phenyl which is optionally monosubstituted by fluorine, chlorine, bromine, methyl, tert-butyl, methoxy, trifluoromethyl, trifluoromethoxy, cyano or nitro, R4 very particularly preferably represents C1-C4-alkyl, C1-C4-alkoxy, C1-C4-alkylamino, di-(C1-C4-alkyl)amino, C1-C4-alkylthio or represents phenyl, phenoxy or phenylthio, each of which is optionally monosubstituted by fluorine, chlorine, bromine, nitro, cyano, C1-C2-alkoxy, trifluoromethoxy or C1-C3-alkyl, R5 very particularly preferably represents methyl, ethyl, methoxy, ethoxy, methylthio or ethylthio, R6 very particularly preferably represents hydrogen, represents C1-C4-alkyl, C3-C6-cycloalkyl, C1-C4-alkoxy, C3-C4-alkenyl or C1-C4-alkoxy-C1-C4-alkyl, R7 very particularly preferably represents hydrogen, C1-C4-alkyl or C3-C4-alkenyl, R6 and R7 together with the nitrogen atom to which they are attached very particularly preferably represent a C5-alkylene radical in which optionally one methylene group is replaced by oxygen.","Het most preferably represents m most preferably represents the number 0 or 1, X most preferably represents hydrogen, methyl or ethyl, Y most preferably represents the group V1 most preferably represents hydrogen, chlorine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, trifluoromethyl or represents 4-chloro-phenoxy, A most preferably represents hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, phenyl or cyclohexyl, B most preferably represents hydrogen, methyl or ethyl, A, B and the carbon atom to which they are attached most preferably represent saturated C5-C6-cycloalkyl in which optionally one ring member is replaced by oxygen and which is optionally monosubstituted by methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, methoxy, ethoxy, n-propoxy, n-butoxy or isobutoxy, with the proviso that in this case Q1 only represents hydrogen, A and Q1 together most preferably represent C3-C4-alkanediyl, Q1 most preferably represents hydrogen, methyl, ethyl, n-propyl, isopropyl or 4-chlorophenyl, Q2 most preferably represents hydrogen, methyl or ethyl, Q3 most preferably represents hydrogen, Q4 most preferably represents hydrogen, Q1 and Q2 together with the carbon atom to which they are attached most preferably represent saturated C5-C6-cycloalkyl, with the proviso that in this case A only represents hydrogen, G most preferably represents hydrogen (a) or represents one of the groups in which L represents oxygen and M represents oxygen or sulphur, R1 most preferably represents C1-C8-alkyl, C1-C2-alkoxy-C1-C2-alkyl or cyclopropyl which is optionally monosubstituted by chlorine, represents phenyl which is optionally monosubstituted by chlorine, represents pyridyl which is optionally monosubstituted by chlorine, R2 most preferably represents C1-C8-alkyl, represents phenyl or benzyl, R3 most preferably represents methyl or ethyl, R6 and R7 together with the nitrogen atom to which they are attached most preferably represent a C5-alkylene radical in which optionally one methylene group is replaced by oxygen.","The general or preferred radical definitions or illustrations given above can be combined with one another as desired, i.e.","including combinations between the respective ranges and preferred ranges.","They apply to the end products and, correspondingly, to precursors and intermediates.","Preference according to the invention is given to the compounds of the formula (I) which contain a combination of the meanings given above as being preferred (preferable).","Particular preference according to the invention is given to the compounds of the formula (I) which contain a combination of the meanings given above as being particularly preferred.","Very particular preference according to the invention is given to the compounds of the formula (I) which contain a combination of the meanings given above as being very particularly preferred.","Most preference according to the invention is given to the compounds of the formula (I) which contain a combination of the meanings given above as being most preferred.","Particular preference is given to compounds of the formula (I) in which G represents hydrogen.","Saturated or unsaturated hydrocarbon radicals, such as alkyl or alkenyl, can in each case be straight-chain or branched as far as this is possible, including in combination with heteroatoms, such as, for example, in alkoxy.","Unless indicated otherwise, optionally substituted radicals can be mono- or poly-substituted, where in the case of polysubstitution the substituents can be identical or different.","Using, for example, according to process (A) 1-[4-(5-methyl)-2-(4-chloro-phenyl)-thiazolyl]-3-(1-ethoxycarbonyl-cyclohexyl)-propanone as starting material, the course of the process according to the invention can be represented by the reaction scheme below: Using, for example, according to process (B) 1-[4-(5-methyl)-2-(4-chloro-phenyl)-thiazolyl]-4-(l-ethoxycarbonyl-cyclohexyl)-butan-2-one as starting material, the course of the process according to the invention can be represented by the following reaction scheme: Using, for example, according to process (Dα) 2-[4-(5-methyl-2-(3-chloro-phenyl)-thiazolyl]-5,5-dimethylcyclohexane-1,3-dione and pivaloyl chloride as starting materials, the course of the process according to the invention can be represented by the following reaction scheme: Using, for example, according to process (Dβ) 2-[4-(5-ethyl-2-(4-methoxy-phenyl))-thiazolyl]-5,5-dimethyl-cyclohexane-1,3-dione and acetic anhydride as starting materials, the course of the process according to the invention can be represented by the reaction scheme below: Using, for example, according to process (E) 2-[4-(5-methyl-2-phenyl)-thiazolyl]-5,5-dimethyl-cyclohexane-1,3-dione and ethoxyethyl chloroformate as starting materials, the course of the process according to the invention can be represented by the reaction scheme below: Using, for example, according to process (F), 2-[4-(5-methyl-2-(4-fluorophenyl))-thiazolyl]-4,4-dimethyl-cyclopentane-1,3-dione and methyl chloromonothioformate as starting materials, the course of the reaction can be represented as shown below: Using, for example, according to process (G) 2-[4-(5-methyl-3-(4-methyl-phenyl)-thiazolyl]-5,5-dimethyl-cyclohexane-1,3-dione and methanesulphonyl chloride as starting materials, the course of the reaction can be represented by the reaction scheme below: Using, for example, according to process (H) 2-[4-(5-methyl-2-phenyl)-thiazolyl]-4,4-dimethyl-cyclopentane-1,3-dione and 2,2,2-trifluoroethyl methanethio-phosphonyl chloride as starting materials, the course of the reaction can be represented by the reaction scheme below: Using, for example, according to process (I) 2-[4-(5-methyl-2-(4-trifluornethyl-phenyl))-thiazolyl]-5,5-dimethyl-cyclohexane-1,3-dione and NaOH as components, the course of the process according to the invention can be represented by the reaction scheme below: Using, for example, according to process (Jα) 2-[4-(5-methyl-2-(3-trifluoromethyl-phenyl))-thiazolyl]-4,4-dimethyl-cyclohexane-1,3-dione and ethyl isocyanate as starting materials, the course of the reaction can be represented by the reaction scheme below: Using, for example, according to process (Jβ) 2-[4-(5-methyl-2-phenyl)-thiazolyl]-5,5-dimethyl-cyclohexyl-1,3-dione and dimethylcarbamoyl chloride as starting materials, the course of the reaction can be represented by the scheme below: The ketocarboxylic esters of the formula (II) in which A, B, Het, Q1, Q2 and R8 are as defined above, which are required as starting materials in the above process (A), are novel.","They can be prepared by methods known in principle.","The 5-hetaryl-4-ketocarboxylic esters of the formula (II) are obtained, for example, when 5-hetaryl-4-ketocarboxylic acids of the formula (XIV) in which A, B, Het, Q1 and Q2 are as defined above, are esterified (cf., for example, Organikum, 15th edition, Berlin, 1977,page 499) or alkylated (see Preparation Example).","The 5-hetaryl4-ketocarboxylic acids of the formula (XIV) in which A, B, Het, Q1 and Q2 are as defined above, are novel; they can however be prepared by methods known in principle (see Preparation Example).","The 5-hetaryl4-ketocarboxylic acids of the formula (XIV) are obtained, for example, when 2-hetaryl-3-oxo-adipic esters of the formula (XV) in which A, B, Het, Q1 and Q2 are as defined above and R8 and R8′ represent alkyl (in particular C1-C8-alkyl) are decarboxylated, if appropriate in the presence of a diluent and if appropriate in the presence of a base or an acid (cf., for example Organikum, 15th edition, Berlin, 1977,pages 519-521).","The compounds of the formula (XV) in which A, B, Het, Q1, Q2, R8 and R8′ are as defined above, or, if the compound of the formula (XVII) was used as starting material for the preparation, in which case R8 represents hydrogen, are novel.","The compounds of the formula (XV) are obtained, for example, when dicarboxylic monoester chlorides of the formula (XVI), in which A, B, Q1, Q2 and R8 are as defined above and Hal represents chlorine or bromine, or carboxylic anhydrides of the formula (XVII) in which A, B, Q1 and Q2 are as defined above, are acylated with a hetarylacetic ester of the formula (XVIII) in which Het and R8′ are as defined above, in the presence of a diluent and in the presence of a base (cf., for example, M. S. Chambers, E. J. Thomas, D. J. Williams, J. Chem.","Soc.","Chem.","Commun., (1987), 1228,cf.","also the Preparation Examples).","Some of the compounds of the formulae (XVI) and (XVII) are known compounds of organic chemistry, and/or they can be prepared in a simple manner by methods known in principle.","The ketocarboxylic esters of the formula (III) in which A, B, Het, Ql, Q2, Q3, Q4 and R8 are as defined above, which are required as starting materials for the above process (B), are novel.","They can be prepared by methods known in principle.","The 6-hetaryl-5-ketocarboxylic esters of the formula (III) are obtained, for example, when 6-hetaryl-5-ketocarboxylic acids of the formula (XIX) in which A, B, Het, Q1, Q2, Q3 and Q4 are as defined above, are esterified (cf., for example, Organikum, 15th edition, Berlin, 1977,page 499) or alkylated (see Preparation Example).","The 6-aryl-5-ketocarboxylic acids of the formula (XIX) in which A, B, Het, Q1, Q2, Q3 and Q4 are as defined above are novel.","They can be prepared by methods known in principle, for example by hydrolysing and decarboxylating substituted 2-hetaryl-3-oxo-heptanedioic esters of the formula (XX) in which A, B, Het, Q1, Q2, Q3 and Q4 are as defined above and R8 and R8′ represent alkyl (preferably C1-C6-alkyl), if appropriate in the presence of a diluent and if appropriate in the presence of a base or an acid (cf., for example, Organikum, 15th edition, Berlin, 1977,pages 519-521).","The compounds of the formula (XX) in which A, B, Het, Q1, Q2, Q3, Q4, R8 and R8′ are as defined above, or, if the compound of the formula (XXII) was used as starting material for the preparation, in which case R8 represents hydrogen, are novel.","The compounds of the formula (XX) are obtained, for example, when dicarboxylic esters of the formula (XXI), or dicarboxylic anhydrides of the formula (XXII) in which A, B, Q1, Q2, Q3, Q4 and R8 are as defined above, are condensed with a substituted hetarly acetic ester of the formula (XVII) in which Het and R8′ are as defined above, in the presence of a diluent and in the presence of a base.","Some of the compounds of the formulae (XXI) and (XXII) are commercially available, some are known, and/or they can be prepared by known processes.","Some of the compounds of the formula (XVIII) are novel; however, they can be prepared by processes known in principle: 1.C.","S. Rooney et al.","J. Med.","Chem.","1983, 26, 700-714.2.M.","S. Malamas et al.","J. Med.","Chem.","1996, 39, 237-245.3.J.","L. Collins et al.","J. Med.","Chem.","1998, 41, 5037-5054.4.EP-A-0 177 353.5.EP-A-0 368 592.6.NL-A-6614130.The acyl halides of the formula (VI), carboxylic anhydrides of the formula (V), chloroformic esters or chloroformic thioesters of the formula (VI), chloromonothio-formic esters or chlorodithioformic esters of the formula (V), sulphonyl chlorides of the formula (VII), phosphorus compounds of the formula (IX) and metal hydroxides, metal alkoxides or amines of the formulae (X) and (XI) and isocyanates of the formula (XII) and carbamoyl chlorides or thiocarbamoyl chlorides of the formula (XI) furthermore required as starting materials for carrying out the processes (C), (D), (E), (F), (G), (H), and (I) according to the invention are generally known compounds of organic or inorganic chemistry.","Some of the compounds of the formulae (XVI), (XVII) and (XXI) are cornmercially available, some are known, and/or they can be prepared by methods known in principle.","The process (A) is characterized in that compounds of the formula (II) in which A, B, Q1, Q2, Het and R8 are as defined above are subjected to an intramolecular condensation in the presence of a base.","Suitable for use as diluents in the process (A) according to the invention are all inert organic solvents.","Preference is given to using hydrocarbons, such as toluene and xylene, furthermore ethers, such as dibutyl ether, tetrahydrofuran, dioxane, glycol dimethyl ether and diglycol dimethyl ether, moreover polar solvents, such as dimethyl sulphoxide, sulpholane, dimethylformamide, dimethylacetamide and N-methyl-pyrrolidone, and also alcohols, such as methanol, ethanol, propanol, iso-propanol, butanol, iso-butanol and tert-butanol.","Suitable bases (deprotonating agents) for carrying out the process (A) according to the invention are all customary proton acceptors.","Preference is given to using alkali metal and alkaline earth metal oxides, hydroxides and carbonates, such as sodium hydroxide, potassium hydroxide, magnesium oxide, calcium oxide, sodium carbonate, potassium carbonate and calcium carbonate, which can also be used in the presence of phase-transfer catalysts, such as, for example, triethylbenzylammonium chloride, tetrabutylamrnmonium bromide, Adogen 464 (=methyltrialkyl(C8-C10)ammonium chloride) or TDA 1 (=tris-(methoxyethoxyethyl)-amine).","It is furthermore possible to use alkali metals such as sodium or potassium.","Furthermore, it is possible to use alkali metal and alkaline earth metal amides and hydrides, such as sodium amide, sodium hydride and calcium hydride, and, moreover, also alkali metal alkoxides, such as sodium methoxide, sodium ethoxide and potassium tert-butoxide.","When carrying out the process (A) according to the invention, the reaction temperatures can be varied within a relatively wide range.","In general, the process is carried out at temperatures between −80° C. and 180° C., preferably between −50° C. and 120° C. The process (A) according to the invention is generally carried out under atmospheric pressure.","When carrying out the process (A) according to the invention, the reaction components of the formula (II) and the deprotonating bases are generally employed in approximately double-equimolar amounts.","However, it is also possible to use a relatively large excess (up to 3 mol) of one or the other component.","The process (B) is characterized in that compounds of the formula (III) in which A, B, Q1, Q2, Q3, Q4, Het and R8 are as defined above are subjected to an intramolecular condensation in the presence of a diluent and in the presence of a base.","Suitable for use as diluents in the process (B) according to the invention are all inert organic solvents.","Preference is given to using hydrocarbons, such as toluene and xylene, furthermore ethers, such as dibutyl ether, tetrahydrofuran, dioxane, glycol dimethyl ether and diglycol dimethyl ether, moreover polar solvents, such as dimethyl sulphoxide, sulpholane, dimethylformamide and N-methyl-pyrrolidone.","It is furthermore possible to use alcohols, such as methanol, ethanol, propanol, iso-propanol, butanol, iso-butanol and tert-butanol.","Suitable bases (deprotonating agents) for carrying out the process (B) according to the invention are all customary proton acceptors.","Preference is given to using alkali metal and alkaline earth metal oxides, hydroxides and carbonates, such as sodium hydroxide, potassium hydroxide, magnesium oxide, calcium oxide, sodium carbonate, potassium carbonate and calcium carbonate, which can also be used in the presence of phase-transfer catalysts, such as, for example, triethylbenzylammonium chloride, tetrabutylammonium bromide, Adogen 464 (=methyltrialkyl(C8-C10)ammonium chloride) or TDA 1 (=tris-(methoxyethoxyethyl)-amine).","It is furthermore possible to use alkali metals such as sodium or potassium.","Furthermore, it is possible to use alkali metal and alkaline earth metal amides and hydrides, such as sodium amide, sodium hydride and calcium hydride, and, moreover, also alkali metal alkoxides, such as sodium methoxide, sodium ethoxide and potassium tert-butoxide.","When carrying out the process (B) according to the invention, the reaction temperatures can be varied within a relatively wide range.","In general, the process is carried out at temperatures between −80° C. and 180° C., preferably between −50° C. and 120° C. The process (B) according to the invention is generally carried out under atmospheric pressure.","When carrying out the process (B) according to the invention, the reaction components of the formula (III) and the deprotonating bases are generally employed in approximately equimolar amounts.","However, it is also possible to use a relatively large excess (up to 3 mol) of one or the other component.","The process (C-α) is characterized in that compounds of the formulae (I-1-a) to (I-2-a) are in each case reacted with carbonyl halides of the formula (IV), if appropriate in the presence of a diluent and if appropriate in the presence of an acid binder.","Suitable diluents for the process (C-60 ) according to the invention are all solvents which are inert to the acyl halides.","Preference is given to using hydrocarbons, such as benzine, benzene, toluene, xylene and tetraline, furthermore halogenated hydrocarbons, such as methylene chloride, chloroform, carbon tetrachloride, chlorobenzene and o-dichlorobenzene, moreover ketones, such as acetone and methyl isopropyl ketone, furthermore ethers, such as diethyl ether, tetrahydrofuran and dioxane, additionally carboxylic esters, such as ethyl acetate, nitriles, such as acetonitrile, and also strongly polar solvents, such as dimethylformamide, dimethylacetamide, dimethyl sulphoxide and sulpholane.","The hydrolytic stability of the acyl halide permitting, the reaction can also be carried out in the presence of water.","Suitable acid binders for the reaction according to the process (C-α) according to the invention are all customary acid acceptors.","Preference is given to using tertiary amines, such as triethylamine, pyridine, diazabicyclooctane (DABCO), diazabi-cycloundecene (DBU), diazabicyclononene (DBN), Hünig base or N,N-dimethylaniline, furthermore alkaline earth metal oxides, such as magnesium oxide and calcium oxide, moreover alkali metal and alkaline earth metal carbonates, such as sodium carbonate, potassium carbonate and calcium carbonate, and also alkali metal hydroxides, such as sodium hydroxide and potassium hydroxide.","The reaction temperatures in the process (C-α) according to the invention can be varied within a relatively wide range.","In general, the process is carried out at temperatures between −20° C. and +150° C., preferably between 0° C. and 100° C. When carrying out the process (C-α) according to the invention, the starting materials of the formulae (I-1-a) to (I-2-a) and the carbonyl halide of the formula (IV) are generally each employed in approximately equivalent amounts.","However, it is also possible to use a relatively large excess (up to 5 mol) of the carbonyl halide.","Work-up is carried out by customary methods.","The process (C-β) is characterized in that compounds of the formulae (I1-a) to (I2-a) are reacted with carboxylic anhydrides of the formula (V), if appropriate in the presence of a diluent and if appropriate in the presence of an acid binder.","Suitable diluents for use in the process (C-β) according to the invention are preferably those solvents which are also preferred when acyl halides are used.","Furthermore, it is also possible for a carboxylic anhydride employed in excess to act simultaneously as diluent.","Suitable acid binders, which are, if appropriate, added in the process (C-β), are preferably those acid binders which are also preferred when acyl halides are used.","The reaction temperatures in the process (C-β) according to the invention can be varied within a relatively wide range.","In general, the process is carried out at temperatures between −20° C. and +150° C., preferably between 0° C. and 100° C. When carrying out the process (C-β) according to the invention, the starting materials of the formulae (I-1-a) to (I2-a) and the carboxylic anhydride of the formula (V) are generally each employed in approximately equivalent amounts.","However, it is also possible to use a relatively large excess (up to 5 mol) of the carboxylic anhydride.","Work-up is carried out by customary methods.","In general, diluent and excess carboxylic anhydride and the carboxylic acid that is formed are removed by distillation or by washing with an organic solvent or with water.","The process (D) is characterized in that compounds of the formulae (I1-a) to (I2-a) are in each case reacted with chlorofornic esters or chloroformic thiolesters of the formula (VI), if appropriate in the presence of a diluent and if appropriate in the presence of an acid binder.","Suitable acid binders for the reaction according to process (D) according to the invention are all customary acid acceptors.","Preference is given to using tertiary amines, such as triethylamine, pyridine, DABCO, DBU, DBA, Hünig base and N,N-dimethyl-aniline, furthermore alkaline earth metal oxides, such as magnesium oxide and calcium oxide, moreover alkali metal and alkaline earth metal carbonates, such as sodium carbonate, potassium carbonate and calcium carbonate, and also alkali metal hydroxides, such as sodium hydroxide and potassium hydroxide.","Suitable diluents for the process (D) according to the invention are all solvents which are inert to the chloroformic esters or chloroformic thiolesters used.","Preference is given to using hydrocarbons, such as benzine, benzene, toluene, xylene and tetraline, furthermore halogenated hydrocarbons, such as methylene chloride, chloroform, carbon tetrachloride, chlorobenzene and o-dichlorobenzene, moreover ketones, such as acetone and methyl isopropyl ketone, furthermore ethers, such as diethyl ether, tetrahydrofuran and dioxane, additionally carboxylic esters, such as ethyl acetate, nitriles, such as acetonitrile, and also strongly polar solvents, such as dimethylformamide, dimethylacetamide, dimethyl sulphoxide and sulpholane.","When carrying out the process (D) according to the invention, the reaction temperatures can be varied within a relatively wide range.","If the process is carried out in the presence of a diluent and an acid binder, the reaction temperatures are generally between −20° C. and +100° C., preferably between 0° C. and 50° C. The process (D) according to the invention is generally carried out under atmospheric pressure.","When carrying out the process (D) according to the invention, the starting materials of the formulae (I-1-a) to (I2-a) and the appropriate chloroformic ester or chloroformic thiolester of the formula (VI) are generally each employed in approximately equivalent amounts.","However, it is also possible to use a relatively large excess (up to 2 mol) of one or the other component.","Work-up is carried out by customary methods.","In general, precipitated salts are removed and the reaction mixture that remains is concentrated by stripping off the diluent.","Process (E) according to the invention is characterized in that compounds of the formulae (I1-a) to (I-2-a) are in each case reacted with compounds of the formula (VII), in the presence of a diluent and, if appropriate, in the presence of an acid binder.","In the preparation process (E), about 1 mol of chloromonothioformic ester or chlorodithioformic ester of the formula (VII) is employed per mole of starting material of the formulae (I-1-a) to (I2-a), at from 0 to 120° C., preferably at from 20 to 60° C. Suitable diluents, which are added, if appropriate, are all inert polar organic solvents, such as nitrites, esters, ethers, amides, sulphones, sulphoxides, and also halogenoalkanes.","Preference is given to using acetonitrile, ethyl acetate, dimethyl sulphoxide, tetra-hydrofuran, dimethylformamide or methylene chloride.","If, in a preferred embodiment, the enolate salt of the compounds (I1-a) to (I-2-a) is prepared by adding strong deprotonating agents, such as, for example, sodium hydride or potassium tert-butoxide, the further addition of acid binders can be dispensed with.","If acid binders are used, these are customary inorganic or organic bases, for example sodium hydroxide, sodium carbonate, potassium carbonate, pyridine, triethylamine.","The reaction can be carried out at atmospheric pressure or under elevated pressure and is preferably carried out at atmospheric pressure.","Work-up is carried out by customary methods.","The process (F) according to the invention is characterized in that compounds of the formulae (I1-a) to (I2-a) are in each case reacted with sulphonyl chlorides of the formula (VII), if appropriate in the presence of a diluent and if appropriate in the presence of an acid binder.","In the preparation process (F), about 1 mol of sulphonyl chloride of the formula.","(VII) is employed per mole of starting material of the formulae (I-1-a) to (I2-a), at from −20 to 150° C., preferably at from 20 to 70° C. Suitable diluents which are added, if appropriate, are all inert polar organic solvents, such as nitrites, esters, ethers, amides, sulphones, sulphoxides, or halogenated hydrocarbons, such as methylene chloride.","Preference is given to using acetonitrile, ethyl acetate, dimethyl sulphoxide, tetra-hydrofuran, dimethylformamide, methylene chloride.","If, in a preferred embodiment, the enolate salt of the compounds (I1-a) to (I-2-a) is prepared by adding strong deprotonating agents (such as, for example, sodium hydride or potassium tert-butoxide), the further addition of acid binders can be dispensed with.","If acid binders are used, these are customary inorganic or organic bases, for example sodium hydroxide, sodium carbonate, potassium carbonate, pyridine, triethylamine.","The reaction can be carried out at atmospheric pressure or under elevated pressure and is preferably carried out at atmospheric pressure.","Work-up is carried out by customary methods.","The process (G) according to the invention is characterized in that compounds of the formulae (I1-a) to (I2-a) are in each case reacted with phosphorus compounds of the formula (IX), if appropriate in the presence of a diluent and if appropriate in the presence of an acid binder.","In the preparation process (G), to obtain compounds of the formulae (1-1-e) to (1-2-e), 1 to 2,preferably 1 to 1.3,mol of the phosphorus compound of the formula (IX) are employed per mole of the compounds (I1-a) to (I-2-a), at temperatures between −40° C. and 150° C., preferably between −10 and 110° C. Suitable diluents which are added, if appropriate, are all inert polar organic solvents, such as ethers, amides, nitrites, alcohols, sulphides, sulphones, sulphoxides, etc.","Preference is given to using acetonitrile, dimethyl sulphoxide, tetrahydrofuran, dimethylformamide, methylene chloride.","Suitable acid binders which are added, if appropriate, are customary inorganic or organic bases, such as hydroxides, carbonates or amines.","Examples which may be mentioned are sodium hydroxide, sodium carbonate, potassium carbonate, pyridine, triethylamine.","The reaction can be carried out at atmospheric pressure or under elevated pressure and is preferably carried out at atmospheric pressure.","Work-up is carried out by customary methods of organic chemistry.","The end products obtained are preferably purified by crystallization, chromatographic purification or by “incipient distillation”, i.e.","removal of the volatile components under reduced pressure.","The process (H) is characterized in that compounds of the formulae (I-1-a) to (I2-a) are reacted with metal hydroxides or metal alkoxides of the formula (X) or amines of the formula (XI), if appropriate in the presence of a diluent.","Suitable for use as diluents in the process (H) according to the invention are, preferably, ethers, such as tetrahydrofuran, dioxane, diethyl ether, or else alcohols, such as methanol, ethanol, isopropanol, and also water.","The process (H) according to the invention is generally carried out under atmospheric pressure.","The reaction temperatures are generally between −20° C. and 100° C., preferably between 0° C. and 50° C. The process (I) according to the invention is characterized in that compounds of the formulae (I-1-a) to (I2-a) are in each case reacted with compounds of the formula (XII) (I-α), if appropriate in the presence of a diluent and if appropriate in the presence of a catalyst, or (I-β) with compounds of the formula (XII), if appropriate in the presence of a diluent and if appropriate in the presence of an acid binder.","In the preparation process (I-α), about 1 mol of isocyanate of the formula (XII) is employed per mole of starting material of the formulae (I1-a) to (I2-a), at from 0 to 100° C., preferably at from 20 to 50° C. Suitable diluents which are added, if appropriate, are all inert organic solvents, such as ethers, amides, nitriles, sulphones, sulphoxides.","If appropriate, catalysts may be added to accelerate the reaction.","Very advantageously, the catalysts which are employed are organotin compounds, such as, for example, dibutyltin dilaurate.","The reaction is preferably carried out at atmospheric pressure.","In preparation process (I-β), about 1 mol of carbamoyl chloride of the formula (XIII) is reacted per mole of starting material of the formulae (I-1-a) to (I-2-a) at from −20 to 150° C., preferably at from 0 to 70° C. Suitable diluents which are added, if appropriate, are all inert polar organic solvents, such as nitrites, esters, ethers, amides, sulphones, sulphoxides or halogenated hydrocarbons.","Preference is given to using acetonitrile, ethyl acetate, dimethyl sulphoxide, tetrahydrofuran, dimethylformamide or methylene chloride.","If, in a preferred embodiment, the enolate salt of the compounds (I-1-a) to (I2-a) is prepared by adding strong deprotonating agents (such as, for example, sodium hydride or potassium tert-butoxide), the addition of further acid binders can be dispensed with.","The active compounds are suitable for controlling animal pests, in particular insects, arachnids and nematodes, which are encountered in agriculture, in forestry, in the protection of stored products and of materials, and in the hygiene sector, and have good plant tolerance and favourable toxicity to warm-blooded animals.","They may be preferably employed as plant protection agents.","They are active against normally sensitive and resistant species and against all or some stages of development.","The abovementioned pests include: From the order of the Isopoda, for example, Oniscus asellus, Armadillidium vulgare and Porcellio scaber.","From the order of the Diplopoda, for example, Blaniulus guttulatus.","From the order of the Chilopoda, for example, Geophilus carpophagus and Scutigera spp.","From the order of the Symphyla, for example, Scutigerella immaculata.","From the order of the Thysanura, for example, Lepisma saccharina.","From the order of the Collembola, for example, Onychiurus armatus.","From the order of the Orthoptera, for example, Acheta domesticus, Gryllotalpa spp., Locusta migratoria migratorioides, Melanoplus spp.","and Schistocerca gregaria.","From the order of the Blattaria, for example, Blatta orientalis, Periplaneta americana, Leucophaea maderae, Blattella germanica.","From the order of the Dermaptera, for example, Forficula auricularia.","From the order of the Isoptera, for example, Reticulitermes spp.","From the order of the Phthiraptera, for example, Pediculus humanus corporis, Haematopinus spp., Linognathus spp., Trichodectes spp.","and Damalinia spp.","From the order of the Thysanoptera, for example, Hercinothrips femoralis, Thrips tabaci, Thrips palmi and Frankliniella accidentalis.","From the order of the Heteroptera, for example, Eurygaster spp., Dysdercus intermedius, Piesma quadrata, Cimex lectularius, Rhodnius prolixus and Triatoma spp.","From the order of the Homoptera, for example, Aleurodes brassicae, Bemisia tabaci, Trialeurodes vaporariorum, Aphis gossypii, Brevicoryne brassicae, Cryptomyzus ribis, Aphis fabae, Aphis pomi, Eriosoma lanigerum, Hyalopterus arundinis, Phylloxera vastatrix, Pemphigus spp., Macrosiphum avenae, Myzus spp., Phorodon humuli, Rhopalosiphum padi, Empoasca spp., Euscelis bilobatus, Nephotettix cincticeps, Lecanium corni, Saissetia oleae, Laodelphax striatellus, Nilaparvata lugens, Aonidiella aurantii, Aspidiotus hederae, Pseudococcus spp.","and Psylla spp.","From the order of the Lepidoptera, for example, Pectinophora gossypiella, Bupalus piniarius, Cheimatobia brumata, Lithocolletis blancardella, Hyponomeuta padella, Plutella xylostella, Malacosoma neustria, Euproctis chrysorrhoea, Lymantria spp., Bucculatrix thurberiella, Phyllocnistis citrella, Agrotis spp., Euxoa spp., Feltia spp., Earias insulana, Heliothis spp., Mamestra brassicae, Panolis flammea, Spodoptera spp., Trichoplusia ni, Carpocapsa pomonella, Pieris spp., Chilo spp., Pyrausta nubilalis, Ephestia kuehniella, Galleria mellonella, Tineola bisselliella, Tinea pellionella, Hofmannophila pseudospretella, Cacoecia podana, Capua reticulana, Choristoneura fumiferana, Clysia ambiguella, Homona magnanima, Tortrix viridana, Cnaphalocerus spp., Oulema oryzae.","From the order of the Coleoptera, for example, Anobium punctatum, Rhizopertha dominica, Bruchidius obtectus, Acanthoscelides obtectus, Hylotrupes bajulus, Agelastica alni, Leptinotarsa decemlineata, Phaedon cochleariae, Diabrotica spp., Psylliodes chrysocephala, Epilachna varivestis, Atomaria spp., Oryzaephilus surinamensis, Anthonomus spp., Sitophilus spp., Otiorrhynchus sulcatus, Cosmopolites sordidus, Ceuthorrhynchus assimilis, Hypera postica, Dennestes spp., Trogoderma spp., Anthrenus spp., Attagenus spp., Lyctus spp., Meligethes aeneus, Ptinus spp., Niptus hololeucus, Gibbium psylloides, Tribolium spp., Tenebrio molitor, Agriotes spp., Conoderus spp., Melolontha melolontha, Amphimallon solstitialis, Costelytra zealandica and Lissorhoptrus oryzophilus.","From the order of the Hymenoptera, for example, Diprion spp., Hoplocampa spp., Lasius spp., Monomorium pharaonis and Vespa spp.","From the order of the Diptera, for example, Aedes spp., Anopheles spp., Culex spp., Drosophila melanogaster, Musca spp., Fannia spp., Calliphora erythrocephala, Lucilia spp., Chrysomyia spp., Cuterebra spp., Gastrophilus spp., Hyppobosca spp., Stomoxys spp., Oestrus spp., Hypoderma spp., Tabanus spp., Tannia spp., Bibio hortulanus, Oscinella frit, Phorbia spp., Pegomyia hyoscyami, Ceratitis capitata, Dacus oleae, Tipula paludosa, Hylemyia spp.","and Liriomyza spp.","From the order of the Siphonaptera, for example, Xenopsylla cheopis and Ceratophyllus spp.","From the class of the Arachnida, for example, Scorpio maurus, Latrodectus mactans, Acarus siro, Argas spp., Omithodoros spp., Dermanyssus gallinae, Eriophyes ribis, Phyllocoptruta oleivora, Boophilus spp., Rhipicephalus spp., Amblyomma spp., Hyalomma spp., Ixodes spp., Psoroptes spp., Chorioptes spp., Sarcoptes spp., Tarsonemus spp., Bryobia praetiosa, Panonychus spp., Tetranychus spp., Hemitarsonemus spp., Brevipalpus spp.","The phytoparasitic nematodes include, for example, Pratylenchus spp., Radopholus similis, Ditylenchus dipsaci, Tylenchulus semipenetrans, Heterodera spp., Globodera spp., Meloidogyne spp., Aphelenchoides spp., Longidorus spp., Xiphinema spp., Trichodorus spp., Bursaphelenchus spp.","If appropriate, the compounds according to the invention can, at certain concentrations or application rates, also be used as herbicides or microbicides, for example as fungicides, antimycotics and bactericides.","If appropriate, they can also be employed as intermediates or precursors for the synthesis of other active compounds.","All plants and plant parts can be treated in accordance with the invention.","Plants are to be understood as meaning in the present context all plants and plant populations such as desired and undesired wild plants or crop plants (including naturally occurring crop plants).","Crop plants can be plants which can be obtained by conventional plant breeding and optimization methods or by biotechnological and recombinant methods or by combinations of these methods, including the transgenic plants and inclusive of the plant cultivars protectable or not protectable by plant breeders rights.","Plant parts are to be understood as meaning all parts and organs of plants above and below the ground, such as shoot, leaf, flower and root, examples which may be mentioned being leaves, needles, stalks, stems, flowers, fruit bodies, fruits, seeds, roots, tubers and rhizomes.","The plant parts also include harvested material, and vegetative and generative propagation material, for example cuttings, tubers, rhizomes, offsets and seeds.","Treatment according to the invention of the plants and plant parts with the active compounds is carried out directly or by allowing the compounds to act on the surroundings, environment or storage space by the customary treatment methods, for example by immersion, spraying, evaporation, fogging, scattering, painting on and, in the case of propagation material, in particular in the case of seeds, also by applying one or more coats.","The active compounds can be converted to the customary formulations, such as solutions, emulsions, wettable powders, suspensions, powders, dusts, pastes, soluble powders, granules, suspension-emulsion concentrates, natural and synthetic materials impregnated with active compound and microencapsulations in polymeric substances.","These formulations are produced in a known manner, for example by mixing the active compounds with extenders, that is, liquid solvents, and/or solid carriers, optionally with the use of surfactants, that is emulsifiers and/or dispersants, and/or foam-formers.","If the extender used is water, it is also possible to employ for example organic solvents as auxiliary solvents.","Essentially, suitable liquid solvents are: aromatics such as xylene, toluene or alkylnaphthalenes, chlorinated aromatics or chlorinated aliphatic hydrocarbons such as chlorobenzenes, chloroethylenes or methylene chloride, aliphatic hydrocarbons such as cyclohexane or paraffins, for example petroleum fractions, mineral and vegetable oils, alcohols such as butanol or glycol and also their ethers and esters, ketones such as acetone, methyl ethyl ketone, methyl isobutyl ketone or cyclohexanone, strongly polar solvents such as dimethylformamide and dimethyl sulphoxide, and also water.","As solid carriers there are suitable: for example ammonium salts and ground natural minerals such as kaolins, clays, talc, chalk, quartz, attapulgite, montmorillonite or diatomaceous earth, and ground synthetic minerals, such as highly disperse silica, alumina and silicates; as solid carriers for granules there are suitable: for example crushed and fractionated natural rocks such as calcite, marble, pumice, sepiolite and dolomite, and also synthetic granules of inorganic and organic meals, and granules of organic material such as sawdust, coconut shells, maize cobs and tobacco stalks; as emulsifiers and/or foam-formers there are suitable: for example nonionic and anionic emulsifiers, such as polyoxyethylene fatty acid esters, polyoxyethylene fatty alcohol ethers, for example alkylaryl polyglycol ethers, alkylsulphonates, alkyl sulphates, arylsulphonates and also protein hydrolysates; as dispersants there are suitable: for example lignin-sulphite waste liquors and methylcellulose.","Tackifiers such as carbox),methylcellulose and natural and synthetic polymers in the form of powders, granules or latices, such as gum arabic, polyvinyl alcohol and polyvinyl acetate, as well as natural phospholipids such as cephalins and lecithins, and synthetic phospholipids, can be used in the formulations.","Other possible additives are mineral and vegetable oils.","It is possible to use colorants such as inorganic pigments, for example iron oxide, titanium oxide and Prussian Blue, and organic dyestuffs, such as alizarin dyestuffs, azo dyestuffs and metal phthalocyanine dyestuffs, and trace nutrients such as salts of iron, manganese, boron, copper, cobalt, molybdenum and zinc.","The formulations generally comprise between 0.1 and 95% by weight of active compound, preferably between 0.5 and 90%.","The active compounds according to the invention can be employed formulations in their as such or as a mixture with known fungicides, bactericides, acaricides, nematicides or insecticides in order to increase the activity spectrum or avoid the development of resistance.","In many cases synergistic effects are achieved, ie.","the efficacy of the mixture is greater than the efficacy of the individual components.","Favourable examples of co-components in mixtures are the following compounds: Fungicides: aldimorph, ampropylfos, ampropylfos-potassium, andoprim, anilazine, azaconazole, azoxystrobin, benalaxyl, benodanil, benomyl, benzamacril, benzamacryl-isobutyl, bialaphos, binapacryl, biphenyl, bitertanoi, blasticidin-S, bromuconazole, bupirimate, buthiobate, calcium polysulphide, capsimycin, captafol, captan, carbendazim, carboxin, carvon, quinomethionate, chlobenthiazone, chlorfenazole, chloroneb, chloropicrin, chlorothalonil, chlozolinate, clozylacon, cufraneb, cymoxanil, cyproconazole, cyprodinil, cyprofuram, debacarb, dichlorophen, diclobutrazole, diclofluanid, diclomezine, dicloran, diethofencarb, difenoconazole, dimethirimol, dimethomorph, diniconazole, diniconazole-M, dinocap, diphenylamine, dipyrithione, ditalimfos, dithianon, dodemorph, dodine, drazoxolon, edifenphos, epoxiconazole, etaconazole, ethirimol, etridiazole, famoxadon, fenapanil, fenarimol, fenbuconazole, fenfuram, fenitropan, fenpiclonil, fenpropidin, fenpropimorph, fentin acetate, fentin hydroxide, ferbam, ferimzone, fluazinam, flumetover, fluoromide, fluquinconazole, flurprimidol, flusilazole, flusulfamide, flutolanil, flutriafol, folpet, fosetyl-aluminium, fosetyl-sodium, fthalide, fuberidazole, furalaxyl, furametpyr, furcarbonil, furconazole, furconazole-cis, furmecyclox, guazatine, hexachlorobenzene, hexaconazole, hymexazole, imazalil, imibenconazole, iminoctadine, iminoctadine albesilate, iminoctadine triacetate, iodocarb, ipconazole, iprobenfos (IBP), iprodione, irumamycin, isoprothiolane, isovaledione, kasugamycin, kresoxim-methyl, copper preparations, such as: copper hydroxide, copper naphthenate, copper oxychloride, copper sulphate, copper oxide, oxine-copper and Bordeaux mixture, mancopper, mancozeb, maneb, meferimzone, mepanipyrim, mepronil, metalaxyl, metconazole, methasulfocarb, methfuroxam, metiram, metomeclam, metsulfovax, mildiomycin, myclobutanil, myclozolin, nickel dimethyldithiocarbamate, nitrothal-isopropyl, nuarimol, ofurace, oxadixyl, oxamocarb, oxolinic acid, oxycarboxim, oxyfenthiin, paclobutrazole, pefurazoate, penconazole, pencycuron, phosdiphen, picoxystrobin, pimaricin, piperalin, polyoxin, polyoxorim, probenazole, prochloraz, procymidone, propamocarb, propanosine-sodium, propiconazole, propineb, pyraclostrobin, pyrazophos, pyrifenox, pyrimethanil, pyroquilon, pyroxyfur, quinconazole, quintozene (PCNB), sulphur and sulphur preparations, tebuconazole, tecloftalam, tecnazene, tetcyclacis, tetraconazole, thiabendazole, thicyofen, thifluzamide, thiophanate-methyl, thiram, tioxyrmid, tolclofos-methyl, tolyfluanid, triadimefon, triadimenol, triazbutil, triazoxide, trichlamide, tricyclazole, tridemorph, trifloxystrobin, triflurnizole, triforine, triticonazole, uniconazole, validamycin A, vinclozolin, viniconazole, zarilamide, zineb, ziram and also Dagger G, OK-8705, OK-8801, α-(1,1-dimethylethyl)-β-(2-phenoxyethyl)-1H-1,2,4-triazole-1-ethanol, α-(2,4-dichlorophenyl)-β-fluoro-β-propyl-1H-1,2,4-triazole-1-ethanol, α-(2,4-dichlorophenyl)-β-methoxy-α-methyl-1H-1,2,4-triazole-1-ethanol, α-(5-methyl-1,3-dioxan-5-yl)-β-[[4-(trifluoromethyl)-phenyl]-methylene]-1H-1,2,4-triazole-1-ethanol, (5RS,6RS)-6-hydroxy-2,2,7,7-tetramethyl-5-(1H-1,2,4-triazol-1-yl)-3-octanone, (E)-α-(methoxyimino)-N-methyl-2-phenoxy-phenylacetamide, 1-isopropyl {2-methyl-1-[[[1-(4-methylphenyl)-ethyl]-amino]-carbonyl]-propyl}-carbamate, 1-(2,4-dichlorophenyl)-2-(1H-1,2,4-triazol-1-yl)-ethanone-O-(phenylmethyl)-oxime, 1-(2-methyl-1-naphthalenyl)-1H-pyrrol-2,5-dione, 1-(3,5-dichlorophenyl)-3-(2-propenyl)-2,5-pyrrolidindione, 1-[(diiodomethyl)-sulphonyl]4-methyl-benzene, 1-[[2-(2,4-dichlorophenyl)-1,3-dioxolan-2-yl]-methyl]-1H-imidazole, 1-[[2-(4-chlorophenyl)-3-phenyloxiranyl]-methyl]-1H-1,2,4-triazole, 1-[1-[2-[(2,4-dichlorophenyl)-methoxy]-phenyl]-ethenyl]-1H-imidazole, 1-methyl-5-nonyl-2-(phenylmethyl)-3-pyrrolidinole, 2′,6′-dibromo-2-methyl4′-trifluoromethoxy4′-trifluoro-methyl-1,3-thiazole-5-carboxanilide, 2,2-dichloro-N-[1-(4-chlorophenyl)-ethyl]-1-ethyl-3-methyl-cyclopropanecarboxamide, 2,6-dichloro-5-(methylthio)4-pyrimidinyl-thiocyanate, 2,6-dichloro-N-(4-trifluoromethylbenzyl)-benzamide, 2,6-dichloro-N-[[4-(trifluoromethyl)-phenyl]-methyl]-benzamide, 2-(2,3,3-triiodo-2-propenyl)-2H-tetrazole, 2-[(1-methylethyl)-sulphonyl]-5-(trichloromethyl)-1,3,4-thiadiazole, 2-[[6-deoxy4-O-(4-O-methyl-β-D-glycopyranosyl)-α-D-glucopyranosyl]-amino]-4-methoxy-1H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile, 2-aminobutane, 2-bromo-2-(bromomethyl)-pentanedinitrile, 2-chloro-N-(2,3-dihydro-1,1,3-trimethyl-1H-inden4yl)-3-pyridinecarboxamide, 2-chloro-N-(2,6-dimethylphenyl)-N-(isothiocyanatomethyl)-acetamide, 2-phenylphenol (OPP), 3,4-dichloro-1-[4-(difluoromethoxy)-phenyl]-1H-pyrrol-2,5-dione, 3,5-dichloro-N-[cyano[(1-methyl-2-propynyl)-oxy]-methyl]-benzamide, 3-(1,1-dimethylpropyl-1-oxo-1H-indene-2-carbonitrile, 3-[2-(4-chlorophenyl)-5-ethoxy-3-isoxazolidinyl]-pyridine, 4-chloro-2-cyano-N,N-dimethyl-5-(4-methylphenyl)-1H-imidazole-1-sulphonamide, 4-methyl-tetrazolo[1,5-a]quinazolin-5(4H)-one, 8-(1,1-dimethylethyl)-N-ethyl-N-propyl-1,4-dioxaspiro[4.5]decane-2-methanamine, 8-hydroxyquinoline sulphate, 9H-xanthene-2-[(phenylamino)-carbonyl]-9-carboxylic hydrazide, bis-(1-methylethyl)-3-methyl4-[(3-methylbenzoyl)-oxy]-2,5-thiophenedicarboxylate, cis-1-(4-chlorophenyl)-2-(1H-1,2,4-triazol-1-yl)-cycloheptanol, cis4-[3-[4-(1,1-dimethylpropyl)-phenyl-2-methylpropyl]-2,6-dimethyl-morpholine hydrochloride, ethyl [(4-chlorophenyl)-azo]-cyanoacetate, potassium bicarbonate, methanetetrathiol-sodium salt, methyl 1-(2,3-dihydro-2,2-dimethyl-1H-inden-1-yl)-1H-imidazole-5-carboxylate, methyl N-(2,6-dimethylphenyl)-N-(5-isoxazolylcarbonyl)-DL-alaninate, methyl N-(chloroacetyl)-N-(2,6-dimethylphenyl)-DL-alaninate, N-(2,3-dichloro-4-hydroxyphenyl)-1-methyl-cyclohexanecarboxamide, N-(2,6-dimethylphenyl)-2-methoxy-N-(tetrahydro-2-oxo-3-furanyl)-acetamide, N-(2,6-dimethylphenyl)-2-methoxy-N-(tetrahydro-2-oxo-3-thienyl)-acetamide, N-(2-chloro-4-nitrophenyl)-4-methyl-3-nitro-benzenesulphonamide, N-(4-cyclohexylphenyl)-1,4,5,6-tetrahydro-2-pyrimidinamine, N-(4-hexylphenyl)-1,4,5,6-tetrahydro-2-pyrimidinamine, N-(5-chloro-2-methylphenyl)-2-methoxy-N-(2-oxo-3-oxazolidinyl)-acetamide, N-(6-methoxy)-3-pyridinyl-cyclopropanecarboxamide, N-[2,2,2-trichloro-1-[(chloroacetyl)-amino]-ethyl]-benzamide, N-[3-chloro-4,5-bis(2-propinyloxy)-phenyl]-N′-methoxy-methanimidamide, N-formyl-N-hydroxy-DL-alanine-sodium salt, O,O-diethyl [2-(dipropylamino)-2-oxoethyl]-ethylphosphoramidothioate,.","O-methyl S-phenyl phenylpropylphosphoramidothioate, S-methyl 1,2,3-benzothiadiazole-7-carbothioate, spiro[2H]-1-benzopyrane-2,1′(3′H)-isobenzofuran]-3′-one, 4-[(3,4-dimethoxyphenyl)-3-(4-fluorophenyl)-acryloyl]-morpholine.","Bactericides: bronopol, dichlorophen, nitrapyrin, nickel dimethyldithiocarbamate, kasugamycin, octhilinone, furancarboxylic acid, oxytetracyclin, probenazole, streptomycin, tecloftalam, copper sulphate and other copper preparations.","Insecticides/Acaricides/Nematicides: abamectin, acephate, acetamiprid, acrinathrin, alanycarb, aldicarb, aldoxycarb, alpha-cypermethrin, alphamethrin, amitraz, avermectin, AZ 60541, azadirachtin, azamethiphos, azinphos A, azinphos M, azocyclotin, Bacillus popilliae, Bacillus sphaericus, Bacillus subtilis, Bacillus thuringiensis, baculoviruses, Beauveria bassiana, Beauveria tenella, bendiocarb, benfuracarb, bensultap, benzoximate, betacyfluthrin, bifenazate, bifenthrin, bioethanomethrin, biopermethrin, bistrifluron, BPMC, bromophos A, bufencarb, buprofezin, butathiofos, butocarboxim, butylpyridaben, cadusafos, carbaryl, carbofuran, carbophenothion, carbosulfan, cartap, chloethocarb, chlorethoxyfos, chlorfenapyr, chlorfenvinphos,- chlorfluazuron, chlormephos, chlorpyrifos, chlorpyrifos M, chlovaporthrin, chromafenozide, cis-resmethrin, cispertmethrin, clocythrin, cloethocarb, clofentezine, clothianidine, cyanophos, cycloprene, cycloprothrin, cyfluthrin, cyhalothrin, cyhexatin, cypermethrin, cyromazine, deltamethrin, demeton M, demeton S, demeton-S-methyl, diafenthiuron, diazinon, dichlorvos, diflubenzuron, dimethoate, dimethylvinphos, diofenolan, disulfoton, docusat-sodium, dofenapyn, eflusilanate, emamectin, empenthrin, endosulfan, Entomopfthora spp., esfenvalerate, ethiofencarb, ethion, ethoprophos, etofenprox, etoxazole, etrimfos, fenamiphos, fenazaquin, fenbutatin oxide, fenitrothion, fenothiocarb, fenoxacrim, fenoxycarb, fenpropathrin, fenpyrad, fenpyrithrin, fenpyroximate, fenvalerate, fipronil, fluazinam, fluazuron, flubrocythrinate, flucycloxuron, flucythrinate, flufenoxuron, flumethrin, flutenzine, fluvalinate, fonophos, fosmethilan, fosthiazate, fubfenprox, furathiocarb, granulosis viruses, halofenozide, HCH, heptenophos, hexaflumuron, hexythiazox, hydroprene, imidacloprid, indoxacarb, isazofos, isofenphos, isoxathion, ivermectin, nuclear polyhedrosis viruses, lambda-cyhalothrin, lufenuron, malathion, mecarbam, metaldehyde, methamidophos, Metharhizium anisopliae, metharhizium flavoviride, methidathion, methiocarb, methoprene, methomyl, methoxyfenozide, metolcarb, metoxadiazone, mevinphos, milbemectin, milbemycin, monocrotophos, naled, nitenpyram, nithiazine, novaluron, omethoate, oxamyl, oxydemethon M, Paecilomyces fumosoroseus, parathion A, parathion M, permethrin, phenthoate, phorate, phosalone, phosmet, phosphamidon, phoxim, pirimicarb, pirimiphos A, pirimiphos M, profenofos, promecarb, propargite, propoxur, prothiofos, prothoate, pymetrozine, pyraclofos, pyresmethrin, pyrethrum, pyridaben, pyridathion, pyrimidifen, pyriproxyfen, quinalphos, ribavirin, salithion, sebufos, silafluofen, spinosad, spirodiclofen, sulfotep, sulprofos, tau-fluvalinate, tebufenozide, tebufenpyrad, tebupirimiphos, teflubenzuron, tefluthrin, temephos, temivinphos, terbufos, tetrachlorvinphos, tetradifon, theta-cypermethrin, thiacloprid, thiamethoxam, thiapronil, thiatriphos, thiocyclam hydrogen oxalate, thiodicarb, thiofanox, thuringiensin, tralocythrin, tralomethrin, triarathene, triazamate, triazophos, triazuron, trichlophenidine, trichlorfon, triflumuron, trimethacarb, vamidothion, vaniliprole, Verticillium lecanii, YI 5302, zeta-cypermethrin, zolaprofos, (1R-cis)-[5-(phenylmethyl)-3-furanyl]-methyl-3-[(dihydro-2-oxo-3(2H)-furanylidene)-methyl]-2,2-dimethylcyclopropanecarboxylate, (3-phenoxyphenyl)-methyl-2,2,3,3-tetramethylcyclopropanecarboxylate, 1-[(2-chloro-5-thiazolyl)methyl]tetrahydro-3,5-dimethyl-N-nitro-1,3,5-triazine-2(1H)-imine, 2-(2-chloro-6-fluorophenyl)4-[4-(1,1-dimethylethyl)phenyl]-4,5-dihydro-oxazole, 2-(acetyloxy)-3-dodecyl-1,4-naphthalenedione, 2-chloro-N-[[[4-(1-phenylethoxy)-phenyl]-amino]-carbonyl]-benzamide, 2-chloro-N-[[[4-(2,2-dichloro-1,1-difluoroethoxy)-phenyl]-amino]-carbonyl]-benzamide, 3-methylphenyl propylcarbamate, 4-[4-(4-ethoxyphenyl)-4-methylpentyl]-1-fluoro-2-phenoxy-benzene, 4-chloro-2-(1,1-dimethylethyl)-5-[[2-(2,6-dimethyl-4-phenoxyphenoxy)ethyl]thio]-3(2H)-pyridazinone, 4-chloro-2-(2-chloro-2-methylpropyl)-5-[(6-iodo-3-pyridinyl)methoxy]-3(2H)-pyridazinone, 4-chloro-5-[(6-chloro-3-pyridinyl)methoxy]-2-(3,4-dichlorophenyl)-3(2H)-pyridazinone, Bacillus thuringiensis strain EG-2348, [2-benzoyl-1-(1,1-dimethylethyl)-hydrazinobenzoic acid, 2,2-dimethyl-3-(2,4-dichlorophenyl)-2-oxo-1-oxaspiro[4.5]dec-3-en4-yl butanoate, [3-[(6-chloro-3-pyridinyl)methyl]-2-thiazolidinylidene]-cyanamide, dihydro-2-(nitromethylene)-2H-1,3-thiazine-3(4H)-carboxaldehyde, ethyl [2-[[1,6-dihydro-6-oxo-1-(phenylmethyl)-4-pyridazinyl]oxy]ethyl]-carbamate, N-(3,4,4-trifluoro-1-oxo-3-butenyl)-glycine, N-(4-chlorophenyl)-3-[4-(difluoromethoxy)phenyl]4,5-dihydro4-phenyl-1H-pyrazole-1-carboxamide, N-[(2-chloro-5-thiazolyl)methyl]-N′methyl-N″-nitro-guanidine, N-methyl-N′-(1-methyl-2-propenyl)-1,2-hydrazinedicarbothioamide, N-methyl-N′-2-propenyl-1,2-hydrazinedicarbothioamide, O,O-diethyl-[2-(dipropylamino)-2-oxoethyl]-ethylphosphoramidothioate, N-cyanomethyl-4-trifluoromethyl-nicotinamide, 3,5-dichloro-1-(3,3-dichloro-2-propenyloxy)-4-[3-(5-trifluoromethylpyridin-2-yloxy)-propoxy]-benzene.","It is also possible to admix other known active compounds, such as herbicides, fertilizers and growth regulators.","When used as insecticides, the active compounds according to the invention can furthermore be present in their commercially available formulations and in the use forms, prepared from these formulations, as a mixture with synergistic agents.","Synergistic agents are compounds which increase the action of the active compounds, without it being necessary for the synergistic agent added to be active itself.","The active compound content of the use forms prepared from the commercially available formulations can vary within wide limits.","The active compound concentration of the use forms can be from 0.0000001 to 95% by weight of active compound, preferably between 0.0001 and 1% by weight.","The compounds are employed in a customary manner appropriate for the use forms.","When used against hygiene pests and pests of stored products, the active compound is distinguished by an excellent residual action on wood and clay as well as by a good stability to alkali on limed substrates.","As already mentioned above, it is possible to treat all plants and their parts according to the invention.","In a preferred embodiment, wild plant species and plant cultivars, or those obtained by conventional biological breeding, such as crossing or protoplast fusion, and parts thereof, are treated.","In a further preferred embodiment, transgenic plants and plant cultivars obtained by genetic engineering, if appropriate in combination with conventional methods (Genetically Modified Organisms), and parts thereof are treated.","The term “parts” or “parts of plants” or “plant parts” has been explained above.","Particularly preferably, plants of the plant cultivars which are in each case commercially available or in use are treated according to the invention.","Plant cultivars are understood as meaning plants with novel properties (“traits”) which are grown by conventional cultivation, by mutagenesis or by recombinant DNA techniques.","These may be cultivars, biotypes or genotypes.","Depending on the plant species or plant cultivars, their location and growth conditions (soils, climate, vegetation period, diet), the treatment according to the invention may also result in superadditive (“synergistic”) effects.","Thus, for example, reduced application rates and/or a widening of the activity spectrum and/or an increase in the activity of the substances and compositions to be used according to the invention, better plant growth, increased tolerance to high or low temperatures, increased tolerance to drought or to water or soil salt content, increased flowering performance, easier harvesting, accelerated maturation, higher harvest yields, better quality and/or a higher nutritional value of the harvested products, better storage stability and/or processability of the harvested products are possible which exceed the effects which were actually to be expected.","The transgenic plants or plant cultivars (i.e.","those obtained by genetic engineering) which are preferably to be treated according to the invention include all plants which, in the genetic modification, received genetic material which imparted particularly advantageous useful properties (“traits”) to these plants.","Examples of such properties are better plant growth, increased tolerance to high or low temperatures, increased tolerance to drought or to water or soil salt content, increased flowering performance, easier harvesting, accelerated maturation, higher harvest yields, better quality and/or a higher nutritional value of the harvested products, better storage stability and/or processability of the harvested products.","Further and particularly emphasized examples of such properties are a better defence of the plants against animal and microbial pests, such as against insects, mites, phytopathogenic fungi, bacteria and/or viruses, and also increased tolerance of the plants to certain herbicidally active compounds.","Examples of transgenic plants which may be mentioned are the important crop plants, such as cereals (wheat, rice), maize, soya beans, potatoes, cotton, oilseed rape and also fruit plants (with the fruits apples, pears, citrus fruits and grapes), and particular emphasis is given to maize, soya beans, potatoes, cotton and oilseed rape.","Traits that are emphasized are in particular increased defence of the plants against insects by toxins formed in the plants, in particular those formed in the plants by the genetic material from Bacillus thuringiensis (for example by the genes CryIA(a), CryIA(b), CryIA(c), CryIIA, CryIIIA, CryIIIB2, Cry9c Cry2Ab, Cry3Bb and CryIF and also combinations thereof) (hereinbelow referred to as “Bt plants”).","Traits which are also particularly emphasized are the increased resistance of plants to fungi, bacteria and viruses by systemic acquired resistance (SAR), systemin, phytoalexins, elicitors and resistance genes and the correspondingly expressed proteins and toxins.","Traits that are furthermore particularly emphasized are the increased tolerance of the plants to certain herbicidally active compounds, for example imidazolinones, sulphonylureas, glyphosate or phosphinotricin (for example the “PAT” gene).","The genes which impart the desired traits in question can also be present in combination with one another in the transgenic plants.","Examples of “Bt plants” which may be mentioned are maize varieties, cotton varieties, soya bean varieties and potato varieties which are sold under the trade names YIELD GARD® (for example maize, cotton, soya beans), KnockOut® (for example maize), StarLink® (for example maize), Bollgard® (cotton), Nucotn® (cotton) and NewLeaf® (potato).","Examples of herbicide-tolerant plants which may be mentioned are maize varieties, cotton varieties and soya bean varieties which are sold under the trade names Roundup Ready® (tolerance to glyphosate, for example maize, cotton, soya bean), Liberty Link® (tolerance to phosphinotricin, for example oilseed rape), IMI® (tolerance to imidazolinones) and STS® (tolerance to sulphonylureas, for example maize).","Herbicide-resistant plants (plants bred in a conventional manner for herbicide tolerance) which may be mentioned include the varieties sold under the name Clearfield® (for example maize).","Of course, these statements also apply to plant cultivars having these genetic traits or genetic traits still to be developed, which cultivars will be developed and/or marketed in the future.","The plants listed can be treated according to the invention in a particularly advantageous manner with the compounds of the formula I or the active compound mixtures according to the invention.","The preferred ranges stated above for the active compounds or mixtures also apply to the treatment of these plants.","Particular emphasis is given to the treatment of plants with the compounds or mixtures specifically mentioned in the present text.","The active compounds according to the invention act not only against plant, hygiene and stored product pests, but also in the veterinary medicine sector against animal parasites (ectoparasites), such as hard ticks, soft ticks, mange mites, leaf mites, flies (biting and licking), parasitic fly larvae, lice, hair lice, feather lice and fleas.","These parasites include: From the order of the Anoplurida, for example, Haematopinus spp., Linognathus spp., Pediculus spp., Phtirus spp.","and Solenopotes spp.","From the order of the Mallophagida and the suborders Amblycerina and Ischnocerina, for example, Trimenopon spp., Menopon spp., Trinoton spp., Bovicola spp., Werneckiella spp., Lepikentron spp., Damalina spp., Trichodectes spp.","and Felicola spp.","From the order of the Diptera and the suborders Nematocerina and Brachycerina, for example, Aedes spp., Anopheles spp., Culex spp., Simulium spp., Eusimulium spp., Phlebotomus spp., Lutzomyia spp., Culicoides spp., Chrysops spp., Hybomitra spp., Atylotus spp., Tabanus spp., Haematopota spp., Philipomyia spp., Braula spp., Musca spp., Hydrotaea spp., Stomoxys spp., Haematobia spp., Morellia spp., Fannia spp., Glossina spp., Calliphora spp., Lucilia spp., Chrysomyia spp., Wohlfahrtia spp., Sarcophaga spp., Oestrus spp., Hypoderma spp., Gasterophilus spp., Hippobosca spp., Lipoptena spp.","and Melophagus spp.","From the order of the Siphonapterida, for example Pulex spp., Ctenocephalides spp., Xenopsylla spp.","and Ceratophyllus spp.","From the order of the Heteropterida, for example, Cimex spp., Triatoma spp., Rhodnius spp.","and Panstrongylus spp.","From the order of the Blattarida, for example Blatta orientalis, Periplaneta americana, Blattela germanica and Supella spp.","From the subclass of the Acaria (Acarida) and the orders of the Meta- and Mesostigmata, for example, Argas spp., Ornithodorus spp., Otobius spp., Ixodes spp., Amblyomma spp., Boophilus spp., Dermacentor spp., Haemophysalis spp., Hyalomma spp., Rhipicephalus spp., Dermanyssus spp., Raillietia spp., Pneumonyssus spp., Sternostoma spp.","and Varroa spp.","From the order of the Actinedida (Prostigmata) und Acaridida (Astigmata), for example, Acarapis spp., Cheyletiella spp., Ornithocheyletia spp., Myobia spp., Psorergates spp., Demodex spp., Trombicula spp., Listrophorus spp., Acarus spp., Tyrophagus spp., Caloglyphus spp., Hypodectes spp., Pterolichus spp., Psoroptes spp., Chorioptes spp., Otodectes spp., Sarcoptes spp., Notoedres spp., Knemidocoptes spp., Cytodites spp.","and Laminosioptes spp.","The active compounds of the formula (I) according to the invention are also suitable for controlling arthropods which infest agricultural productive livestock, such as, for example, cattle, sheep, goats, horses, pigs, donkeys, camels, buffalo, rabbits, chickens, turkeys, ducks, geese and bees, other pets, such as, for example, dogs, cats, caged birds and aquarium fish, and also so-called test animals, such as, for example, hamsters, guinea pigs, rats and mice.","By controlling these arthropods, cases of death and reduction in productivity (for meat, milk, wool, hides, eggs, honey etc.)","should be diminished, so that more economic and easier animal husbandry is possible by use of the active compounds according to the invention.","The active compounds according to the invention are used in the veterinary sector in a known manner by enteral administration in the form of, for example, tablets, capsules, potions, drenches, granules, pastes, boluses, the feed-through process and suppositories, by parenteral administration, such as, for example, by injection (intramuscular, subcutaneous, intravenous, intraperitoneal and the like), implants, by nasal administration, by dermal use in the form, for example, of dipping or bathing, spraying, pouring on and spotting on, washing and powdering, and also with the aid of molded articles containing the active compound, such as collars, ear marks, tail marks, limb bands, halters, marking devices and the like.","When used for cattle, poultry, pets and the like, the active compounds of the formula (I) can be used as formnulations (for example powders, emulsions, free-flowing compositions), which comprise the active compounds in an amount of 1 to 80% by weight, directly or after 100 to 10 000-fold dilution, or they can be used as a chemical bath.","It has furthermore been found that the compounds according to the invention have a strong insecticidal action against insects which destroy industrial materials.","The following insects may be mentioned as examples and as preferred—but without a limitation: Beetles, such as Hylotrupes bajulus, Chlorophorus pilosis, Anobium punctatum, Xestobium rufovillosum, Ptilinus pecticomis, Dendrobium pertinex, Ernobius mollis, Priobium carpini, Lyctus brunneus, Lyctus africanus, Lyctus planicollis, Lyctus linearis, Lyctus pubescens, Trogoxylon aequale, Minthes rugicollis, Xyleborus spec.","Tryptodendron spec.","Apate monachus, Bostrychus capucins, Heterobostrychus brunneus, Sinoxylon spec.","Dinoderus minutus.","Hymenopterons, such as Sirex juvencus, Urocerus gigas, Urocerus gigas taignus, Urocerus augur.","Termites, such as Kalotermes flavicollis, Cryptotermes brevis, Heterotermes indicola, Reticulitermes flavipes, Reticulitermes santonensis, Reticulitermes lucifugus, Mastotermes darwiniensis, Zootermopsis nevadensis, Coptotermes formosanus.","Bristletails, such as Lepisma saccharina.","Industrial materials in the present connection are to be understood as meaning non-living materials, such as, preferably, plastics, adhesives, sizes, papers and cards, leather, wood and processed wood products and coating compositions.","Wood and processed wood products are materials to be protected, especially preferably, from insect infestation.","Wood and processed wood products which can be protected by the agents according to the invention or mixtures comprising these are to be understood as meaning, for example: building timber, wooden beams, railway sleepers, bridge components, boat jetties, wooden vehicles, boxes, pallets, containers, telegraph poles, wood panelling, wooden windows and doors, plywood, chipboard, joinery or wooden products which are used quite generally in house-building or in building joinery.","The active compounds can be used as such; in the form of concentrates or generally customary formulations, such as powders, granules, solutions, suspensions, emulsions or pastes.","The formulations mentioned can be prepared in a manner known per se, for example by mixing the active compounds with at least one solvent or diluent, emulsifier, dispersing agent and/or binder or fixing agent, a water repellent, if appropriate siccatives and UV stabilizers and if appropriate dyestuffs and pigments, and also other processing auxiliaries.","The insecticidal compositions or concentrates used for the preservation of wood and wood-derived timber products comprise the active compound according to the invention in a concentration of 0.0001 to 95% by weight, in particular 0.001 to 60% by weight.","The amount of the compositions or concentrates employed depends on the nature and occurrence of the insects and on the medium.","The optimum amount employed can be determined for the use in each case by a series of tests.","In general, however, it is sufficient to employ 0.0001 to 20% by weight, preferably 0.001 to 10% by weight, of the active compound, based on the material to be preserved.","Solvents and/or diluents which are used are an organic chemical solvent or solvent mixture and/or an oily or oil-like organic chemical solvent or solvent mixture of low volatility and/or a polar organic chemical solvent or solvent mixture and/or water, and if appropriate an emulsifier and/or wetting agent.","Organic chemical solvents which are preferably used are oily or oil-like solvents having an evaporation number above 35 and a flashpoint above 30° C., preferably above 45° C. Substances which are used as such oily or oil-like water-insoluble solvents of low volatility are appropriate mineral oils or aromatic fractions thereof, or solvent mixtures containing mineral oils, preferably white spirit, petroleum and/or alkylbenzene.","Mineral oils having a boiling range from 170 to 220° C., white spirit having a boiling range from 170 to 220° C., spindle oil having a boiling range from 250 to 350° C., petroleum and aromatics having a boiling range from 160 to 280° C., terpentine oil and the like, are advantageously employed.","In a preferred embodiment, liquid aliphatic hydrocarbons having a boiling range from 180 to 210° C. or high-boiling mixtures of aromatic and aliphatic hydrocarbons having a boiling range from 180 to 220° C. and/or spindle oil and/or monochloronaphthalene, preferably α-monochloronaphthalene, are used.","The organic oily or oil-like solvents of low volatility which have an evaporation number above 35 and a flashpoint above 30° C., preferably above 45° C., can be replaced in part by organic chemical solvents of high or medium volatility, providing that the solvent mixture likewise has an evaporation number above 35 and a flashpoint above 30° C., preferably above 45° C., and that the insecticide/fungicide mixture is soluble or emulsifiable in this solvent mixture.","According to a preferred embodiment, some of the organic chemical solvent or solvent mixture or an aliphatic polar organic chemical solvent or solvent mixture is replaced.","Aliphatic organic chemical solvents containing hydroxyl and/or ester and/or ether groups, such as, for example, glycol ethers, esters or the like, are preferably used.","Organic chemical binders which are used in the context of the present invention are the synthetic resins and/or binding drying oils which are known per se, are water-dilutable and/or are soluble or dispersible or emulsifiable in the organic chemical solvents employed, in particular binders consisting of or comprising an acrylate resin, a vinyl resin, for example polyvinyl acetate, polyester resin, polycondensation or polyaddition resin, polyurethane resin, alkyd resin or modified alkyd resin, phenolic resin, hydrocarbon resin, such as indene-cumarone resin, silicone resin, drying vegetable oils and/or drying oils and/or physically drying binders based on a natural and/or synthetic resin.","The synthetic resin used as the binder can be employed in the form of an emulsion, dispersion or solution.","Bitumen or bituminous substances can also be used as binders in an amount of up to 10% by weight.","Dyestuffs, pigments, water-repelling.","agents, odour correctants and inhibitors or anticorrosive agents and the like which are known per se can additionally be employed.","It is preferred according to the invention for the composition or concentrate to comprise, as the organic chemical binder, at least one alkyd resin or modified alkyd resin and/or one drying vegetable oil.","Alkyd resins having an oil content of more than 45% by weight, preferably 50 to 68% by weight, are preferably used according to the invention.","All or some of the binder mentioned can be replaced by a fixing agent (mixture) or a plasticizer (mixture).","These additives are intended to prevent evaporation of the active compounds and crystallization or precipitation.","They preferably replace 0.01 to 30% of the binder (based on 100% of the binder employed).","The plasticizers originate from the chemical classes of phthalic acid esters, such as dibutyl, dioctyl or benzyl butyl phthalate, phosphoric acid esters, such as tributyl phosphate, adipic acid esters, such as di-(2-ethylhexyl) adipate, stearates, such as butyl stearate or amyl stearate, oleates, such as butyl oleate, glycerol ethers or higher molecular weight glycol ethers, glycerol esters and p-toluenesulphonic acid esters.","Fixing agents are based chemically on polyvinyl alkyl ethers, such as, for example, polyvinyl methyl ether or ketones, such as benzophenone or ethylenebenzophenone.","Possible solvents or diluents are, in particular, also water, if appropriate as a mixture with one or more of the abovementioned organic chemical solvents or diluents, emulsifiers and dispersing agents.","Particularly effective preservation of wood is achieved by impregnation processes on a large industrial scale, for example vacuum, double vacuum or pressure processes.","The, ready-to-use compositions can also comprise other insecticides, if appropriate, and also one or more fungicides, if appropriate.","Possible additional mixing partners are, preferably, the insecticides and fungicides mentioned in WO 94/29 268.The compounds mentioned in this document are an explicit constituent of the present application.","Especially preferred mixing partners which may be mentioned are insecticides, such as chlorpyriphos, phoxim, silafluofin, alphamethrin, cyfluthrin, cypermethrin, deltamethrin, permethrin, imidacloprid, NI-25, flufenoxuron, hexaflumuron, transfluthrin, thiacloprid, methoxyfenozide and triflumuron, and also fungicides, such as epoxyconazole, hexaconazole, azaconazole, propiconazole, tebuconazole, cyproconazole, metconazole, imazalil, dichlorfluanid, tolylfluanid, 3-iodo-2-propinyl-butyl carbamate, N-octyl-isothiazolin-3-one and 4,5-dichloro-N-octylisothiazolin-3-one.","The compounds according to the invention can at the same time be employed for protecting objects which come into contact with saltwater or brackish water, such as hulls, screens, nets, buildings, moorings and signalling systems, against fouling.","Fouling by sessile Oligochaeta, such as Serpulidae, and by shells and species from the Ledamorpha group (goose barnacles), such as various Lepas and Scalpellum species, or by species from the Balanomorpha group (acorn barnacles), such as Balanus or Pollicipes species, increases the frictional drag of ships and, as a consequence, leads to a marked increase in operation costs owing to higher energy consumption and additionally frequent residence in the dry dock.","Apart from fouling by algae, for example Ectocarpus sp.","and Ceramium sp., in particular fouling by sessile Entomostraka groups, which come under the generic term Cirripedia (cirriped crustaceans), is of particular importance.","Surprisingly, it has now been found that the compounds according to the invention, alone or in combination with other active compounds, have an outstanding antifouling action.","Using the compounds according to the invention, alone or in combination with other active compounds, allows the use of heavy metals such as, for example, in bis(trialkyltin) sulphides, tri-n-butyltin laurate, tri-n-butyltin chloride, copper(I) oxide, triethyltin chloride, tri-n-butyl(2-phenyl4-chlorophenoxy)tin, tributyltin oxide, molybdenum disulphide, antimony oxide, polymeric butyl titanate, phenyl-(bispyridine)bismuth chloride, tri-n-butyltin fluoride, manganese ethylenebis-thiocarbamate, zinc dimethyldithiocarbamate, zinc ethylenebisthiocarbamate, zinc salts and copper salts of 2-pyridinethiol 1-oxide, bisdimethyldithiocarbamoylzinc ethylene-bisthiocarbamate, zinc oxide, copper(I) ethylene-bisdithiocarbamate, copper thiocyanate, copper naphthenate and tributyltin halides to be dispensed with, or the concentration of these compounds to be substantially reduced.","If appropriate, the ready-to-use antifouling paints can additionally comprise other active compounds, preferably algicides, fungicides, herbicides, molluscicides, or other antifouling active compounds.","Preferably suitable components in combinations with the antifouling compositions according to the invention are: algicides such as 2-tert-butylamino-4-cyclopropylamino-6-methylthio-1,3,5-triazine, dichlorophen, diuron, endothal, fentin acetate, isoproturon, methabenzthiazuron, oxyfluorfen, quinoclamine and terbutryn; fungicides such as benzo[b]thiophenecarboxylic acid cyclohexylamide S,S-dioxide, dichlofluanid, fluorfolpet, 3-iodo-2-propinyl butylcarbamate, tolylfluanid and azoles such as azaconazole, cyproconazole, epoxyconazole, hexaconazole, metconazole, propiconazole and tebuconazole; molluscicides such as fentin acetate, metaldehyde, methiocarb, niclosamid, thiodicarb and trimethacarb; or conventional antifouling active compounds such as 4,5-dichloro-2-octyl-4-isothiazolin-3-one, diiodomethylparatryl sulphone, 2-(N,N-dimethylthiocarbamoylthio)-5-nitrothiazyl, potassium, copper, sodium and zinc salts of 2-pyridinethiol 1-oxide, pyridine-triphenylborane, tetrabutyldistannoxane, 2,3,5,6-tetrachloro-4-(methylsulphonyl)-pyridine, 2,4,5,6-tetrachloroisophthalonitrile, tetramethylthiuram disulphide and 2,4,6-trichlorophenylmaleiimide.","The antifouling compositions used comprise the active compound according to the invention of the compounds according to the invention in a concentration of 0.001 to 50% by weight, in particular 0.01 to 20% by weight.","Moreover, the antifouling compositions according to the invention comprise the customary components such as, for example, those described in Ungerer, Chem.","Ind.","1985, 37, 730-732 and Williams, Antifouling Marine Coatings, Noyes, Park Ridge, 1973.Besides the algicidal, fungicidal, molluscicidal active compounds and insecticidal active compounds according to the invention, antifouling paints comprise, in particular, binders.","Examples of recognized binders are polyvinyl chloride in a solvent system, chlorinated rubber in a solvent system, acrylic resins in a solvent system, in particular in an aqueous system, vinyl chloride/vinyl acetate copolymer systems in the form of aqueous dispersions or in the form of organic solvent systems, butadiene/styrene/acrylonitrile rubbers, drying oils such as linseed oil, resin esters or modified hardened resins in combination with tar or bitumens, asphalt and epoxy compounds, small amounts of chlorine rubber, chlorinated polypropylene and vinyl resins.","If appropriate, paints also comprise inorganic pigments, organic pigments or colorants which are preferably insoluble in salt water.","Paints may furthermore comprise materials such as colophonium to allow controlled release of the active compounds.","Furthermore, the paints may comprise plasticizers, modifiers which affect the rheological properties and other conventional constituents.","The compounds according to the invention or the abovementioned mixtures may also be incorporated into self-polishing antifouling systems.","The active compounds are also suitable for controlling animal pests, in particular insects, arachnids and mites, which are found in enclosed spaces such as, for example, dwellings, factory halls, offices, vehicle cabins and the like.","They can be employed alone or in combination with other active compounds and auxiliaries in domestic insecticide products for controlling these pests.","They are active against sensitive and resistant species and against all developmental stages.","These pests include: From the order of the Scorpionidea, for example, Buthus occitanus.","From the order of the Acarina, for example, Argas persicus, Argas reflexus, Bryobia spp., Dermanyssus gallinae, Glyciphagus domesticus, Ornithodorus moubat, Rhipicephalus sanguineus, Trombicula alfreddugesi, Neutrombicula autumnalis, Dermatophagoides pteronissimus, Dermatophagoides forinae.","From the order of the Araneae, for example, Aviculariidae, Araneidae.","From the order of the Opiliones, for example, Pseudoscorpiones chelifer, Pseudoscorpiones cheiridium, Opiliones phalangium.","From the order of the Isopoda, for example, Oniscus asellus, Porcellio scaber.","From the order of the Diplopoda, for example, Blaniulus guttulatus, Polydesmus spp.","From the order of the Chilopoda, for example, Geophilus spp.","From the order of the Zygentoma, for example, Ctenolepisma spp., Lepisma saccharina, Lepismodes inquilinus.","From the order of the Blattaria, for example, Blatta orientalies, Blattella germanica, Blattella asahinai, Leucophaea maderae, Panchlora spp., Parcoblatta spp., Periplaneta australasiae, Periplaneta americana, Periplaneta brunnea, Periplaneta fuliginosa, Supella longipalpa.","From the order of the Saltatoria, for example, Acheta domesticus.","From the order of the Dermaptera, for example, Forficula auricularia.","From the order of the Isoptera, for example, Kalotermes spp., Reticulitermes spp.","From the order of the Psocoptera, for example, Lepinatus spp., Liposcelis spp.","From the order of the Coleptera, for example, Anthrenus spp., Attagenus spp., Dermestes spp., Latheticus oryzae, Necrobia spp., Ptinus spp., Rhizopertha dominica, Sitophilus granarius, Sitophilus oryzae, Sitophilus zeamais, Stegobium paniceum.","From the order of the Diptera, for example, Aedes aegypti, Aedes albopictus, Aedes taeniorhynchus, Anopheles spp., Calliphora erythrocephala, Chrysozona pluvialis, Culex quinquefasciatus, Culex pipiens, Culex tarsalis, Drosophila spp., Fannia canicularis, Musca domestica, Phlebotomus spp., Sarcophaga carnaria, Simulium spp., Stomoxys calcitrans, Tipula paludosa.","From the order of the Lepidoptera, for example, Achroia grisella, Galleria mellonella, Plodia interpunctella, Tinea cloacella, Tinea pellionella, Tineola bisselliella.","From the order of the Siphonaptera, for example, Ctenocephalides canis, Ctenocephalides felis, Pulex irritans, Tunga penetrans, Xenopsylla cheopis.","From the order of the Hymenoptera, for example, Camponotus herculeanus, Lasius fuliginosus, Lasius niger, Lasius umbratus, Monomorium pharaonis, Paravespula spp., Tetramorium caespitum.","From the order of the Anoplura, for example, Pediculus humanus capitis, Pediculus humanus corporis, Phthirus pubis.","From the order of the Heteroptera, for example, Cimex hemipterus, Cimex lectularius, Rhodinus prolixus, Triatoma infestans.","In the field of household insecticides, they are used alone or in combination with other suitable active compounds, such as phosphoric acid esters, carbamates, pyrethroids, growth regulators or active compounds from other known classes of insecticides.","They are used as aerosols, pressure-free spray products, for example pump and atomizer sprays, automatic fogging systems, foggers, foams, gels, evaporator products with evaporator tablets made of cellulose or polymer, liquid evaporators, gel and membrane evaporators, propeller-driven evaporators, energy-free or passive, evaporation systems, moth papers, moth bags and moth gels, as granules or dusts, in baits for spreading or in bait stations.","The active compounds according to the invention can also be used as defoliants, desiccants, haulm killers and, especially, as weedkillers.","Weeds in the broadest sense are understood to mean all plants which grow in locations where they are undesired.","Whether the substances according to the invention act as total or selective herbicides depends essentially on the amount used.","The active compounds according to the invention can be used, for example, in connection with the following plants: Dicotyledonous weeds of the genera: Abutilon, Amaranthus, Ambrosia, Anoda, Anthemis, Aphanes, Atriplex, Bellis, Bidens, Capsella, Carduus, Cassia, Centaurea, Chenopodium, Cirsium, Convolvulus, Datura, Desmodium, Emex, Erysimum, Euphorbia, Galeopsis, Galinsoga, Galium, Hibiscus, Ipomoea, Kochia, Lamium, Lepidium, Lindernia, Matricaria, Mentha, Mercurialis, Mullugo, Myosotis, Papaver, Pharbitis, Plantago, Polygonum, Portulaca, Ranunculus, Raphanus, Rorippa, Rotala, Rumex, Salsola, Senecio, Sesbania, Sida, Sinapis, Solanum, Sonchus, Sphenoclea, Stellaria, Taraxacum, Thlaspi, Trifolium, Urtica, Veronica, Viola, Xanthium.","Dicotyledonous crops of the genera: Arachis, Beta, Brassica, Cucumis, Cucurbita, Helianthus, Daucus, Glycine, Gossypium, Ipomoea, Lactuca, Linum, Lycopersicon, Nicotiana, Phaseolus, Pisum, Solanum, Vicia.","Monocotyledonous weeds of the genera: Aegilops, Agropyron, Agrostis, Alopecurus, Apera, Avena, Brachiaria, Bromus, Cenchrus, Commelina, Cynodon, Cyperus, Dactyloctenium, Digitaria, Echinochloa, Eleocharis, Eleusine, Eragrostis, Eriochloa, Festuca, Fimbristylis, Heteranthera, Imperata, Ischaemum, Leptochloa, Lolium, Monochoria, Panicum, Paspalum, Phalaris, Phleum, Poa, Rottboellia, Sagittaria, Scirpus, Setaria, Sorghum.","Monocotyledonous crops of the genera: Allium, Ananas, Asparagus, Avena, Hordeum, Oryza, Panicum, Saccharum, Secale, Sorghum, Triticale, Triticum, Zea.","However, the use of the active compounds according to the invention is in no way restricted to these genera, but also extends in the same manner to other plants.","The active compounds according to the invention are suitable, depending on the concentration, for the total control of weeds, for example on industrial terrain and rail tracks, and on paths and areas with and without tree plantings.","Similarly, the active compounds according to the invention can be employed for controlling weeds in perennial crops, for example forests, decorative tree plantings, orchards, vineyards, citrus groves, nut orchards, banana plantations, coffee plantations, tea plantations, rubber plantations, oil palm plantations, cocoa plantations, soft fruit plantings and hop fields, on lawns, turf and pastureland, and for the selective control of weeds in annual crops.","The compounds of the formula (I) according to the invention have strong herbicidal activity and a broad active spectrum when used on the soil and on above-ground parts of plants.","To a certain extent they are also suitable for the selective control of monocotyledonous and dicotyledonous weeds in monocotyledonous and dicotyledonous crops, both by the pre-emergence and by the post-emergence method.","At certain concentrations or application rates, the active compounds according to the invention can also be employed for controlling animal pests and fungal or bacterial plant diseases.","If appropriate, they can also be used as intermediates or precursors for the synthesis of other active compounds.","The active compounds can be converted into the customary formulations such as solutions, emulsions, wettable powders, suspensions, powders, dusts, pastes, soluble powders, granules, suspension-emulsion concentrates, natural and synthetic materials impregnated with active compound, and microencapsulations in polymeric materials.","These formulations are produced in a known manner, for example by mixing the active compounds with extenders, that is, liquid solvents and/or solid carriers, optionally with the use of surfactants, that is, emulsifiers and/or dispersants, and/or foam formers.","If the extender used is water, it is also possible to employ, for example, organic solvents as auxiliary solvents.","Essentially, suitable liquid solvents are: aromatics such as xylene, toluene or alkylnaphthalenes, chlorinated aromatics and chlorinated aliphatic hydrocarbons such as chlorobenzenes, chloroethylenes or methylene chloride, aliphatic hydrocarbons such as cyclohexane or paraffins, for example mineral oil fractions, mineral and vegetable oils, alcohols such as butanol or glycol and their ethers and esters, ketones such as acetone, methyl ethyl ketone, methyl isobutyl ketone or cyclohexanone, strongly polar solvents such as dimethylformamide and dimethyl sulphoxide, or else water.","Suitable solid carriers are: for example ammonium salts and ground natural minerals such as kaolins, clays, talc, chalk, quartz, attapulgite, montmorillonite or diatomaceous earth, and ground synthetic materials such as finely divided silica, alumina and silicates; suitable solid carriers for granules are: for example crushed and fractionated natural rocks such as calcite, marble, pumice, sepiolite and dolomite, or else synthetic granules of inorganic and organic meals, and granules of organic material such as sawdust, coconut shells, maize cobs and tobacco stalks; suitable emulsifiers and/or foam formers are: for example nonionic and anionic emulsifiers such as polyoxyethylene fatty acid esters, polyoxyethylene fatty alcohol ethers, for example alkylaryl polyglycol ethers, alkylsulphonates, alkyl sulphates, arylsulphonates, or else protein hydrolysates; suitable dispersants are: for example lignosulphite waste liquors and methylcellulose.","Tackifiers such as carboxymethylcellulose and natural and synthetic polymers in the form of powders, granules or latices, such as gum arabic, polyvinyl alcohol and polyvinyl acetate, or else natural phospholipids such as cephalins and lecithins and synthetic phospholipids can be used in the formulations.","Other possible additives are mineral and vegetable oils.","It is possible to use colorants such as inorganic pigments, for example iron oxide, titanium oxide and Prussian Blue, and organic colorants such as alizarin colorants, azo colorants and metal phthalocyanine colorants, and trace nutrients such as salts of iron, manganese, boron, copper, cobalt, molybdenum and zinc.","The formulations generally comprise between 0.1 and 95% by weight of active compound, preferably between 0.5 and 90%.","For controlling weeds, the active compounds according to the invention, as such or in their formulations, can also be used as mixtures with known herbicides and/or substances which improve the compatibility with crop plants (“safeners”), finished formulations or tank mixes being possible.","Also possible are mixtures with weed-killers comprising one or more known herbicides and a safener.","Possible components for the mixtures are known herbicides, for example acetochlor, acifluorfen (-sodium), aclonifen, alachlor, alloxydim (-sodium), ametryne, amicarbazone, amidochlor, amidosulfuron, anilofos, asulam, atrazine, azafenidin, azimsulfuron, beflubutamid, benazolin (-ethyl), benfuresate, bensulfuron (-methyl), bentazon, benzfendizone, benzobicyclon, benzofenap, benzoylprop (-ethyl), bialaphos, bifenox, bispyribac (-sodium), bromobutide, bromofenoxim, bromoxynil, butachlor, butafenacil (-allyl), butroxydim, butylate, cafenstrole, caloxydim, carbetamide, carfentrazone (-ethyl), chlomethoxyfen, chloramben, chloridazon, chlorimuron (-ethyl), chlornitrofen, chlorsulfuron, chlortoluron, cinidon (-ethyl), cinmethylin, cinosulfuron, clefoxydim, clethodim, clodinafop (-propargyl), clomazone, clomeprop, clopyralid, clopyrasulfuron (-methyl), cloransulam (-methyl), cumyluron, cyanazine, cybutryne, cycloate, cyclosulfamuron, cycloxydim, cyhalofop (-butyl), 2,4-D, 2,4-DB, desmedipham, diallate, dicamba, dichlorprop (—P), diclofop (-methyl), diclosulam, diethatyl (-ethyl), difenzoquat, diflufenican, diflufenzopyr, dimefuron, dimepiperate, dimethachlor, dimethametryn, dimethenamid, dimexyflam, dinitramine, diphenamid, diquat, dithiopyr, diuron, dymron, epropodan, EPTC, esprocarb, ethalfluralin, ethametsulfuron (-methyl), ethofumesate, ethoxyfen, ethoxysulfuron, etobenzanid, fenoxaprop (—P-ethyl), fentrazamide, flamprop (-isopropyl, -isopropyl-L, -methyl), flazasulfuron, florasulam, fluazifop (—P-butyl), fluazolate, flucarbazone (-sodium), flufenacet, flumetsulam, flumiclorac (-pentyl), flumioxazin, flumipropyn, flumetsulam, fluometuron, fluorochloridone, fluoroglycofen (-ethyl), flupoxam, flupropacil, flurpyrsulfuron (-methyl, -sodium), flurenol (-butyl), fluridone, fluroxypyr (-butoxypropyl, -meptyl), flurprimidol, flurtamone, fluthiacet (-methyl), fluthiamide, fomesafen, foramsulfuron, glufosinate (-ammonium), glyphosate (-isopropylammonium), halosafen, haloxyfop (-ethoxyethyl, —P-methyl), hexazinone, imazamethabenz (-methyl), imazamethapyr, imazamox, imazapic, imazapyr, imazaquin, imazethapyr, imazosulfuron, iodosulfuron (-methyl, -sodium), ioxynil, isopropalin, isoproturon, isouron, isoxaben, isoxachlortole, isoxaflutole, isoxapyrifop, lactofen, lenacil, linuron, MCPA, mecoprop, mefenacet, mesotrione, metamitron, metazachlor, methabenzthiazuron, metobenzuron, metobromuron, (alpha-) metolachlor, metosulam, metoxuron, metribuzin, metsulfuron (-methyl), molinate, monolinuron, naproanilide, napropamide, neburon, nicosulfuron, norflurazon, orbencarb, oryzalin, oxadiargyl, oxadiazon, oxasulfuron, oxaziclomefone, oxyfluorfen, paraquat, pelargonic acid, pendimethalin, pendralin, pentoxazone, phenmedipham, picolinafen, piperophos, pretilachlor, primisulfuron (-methyl), profluazol, prometryn, propachlor, propanil, propaquizafop, propisochlor, propoxycarbazone (-sodium), propyzamide, prosulfocarb, prosulfuron, pyraflufen (-ethyl), pyrazogyl, pyrazolate, pyrazosulfuron (-ethyl), pyrazoxyfen, pyribenzoxim, pyributicarb, pyridate, pyridatol, pyriftalid, pyriminobac (-methyl), pyrithiobac (-sodium), quinchlorac, quinmerac, quinoclamine, quizalofop (—P-ethyl, —P-tefuryl), rimsulfuron, sethoxydim, simazine, simetryn, sulcotrione, sulfentrazone, sulfometuron (-methyl), sulfosate, sulfosulfuron, tebutam, tebuthiuron, tepraloxydim, terbuthylazine, terbutryn, thenylchlor, thiafluamide, thiazopyr, thidiazimin, thifensulfuron (-methyl), thiobencarb, tiocarbazil, tralkoxydim, tniallate, triasulfuron, tribenuron (-methyl), triclopyr, tridiphane, trifluralin, trifloxysulfuron, triflusulfuron (-methyl), tritosulfuron.","Furthermore suitable for the mixtures are known safeners, for example: AD-67, BAS-145138, benoxacor, cloquintocet (-mexyl), cyometrinil, 2,4-D, DKA-24, dichlormid, dymron, fenclorim, fenchlorazol (-ethyl), flurazole, fluxofenim, furilazole, isoxadifen (-ethyl), MCPA, mecoprop (—P), mefenpyr (-diethyl), MG-191, oxabetrinil, PPG-1292, R-29148.A mixture with other known active compounds, such as fungicides, insecticides, acaricides, nematicides, bird repellents, plant nutrients and agents which improve soil structure, is also possible.","The active compounds can be used as such, in the form of their formulations or in the use forms prepared therefrom by further dilution, such as ready-to-use solutions, suspensions, emulsions, powders, pastes and granules.","They are used in a customary manner, for example by watering, spraying, atomizing or broadcasting.","The active compounds according to the invention can be applied both before and after emergence of the plants.","They can also be incorporated into the soil before sowing.","The amount of active compound used can vary within a relatively wide range.","It depends essentially on the nature of the desired effect.","In general, the amounts used are between 1 g and 10 kg of active compound per hectare of soil surface, preferably between 5 g and 5 kg per ha.","The substances according to the invention have potent microbicidal activity and can be employed for controlling undesirable microorganisms, such as fungi and bacteria, in crop protection and in the protection of materials.","Fungicides can be employed in crop protection for controlling Plasmodiophoromycetes, Oomycetes, Chytridiomycetes, Zygomycetes, Ascomycetes, Basidiomycetes and Deuteromycetes.","Bactericides can be employed in crop protection for controlling Pseudomonadaceae, Rhizobiaceae, Enterobacteriaceae, Corynebacteriaceae and Streptomycetaceae.","Some pathogens causing fungal and bacterial diseases which come under the generic names listed above may be mentioned as examples, but not by way of limitation: Xanthomonas species, such as, for example, Xanthomonas campestris pv.","oryzae; Pseudomonas species, such as, for example, Pseudomonas syringae pv.","lachrymans; Erwinia species, such as, for example, Erwinia amylovora; Pythium species, such as, for example, Pythium ultimum; Phytophthora species, such as, for example, Phytophthora infestans; Pseudoperonospora species, such as, for example, Pseudoperonospora humuli or Pseudoperonospora cubensis; Plasmopara species, such as, for example, Plasmopara viticola; Bremia species, such as, for example, Bremia lactucae; Peronospora species, such as, for example, Peronospora pisi or P. brassicae; Erysiphe species, such as, for example, Erysiphe graminis; Sphaerotheca species, such as, for example, Sphaerotheca fuliginea; Podosphaera species, such as, for example, Podosphaera leucotricha; Venturia species, such as, for example, Venturia inaequalis; Pyrenophora species, such as, for example, Pyrenophora teres or P. graminea (conidia form: Drechslera, syn: Helminthosporium); Cochliobolus species, such as, for example, Cochliobolus sativus (conidia form: Drechslera, syn: Helminthosporium); Uromyces species, such as, for example, Uromyces appendiculatus; Puccinia species, such as, for example, Puccinia recondita; Sclerotinia species, such as, for example, Sclerotinia sclerotiorum; Tilletia species, such as, for example, Tilletia caries; Ustilago species, such as, for example, Ustilago nuda or Ustilago avenae; Pellicularia species, such as, for example, Pellicularia sasakii; Pyricularia species, such as, for example, Pyricularia oryzae; Fusarium species, such as, for example, Fusarium culmorum; Botrytis species, such as, for example, Botrytis cinerea; Septoria species, such as, for example, Septoria nodorum; Leptosphaeria species, such as, for example, Leptosphaeria nodorum; Cercospora species, such as, for example, Cercospora canescens; Alternaria species, such as, for example, Alternaria brassicae; and Pseudocercosporella species, such as, for example, Pseudocercosporella herpotrichoides.","The active compounds according to the invention also have very good fortifying action in plants.","Accordingly, they can be used for mobilizing the defences of the plant against attack by undesirable microorganisms.","In the present context, plant-fortifying (resistance-inducing) substances are to be understood as meaning those substances which are capable of stimulating the defence system of plants such that, when the treated plants are subsequently inoculated with undesirable microorganisms, they show substantial resistance against these microorganisms.","In the present case, undesirable microorganisms are to be understood as meaning phytopathogenic fungi, bacteria and viruses.","Accordingly, the substances according to the invention can be used to protect plants for a certain period after the treatment against attack by the pathogens mentioned.","The period for which protection is provided generally extends over 1 to 10 days, preferably 1 to 7 days, after the treatment of the plants with the active compounds.","The fact that the active compounds are well tolerated by plants at the concentrations required for controlling plant diseases permits the treatment of above-ground parts of plants, of propagation stock and seeds, and of the soil.","The active compounds according to the invention are also suitable for increasing the yield of crops.","In addition, they show reduced toxicity and are well tolerated by plants.","At certain concentrations and application rates, the active compounds according to the invention can also be used as herbicides, for influencing plant growth and for controlling animal pests.","If appropriate, they can also be used as intermediates and precursors for the synthesis of further active compounds.","In the protection of materials, the compounds according to the invention can be employed for protecting industrial materials against infection with, and destruction by, undesired microorganisms.","Industrial materials in the present context are understood as meaning non-living materials which have been prepared for use in industry.","For example, industrial materials which are intended to be protected by active compounds according to the invention from microbial change or destruction can be adhesives, sizes, paper and board, textiles, leather, wood, paints and plastic articles, cooling lubricants and other materials which can be infected with, or destroyed by, microorganisms.","Parts of production plants, for example cooling-water circuits, which may be impaired by the proliferation of microorganisms may also be mentioned within the scope of the materials to be protected.","Industrial materials which may be mentioned within the scope of the present invention are preferably adhesives, sizes, paper and board, leather, wood, paints, cooling lubricants and heat-transfer liquids, particularly preferably wood.","Microorganisms capable of degrading or changing the industrial materials which may be mentioned are, for example, bacteria, fungi, yeasts, algae and slime organisms.","The active compounds according to the invention preferably act against fungi, in particular moulds, wood-discolouring and wood-destroying fungi (Basidiomycetes), and against slime organisms and algae.","Microorganisms of the following genera may be mentioned as examples: Alternaria, such as Alternaria tenuis, Aspergillus, such as Aspergillus niger, Chaetomium, such as Chaetomium globosum, Coniophora, such as Coniophora puetana, Lentinus, such as Lentinus tigrinus, Penicillium, such as Penicillium glaucum, Polyporus, such as Polyporus versicolor, Aureobasidium, such as Aureobasidium pullulans, Sclerophoma, such as Sclerophoma pityophila, Trichoderma, such as Trichoderma viride, Escherichia, such as Escherichia coli, Pseudomonas, such as Pseudomonas aeruginosa, and Staphylococcus, such as Staphylococcus aureus.","Depending on their particular physical and/or chemical properties, the active compounds can be converted into the customary formulations, such as solutions, emulsions, suspensions, powders, foams, pastes, granules, aerosols and microencapsulations in polymeric substances and in coating compositions for seeds, and ULV cool and warm fogging formulations.","These formulations are produced in a known manner, for example by mixing the active compounds with extenders, that is, liquid solvents, liquefied gases under pressure, and/or solid carriers, optionally with the use of surfactants, that is emulsifiers and/or dispersants, and/or foam formers.","If the extender used is water, it is also possible to employ, for example, organic solvents as auxiliary solvents.","Essentially, suitable liquid solvents are: aromatics such as xylene, toluene or alkylnaphthalenes, chlorinated aromatics or chlorinated aliphatic hydrocarbons such as chlorobenzenes, chloroethylenes or methylene chloride, aliphatic hydrocarbons such as cyclohexane or paraffins, for example petroleum fractions, alcohols such as butanol or glycol and their ethers and esters, ketones such as acetone, methyl ethyl ketone, methyl isobutyl ketone or cyclohexanone, strongly polar solvents such as dimethylformamide and dimethyl sulphoxide, or else water.","Liquefied gaseous extenders or carriers are to be understood as meaning liquids which are gaseous at standard temperature and under atmospheric pressure, for example aerosol propellants such as halogenated hydrocarbons, or else butane, propane, nitrogen and carbon dioxide.","Suitable solid carriers are: for example ground natural minerals such as kaolins, clays, talc, chalk, quartz, attapulgite, montmorillonite or diatomaceous earth, and ground synthetic minerals such as finely divided silica, alumina and silicates.","Suitable solid carriers for granules are: for example crushed and fractionated natural rocks such as calcite, marble, pumice, sepiolite and dolomite, or else synthetic granules of inorganic and organic meals, and granules of organic material such as sawdust, coconut shells, maize cobs and tobacco stalks.","Suitable emulsifiers and/or foam formers are: for example nonionic and anionic emulsifiers, such as polyoxyethylene fatty acid esters, polyoxyethylene fatty alcohol ethers, for example alkylaryl polyglycol ethers, alkylsulphonates, alkyl sulphates, arylsulphonates, or else protein hydrolysates.","Suitable dispersants are: for example lignosulphite waste liquors and methylcellulose.","Tackifiers such as carboxymethylcellulose and natural and synthetic polymers in the form of powders, granules or latices, such as gum arabic, polyvinyl alcohol and polyvinyl acetate, or else natural phospholipids such as cephalins and lecithins and synthetic phospholipids can be used in the formulations.","Other possible additives are mineral and vegetable oils.","It is possible to use colorants such as inorganic pigments, for example iron oxide, titanium oxide and Prussian Blue, and organic dyestuffs such as alizarin dyestuffs, azo dyestuffs and metal phthalocyanine dyestuffs, and trace nutrients such as salts of iron, manganese, boron, copper, cobalt, molybdenum and zinc.","The formulations generally comprise between 0.1 and 95% by weight of active compound, preferably between 0.5 and 90%.","The active compounds according to the invention can be used as such or in their formulations, also in a mixture with known fungicides, bactericides, acaricides, nematicides or insecticides, to broaden, for example, the activity spectrum or to prevent development of resistance.","In many cases, synergistic effects are obtained, i.e.","the activity of the mixture is greater than the activity of the individual components.","Examples of suitable mixing components are the following: Fungicides: aldimorph, ampropylfos, ampropylfos-potassium, andoprim, anilazine, azaconazole, azoxystrobin, benalaxyl, benodanil, benomyl, benzamacril, benzamacril-isobutyl, bialaphos, binapacryl, biphenyl, bitertanol, blasticidin-S, bromuconazole, bupirimate, buthiobate, calcium polysulphide, capsimycin, captafol, captan, carbendazim, carboxin, carvon, quinomethionate, chlobenthiazone, chlorfenazole, chloroneb, chloropicrin, chlorothalonil, chlozolinate, clozylacon, cufraneb, cymoxanil, cyproconazole, cyprodinil, cyprofuram, debacarb, dichlorophen, diclobutrazole, diclofluanid, diclomezine, dicloran, diethofencarb, difenoconazole, dimethirimol, dimethomorph, diniconazole, diniconazole-M, dinocap, diphenylamine, dipyrithione, ditalimfos, dithianon, dodemorph, dodine, drazoxolon, edifenphos, epoxiconazole, etaconazole, ethirimol, etridiazole, famoxadon, fenapanil, fenarimol, fenbuconazole, fenfuram, fenitropan, fenpiclonil, fenpropidin, fenpropimorph, fentin acetate, fentin hydroxide, ferbam, ferimzone, fluazinam, flumetover, fluoromide, fluquinconazole, flurprimidol, flusilazole, flusulfamide, flutolanil, flutriafol, folpet, fosetyl-alumninium, fosetyl-sodium, fthalide, fuberidazole, furalaxyl, furametpyr, furcarbonil, furconazole, furconazole-cis, furmecyclox, guazatine, hexachlorobenzene, hexaconazole, hymexazole, imazalil, imibenconazole, iminoctadine, iminoctadine albesilate, iminoctadine triacetate, iodocarb, ipconazole, iprobenfos (IBP), iprodione; irumamycin, isoprothiolane, isovaledione, kasugamycin, kresoxim-methyl, copper preparations, such as: copper hydroxide, copper naphthenate, copper oxychloride, copper sulphate, copper oxide, oxine-copper and Bordeaux mixture, mancopper, mancozeb, maneb, meferimzone, mepanipyrim, mepronil, metalaxyl, metconazole, methasulfocarb, methfuroxam, metiram, metomeclam, metsulfovax, mildiomycin, myclobutanil, myclozolin, nickel dimethyldithiocarbamate, nitrothal-isopropyl, nuarimol, ofurace, oxadixyl, oxamocarb, oxolinic acid, oxycarboxim, oxyfenthiin, paclobutrazole, pefurazoate, penconazole, pencycuron, phosdiphen, picoxystrobin, pimaricin, piperalin, polyoxin, polyoxorim, probenazole, prochloraz, procymidone, propamocarb, propanosine-sodium, propiconazole, propineb, pyraclostrobin, pyrazophos, pyrifenox, pyrimethanil, pyroquilon, pyroxyfur, quinconazole, quintozene (PCNB), sulphur and sulphur preparations, tebuconazole, tecloftalam, tecnazene, tetcyclacis, tetraconazole, thiabendazole, thicyofen, thifluzamide, thiophanate-methyl, thiram, tioxymid, tolclofos-methyl, tolylfluanid, triadimefon, triadimenol, triazbutil, triazoxide, trichlamide, tricyclazole, tridemorph, trifloxystrobin, triflumnizole, triforine, triticonazole, uniconazole, validamycin A, vinclozolin, viniconazole, zarilarnide, zineb, ziram and also Dagger G, OK-8705, OK-8801, α-(1,1-dimethylethyl)-β-(2-phenoxyethyl)-1H-1,2,4-triazole-1-ethanol, α-(2,4-dichlorophenyl)-β-fluoro-β-propyl-1H-1,2,4-triazole-1-ethanol, α-(2,4-dichlorophenyl)-β-methoxy-α-methyl-1H-1,2,4-triazole-1-ethanol, α-(5-methyl-1,3-dioxan-5-yl)-β-[[4-(trifluoromethyl)-phenyl]-methylene]-1H-1,2,4-triazole-1-ethanol, (5RS,6RS)-6-hydroxy-2,2,7,7-tetramethyl-5-(1H-1,2,4-triazol-1-yl)-3-octanone, (E)-α-(methoxyimino)-N-methyl-2-phenoxy-phenylacetamide, isopropyl {2-methyl-1-[[[1-(4-methylphenyl)-ethyl]-amino]-carbonyl]-propyl}-carbamate, 1-(2,4-dichlorophenyl)-2-(1H-1,2,4-triazol-1-yl)-ethanone O-(phenylmethyl)-oxime, 1-(2-methyl-1-naphthalenyl)-1H-pyrrole-2,5-dione, 1-(3,5-dichlorophenyl)-3-(2-propenyl)-2,5-pyrrolidinedione, 1-[(diiodomethyl)-sulphonyl]-4-methyl-benzene, 1-[[2-(2,4-dichlorophenyl)-1,3-dioxolan-2-yl]-methyl]-1H-imidazole, 1-[[2-(4-chlorophenyl)-3-phenyloxiranyl]-methyl]-1H-1,2,4-triazole, 1-[1-[2-[(2,4-dichlorophenyl)-methoxy]-phenyl]-ethenyl]-1H-imidazole, 1-methyl-5-nonyl-2-(phenylmethyl)-3-pyrrolidinole, 2′,6′-dibromo-2-methyl-4′-trifluoromethoxy-4′-trifluoro-methyl-1,3-thiazole-5-carboxanilide, 2,2-dichloro-N-[1-(4-chlorophenyl)-ethyl]-1-ethyl-3-methyl-cyclopropanecarboxamide, 2,6-dichloro-5-(methylthio)-4-pyrimidinyl-thiocyanate, 2,6-dichloro-N-(4-trifluoromethylbenzyl)-benzamide, 2,6-dichloro-N-[[4-(trifluoromethyl)-phenyl]-methyl]-benzamide, 2-(2,3,3-triiodo-2-propenyl)-2H-tetrazole, 2-[(1-methylethyl)-sulphonyl]-5-(trichloromethyl)-1,3,4-thiadiazole, 2-[[6-deoxy-4-O-(4-O-methyl-β-D-glycopyranosyl)-α-D-glucopyranosyl]-amino]-4-methoxy1H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile, 2-aminobutane, 2-bromo-2-(bromomethyl)-pentanedinitrile, 2-chloro-N-(2,3-dihydro-1,1,3-trimethyl-1H-inden-4-yl)-3-pyridinecarboxamide, 2-chloro-N-(2,6-dimethylphenyl)-N-(isothiocyanatomethyl)-acetamide, 2-phenylphenol (OPP), 3,4-dichloro-1-[4-(difluoromethoxy)-phenyl]-1H-pyrrole-2,5-dione, 3,5-dichloro-N-[cyano[(1-methyl-2-propynyl)-oxy]-methyl]-benzamide, 3-(1,1-dimethylpropyl)-1-oxo-1H-indene-2-carbonitrile, 3-[2-(4-chlorophenyl)-5-ethoxy-3-isoxazolidinyl]-pyridine, 4-chloro-2-cyano-N,N-dimethyl-5-(4-methylphenyl)-1H-imidazole-1-sulphonamide, 4-methyl-tetrazolo[1,5-a]quinazolin-5(4H)-one, 8-(1,1-dimethylethyl)-N-ethyl-N-propyl-1,4-dioxaspiro[4.5]decane-2-methaneamine, 8-hydroxyquinoline sulphate, 9H-xanthene-2-[(phenylamino)-carbonyl]-9-carboxylic hydrazide, bis-(1-methylethyl)-3-methyl-4-[(3-methylbenzoyl)-oxy]-2,5-thiophenedicarboxylate, cis-1-(4-chlorophenyl)-2-(1H-1,2,4-triazol-1-yl)-cycloheptanol, cis-4-[3-[4-(1,1-dimethylpropyl)-phenyl-2-methylpropyl]-2,6-dimethyl-morpholine-hydrochloride, ethyl [(4-chlorophenyl)-azo]-cyanoacetate, potassium hydrogen carbonate, methanetetrathiol sodium salt, methyl 1-(2,3-dihydro-2,2-dimethyl-1H-inden-1-yl)-1H-imidazole-5-carboxylate, methyl N-(2,6-dimethylphenyl)-N-(5-isoxazolylcarbonyl)-DL-alaninate, methyl N-(chloroacetyl)-N-(2,6-dimethylphenyl)-DL-alaninate, N-(2,3-dichloro-4-hydroxyphenyl)-1-methyl-cyclohexanecarboxamide, N-(2,6-dimethylphenyl)-2-methoxy-N-(tetrahydro-2-oxo-3-furanyl)-acetamide, N-(2,6-dimethylphenyl)-2-methoxy-N-(tetrahydro-2-oxo-3-thienyl)-acetamide, N-(2-chloro-4-nitrophenyl)-4-methyl-3-nitro-benzenesulphonamide, N-(4-cyclohexylphenyl)-1,4,5,6-tetrahydro-2-pyrimidineamine, N-(4-hexylphenyl)-1,4,5,6-tetrahydro-2-pyrimidineamine, N-(5-chloro-2-methylphenyl)-2-methoxy-N-(2-oxo-3-oxazolidinyl)-acetamide, N-(6-methoxy-3-pyridinyl)-cyclopropanecarboxamide, N-[2,2,2-trichloro-1-[(chloroacetyl)-amino]-ethyl]-benzamide, N-[3-chloro-4,5-bis-(2-propinyloxy)-phenyl]-N′-methoxy-methaneimidamide, N-formyl-N-hydroxy-DL-alanine-sodium salt, O,O-diethyl [2-(dipropylamino)-2-oxoethyl]-ethylphosphoramnidothioate, O-methyl S-phenyl phenylpropylphosphoramidothioate, S-methyl1,2,3-benzothiadiazole-7-carbothioate, spiro[2H]-1-benzopyrane-2,1′(3′H)-isobenzofuran]-3′-one, 4-[(3,4-dimethoxyphenyl)-3-(4-fluorophenyl)-acryloyl]-morpholine.","Bactericides: bronopol, dichlorophen, nitrapyrin, nickel dimethyldithiocarbamate, kasugamycin, octhilinone, furancarboxylic acid, oxytetracyclin, probenazole, streptomycin, tecloftalam, copper sulphate and other copper preparations.","Insecticides/acaricides/nematicides: abamectin, acephate, acetamiprid, acrinathrin, alanycarb, aldicarb, aldoxycarb, alpha-cypermethrin, alphamethrin, amitraz, avermectin, AZ 60541, azadirachtin, azamethiphos, azinphos A, azinphos M, azocyclotin, Bacillus popilliae, Bacillus sphaericus, Bacillus subtilis, Bacillus thuringiensis, baculoviruses, Beauveria bassiana, Beauveria tenella, bendiocarb, benfuracarb, bensultap, benzoximate, betacyfluthrin, bifenazate, bifenthrin, bioethanomethrin, biopermethrin, bistrifluron, BPMC, bromophos A, bufencarb, buprofezin, butathiofos, butocarboxim, butylpyridaben, cadusafos, carbaryl, carbofuran, carbophenothion, carbosulfan, cartap, chloethocarb, chlorethoxyfos, chlorfenapyr, chlorfenvinphos, chlorfluazuron, chlormephos, chlorpyrifos, chlorpyrifos M, chlovaporthrin, chromafenozide, cis-resmethrin, cispermnethrin, clocythrin, cloethocarb, clofentezine, clothianidine, cyanophos, cycloprene, cycloprothrin, cyfluthrin, cyhalothrin, cyhexatin, cypermethrin, cyromazine, deltamethrin, demeton M, demeton S, demeton-S-methyl, diafenthiuron, diazinon, dichlorvos, dicofol, diflubenzuron, dimethoat, dimethylvinphos, diofenolan, disulfoton, docusat-sodium, dofenapyn, eflusilanate, emamectin, empenthrin, endosulfan, Entomopfthora spp., esfenvalerate, ethiofencarb, ethion, ethoprophos, etofenprox, etoxazole, etrimfos; fenamiphos, fenazaquin, fenbutatin oxide, fenitrothion, fenothiocarb, fenoxacrim, fenoxycarb, fenpropathrin, fenpyrad, fenpyrithrin, fenpyroximate, fenvalerate, fipronil, fluazinam, fluazuron, flubrocythrinate, flucycloxuron, flucythrinate, flufenoxuron, flumethrin, flutenzine, fluvalinate, fonophos, fosmethilan, fosthiazate, fubfenprox, furathiocarb, granulosis viruses, halofenozide, HCH, heptenophos, hexaflumuron, hexythiazox, hydroprene, imidacloprid, indoxacarb, isazofos, isofenphos, isoxathion, ivermectin, nuclear polyhedrosis viruses, lambda-cyhalothrin, lufenuron, malathion, mecarbam, metaldehyde, methamidophos, Metharhizium anisopliae, Metharhizium flavoviride, methidathion, methiocarb, methoprene, methomyl, methoxyfenozide, metolcarb, metoxadiazone, mevinphos, milbemectin, milbemycin, monocrotophos, naled, nitenpyram, nithiazine, novaluron, omethoate, oxamyl, oxydemethon M, Paecilomyces fumosoroseus, parathion A, parathion M, permethrin, phenthoate, phorat, phosalone, phosmet, phosphamidon, phoxim, pirimicarb, pirimiphos A, pirimiphos M, profenofos, promecarb, propargite, propoxur, prothiofos, prothoat, pymetrozine, pyraclofos, pyresmethrin, pyrethrum, pyridaben, pyridathion, pyrimidifen, pyriproxyfen, quinalphos, ribavirin, salithion, sebufos, silafluofen, spinosad, spirodiclofen, sulfotep, sulprofos, tau-fluvalinate, tebufenozide, tebufenpyrad, tebupirimiphos, teflubenzuron, tefluthrin, temephos, temivinphos, terbufos, tetrachlonrinphos, tetradifon theta-cypermethrin, thiacloprid, thiamethoxam, thiapronil, thiatriphos, thiocyclam hydrogen oxalate, thiodicarb, thiofanox, thuringiensin, tralocythrin, tralomethrin, triarathene, triazamate, triazophos, triazuron, trichlophenidine, trichlorfon, triflumuron, trimethacarb, vamidothion, vaniliprole, Verticillium lecanii, YI5302 zeta-cypermethfin, zolaprofos (1R-cis)-[5-(phenylmethyl)-3-furanyl]-methyl-3-[(dihydro-2-oxo-3(2H)-furanylidene)-methyl]-2,2-dimethylcyclopropanecarboxylate, (3-phenoxyphenyl)-methyl-2,2,3,3-tetramethylcyclopropanecarboxylate, 1-[(2-chloro-5-thiazolyl)methyl]tetrahydro-3,5-dimethyl-N-nitro-1,3,5-triazine-2(1H)-imine, 2-(2-chloro-6-fluorophenyl)-4-[4-(1,1-dimethylethyl)phenyl]-4,5-dihydro-oxazole, 2-(acetyloxy)-3-dodecyl-1,4-naphthalenedione, 2-chloro-N-[[[4-(1-phenylethoxy)-phenyl]-amino]-carbonyl]-benzamide, 2-chloro-N-[[[4-(2,2-dichloro-1,1-difluoroethoxy)-phenyl]-amino]-carbonyl]-benzamide, 3-methylphenyl propylcarbamate, 4-[4-(4-ethoxyphenyl)-4-methylpentyl]-1-fluoro-2-phenoxy-benzene, 4-chloro-2-(1,1-dimethylethyl)-5-[[2-(2,6-dimethyl-4-phenoxyphenoxy)ethyl]thio]-3(2H)-pyridazinone, 4-chloro-2-(2-chloro-2-methylpropyl)-5-[(6-iodo-3-pyridinyl)methoxy]-3(2H)-pyridazinone, 4-chloro-5-[(6-chloro-3-pyridinyl)methoxy]-2-(3,4-dichlorophenyl)-3(2H)-pyridazinone, Bacillus thuringiensis strain EG-2348, [2-benzoyl-1-(1,1-dimethylethyl)-hydrazinobenzoic acid, 2,2-dimethyl-3-(2,4-dichlorophenyl)-2-oxo-1-oxaspiro[4.5]dec-3-en4-yl butanoate, [3-[(6-chloro-3-pyridinyl)methyl]-2-thiazolidinylidene]-cyanamide, dihydro-2-(nitromethylene)-2H-1,3-thiazine-3(4H)-carboxaldehyde, ethyl [2-[[1,6-dihydro-6-oxo-1-(phenylmethyl)-4-pyridazinyl]oxy]ethyl]-carbamate, N-(3,4,4-trifluoro-1-oxo-3-butenyl)-glycine, N-(4-chlorophenyl)-3-[4-(difluoromethoxy)phenyl]-4,5-dihydro4-phenyl-1H-pyrazole-1-carboxamide, N-[(2-chloro-5-thiazolyl)methyl]-N′-methyl-N″-nitro-guanidine, N-methyl-N′-(1-methyl-2-propenyl)-1,2-hydrazinedicarbothioamide, N-methyl-N′-2-propenyl-1,2-hydrazinedicarbothioamide, O,O-diethyl [2-(dipropylamino)-2-oxoethyl]-ethylphosphoramidothioate, N-cyanomethyl-4-trifluoromethyl-nicotinamide, 3,5-dichloro-1-(3,3-dichloro-2-propenyloxy)-4-[3-(5-trifluoromethylpyridine-2-yloxy)-propoxy]-benzene.","A mixture with other known active compounds, such as herbicides, or with fertilizers and growth regulators, is also possible.","In addition, the compounds of the formula (I) according to the invention also have very good antimycotic activity.","They have a very broad antimycotic activity spectrum in particular against dermatophytes and yeasts, moulds and diphasic fungi (for example against Candida species, such as Candida albicans, Candida glabrata), and Epidermophyton floccosum, Aspergillus species, such as Aspergillus niger and Aspergillus fumigatus, Trichophyton species, such as Trichophyton mentagrophytes, Microsporon species such as Microsporon canis and audouinii.","The list of these fungi by no means limits the mycotic spectrum covered, but is only for illustration.","The active compounds can be used as such, in the form of their formulations or the use forms prepared therefrom, such as ready-to-use solutions, suspensions, wettable powders, pastes, soluble powders, dusts and granules.","Application is carried out in a customary manner, for example by watering, spraying, atomizing, broadcasting, dusting, foaming, spreading, etc.","It is furthermore possible to apply the active compounds by the ultra-low-volume method, or to inject the active compound preparation or the active compound itself into the soil.","It is also possible to treat the seeds of the plants.","When using the active compounds according to the invention as fungicides, the application rates can be varied within a relatively wide range, depending on the kind of application.","For the treatment of parts of plants, the active compound application rates are generally between 0.1 and 10,000 g/ha, preferably between 10 and 1000 g/ha.","For seed dressing, the active compound application rates are generally between 0.001 and 50 g per kilogram of seed, preferably between 0.01 and 10 g per kilogram of seed.","For the treatment of the soil, the active compound application rates are generally between 0.1 and 10,000 g/ha, preferably between 1 and 5000 g/ha.","The preparation and the use of the active compounds according to the invention is illustrated by the examples below.","PREPARATION EXAMPLES Example I-1-a-1 1.65 g (14.4 mmol) of potassium tert-butoxide are initially charged in 30 ml of anhydrous dimethylformamide (DMF), and 3.5 g (9.6 mmol) of the compound according to Example (II-1) in 10 ml of anhydrous DMF are added dropwise.","The mixture is stirred at 50° C. for 3 h. 100 ml of ice-water are then poured into the reaction solution.","This mixture is poured into 600 ml of cold 1N HCl solution.","The precipitate is filtered off with suction and dried.","Yield: 2.9 g (90% of theory), m.p.","139-141° C. The following compounds of the formula (I-1-a) were prepared analogously to Example (I-1-a-1) and in accordance with the general statements about the preparation: (I-1-a) Ex.","No.","X Y B A Q1 Q2 m.p.° C. I-1-a-2 CH3 4-Cl—C6H4 —(CH2)5— H H 105-107° C. I-1-a-3 C2H5 4-Cl—C6H4 —(CH2)5— H H 121-123° C. I-1-a-4 CH3 4-Cl—C6H4 —(CH2)2—O—(CH2)4— H H 130 I-1-a-5 CH3 4-CF3—C6H4 —(CH2)5— H H Oil I-1-a-6 CH3 4-Cl—C6H4 H —(CH2)4— H 145-148 I-1-a-7 CH3 4-CH3—C6H4 —(CH2)5— H H 85 I-1-a-8 CH3 4-Cl—C6H4—O—C6H4 —(CH2)5— H H 75-77 I-1-a-9 CH3 4-Cl—C6H4 —(CH2)2—CHCH3—(CH2)2— H H 142-144 I-1-a-10 CH3 4-Cl—C6H4 —(CH2)2—CHOCH3—(CH2)2— H H 127 I-1-a-11 CH3 4-Cl—C6H4 H C6H5— H H 130 I-1-a-12 C2H5 4-Cl—C6H4 —(CH2)2—O—(CH2)2— H H 210 I-1-a-13 CH3 4-Cl—C6H4 H C6H11— H H 105 I-1-a-14 CH3 4-Cl—C6H4 —(CH2)2—CH—C3H7—(CH2)2— H H oil I-1-a-15 CH3 4-Cl—C6H4 —(CH2)2—CH-t-C4H9—(CH2)2— H H oil I-1-a-16 CH3 4-Cl—C6H4 —(CH2)2—CH—C2H5—(CH2)2— H H oil I-1-a-17 H 4-Cl—C6H4 —(CH2)5— H H 185 Example I-1-b-1 500 mg (1.33 mmol) of the compound according to Example I-1-a-2 are initially charged in 5 ml of absolute dichloromethane, and 0.27 ml (2.0 mmol) of triethylamine is added.","At 0° C., 0.30 g (1.74 mmol) of 6-chloronicotinyl chloride is added, and the mixture is stirred at room temperature for 2 h. A further 0.15 g of chloronicotinyl chloride and 0.13 ml of triethylamine are then added.","The reaction mixture is washed with 10% citric acid solution, the aqueous phase is extracted with dichloromethane, and the organic phase is then washed with 1N aqueous sodium hydroxide solution and the aqueous phase is extracted with dichloromethane.","The organic phase is dried and the solvent is distilled off.","The residue is stirred with petroleum ether and filtered and then dried.","Yield: 0.7 g (100% of theory), m.p.","133-134° C. The following compounds of the formula (I-1-b) were obtained analogously to Example (1-1-b-1) and in accordance with the general statements about the preparation: (I-1-b) Ex.","No.","X Y B A Q1 Q2 R1 m.p.° C. I-1-b-2 CH3 4-Cl—C6H4 —(CH2)5— H H H5C2—O—CH2— 135 I-1-b-3 CH3 4-Cl—C6H4 —(CH2)5— H H i-C3H7 oil I-1-b-4 CH3 4-Cl—C6H4 —(CH2)5— H H 4-Cl—C6H4 129-130 I-1-b-5 CH3 4-Cl—C6H4 —(CH2)5— H H Oil Example I-1-c-1 500 mg (1.5 mmol) of the compound according to Example (I-1-a-1) are initially charged in 5 ml of absolute dichloromethane, and 0.3 ml (2.25 mmol) of triethylamine is added.","At 0° C., 0.225 g (1.95 mmol) of ethyl chloroformate is added, and the mixture is stirred at room temperature for 2 h. The reaction solution is washed with 10% strength citric acid and the aqueous phase is extracted with dichloromethane (CH2Cl2).","The organic phase is washed with 1N sodium hydroxide solution, dried and concentrated.","Yield: 0.45 g (75% of theory), 132-134° C. The following compounds of the formula (I-1-c) were obtained analogously to Example (I-1-c-1) and in accordance with the general statements about the preparation: (I-1-c) Ex.No.","X Y A B Q1 Q2 L M R2 m.p.° C. I-1-c-2 CH3 4-Cl—C6H4 —(CH2)5— H H O O C25 oil I-1-c-3 C2H5 4-Cl—C6H4 —(CH2)5— H H O O C25 oil I-1-c-4 CH3 4-Cl—C6H4 —(CH2)5— H H O O C65 139-140 I-1-c-5 CH3 4-Cl—C6H4 —(CH2)5— H H O S C65—CH2 144 I-1-c-6 CH3 4-Cl—C6H4 —(CH2)5— H H O O C65—CH2 137 Example I-1-d-1 0.5 g (1.33 mmol) of the compound according to Example I-1-a-2 is initially charged in 5 ml of anhydrous dichloromethane, and 0.27 ml (2.0 mmol) of triethylamine is added.","With ice-cooling, 0.135 ml (1.74 mmol) of methanesulphonyl chloride is added.","The mixture is stirred at room temperature for 2 h. The reaction mixture is extracted with 10% strength citric acid, the aqueous phase is washed with dichloromethane, the organic phase is extracted with 1N sodium hydroxide solution and the aqueous phase is washed with dichloromethane, dried and concentrated.","Yield: 0.6 g (100% of theory), m.p.","134° C. 1H-NMR (d6-DMSO, 400 MHz): δ=1.02-1.75 (m, 10H, cyc-hexyl-CH2), 2.30 (s, 3H, thiazolyl-CH3), 3.05 (s, 2H, CH2—CO), 3.61 (s, 3H, SO2—CH3), 7.54 (d, 2H, Ar-H), 7.86 (d, 2H, Ar-H) ppm.","Example I-1-g-1 0.5 g (1.33 mmol) of the compound according to Example I-1-a-2 and 0.27 ml (2.0 mmol) of triethylamine are dissolved in 5 ml of anhydrous dichloromethane, and 0.26 g (1.73 mmol) of morpholine N-carbonyl chloride are added with ice-cooling.","The mixture is stirred at room temperature overnight.","The reaction mixture is extracted with 10% strength citric acid, the aqueous phase is washed with dichloromethane, the organic phase is extracted with 1N sodium hydroxide solution and the aqueous phase is washed with dichloromethane, dried and concentrated.","Yield: 0.69 g (93% of theory), wax 1H-NMR (d6-DMSO, 400 MHz): δ=1.02-1.74 (m, 10H, cyc-hexyl-CH2), 2.30 (s, 3H, thiazolyl-CH3), 3.01 (s, 2H, CH2—CO), 3.05-3.79 (m, 8H, morpholine-CH2), 7.54 (d, 2H, Ar-H), 7.85 (d, 2H, Ar-H) ppm.","Example II-1 17.3 g (30 mmol) of the compound according to Example XIV-1 are initially charged in 200 ml of anhydrous acetone, and 6.8 g (49 mmol) of potassium carbonate and 20.5 g (147 mmol) of methyl iodide are added.","The mixture is stirred under reflux for 16 h. The solution is filtered, the solvent is distilled off and the residue is purified by silica gel chromatography (dichloromethane:petroleum ether, 4:1→dichloromethane→dichloromethane:ethyl acetate, 30:1→10:1).","Yield: 3.5 g (31% of theory), oil 1H-NMR (d6-DMSO, 400 MHz): δ=1.11 (s, 6H, 2-CH3-C-aliph.","), 2.31 (s, 3H, CH3-C-heteroarom.)","3.50 (s, 3H, CH3—O), 7.50 (d, 2H, 2-CH-arom.","), 7.84 (d, 2H, 2-CH-arom) ppm.","Example XIV-1 A solution of 12.7 g (45 mmol) of methyl 4-[2-(4-chlorophenyl)-5-methyl]-thiazolyl-acetate in 15 ml of tetrahydrofuran is, at −15° C., added dropwise to a solution of 25 ml of LDA* solution (2 molar) in 50 ml of anhydrous tetrahydrofuran, and the mixture is stirred at 0° C. for 60 min.","At −15° C., a solution of 3.8 g (30 mmol) of 2,2-dimethylsuccinic anhydride in 10 ml of anhydrous tetrahydrofuran is then added dropwise.","The solution is stirred at room temperature for two hours, 75 ml of water and 20 g of ammonium chloride are then added and the solution is acidified using concentrated hydrochloric acid.","The intermediate is extracted with ether and the solvent is distilled off.","The residue is boiled under reflux with 25 g KOH and 170 ml of water for one day.","The mixture is cooled, acidified with conc.","hydrochloric acid and extracted with ether.","The crude product is directly, without further characterization and purification, converted into the compound II-1.Yield: 17.5 g (100% of theory), oil *LDA =lithium diisopropylamide The following compounds of the formula (II) were obtained analogously to Example (II-1) and in accordance with the general statements about the preparation: (II) Ex.","No.","X Y B A Q1 Q2 R8 m.p.° C. II-2 CH3 4-Cl—C6H4 —(CH2)5— H H CH3 oil II-3 C2H5 4-Cl—C6H4 —(CH2)5— H H CH3 oil II-4 CH3 4-Cl—C6H4 —(CH2)2—O—(CH2)2— H H CH3 oil II-5 CH3 4-CF3—C6H4 —(CH2)5— H H CH3 oil II-6 CH3 4-Cl—C6H4 H —(CH2)4— H CH3 oil II-7 CH3 4-CH3—C6H4 —(CH2)5— H H CH3 oil II-8 CH3 4-Cl—C6H4—O—C6H4 —(CH2)5— H H CH3 oil II-9 CH3 4-Cl—C6H4 —(CH2)2—CHCH3—(CH2)2— H H CH3 oil II-10 CH3 4-Cl—C6H4 —(CH2)2—CHOCH3—(CH2)2— H H CH3 oil II-11 CH3 4-Cl—C6H4 H C6H5— H H CH3 oil II-12 C2H5 4-Cl—C6H4 —(CH2)2—O—(CH2)2— H H CH3 oil II-13 CH3 4-Cl—C6H4 H C6H11 H H CH3 oil II-14 CH3 4-Cl—C6H4 —(CH2)2—CH—C3H7—(CH2)2— H H CH3 oil II-15 CH3 4-Cl—C6H4 —(CH2)2—CH-t-C4H9—(CH2)2— H H CH3 oil II-16 CH3 4-Cl—C6H4 —(CH2)2—CH—C2H5—(CH2)2— H H CH3 oil II-17 CH3 4-Cl—C6H4 —(CH2)5— H H CH3 oil The following compounds of the formula (XIV) were obtained analogously to Example (XIV-1) and in accordance with the general statements about the preparation: (XIV) Ex.","No.","X Y B A Q1 Q2 m.p.° C. XIV-2 CH3 4-Cl—C6H4 —(CH2)5— H H wax XIV-3 C2H5 4-Cl—C6H4 —(CH2)5— H H wax XIV-4 CH3 4-Cl—C6H4 —(CH2)2—O—(CH2)2— H H oil XIV-5 CH3 4-CF3—C6H4 —(CH2)5— H H oil XIV-6 CH3 4-Cl—C6H4 H —(CH2)4— H oil XIV-7 CH3 4-CH3—C6H4 —(CH2)5— H H oil XIV-8 CH3 4-Cl—C6H4—O—C6H4 —(CH2)5— H H oil XIV-9 CH3 4-Cl—C6H4 —(CH2)2—CHCH3—(CH2)2— H H oil XIV-10 CH3 4-Cl—C6H4 —(CH2)2—CHOCH3—(CH2)2— H H oil XIV-11 CH3 4-Cl—C6H4 H C6H5— H H oil XIV-12 C2H5 4-Cl—C6H4 —(CH2)2—O—(CH2)2— H H oil XIV-13 CH3 4-Cl—C6H4 H C6H11— H H oil XIV-14 CH3 4-Cl—C6H4 —(CH2)2—CH—C3H7—(CH2)2— H H oil XIV-15 CH3 4-Cl—C6H4 —(CH2)2—CH-t-C4H9—(CH2)2— H H oil XIV-16 CH3 4-Cl—C6H4 —(CH2)2—CH—C2H5—(CH2)2— H H oil XIV-17 H 4-Cl—C6H4 —(CH2)5— H H oil Example 1-2-a-1 2.16 g (18.1 mmol) of potassium tert-butoxide are initially charged in 20 ml of anhydrous dimethylformamide, and 4.6 g (12.1 mmol) of the compound according to Example III-1 in 2 ml of anhydrous dimethylformamide are added dropwise.","The mixture is stirred at 50° C. for 2 hours.","The reaction solution is poured into 100 ml of ice-water, and this mixture is stirred into 500 ml of cold 1N hydrochloric acid solution.","The precipitate is filtered off with suction and taken up in dichloromethane.","The solvent is distilled off, and the residue is then purified by silica gel chromatography (cyclohexane/ethyl acetate, 5:1).","Yield: 1.4 g (13% of theory), m.p.","165-167° C. The following compounds of the formula (I-2-a) were obtained analogously to Example (I-2-a-1) and in accordance with the general statements about the preparation: (I-2-a) Ex.","No.","X Y A B Q1 Q2 Q3 Q4 m.p.° C. I-2-a-2 CH3 4-Cl—C6H4 H H H H H H 106 I-2-a-3 CH3 4-Cl—C6H4 CH3 CH3 H H H H oil I-2-a-4 CH3 4-Cl—C6H4 H H —(CH2)4— H H 122 I-2-a-5 CH3 4-Cl—C6H4 H H 4-Cl—C6H4 H H H 195-197 Example 1-2-b-1 500 mg (1.44 mmol) of the compound according to Example I-2-a-1 are initially charged in 5 ml of anhydrous dichloromethane, and 0.29 ml (2.16 mmol) of triethylamine is added.","At 0° C., 0.33 g (1.87 mmol) of 6-chloronicotinyl chloride is added, and the mixture is stirred at room temperature for 2 h. The reaction mixture is extracted with 10% citric acid, the aqueous phase is washed with dichloromethane, the organic phase is extracted with 1N sodium hydroxide solution and the aqueous phase is washed with dichloromethane, dried and concentrated.","Yield: 0.7 g (100% of theory), m.p.","127-130° C. The following compounds of the formula (I-2-b) were obtained analogously to Example (I-2-b-1) and in accordance with the general statements about the preparation: (I-2-b) Ex.","No.","X Y A B Q1 Q2 Q3 Q4 R1 m.p.° C. I-2-b-2 CH3 4-Cl—C6H4 H H CH3 CH3 H H i-C3H7— 96 I-2-b-3 CH3 4-Cl—C6H4 H H CH3 CH3 H H H5C2—O—CH2— 142 I-2-b-4 CH3 4-Cl—C6H4 H H CH3 CH3 H H 4-Cl—C6H4 123-125 I-2-b-5 CH3 4-Cl—C6H4 H H CH3 CH3 H H 125 Example 1-2-c-1 600 mg (1.7 mmol) of the compound according to Example I-2-a-1 are initially charged in 5 ml of anhydrous dichloromethane, and 0.35 ml (2.60 mmol) of triethylamine is added.","At 0° C., 0.255 g (2.21 mmol) of ethyl chloroformate is added, and the mixture is stirred at room temperature for 2 hours.","The solution is washed with 10% strength citric acid and the aqueous phase is extracted with dichloromethane.","The organic phase is washed with 1N sodium hydroxide solution, dried and concentrated.","Yield: 0.62 g (87% of theory), m.p.","82-86° C. The following compounds of the formula (I-2-c) were obtained analogously to Example (I-2-c-1) and in accordance with the general statements about the preparation: (I-2-c) Ex.","No.","X Y A B Q1 Q2 Q3 Q4 M R2 m.p.° C. I-2-c-2 CH3 4-Cl—C6H4 H H CH3 CH3 H H O C6H5— 123-125 I-2-c-3 CH3 4-Cl—C6H4 H H CH3 CH3 H H O C6H5—CH2 129-131 I-2-c-4 CH3 4-Cl—C6H4 H H CH3 CH3 H H S C6H5—CH2 94 Example No.","I-2-d-1 500 mg (1.44 mmol) of the compound according to Example I-2-a-1 are initially charged in 5 ml of anhydrous dichloromethane, and 0.29 ml (2.16 mmol) of triethylamine is added.","At 0° C., 0.22 g (1.87 mmol) of methylsulphonyl chloride is added, and the mixture is stirred at room temperature for 2 h. The reaction mixture is extracted with 10% strength citric acid, the aqueous phase is washed with dichloromethane, the organic phase is extracted with 1N NaOH and the aqueous phase is washed with dichloromethane, dried and concentrated.","Yield: 0.65 g (100% of theory), wax 1H-NMR (d6-DMSO, 400 MHz): δ=1.17 (s, 6H, cyc-hexyl-CH3), 2.25 (s, 3H, thiazolyl-CH3), 2.48 (s, 2H, cyc-hexyl-CH2), 2.86 (s, 2H, cyc-hexyl-CH2), 3.21 (s, 3H, SO2—CH3), 7.52 (d, 2H, Ar-H) 7.83 (d, 2H, Ar-H) ppm.","Example No.","I-2-g-1 500 mg (1.44 mmol) of the compound according to Example I-2-a-1 are initially charged in 5 ml of anhydrous dichloromethane, and 0.29 ml (2.16 mmol) of triethylamine is added.","At 0° C., 0.27 g (1.87 mmol) of morpholinecarbonyl chloride is added, and the mixture is stirred at room temperature for 2 h. The reaction mixture is extracted with 10% strength citric acid, the aqueous phase is washed with dichloromethane, the organic phase is extracted with 1N sodium hydroxide solution and the aqueous phase is washed with dichloromethane, dried and concentrated.","Yield: 0.65 g (100% of theory), m.p.","128° C. 1H-NMR (d6-DMSO, 400 MHz): δ=1.13 (s, 6H, cyc-hexyl-CH3), 2.21 (s, 3H, thiazolyl-CH3), 2.46 (s, 2H, cyc-hexyl-CH2), 2.71 (s, 2H, cyc-hexyl-CH2), 3.06-3.80 (m, 8H, morpholine-CH2), 7.53 (d, 2H, Ar-H), 7.84 (d, 2H, Ar-H) ppm.","Example III-1 17.0 g (30 mmol) of crude product of the compound according to Example XIX-1 are initially charged in 200 ml of anhydrous acetone, and 6.42 g (46.5 mmol) of potassium carbonate and 19.5 g (139.5 mmol) of methyl iodide are added.","The mixture is stirred under reflux for 16 h. The mixture is filtered and concentrated.","The residue is purified by silica gel chromatography (dichloromethane:petroleum ether, 4:1→dichloromethane→dichloromethane:ethyl acetate, 30:1→10:1) Yield: 4.6 g (16% of theory), oil 1H-NMR (400 MHz, d6-DMSO): δ=1.03 (s, 6H, 2—CH3-C-aliph.","), 2.32 (s, 3H, CH3—C-arom.)","7.50 (d, 2H, Ar-H.), 7.81 (d, 2H, Ar-H) ppm.","Example XIX-1 A solution of 12.7 g (45 mmol) of methyl 4-[2-(4-chlorophenyl)-5-methyl]-thiazolyl-acetate in 15 ml of anhydrous tetrahydrofuran is, at −15° C., added dropwise to a solution of 25 ml of LDA solution (2 molar) in 50 ml of anhydrous tetrahydrofuran, and the mixture is stirred at 0° C. for 60 min.","At −15° C., a solution of 4.26 g (30 mmol) of 3,3-dimethylglutaric anhydride in 10 ml of tetrahydrofuran is then added dropwise.","The solution is stirred at room temperature for 2 hours, 75 ml of water and 20 g of ammonium chloride are then added and the mixture is then acidified using concentrated hydrochloric acid.","The intermediate is extracted with ether and the solvent is distilled off.","The residue is boiled under reflux with 25 g of potassium hydroxide and 170 ml of water for one day.","The mixture is cooled, acidified with concentrated hydrochloric acid and extracted with ether.","The crude product is directly, without purification, reacted further.","Yield: 17 g (100% of theory), oil The following compounds of the formula (III) were obtained analogously to Example (III-1) and in accordance with the general statements about the preparation: (III) Ex.","No.","X Y A B Q1 Q2 Q3 Q4 R8 m.p.° C. III-2 CH3 4-Cl—C6H4 H H H H H H CH3 oil III-3 CH3 4-Cl—C6H4 CH3 CH3 H H H H CH3 oil III-4 CH3 4-Cl—C6H4 H H —(CH2)4— H H CH3 oil III-5 CH3 4-Cl—C6H4 H H 4-Cl—C6H4 H H H CH3 oil The following compounds of the formula (XIX) were obtained analogously to Example (XIX-1) and in accordance with the general statements about the preparation: (XIX) Ex.","No.","X Y A B Q1 Q2 Q3 Q4 m.p.° C. XIX-2 CH3 4-Cl—C6H4 H H H H H H oil XIX-3 CH3 4-Cl—C6H4 CH3 CH3 H H H H oil XIX-4 CH3 4-Cl—C6H4 H H —(CH2)4— H H oil XIX-5 CH3 4-Cl—C6H4 H H 4-Cl—C6H4 H H H oil Use Examples Example A Meloidogyne Test Solvent: 30 parts by weight of dimethylformamide Emulsifier: 1 part by weight of alkylaryl polyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amounts of solvent and emulsifier, and the concentrate is diluted with water to the desired concentration.","Containers are filled with sand, solution of active compound, Meloidogyne incognita egg/larvae suspension and lettuce seeds.","The lettuce seeds germinate the plants develop.","On the roots, galls are formed.","After the desired period of time, the nematicidal action is determined in % by the gall formation.","100% means that no galls have been found; 0% means that the number of galls on the treated plants corresponds to that of the untreated control.","In this test, for example, the following compounds of the preparation examples showed good activity: Concentration of active Active compounds compound in ppm Kill rate in % after 14 d Ex.","I-1-c-3 20 100 Ex.","I-1-a-1 20 95 Example B Myzus Test (Systemic Action) Solvent: 7 parts by weight of dimethylformamide Emulsifier: 2 parts by weight of alkylaryl polyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amounts of solvent and emulsifier, and the concentrate is diluted with emulsifier-containing water to the desired concentration.","Savoy cabbage (Brassica oleracea) which is heavily infested by the green peach aphid (Myzus persicae) is watered with in each case 10 ml of preparation of active compound of the desired concentration such that the preparation of active compound penetrates into the soil without wetting the seedling.","The active compound is taken up by the roots and transferred into the seedling.","After the desired period of time, the kill in % is determined.","100% means that all aphids have been killed; 0% means that none of the aphids have been killed.","In this test, for example, the following compounds of the preparation examples show good activity: Concentration of active Active compounds compound in ppm Kill rate in % after 10 d Ex.","I-1-c-2 20 95 Example C Phaedon Larvae Test Solvent: 7 parts by weight of dimethylformamide Emulsifier: 2 parts by weight of alkylaryl polyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amounts of solvent and emulsifier, and the concentrate is diluted with emulsifier-containing water to the desired concentration.","Cabbage leaves (Brassica oleracea) are treated by being dipped into the preparation of active compound of the desired concentration and are populated with larvae of the mustard beetle (Phaedon cochleariae) while the leaves are still moist.","After the desired period of time, the kill in % is determined.","100% means that all beetle larvae have been killed; 0% means that none of the beetle larvae have been killed.","In this test, for example, the following compounds of the preparation examples show good activity: Concentration of active Active compounds compound in ppm Kill rate in % after 7 d Ex.","I-1-c-1 500 100 Ex.","I-2-c-1 500 100 Example D Spodoptera Frugiperda Test Solvent: 7 parts by weight of dimethylfornamide Emulsifier: 2 parts by weight of alkylaryl polyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amounts of solvent and emulsifier, and the concentrate is diluted with emulsifier-containing water to the desired concentration.","Cabbage leaves (Brassica oleracea) are treated by being dipped into the preparation of active compound of the desired concentration and are populated with caterpillars of the armyworm (Spodoptera frugiperda) while the leaves are still moist.","After the desired period of time, the kill in % is determined.","100% means that all caterpillar larvae have been killed; 0% means that none of the caterpillar have been killed.","In this test, for example, the following compounds of the preparation examples show good activity: Concentration of active Active compound compound in ppm Kill rate in % after 7 d Ex.","I-1-a-2 500 100 Ex.","I-2-a-1 500 100 Ex.","I-1-c-1 500 100 Ex.","I-2-c-1 500 100 Example E Tetranychus test (OP-Resistant/Dip Treatment) Solvent: 30 parts by weight of dimethylformamide Emulsifier: 1 part by weight of alkylaryl polyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amounts of solvent and emulsifier, and the concentrate is diluted with emulsifier-containing water to the desired concentration.","Bean plants (Phaseolus vulgaris) which are heavily infested by all stages of the greenhouse red spider mite (Tetranychus urticae) are dipped into a preparation of active compound of the desired concentration.","After the desired period of time, the effect in % is determined.","100% means that all spider mites have been killed; 0% means that none of the spider mites have been killed.","In this test, for example, the following compounds of the preparation examples show good activity: Concentration of active Active compound compound in ppm Kill rate in % after 7 d Ex.","I-1-c-3 100 98 Ex.","I-1-a-1 100 100 Ex.","I-2-a-2 100 99 Ex.","I-2-a-1 100 98 Ex.","I-1-c-1 100 95 Ex.","I-2-c-1 100 100 Example F1 Post-Emergence Test Solvent: 5 parts by weight of acetone Emulsifier: 1 part by weight of alkylaryl polyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amount of solvent, the stated amount of emulsifier is added and the concentrate is diluted with water to the desired concentration.","Test plants of a height of 5-15 cm are sprayed with the preparation of active compound such that the particular amounts of active compound desired are applied per unit area.","The concentration of the spray liquor is chosen such that the particular amounts of active compound desired are applied in 1000 l of water/ha.","After three weeks, the degree of damage to the plants is rated in % damage in comparison to the development of the untreated control.","The figures denote: 0% =no effect (like untreated control) 100% =total destruction Example F2 Pre-Emergence Test Solvent: 5 parts by weight of acetone Emulsifier: 1 part by weight of alkylaryl polyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amount of solvent, the stated amount of emulsifier is added and the concentrate is diluted with water to the desired concentration.","Seeds of the test plants are sown in normal soil.","After about 24 hours, the soil is sprayed with the preparation of active compound such that the particular amounts of active compound desired are applied per unit area.","The concentration of the spray liquor is chosen such that the particular amounts of active compound desired are applied in 1000 l of water/ha.","After three weeks, the degree of damage to the plants is rated in % damage in comparison to the development of the untreated control.","The figures denote: 0% =no effect (like untreated control) 100% =total destruction Post-emergence/ g.a.i./ Avena Greenhouse ha Alopecurus fatua Setaria Sinapis I-1-a-2 250 90 100 100 80 I-1-a-3 250 90 100 100 80 I-1-c-3 250 100 100 100 80 I-1-a-1 250 100 100 100 70 Post-emergence/ g.a.i/ Sugar- Alope- Avena Echino- Greenhouse ha beet curus fatua chloa Setaria I-2-a-1 250 0 90 95 100 100 Post-emergence/ g.a.i/ Sugar- Avena Echino- Greenhouse ha beet fatua chloa Setaria Sinapis I-1-c-1 250 0 90 100 100 70 Post-emergence/ g.a.i/ Alope- Avena Greenhouse ha curus fatua Setaria Galium I-2-c-1 250 90 100 100 80 pre-emergence/ Soya greenhouse g a.i./ha Wheat bean Digitaria Lolium Setaria Matricana I-1-a-2 60 0 0 100 100 100 95 pre-emergence/ Sugar Avena greenhouse g a.i./ha beet Alopercurus fatua Digitaria Polygonume I-1-a-3 125 0 95 100 100 100 pre-emergence/ g.a.i./ Oilseed Alope- Digi- greenhouse ha rape curus taria Lolium Stellaria I-1-c-2 125 0 95 100 100 100 pre-emergence/ g.a.i./ Soya- Alope- Seta- Stel-a greenhouse ha Wheat bean curus Lolium ria lari I-1-c-3 125 10 0 95 100 100 70 pre-emergence/ g.a.i./ Soya- greenhouse ha bean Lolium Setaria Matricaria Stellaria I-1-a-1 125 0 100 100 100 100 pre-emergence/ g.a.i./ Soya- greenhouse ha bean Bromus Lolium Setaria Abutilon I-2-a-1 125 0 95 100 100 70 Example G In vitro test for the ED50 determination in microorganisms A methanolic solution of the active compound to be tested, admixed with the emulsifier PS16, is pipetted into the wells of microtitre plates.","After the solvent has evaporated, 200 μl of potato/dextrose medium are added to each well.","Beforehand, a suitable concentration of spores or mycelium of the fungus to be tested was added to the medium.","The resulting concentrations of the active compound are 0.1, 1, 10 and 100 ppm.","The resulting concentration of the emulsifier is 300 ppm.","The plates are then incubated on a shaker at a temperature of 22° C. for 3-5 days, until sufficient growth can be observed in the untreated control.","Evaluation is carried out photometrically at a wavelength of 620 nm.","The dose of active compound which causes a 50% inhibition of fungal growth compared to the untreated control (ED50) is calculated from the data measured at different concentrations.","Active compound Microorganism ED50 value Ex.","I-1-a-2 Pyricularia oryzae <0.1 Septoria tritici <0.1 Ustilago avenae <0.1 Ex.","I-2-a-2 Pyricularia oryzae 1.96 Septoria tritici <0.1 Ustilago avenae <0.1 Ex.","I-2-a-3 Pyricularia oryzae <0.1 Septoria tritici <0.1 Ustilago avenae 0.1 Example H Sphaerotheca Test (Cucumber)/Protective Solvent: 24.5 parts by weight of acetone 24.5 parts by weight of dimethylacetamide Emulsifier: 1 part by weight of alkyl-aryl polyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amounts of solvent and emulsifier, and the concentrate is diluted with water to the desired concentration.","To test for protective activity, young plants are sprayed with the preparation of active compound at the stated application rate.","After the spray coating has dried on, the plants are inoculated with an aqueous spore suspension of Sphaerotheca fuliginea.","The plants are then placed in a greenhouse at about 23° C. and a relative atmospheric humidity of about 70%.","Evaluation is carried out 7 days after the inoculation.","0% means an efficacy which corresponds to that of the control, whereas an efficacy of 100% means that no infection is observed.","Application rate of active Active compound compound in g/ha % efficacy Ex.","I-1-a-2 100 100 Example I Venturia Test (Apple)/Protective Solvent: 24.5 parts by weight of acetone 24.5 parts by weight of dimethylacetamide Emulsifier: 1 part by weight of alkyl-aryl polyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amounts of solvent and emulsifier, and the concentrate is diluted with water to the desired concentration.","To test for protective activity, young plants are sprayed with the preparation of active compound at the stated application rate.","After the spray coating has dried on, the plants are inoculated with an aqueous conidia suspension of the apple scab pathogen Venturia inaequalis and then remain in an incubation cabin at about 20° C. and 100% relative atmospheric humidity for 1 day.","The plants are then placed in a greenhouse at about 21° C. and a relative atmospheric humidity of about 90%.","Evaluation is carried out 10 days after the inoculation.","0% means an efficacy which corresponds to that of the control, whereas an efficacy of 100% means that no infection is observed.","Application rate of active Active compound compound in g/ha % efficacy Ex.","I-1-a-2 100 91 Example J Alternaria Test (Tomato)/Protective Solvent: 24.5 parts by weight of acetone 24.5 parts by weight of dimethylacetamide Emulsifier: 1 part by weight of alkyl-arylpolyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amounts of solvent and emulsifier, and the concentrate is diluted with water to the desired concentration.","To test for protective activity, young plants are sprayed with the preparation of active compound at the stated application rate.","After the spray coating has dried on, the plants are inoculated with an aqueous spore suspension of Alternaria solani.","The plants are then placed in an incubation cabin at about 200° C. and 100% relative atmospheric humidity.","Evaluation is carried out 3 days after the inoculation.","0% means an efficacy which corresponds to that of the control, whereas an efficacy of 100% means that no infection is observed.","Application rate of active Active compound compound in g/ha % efficacy Ex.","I-1-a-2 100 93 Example K Botrytis Test (Bean)/Protective Solvent: 24.5 parts by weight of acetone 24.5 parts by weight of dimethylacetamide Emulsifier: 1 part by weight of alkyl-arylpolyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amounts of solvent and emulsifier, and the concentrate is diluted with water to the desired concentration.","To test for protective activity, young plants are sprayed with the preparation of active compound at the stated application rate.","After the spray coating has dried on, two small pieces of agar colonized by Botrytis cinerea are placed onto each leaf.","The inoculated plants are placed in a dark chamber at about 20° C. and 100% relative atmospheric humidity.","2 days after the inoculation, the size of the infested areas on the leaves is evaluated.","0% means an efficacy which corresponds to that of the control, whereas an efficacy of 100% means that no infection is observed.","Application rate of active Active compound compound in g/ha % efficacy Ex.","I-1-a-2 500 92 Example L Critical Concentration Test/Soil Insects-Treatment of Transgenic Plants Test insect: Diabrotica balteata—larvae in soil Solvent: 7 parts by weight of acetone Emulsifier: 1 part by weight of alkylaryl polyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amount of solvent, the stated amount of emulsifier is added and the concentrate is diluted with water to the desired concentration.","The preparation of active compound is poured onto the soil.","The concentration of the active compound in the preparation is virtually immaterial, only the amount by weight of active compound per volume unit of soil, which is stated in ppm (mg/l) matters.","The soil is filled into 0.25 l pots, and these are allowed to stand at 20° C. Immediately after the preparation, 5 pregerminated maize corns of the cultivar YIELD GUARD (trademark of Monsanto Comp., USA) are placed into each pot.","After 2 days, the corresponding test insects are placed into the treated soil.","After a further 7 days, the efficacy of the active compound is determined by counting the number of maize plants that emerged (1 plant=20% activity).","Example M Heliothis Virescens Test—Treatment of Transgenic Plants Solvent: 7 parts by weight of acetone Emulsifier: 1 part by weight of alkylaryl polyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amount of solvent and the stated amount of emulsifier, and the concentrate is diluted with water to the desired concentration.","Soya bean shoots (Glycine max) of the cultivar Roundup Ready (trademark of Monsanto Comp., USA) are treated by being dipped into the preparation of active compound of the desired concentration and are populated with the tobacco budworm caterpillar Heliothis virescens while the leaves are still moist.","After the desired period of time, the kill of the insects is determined."]],"string":"[\n [\n \"Thiazolyl-substsituted carbocylic 1,3-diones as pesticidal agents\",\n \"The present invention relates to novel hetaryl-substituted carbocyclic 1,3-diones of the formula (I) in which A, B, Q1, Q2, Q3, Q4, G, m and Het are as defined in the description, to a plurality of processes for their preparation and to their use as pesticides, herbicides and fungicides.\"\n ],\n [\n \"1.A compounds of the Formula (I) in which Het represents a nitrogen-containing 5-membered heterocycle, preferably selected from the group consisting of thiazolyl, which heterocycle is optionally substituted by halogen, alkyl, alkoxy, alkenyloxy, halogenoalkyl, halogenoalkoxy, halogenoalkenyloxy, cyano, nitro, alkylthio, alkylsulphinyl, alkylsulphonyl, optionally substituted phenyl or phenoxy, m represents the number 0 or 1, A represents hydrogen, represents in each case optionally halogen-substituted alkyl, alkenyl, alkoxyalkyl, polyalkoxyalkyl, alkylthioalkyl, saturated or unsaturated, optionally substituted cycloalkyl in which optionally at least one ring atom is replaced by a heteroatom, or in each case optionally halogen-, alkyl-, halogenoalkyl-, alkoxy-, halogenoalkoxy-, cyano- or nitro-substituted aryl, arylalkyl or hetaryl, B represents hydrogen or alkyl, A and B together with the carbon atom to which they are attached represent a saturated or unsaturated, unsubstituted or substituted cycle which optionally contains at least one heteroatom, A and Q1 together represent optionally substituted alkanediyl in which two not directly adjacent carbon atoms optionally form a further optionally substituted cycle, Q1 represents hydrogen, alkyl, alkoxyalkyl, optionally substituted cycloalkyl (in which optionally one methylene group is replaced by oxygen or sulphur) or in each case optionally substituted phenyl, hetaryl, phenylalkyl or hetarylalkyl, Q2, Q3, Q4 independently of one another represent hydrogen or alkyl, Q1 and Q2 together with the carbon atom to which they are attached represent a saturated or unsaturated, unsubstituted or substituted cycle which optionally contains one heteroatom, G represents hydrogen (a) or represents one of the groups in which E represents a metal ion or an ammonium ion, L represents oxygen or sulphur, M represents oxygen or sulphur, R1 represents in each case optionally halogen-substituted alkyl, alkenyl, alkoxyalkyl, alkylthioalkyl, polyalkoxyalkyl or optionally halogen-, alkyl- or alkoxy-substituted cycloalkyl which may be interrupted by at least one heteroatom, in each case optionally substituted phenyl, phenylalkyl, hetaryl, phenoxyalkyl or hetaryloxyalkyl, R2 represents in each case optionally halogen-substituted alkyl, alkenyl, alkoxyalkyl, polyalkoxyalkyl or represents in each case optionally substituted cycloalkyl, phenyl or benzyl, R3 represents in each case optionally substituted alkyl, halogenoalkyl, phenyl or benzyl, R4 and R5 independently of one another represent in each case optionally halogen-substituted alkyl, alkoxy, alkylamino, dialkylamino, alkylthio, alkenylthio, cycloalkylthio or represent in each case optionally substituted phenyl, benzyl, phenoxy or phenylthio, R6 and R7 independently of one another represent hydrogen, in each case optionally halogen-substituted alkyl, cycloalkyl, alkenyl, alkoxy, alkoxyalkyl, represent optionally substituted phenyl, represent optionally substituted benzyl, or together with the N atom to which they are attached represent a cycle which is optionally interrupted by oxygen or sulphur.\",\n \"2.A compounds of the formula (I) according to claim 1, in which Het represents m represents the number 0 or 1, X represents hydrogen, halogen, C1-C6-alkyl, C1-C6-alkoxy, C3-C6-alkenyloxy, nitro or cyano, Y represents halogen, C1-C6-alkyl, C1-C6-halogenoalkyl, C1-C6-alkoxy, C1-C6-halogenoalkoxy or represents the groups V1 represents hydrogen, halogen, C1-C12-alkyl, C1-C6-alkoxy, C1-C6-alkylthio, C1-C6-alkylsulphinyl, C1-C6-alkylsulphonyl, C1-C4-halogenoalkyl, C1-C4-halogenoalkoxy, nitro, cyano or represents phenyl, phenoxy, phenoxy-C1-C4-alkyl, phenyl-C1-C4-alkoxy, phenylthio-C1-C4-alkyl or phenyl-C1-C4-alkylthio, each of which is optionally mono- or polysubstituted by halogen, C1-C6-alkyl, C1-C6-alkoxy, C1-C4-halogenoalkyl, C1-C4-halogenoalkoxy, nitro or cyano, V2 and V3 independently of one another represent hydrogen, halogen, C1-C6-alkyl, C1-C6-alkoxy, C1-C4-halogenoalkyl or C1-C4-halogeno-alkoxy, V1 and V2 jointly together with the carbon atoms to which they are attached represent an optionally C1-C4-alkyl- or halogen-substituted 5- or 6-membered cycle in which optionally one or two carbon atoms may be replaced by oxygen, sulphur or nitrogen, A represents hydrogen or in each case optionally halogen-substituted C1-C12-alkyl, C3-C8-alkenyl, C1-C6-alkoxy-C1-C4-alkyl, optionally halogen-, C1-C4-alkyl- or C1-C4-alkoxy-substituted C3-C8-cycloalkyl or C3-C6-cycloalkyl-C1-C4-alkyl, in which optionally one or two not directly adjacent ring members are replaced by oxygen and/or sulphur, or represents in each case optionally halogen-, C1-C6-alkyl-, C1-C6-halogenoalkyl-, C1-C6-alkoxy-, C1-C6-halogenoalkoxy-, cyano- or nitro-substituted phenyl, benzyl, hetaryl having 5 or 6 ring atoms or hetaryl-C1-C4-alkyl having 5 or 6 ring atoms, B represents hydrogen or C1-C6-alkyl, A, B and the carbon atom to which they are attached represent saturated C3-C10-cycloalkyl or unsaturated C5-C10-cycloalkyl in which optionally one ring member is replaced by oxygen or sulphur and which are optionally mono- or disubstituted by C1-C6-alkyl, C3-C8-cycloalkyl, C1-C6-halogenoalkyl, C1-C6-alkoxy, C1-C6-alkylthio, halogen or phenyl, A and Q1 together represent C3-C6-alkanediyl which is optionally mono- or disubstituted by identical or different substituents from the group consisting of C1-C4-alkyl and C1-C4-alkoxy, Q1 represents hydrogen, C1-C6-alkyl, C1-C6-alkoxy-C1-C2-alkyl, optionally fluorine-, chlorine-, C1-C4-alkyl-, C1-C2-halogenoalkyl- or C1-C4-alkoxy-substituted C3-C8-cycloalkyl in which optionally one methylene group is replaced by oxygen or sulphur or in each case optionally halogen-, C1-C4-alkyl-, C1-C4-alkoxy-, C1-C2-halogenoalkyl-, C1-C2-halogenoalkoxy-, cyano- or nitro-substituted phenyl, pyridyl, thienyl, thiazolyl, phenyl-C1-C4-alkyl, pyridyl-C1-C2-alkyl or thiazolyl-C1-C2-alkyl, Q2, Q3, Q4 independently of one another represent hydrogen or C1-C4-alkyl, Q1 and Q2 together with the carbon atom to which they are attached represent optionally C1-C6-alkyl-, C1-C6-alkoxy- or C1-C2-halogenoalkyl-substituted C3-C7-cycloalkyl in which optionally one ring member is replaced by oxygen or sulphur, G represents hydrogen (a) or represents one of the groups in which E represents a metal ion or an ammonium ion, L represents oxygen or sulphur and M represents oxygen or sulphur, R1 represents in each case optionally halogen-substituted C1-C20-alkyl, C2-C20-alkenyl, C1-C8-alkoxy-C1-C8-alkyl, C1-C8-alkylthio-C1-C8-alkyl, poly-C1-C8-alkoxy-C1-C8-alkyl or optinally halogen-, C1-C6-alkyl- or C1-C6-alkoxy-substituted C3-C8-cycloalkyl, in which optionally one or more not directly adjacent ring members are replaced by oxygen and/or sulphur, represents optionally halogen-, cyano-, nitro-, C1-C6-alkyl, C1-C6-alkoxy-, C1-C6-halogenoalkyl-, C1-C6-halogenoalkoxy-, C1-C6-alkylthio or C1-C6-alkylsulphonyl-substituted phenyl, represents optionally halogen-, nitro-, cyano-, C1-C6-alkyl-, C1-C6-alkoxy-, C1-C6-halogenoalkyl- or C1-C6-halogenoalkoxy-substituted phenyl-C1-C6-alkyl, represents optionally halogen-, C1-C6-alkyl- or trifluoromethyl-substituted 5- or 6-membered hetaryl, represents optionally halogen- or C1-C6-alkyl-substituted phenoxy-C1-C6-alkyl or represents optionally halogen-, amino- or C1-C6-alkyl-substituted 5- or 6-membered hetaryloxy-C1-C6-alkyl, R2 represents in each case optionally halogen-substituted C1-C20-alkyl, C2-C20-alkenyl, C1-C8-alkoxy-C2-C8-alkyl, poly-C1-C8-alkoxy-C2-C8-alkyl, represents optionally halogen-, C1-C6-alkyl- or C1-C6-alkoxy-substituted C3-C8-cycloalkyl or represents in each case optionally halogen-, cyano-, nitro-, C1-C6-alkyl-, C1-C6-alkoxy-, C1-C6-halogenoalkyl- or C1-C6-halogenoalkoxy-substituted phenyl or benzyl, R3 represents optionally halogeno-substituted C1-C8-alkyl or represents in each case optionally halogen-, C1-C6-alkyl-, C1-C6-alkoxy-, C1-C4-halogenoalkyl-, C1-C4-halogenoalkoxy-, cyano- or nitro-substituted phenyl or benzyl, R4 and R5 independently of one another represent in each case optionally halogen-substituted C1-C8-alkyl, C1-C8-alkoxy, C1-C8-alkylamino, di-(C1-C8-alkyl)amino, C1-C8-alkylthio, C2-C8-alkenylthio, C3-C7-cycloalkylthio or represent in each case optionally halogen-, nitro-, cyano-, C1-C4-alkoxy-, C1-C4-halogenoalkoxy-, C1-C4-alkylthio-, C1-C4-halogenoalkylthio-, C1-C4-alkyl- or C1-C4-halogenoalkyl-substituted phenyl, benzyl, phenoxy or phenylthio, R6 and R7 independently of one another represent hydrogen, represent in each case optionally halogen-substituted C1-C8-alkyl, C3-C8-cycloalkyl, C1-C8-alkoxy, C3-C8-alkenyl, C1-C8-alkoxy-C1-C8-alkyl, represent optionally halogen-, C1-C8-halogenoalkyl-, C1-C8-alkyl- or C1-C8-alkoxy-substituted phenyl, optionally halogen-, C1-C8-alkyl-, C1-C8-halogenoalkyl- or C1-C8-alkoxy-substituted benzyl or together with the nitrogen to which they are attached represent an optionally C1-C4-alkyl-substituted C3-C6-alkylene radical in which optionally one carbon atom is replaced by oxygen or sulphur.\",\n \"3.A compound of the Formula (I) according to claim 1, in which Het represents m represents the number 0 or 1, X represents hydrogen, chlorine, bromine or C1-C4-alkyl, Y represents chlorine, bromine, C1-C4-alkyl, C1-C4-halogenoalkyl, C1-C4-alkoxy, C1-C4-halogenoalkoxy or the group V1 represents hydrogen, fluorine, chlorine, C1-C6-alkyl, C1-C4-alkoxy, C1-C2-halogenoalkyl, C1-C2-halogenoalkoxy, nitro, cyano or phenoxy which is optionally mono- or disubstituted by fluorine, chlorine, bromine, C1-C4-alkyl, C1-C4-alkoxy, C1-C2-halogenoalkyl, C1-C2-halogenoalkoxy, nitro or cyano, V2 represents hydrogen, fluorine, chlorine, bromine, C1-C4-alkyl, C1-C4-alkoxy, C1-C2-halogenoalkyl or C1-C2-halogenoalkoxy, V1 and V2 jointly, together with the carbon atoms to which they are attached, represent an optionally fluorine- or methyl-substituted 5- or 6-membered cycle in which optionally one or two carbon atoms may be replaced by oxygen, A represents hydrogen, represents C1-C8-alkyl or C1-C4-alkoxy-C1-C2-alkyl, each of which is optionally mono- to trisubstituted by fluorine, represents C5-C6-cycloalkyl or C3-C6-cycloalkyl-C1-C2-alkyl in which optionally one ring member is replaced by oxygen or sulphur, each of which radicals is optionally mono- or disubstituted by fluorine, chlorine, methyl, ethyl or methoxy, or represents phenyl or benzyl, each of which is optionally mono- or disubstituted by fluorine, chlorine, bromine, C1-C4-alkyl, C1-C2-halogenoalkyl, C1-C4-alkoxy or C1-C2-halogenoalkoxy, B represents hydrogen or C1-C4-alkyl, A, B and the carbon atom to which they are attached represent saturated C5-C6-cycloalkyl in which optionally one ring member is replaced by oxygen and which is optionally monosubstituted by C1-C4-alkyl, trifluoromethyl or C1-C4-alkoxy, with the proviso that in this case Q1 represents only hydrogen or C1-C4-alkyl, A and Q1 together represent C3-C4-alkanediyl which is optionally mono- or disubstituted by methyl, ethyl, methoxy or ethoxy, Q1 represents hydrogen, C1-C4-alkyl, C1-C4-alkoxy-C1-C2-alkyl, or optionally methyl- or methoxy-substituted C3-C6-cycloalkyl, in which optionally one methylene group is replaced by oxygen, or represents phenyl or benzyl, each of which is optionally mono- or disubstituted by fluorine, chlorine, bromine, C1-C4-alkyl, C1-C4-alkoxy, trifluoromethyl or trifluoromethoxy, Q2, Q3, Q4 independently of one another represent hydrogen, methyl or ethyl, Q1 and Q2 together with the carbon atom to which they are attached represent optionally C1-C4-alkyl- or C1-C4-alkoxy-substituted saturated C5-C6-cycloalkyl in which optionally one ring member is replaced by oxygen, with the proviso that in this case A only represents hydrogen or C1-C4-alkyl, G represents hydrogen (a) or represents one of the groups in which E represents a metal ion or an ammonium ion, L represents oxygen or sulphur and M represents oxygen or sulphur, R1 represents C1-C16-alkyl, C2-C16-alkenyl, C1-C4-alkoxy-C1-C2-alkyl, C1-C4-alkylthio-C1-C2-alkyl, each of which is optionally mono- to pentasubstituted by fluorine or chlorine, or C3-C6-cycloalkyl in which optionally one or two not directly adjacent ring members are replaced by oxygen and/or sulphur and which is optionally mono- or disubstituted by fluorine, chlorine, C1-C4-alkyl or C1-C5-alkoxy, represents phenyl which is optionally mono- or disubstituted by fluorine, chlorine, bromine, cyano, nitro, C1-C4-alkyl, C1-C4-alkoxy, trifluoromethyl or trifluoromethoxy, represents pyridyl or thienyl, each of which is optionally monosubstituted by fluorine, chlorine, bromine, methyl, ethyl or trifluoromethyl, R2 represents C1-C16-alkyl, C2-C16-alkenyl or C 1 -C4-alkoxy-C2-C4-alkyl, represents C3-C6-cycloalkyl which is optionally mono- or disubstituted by methyl, ethyl or methoxy, represents phenyl or benzyl, each of which is optionally mono- or disubstituted by fluorine, chlorine, bromine, cyano, nitro, C1-C4-alkyl, C1-C4-alkoxy, trifluoromethyl or trifluoromethoxy, R3 represents C1-C4-alkyl which is optionally mono- to pentasubstituted by fluorine or represents phenyl which is optionally mono- or disubstituted by fluorine, chlorine, bromine, C1-C4-alkyl, C1-C4-alkoxy, trifluoromethyl, trifluoromethoxy, cyano or nitro, R4 represents C1-C6-alkyl, C1-C6-alkoxy, C1-C6-alkylamino, di-(C1-C6-alkyl)amino, C1-C6-alkylthio, or represents phenyl, benzyl, phenoxy or phenylthio, each of which is optionally mono- or disubstituted by fluorine, chlorine, bromine, nitro, cyano, C1-C3-alkoxy, trifluoromethoxy, C1-C3-alkyl or trifluoromethyl, R5 represents C1-C4-alkyl, C1-C4-alkoxy or C1-C4-alkylthio, R6 represents hydrogen, C1-C6-alkyl, C3-C6-cycloalkyl, C1-C6-alkoxy, C3-C6-alkenyl, C1-C6-alkoxy-C1-C6-alkyl, represents phenyl which is optionally mono- or disubstituted by fluorine, chlorine, bromine, trifluoromethyl, C1-C4-alkyl or C1-C4-alkoxy, represents benzyl which is optionally substituted by fluorine, chlorine, bromine, methyl, ethyl, trifluoromethyl or methoxy, R7 represents hydrogen, C1-C6-alkyl or C3-C6-alkenyl, R6 and R7 together with nitrogen atom to which they are attached represent a C5-C6-alkylene radical in which optionally one methylene group is replaced by oxygen or sulphur and which is optionally mono- or disubstituted by methyl or ethyl.\",\n \"4.A compound of the Formula (I) according to claim 1, in which Het represents m represents the number 0 or 1, X represents hydrogen, chlorine, bromine, methyl, ethyl, n-propyl or isopropyl, Y represents the group V1 represents hydrogen, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, methoxy, ethoxy, n-propoxy, isopropoxy, trifluoromethyl, trifluoromethoxy or 4-chlorophenoxy, V2 represents hydrogen, fluorine, chlorine, bromine, methyl, methoxy or trifluoromethyl, A represents hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, methoxymethyl, ethoxymethyl, phenyl or cyclohexyl, B represents hydrogen, methyl or ethyl, A, B and the carbon atom to which they are attached represent saturated C5-C6-cycloalkyl in which optionally one ring member is replaced by oxygen and which is optionally monosubstituted by methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, trifluoromethyl, methoxy, ethoxy, n-propoxy, n-butoxy or isobutoxy, with the provisio that in this case Q1 only represents hydrogen, A and Q1 together represent C3-C4-alkanediyl, Q1 represents hydrogen, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, cyclopentyl, cyclohexyl or 4-chlorophenyl, Q2, Q3, Q4 independently of one another represent hydrogen, methyl or ethyl, Q1 and Q2 together with the carbon atom to which they are attached represent optionally methyl-, ethyl-, methoxy-, ethoxy-, n-propoxy- or n-butoxy-substituted saturated C5-C6-cycloalkyl in which optionalloy one ring member is replaced by oxygen, with the proviso that in this case A only represents hydrogen, G represents hydrogen (a) or represents one of the groups in which E represents a metal ion or an ammonium ion, L represents oxygen or sulphur and M represents oxygen or sulphur, R1 represents in each case optionally fluorine- or chlorine-substituted C1-C8-alkyl, C2-C8-alkenyl, C1-C2-alkoxy-C1-C2-alkyl, C1-C2-alkylthio-C1-C2-alkyl or cyclopropyl or cyclohexyl, each of which is optionally monosubstituted by fluorine, chlorine, methyl, ethyl or methoxy, represents phenyl which is optionally monosubstituted by fluorine, chlorine, bromine, cyano, nitro, methyl, ethyl, isopropyl, tert-butyl, methoxy, trifluoromethyl or trifluoromethoxy, represents thienyl or pyridyl, each of which is optionally monosubstituted by fluorine, chlorine, bromine or methyl, R2 represents C1-C8-alkyl, C2-C8-alkenyl or C1-C4-alkoxy-C2-alkyl, represents C3-C6-cycloalkyl which is optionally monosubstituted by methyl, ethyl or methoxy, represents phenyl or benzyl, each of which is optionally monosubstituted by fluorine, chlorine, cyano, nitro, methyl, ethyl, isopropyl, tert-butyl, methoxy, trifluoromethyl or trifluoromethoxy, R3 represents methyl or ethyl, each of which is optionally trisubstituted by fluorine or represents phenyl which is optionally monosubstituted by fluorine, chlorine, bromine, methyl, tert-butyl, methoxy, trifluoromethyl, trifluoromethoxy, cyano or nitro, R4 represents C1-C4-alkyl, C1-C4-alkoxy, C1-C4-alkylamino, di-(C1-C4-alkyl)amino, C1-C4-alkylthio or represents phenyl, phenoxy or phenylthio, each of which is optionally monosubstituted by fluorine, chlorine, bromine, nitro, cyano, C1-C2-alkoxy, trifluoromethoxy or C1-C3-alkyl, R5 represents methyl, ethyl, methoxy, ethoxy, methylthio or ethylthio, R6 represents hydrogen, represents C1-C4-alkyl, C3-C6-cycloalkyl, C1-C4-alkoxy, C3-C4-alkenyl or C1-C4-alkoxy-C1-C4-alkyl, R7 represents hydrogen, C1-C4-alkyl or C3-C4-alkenyl, R6 and R7 together with the nitrogen atom to which they are attached represent a C5-alkylene radical in which optionally one methylene group is replaced by oxygen.\",\n \"5.A compound of the Formula (I) according to claim 1, in which Het represents m represents the number 0 or 1, X represents hydrogen, methyl or ethyl, Y represents the group V1 represents hydrogen, chlorine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, trifluoromethyl or represents 4-chloro-phenoxy, A represents hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, phenyl or cyclohexyl, B represents hydrogen, methyl or ethyl, A, B and the carbon atom to which they are attached represent saturated C5-C6-cycloalkyl in which optionally one ring member is replaced by oxygen and which is optionally monosubstituted by methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, methoxy, ethoxy, n-propoxy, n-butoxy or isobutoxy, with the proviso that in this case Q1 only represents hydrogen, A and Q1 together represent C3-C4-alkanediyl, Q1 represents hydrogen, methyl, ethyl, n-propyl, isopropyl or 4-chlorophenyl, Q2 represents hydrogen, methyl or ethyl, Q3 represents hydrogen, Q4 represents hydrogen, Q1 and Q2 together with the carbon atom to which they are attached represent saturated C5-C6-cycloalkyl, with the proviso that in this case A only represents hydrogen, G represents hydrogen (a) or represents one of the groups in which L represents oxygen and M represents oxygen or sulphur, R1 represents C1-C8-alkyl, C1-C2-alkoxy-C1-C2-alkyl or cyclopropyl which is optionally monosubstituted by chlorine, represents phenyl which is optionally monosubstituted by chlorine, represents pyridyl which is optionally monosubstituted by chlorine, R2 represents C1-C8-alkyl, represents phenyl or benzyl, R3 represents methyl or ethyl, R6 and R7 together with the nitrogen atom to which they are attached represent a C5-alkylene radical in which optionally one methylene group is replaced by oxygen.\",\n \"6.A process for preparing a compound of the Formula (I) according to claim 1, wherein to obtain (A) a compound of the Formula (I-1-a) in which A, B, Q1, Q2 and Het are as defined in claim 1, a compound of the Formula (II) in which A, B, Q1, Q2 and Het are as defined in claim 1 and R8 represents alkyl, is condensed intramolecularly, optionally in the presence of a diluent and in the presence of a base, (B) a compound of the Formula (I-2-a) in which A, B, Q1, Q2, Q3, Q4, and Het are as defined claim 1, a compound of the Formula (III) in which A, B, Q1, Q2, Q3, Q4 and Het are as defined in claim 1 and R8 represents alkyl, is condensed intramolecularly in the presence of a diluent and in the presence of a base, (C) a compound of the Formulae (I-1-b) or (I-2-b) in which A, B, Q1, Q2, Q3, Q4, m, R1 and Het are as defined in claim 1, a compound of the Formulae (I-1-a) or (I-2-a) shown above in this claim 6 in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined in claim 1 are in each case (α) reacted with an acid halide of the Formula (IV) in which R1 is as defined in claim 1 and Hal represents halogen or (β) reacted with a carboxylic anhydride of the Formula (V) R1—CO—O—CO—R1 (V) in which R1 is as defined in claim 1, optionally in the presence of a diluent and optionally in the presence of an acid binder; (D) a compound of the Formulae (I-1-c) or (I-2-c) in which A, B, Q1 , Q2, Q3, Q4, m, R2, M and Het are as defined in claim 1 and L represents oxygen, compounds of the Formulae (I-1-a) or (I-2-a) shown above in this claim 6 in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined in claim 1 are in each case reacted with a chloroformic ester or a chloroformic thioester of the Formula (VI) R2—M—CO—Cl (VI) in which R2 and M are as defined in claim 1, optionally in the presence of a diluent and optionally in the presence of an acid binder; (E) a compound of the Formulae (I-1-c) or (I-2-c) shown above in this claim 6 in which A, B, Q1, Q2, Q3, Q4, m, R2, M and Het are as defined in claim 1 and L represents sulphur, compounds of the Formulae (I-1 -a) or (I-2-a) shown above in this claim 6 in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined in claim 1 are in each case reacted with a chloromonothioformic ester or a chlorodithioformic ester of the Formula (VII) in which M and R2 are as defined in claim 1, optionally in the presence of a diluent and optionally in the presence of an acid binder and (F) a compound of the Formulae (I-1-d) or (I-2-d) in which A, B, Q1, Q2, Q3, Q4, m, R3 and Het are as defined in claim 1, compounds of the Formulae (I-1-a) or (I-2-a) shown above in this claim 6 which A, B, Q1, Q2, Q3, Q4, m and Het are as defined in claim 1 are in each case reacted with a sulphonyl chloride of the Formula (VIII) R3—SO2—Cl (VIII) in which R3 is as defined in claim 1, optionally in the presence of a diluent and optionally in the presence of an acid binder, (G) a compound of the Formulae (I-1-e) or (I-2-e) in which A, B, L, Q1, Q2, Q3, Q4, m, R4, R5 and Het are as defined in claim 1, compounds of the Formulae (I-1-a) or (I-2-a) shown above in this claim 6 in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined in claim 1 are in each case reacted with a phosphorus compound of the Formula (IX) in which L, R4 and R5 are as defined in claim 1 and Hal represents halogen, optionally in the presence of a diluent and optionally in the presence of an acid binder, (H) a compound of the formulae (I-1-f) or (I-2-f) in which A, B, E, Q1, Q2, Q3, Q4, m and Het are as defined in claim 1, compounds of the Formulae (I-1-a) or (I-2-a) shown above in this claim 6 in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined in claim 1 are in each case reacted with a metal compound or an amine of the Formulae (X) and (XI), respectively, Me(OR11)t (X) in which Me represents a mono- or divalent metal, t represents the number 1 or 2 and R11, R12, R13 independently of one another represent hydrogen or alkyl, optionally in the presence of a diluent, (I) a compound of the Formulae (I-1-g) or (I-2-g) in which A, B, L, Q1, Q2, Q3, Q4, m, R6, R7 and Het are as defined in claim 1, compounds of the Formulae (I-1-a) or (I-2-a) shown above in this claim 6 in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined in claim 1 are in each case (α) reacted with an isocyanater or isothiocyanater of the Formula (XII) R6—N═C═L (XII) in which R6 and L are as defined in claim 1, optionally in the presence of a diluent and optionally in the presence of a catalyst, or (β) reacted with a carbamoyl chloride or a thiocarbamoyl chloride of the Formula (XIII) in which L, R6 and R7 are as defined in claim 1, optionally in the presence of a diluent and optionally in the presence of an acid binder.\",\n \"7.A compound of the Formula (II) in which A, B, Q1, Q2 and Het are as defined in claim 1 and R8 represents alkyl.\",\n \"8.A compound of the Formula (III) in which A, B, Q1, Q2, Q3, Q4, and Het are as defined in claim 1, and R8 represents alkyl.\",\n \"9.A compound of the Formula (XIV) in which A, B, Het, Q1 and Q2 are as defined in claim 1.10.A compound of the Formula (XV) in which A, B, Het, Q1 and Q2 are as defined claim 1 and R8 and R8′ represents alkyl.\",\n \"11.A compound of the Formula (XIX) in which A, B, Het, Q1, Q2, Q3 and Q4 are as defined claim 1.12.A compound of the Formula (XX) in which A, B, Het, Q1, Q2, Q3, and Q4are as defined in claim 1, R8 and R8 represent alkyl.\",\n \"13.Pesticides, herbicides and fungicides comprising at least one compound of the Formula (I) according to claim 1.14.A method for controlling animal pests, undesirable vegetation and/or fungi comprising allowing a compound of the Formula (I) according to claim 1 to act on pests, undesirable vegetation and/or fungi and/or their habitat.\",\n \"15.canceled 16.A process for preparing pesticides, herbicides and/or fungicides comprising mixing one or more compounds of the Formula (I) according to claim 1 with one or more extenders and/or surfactants.\",\n \"17.canceled.\"\n ],\n [\n \"The present invention relates to novel hetaryl-substituted cyclopentane- and cyclohexane-1,3-dione derivatives, to a plurality of processes for their preparation and to their use as bioactive compounds in agriculture.\",\n \"2-(2-Amino-thiazol-4-yl)-3-hydroxy-5,5-dimethyl-cyclohex-2-enone is known; Pshenichnyi, V. N. et al; CNClALi; Chem.\",\n \"Heterocycl.\",\n \"Compd.\",\n \"(Engl.\",\n \"transl.)\",\n \"EN; 26, 1175-1178 (1990).\",\n \"This invention now provides novel compounds of the formula (I) in which Het represents a nitrogen-containing 5-membered heterocycle, preferably from the group consisting of thiazolyl, which heterocycle is optionally substituted by halogen, alkyl, alkoxy, alkenyloxy, halogenoalkyl, halogenoalkoxy, halogenoalkenyloxy, cyano, nitro, alkylthio, alkylsulphinyl, alkylsulphonyl, optionally substituted phenyl or phenoxy, m represents the number 0 or 1, A represents hydrogen, represents in each case optionally halogen-substituted alkyl, alkenyl, alkoxyalkyl, polyalkoxyalkyl, alkylthioalkyl, saturated or unsaturated, optionally substituted cycloalkyl in which optionally at least one ring atom is replaced by a heteroatom, or in each case optionally halogen-, alkyl-, halogenoalkyl-, alkoxy-, halogenoalkoxy-, cyano- or nitro-substituted aryl, arylalkyl or hetaryl, B represents hydrogen or alkyl, A and B together with the carbon atom to which they are attached represent a saturated or unsaturated, unsubstituted or substituted cycle which optionally contains at least one heteroatom, A and Q1 together represent optionally substituted alkanediyl in which two not directly adjacent carbon atoms optionally form a further optionally substituted cycle, Q1 represents hydrogen, alkyl, alkoxyalkyl, optionally substituted cycloalkyl (in which optionally one methylene group is replaced by oxygen or sulphur) or in each case optionally substituted phenyl, hetaryl, phenylalkyl or hetarylalkyl, Q2, Q3, Q4 independently of one another represent hydrogen or alkyl, Q1 and Q2 together with the carbon atom to which they are attached represent a saturated or unsaturated, unsubstituted or substituted cycle which optionally contains one heteroatom, G represents hydrogen (a) or represents one of the groups in which E represents a metal ion or an ammonium ion, L represents oxygen or sulphur, M represents oxygen or sulphur, R1 represents in each case optionally halogen-substituted alkyl, alkenyl, alkoxyalkyl, alkylthioalkyl, polyalkoxyalkyl or optionally halogen-, alkyl- or alkoxy-substituted cycloalkyl which may be interrupted by at least one heteroatom, in each case optionally substituted phenyl, phenylalkyl, hetaryl, phenoxyalkyl or hetaryloxyalkyl, R2 represents in each case optionally halogen-substituted alkyl, alkenyl, alkoxyalkyl, polyalkoxyalkyl or represents in each case optionally substituted cycloalkyl, phenyl or benzyl, R3 represents in each case optionally substituted alkyl, halogenoalkyl, phenyl or benzyl, R4 and R5 independently of one another represent in each case optionally halogen-substituted alkyl, alkoxy, alkylamino, dialkylamino, alkylthio, alkenylthio, cycloalkylthio or represent in each case optionally substituted phenyl, benzyl, phenoxy or phenylthio, R6 and R7 independently of one another represent hydrogen, in each case optionally halogen-substituted alkyl, cycloalkyl, alkenyl, alkoxy, alkoxyalkyl, represent optionally substituted phenyl, represent optionally substituted benzyl, or together with the N atom to which they are attached represent a cycle which is optionally interrupted by oxygen or sulphur.\",\n \"Depending inter alia on the nature of the substituents, the compounds of the formula (I) can be present as geometrical and/or optical isomers or isomer mixtures of varying composition which, if desired, can be separated in a known manner.\",\n \"The present invention provides both the pure isomers and the isomer mixtures, their preparation and use and the compositions comprising them.\",\n \"However, hereinbelow, for the sake of simplicity, only compounds of the formula (I) are referred to, although this includes both the pure compounds and, if appropriate, mixtures having varying proportions of isomeric compounds.\",\n \"Depending on the position of the substituent G, the compounds of the formula (I) can be present in the two isomeric forms of the formulae (I-α) and (I-β), which is meant to be expressed by the broken line in the formula (I).\",\n \"The compounds of the formulae (I-α) and (I-β) can be present either as mixtures or else in the form of their pure isomers.\",\n \"Mixtures of the compounds of the formulae (I-α) and (I-β) can, if desired, be separated in a manner known per se by physical methods, for example by chromatographic methods.\",\n \"For reasons of clarity, in each case only one of the possible isomers is shown hereinbelow.\",\n \"This does not preclude that the compounds are, if appropriate, present in the form of the isomer mixtures or in the respective other isomeric form.\",\n \"Including Het for thiazolyl and m for the numbers 0 and 1, the following principal structures (I-1) and (I-2) result Including the different meanings (a), (b), (c), (d,(e), (f) and (g) of the group G. the following principal structures (I-1-a) to (I-1-g) result if m represents the number 0 (I-1), in which A, B, E, L, M, Q1, Q2, R1, R2, R3, R4, R5, R6 and R7 are as defined above, and x represents hydrogen, halogen, alkyl, alkoxy, alkenyloxy, nitro, cyano or optionally substituted phenyl and Y represents halogen, alky, halogenoalkyl, alkoxy, halogenoalkoxy or in each case optionally substituted phenyl or phenoxy.\",\n \"Including the different meanings (a), (b), (c), (d), (e), (f) and (g) of the group G, the following principal structures (I-2-a) to (I-2-g) result if m represents the number 1 (I-2), in which A, B, E, L, M, Q1, Q2, Q3, Q4, X, Y, R1, R2, R3, R4, R5, R6 and R7 are as defined above.\",\n \"Furthermore, it has been found that the novel compounds of the formula (I) are obtained by one of the processes described below: (A) substituted cyclopentane-1,3-diones or their enoles of the formula (I-1-a) in which A, B, Q1, Q2 and Het are as defined above, are obtained when ketocarboxylic esters of the formula (II) in which A, B, Q1, Q2 and Het are as defined above and R8 represents alkyl (preferably C1-C6-alkyl) are condensed intramolecularly, if appropriate in the presence of a diluent and in the presence of a base.\",\n \"Furthermore, it has been found (B) that substituted cyclohexane-1,3-diones of the formula (1-2-a) in which A, B, Q1, Q2, Q3, Q4, and Het are as defined above are obtained when ketocarboxylic esters of the formula (III) in which A, B. Q, Q2, Q3, Q4, and Het are as defined above, and R8 represents alkyl (preferably C1-C6-alkyl) are condensed intramolecularly in the presence of a diluent and in the presence of a base.\",\n \"Furthermnore, it has been found (C) that the compounds of the formulae (I-1-b) to (I-2-b) shown above in which A, B, Q1, Q2, Q3, Q4, m, R1 and Het are as defined above are obtained when compounds of the formulae (I-1-a) to (I-2-a) shown above in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined above are in each case (α) reacted with acid halides of the formula (IV) in which R1 is as defined above and Hal represents halogen (in particular chlorine or bromine) or (β) reacted with carboxylic anhydrides of the formula (V) R1—CO—O—CO—R1 (V) in which R1 is as defined above, if appropriate in the presence of a diluent and if appropriate in the presence of an acid binder; (D) that the compounds of the formulae (I-1-c) to (I-2-c) shown above in which A, B, Q1, Q2, Q3, Q4, m, R2, M and Het are as defined above and L represents oxygen are obtained when compounds of the formulae (I-1-a) to (I-2-a) shown above in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined above are in each case reacted with chloroformic esters or chloroformic thioesters of the formula (VI) R2—M—CO—Cl (VI) in which R2 and M are as defined above, if appropriate in the presence of a diluent and if appropriate in the presence of an acid binder; (E) that compounds of the formulae (I-1-c) to (I-2-c) shown above in which A, B, Q1, Q2, Q3, Q4, m, R2, M and Het are as defined above and L represents sulphur are obtained when compounds of the formulae (I-1-a) to (I-2-a) shown above in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined above are in each case reacted with chloromonothioformic esters or chlorodithioformic esters of the formula (VII) in which M and R2 are as defined above, if appropriate in the presence of a diluent and if appropriate in the presence of an acid binder and (F) that compounds of the formulae (I-1-d) to (I-2-d) shown above in which A, B, Q1, Q2, Q3, Q4, m, R3 and Het are as defined above are obtained when compounds of the formulae (I-1-a) and (I-2-a) shown above in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined above are in each case reacted with sulphonyl chlorides of the formula (VIII) R3—SO2—Cl (VIII) in which R3 is as defined above, if appropriate in the presence of a diluent and if appropriate in the presence of an acid binder, (G) that compounds of the formulae (I-1-e) to (I-2-e) shown above in which A, B, L, Q1, Q2, Q3, Q4, m, R4, R5 and Het are as defined above are obtained when compounds of the formulae (I-1-a) to (I-2-a) shown above in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined above are in each case reacted with phosphorus compounds of the formula (IX) in which L, R4 and R5 are as defined above and Hal represents halogen (in particular chlorine or bromine), if appropriate in the presence of a diluent and if appropriate in the presence of an acid binder, (H) that compounds of the formulae (I-1-f) to (I-2-f) shown above in which A, B, E, Q1, Q2, Q3, Q4, m and Het are as defined above are obtained when compounds of the formulae (I-1-a) to (I-2-a) in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined above are in each case reacted with metal compounds or amines of the formulae (X) and (XI), respectively, in which Me represents a mono- or divalent metal (preferably an alkali metal or alkaline earth metal such as lithium, sodium, potassium, magnesium or calcium), t represents the number 1 or 2 and R11, R12, R13 independently of one another represent hydrogen or alkyl (preferably C1-C8-alkyl), if appropriate in the presence of a diluent, (I) that compounds of the formulae (I-1-g) to (I-2-g) shown above in which A, B, L, Q1, Q2, Q3, Q4, Q4, m, R6, R7 and Het are as defined above are obtained when compounds of the formulae (I-1-a) to (I-2-a) shown above in which A, B, Q1, Q2, Q3, Q4, m and Het are as defined above are in each case (α) reacted with isocyanates or isothiocyanates of the formula (XII) R6—N═C═L (XII) in which R6 and L are as defined above, if appropriate in the presence of a diluent and if appropriate in the presence of a catalyst, or (β) reacted with carbamoyl chlorides or thiocarbamoyl chlorides of the formula (XII) in which L, R6 and R7 are as defined above, if appropriate in the presence of a diluent and if appropriate in the presence of an acid binder.\",\n \"Furthermore, it has been found that the novel compounds of the formula (I) have, depending on the substituents, very good activity both as pesticides, preferably as insecticides and acaricides, and as herbicides and fungicides.\",\n \"The formula (I) provides a general definition of the compounds according to the invention.\",\n \"Preferred substituents or ranges of the radicals given in the formulae mentioned above and below are illustrated below: Het preferably represents m preferably represents the number 0 or 1, X preferably represents hydrogen, halogen, C1-C6-alkyl, C1-C6-alkoxy, C3-C6-alkenyloxy, nitro or cyano, Y preferably represents halogen, C1-C6-alkyl, C1-C6-halogenoalkyl, C1-C6-alkoxy, C1-C6-halogenoalkoxy or represents the groups V1 preferably represents hydrogen, halogen, C1-C12-alkyl, C1-C6-alkoxy, C1-C6-alkylthio, C1-C6-alkylsulphinyl, C1-C6-alkylsulphonyl, C1-C4-halogenoalkyl, C1-C4-halogenoalkoxy, nitro, cyano or represents phenyl, phenoxy, phenoxy-C1-C4-alkyl, phenyl-C1-C4-alkoxy, phenylthio-C1-C4-alkyl or phenyl-C1-C4-alkylthio, each of which is optionally mono- or polysubstituted by halogen, C1-C6-alkyl, C1-C6-alkoxy, C1-C4-halogenoalkyl, C1-C4-halogenoalkoxy, nitro or cyano, V2 and V3 independently of one another preferably represent hydrogen, halogen, C1-C6-alkyl, C1-C6-alkoxy, C1-C4-halogenoalkyl or C1-C4-halogenoalkoxy, V1 and V2 jointly together with the carbon atoms to which they are attached preferably represent an optionally C1-C4-alkyl- or halogen-substituted 5- or 6-membered cycle in which optionally one or two carbon atoms may be replaced by oxygen, sulphur or nitrogen, A preferably represents hydrogen or in each case optionally halogen-substituted C1-C12-alkyl, C3-C8-alkenyl, C1-C6-alkoxy-C1-C4-alkyl, optionally halogen-, C1-C4-alkyl- or C1-C4-alkoxy-substituted C3-C8-cycloalkyl or C3-C6-cycloalkyl-C1-C4-alkyl, in which optionally one or two not directly adjacent ring members are replaced by oxygen and/or sulphur, or represents in each case optionally halogen-, C1-C6-alkyl-, C1-C6-halogenoalkyl-, C1-C6-alkoxy-, C1-C6-halogenoalkoxy-, cyano- or nitro-substituted phenyl, benzyl, hetaryl having 5 or 6 ring atoms (for example furanyl, pyridyl, imidazolyl, triazolyl, pyrazolyl, pyrimidyl, thiazolyl or thienyl) or hetaryl-C1-C4-alkyl having 5 or 6 ring atoms (for example pyridyl-, pyrimidyl- or thiazolyl-C1-C4-alkyl), B preferably represents hydrogen or C1-C6-alkyl, A, B and the carbon atom to which they are attached preferably represent saturated C3-C11-cycloalkyl or unsaturated C5-C10-cycloalkyl in which optionally one ring member is replaced by oxygen or sulphur and which are optionally mono- or disubstituted by C1-C6-alkyl, C3-C8-cycloalkyl, C1-C6-halogenoalkyl, C1-C6-alkoxy, C1-C6-alkylthio, halogen or phenyl, A and Q1 together preferably represent C3-C6-alkanediyl which is optionally mono- or disubstituted by identical or different substituents from the group consisting of C1-C4-alkyl and C1-C4-alkoxy, Q1 preferably represents hydrogen, C1-C6-alkyl, C1-C6-alkoxy-C1-C2-alkyl, optionally fluorine-, chlorine-, C1-C4-alkyl-, C1-C2-halogenoalkyl- or C1-C4-alkoxy-substituted C3-C8-cycloalkyl in which optionally one methylene group is replaced by oxygen or sulphur or in each case optionally halogen-, C1-C4-alkyl-, C1-C4-alkoxy-, C1-C2-halogenoalkyl-, C1-C2-halogenoalkoxy-, cyano- or nitro-substituted phenyl, pyridyl, thienyl, thiazolyl, phenyl-C1-C4-alkyl, pyridyl -C11-C2-alkyl or thiazolyl-C1-C2-alkyl, Q2, Q3, Q4 independently of one another preferably represent hydrogen or C1-C4-alkyl, Q1 and Q2 together with the carbon atom to which they are attached preferably represent optionally C1-C6-alkyl-, C1-C6-alkoxy- or C1-C2-halogenoalkyl-substituted C3-C7-cycloalkyl in which optionally one ring member is replaced by oxygen or sulphur, G preferably represents hydrogen (a) or represents one of the groups in particular (a), (b), (c) or (g), in which E represents a metal ion or an ammonium ion, L represents oxygen or sulphur and M represents oxygen or sulphur, R1 preferably represents in each case optionally halogen-substituted C1-C20-alkyl, C2-C20-alkenyl, C1-C8-alkoxy-C1-C8-alkyl, C1-C8-alkylthio-C1-C8-alkyl, poly-C1-C8-alkoxy-C1-C8-alkyl or optionally halogen-, C1-C6-alkyl- or C1-C6-alkoxy-substituted C3-C8-cycloalkyl, in which optionally one or more (preferably one or two) not directly adjacent ring members are replaced by oxygen and/or sulphur, represents optionally halogen-, cyano-, nitro-, C1-C6-alkyl, C1-C6-alkoxy-, C1-C6-halogenoalkyl-, C1-C6-halogenoalkoxy-, C1-C6-alkylthio or C1-C6-alkylsulphonyl-substituted phenyl, represents optionally halogen-, nitro-, cyano-, C1-C6-alkyl-, C1-C6-alkoxy-, C1-C6-halogenoalkyl- or C1-C6-halogenoalkoxy-substituted phenyl-C1-C6-alkyl, represents optionally halogen-, C1-C6-alkyl- or trifluoromethyl-substituted 5- or 6-membered hetaryl (for example pyrazolyl, thiazolyl, pyridyl, pyrimidyl, furanyl or thienyl), represents optionally halogen- or C1-C6-alkyl-substituted phenoxy-C1-C6-alkyl or represents optionally halogen-, amino- or C1-C6-alkyl-substituted 5- or 6-membered hetaryloxy-C1-C6-alkyl (for example pyridyloxy-C1-C6-alkyl, pyrimidyloxy-C1-C6-alkyl or thiazolyloxy- C1-C6-alkyl), R2 preferably represents in each case optionally halogen-substituted C1-C20-alkyl, C2-C20-alkenyl, C1-C8-alkoxy-C2-C8-alkyl, poly-C1-C8-alkoxy-C2-C8-alkyl, represents optionally halogen-, C1-C6-alkyl- or C1-C6-alkoxy-substituted C3-C8-cycloalkyl or represents in each case optionally halogen-, cyano-, nitro-, C1-C6-alkyl-, C1-C6-alkoxy-, C1-C6-halogenoalkyl- or C1-C6-halogenoalkoxy-substituted phenyl or benzyl, R3 preferably represents optionally halogeno-substituted C1-C8-alkyl or represents in each case optionally halogen-, C1-C6-alkyl-, C1-C6-alkoxy-, C1-C4-halogenoalkyl-, C1-C4-halogenoalkoxy-, cyano- or nitro-substituted phenyl or benzyl, R4 and R5 independently of one another represent in each case optionally halogen-substituted C1-C8-alkyl, C1-C8-alkoxy, C1-C8-alkylamino, di-(C1-C8-alkyl)amino, C1-C8-alkylthio, C2-C8-alkenylthio, C3-C7-cycloalkylthio or represent in each case optionally halogen-, nitro-, cyano-, C1-C4-alkoxy-, C1-C4-halogenoalkoxy-, C1-C4-alkylthio-, C1-C4-halogenoalkylthio-, C1-C4-alkyl- or C1-C4-halogenoalkyl-substituted phenyl, benzyl, phenoxy or phenylthio, R6 and R7 independently of one another preferably represent hydrogen, represent in each case optionally halogen-substituted C1-C8-alkyl, C3-C8-cycloalkyl, C1-C8-alkoxy, C3-C8-alkenyl, C1-C8-alkoxy-C1-C8-alkyl, represent optionally halogen-, C1-C8-halogenoalkyl-, C1-C8-alkyl- or C1-C8-alkoxy-substituted phenyl, optionally halogen-, C1-C8-alkyl-, C1-C8-halogenoalkyl- or C1-C8-alkoxy-substituted benzyl or together with the nitrogen to which they are attached represent an optionally C1-C4-alkyl-substituted C3-C6-alkylene radical in which optionally one carbon atom is replaced by oxygen or sulphur.\",\n \"In the radical definitions mentioned as being preferred, halogen represents fluorine, chlorine, bromine and iodine, in particular fluorine and chlorine, also as substituent, such as, for example, in halogenoalkyl.\",\n \"Het particularly preferably represents m particularly preferably represents the number 0 or 1, X particularly preferably represents hydrogen, chlorine, bromine or C1-C4-alkyl, Y particularly preferably represents chlorine, bromine, C1-C4-alkyl, C1-C4-halogenoalkyl, C1-C4-alkoxy, C1-C4-halogenoalkoxy or the group V1 particularly preferably-represents hydrogen, fluorine, chlorine, C1-C6-alkyl, C1-C4-alkoxy, C1-C2-halogenoalkyl, C1-C2-halogenoalkoxy, nitro, cyano or phenoxy which is optionally mono- or disubstituted by fluorine, chlorine, bromine, C1-C4-alkyl, C1-C4-alkoxy, C1-C2-halogenoalkyl, C1-C2-halogenoalkoxy, nitro or cyano, V2 particularly preferably represents hydrogen, fluorine, chlorine, bromine, C1-C4-alkyl, C1-C4-alkoxy, C1-C2-halogenoalkyl or C1-C2-halogenoalkoxy, V1 and V2 jointly, together with the carbon atoms to which they are attached, particularly preferably represent an optionally fluorine- or methyl-substituted 5- or 6-membered cycle in which optionally one or two carbon atoms may be replaced by oxygen, A particularly preferably represents hydrogen, represents C1-C8-alkyl or C1-C4-alkoxy-C1-C2-alkyl, each of which is optionally mono- to trisubstituted fluorine, represents C5-C6-cycloalkyl or C3-C6-cycloalkyl-C1-C2-alkyl in which optionally one ring member is replaced by oxygen or sulphur, each of which radicals is optionally mono- or disubstituted by fluorine, chlorine, methyl, ethyl or methoxy, or represent phenyl or benzyl, each of which is optionally mono- or disubstituted by fluorine, chlorine, bromine, C1-C4-alkyl, C1-C2-halogenoalkyl, C1-C4-alkoxy or C1-C2-halogenoalkoxy, B particularly preferably represents hydrogen or C1-C4-alkyl, A, B and the carbon atom to which they are attached particularly preferably represent saturated C5-C6-cycloalkyl in which optionally one ring member is replaced by oxygen and which is optionally monosubstituted by C1-C4-alkyl, trifluoromethyl or C1-C4-alkoxy, with the proviso that in this case Q1 represents only hydrogen or C1-C4-alkyl, A and Q1 together particularly preferably represent C3-C4-alkanediyl which is optionally mono- or disubstituted by methyl, ethyl, methoxy or ethoxy, Q1 particularly preferably represents hydrogen, C1-C4-alkyl, C1-C4-alkoxy-C1-C2-alkyl, or optionally methyl- or methoxy-substituted C3-C6-cycloalkyl, in which optionally one methylene group is replaced by oxygen, or phenyl or benzyl, each of which is optionally mono- or disubstituted by fluorine, chlorine, bromine, C1-C4-alkyl, C1-C4-alkoxy, trifluoromethyl or trifluoromethoxy, Q2, Q3, Q4 independently of one another particularly preferably represent hydrogen, methyl or ethyl, Q1 and Q2 together with the carbon to which they are attached particularly preferably represent optionally C1-C4-alkyl- or C1-C4-alkoxy-substituted saturated C5-C6-cycloalkyl in which optionally one ring member is replaced by oxygen, with the proviso that in this case A only represents hydrogen or C1-C4-alkyl, G particularly preferably represents hydrogen (a) or represents one of the groups (g), in particular (a), (b) or (c) in which E represents a metal ion or an ammonium ion, L represents oxygen or sulphur and M represents oxygen or sulphur, R1 particularly preferably represents C1-C16-alkyl, C2-C16-alkenyl, C1-C4-alkoxy-C1-C2-alkyl, C1-C4-alkylthio-C1-C2-alkyl, each of which is optionally mono- to pentasubstituted by fluorine or chlorine, or C3-C6-cycloalkyl in which optionally one or two not directly adjacent ring members are replaced by oxygen and/or sulphur and which is optionally mono- or di substituted by fluorine, chlorine, C1-C4-alkyl or C1-C5-alkoxy, particularly preferably represents phenyl which is optionally mono- or disubstituted by fluorine, chlorine, bromine, cyano, nitro, C1-C4-alkyl, C1-C4-alkoxy, trifluoromethyl or trifluoromethoxy, particularly preferably represents pyridyl or thienyl, each of which is optionally monosubstituted by fluorine, chlorine, bromine, methyl, ethyl or trifluoromethyl, R2 particularly preferably represents C1-C16-alkyl, C2-C16-alkenyl or C1-C4-alkoxy-C2-C4-alkyl, particularly preferably represents C3-C6-cycloalkyl which is optionally mono- or disubstituted by methyl, ethyl or methoxy, particularly preferably represents phenyl or benzyl, each of which is optionally mono- or disubstituted by fluorine, chlorine, bromine, cyano, nitro, C1-C4-alkyl, C1-C4-alkoxy, trifluoromethyl or trifluoromethoxy, R3 particularly preferably represents C1-C4-alkyl which is optionally mono- to pentasubstituted by fluorine or represents phenyl which is optionally mono- or disubstituted by fluorine, chlorine, bromine, C1-C4-alkyl, C1-C4-alkoxy, trifluoromethyl, trifluoromethoxy, cyano or nitro, R4 particularly preferably represents C1-C6-alkyl, C1-C6-alkoxy, C1-C6-alkylamino, di-(C1-C6-alkyl)amino, C1-C6-alkylthio, or represents phenyl, benzyl, phenoxy or phenylthio, each of which is optionally mono- or disubstituted by fluorine, chlorine, bromine, nitro, cyano, C1-C3-alkoxy, trifluoromethoxy, C1-C3-alkyl or trifluoromethyl, R5 particularly preferably represents C1-C4-alkyl, C1-C4-alkoxy or C1-C4-alkylthio, R6 particularly preferably represents hydrogen, C1-C6-alkyl, C3-C6-cycloalkyl, C1-C6-alkoxy, C3-C6-alkenyl, C1-C6-alkoxy-C1-C6-alkyl, represents phenyl which is optionally mono- or disubstituted by fluorine, chlorine, bromine, trifluoromethyl, C1-C4-alkyl or C1-C4-alkoxy, represents benzyl which is optionally substituted by fluorine, chlorine, bromine, methyl, ethyl, trifluoromethyl or methoxy, R7 particularly preferably represents hydrogen, C1-C6-alkyl or C3-C6-alkenyl, R6 and R7 together, with nitrogen atom to which they are attached particularly preferably represent a C5-C6-alkylene radical in which optionally one methylene group is replaced by oxygen or sulphur and which is optionally mono- or disubstituted by methyl or ethyl.\",\n \"In the radical definitions mentioned as being particularly preferred, halogen represents fluorine, chlorine and bromine, in particular fluorine and chlorine, especially fluorine, including as substituent, such as, for example, in halogenoalkyl.\",\n \"Het very particularly preferably represents m very particularly preferably represents the number 0 or 1, X very particularly preferably represents hydrogen, chlorine, bromine, methyl, ethyl, n-propyl or isopropyl, Y very particularly preferably represents the group V1 very particularly preferably represents hydrogen, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, methoxy, ethoxy, n-propoxy, isopropoxy, trifluoromethyl, trifluoromethoxy or 4-chlorophenoxy, V2 very particularly preferably represents hydrogen, fluorine, chlorine, bromine, methyl, methoxy or trifluoromethyl, A very particularly preferably represents hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, methoxymethyl, ethoxymethyl, phenyl or cyclohexyl, B very particularly preferably represents hydrogen, methyl or ethyl, A, B and the carbon atom to which they are attached very particularly preferably represent saturated C5-C6-cycloalkyl in which optionally one ring member is replaced by oxygen and which is optionally monosubstituted by methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, trifluoromethyl, methoxy, ethoxy, n-propoxy, n-butoxy or isobutoxy, with the proviso that in this case Q1 only represents hydrogen, A and Q1 together very particularly preferably represent C3-C4-alkanediyl, Q1 very particularly preferably represents hydrogen, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, cyclopentyl, cyclohexyl or 4-chlorophenyl, Q2, Q3, Q4 independently of one another very particularly preferably represent hydrogen, methyl or ethyl, Q1 and Q2 together with the carbon atom to which they are attached very particularly preferably represent optionally methyl-, ethyl-, methoxy-, ethoxy-, n-propoxy- or n-butoxy-substituted saturated C5-C6-cycloalkyl in which optionally one ring member is replaced by oxygen, with the proviso that in this case A only represents hydrogen, G very particularly preferably represents hydrogen (a) or represents one of the groups (g), in particular (a), (b) or (c), in which E represents a metal ion or an ammonium ion, L represents oxygen or sulphur and M represents oxygen or sulphur, R1 very particularly preferably represents in each case optionally fluorine or chlorine-substituted C1-C8-alkyl, C2-C8-alkenyl, C1-C2-alkoxy-C1-C2-alkyl, C1-C2-alkylthio-C1-C2-alkyl or cyclopropyl or cyclohexyl, each of which is optionally monosubstituted by fluorine, chlorine, methyl, ethyl or methoxy, very particularly preferably represents phenyl which is optionally monosubstituted by fluorine, chlorine, bromine, cyano, nitro, methyl, ethyl, isopropyl, tert-butyl, methoxy, trifluoromethyl or trifluoromethoxy, very particularly preferably represents thienyl or pyridyl, each of which is optionally monosubstituted by fluorine, chlorine, bromine or methyl, R2 very particularly preferably represents C1-C8-alkyl, C2-C8-alkenyl or C1-C4-alkoxy-C2-alkyl, very particularly preferably represents C3-C6-cycloalkyl which is optionally monosubstituted by methyl, ethyl or methoxy, very particularly preferably represents phenyl or benzyl, each of which is optionally monosubstituted by fluorine, chlorine, cyano, nitro, methyl, ethyl, isopropyl, tert-butyl, methoxy, trifluoromethyl or trifluoromethoxy, R3 very particularly preferably represents methyl or ethyl, each of which is optionally trisubstituted by fluorine or phenyl which is optionally monosubstituted by fluorine, chlorine, bromine, methyl, tert-butyl, methoxy, trifluoromethyl, trifluoromethoxy, cyano or nitro, R4 very particularly preferably represents C1-C4-alkyl, C1-C4-alkoxy, C1-C4-alkylamino, di-(C1-C4-alkyl)amino, C1-C4-alkylthio or represents phenyl, phenoxy or phenylthio, each of which is optionally monosubstituted by fluorine, chlorine, bromine, nitro, cyano, C1-C2-alkoxy, trifluoromethoxy or C1-C3-alkyl, R5 very particularly preferably represents methyl, ethyl, methoxy, ethoxy, methylthio or ethylthio, R6 very particularly preferably represents hydrogen, represents C1-C4-alkyl, C3-C6-cycloalkyl, C1-C4-alkoxy, C3-C4-alkenyl or C1-C4-alkoxy-C1-C4-alkyl, R7 very particularly preferably represents hydrogen, C1-C4-alkyl or C3-C4-alkenyl, R6 and R7 together with the nitrogen atom to which they are attached very particularly preferably represent a C5-alkylene radical in which optionally one methylene group is replaced by oxygen.\",\n \"Het most preferably represents m most preferably represents the number 0 or 1, X most preferably represents hydrogen, methyl or ethyl, Y most preferably represents the group V1 most preferably represents hydrogen, chlorine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, trifluoromethyl or represents 4-chloro-phenoxy, A most preferably represents hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, phenyl or cyclohexyl, B most preferably represents hydrogen, methyl or ethyl, A, B and the carbon atom to which they are attached most preferably represent saturated C5-C6-cycloalkyl in which optionally one ring member is replaced by oxygen and which is optionally monosubstituted by methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, methoxy, ethoxy, n-propoxy, n-butoxy or isobutoxy, with the proviso that in this case Q1 only represents hydrogen, A and Q1 together most preferably represent C3-C4-alkanediyl, Q1 most preferably represents hydrogen, methyl, ethyl, n-propyl, isopropyl or 4-chlorophenyl, Q2 most preferably represents hydrogen, methyl or ethyl, Q3 most preferably represents hydrogen, Q4 most preferably represents hydrogen, Q1 and Q2 together with the carbon atom to which they are attached most preferably represent saturated C5-C6-cycloalkyl, with the proviso that in this case A only represents hydrogen, G most preferably represents hydrogen (a) or represents one of the groups in which L represents oxygen and M represents oxygen or sulphur, R1 most preferably represents C1-C8-alkyl, C1-C2-alkoxy-C1-C2-alkyl or cyclopropyl which is optionally monosubstituted by chlorine, represents phenyl which is optionally monosubstituted by chlorine, represents pyridyl which is optionally monosubstituted by chlorine, R2 most preferably represents C1-C8-alkyl, represents phenyl or benzyl, R3 most preferably represents methyl or ethyl, R6 and R7 together with the nitrogen atom to which they are attached most preferably represent a C5-alkylene radical in which optionally one methylene group is replaced by oxygen.\",\n \"The general or preferred radical definitions or illustrations given above can be combined with one another as desired, i.e.\",\n \"including combinations between the respective ranges and preferred ranges.\",\n \"They apply to the end products and, correspondingly, to precursors and intermediates.\",\n \"Preference according to the invention is given to the compounds of the formula (I) which contain a combination of the meanings given above as being preferred (preferable).\",\n \"Particular preference according to the invention is given to the compounds of the formula (I) which contain a combination of the meanings given above as being particularly preferred.\",\n \"Very particular preference according to the invention is given to the compounds of the formula (I) which contain a combination of the meanings given above as being very particularly preferred.\",\n \"Most preference according to the invention is given to the compounds of the formula (I) which contain a combination of the meanings given above as being most preferred.\",\n \"Particular preference is given to compounds of the formula (I) in which G represents hydrogen.\",\n \"Saturated or unsaturated hydrocarbon radicals, such as alkyl or alkenyl, can in each case be straight-chain or branched as far as this is possible, including in combination with heteroatoms, such as, for example, in alkoxy.\",\n \"Unless indicated otherwise, optionally substituted radicals can be mono- or poly-substituted, where in the case of polysubstitution the substituents can be identical or different.\",\n \"Using, for example, according to process (A) 1-[4-(5-methyl)-2-(4-chloro-phenyl)-thiazolyl]-3-(1-ethoxycarbonyl-cyclohexyl)-propanone as starting material, the course of the process according to the invention can be represented by the reaction scheme below: Using, for example, according to process (B) 1-[4-(5-methyl)-2-(4-chloro-phenyl)-thiazolyl]-4-(l-ethoxycarbonyl-cyclohexyl)-butan-2-one as starting material, the course of the process according to the invention can be represented by the following reaction scheme: Using, for example, according to process (Dα) 2-[4-(5-methyl-2-(3-chloro-phenyl)-thiazolyl]-5,5-dimethylcyclohexane-1,3-dione and pivaloyl chloride as starting materials, the course of the process according to the invention can be represented by the following reaction scheme: Using, for example, according to process (Dβ) 2-[4-(5-ethyl-2-(4-methoxy-phenyl))-thiazolyl]-5,5-dimethyl-cyclohexane-1,3-dione and acetic anhydride as starting materials, the course of the process according to the invention can be represented by the reaction scheme below: Using, for example, according to process (E) 2-[4-(5-methyl-2-phenyl)-thiazolyl]-5,5-dimethyl-cyclohexane-1,3-dione and ethoxyethyl chloroformate as starting materials, the course of the process according to the invention can be represented by the reaction scheme below: Using, for example, according to process (F), 2-[4-(5-methyl-2-(4-fluorophenyl))-thiazolyl]-4,4-dimethyl-cyclopentane-1,3-dione and methyl chloromonothioformate as starting materials, the course of the reaction can be represented as shown below: Using, for example, according to process (G) 2-[4-(5-methyl-3-(4-methyl-phenyl)-thiazolyl]-5,5-dimethyl-cyclohexane-1,3-dione and methanesulphonyl chloride as starting materials, the course of the reaction can be represented by the reaction scheme below: Using, for example, according to process (H) 2-[4-(5-methyl-2-phenyl)-thiazolyl]-4,4-dimethyl-cyclopentane-1,3-dione and 2,2,2-trifluoroethyl methanethio-phosphonyl chloride as starting materials, the course of the reaction can be represented by the reaction scheme below: Using, for example, according to process (I) 2-[4-(5-methyl-2-(4-trifluornethyl-phenyl))-thiazolyl]-5,5-dimethyl-cyclohexane-1,3-dione and NaOH as components, the course of the process according to the invention can be represented by the reaction scheme below: Using, for example, according to process (Jα) 2-[4-(5-methyl-2-(3-trifluoromethyl-phenyl))-thiazolyl]-4,4-dimethyl-cyclohexane-1,3-dione and ethyl isocyanate as starting materials, the course of the reaction can be represented by the reaction scheme below: Using, for example, according to process (Jβ) 2-[4-(5-methyl-2-phenyl)-thiazolyl]-5,5-dimethyl-cyclohexyl-1,3-dione and dimethylcarbamoyl chloride as starting materials, the course of the reaction can be represented by the scheme below: The ketocarboxylic esters of the formula (II) in which A, B, Het, Q1, Q2 and R8 are as defined above, which are required as starting materials in the above process (A), are novel.\",\n \"They can be prepared by methods known in principle.\",\n \"The 5-hetaryl-4-ketocarboxylic esters of the formula (II) are obtained, for example, when 5-hetaryl-4-ketocarboxylic acids of the formula (XIV) in which A, B, Het, Q1 and Q2 are as defined above, are esterified (cf., for example, Organikum, 15th edition, Berlin, 1977,page 499) or alkylated (see Preparation Example).\",\n \"The 5-hetaryl4-ketocarboxylic acids of the formula (XIV) in which A, B, Het, Q1 and Q2 are as defined above, are novel; they can however be prepared by methods known in principle (see Preparation Example).\",\n \"The 5-hetaryl4-ketocarboxylic acids of the formula (XIV) are obtained, for example, when 2-hetaryl-3-oxo-adipic esters of the formula (XV) in which A, B, Het, Q1 and Q2 are as defined above and R8 and R8′ represent alkyl (in particular C1-C8-alkyl) are decarboxylated, if appropriate in the presence of a diluent and if appropriate in the presence of a base or an acid (cf., for example Organikum, 15th edition, Berlin, 1977,pages 519-521).\",\n \"The compounds of the formula (XV) in which A, B, Het, Q1, Q2, R8 and R8′ are as defined above, or, if the compound of the formula (XVII) was used as starting material for the preparation, in which case R8 represents hydrogen, are novel.\",\n \"The compounds of the formula (XV) are obtained, for example, when dicarboxylic monoester chlorides of the formula (XVI), in which A, B, Q1, Q2 and R8 are as defined above and Hal represents chlorine or bromine, or carboxylic anhydrides of the formula (XVII) in which A, B, Q1 and Q2 are as defined above, are acylated with a hetarylacetic ester of the formula (XVIII) in which Het and R8′ are as defined above, in the presence of a diluent and in the presence of a base (cf., for example, M. S. Chambers, E. J. Thomas, D. J. Williams, J. Chem.\",\n \"Soc.\",\n \"Chem.\",\n \"Commun., (1987), 1228,cf.\",\n \"also the Preparation Examples).\",\n \"Some of the compounds of the formulae (XVI) and (XVII) are known compounds of organic chemistry, and/or they can be prepared in a simple manner by methods known in principle.\",\n \"The ketocarboxylic esters of the formula (III) in which A, B, Het, Ql, Q2, Q3, Q4 and R8 are as defined above, which are required as starting materials for the above process (B), are novel.\",\n \"They can be prepared by methods known in principle.\",\n \"The 6-hetaryl-5-ketocarboxylic esters of the formula (III) are obtained, for example, when 6-hetaryl-5-ketocarboxylic acids of the formula (XIX) in which A, B, Het, Q1, Q2, Q3 and Q4 are as defined above, are esterified (cf., for example, Organikum, 15th edition, Berlin, 1977,page 499) or alkylated (see Preparation Example).\",\n \"The 6-aryl-5-ketocarboxylic acids of the formula (XIX) in which A, B, Het, Q1, Q2, Q3 and Q4 are as defined above are novel.\",\n \"They can be prepared by methods known in principle, for example by hydrolysing and decarboxylating substituted 2-hetaryl-3-oxo-heptanedioic esters of the formula (XX) in which A, B, Het, Q1, Q2, Q3 and Q4 are as defined above and R8 and R8′ represent alkyl (preferably C1-C6-alkyl), if appropriate in the presence of a diluent and if appropriate in the presence of a base or an acid (cf., for example, Organikum, 15th edition, Berlin, 1977,pages 519-521).\",\n \"The compounds of the formula (XX) in which A, B, Het, Q1, Q2, Q3, Q4, R8 and R8′ are as defined above, or, if the compound of the formula (XXII) was used as starting material for the preparation, in which case R8 represents hydrogen, are novel.\",\n \"The compounds of the formula (XX) are obtained, for example, when dicarboxylic esters of the formula (XXI), or dicarboxylic anhydrides of the formula (XXII) in which A, B, Q1, Q2, Q3, Q4 and R8 are as defined above, are condensed with a substituted hetarly acetic ester of the formula (XVII) in which Het and R8′ are as defined above, in the presence of a diluent and in the presence of a base.\",\n \"Some of the compounds of the formulae (XXI) and (XXII) are commercially available, some are known, and/or they can be prepared by known processes.\",\n \"Some of the compounds of the formula (XVIII) are novel; however, they can be prepared by processes known in principle: 1.C.\",\n \"S. Rooney et al.\",\n \"J. Med.\",\n \"Chem.\",\n \"1983, 26, 700-714.2.M.\",\n \"S. Malamas et al.\",\n \"J. Med.\",\n \"Chem.\",\n \"1996, 39, 237-245.3.J.\",\n \"L. Collins et al.\",\n \"J. Med.\",\n \"Chem.\",\n \"1998, 41, 5037-5054.4.EP-A-0 177 353.5.EP-A-0 368 592.6.NL-A-6614130.The acyl halides of the formula (VI), carboxylic anhydrides of the formula (V), chloroformic esters or chloroformic thioesters of the formula (VI), chloromonothio-formic esters or chlorodithioformic esters of the formula (V), sulphonyl chlorides of the formula (VII), phosphorus compounds of the formula (IX) and metal hydroxides, metal alkoxides or amines of the formulae (X) and (XI) and isocyanates of the formula (XII) and carbamoyl chlorides or thiocarbamoyl chlorides of the formula (XI) furthermore required as starting materials for carrying out the processes (C), (D), (E), (F), (G), (H), and (I) according to the invention are generally known compounds of organic or inorganic chemistry.\",\n \"Some of the compounds of the formulae (XVI), (XVII) and (XXI) are cornmercially available, some are known, and/or they can be prepared by methods known in principle.\",\n \"The process (A) is characterized in that compounds of the formula (II) in which A, B, Q1, Q2, Het and R8 are as defined above are subjected to an intramolecular condensation in the presence of a base.\",\n \"Suitable for use as diluents in the process (A) according to the invention are all inert organic solvents.\",\n \"Preference is given to using hydrocarbons, such as toluene and xylene, furthermore ethers, such as dibutyl ether, tetrahydrofuran, dioxane, glycol dimethyl ether and diglycol dimethyl ether, moreover polar solvents, such as dimethyl sulphoxide, sulpholane, dimethylformamide, dimethylacetamide and N-methyl-pyrrolidone, and also alcohols, such as methanol, ethanol, propanol, iso-propanol, butanol, iso-butanol and tert-butanol.\",\n \"Suitable bases (deprotonating agents) for carrying out the process (A) according to the invention are all customary proton acceptors.\",\n \"Preference is given to using alkali metal and alkaline earth metal oxides, hydroxides and carbonates, such as sodium hydroxide, potassium hydroxide, magnesium oxide, calcium oxide, sodium carbonate, potassium carbonate and calcium carbonate, which can also be used in the presence of phase-transfer catalysts, such as, for example, triethylbenzylammonium chloride, tetrabutylamrnmonium bromide, Adogen 464 (=methyltrialkyl(C8-C10)ammonium chloride) or TDA 1 (=tris-(methoxyethoxyethyl)-amine).\",\n \"It is furthermore possible to use alkali metals such as sodium or potassium.\",\n \"Furthermore, it is possible to use alkali metal and alkaline earth metal amides and hydrides, such as sodium amide, sodium hydride and calcium hydride, and, moreover, also alkali metal alkoxides, such as sodium methoxide, sodium ethoxide and potassium tert-butoxide.\",\n \"When carrying out the process (A) according to the invention, the reaction temperatures can be varied within a relatively wide range.\",\n \"In general, the process is carried out at temperatures between −80° C. and 180° C., preferably between −50° C. and 120° C. The process (A) according to the invention is generally carried out under atmospheric pressure.\",\n \"When carrying out the process (A) according to the invention, the reaction components of the formula (II) and the deprotonating bases are generally employed in approximately double-equimolar amounts.\",\n \"However, it is also possible to use a relatively large excess (up to 3 mol) of one or the other component.\",\n \"The process (B) is characterized in that compounds of the formula (III) in which A, B, Q1, Q2, Q3, Q4, Het and R8 are as defined above are subjected to an intramolecular condensation in the presence of a diluent and in the presence of a base.\",\n \"Suitable for use as diluents in the process (B) according to the invention are all inert organic solvents.\",\n \"Preference is given to using hydrocarbons, such as toluene and xylene, furthermore ethers, such as dibutyl ether, tetrahydrofuran, dioxane, glycol dimethyl ether and diglycol dimethyl ether, moreover polar solvents, such as dimethyl sulphoxide, sulpholane, dimethylformamide and N-methyl-pyrrolidone.\",\n \"It is furthermore possible to use alcohols, such as methanol, ethanol, propanol, iso-propanol, butanol, iso-butanol and tert-butanol.\",\n \"Suitable bases (deprotonating agents) for carrying out the process (B) according to the invention are all customary proton acceptors.\",\n \"Preference is given to using alkali metal and alkaline earth metal oxides, hydroxides and carbonates, such as sodium hydroxide, potassium hydroxide, magnesium oxide, calcium oxide, sodium carbonate, potassium carbonate and calcium carbonate, which can also be used in the presence of phase-transfer catalysts, such as, for example, triethylbenzylammonium chloride, tetrabutylammonium bromide, Adogen 464 (=methyltrialkyl(C8-C10)ammonium chloride) or TDA 1 (=tris-(methoxyethoxyethyl)-amine).\",\n \"It is furthermore possible to use alkali metals such as sodium or potassium.\",\n \"Furthermore, it is possible to use alkali metal and alkaline earth metal amides and hydrides, such as sodium amide, sodium hydride and calcium hydride, and, moreover, also alkali metal alkoxides, such as sodium methoxide, sodium ethoxide and potassium tert-butoxide.\",\n \"When carrying out the process (B) according to the invention, the reaction temperatures can be varied within a relatively wide range.\",\n \"In general, the process is carried out at temperatures between −80° C. and 180° C., preferably between −50° C. and 120° C. The process (B) according to the invention is generally carried out under atmospheric pressure.\",\n \"When carrying out the process (B) according to the invention, the reaction components of the formula (III) and the deprotonating bases are generally employed in approximately equimolar amounts.\",\n \"However, it is also possible to use a relatively large excess (up to 3 mol) of one or the other component.\",\n \"The process (C-α) is characterized in that compounds of the formulae (I-1-a) to (I-2-a) are in each case reacted with carbonyl halides of the formula (IV), if appropriate in the presence of a diluent and if appropriate in the presence of an acid binder.\",\n \"Suitable diluents for the process (C-60 ) according to the invention are all solvents which are inert to the acyl halides.\",\n \"Preference is given to using hydrocarbons, such as benzine, benzene, toluene, xylene and tetraline, furthermore halogenated hydrocarbons, such as methylene chloride, chloroform, carbon tetrachloride, chlorobenzene and o-dichlorobenzene, moreover ketones, such as acetone and methyl isopropyl ketone, furthermore ethers, such as diethyl ether, tetrahydrofuran and dioxane, additionally carboxylic esters, such as ethyl acetate, nitriles, such as acetonitrile, and also strongly polar solvents, such as dimethylformamide, dimethylacetamide, dimethyl sulphoxide and sulpholane.\",\n \"The hydrolytic stability of the acyl halide permitting, the reaction can also be carried out in the presence of water.\",\n \"Suitable acid binders for the reaction according to the process (C-α) according to the invention are all customary acid acceptors.\",\n \"Preference is given to using tertiary amines, such as triethylamine, pyridine, diazabicyclooctane (DABCO), diazabi-cycloundecene (DBU), diazabicyclononene (DBN), Hünig base or N,N-dimethylaniline, furthermore alkaline earth metal oxides, such as magnesium oxide and calcium oxide, moreover alkali metal and alkaline earth metal carbonates, such as sodium carbonate, potassium carbonate and calcium carbonate, and also alkali metal hydroxides, such as sodium hydroxide and potassium hydroxide.\",\n \"The reaction temperatures in the process (C-α) according to the invention can be varied within a relatively wide range.\",\n \"In general, the process is carried out at temperatures between −20° C. and +150° C., preferably between 0° C. and 100° C. When carrying out the process (C-α) according to the invention, the starting materials of the formulae (I-1-a) to (I-2-a) and the carbonyl halide of the formula (IV) are generally each employed in approximately equivalent amounts.\",\n \"However, it is also possible to use a relatively large excess (up to 5 mol) of the carbonyl halide.\",\n \"Work-up is carried out by customary methods.\",\n \"The process (C-β) is characterized in that compounds of the formulae (I1-a) to (I2-a) are reacted with carboxylic anhydrides of the formula (V), if appropriate in the presence of a diluent and if appropriate in the presence of an acid binder.\",\n \"Suitable diluents for use in the process (C-β) according to the invention are preferably those solvents which are also preferred when acyl halides are used.\",\n \"Furthermore, it is also possible for a carboxylic anhydride employed in excess to act simultaneously as diluent.\",\n \"Suitable acid binders, which are, if appropriate, added in the process (C-β), are preferably those acid binders which are also preferred when acyl halides are used.\",\n \"The reaction temperatures in the process (C-β) according to the invention can be varied within a relatively wide range.\",\n \"In general, the process is carried out at temperatures between −20° C. and +150° C., preferably between 0° C. and 100° C. When carrying out the process (C-β) according to the invention, the starting materials of the formulae (I-1-a) to (I2-a) and the carboxylic anhydride of the formula (V) are generally each employed in approximately equivalent amounts.\",\n \"However, it is also possible to use a relatively large excess (up to 5 mol) of the carboxylic anhydride.\",\n \"Work-up is carried out by customary methods.\",\n \"In general, diluent and excess carboxylic anhydride and the carboxylic acid that is formed are removed by distillation or by washing with an organic solvent or with water.\",\n \"The process (D) is characterized in that compounds of the formulae (I1-a) to (I2-a) are in each case reacted with chlorofornic esters or chloroformic thiolesters of the formula (VI), if appropriate in the presence of a diluent and if appropriate in the presence of an acid binder.\",\n \"Suitable acid binders for the reaction according to process (D) according to the invention are all customary acid acceptors.\",\n \"Preference is given to using tertiary amines, such as triethylamine, pyridine, DABCO, DBU, DBA, Hünig base and N,N-dimethyl-aniline, furthermore alkaline earth metal oxides, such as magnesium oxide and calcium oxide, moreover alkali metal and alkaline earth metal carbonates, such as sodium carbonate, potassium carbonate and calcium carbonate, and also alkali metal hydroxides, such as sodium hydroxide and potassium hydroxide.\",\n \"Suitable diluents for the process (D) according to the invention are all solvents which are inert to the chloroformic esters or chloroformic thiolesters used.\",\n \"Preference is given to using hydrocarbons, such as benzine, benzene, toluene, xylene and tetraline, furthermore halogenated hydrocarbons, such as methylene chloride, chloroform, carbon tetrachloride, chlorobenzene and o-dichlorobenzene, moreover ketones, such as acetone and methyl isopropyl ketone, furthermore ethers, such as diethyl ether, tetrahydrofuran and dioxane, additionally carboxylic esters, such as ethyl acetate, nitriles, such as acetonitrile, and also strongly polar solvents, such as dimethylformamide, dimethylacetamide, dimethyl sulphoxide and sulpholane.\",\n \"When carrying out the process (D) according to the invention, the reaction temperatures can be varied within a relatively wide range.\",\n \"If the process is carried out in the presence of a diluent and an acid binder, the reaction temperatures are generally between −20° C. and +100° C., preferably between 0° C. and 50° C. The process (D) according to the invention is generally carried out under atmospheric pressure.\",\n \"When carrying out the process (D) according to the invention, the starting materials of the formulae (I-1-a) to (I2-a) and the appropriate chloroformic ester or chloroformic thiolester of the formula (VI) are generally each employed in approximately equivalent amounts.\",\n \"However, it is also possible to use a relatively large excess (up to 2 mol) of one or the other component.\",\n \"Work-up is carried out by customary methods.\",\n \"In general, precipitated salts are removed and the reaction mixture that remains is concentrated by stripping off the diluent.\",\n \"Process (E) according to the invention is characterized in that compounds of the formulae (I1-a) to (I-2-a) are in each case reacted with compounds of the formula (VII), in the presence of a diluent and, if appropriate, in the presence of an acid binder.\",\n \"In the preparation process (E), about 1 mol of chloromonothioformic ester or chlorodithioformic ester of the formula (VII) is employed per mole of starting material of the formulae (I-1-a) to (I2-a), at from 0 to 120° C., preferably at from 20 to 60° C. Suitable diluents, which are added, if appropriate, are all inert polar organic solvents, such as nitrites, esters, ethers, amides, sulphones, sulphoxides, and also halogenoalkanes.\",\n \"Preference is given to using acetonitrile, ethyl acetate, dimethyl sulphoxide, tetra-hydrofuran, dimethylformamide or methylene chloride.\",\n \"If, in a preferred embodiment, the enolate salt of the compounds (I1-a) to (I-2-a) is prepared by adding strong deprotonating agents, such as, for example, sodium hydride or potassium tert-butoxide, the further addition of acid binders can be dispensed with.\",\n \"If acid binders are used, these are customary inorganic or organic bases, for example sodium hydroxide, sodium carbonate, potassium carbonate, pyridine, triethylamine.\",\n \"The reaction can be carried out at atmospheric pressure or under elevated pressure and is preferably carried out at atmospheric pressure.\",\n \"Work-up is carried out by customary methods.\",\n \"The process (F) according to the invention is characterized in that compounds of the formulae (I1-a) to (I2-a) are in each case reacted with sulphonyl chlorides of the formula (VII), if appropriate in the presence of a diluent and if appropriate in the presence of an acid binder.\",\n \"In the preparation process (F), about 1 mol of sulphonyl chloride of the formula.\",\n \"(VII) is employed per mole of starting material of the formulae (I-1-a) to (I2-a), at from −20 to 150° C., preferably at from 20 to 70° C. Suitable diluents which are added, if appropriate, are all inert polar organic solvents, such as nitrites, esters, ethers, amides, sulphones, sulphoxides, or halogenated hydrocarbons, such as methylene chloride.\",\n \"Preference is given to using acetonitrile, ethyl acetate, dimethyl sulphoxide, tetra-hydrofuran, dimethylformamide, methylene chloride.\",\n \"If, in a preferred embodiment, the enolate salt of the compounds (I1-a) to (I-2-a) is prepared by adding strong deprotonating agents (such as, for example, sodium hydride or potassium tert-butoxide), the further addition of acid binders can be dispensed with.\",\n \"If acid binders are used, these are customary inorganic or organic bases, for example sodium hydroxide, sodium carbonate, potassium carbonate, pyridine, triethylamine.\",\n \"The reaction can be carried out at atmospheric pressure or under elevated pressure and is preferably carried out at atmospheric pressure.\",\n \"Work-up is carried out by customary methods.\",\n \"The process (G) according to the invention is characterized in that compounds of the formulae (I1-a) to (I2-a) are in each case reacted with phosphorus compounds of the formula (IX), if appropriate in the presence of a diluent and if appropriate in the presence of an acid binder.\",\n \"In the preparation process (G), to obtain compounds of the formulae (1-1-e) to (1-2-e), 1 to 2,preferably 1 to 1.3,mol of the phosphorus compound of the formula (IX) are employed per mole of the compounds (I1-a) to (I-2-a), at temperatures between −40° C. and 150° C., preferably between −10 and 110° C. Suitable diluents which are added, if appropriate, are all inert polar organic solvents, such as ethers, amides, nitrites, alcohols, sulphides, sulphones, sulphoxides, etc.\",\n \"Preference is given to using acetonitrile, dimethyl sulphoxide, tetrahydrofuran, dimethylformamide, methylene chloride.\",\n \"Suitable acid binders which are added, if appropriate, are customary inorganic or organic bases, such as hydroxides, carbonates or amines.\",\n \"Examples which may be mentioned are sodium hydroxide, sodium carbonate, potassium carbonate, pyridine, triethylamine.\",\n \"The reaction can be carried out at atmospheric pressure or under elevated pressure and is preferably carried out at atmospheric pressure.\",\n \"Work-up is carried out by customary methods of organic chemistry.\",\n \"The end products obtained are preferably purified by crystallization, chromatographic purification or by “incipient distillation”, i.e.\",\n \"removal of the volatile components under reduced pressure.\",\n \"The process (H) is characterized in that compounds of the formulae (I-1-a) to (I2-a) are reacted with metal hydroxides or metal alkoxides of the formula (X) or amines of the formula (XI), if appropriate in the presence of a diluent.\",\n \"Suitable for use as diluents in the process (H) according to the invention are, preferably, ethers, such as tetrahydrofuran, dioxane, diethyl ether, or else alcohols, such as methanol, ethanol, isopropanol, and also water.\",\n \"The process (H) according to the invention is generally carried out under atmospheric pressure.\",\n \"The reaction temperatures are generally between −20° C. and 100° C., preferably between 0° C. and 50° C. The process (I) according to the invention is characterized in that compounds of the formulae (I-1-a) to (I2-a) are in each case reacted with compounds of the formula (XII) (I-α), if appropriate in the presence of a diluent and if appropriate in the presence of a catalyst, or (I-β) with compounds of the formula (XII), if appropriate in the presence of a diluent and if appropriate in the presence of an acid binder.\",\n \"In the preparation process (I-α), about 1 mol of isocyanate of the formula (XII) is employed per mole of starting material of the formulae (I1-a) to (I2-a), at from 0 to 100° C., preferably at from 20 to 50° C. Suitable diluents which are added, if appropriate, are all inert organic solvents, such as ethers, amides, nitriles, sulphones, sulphoxides.\",\n \"If appropriate, catalysts may be added to accelerate the reaction.\",\n \"Very advantageously, the catalysts which are employed are organotin compounds, such as, for example, dibutyltin dilaurate.\",\n \"The reaction is preferably carried out at atmospheric pressure.\",\n \"In preparation process (I-β), about 1 mol of carbamoyl chloride of the formula (XIII) is reacted per mole of starting material of the formulae (I-1-a) to (I-2-a) at from −20 to 150° C., preferably at from 0 to 70° C. Suitable diluents which are added, if appropriate, are all inert polar organic solvents, such as nitrites, esters, ethers, amides, sulphones, sulphoxides or halogenated hydrocarbons.\",\n \"Preference is given to using acetonitrile, ethyl acetate, dimethyl sulphoxide, tetrahydrofuran, dimethylformamide or methylene chloride.\",\n \"If, in a preferred embodiment, the enolate salt of the compounds (I-1-a) to (I2-a) is prepared by adding strong deprotonating agents (such as, for example, sodium hydride or potassium tert-butoxide), the addition of further acid binders can be dispensed with.\",\n \"The active compounds are suitable for controlling animal pests, in particular insects, arachnids and nematodes, which are encountered in agriculture, in forestry, in the protection of stored products and of materials, and in the hygiene sector, and have good plant tolerance and favourable toxicity to warm-blooded animals.\",\n \"They may be preferably employed as plant protection agents.\",\n \"They are active against normally sensitive and resistant species and against all or some stages of development.\",\n \"The abovementioned pests include: From the order of the Isopoda, for example, Oniscus asellus, Armadillidium vulgare and Porcellio scaber.\",\n \"From the order of the Diplopoda, for example, Blaniulus guttulatus.\",\n \"From the order of the Chilopoda, for example, Geophilus carpophagus and Scutigera spp.\",\n \"From the order of the Symphyla, for example, Scutigerella immaculata.\",\n \"From the order of the Thysanura, for example, Lepisma saccharina.\",\n \"From the order of the Collembola, for example, Onychiurus armatus.\",\n \"From the order of the Orthoptera, for example, Acheta domesticus, Gryllotalpa spp., Locusta migratoria migratorioides, Melanoplus spp.\",\n \"and Schistocerca gregaria.\",\n \"From the order of the Blattaria, for example, Blatta orientalis, Periplaneta americana, Leucophaea maderae, Blattella germanica.\",\n \"From the order of the Dermaptera, for example, Forficula auricularia.\",\n \"From the order of the Isoptera, for example, Reticulitermes spp.\",\n \"From the order of the Phthiraptera, for example, Pediculus humanus corporis, Haematopinus spp., Linognathus spp., Trichodectes spp.\",\n \"and Damalinia spp.\",\n \"From the order of the Thysanoptera, for example, Hercinothrips femoralis, Thrips tabaci, Thrips palmi and Frankliniella accidentalis.\",\n \"From the order of the Heteroptera, for example, Eurygaster spp., Dysdercus intermedius, Piesma quadrata, Cimex lectularius, Rhodnius prolixus and Triatoma spp.\",\n \"From the order of the Homoptera, for example, Aleurodes brassicae, Bemisia tabaci, Trialeurodes vaporariorum, Aphis gossypii, Brevicoryne brassicae, Cryptomyzus ribis, Aphis fabae, Aphis pomi, Eriosoma lanigerum, Hyalopterus arundinis, Phylloxera vastatrix, Pemphigus spp., Macrosiphum avenae, Myzus spp., Phorodon humuli, Rhopalosiphum padi, Empoasca spp., Euscelis bilobatus, Nephotettix cincticeps, Lecanium corni, Saissetia oleae, Laodelphax striatellus, Nilaparvata lugens, Aonidiella aurantii, Aspidiotus hederae, Pseudococcus spp.\",\n \"and Psylla spp.\",\n \"From the order of the Lepidoptera, for example, Pectinophora gossypiella, Bupalus piniarius, Cheimatobia brumata, Lithocolletis blancardella, Hyponomeuta padella, Plutella xylostella, Malacosoma neustria, Euproctis chrysorrhoea, Lymantria spp., Bucculatrix thurberiella, Phyllocnistis citrella, Agrotis spp., Euxoa spp., Feltia spp., Earias insulana, Heliothis spp., Mamestra brassicae, Panolis flammea, Spodoptera spp., Trichoplusia ni, Carpocapsa pomonella, Pieris spp., Chilo spp., Pyrausta nubilalis, Ephestia kuehniella, Galleria mellonella, Tineola bisselliella, Tinea pellionella, Hofmannophila pseudospretella, Cacoecia podana, Capua reticulana, Choristoneura fumiferana, Clysia ambiguella, Homona magnanima, Tortrix viridana, Cnaphalocerus spp., Oulema oryzae.\",\n \"From the order of the Coleoptera, for example, Anobium punctatum, Rhizopertha dominica, Bruchidius obtectus, Acanthoscelides obtectus, Hylotrupes bajulus, Agelastica alni, Leptinotarsa decemlineata, Phaedon cochleariae, Diabrotica spp., Psylliodes chrysocephala, Epilachna varivestis, Atomaria spp., Oryzaephilus surinamensis, Anthonomus spp., Sitophilus spp., Otiorrhynchus sulcatus, Cosmopolites sordidus, Ceuthorrhynchus assimilis, Hypera postica, Dennestes spp., Trogoderma spp., Anthrenus spp., Attagenus spp., Lyctus spp., Meligethes aeneus, Ptinus spp., Niptus hololeucus, Gibbium psylloides, Tribolium spp., Tenebrio molitor, Agriotes spp., Conoderus spp., Melolontha melolontha, Amphimallon solstitialis, Costelytra zealandica and Lissorhoptrus oryzophilus.\",\n \"From the order of the Hymenoptera, for example, Diprion spp., Hoplocampa spp., Lasius spp., Monomorium pharaonis and Vespa spp.\",\n \"From the order of the Diptera, for example, Aedes spp., Anopheles spp., Culex spp., Drosophila melanogaster, Musca spp., Fannia spp., Calliphora erythrocephala, Lucilia spp., Chrysomyia spp., Cuterebra spp., Gastrophilus spp., Hyppobosca spp., Stomoxys spp., Oestrus spp., Hypoderma spp., Tabanus spp., Tannia spp., Bibio hortulanus, Oscinella frit, Phorbia spp., Pegomyia hyoscyami, Ceratitis capitata, Dacus oleae, Tipula paludosa, Hylemyia spp.\",\n \"and Liriomyza spp.\",\n \"From the order of the Siphonaptera, for example, Xenopsylla cheopis and Ceratophyllus spp.\",\n \"From the class of the Arachnida, for example, Scorpio maurus, Latrodectus mactans, Acarus siro, Argas spp., Omithodoros spp., Dermanyssus gallinae, Eriophyes ribis, Phyllocoptruta oleivora, Boophilus spp., Rhipicephalus spp., Amblyomma spp., Hyalomma spp., Ixodes spp., Psoroptes spp., Chorioptes spp., Sarcoptes spp., Tarsonemus spp., Bryobia praetiosa, Panonychus spp., Tetranychus spp., Hemitarsonemus spp., Brevipalpus spp.\",\n \"The phytoparasitic nematodes include, for example, Pratylenchus spp., Radopholus similis, Ditylenchus dipsaci, Tylenchulus semipenetrans, Heterodera spp., Globodera spp., Meloidogyne spp., Aphelenchoides spp., Longidorus spp., Xiphinema spp., Trichodorus spp., Bursaphelenchus spp.\",\n \"If appropriate, the compounds according to the invention can, at certain concentrations or application rates, also be used as herbicides or microbicides, for example as fungicides, antimycotics and bactericides.\",\n \"If appropriate, they can also be employed as intermediates or precursors for the synthesis of other active compounds.\",\n \"All plants and plant parts can be treated in accordance with the invention.\",\n \"Plants are to be understood as meaning in the present context all plants and plant populations such as desired and undesired wild plants or crop plants (including naturally occurring crop plants).\",\n \"Crop plants can be plants which can be obtained by conventional plant breeding and optimization methods or by biotechnological and recombinant methods or by combinations of these methods, including the transgenic plants and inclusive of the plant cultivars protectable or not protectable by plant breeders rights.\",\n \"Plant parts are to be understood as meaning all parts and organs of plants above and below the ground, such as shoot, leaf, flower and root, examples which may be mentioned being leaves, needles, stalks, stems, flowers, fruit bodies, fruits, seeds, roots, tubers and rhizomes.\",\n \"The plant parts also include harvested material, and vegetative and generative propagation material, for example cuttings, tubers, rhizomes, offsets and seeds.\",\n \"Treatment according to the invention of the plants and plant parts with the active compounds is carried out directly or by allowing the compounds to act on the surroundings, environment or storage space by the customary treatment methods, for example by immersion, spraying, evaporation, fogging, scattering, painting on and, in the case of propagation material, in particular in the case of seeds, also by applying one or more coats.\",\n \"The active compounds can be converted to the customary formulations, such as solutions, emulsions, wettable powders, suspensions, powders, dusts, pastes, soluble powders, granules, suspension-emulsion concentrates, natural and synthetic materials impregnated with active compound and microencapsulations in polymeric substances.\",\n \"These formulations are produced in a known manner, for example by mixing the active compounds with extenders, that is, liquid solvents, and/or solid carriers, optionally with the use of surfactants, that is emulsifiers and/or dispersants, and/or foam-formers.\",\n \"If the extender used is water, it is also possible to employ for example organic solvents as auxiliary solvents.\",\n \"Essentially, suitable liquid solvents are: aromatics such as xylene, toluene or alkylnaphthalenes, chlorinated aromatics or chlorinated aliphatic hydrocarbons such as chlorobenzenes, chloroethylenes or methylene chloride, aliphatic hydrocarbons such as cyclohexane or paraffins, for example petroleum fractions, mineral and vegetable oils, alcohols such as butanol or glycol and also their ethers and esters, ketones such as acetone, methyl ethyl ketone, methyl isobutyl ketone or cyclohexanone, strongly polar solvents such as dimethylformamide and dimethyl sulphoxide, and also water.\",\n \"As solid carriers there are suitable: for example ammonium salts and ground natural minerals such as kaolins, clays, talc, chalk, quartz, attapulgite, montmorillonite or diatomaceous earth, and ground synthetic minerals, such as highly disperse silica, alumina and silicates; as solid carriers for granules there are suitable: for example crushed and fractionated natural rocks such as calcite, marble, pumice, sepiolite and dolomite, and also synthetic granules of inorganic and organic meals, and granules of organic material such as sawdust, coconut shells, maize cobs and tobacco stalks; as emulsifiers and/or foam-formers there are suitable: for example nonionic and anionic emulsifiers, such as polyoxyethylene fatty acid esters, polyoxyethylene fatty alcohol ethers, for example alkylaryl polyglycol ethers, alkylsulphonates, alkyl sulphates, arylsulphonates and also protein hydrolysates; as dispersants there are suitable: for example lignin-sulphite waste liquors and methylcellulose.\",\n \"Tackifiers such as carbox),methylcellulose and natural and synthetic polymers in the form of powders, granules or latices, such as gum arabic, polyvinyl alcohol and polyvinyl acetate, as well as natural phospholipids such as cephalins and lecithins, and synthetic phospholipids, can be used in the formulations.\",\n \"Other possible additives are mineral and vegetable oils.\",\n \"It is possible to use colorants such as inorganic pigments, for example iron oxide, titanium oxide and Prussian Blue, and organic dyestuffs, such as alizarin dyestuffs, azo dyestuffs and metal phthalocyanine dyestuffs, and trace nutrients such as salts of iron, manganese, boron, copper, cobalt, molybdenum and zinc.\",\n \"The formulations generally comprise between 0.1 and 95% by weight of active compound, preferably between 0.5 and 90%.\",\n \"The active compounds according to the invention can be employed formulations in their as such or as a mixture with known fungicides, bactericides, acaricides, nematicides or insecticides in order to increase the activity spectrum or avoid the development of resistance.\",\n \"In many cases synergistic effects are achieved, ie.\",\n \"the efficacy of the mixture is greater than the efficacy of the individual components.\",\n \"Favourable examples of co-components in mixtures are the following compounds: Fungicides: aldimorph, ampropylfos, ampropylfos-potassium, andoprim, anilazine, azaconazole, azoxystrobin, benalaxyl, benodanil, benomyl, benzamacril, benzamacryl-isobutyl, bialaphos, binapacryl, biphenyl, bitertanoi, blasticidin-S, bromuconazole, bupirimate, buthiobate, calcium polysulphide, capsimycin, captafol, captan, carbendazim, carboxin, carvon, quinomethionate, chlobenthiazone, chlorfenazole, chloroneb, chloropicrin, chlorothalonil, chlozolinate, clozylacon, cufraneb, cymoxanil, cyproconazole, cyprodinil, cyprofuram, debacarb, dichlorophen, diclobutrazole, diclofluanid, diclomezine, dicloran, diethofencarb, difenoconazole, dimethirimol, dimethomorph, diniconazole, diniconazole-M, dinocap, diphenylamine, dipyrithione, ditalimfos, dithianon, dodemorph, dodine, drazoxolon, edifenphos, epoxiconazole, etaconazole, ethirimol, etridiazole, famoxadon, fenapanil, fenarimol, fenbuconazole, fenfuram, fenitropan, fenpiclonil, fenpropidin, fenpropimorph, fentin acetate, fentin hydroxide, ferbam, ferimzone, fluazinam, flumetover, fluoromide, fluquinconazole, flurprimidol, flusilazole, flusulfamide, flutolanil, flutriafol, folpet, fosetyl-aluminium, fosetyl-sodium, fthalide, fuberidazole, furalaxyl, furametpyr, furcarbonil, furconazole, furconazole-cis, furmecyclox, guazatine, hexachlorobenzene, hexaconazole, hymexazole, imazalil, imibenconazole, iminoctadine, iminoctadine albesilate, iminoctadine triacetate, iodocarb, ipconazole, iprobenfos (IBP), iprodione, irumamycin, isoprothiolane, isovaledione, kasugamycin, kresoxim-methyl, copper preparations, such as: copper hydroxide, copper naphthenate, copper oxychloride, copper sulphate, copper oxide, oxine-copper and Bordeaux mixture, mancopper, mancozeb, maneb, meferimzone, mepanipyrim, mepronil, metalaxyl, metconazole, methasulfocarb, methfuroxam, metiram, metomeclam, metsulfovax, mildiomycin, myclobutanil, myclozolin, nickel dimethyldithiocarbamate, nitrothal-isopropyl, nuarimol, ofurace, oxadixyl, oxamocarb, oxolinic acid, oxycarboxim, oxyfenthiin, paclobutrazole, pefurazoate, penconazole, pencycuron, phosdiphen, picoxystrobin, pimaricin, piperalin, polyoxin, polyoxorim, probenazole, prochloraz, procymidone, propamocarb, propanosine-sodium, propiconazole, propineb, pyraclostrobin, pyrazophos, pyrifenox, pyrimethanil, pyroquilon, pyroxyfur, quinconazole, quintozene (PCNB), sulphur and sulphur preparations, tebuconazole, tecloftalam, tecnazene, tetcyclacis, tetraconazole, thiabendazole, thicyofen, thifluzamide, thiophanate-methyl, thiram, tioxyrmid, tolclofos-methyl, tolyfluanid, triadimefon, triadimenol, triazbutil, triazoxide, trichlamide, tricyclazole, tridemorph, trifloxystrobin, triflurnizole, triforine, triticonazole, uniconazole, validamycin A, vinclozolin, viniconazole, zarilamide, zineb, ziram and also Dagger G, OK-8705, OK-8801, α-(1,1-dimethylethyl)-β-(2-phenoxyethyl)-1H-1,2,4-triazole-1-ethanol, α-(2,4-dichlorophenyl)-β-fluoro-β-propyl-1H-1,2,4-triazole-1-ethanol, α-(2,4-dichlorophenyl)-β-methoxy-α-methyl-1H-1,2,4-triazole-1-ethanol, α-(5-methyl-1,3-dioxan-5-yl)-β-[[4-(trifluoromethyl)-phenyl]-methylene]-1H-1,2,4-triazole-1-ethanol, (5RS,6RS)-6-hydroxy-2,2,7,7-tetramethyl-5-(1H-1,2,4-triazol-1-yl)-3-octanone, (E)-α-(methoxyimino)-N-methyl-2-phenoxy-phenylacetamide, 1-isopropyl {2-methyl-1-[[[1-(4-methylphenyl)-ethyl]-amino]-carbonyl]-propyl}-carbamate, 1-(2,4-dichlorophenyl)-2-(1H-1,2,4-triazol-1-yl)-ethanone-O-(phenylmethyl)-oxime, 1-(2-methyl-1-naphthalenyl)-1H-pyrrol-2,5-dione, 1-(3,5-dichlorophenyl)-3-(2-propenyl)-2,5-pyrrolidindione, 1-[(diiodomethyl)-sulphonyl]4-methyl-benzene, 1-[[2-(2,4-dichlorophenyl)-1,3-dioxolan-2-yl]-methyl]-1H-imidazole, 1-[[2-(4-chlorophenyl)-3-phenyloxiranyl]-methyl]-1H-1,2,4-triazole, 1-[1-[2-[(2,4-dichlorophenyl)-methoxy]-phenyl]-ethenyl]-1H-imidazole, 1-methyl-5-nonyl-2-(phenylmethyl)-3-pyrrolidinole, 2′,6′-dibromo-2-methyl4′-trifluoromethoxy4′-trifluoro-methyl-1,3-thiazole-5-carboxanilide, 2,2-dichloro-N-[1-(4-chlorophenyl)-ethyl]-1-ethyl-3-methyl-cyclopropanecarboxamide, 2,6-dichloro-5-(methylthio)4-pyrimidinyl-thiocyanate, 2,6-dichloro-N-(4-trifluoromethylbenzyl)-benzamide, 2,6-dichloro-N-[[4-(trifluoromethyl)-phenyl]-methyl]-benzamide, 2-(2,3,3-triiodo-2-propenyl)-2H-tetrazole, 2-[(1-methylethyl)-sulphonyl]-5-(trichloromethyl)-1,3,4-thiadiazole, 2-[[6-deoxy4-O-(4-O-methyl-β-D-glycopyranosyl)-α-D-glucopyranosyl]-amino]-4-methoxy-1H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile, 2-aminobutane, 2-bromo-2-(bromomethyl)-pentanedinitrile, 2-chloro-N-(2,3-dihydro-1,1,3-trimethyl-1H-inden4yl)-3-pyridinecarboxamide, 2-chloro-N-(2,6-dimethylphenyl)-N-(isothiocyanatomethyl)-acetamide, 2-phenylphenol (OPP), 3,4-dichloro-1-[4-(difluoromethoxy)-phenyl]-1H-pyrrol-2,5-dione, 3,5-dichloro-N-[cyano[(1-methyl-2-propynyl)-oxy]-methyl]-benzamide, 3-(1,1-dimethylpropyl-1-oxo-1H-indene-2-carbonitrile, 3-[2-(4-chlorophenyl)-5-ethoxy-3-isoxazolidinyl]-pyridine, 4-chloro-2-cyano-N,N-dimethyl-5-(4-methylphenyl)-1H-imidazole-1-sulphonamide, 4-methyl-tetrazolo[1,5-a]quinazolin-5(4H)-one, 8-(1,1-dimethylethyl)-N-ethyl-N-propyl-1,4-dioxaspiro[4.5]decane-2-methanamine, 8-hydroxyquinoline sulphate, 9H-xanthene-2-[(phenylamino)-carbonyl]-9-carboxylic hydrazide, bis-(1-methylethyl)-3-methyl4-[(3-methylbenzoyl)-oxy]-2,5-thiophenedicarboxylate, cis-1-(4-chlorophenyl)-2-(1H-1,2,4-triazol-1-yl)-cycloheptanol, cis4-[3-[4-(1,1-dimethylpropyl)-phenyl-2-methylpropyl]-2,6-dimethyl-morpholine hydrochloride, ethyl [(4-chlorophenyl)-azo]-cyanoacetate, potassium bicarbonate, methanetetrathiol-sodium salt, methyl 1-(2,3-dihydro-2,2-dimethyl-1H-inden-1-yl)-1H-imidazole-5-carboxylate, methyl N-(2,6-dimethylphenyl)-N-(5-isoxazolylcarbonyl)-DL-alaninate, methyl N-(chloroacetyl)-N-(2,6-dimethylphenyl)-DL-alaninate, N-(2,3-dichloro-4-hydroxyphenyl)-1-methyl-cyclohexanecarboxamide, N-(2,6-dimethylphenyl)-2-methoxy-N-(tetrahydro-2-oxo-3-furanyl)-acetamide, N-(2,6-dimethylphenyl)-2-methoxy-N-(tetrahydro-2-oxo-3-thienyl)-acetamide, N-(2-chloro-4-nitrophenyl)-4-methyl-3-nitro-benzenesulphonamide, N-(4-cyclohexylphenyl)-1,4,5,6-tetrahydro-2-pyrimidinamine, N-(4-hexylphenyl)-1,4,5,6-tetrahydro-2-pyrimidinamine, N-(5-chloro-2-methylphenyl)-2-methoxy-N-(2-oxo-3-oxazolidinyl)-acetamide, N-(6-methoxy)-3-pyridinyl-cyclopropanecarboxamide, N-[2,2,2-trichloro-1-[(chloroacetyl)-amino]-ethyl]-benzamide, N-[3-chloro-4,5-bis(2-propinyloxy)-phenyl]-N′-methoxy-methanimidamide, N-formyl-N-hydroxy-DL-alanine-sodium salt, O,O-diethyl [2-(dipropylamino)-2-oxoethyl]-ethylphosphoramidothioate,.\",\n \"O-methyl S-phenyl phenylpropylphosphoramidothioate, S-methyl 1,2,3-benzothiadiazole-7-carbothioate, spiro[2H]-1-benzopyrane-2,1′(3′H)-isobenzofuran]-3′-one, 4-[(3,4-dimethoxyphenyl)-3-(4-fluorophenyl)-acryloyl]-morpholine.\",\n \"Bactericides: bronopol, dichlorophen, nitrapyrin, nickel dimethyldithiocarbamate, kasugamycin, octhilinone, furancarboxylic acid, oxytetracyclin, probenazole, streptomycin, tecloftalam, copper sulphate and other copper preparations.\",\n \"Insecticides/Acaricides/Nematicides: abamectin, acephate, acetamiprid, acrinathrin, alanycarb, aldicarb, aldoxycarb, alpha-cypermethrin, alphamethrin, amitraz, avermectin, AZ 60541, azadirachtin, azamethiphos, azinphos A, azinphos M, azocyclotin, Bacillus popilliae, Bacillus sphaericus, Bacillus subtilis, Bacillus thuringiensis, baculoviruses, Beauveria bassiana, Beauveria tenella, bendiocarb, benfuracarb, bensultap, benzoximate, betacyfluthrin, bifenazate, bifenthrin, bioethanomethrin, biopermethrin, bistrifluron, BPMC, bromophos A, bufencarb, buprofezin, butathiofos, butocarboxim, butylpyridaben, cadusafos, carbaryl, carbofuran, carbophenothion, carbosulfan, cartap, chloethocarb, chlorethoxyfos, chlorfenapyr, chlorfenvinphos,- chlorfluazuron, chlormephos, chlorpyrifos, chlorpyrifos M, chlovaporthrin, chromafenozide, cis-resmethrin, cispertmethrin, clocythrin, cloethocarb, clofentezine, clothianidine, cyanophos, cycloprene, cycloprothrin, cyfluthrin, cyhalothrin, cyhexatin, cypermethrin, cyromazine, deltamethrin, demeton M, demeton S, demeton-S-methyl, diafenthiuron, diazinon, dichlorvos, diflubenzuron, dimethoate, dimethylvinphos, diofenolan, disulfoton, docusat-sodium, dofenapyn, eflusilanate, emamectin, empenthrin, endosulfan, Entomopfthora spp., esfenvalerate, ethiofencarb, ethion, ethoprophos, etofenprox, etoxazole, etrimfos, fenamiphos, fenazaquin, fenbutatin oxide, fenitrothion, fenothiocarb, fenoxacrim, fenoxycarb, fenpropathrin, fenpyrad, fenpyrithrin, fenpyroximate, fenvalerate, fipronil, fluazinam, fluazuron, flubrocythrinate, flucycloxuron, flucythrinate, flufenoxuron, flumethrin, flutenzine, fluvalinate, fonophos, fosmethilan, fosthiazate, fubfenprox, furathiocarb, granulosis viruses, halofenozide, HCH, heptenophos, hexaflumuron, hexythiazox, hydroprene, imidacloprid, indoxacarb, isazofos, isofenphos, isoxathion, ivermectin, nuclear polyhedrosis viruses, lambda-cyhalothrin, lufenuron, malathion, mecarbam, metaldehyde, methamidophos, Metharhizium anisopliae, metharhizium flavoviride, methidathion, methiocarb, methoprene, methomyl, methoxyfenozide, metolcarb, metoxadiazone, mevinphos, milbemectin, milbemycin, monocrotophos, naled, nitenpyram, nithiazine, novaluron, omethoate, oxamyl, oxydemethon M, Paecilomyces fumosoroseus, parathion A, parathion M, permethrin, phenthoate, phorate, phosalone, phosmet, phosphamidon, phoxim, pirimicarb, pirimiphos A, pirimiphos M, profenofos, promecarb, propargite, propoxur, prothiofos, prothoate, pymetrozine, pyraclofos, pyresmethrin, pyrethrum, pyridaben, pyridathion, pyrimidifen, pyriproxyfen, quinalphos, ribavirin, salithion, sebufos, silafluofen, spinosad, spirodiclofen, sulfotep, sulprofos, tau-fluvalinate, tebufenozide, tebufenpyrad, tebupirimiphos, teflubenzuron, tefluthrin, temephos, temivinphos, terbufos, tetrachlorvinphos, tetradifon, theta-cypermethrin, thiacloprid, thiamethoxam, thiapronil, thiatriphos, thiocyclam hydrogen oxalate, thiodicarb, thiofanox, thuringiensin, tralocythrin, tralomethrin, triarathene, triazamate, triazophos, triazuron, trichlophenidine, trichlorfon, triflumuron, trimethacarb, vamidothion, vaniliprole, Verticillium lecanii, YI 5302, zeta-cypermethrin, zolaprofos, (1R-cis)-[5-(phenylmethyl)-3-furanyl]-methyl-3-[(dihydro-2-oxo-3(2H)-furanylidene)-methyl]-2,2-dimethylcyclopropanecarboxylate, (3-phenoxyphenyl)-methyl-2,2,3,3-tetramethylcyclopropanecarboxylate, 1-[(2-chloro-5-thiazolyl)methyl]tetrahydro-3,5-dimethyl-N-nitro-1,3,5-triazine-2(1H)-imine, 2-(2-chloro-6-fluorophenyl)4-[4-(1,1-dimethylethyl)phenyl]-4,5-dihydro-oxazole, 2-(acetyloxy)-3-dodecyl-1,4-naphthalenedione, 2-chloro-N-[[[4-(1-phenylethoxy)-phenyl]-amino]-carbonyl]-benzamide, 2-chloro-N-[[[4-(2,2-dichloro-1,1-difluoroethoxy)-phenyl]-amino]-carbonyl]-benzamide, 3-methylphenyl propylcarbamate, 4-[4-(4-ethoxyphenyl)-4-methylpentyl]-1-fluoro-2-phenoxy-benzene, 4-chloro-2-(1,1-dimethylethyl)-5-[[2-(2,6-dimethyl-4-phenoxyphenoxy)ethyl]thio]-3(2H)-pyridazinone, 4-chloro-2-(2-chloro-2-methylpropyl)-5-[(6-iodo-3-pyridinyl)methoxy]-3(2H)-pyridazinone, 4-chloro-5-[(6-chloro-3-pyridinyl)methoxy]-2-(3,4-dichlorophenyl)-3(2H)-pyridazinone, Bacillus thuringiensis strain EG-2348, [2-benzoyl-1-(1,1-dimethylethyl)-hydrazinobenzoic acid, 2,2-dimethyl-3-(2,4-dichlorophenyl)-2-oxo-1-oxaspiro[4.5]dec-3-en4-yl butanoate, [3-[(6-chloro-3-pyridinyl)methyl]-2-thiazolidinylidene]-cyanamide, dihydro-2-(nitromethylene)-2H-1,3-thiazine-3(4H)-carboxaldehyde, ethyl [2-[[1,6-dihydro-6-oxo-1-(phenylmethyl)-4-pyridazinyl]oxy]ethyl]-carbamate, N-(3,4,4-trifluoro-1-oxo-3-butenyl)-glycine, N-(4-chlorophenyl)-3-[4-(difluoromethoxy)phenyl]4,5-dihydro4-phenyl-1H-pyrazole-1-carboxamide, N-[(2-chloro-5-thiazolyl)methyl]-N′methyl-N″-nitro-guanidine, N-methyl-N′-(1-methyl-2-propenyl)-1,2-hydrazinedicarbothioamide, N-methyl-N′-2-propenyl-1,2-hydrazinedicarbothioamide, O,O-diethyl-[2-(dipropylamino)-2-oxoethyl]-ethylphosphoramidothioate, N-cyanomethyl-4-trifluoromethyl-nicotinamide, 3,5-dichloro-1-(3,3-dichloro-2-propenyloxy)-4-[3-(5-trifluoromethylpyridin-2-yloxy)-propoxy]-benzene.\",\n \"It is also possible to admix other known active compounds, such as herbicides, fertilizers and growth regulators.\",\n \"When used as insecticides, the active compounds according to the invention can furthermore be present in their commercially available formulations and in the use forms, prepared from these formulations, as a mixture with synergistic agents.\",\n \"Synergistic agents are compounds which increase the action of the active compounds, without it being necessary for the synergistic agent added to be active itself.\",\n \"The active compound content of the use forms prepared from the commercially available formulations can vary within wide limits.\",\n \"The active compound concentration of the use forms can be from 0.0000001 to 95% by weight of active compound, preferably between 0.0001 and 1% by weight.\",\n \"The compounds are employed in a customary manner appropriate for the use forms.\",\n \"When used against hygiene pests and pests of stored products, the active compound is distinguished by an excellent residual action on wood and clay as well as by a good stability to alkali on limed substrates.\",\n \"As already mentioned above, it is possible to treat all plants and their parts according to the invention.\",\n \"In a preferred embodiment, wild plant species and plant cultivars, or those obtained by conventional biological breeding, such as crossing or protoplast fusion, and parts thereof, are treated.\",\n \"In a further preferred embodiment, transgenic plants and plant cultivars obtained by genetic engineering, if appropriate in combination with conventional methods (Genetically Modified Organisms), and parts thereof are treated.\",\n \"The term “parts” or “parts of plants” or “plant parts” has been explained above.\",\n \"Particularly preferably, plants of the plant cultivars which are in each case commercially available or in use are treated according to the invention.\",\n \"Plant cultivars are understood as meaning plants with novel properties (“traits”) which are grown by conventional cultivation, by mutagenesis or by recombinant DNA techniques.\",\n \"These may be cultivars, biotypes or genotypes.\",\n \"Depending on the plant species or plant cultivars, their location and growth conditions (soils, climate, vegetation period, diet), the treatment according to the invention may also result in superadditive (“synergistic”) effects.\",\n \"Thus, for example, reduced application rates and/or a widening of the activity spectrum and/or an increase in the activity of the substances and compositions to be used according to the invention, better plant growth, increased tolerance to high or low temperatures, increased tolerance to drought or to water or soil salt content, increased flowering performance, easier harvesting, accelerated maturation, higher harvest yields, better quality and/or a higher nutritional value of the harvested products, better storage stability and/or processability of the harvested products are possible which exceed the effects which were actually to be expected.\",\n \"The transgenic plants or plant cultivars (i.e.\",\n \"those obtained by genetic engineering) which are preferably to be treated according to the invention include all plants which, in the genetic modification, received genetic material which imparted particularly advantageous useful properties (“traits”) to these plants.\",\n \"Examples of such properties are better plant growth, increased tolerance to high or low temperatures, increased tolerance to drought or to water or soil salt content, increased flowering performance, easier harvesting, accelerated maturation, higher harvest yields, better quality and/or a higher nutritional value of the harvested products, better storage stability and/or processability of the harvested products.\",\n \"Further and particularly emphasized examples of such properties are a better defence of the plants against animal and microbial pests, such as against insects, mites, phytopathogenic fungi, bacteria and/or viruses, and also increased tolerance of the plants to certain herbicidally active compounds.\",\n \"Examples of transgenic plants which may be mentioned are the important crop plants, such as cereals (wheat, rice), maize, soya beans, potatoes, cotton, oilseed rape and also fruit plants (with the fruits apples, pears, citrus fruits and grapes), and particular emphasis is given to maize, soya beans, potatoes, cotton and oilseed rape.\",\n \"Traits that are emphasized are in particular increased defence of the plants against insects by toxins formed in the plants, in particular those formed in the plants by the genetic material from Bacillus thuringiensis (for example by the genes CryIA(a), CryIA(b), CryIA(c), CryIIA, CryIIIA, CryIIIB2, Cry9c Cry2Ab, Cry3Bb and CryIF and also combinations thereof) (hereinbelow referred to as “Bt plants”).\",\n \"Traits which are also particularly emphasized are the increased resistance of plants to fungi, bacteria and viruses by systemic acquired resistance (SAR), systemin, phytoalexins, elicitors and resistance genes and the correspondingly expressed proteins and toxins.\",\n \"Traits that are furthermore particularly emphasized are the increased tolerance of the plants to certain herbicidally active compounds, for example imidazolinones, sulphonylureas, glyphosate or phosphinotricin (for example the “PAT” gene).\",\n \"The genes which impart the desired traits in question can also be present in combination with one another in the transgenic plants.\",\n \"Examples of “Bt plants” which may be mentioned are maize varieties, cotton varieties, soya bean varieties and potato varieties which are sold under the trade names YIELD GARD® (for example maize, cotton, soya beans), KnockOut® (for example maize), StarLink® (for example maize), Bollgard® (cotton), Nucotn® (cotton) and NewLeaf® (potato).\",\n \"Examples of herbicide-tolerant plants which may be mentioned are maize varieties, cotton varieties and soya bean varieties which are sold under the trade names Roundup Ready® (tolerance to glyphosate, for example maize, cotton, soya bean), Liberty Link® (tolerance to phosphinotricin, for example oilseed rape), IMI® (tolerance to imidazolinones) and STS® (tolerance to sulphonylureas, for example maize).\",\n \"Herbicide-resistant plants (plants bred in a conventional manner for herbicide tolerance) which may be mentioned include the varieties sold under the name Clearfield® (for example maize).\",\n \"Of course, these statements also apply to plant cultivars having these genetic traits or genetic traits still to be developed, which cultivars will be developed and/or marketed in the future.\",\n \"The plants listed can be treated according to the invention in a particularly advantageous manner with the compounds of the formula I or the active compound mixtures according to the invention.\",\n \"The preferred ranges stated above for the active compounds or mixtures also apply to the treatment of these plants.\",\n \"Particular emphasis is given to the treatment of plants with the compounds or mixtures specifically mentioned in the present text.\",\n \"The active compounds according to the invention act not only against plant, hygiene and stored product pests, but also in the veterinary medicine sector against animal parasites (ectoparasites), such as hard ticks, soft ticks, mange mites, leaf mites, flies (biting and licking), parasitic fly larvae, lice, hair lice, feather lice and fleas.\",\n \"These parasites include: From the order of the Anoplurida, for example, Haematopinus spp., Linognathus spp., Pediculus spp., Phtirus spp.\",\n \"and Solenopotes spp.\",\n \"From the order of the Mallophagida and the suborders Amblycerina and Ischnocerina, for example, Trimenopon spp., Menopon spp., Trinoton spp., Bovicola spp., Werneckiella spp., Lepikentron spp., Damalina spp., Trichodectes spp.\",\n \"and Felicola spp.\",\n \"From the order of the Diptera and the suborders Nematocerina and Brachycerina, for example, Aedes spp., Anopheles spp., Culex spp., Simulium spp., Eusimulium spp., Phlebotomus spp., Lutzomyia spp., Culicoides spp., Chrysops spp., Hybomitra spp., Atylotus spp., Tabanus spp., Haematopota spp., Philipomyia spp., Braula spp., Musca spp., Hydrotaea spp., Stomoxys spp., Haematobia spp., Morellia spp., Fannia spp., Glossina spp., Calliphora spp., Lucilia spp., Chrysomyia spp., Wohlfahrtia spp., Sarcophaga spp., Oestrus spp., Hypoderma spp., Gasterophilus spp., Hippobosca spp., Lipoptena spp.\",\n \"and Melophagus spp.\",\n \"From the order of the Siphonapterida, for example Pulex spp., Ctenocephalides spp., Xenopsylla spp.\",\n \"and Ceratophyllus spp.\",\n \"From the order of the Heteropterida, for example, Cimex spp., Triatoma spp., Rhodnius spp.\",\n \"and Panstrongylus spp.\",\n \"From the order of the Blattarida, for example Blatta orientalis, Periplaneta americana, Blattela germanica and Supella spp.\",\n \"From the subclass of the Acaria (Acarida) and the orders of the Meta- and Mesostigmata, for example, Argas spp., Ornithodorus spp., Otobius spp., Ixodes spp., Amblyomma spp., Boophilus spp., Dermacentor spp., Haemophysalis spp., Hyalomma spp., Rhipicephalus spp., Dermanyssus spp., Raillietia spp., Pneumonyssus spp., Sternostoma spp.\",\n \"and Varroa spp.\",\n \"From the order of the Actinedida (Prostigmata) und Acaridida (Astigmata), for example, Acarapis spp., Cheyletiella spp., Ornithocheyletia spp., Myobia spp., Psorergates spp., Demodex spp., Trombicula spp., Listrophorus spp., Acarus spp., Tyrophagus spp., Caloglyphus spp., Hypodectes spp., Pterolichus spp., Psoroptes spp., Chorioptes spp., Otodectes spp., Sarcoptes spp., Notoedres spp., Knemidocoptes spp., Cytodites spp.\",\n \"and Laminosioptes spp.\",\n \"The active compounds of the formula (I) according to the invention are also suitable for controlling arthropods which infest agricultural productive livestock, such as, for example, cattle, sheep, goats, horses, pigs, donkeys, camels, buffalo, rabbits, chickens, turkeys, ducks, geese and bees, other pets, such as, for example, dogs, cats, caged birds and aquarium fish, and also so-called test animals, such as, for example, hamsters, guinea pigs, rats and mice.\",\n \"By controlling these arthropods, cases of death and reduction in productivity (for meat, milk, wool, hides, eggs, honey etc.)\",\n \"should be diminished, so that more economic and easier animal husbandry is possible by use of the active compounds according to the invention.\",\n \"The active compounds according to the invention are used in the veterinary sector in a known manner by enteral administration in the form of, for example, tablets, capsules, potions, drenches, granules, pastes, boluses, the feed-through process and suppositories, by parenteral administration, such as, for example, by injection (intramuscular, subcutaneous, intravenous, intraperitoneal and the like), implants, by nasal administration, by dermal use in the form, for example, of dipping or bathing, spraying, pouring on and spotting on, washing and powdering, and also with the aid of molded articles containing the active compound, such as collars, ear marks, tail marks, limb bands, halters, marking devices and the like.\",\n \"When used for cattle, poultry, pets and the like, the active compounds of the formula (I) can be used as formnulations (for example powders, emulsions, free-flowing compositions), which comprise the active compounds in an amount of 1 to 80% by weight, directly or after 100 to 10 000-fold dilution, or they can be used as a chemical bath.\",\n \"It has furthermore been found that the compounds according to the invention have a strong insecticidal action against insects which destroy industrial materials.\",\n \"The following insects may be mentioned as examples and as preferred—but without a limitation: Beetles, such as Hylotrupes bajulus, Chlorophorus pilosis, Anobium punctatum, Xestobium rufovillosum, Ptilinus pecticomis, Dendrobium pertinex, Ernobius mollis, Priobium carpini, Lyctus brunneus, Lyctus africanus, Lyctus planicollis, Lyctus linearis, Lyctus pubescens, Trogoxylon aequale, Minthes rugicollis, Xyleborus spec.\",\n \"Tryptodendron spec.\",\n \"Apate monachus, Bostrychus capucins, Heterobostrychus brunneus, Sinoxylon spec.\",\n \"Dinoderus minutus.\",\n \"Hymenopterons, such as Sirex juvencus, Urocerus gigas, Urocerus gigas taignus, Urocerus augur.\",\n \"Termites, such as Kalotermes flavicollis, Cryptotermes brevis, Heterotermes indicola, Reticulitermes flavipes, Reticulitermes santonensis, Reticulitermes lucifugus, Mastotermes darwiniensis, Zootermopsis nevadensis, Coptotermes formosanus.\",\n \"Bristletails, such as Lepisma saccharina.\",\n \"Industrial materials in the present connection are to be understood as meaning non-living materials, such as, preferably, plastics, adhesives, sizes, papers and cards, leather, wood and processed wood products and coating compositions.\",\n \"Wood and processed wood products are materials to be protected, especially preferably, from insect infestation.\",\n \"Wood and processed wood products which can be protected by the agents according to the invention or mixtures comprising these are to be understood as meaning, for example: building timber, wooden beams, railway sleepers, bridge components, boat jetties, wooden vehicles, boxes, pallets, containers, telegraph poles, wood panelling, wooden windows and doors, plywood, chipboard, joinery or wooden products which are used quite generally in house-building or in building joinery.\",\n \"The active compounds can be used as such; in the form of concentrates or generally customary formulations, such as powders, granules, solutions, suspensions, emulsions or pastes.\",\n \"The formulations mentioned can be prepared in a manner known per se, for example by mixing the active compounds with at least one solvent or diluent, emulsifier, dispersing agent and/or binder or fixing agent, a water repellent, if appropriate siccatives and UV stabilizers and if appropriate dyestuffs and pigments, and also other processing auxiliaries.\",\n \"The insecticidal compositions or concentrates used for the preservation of wood and wood-derived timber products comprise the active compound according to the invention in a concentration of 0.0001 to 95% by weight, in particular 0.001 to 60% by weight.\",\n \"The amount of the compositions or concentrates employed depends on the nature and occurrence of the insects and on the medium.\",\n \"The optimum amount employed can be determined for the use in each case by a series of tests.\",\n \"In general, however, it is sufficient to employ 0.0001 to 20% by weight, preferably 0.001 to 10% by weight, of the active compound, based on the material to be preserved.\",\n \"Solvents and/or diluents which are used are an organic chemical solvent or solvent mixture and/or an oily or oil-like organic chemical solvent or solvent mixture of low volatility and/or a polar organic chemical solvent or solvent mixture and/or water, and if appropriate an emulsifier and/or wetting agent.\",\n \"Organic chemical solvents which are preferably used are oily or oil-like solvents having an evaporation number above 35 and a flashpoint above 30° C., preferably above 45° C. Substances which are used as such oily or oil-like water-insoluble solvents of low volatility are appropriate mineral oils or aromatic fractions thereof, or solvent mixtures containing mineral oils, preferably white spirit, petroleum and/or alkylbenzene.\",\n \"Mineral oils having a boiling range from 170 to 220° C., white spirit having a boiling range from 170 to 220° C., spindle oil having a boiling range from 250 to 350° C., petroleum and aromatics having a boiling range from 160 to 280° C., terpentine oil and the like, are advantageously employed.\",\n \"In a preferred embodiment, liquid aliphatic hydrocarbons having a boiling range from 180 to 210° C. or high-boiling mixtures of aromatic and aliphatic hydrocarbons having a boiling range from 180 to 220° C. and/or spindle oil and/or monochloronaphthalene, preferably α-monochloronaphthalene, are used.\",\n \"The organic oily or oil-like solvents of low volatility which have an evaporation number above 35 and a flashpoint above 30° C., preferably above 45° C., can be replaced in part by organic chemical solvents of high or medium volatility, providing that the solvent mixture likewise has an evaporation number above 35 and a flashpoint above 30° C., preferably above 45° C., and that the insecticide/fungicide mixture is soluble or emulsifiable in this solvent mixture.\",\n \"According to a preferred embodiment, some of the organic chemical solvent or solvent mixture or an aliphatic polar organic chemical solvent or solvent mixture is replaced.\",\n \"Aliphatic organic chemical solvents containing hydroxyl and/or ester and/or ether groups, such as, for example, glycol ethers, esters or the like, are preferably used.\",\n \"Organic chemical binders which are used in the context of the present invention are the synthetic resins and/or binding drying oils which are known per se, are water-dilutable and/or are soluble or dispersible or emulsifiable in the organic chemical solvents employed, in particular binders consisting of or comprising an acrylate resin, a vinyl resin, for example polyvinyl acetate, polyester resin, polycondensation or polyaddition resin, polyurethane resin, alkyd resin or modified alkyd resin, phenolic resin, hydrocarbon resin, such as indene-cumarone resin, silicone resin, drying vegetable oils and/or drying oils and/or physically drying binders based on a natural and/or synthetic resin.\",\n \"The synthetic resin used as the binder can be employed in the form of an emulsion, dispersion or solution.\",\n \"Bitumen or bituminous substances can also be used as binders in an amount of up to 10% by weight.\",\n \"Dyestuffs, pigments, water-repelling.\",\n \"agents, odour correctants and inhibitors or anticorrosive agents and the like which are known per se can additionally be employed.\",\n \"It is preferred according to the invention for the composition or concentrate to comprise, as the organic chemical binder, at least one alkyd resin or modified alkyd resin and/or one drying vegetable oil.\",\n \"Alkyd resins having an oil content of more than 45% by weight, preferably 50 to 68% by weight, are preferably used according to the invention.\",\n \"All or some of the binder mentioned can be replaced by a fixing agent (mixture) or a plasticizer (mixture).\",\n \"These additives are intended to prevent evaporation of the active compounds and crystallization or precipitation.\",\n \"They preferably replace 0.01 to 30% of the binder (based on 100% of the binder employed).\",\n \"The plasticizers originate from the chemical classes of phthalic acid esters, such as dibutyl, dioctyl or benzyl butyl phthalate, phosphoric acid esters, such as tributyl phosphate, adipic acid esters, such as di-(2-ethylhexyl) adipate, stearates, such as butyl stearate or amyl stearate, oleates, such as butyl oleate, glycerol ethers or higher molecular weight glycol ethers, glycerol esters and p-toluenesulphonic acid esters.\",\n \"Fixing agents are based chemically on polyvinyl alkyl ethers, such as, for example, polyvinyl methyl ether or ketones, such as benzophenone or ethylenebenzophenone.\",\n \"Possible solvents or diluents are, in particular, also water, if appropriate as a mixture with one or more of the abovementioned organic chemical solvents or diluents, emulsifiers and dispersing agents.\",\n \"Particularly effective preservation of wood is achieved by impregnation processes on a large industrial scale, for example vacuum, double vacuum or pressure processes.\",\n \"The, ready-to-use compositions can also comprise other insecticides, if appropriate, and also one or more fungicides, if appropriate.\",\n \"Possible additional mixing partners are, preferably, the insecticides and fungicides mentioned in WO 94/29 268.The compounds mentioned in this document are an explicit constituent of the present application.\",\n \"Especially preferred mixing partners which may be mentioned are insecticides, such as chlorpyriphos, phoxim, silafluofin, alphamethrin, cyfluthrin, cypermethrin, deltamethrin, permethrin, imidacloprid, NI-25, flufenoxuron, hexaflumuron, transfluthrin, thiacloprid, methoxyfenozide and triflumuron, and also fungicides, such as epoxyconazole, hexaconazole, azaconazole, propiconazole, tebuconazole, cyproconazole, metconazole, imazalil, dichlorfluanid, tolylfluanid, 3-iodo-2-propinyl-butyl carbamate, N-octyl-isothiazolin-3-one and 4,5-dichloro-N-octylisothiazolin-3-one.\",\n \"The compounds according to the invention can at the same time be employed for protecting objects which come into contact with saltwater or brackish water, such as hulls, screens, nets, buildings, moorings and signalling systems, against fouling.\",\n \"Fouling by sessile Oligochaeta, such as Serpulidae, and by shells and species from the Ledamorpha group (goose barnacles), such as various Lepas and Scalpellum species, or by species from the Balanomorpha group (acorn barnacles), such as Balanus or Pollicipes species, increases the frictional drag of ships and, as a consequence, leads to a marked increase in operation costs owing to higher energy consumption and additionally frequent residence in the dry dock.\",\n \"Apart from fouling by algae, for example Ectocarpus sp.\",\n \"and Ceramium sp., in particular fouling by sessile Entomostraka groups, which come under the generic term Cirripedia (cirriped crustaceans), is of particular importance.\",\n \"Surprisingly, it has now been found that the compounds according to the invention, alone or in combination with other active compounds, have an outstanding antifouling action.\",\n \"Using the compounds according to the invention, alone or in combination with other active compounds, allows the use of heavy metals such as, for example, in bis(trialkyltin) sulphides, tri-n-butyltin laurate, tri-n-butyltin chloride, copper(I) oxide, triethyltin chloride, tri-n-butyl(2-phenyl4-chlorophenoxy)tin, tributyltin oxide, molybdenum disulphide, antimony oxide, polymeric butyl titanate, phenyl-(bispyridine)bismuth chloride, tri-n-butyltin fluoride, manganese ethylenebis-thiocarbamate, zinc dimethyldithiocarbamate, zinc ethylenebisthiocarbamate, zinc salts and copper salts of 2-pyridinethiol 1-oxide, bisdimethyldithiocarbamoylzinc ethylene-bisthiocarbamate, zinc oxide, copper(I) ethylene-bisdithiocarbamate, copper thiocyanate, copper naphthenate and tributyltin halides to be dispensed with, or the concentration of these compounds to be substantially reduced.\",\n \"If appropriate, the ready-to-use antifouling paints can additionally comprise other active compounds, preferably algicides, fungicides, herbicides, molluscicides, or other antifouling active compounds.\",\n \"Preferably suitable components in combinations with the antifouling compositions according to the invention are: algicides such as 2-tert-butylamino-4-cyclopropylamino-6-methylthio-1,3,5-triazine, dichlorophen, diuron, endothal, fentin acetate, isoproturon, methabenzthiazuron, oxyfluorfen, quinoclamine and terbutryn; fungicides such as benzo[b]thiophenecarboxylic acid cyclohexylamide S,S-dioxide, dichlofluanid, fluorfolpet, 3-iodo-2-propinyl butylcarbamate, tolylfluanid and azoles such as azaconazole, cyproconazole, epoxyconazole, hexaconazole, metconazole, propiconazole and tebuconazole; molluscicides such as fentin acetate, metaldehyde, methiocarb, niclosamid, thiodicarb and trimethacarb; or conventional antifouling active compounds such as 4,5-dichloro-2-octyl-4-isothiazolin-3-one, diiodomethylparatryl sulphone, 2-(N,N-dimethylthiocarbamoylthio)-5-nitrothiazyl, potassium, copper, sodium and zinc salts of 2-pyridinethiol 1-oxide, pyridine-triphenylborane, tetrabutyldistannoxane, 2,3,5,6-tetrachloro-4-(methylsulphonyl)-pyridine, 2,4,5,6-tetrachloroisophthalonitrile, tetramethylthiuram disulphide and 2,4,6-trichlorophenylmaleiimide.\",\n \"The antifouling compositions used comprise the active compound according to the invention of the compounds according to the invention in a concentration of 0.001 to 50% by weight, in particular 0.01 to 20% by weight.\",\n \"Moreover, the antifouling compositions according to the invention comprise the customary components such as, for example, those described in Ungerer, Chem.\",\n \"Ind.\",\n \"1985, 37, 730-732 and Williams, Antifouling Marine Coatings, Noyes, Park Ridge, 1973.Besides the algicidal, fungicidal, molluscicidal active compounds and insecticidal active compounds according to the invention, antifouling paints comprise, in particular, binders.\",\n \"Examples of recognized binders are polyvinyl chloride in a solvent system, chlorinated rubber in a solvent system, acrylic resins in a solvent system, in particular in an aqueous system, vinyl chloride/vinyl acetate copolymer systems in the form of aqueous dispersions or in the form of organic solvent systems, butadiene/styrene/acrylonitrile rubbers, drying oils such as linseed oil, resin esters or modified hardened resins in combination with tar or bitumens, asphalt and epoxy compounds, small amounts of chlorine rubber, chlorinated polypropylene and vinyl resins.\",\n \"If appropriate, paints also comprise inorganic pigments, organic pigments or colorants which are preferably insoluble in salt water.\",\n \"Paints may furthermore comprise materials such as colophonium to allow controlled release of the active compounds.\",\n \"Furthermore, the paints may comprise plasticizers, modifiers which affect the rheological properties and other conventional constituents.\",\n \"The compounds according to the invention or the abovementioned mixtures may also be incorporated into self-polishing antifouling systems.\",\n \"The active compounds are also suitable for controlling animal pests, in particular insects, arachnids and mites, which are found in enclosed spaces such as, for example, dwellings, factory halls, offices, vehicle cabins and the like.\",\n \"They can be employed alone or in combination with other active compounds and auxiliaries in domestic insecticide products for controlling these pests.\",\n \"They are active against sensitive and resistant species and against all developmental stages.\",\n \"These pests include: From the order of the Scorpionidea, for example, Buthus occitanus.\",\n \"From the order of the Acarina, for example, Argas persicus, Argas reflexus, Bryobia spp., Dermanyssus gallinae, Glyciphagus domesticus, Ornithodorus moubat, Rhipicephalus sanguineus, Trombicula alfreddugesi, Neutrombicula autumnalis, Dermatophagoides pteronissimus, Dermatophagoides forinae.\",\n \"From the order of the Araneae, for example, Aviculariidae, Araneidae.\",\n \"From the order of the Opiliones, for example, Pseudoscorpiones chelifer, Pseudoscorpiones cheiridium, Opiliones phalangium.\",\n \"From the order of the Isopoda, for example, Oniscus asellus, Porcellio scaber.\",\n \"From the order of the Diplopoda, for example, Blaniulus guttulatus, Polydesmus spp.\",\n \"From the order of the Chilopoda, for example, Geophilus spp.\",\n \"From the order of the Zygentoma, for example, Ctenolepisma spp., Lepisma saccharina, Lepismodes inquilinus.\",\n \"From the order of the Blattaria, for example, Blatta orientalies, Blattella germanica, Blattella asahinai, Leucophaea maderae, Panchlora spp., Parcoblatta spp., Periplaneta australasiae, Periplaneta americana, Periplaneta brunnea, Periplaneta fuliginosa, Supella longipalpa.\",\n \"From the order of the Saltatoria, for example, Acheta domesticus.\",\n \"From the order of the Dermaptera, for example, Forficula auricularia.\",\n \"From the order of the Isoptera, for example, Kalotermes spp., Reticulitermes spp.\",\n \"From the order of the Psocoptera, for example, Lepinatus spp., Liposcelis spp.\",\n \"From the order of the Coleptera, for example, Anthrenus spp., Attagenus spp., Dermestes spp., Latheticus oryzae, Necrobia spp., Ptinus spp., Rhizopertha dominica, Sitophilus granarius, Sitophilus oryzae, Sitophilus zeamais, Stegobium paniceum.\",\n \"From the order of the Diptera, for example, Aedes aegypti, Aedes albopictus, Aedes taeniorhynchus, Anopheles spp., Calliphora erythrocephala, Chrysozona pluvialis, Culex quinquefasciatus, Culex pipiens, Culex tarsalis, Drosophila spp., Fannia canicularis, Musca domestica, Phlebotomus spp., Sarcophaga carnaria, Simulium spp., Stomoxys calcitrans, Tipula paludosa.\",\n \"From the order of the Lepidoptera, for example, Achroia grisella, Galleria mellonella, Plodia interpunctella, Tinea cloacella, Tinea pellionella, Tineola bisselliella.\",\n \"From the order of the Siphonaptera, for example, Ctenocephalides canis, Ctenocephalides felis, Pulex irritans, Tunga penetrans, Xenopsylla cheopis.\",\n \"From the order of the Hymenoptera, for example, Camponotus herculeanus, Lasius fuliginosus, Lasius niger, Lasius umbratus, Monomorium pharaonis, Paravespula spp., Tetramorium caespitum.\",\n \"From the order of the Anoplura, for example, Pediculus humanus capitis, Pediculus humanus corporis, Phthirus pubis.\",\n \"From the order of the Heteroptera, for example, Cimex hemipterus, Cimex lectularius, Rhodinus prolixus, Triatoma infestans.\",\n \"In the field of household insecticides, they are used alone or in combination with other suitable active compounds, such as phosphoric acid esters, carbamates, pyrethroids, growth regulators or active compounds from other known classes of insecticides.\",\n \"They are used as aerosols, pressure-free spray products, for example pump and atomizer sprays, automatic fogging systems, foggers, foams, gels, evaporator products with evaporator tablets made of cellulose or polymer, liquid evaporators, gel and membrane evaporators, propeller-driven evaporators, energy-free or passive, evaporation systems, moth papers, moth bags and moth gels, as granules or dusts, in baits for spreading or in bait stations.\",\n \"The active compounds according to the invention can also be used as defoliants, desiccants, haulm killers and, especially, as weedkillers.\",\n \"Weeds in the broadest sense are understood to mean all plants which grow in locations where they are undesired.\",\n \"Whether the substances according to the invention act as total or selective herbicides depends essentially on the amount used.\",\n \"The active compounds according to the invention can be used, for example, in connection with the following plants: Dicotyledonous weeds of the genera: Abutilon, Amaranthus, Ambrosia, Anoda, Anthemis, Aphanes, Atriplex, Bellis, Bidens, Capsella, Carduus, Cassia, Centaurea, Chenopodium, Cirsium, Convolvulus, Datura, Desmodium, Emex, Erysimum, Euphorbia, Galeopsis, Galinsoga, Galium, Hibiscus, Ipomoea, Kochia, Lamium, Lepidium, Lindernia, Matricaria, Mentha, Mercurialis, Mullugo, Myosotis, Papaver, Pharbitis, Plantago, Polygonum, Portulaca, Ranunculus, Raphanus, Rorippa, Rotala, Rumex, Salsola, Senecio, Sesbania, Sida, Sinapis, Solanum, Sonchus, Sphenoclea, Stellaria, Taraxacum, Thlaspi, Trifolium, Urtica, Veronica, Viola, Xanthium.\",\n \"Dicotyledonous crops of the genera: Arachis, Beta, Brassica, Cucumis, Cucurbita, Helianthus, Daucus, Glycine, Gossypium, Ipomoea, Lactuca, Linum, Lycopersicon, Nicotiana, Phaseolus, Pisum, Solanum, Vicia.\",\n \"Monocotyledonous weeds of the genera: Aegilops, Agropyron, Agrostis, Alopecurus, Apera, Avena, Brachiaria, Bromus, Cenchrus, Commelina, Cynodon, Cyperus, Dactyloctenium, Digitaria, Echinochloa, Eleocharis, Eleusine, Eragrostis, Eriochloa, Festuca, Fimbristylis, Heteranthera, Imperata, Ischaemum, Leptochloa, Lolium, Monochoria, Panicum, Paspalum, Phalaris, Phleum, Poa, Rottboellia, Sagittaria, Scirpus, Setaria, Sorghum.\",\n \"Monocotyledonous crops of the genera: Allium, Ananas, Asparagus, Avena, Hordeum, Oryza, Panicum, Saccharum, Secale, Sorghum, Triticale, Triticum, Zea.\",\n \"However, the use of the active compounds according to the invention is in no way restricted to these genera, but also extends in the same manner to other plants.\",\n \"The active compounds according to the invention are suitable, depending on the concentration, for the total control of weeds, for example on industrial terrain and rail tracks, and on paths and areas with and without tree plantings.\",\n \"Similarly, the active compounds according to the invention can be employed for controlling weeds in perennial crops, for example forests, decorative tree plantings, orchards, vineyards, citrus groves, nut orchards, banana plantations, coffee plantations, tea plantations, rubber plantations, oil palm plantations, cocoa plantations, soft fruit plantings and hop fields, on lawns, turf and pastureland, and for the selective control of weeds in annual crops.\",\n \"The compounds of the formula (I) according to the invention have strong herbicidal activity and a broad active spectrum when used on the soil and on above-ground parts of plants.\",\n \"To a certain extent they are also suitable for the selective control of monocotyledonous and dicotyledonous weeds in monocotyledonous and dicotyledonous crops, both by the pre-emergence and by the post-emergence method.\",\n \"At certain concentrations or application rates, the active compounds according to the invention can also be employed for controlling animal pests and fungal or bacterial plant diseases.\",\n \"If appropriate, they can also be used as intermediates or precursors for the synthesis of other active compounds.\",\n \"The active compounds can be converted into the customary formulations such as solutions, emulsions, wettable powders, suspensions, powders, dusts, pastes, soluble powders, granules, suspension-emulsion concentrates, natural and synthetic materials impregnated with active compound, and microencapsulations in polymeric materials.\",\n \"These formulations are produced in a known manner, for example by mixing the active compounds with extenders, that is, liquid solvents and/or solid carriers, optionally with the use of surfactants, that is, emulsifiers and/or dispersants, and/or foam formers.\",\n \"If the extender used is water, it is also possible to employ, for example, organic solvents as auxiliary solvents.\",\n \"Essentially, suitable liquid solvents are: aromatics such as xylene, toluene or alkylnaphthalenes, chlorinated aromatics and chlorinated aliphatic hydrocarbons such as chlorobenzenes, chloroethylenes or methylene chloride, aliphatic hydrocarbons such as cyclohexane or paraffins, for example mineral oil fractions, mineral and vegetable oils, alcohols such as butanol or glycol and their ethers and esters, ketones such as acetone, methyl ethyl ketone, methyl isobutyl ketone or cyclohexanone, strongly polar solvents such as dimethylformamide and dimethyl sulphoxide, or else water.\",\n \"Suitable solid carriers are: for example ammonium salts and ground natural minerals such as kaolins, clays, talc, chalk, quartz, attapulgite, montmorillonite or diatomaceous earth, and ground synthetic materials such as finely divided silica, alumina and silicates; suitable solid carriers for granules are: for example crushed and fractionated natural rocks such as calcite, marble, pumice, sepiolite and dolomite, or else synthetic granules of inorganic and organic meals, and granules of organic material such as sawdust, coconut shells, maize cobs and tobacco stalks; suitable emulsifiers and/or foam formers are: for example nonionic and anionic emulsifiers such as polyoxyethylene fatty acid esters, polyoxyethylene fatty alcohol ethers, for example alkylaryl polyglycol ethers, alkylsulphonates, alkyl sulphates, arylsulphonates, or else protein hydrolysates; suitable dispersants are: for example lignosulphite waste liquors and methylcellulose.\",\n \"Tackifiers such as carboxymethylcellulose and natural and synthetic polymers in the form of powders, granules or latices, such as gum arabic, polyvinyl alcohol and polyvinyl acetate, or else natural phospholipids such as cephalins and lecithins and synthetic phospholipids can be used in the formulations.\",\n \"Other possible additives are mineral and vegetable oils.\",\n \"It is possible to use colorants such as inorganic pigments, for example iron oxide, titanium oxide and Prussian Blue, and organic colorants such as alizarin colorants, azo colorants and metal phthalocyanine colorants, and trace nutrients such as salts of iron, manganese, boron, copper, cobalt, molybdenum and zinc.\",\n \"The formulations generally comprise between 0.1 and 95% by weight of active compound, preferably between 0.5 and 90%.\",\n \"For controlling weeds, the active compounds according to the invention, as such or in their formulations, can also be used as mixtures with known herbicides and/or substances which improve the compatibility with crop plants (“safeners”), finished formulations or tank mixes being possible.\",\n \"Also possible are mixtures with weed-killers comprising one or more known herbicides and a safener.\",\n \"Possible components for the mixtures are known herbicides, for example acetochlor, acifluorfen (-sodium), aclonifen, alachlor, alloxydim (-sodium), ametryne, amicarbazone, amidochlor, amidosulfuron, anilofos, asulam, atrazine, azafenidin, azimsulfuron, beflubutamid, benazolin (-ethyl), benfuresate, bensulfuron (-methyl), bentazon, benzfendizone, benzobicyclon, benzofenap, benzoylprop (-ethyl), bialaphos, bifenox, bispyribac (-sodium), bromobutide, bromofenoxim, bromoxynil, butachlor, butafenacil (-allyl), butroxydim, butylate, cafenstrole, caloxydim, carbetamide, carfentrazone (-ethyl), chlomethoxyfen, chloramben, chloridazon, chlorimuron (-ethyl), chlornitrofen, chlorsulfuron, chlortoluron, cinidon (-ethyl), cinmethylin, cinosulfuron, clefoxydim, clethodim, clodinafop (-propargyl), clomazone, clomeprop, clopyralid, clopyrasulfuron (-methyl), cloransulam (-methyl), cumyluron, cyanazine, cybutryne, cycloate, cyclosulfamuron, cycloxydim, cyhalofop (-butyl), 2,4-D, 2,4-DB, desmedipham, diallate, dicamba, dichlorprop (—P), diclofop (-methyl), diclosulam, diethatyl (-ethyl), difenzoquat, diflufenican, diflufenzopyr, dimefuron, dimepiperate, dimethachlor, dimethametryn, dimethenamid, dimexyflam, dinitramine, diphenamid, diquat, dithiopyr, diuron, dymron, epropodan, EPTC, esprocarb, ethalfluralin, ethametsulfuron (-methyl), ethofumesate, ethoxyfen, ethoxysulfuron, etobenzanid, fenoxaprop (—P-ethyl), fentrazamide, flamprop (-isopropyl, -isopropyl-L, -methyl), flazasulfuron, florasulam, fluazifop (—P-butyl), fluazolate, flucarbazone (-sodium), flufenacet, flumetsulam, flumiclorac (-pentyl), flumioxazin, flumipropyn, flumetsulam, fluometuron, fluorochloridone, fluoroglycofen (-ethyl), flupoxam, flupropacil, flurpyrsulfuron (-methyl, -sodium), flurenol (-butyl), fluridone, fluroxypyr (-butoxypropyl, -meptyl), flurprimidol, flurtamone, fluthiacet (-methyl), fluthiamide, fomesafen, foramsulfuron, glufosinate (-ammonium), glyphosate (-isopropylammonium), halosafen, haloxyfop (-ethoxyethyl, —P-methyl), hexazinone, imazamethabenz (-methyl), imazamethapyr, imazamox, imazapic, imazapyr, imazaquin, imazethapyr, imazosulfuron, iodosulfuron (-methyl, -sodium), ioxynil, isopropalin, isoproturon, isouron, isoxaben, isoxachlortole, isoxaflutole, isoxapyrifop, lactofen, lenacil, linuron, MCPA, mecoprop, mefenacet, mesotrione, metamitron, metazachlor, methabenzthiazuron, metobenzuron, metobromuron, (alpha-) metolachlor, metosulam, metoxuron, metribuzin, metsulfuron (-methyl), molinate, monolinuron, naproanilide, napropamide, neburon, nicosulfuron, norflurazon, orbencarb, oryzalin, oxadiargyl, oxadiazon, oxasulfuron, oxaziclomefone, oxyfluorfen, paraquat, pelargonic acid, pendimethalin, pendralin, pentoxazone, phenmedipham, picolinafen, piperophos, pretilachlor, primisulfuron (-methyl), profluazol, prometryn, propachlor, propanil, propaquizafop, propisochlor, propoxycarbazone (-sodium), propyzamide, prosulfocarb, prosulfuron, pyraflufen (-ethyl), pyrazogyl, pyrazolate, pyrazosulfuron (-ethyl), pyrazoxyfen, pyribenzoxim, pyributicarb, pyridate, pyridatol, pyriftalid, pyriminobac (-methyl), pyrithiobac (-sodium), quinchlorac, quinmerac, quinoclamine, quizalofop (—P-ethyl, —P-tefuryl), rimsulfuron, sethoxydim, simazine, simetryn, sulcotrione, sulfentrazone, sulfometuron (-methyl), sulfosate, sulfosulfuron, tebutam, tebuthiuron, tepraloxydim, terbuthylazine, terbutryn, thenylchlor, thiafluamide, thiazopyr, thidiazimin, thifensulfuron (-methyl), thiobencarb, tiocarbazil, tralkoxydim, tniallate, triasulfuron, tribenuron (-methyl), triclopyr, tridiphane, trifluralin, trifloxysulfuron, triflusulfuron (-methyl), tritosulfuron.\",\n \"Furthermore suitable for the mixtures are known safeners, for example: AD-67, BAS-145138, benoxacor, cloquintocet (-mexyl), cyometrinil, 2,4-D, DKA-24, dichlormid, dymron, fenclorim, fenchlorazol (-ethyl), flurazole, fluxofenim, furilazole, isoxadifen (-ethyl), MCPA, mecoprop (—P), mefenpyr (-diethyl), MG-191, oxabetrinil, PPG-1292, R-29148.A mixture with other known active compounds, such as fungicides, insecticides, acaricides, nematicides, bird repellents, plant nutrients and agents which improve soil structure, is also possible.\",\n \"The active compounds can be used as such, in the form of their formulations or in the use forms prepared therefrom by further dilution, such as ready-to-use solutions, suspensions, emulsions, powders, pastes and granules.\",\n \"They are used in a customary manner, for example by watering, spraying, atomizing or broadcasting.\",\n \"The active compounds according to the invention can be applied both before and after emergence of the plants.\",\n \"They can also be incorporated into the soil before sowing.\",\n \"The amount of active compound used can vary within a relatively wide range.\",\n \"It depends essentially on the nature of the desired effect.\",\n \"In general, the amounts used are between 1 g and 10 kg of active compound per hectare of soil surface, preferably between 5 g and 5 kg per ha.\",\n \"The substances according to the invention have potent microbicidal activity and can be employed for controlling undesirable microorganisms, such as fungi and bacteria, in crop protection and in the protection of materials.\",\n \"Fungicides can be employed in crop protection for controlling Plasmodiophoromycetes, Oomycetes, Chytridiomycetes, Zygomycetes, Ascomycetes, Basidiomycetes and Deuteromycetes.\",\n \"Bactericides can be employed in crop protection for controlling Pseudomonadaceae, Rhizobiaceae, Enterobacteriaceae, Corynebacteriaceae and Streptomycetaceae.\",\n \"Some pathogens causing fungal and bacterial diseases which come under the generic names listed above may be mentioned as examples, but not by way of limitation: Xanthomonas species, such as, for example, Xanthomonas campestris pv.\",\n \"oryzae; Pseudomonas species, such as, for example, Pseudomonas syringae pv.\",\n \"lachrymans; Erwinia species, such as, for example, Erwinia amylovora; Pythium species, such as, for example, Pythium ultimum; Phytophthora species, such as, for example, Phytophthora infestans; Pseudoperonospora species, such as, for example, Pseudoperonospora humuli or Pseudoperonospora cubensis; Plasmopara species, such as, for example, Plasmopara viticola; Bremia species, such as, for example, Bremia lactucae; Peronospora species, such as, for example, Peronospora pisi or P. brassicae; Erysiphe species, such as, for example, Erysiphe graminis; Sphaerotheca species, such as, for example, Sphaerotheca fuliginea; Podosphaera species, such as, for example, Podosphaera leucotricha; Venturia species, such as, for example, Venturia inaequalis; Pyrenophora species, such as, for example, Pyrenophora teres or P. graminea (conidia form: Drechslera, syn: Helminthosporium); Cochliobolus species, such as, for example, Cochliobolus sativus (conidia form: Drechslera, syn: Helminthosporium); Uromyces species, such as, for example, Uromyces appendiculatus; Puccinia species, such as, for example, Puccinia recondita; Sclerotinia species, such as, for example, Sclerotinia sclerotiorum; Tilletia species, such as, for example, Tilletia caries; Ustilago species, such as, for example, Ustilago nuda or Ustilago avenae; Pellicularia species, such as, for example, Pellicularia sasakii; Pyricularia species, such as, for example, Pyricularia oryzae; Fusarium species, such as, for example, Fusarium culmorum; Botrytis species, such as, for example, Botrytis cinerea; Septoria species, such as, for example, Septoria nodorum; Leptosphaeria species, such as, for example, Leptosphaeria nodorum; Cercospora species, such as, for example, Cercospora canescens; Alternaria species, such as, for example, Alternaria brassicae; and Pseudocercosporella species, such as, for example, Pseudocercosporella herpotrichoides.\",\n \"The active compounds according to the invention also have very good fortifying action in plants.\",\n \"Accordingly, they can be used for mobilizing the defences of the plant against attack by undesirable microorganisms.\",\n \"In the present context, plant-fortifying (resistance-inducing) substances are to be understood as meaning those substances which are capable of stimulating the defence system of plants such that, when the treated plants are subsequently inoculated with undesirable microorganisms, they show substantial resistance against these microorganisms.\",\n \"In the present case, undesirable microorganisms are to be understood as meaning phytopathogenic fungi, bacteria and viruses.\",\n \"Accordingly, the substances according to the invention can be used to protect plants for a certain period after the treatment against attack by the pathogens mentioned.\",\n \"The period for which protection is provided generally extends over 1 to 10 days, preferably 1 to 7 days, after the treatment of the plants with the active compounds.\",\n \"The fact that the active compounds are well tolerated by plants at the concentrations required for controlling plant diseases permits the treatment of above-ground parts of plants, of propagation stock and seeds, and of the soil.\",\n \"The active compounds according to the invention are also suitable for increasing the yield of crops.\",\n \"In addition, they show reduced toxicity and are well tolerated by plants.\",\n \"At certain concentrations and application rates, the active compounds according to the invention can also be used as herbicides, for influencing plant growth and for controlling animal pests.\",\n \"If appropriate, they can also be used as intermediates and precursors for the synthesis of further active compounds.\",\n \"In the protection of materials, the compounds according to the invention can be employed for protecting industrial materials against infection with, and destruction by, undesired microorganisms.\",\n \"Industrial materials in the present context are understood as meaning non-living materials which have been prepared for use in industry.\",\n \"For example, industrial materials which are intended to be protected by active compounds according to the invention from microbial change or destruction can be adhesives, sizes, paper and board, textiles, leather, wood, paints and plastic articles, cooling lubricants and other materials which can be infected with, or destroyed by, microorganisms.\",\n \"Parts of production plants, for example cooling-water circuits, which may be impaired by the proliferation of microorganisms may also be mentioned within the scope of the materials to be protected.\",\n \"Industrial materials which may be mentioned within the scope of the present invention are preferably adhesives, sizes, paper and board, leather, wood, paints, cooling lubricants and heat-transfer liquids, particularly preferably wood.\",\n \"Microorganisms capable of degrading or changing the industrial materials which may be mentioned are, for example, bacteria, fungi, yeasts, algae and slime organisms.\",\n \"The active compounds according to the invention preferably act against fungi, in particular moulds, wood-discolouring and wood-destroying fungi (Basidiomycetes), and against slime organisms and algae.\",\n \"Microorganisms of the following genera may be mentioned as examples: Alternaria, such as Alternaria tenuis, Aspergillus, such as Aspergillus niger, Chaetomium, such as Chaetomium globosum, Coniophora, such as Coniophora puetana, Lentinus, such as Lentinus tigrinus, Penicillium, such as Penicillium glaucum, Polyporus, such as Polyporus versicolor, Aureobasidium, such as Aureobasidium pullulans, Sclerophoma, such as Sclerophoma pityophila, Trichoderma, such as Trichoderma viride, Escherichia, such as Escherichia coli, Pseudomonas, such as Pseudomonas aeruginosa, and Staphylococcus, such as Staphylococcus aureus.\",\n \"Depending on their particular physical and/or chemical properties, the active compounds can be converted into the customary formulations, such as solutions, emulsions, suspensions, powders, foams, pastes, granules, aerosols and microencapsulations in polymeric substances and in coating compositions for seeds, and ULV cool and warm fogging formulations.\",\n \"These formulations are produced in a known manner, for example by mixing the active compounds with extenders, that is, liquid solvents, liquefied gases under pressure, and/or solid carriers, optionally with the use of surfactants, that is emulsifiers and/or dispersants, and/or foam formers.\",\n \"If the extender used is water, it is also possible to employ, for example, organic solvents as auxiliary solvents.\",\n \"Essentially, suitable liquid solvents are: aromatics such as xylene, toluene or alkylnaphthalenes, chlorinated aromatics or chlorinated aliphatic hydrocarbons such as chlorobenzenes, chloroethylenes or methylene chloride, aliphatic hydrocarbons such as cyclohexane or paraffins, for example petroleum fractions, alcohols such as butanol or glycol and their ethers and esters, ketones such as acetone, methyl ethyl ketone, methyl isobutyl ketone or cyclohexanone, strongly polar solvents such as dimethylformamide and dimethyl sulphoxide, or else water.\",\n \"Liquefied gaseous extenders or carriers are to be understood as meaning liquids which are gaseous at standard temperature and under atmospheric pressure, for example aerosol propellants such as halogenated hydrocarbons, or else butane, propane, nitrogen and carbon dioxide.\",\n \"Suitable solid carriers are: for example ground natural minerals such as kaolins, clays, talc, chalk, quartz, attapulgite, montmorillonite or diatomaceous earth, and ground synthetic minerals such as finely divided silica, alumina and silicates.\",\n \"Suitable solid carriers for granules are: for example crushed and fractionated natural rocks such as calcite, marble, pumice, sepiolite and dolomite, or else synthetic granules of inorganic and organic meals, and granules of organic material such as sawdust, coconut shells, maize cobs and tobacco stalks.\",\n \"Suitable emulsifiers and/or foam formers are: for example nonionic and anionic emulsifiers, such as polyoxyethylene fatty acid esters, polyoxyethylene fatty alcohol ethers, for example alkylaryl polyglycol ethers, alkylsulphonates, alkyl sulphates, arylsulphonates, or else protein hydrolysates.\",\n \"Suitable dispersants are: for example lignosulphite waste liquors and methylcellulose.\",\n \"Tackifiers such as carboxymethylcellulose and natural and synthetic polymers in the form of powders, granules or latices, such as gum arabic, polyvinyl alcohol and polyvinyl acetate, or else natural phospholipids such as cephalins and lecithins and synthetic phospholipids can be used in the formulations.\",\n \"Other possible additives are mineral and vegetable oils.\",\n \"It is possible to use colorants such as inorganic pigments, for example iron oxide, titanium oxide and Prussian Blue, and organic dyestuffs such as alizarin dyestuffs, azo dyestuffs and metal phthalocyanine dyestuffs, and trace nutrients such as salts of iron, manganese, boron, copper, cobalt, molybdenum and zinc.\",\n \"The formulations generally comprise between 0.1 and 95% by weight of active compound, preferably between 0.5 and 90%.\",\n \"The active compounds according to the invention can be used as such or in their formulations, also in a mixture with known fungicides, bactericides, acaricides, nematicides or insecticides, to broaden, for example, the activity spectrum or to prevent development of resistance.\",\n \"In many cases, synergistic effects are obtained, i.e.\",\n \"the activity of the mixture is greater than the activity of the individual components.\",\n \"Examples of suitable mixing components are the following: Fungicides: aldimorph, ampropylfos, ampropylfos-potassium, andoprim, anilazine, azaconazole, azoxystrobin, benalaxyl, benodanil, benomyl, benzamacril, benzamacril-isobutyl, bialaphos, binapacryl, biphenyl, bitertanol, blasticidin-S, bromuconazole, bupirimate, buthiobate, calcium polysulphide, capsimycin, captafol, captan, carbendazim, carboxin, carvon, quinomethionate, chlobenthiazone, chlorfenazole, chloroneb, chloropicrin, chlorothalonil, chlozolinate, clozylacon, cufraneb, cymoxanil, cyproconazole, cyprodinil, cyprofuram, debacarb, dichlorophen, diclobutrazole, diclofluanid, diclomezine, dicloran, diethofencarb, difenoconazole, dimethirimol, dimethomorph, diniconazole, diniconazole-M, dinocap, diphenylamine, dipyrithione, ditalimfos, dithianon, dodemorph, dodine, drazoxolon, edifenphos, epoxiconazole, etaconazole, ethirimol, etridiazole, famoxadon, fenapanil, fenarimol, fenbuconazole, fenfuram, fenitropan, fenpiclonil, fenpropidin, fenpropimorph, fentin acetate, fentin hydroxide, ferbam, ferimzone, fluazinam, flumetover, fluoromide, fluquinconazole, flurprimidol, flusilazole, flusulfamide, flutolanil, flutriafol, folpet, fosetyl-alumninium, fosetyl-sodium, fthalide, fuberidazole, furalaxyl, furametpyr, furcarbonil, furconazole, furconazole-cis, furmecyclox, guazatine, hexachlorobenzene, hexaconazole, hymexazole, imazalil, imibenconazole, iminoctadine, iminoctadine albesilate, iminoctadine triacetate, iodocarb, ipconazole, iprobenfos (IBP), iprodione; irumamycin, isoprothiolane, isovaledione, kasugamycin, kresoxim-methyl, copper preparations, such as: copper hydroxide, copper naphthenate, copper oxychloride, copper sulphate, copper oxide, oxine-copper and Bordeaux mixture, mancopper, mancozeb, maneb, meferimzone, mepanipyrim, mepronil, metalaxyl, metconazole, methasulfocarb, methfuroxam, metiram, metomeclam, metsulfovax, mildiomycin, myclobutanil, myclozolin, nickel dimethyldithiocarbamate, nitrothal-isopropyl, nuarimol, ofurace, oxadixyl, oxamocarb, oxolinic acid, oxycarboxim, oxyfenthiin, paclobutrazole, pefurazoate, penconazole, pencycuron, phosdiphen, picoxystrobin, pimaricin, piperalin, polyoxin, polyoxorim, probenazole, prochloraz, procymidone, propamocarb, propanosine-sodium, propiconazole, propineb, pyraclostrobin, pyrazophos, pyrifenox, pyrimethanil, pyroquilon, pyroxyfur, quinconazole, quintozene (PCNB), sulphur and sulphur preparations, tebuconazole, tecloftalam, tecnazene, tetcyclacis, tetraconazole, thiabendazole, thicyofen, thifluzamide, thiophanate-methyl, thiram, tioxymid, tolclofos-methyl, tolylfluanid, triadimefon, triadimenol, triazbutil, triazoxide, trichlamide, tricyclazole, tridemorph, trifloxystrobin, triflumnizole, triforine, triticonazole, uniconazole, validamycin A, vinclozolin, viniconazole, zarilarnide, zineb, ziram and also Dagger G, OK-8705, OK-8801, α-(1,1-dimethylethyl)-β-(2-phenoxyethyl)-1H-1,2,4-triazole-1-ethanol, α-(2,4-dichlorophenyl)-β-fluoro-β-propyl-1H-1,2,4-triazole-1-ethanol, α-(2,4-dichlorophenyl)-β-methoxy-α-methyl-1H-1,2,4-triazole-1-ethanol, α-(5-methyl-1,3-dioxan-5-yl)-β-[[4-(trifluoromethyl)-phenyl]-methylene]-1H-1,2,4-triazole-1-ethanol, (5RS,6RS)-6-hydroxy-2,2,7,7-tetramethyl-5-(1H-1,2,4-triazol-1-yl)-3-octanone, (E)-α-(methoxyimino)-N-methyl-2-phenoxy-phenylacetamide, isopropyl {2-methyl-1-[[[1-(4-methylphenyl)-ethyl]-amino]-carbonyl]-propyl}-carbamate, 1-(2,4-dichlorophenyl)-2-(1H-1,2,4-triazol-1-yl)-ethanone O-(phenylmethyl)-oxime, 1-(2-methyl-1-naphthalenyl)-1H-pyrrole-2,5-dione, 1-(3,5-dichlorophenyl)-3-(2-propenyl)-2,5-pyrrolidinedione, 1-[(diiodomethyl)-sulphonyl]-4-methyl-benzene, 1-[[2-(2,4-dichlorophenyl)-1,3-dioxolan-2-yl]-methyl]-1H-imidazole, 1-[[2-(4-chlorophenyl)-3-phenyloxiranyl]-methyl]-1H-1,2,4-triazole, 1-[1-[2-[(2,4-dichlorophenyl)-methoxy]-phenyl]-ethenyl]-1H-imidazole, 1-methyl-5-nonyl-2-(phenylmethyl)-3-pyrrolidinole, 2′,6′-dibromo-2-methyl-4′-trifluoromethoxy-4′-trifluoro-methyl-1,3-thiazole-5-carboxanilide, 2,2-dichloro-N-[1-(4-chlorophenyl)-ethyl]-1-ethyl-3-methyl-cyclopropanecarboxamide, 2,6-dichloro-5-(methylthio)-4-pyrimidinyl-thiocyanate, 2,6-dichloro-N-(4-trifluoromethylbenzyl)-benzamide, 2,6-dichloro-N-[[4-(trifluoromethyl)-phenyl]-methyl]-benzamide, 2-(2,3,3-triiodo-2-propenyl)-2H-tetrazole, 2-[(1-methylethyl)-sulphonyl]-5-(trichloromethyl)-1,3,4-thiadiazole, 2-[[6-deoxy-4-O-(4-O-methyl-β-D-glycopyranosyl)-α-D-glucopyranosyl]-amino]-4-methoxy1H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile, 2-aminobutane, 2-bromo-2-(bromomethyl)-pentanedinitrile, 2-chloro-N-(2,3-dihydro-1,1,3-trimethyl-1H-inden-4-yl)-3-pyridinecarboxamide, 2-chloro-N-(2,6-dimethylphenyl)-N-(isothiocyanatomethyl)-acetamide, 2-phenylphenol (OPP), 3,4-dichloro-1-[4-(difluoromethoxy)-phenyl]-1H-pyrrole-2,5-dione, 3,5-dichloro-N-[cyano[(1-methyl-2-propynyl)-oxy]-methyl]-benzamide, 3-(1,1-dimethylpropyl)-1-oxo-1H-indene-2-carbonitrile, 3-[2-(4-chlorophenyl)-5-ethoxy-3-isoxazolidinyl]-pyridine, 4-chloro-2-cyano-N,N-dimethyl-5-(4-methylphenyl)-1H-imidazole-1-sulphonamide, 4-methyl-tetrazolo[1,5-a]quinazolin-5(4H)-one, 8-(1,1-dimethylethyl)-N-ethyl-N-propyl-1,4-dioxaspiro[4.5]decane-2-methaneamine, 8-hydroxyquinoline sulphate, 9H-xanthene-2-[(phenylamino)-carbonyl]-9-carboxylic hydrazide, bis-(1-methylethyl)-3-methyl-4-[(3-methylbenzoyl)-oxy]-2,5-thiophenedicarboxylate, cis-1-(4-chlorophenyl)-2-(1H-1,2,4-triazol-1-yl)-cycloheptanol, cis-4-[3-[4-(1,1-dimethylpropyl)-phenyl-2-methylpropyl]-2,6-dimethyl-morpholine-hydrochloride, ethyl [(4-chlorophenyl)-azo]-cyanoacetate, potassium hydrogen carbonate, methanetetrathiol sodium salt, methyl 1-(2,3-dihydro-2,2-dimethyl-1H-inden-1-yl)-1H-imidazole-5-carboxylate, methyl N-(2,6-dimethylphenyl)-N-(5-isoxazolylcarbonyl)-DL-alaninate, methyl N-(chloroacetyl)-N-(2,6-dimethylphenyl)-DL-alaninate, N-(2,3-dichloro-4-hydroxyphenyl)-1-methyl-cyclohexanecarboxamide, N-(2,6-dimethylphenyl)-2-methoxy-N-(tetrahydro-2-oxo-3-furanyl)-acetamide, N-(2,6-dimethylphenyl)-2-methoxy-N-(tetrahydro-2-oxo-3-thienyl)-acetamide, N-(2-chloro-4-nitrophenyl)-4-methyl-3-nitro-benzenesulphonamide, N-(4-cyclohexylphenyl)-1,4,5,6-tetrahydro-2-pyrimidineamine, N-(4-hexylphenyl)-1,4,5,6-tetrahydro-2-pyrimidineamine, N-(5-chloro-2-methylphenyl)-2-methoxy-N-(2-oxo-3-oxazolidinyl)-acetamide, N-(6-methoxy-3-pyridinyl)-cyclopropanecarboxamide, N-[2,2,2-trichloro-1-[(chloroacetyl)-amino]-ethyl]-benzamide, N-[3-chloro-4,5-bis-(2-propinyloxy)-phenyl]-N′-methoxy-methaneimidamide, N-formyl-N-hydroxy-DL-alanine-sodium salt, O,O-diethyl [2-(dipropylamino)-2-oxoethyl]-ethylphosphoramnidothioate, O-methyl S-phenyl phenylpropylphosphoramidothioate, S-methyl1,2,3-benzothiadiazole-7-carbothioate, spiro[2H]-1-benzopyrane-2,1′(3′H)-isobenzofuran]-3′-one, 4-[(3,4-dimethoxyphenyl)-3-(4-fluorophenyl)-acryloyl]-morpholine.\",\n \"Bactericides: bronopol, dichlorophen, nitrapyrin, nickel dimethyldithiocarbamate, kasugamycin, octhilinone, furancarboxylic acid, oxytetracyclin, probenazole, streptomycin, tecloftalam, copper sulphate and other copper preparations.\",\n \"Insecticides/acaricides/nematicides: abamectin, acephate, acetamiprid, acrinathrin, alanycarb, aldicarb, aldoxycarb, alpha-cypermethrin, alphamethrin, amitraz, avermectin, AZ 60541, azadirachtin, azamethiphos, azinphos A, azinphos M, azocyclotin, Bacillus popilliae, Bacillus sphaericus, Bacillus subtilis, Bacillus thuringiensis, baculoviruses, Beauveria bassiana, Beauveria tenella, bendiocarb, benfuracarb, bensultap, benzoximate, betacyfluthrin, bifenazate, bifenthrin, bioethanomethrin, biopermethrin, bistrifluron, BPMC, bromophos A, bufencarb, buprofezin, butathiofos, butocarboxim, butylpyridaben, cadusafos, carbaryl, carbofuran, carbophenothion, carbosulfan, cartap, chloethocarb, chlorethoxyfos, chlorfenapyr, chlorfenvinphos, chlorfluazuron, chlormephos, chlorpyrifos, chlorpyrifos M, chlovaporthrin, chromafenozide, cis-resmethrin, cispermnethrin, clocythrin, cloethocarb, clofentezine, clothianidine, cyanophos, cycloprene, cycloprothrin, cyfluthrin, cyhalothrin, cyhexatin, cypermethrin, cyromazine, deltamethrin, demeton M, demeton S, demeton-S-methyl, diafenthiuron, diazinon, dichlorvos, dicofol, diflubenzuron, dimethoat, dimethylvinphos, diofenolan, disulfoton, docusat-sodium, dofenapyn, eflusilanate, emamectin, empenthrin, endosulfan, Entomopfthora spp., esfenvalerate, ethiofencarb, ethion, ethoprophos, etofenprox, etoxazole, etrimfos; fenamiphos, fenazaquin, fenbutatin oxide, fenitrothion, fenothiocarb, fenoxacrim, fenoxycarb, fenpropathrin, fenpyrad, fenpyrithrin, fenpyroximate, fenvalerate, fipronil, fluazinam, fluazuron, flubrocythrinate, flucycloxuron, flucythrinate, flufenoxuron, flumethrin, flutenzine, fluvalinate, fonophos, fosmethilan, fosthiazate, fubfenprox, furathiocarb, granulosis viruses, halofenozide, HCH, heptenophos, hexaflumuron, hexythiazox, hydroprene, imidacloprid, indoxacarb, isazofos, isofenphos, isoxathion, ivermectin, nuclear polyhedrosis viruses, lambda-cyhalothrin, lufenuron, malathion, mecarbam, metaldehyde, methamidophos, Metharhizium anisopliae, Metharhizium flavoviride, methidathion, methiocarb, methoprene, methomyl, methoxyfenozide, metolcarb, metoxadiazone, mevinphos, milbemectin, milbemycin, monocrotophos, naled, nitenpyram, nithiazine, novaluron, omethoate, oxamyl, oxydemethon M, Paecilomyces fumosoroseus, parathion A, parathion M, permethrin, phenthoate, phorat, phosalone, phosmet, phosphamidon, phoxim, pirimicarb, pirimiphos A, pirimiphos M, profenofos, promecarb, propargite, propoxur, prothiofos, prothoat, pymetrozine, pyraclofos, pyresmethrin, pyrethrum, pyridaben, pyridathion, pyrimidifen, pyriproxyfen, quinalphos, ribavirin, salithion, sebufos, silafluofen, spinosad, spirodiclofen, sulfotep, sulprofos, tau-fluvalinate, tebufenozide, tebufenpyrad, tebupirimiphos, teflubenzuron, tefluthrin, temephos, temivinphos, terbufos, tetrachlonrinphos, tetradifon theta-cypermethrin, thiacloprid, thiamethoxam, thiapronil, thiatriphos, thiocyclam hydrogen oxalate, thiodicarb, thiofanox, thuringiensin, tralocythrin, tralomethrin, triarathene, triazamate, triazophos, triazuron, trichlophenidine, trichlorfon, triflumuron, trimethacarb, vamidothion, vaniliprole, Verticillium lecanii, YI5302 zeta-cypermethfin, zolaprofos (1R-cis)-[5-(phenylmethyl)-3-furanyl]-methyl-3-[(dihydro-2-oxo-3(2H)-furanylidene)-methyl]-2,2-dimethylcyclopropanecarboxylate, (3-phenoxyphenyl)-methyl-2,2,3,3-tetramethylcyclopropanecarboxylate, 1-[(2-chloro-5-thiazolyl)methyl]tetrahydro-3,5-dimethyl-N-nitro-1,3,5-triazine-2(1H)-imine, 2-(2-chloro-6-fluorophenyl)-4-[4-(1,1-dimethylethyl)phenyl]-4,5-dihydro-oxazole, 2-(acetyloxy)-3-dodecyl-1,4-naphthalenedione, 2-chloro-N-[[[4-(1-phenylethoxy)-phenyl]-amino]-carbonyl]-benzamide, 2-chloro-N-[[[4-(2,2-dichloro-1,1-difluoroethoxy)-phenyl]-amino]-carbonyl]-benzamide, 3-methylphenyl propylcarbamate, 4-[4-(4-ethoxyphenyl)-4-methylpentyl]-1-fluoro-2-phenoxy-benzene, 4-chloro-2-(1,1-dimethylethyl)-5-[[2-(2,6-dimethyl-4-phenoxyphenoxy)ethyl]thio]-3(2H)-pyridazinone, 4-chloro-2-(2-chloro-2-methylpropyl)-5-[(6-iodo-3-pyridinyl)methoxy]-3(2H)-pyridazinone, 4-chloro-5-[(6-chloro-3-pyridinyl)methoxy]-2-(3,4-dichlorophenyl)-3(2H)-pyridazinone, Bacillus thuringiensis strain EG-2348, [2-benzoyl-1-(1,1-dimethylethyl)-hydrazinobenzoic acid, 2,2-dimethyl-3-(2,4-dichlorophenyl)-2-oxo-1-oxaspiro[4.5]dec-3-en4-yl butanoate, [3-[(6-chloro-3-pyridinyl)methyl]-2-thiazolidinylidene]-cyanamide, dihydro-2-(nitromethylene)-2H-1,3-thiazine-3(4H)-carboxaldehyde, ethyl [2-[[1,6-dihydro-6-oxo-1-(phenylmethyl)-4-pyridazinyl]oxy]ethyl]-carbamate, N-(3,4,4-trifluoro-1-oxo-3-butenyl)-glycine, N-(4-chlorophenyl)-3-[4-(difluoromethoxy)phenyl]-4,5-dihydro4-phenyl-1H-pyrazole-1-carboxamide, N-[(2-chloro-5-thiazolyl)methyl]-N′-methyl-N″-nitro-guanidine, N-methyl-N′-(1-methyl-2-propenyl)-1,2-hydrazinedicarbothioamide, N-methyl-N′-2-propenyl-1,2-hydrazinedicarbothioamide, O,O-diethyl [2-(dipropylamino)-2-oxoethyl]-ethylphosphoramidothioate, N-cyanomethyl-4-trifluoromethyl-nicotinamide, 3,5-dichloro-1-(3,3-dichloro-2-propenyloxy)-4-[3-(5-trifluoromethylpyridine-2-yloxy)-propoxy]-benzene.\",\n \"A mixture with other known active compounds, such as herbicides, or with fertilizers and growth regulators, is also possible.\",\n \"In addition, the compounds of the formula (I) according to the invention also have very good antimycotic activity.\",\n \"They have a very broad antimycotic activity spectrum in particular against dermatophytes and yeasts, moulds and diphasic fungi (for example against Candida species, such as Candida albicans, Candida glabrata), and Epidermophyton floccosum, Aspergillus species, such as Aspergillus niger and Aspergillus fumigatus, Trichophyton species, such as Trichophyton mentagrophytes, Microsporon species such as Microsporon canis and audouinii.\",\n \"The list of these fungi by no means limits the mycotic spectrum covered, but is only for illustration.\",\n \"The active compounds can be used as such, in the form of their formulations or the use forms prepared therefrom, such as ready-to-use solutions, suspensions, wettable powders, pastes, soluble powders, dusts and granules.\",\n \"Application is carried out in a customary manner, for example by watering, spraying, atomizing, broadcasting, dusting, foaming, spreading, etc.\",\n \"It is furthermore possible to apply the active compounds by the ultra-low-volume method, or to inject the active compound preparation or the active compound itself into the soil.\",\n \"It is also possible to treat the seeds of the plants.\",\n \"When using the active compounds according to the invention as fungicides, the application rates can be varied within a relatively wide range, depending on the kind of application.\",\n \"For the treatment of parts of plants, the active compound application rates are generally between 0.1 and 10,000 g/ha, preferably between 10 and 1000 g/ha.\",\n \"For seed dressing, the active compound application rates are generally between 0.001 and 50 g per kilogram of seed, preferably between 0.01 and 10 g per kilogram of seed.\",\n \"For the treatment of the soil, the active compound application rates are generally between 0.1 and 10,000 g/ha, preferably between 1 and 5000 g/ha.\",\n \"The preparation and the use of the active compounds according to the invention is illustrated by the examples below.\",\n \"PREPARATION EXAMPLES Example I-1-a-1 1.65 g (14.4 mmol) of potassium tert-butoxide are initially charged in 30 ml of anhydrous dimethylformamide (DMF), and 3.5 g (9.6 mmol) of the compound according to Example (II-1) in 10 ml of anhydrous DMF are added dropwise.\",\n \"The mixture is stirred at 50° C. for 3 h. 100 ml of ice-water are then poured into the reaction solution.\",\n \"This mixture is poured into 600 ml of cold 1N HCl solution.\",\n \"The precipitate is filtered off with suction and dried.\",\n \"Yield: 2.9 g (90% of theory), m.p.\",\n \"139-141° C. The following compounds of the formula (I-1-a) were prepared analogously to Example (I-1-a-1) and in accordance with the general statements about the preparation: (I-1-a) Ex.\",\n \"No.\",\n \"X Y B A Q1 Q2 m.p.° C. I-1-a-2 CH3 4-Cl—C6H4 —(CH2)5— H H 105-107° C. I-1-a-3 C2H5 4-Cl—C6H4 —(CH2)5— H H 121-123° C. I-1-a-4 CH3 4-Cl—C6H4 —(CH2)2—O—(CH2)4— H H 130 I-1-a-5 CH3 4-CF3—C6H4 —(CH2)5— H H Oil I-1-a-6 CH3 4-Cl—C6H4 H —(CH2)4— H 145-148 I-1-a-7 CH3 4-CH3—C6H4 —(CH2)5— H H 85 I-1-a-8 CH3 4-Cl—C6H4—O—C6H4 —(CH2)5— H H 75-77 I-1-a-9 CH3 4-Cl—C6H4 —(CH2)2—CHCH3—(CH2)2— H H 142-144 I-1-a-10 CH3 4-Cl—C6H4 —(CH2)2—CHOCH3—(CH2)2— H H 127 I-1-a-11 CH3 4-Cl—C6H4 H C6H5— H H 130 I-1-a-12 C2H5 4-Cl—C6H4 —(CH2)2—O—(CH2)2— H H 210 I-1-a-13 CH3 4-Cl—C6H4 H C6H11— H H 105 I-1-a-14 CH3 4-Cl—C6H4 —(CH2)2—CH—C3H7—(CH2)2— H H oil I-1-a-15 CH3 4-Cl—C6H4 —(CH2)2—CH-t-C4H9—(CH2)2— H H oil I-1-a-16 CH3 4-Cl—C6H4 —(CH2)2—CH—C2H5—(CH2)2— H H oil I-1-a-17 H 4-Cl—C6H4 —(CH2)5— H H 185 Example I-1-b-1 500 mg (1.33 mmol) of the compound according to Example I-1-a-2 are initially charged in 5 ml of absolute dichloromethane, and 0.27 ml (2.0 mmol) of triethylamine is added.\",\n \"At 0° C., 0.30 g (1.74 mmol) of 6-chloronicotinyl chloride is added, and the mixture is stirred at room temperature for 2 h. A further 0.15 g of chloronicotinyl chloride and 0.13 ml of triethylamine are then added.\",\n \"The reaction mixture is washed with 10% citric acid solution, the aqueous phase is extracted with dichloromethane, and the organic phase is then washed with 1N aqueous sodium hydroxide solution and the aqueous phase is extracted with dichloromethane.\",\n \"The organic phase is dried and the solvent is distilled off.\",\n \"The residue is stirred with petroleum ether and filtered and then dried.\",\n \"Yield: 0.7 g (100% of theory), m.p.\",\n \"133-134° C. The following compounds of the formula (I-1-b) were obtained analogously to Example (1-1-b-1) and in accordance with the general statements about the preparation: (I-1-b) Ex.\",\n \"No.\",\n \"X Y B A Q1 Q2 R1 m.p.° C. I-1-b-2 CH3 4-Cl—C6H4 —(CH2)5— H H H5C2—O—CH2— 135 I-1-b-3 CH3 4-Cl—C6H4 —(CH2)5— H H i-C3H7 oil I-1-b-4 CH3 4-Cl—C6H4 —(CH2)5— H H 4-Cl—C6H4 129-130 I-1-b-5 CH3 4-Cl—C6H4 —(CH2)5— H H Oil Example I-1-c-1 500 mg (1.5 mmol) of the compound according to Example (I-1-a-1) are initially charged in 5 ml of absolute dichloromethane, and 0.3 ml (2.25 mmol) of triethylamine is added.\",\n \"At 0° C., 0.225 g (1.95 mmol) of ethyl chloroformate is added, and the mixture is stirred at room temperature for 2 h. The reaction solution is washed with 10% strength citric acid and the aqueous phase is extracted with dichloromethane (CH2Cl2).\",\n \"The organic phase is washed with 1N sodium hydroxide solution, dried and concentrated.\",\n \"Yield: 0.45 g (75% of theory), 132-134° C. The following compounds of the formula (I-1-c) were obtained analogously to Example (I-1-c-1) and in accordance with the general statements about the preparation: (I-1-c) Ex.No.\",\n \"X Y A B Q1 Q2 L M R2 m.p.° C. I-1-c-2 CH3 4-Cl—C6H4 —(CH2)5— H H O O C25 oil I-1-c-3 C2H5 4-Cl—C6H4 —(CH2)5— H H O O C25 oil I-1-c-4 CH3 4-Cl—C6H4 —(CH2)5— H H O O C65 139-140 I-1-c-5 CH3 4-Cl—C6H4 —(CH2)5— H H O S C65—CH2 144 I-1-c-6 CH3 4-Cl—C6H4 —(CH2)5— H H O O C65—CH2 137 Example I-1-d-1 0.5 g (1.33 mmol) of the compound according to Example I-1-a-2 is initially charged in 5 ml of anhydrous dichloromethane, and 0.27 ml (2.0 mmol) of triethylamine is added.\",\n \"With ice-cooling, 0.135 ml (1.74 mmol) of methanesulphonyl chloride is added.\",\n \"The mixture is stirred at room temperature for 2 h. The reaction mixture is extracted with 10% strength citric acid, the aqueous phase is washed with dichloromethane, the organic phase is extracted with 1N sodium hydroxide solution and the aqueous phase is washed with dichloromethane, dried and concentrated.\",\n \"Yield: 0.6 g (100% of theory), m.p.\",\n \"134° C. 1H-NMR (d6-DMSO, 400 MHz): δ=1.02-1.75 (m, 10H, cyc-hexyl-CH2), 2.30 (s, 3H, thiazolyl-CH3), 3.05 (s, 2H, CH2—CO), 3.61 (s, 3H, SO2—CH3), 7.54 (d, 2H, Ar-H), 7.86 (d, 2H, Ar-H) ppm.\",\n \"Example I-1-g-1 0.5 g (1.33 mmol) of the compound according to Example I-1-a-2 and 0.27 ml (2.0 mmol) of triethylamine are dissolved in 5 ml of anhydrous dichloromethane, and 0.26 g (1.73 mmol) of morpholine N-carbonyl chloride are added with ice-cooling.\",\n \"The mixture is stirred at room temperature overnight.\",\n \"The reaction mixture is extracted with 10% strength citric acid, the aqueous phase is washed with dichloromethane, the organic phase is extracted with 1N sodium hydroxide solution and the aqueous phase is washed with dichloromethane, dried and concentrated.\",\n \"Yield: 0.69 g (93% of theory), wax 1H-NMR (d6-DMSO, 400 MHz): δ=1.02-1.74 (m, 10H, cyc-hexyl-CH2), 2.30 (s, 3H, thiazolyl-CH3), 3.01 (s, 2H, CH2—CO), 3.05-3.79 (m, 8H, morpholine-CH2), 7.54 (d, 2H, Ar-H), 7.85 (d, 2H, Ar-H) ppm.\",\n \"Example II-1 17.3 g (30 mmol) of the compound according to Example XIV-1 are initially charged in 200 ml of anhydrous acetone, and 6.8 g (49 mmol) of potassium carbonate and 20.5 g (147 mmol) of methyl iodide are added.\",\n \"The mixture is stirred under reflux for 16 h. The solution is filtered, the solvent is distilled off and the residue is purified by silica gel chromatography (dichloromethane:petroleum ether, 4:1→dichloromethane→dichloromethane:ethyl acetate, 30:1→10:1).\",\n \"Yield: 3.5 g (31% of theory), oil 1H-NMR (d6-DMSO, 400 MHz): δ=1.11 (s, 6H, 2-CH3-C-aliph.\",\n \"), 2.31 (s, 3H, CH3-C-heteroarom.)\",\n \"3.50 (s, 3H, CH3—O), 7.50 (d, 2H, 2-CH-arom.\",\n \"), 7.84 (d, 2H, 2-CH-arom) ppm.\",\n \"Example XIV-1 A solution of 12.7 g (45 mmol) of methyl 4-[2-(4-chlorophenyl)-5-methyl]-thiazolyl-acetate in 15 ml of tetrahydrofuran is, at −15° C., added dropwise to a solution of 25 ml of LDA* solution (2 molar) in 50 ml of anhydrous tetrahydrofuran, and the mixture is stirred at 0° C. for 60 min.\",\n \"At −15° C., a solution of 3.8 g (30 mmol) of 2,2-dimethylsuccinic anhydride in 10 ml of anhydrous tetrahydrofuran is then added dropwise.\",\n \"The solution is stirred at room temperature for two hours, 75 ml of water and 20 g of ammonium chloride are then added and the solution is acidified using concentrated hydrochloric acid.\",\n \"The intermediate is extracted with ether and the solvent is distilled off.\",\n \"The residue is boiled under reflux with 25 g KOH and 170 ml of water for one day.\",\n \"The mixture is cooled, acidified with conc.\",\n \"hydrochloric acid and extracted with ether.\",\n \"The crude product is directly, without further characterization and purification, converted into the compound II-1.Yield: 17.5 g (100% of theory), oil *LDA =lithium diisopropylamide The following compounds of the formula (II) were obtained analogously to Example (II-1) and in accordance with the general statements about the preparation: (II) Ex.\",\n \"No.\",\n \"X Y B A Q1 Q2 R8 m.p.° C. II-2 CH3 4-Cl—C6H4 —(CH2)5— H H CH3 oil II-3 C2H5 4-Cl—C6H4 —(CH2)5— H H CH3 oil II-4 CH3 4-Cl—C6H4 —(CH2)2—O—(CH2)2— H H CH3 oil II-5 CH3 4-CF3—C6H4 —(CH2)5— H H CH3 oil II-6 CH3 4-Cl—C6H4 H —(CH2)4— H CH3 oil II-7 CH3 4-CH3—C6H4 —(CH2)5— H H CH3 oil II-8 CH3 4-Cl—C6H4—O—C6H4 —(CH2)5— H H CH3 oil II-9 CH3 4-Cl—C6H4 —(CH2)2—CHCH3—(CH2)2— H H CH3 oil II-10 CH3 4-Cl—C6H4 —(CH2)2—CHOCH3—(CH2)2— H H CH3 oil II-11 CH3 4-Cl—C6H4 H C6H5— H H CH3 oil II-12 C2H5 4-Cl—C6H4 —(CH2)2—O—(CH2)2— H H CH3 oil II-13 CH3 4-Cl—C6H4 H C6H11 H H CH3 oil II-14 CH3 4-Cl—C6H4 —(CH2)2—CH—C3H7—(CH2)2— H H CH3 oil II-15 CH3 4-Cl—C6H4 —(CH2)2—CH-t-C4H9—(CH2)2— H H CH3 oil II-16 CH3 4-Cl—C6H4 —(CH2)2—CH—C2H5—(CH2)2— H H CH3 oil II-17 CH3 4-Cl—C6H4 —(CH2)5— H H CH3 oil The following compounds of the formula (XIV) were obtained analogously to Example (XIV-1) and in accordance with the general statements about the preparation: (XIV) Ex.\",\n \"No.\",\n \"X Y B A Q1 Q2 m.p.° C. XIV-2 CH3 4-Cl—C6H4 —(CH2)5— H H wax XIV-3 C2H5 4-Cl—C6H4 —(CH2)5— H H wax XIV-4 CH3 4-Cl—C6H4 —(CH2)2—O—(CH2)2— H H oil XIV-5 CH3 4-CF3—C6H4 —(CH2)5— H H oil XIV-6 CH3 4-Cl—C6H4 H —(CH2)4— H oil XIV-7 CH3 4-CH3—C6H4 —(CH2)5— H H oil XIV-8 CH3 4-Cl—C6H4—O—C6H4 —(CH2)5— H H oil XIV-9 CH3 4-Cl—C6H4 —(CH2)2—CHCH3—(CH2)2— H H oil XIV-10 CH3 4-Cl—C6H4 —(CH2)2—CHOCH3—(CH2)2— H H oil XIV-11 CH3 4-Cl—C6H4 H C6H5— H H oil XIV-12 C2H5 4-Cl—C6H4 —(CH2)2—O—(CH2)2— H H oil XIV-13 CH3 4-Cl—C6H4 H C6H11— H H oil XIV-14 CH3 4-Cl—C6H4 —(CH2)2—CH—C3H7—(CH2)2— H H oil XIV-15 CH3 4-Cl—C6H4 —(CH2)2—CH-t-C4H9—(CH2)2— H H oil XIV-16 CH3 4-Cl—C6H4 —(CH2)2—CH—C2H5—(CH2)2— H H oil XIV-17 H 4-Cl—C6H4 —(CH2)5— H H oil Example 1-2-a-1 2.16 g (18.1 mmol) of potassium tert-butoxide are initially charged in 20 ml of anhydrous dimethylformamide, and 4.6 g (12.1 mmol) of the compound according to Example III-1 in 2 ml of anhydrous dimethylformamide are added dropwise.\",\n \"The mixture is stirred at 50° C. for 2 hours.\",\n \"The reaction solution is poured into 100 ml of ice-water, and this mixture is stirred into 500 ml of cold 1N hydrochloric acid solution.\",\n \"The precipitate is filtered off with suction and taken up in dichloromethane.\",\n \"The solvent is distilled off, and the residue is then purified by silica gel chromatography (cyclohexane/ethyl acetate, 5:1).\",\n \"Yield: 1.4 g (13% of theory), m.p.\",\n \"165-167° C. The following compounds of the formula (I-2-a) were obtained analogously to Example (I-2-a-1) and in accordance with the general statements about the preparation: (I-2-a) Ex.\",\n \"No.\",\n \"X Y A B Q1 Q2 Q3 Q4 m.p.° C. I-2-a-2 CH3 4-Cl—C6H4 H H H H H H 106 I-2-a-3 CH3 4-Cl—C6H4 CH3 CH3 H H H H oil I-2-a-4 CH3 4-Cl—C6H4 H H —(CH2)4— H H 122 I-2-a-5 CH3 4-Cl—C6H4 H H 4-Cl—C6H4 H H H 195-197 Example 1-2-b-1 500 mg (1.44 mmol) of the compound according to Example I-2-a-1 are initially charged in 5 ml of anhydrous dichloromethane, and 0.29 ml (2.16 mmol) of triethylamine is added.\",\n \"At 0° C., 0.33 g (1.87 mmol) of 6-chloronicotinyl chloride is added, and the mixture is stirred at room temperature for 2 h. The reaction mixture is extracted with 10% citric acid, the aqueous phase is washed with dichloromethane, the organic phase is extracted with 1N sodium hydroxide solution and the aqueous phase is washed with dichloromethane, dried and concentrated.\",\n \"Yield: 0.7 g (100% of theory), m.p.\",\n \"127-130° C. The following compounds of the formula (I-2-b) were obtained analogously to Example (I-2-b-1) and in accordance with the general statements about the preparation: (I-2-b) Ex.\",\n \"No.\",\n \"X Y A B Q1 Q2 Q3 Q4 R1 m.p.° C. I-2-b-2 CH3 4-Cl—C6H4 H H CH3 CH3 H H i-C3H7— 96 I-2-b-3 CH3 4-Cl—C6H4 H H CH3 CH3 H H H5C2—O—CH2— 142 I-2-b-4 CH3 4-Cl—C6H4 H H CH3 CH3 H H 4-Cl—C6H4 123-125 I-2-b-5 CH3 4-Cl—C6H4 H H CH3 CH3 H H 125 Example 1-2-c-1 600 mg (1.7 mmol) of the compound according to Example I-2-a-1 are initially charged in 5 ml of anhydrous dichloromethane, and 0.35 ml (2.60 mmol) of triethylamine is added.\",\n \"At 0° C., 0.255 g (2.21 mmol) of ethyl chloroformate is added, and the mixture is stirred at room temperature for 2 hours.\",\n \"The solution is washed with 10% strength citric acid and the aqueous phase is extracted with dichloromethane.\",\n \"The organic phase is washed with 1N sodium hydroxide solution, dried and concentrated.\",\n \"Yield: 0.62 g (87% of theory), m.p.\",\n \"82-86° C. The following compounds of the formula (I-2-c) were obtained analogously to Example (I-2-c-1) and in accordance with the general statements about the preparation: (I-2-c) Ex.\",\n \"No.\",\n \"X Y A B Q1 Q2 Q3 Q4 M R2 m.p.° C. I-2-c-2 CH3 4-Cl—C6H4 H H CH3 CH3 H H O C6H5— 123-125 I-2-c-3 CH3 4-Cl—C6H4 H H CH3 CH3 H H O C6H5—CH2 129-131 I-2-c-4 CH3 4-Cl—C6H4 H H CH3 CH3 H H S C6H5—CH2 94 Example No.\",\n \"I-2-d-1 500 mg (1.44 mmol) of the compound according to Example I-2-a-1 are initially charged in 5 ml of anhydrous dichloromethane, and 0.29 ml (2.16 mmol) of triethylamine is added.\",\n \"At 0° C., 0.22 g (1.87 mmol) of methylsulphonyl chloride is added, and the mixture is stirred at room temperature for 2 h. The reaction mixture is extracted with 10% strength citric acid, the aqueous phase is washed with dichloromethane, the organic phase is extracted with 1N NaOH and the aqueous phase is washed with dichloromethane, dried and concentrated.\",\n \"Yield: 0.65 g (100% of theory), wax 1H-NMR (d6-DMSO, 400 MHz): δ=1.17 (s, 6H, cyc-hexyl-CH3), 2.25 (s, 3H, thiazolyl-CH3), 2.48 (s, 2H, cyc-hexyl-CH2), 2.86 (s, 2H, cyc-hexyl-CH2), 3.21 (s, 3H, SO2—CH3), 7.52 (d, 2H, Ar-H) 7.83 (d, 2H, Ar-H) ppm.\",\n \"Example No.\",\n \"I-2-g-1 500 mg (1.44 mmol) of the compound according to Example I-2-a-1 are initially charged in 5 ml of anhydrous dichloromethane, and 0.29 ml (2.16 mmol) of triethylamine is added.\",\n \"At 0° C., 0.27 g (1.87 mmol) of morpholinecarbonyl chloride is added, and the mixture is stirred at room temperature for 2 h. The reaction mixture is extracted with 10% strength citric acid, the aqueous phase is washed with dichloromethane, the organic phase is extracted with 1N sodium hydroxide solution and the aqueous phase is washed with dichloromethane, dried and concentrated.\",\n \"Yield: 0.65 g (100% of theory), m.p.\",\n \"128° C. 1H-NMR (d6-DMSO, 400 MHz): δ=1.13 (s, 6H, cyc-hexyl-CH3), 2.21 (s, 3H, thiazolyl-CH3), 2.46 (s, 2H, cyc-hexyl-CH2), 2.71 (s, 2H, cyc-hexyl-CH2), 3.06-3.80 (m, 8H, morpholine-CH2), 7.53 (d, 2H, Ar-H), 7.84 (d, 2H, Ar-H) ppm.\",\n \"Example III-1 17.0 g (30 mmol) of crude product of the compound according to Example XIX-1 are initially charged in 200 ml of anhydrous acetone, and 6.42 g (46.5 mmol) of potassium carbonate and 19.5 g (139.5 mmol) of methyl iodide are added.\",\n \"The mixture is stirred under reflux for 16 h. The mixture is filtered and concentrated.\",\n \"The residue is purified by silica gel chromatography (dichloromethane:petroleum ether, 4:1→dichloromethane→dichloromethane:ethyl acetate, 30:1→10:1) Yield: 4.6 g (16% of theory), oil 1H-NMR (400 MHz, d6-DMSO): δ=1.03 (s, 6H, 2—CH3-C-aliph.\",\n \"), 2.32 (s, 3H, CH3—C-arom.)\",\n \"7.50 (d, 2H, Ar-H.), 7.81 (d, 2H, Ar-H) ppm.\",\n \"Example XIX-1 A solution of 12.7 g (45 mmol) of methyl 4-[2-(4-chlorophenyl)-5-methyl]-thiazolyl-acetate in 15 ml of anhydrous tetrahydrofuran is, at −15° C., added dropwise to a solution of 25 ml of LDA solution (2 molar) in 50 ml of anhydrous tetrahydrofuran, and the mixture is stirred at 0° C. for 60 min.\",\n \"At −15° C., a solution of 4.26 g (30 mmol) of 3,3-dimethylglutaric anhydride in 10 ml of tetrahydrofuran is then added dropwise.\",\n \"The solution is stirred at room temperature for 2 hours, 75 ml of water and 20 g of ammonium chloride are then added and the mixture is then acidified using concentrated hydrochloric acid.\",\n \"The intermediate is extracted with ether and the solvent is distilled off.\",\n \"The residue is boiled under reflux with 25 g of potassium hydroxide and 170 ml of water for one day.\",\n \"The mixture is cooled, acidified with concentrated hydrochloric acid and extracted with ether.\",\n \"The crude product is directly, without purification, reacted further.\",\n \"Yield: 17 g (100% of theory), oil The following compounds of the formula (III) were obtained analogously to Example (III-1) and in accordance with the general statements about the preparation: (III) Ex.\",\n \"No.\",\n \"X Y A B Q1 Q2 Q3 Q4 R8 m.p.° C. III-2 CH3 4-Cl—C6H4 H H H H H H CH3 oil III-3 CH3 4-Cl—C6H4 CH3 CH3 H H H H CH3 oil III-4 CH3 4-Cl—C6H4 H H —(CH2)4— H H CH3 oil III-5 CH3 4-Cl—C6H4 H H 4-Cl—C6H4 H H H CH3 oil The following compounds of the formula (XIX) were obtained analogously to Example (XIX-1) and in accordance with the general statements about the preparation: (XIX) Ex.\",\n \"No.\",\n \"X Y A B Q1 Q2 Q3 Q4 m.p.° C. XIX-2 CH3 4-Cl—C6H4 H H H H H H oil XIX-3 CH3 4-Cl—C6H4 CH3 CH3 H H H H oil XIX-4 CH3 4-Cl—C6H4 H H —(CH2)4— H H oil XIX-5 CH3 4-Cl—C6H4 H H 4-Cl—C6H4 H H H oil Use Examples Example A Meloidogyne Test Solvent: 30 parts by weight of dimethylformamide Emulsifier: 1 part by weight of alkylaryl polyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amounts of solvent and emulsifier, and the concentrate is diluted with water to the desired concentration.\",\n \"Containers are filled with sand, solution of active compound, Meloidogyne incognita egg/larvae suspension and lettuce seeds.\",\n \"The lettuce seeds germinate the plants develop.\",\n \"On the roots, galls are formed.\",\n \"After the desired period of time, the nematicidal action is determined in % by the gall formation.\",\n \"100% means that no galls have been found; 0% means that the number of galls on the treated plants corresponds to that of the untreated control.\",\n \"In this test, for example, the following compounds of the preparation examples showed good activity: Concentration of active Active compounds compound in ppm Kill rate in % after 14 d Ex.\",\n \"I-1-c-3 20 100 Ex.\",\n \"I-1-a-1 20 95 Example B Myzus Test (Systemic Action) Solvent: 7 parts by weight of dimethylformamide Emulsifier: 2 parts by weight of alkylaryl polyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amounts of solvent and emulsifier, and the concentrate is diluted with emulsifier-containing water to the desired concentration.\",\n \"Savoy cabbage (Brassica oleracea) which is heavily infested by the green peach aphid (Myzus persicae) is watered with in each case 10 ml of preparation of active compound of the desired concentration such that the preparation of active compound penetrates into the soil without wetting the seedling.\",\n \"The active compound is taken up by the roots and transferred into the seedling.\",\n \"After the desired period of time, the kill in % is determined.\",\n \"100% means that all aphids have been killed; 0% means that none of the aphids have been killed.\",\n \"In this test, for example, the following compounds of the preparation examples show good activity: Concentration of active Active compounds compound in ppm Kill rate in % after 10 d Ex.\",\n \"I-1-c-2 20 95 Example C Phaedon Larvae Test Solvent: 7 parts by weight of dimethylformamide Emulsifier: 2 parts by weight of alkylaryl polyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amounts of solvent and emulsifier, and the concentrate is diluted with emulsifier-containing water to the desired concentration.\",\n \"Cabbage leaves (Brassica oleracea) are treated by being dipped into the preparation of active compound of the desired concentration and are populated with larvae of the mustard beetle (Phaedon cochleariae) while the leaves are still moist.\",\n \"After the desired period of time, the kill in % is determined.\",\n \"100% means that all beetle larvae have been killed; 0% means that none of the beetle larvae have been killed.\",\n \"In this test, for example, the following compounds of the preparation examples show good activity: Concentration of active Active compounds compound in ppm Kill rate in % after 7 d Ex.\",\n \"I-1-c-1 500 100 Ex.\",\n \"I-2-c-1 500 100 Example D Spodoptera Frugiperda Test Solvent: 7 parts by weight of dimethylfornamide Emulsifier: 2 parts by weight of alkylaryl polyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amounts of solvent and emulsifier, and the concentrate is diluted with emulsifier-containing water to the desired concentration.\",\n \"Cabbage leaves (Brassica oleracea) are treated by being dipped into the preparation of active compound of the desired concentration and are populated with caterpillars of the armyworm (Spodoptera frugiperda) while the leaves are still moist.\",\n \"After the desired period of time, the kill in % is determined.\",\n \"100% means that all caterpillar larvae have been killed; 0% means that none of the caterpillar have been killed.\",\n \"In this test, for example, the following compounds of the preparation examples show good activity: Concentration of active Active compound compound in ppm Kill rate in % after 7 d Ex.\",\n \"I-1-a-2 500 100 Ex.\",\n \"I-2-a-1 500 100 Ex.\",\n \"I-1-c-1 500 100 Ex.\",\n \"I-2-c-1 500 100 Example E Tetranychus test (OP-Resistant/Dip Treatment) Solvent: 30 parts by weight of dimethylformamide Emulsifier: 1 part by weight of alkylaryl polyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amounts of solvent and emulsifier, and the concentrate is diluted with emulsifier-containing water to the desired concentration.\",\n \"Bean plants (Phaseolus vulgaris) which are heavily infested by all stages of the greenhouse red spider mite (Tetranychus urticae) are dipped into a preparation of active compound of the desired concentration.\",\n \"After the desired period of time, the effect in % is determined.\",\n \"100% means that all spider mites have been killed; 0% means that none of the spider mites have been killed.\",\n \"In this test, for example, the following compounds of the preparation examples show good activity: Concentration of active Active compound compound in ppm Kill rate in % after 7 d Ex.\",\n \"I-1-c-3 100 98 Ex.\",\n \"I-1-a-1 100 100 Ex.\",\n \"I-2-a-2 100 99 Ex.\",\n \"I-2-a-1 100 98 Ex.\",\n \"I-1-c-1 100 95 Ex.\",\n \"I-2-c-1 100 100 Example F1 Post-Emergence Test Solvent: 5 parts by weight of acetone Emulsifier: 1 part by weight of alkylaryl polyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amount of solvent, the stated amount of emulsifier is added and the concentrate is diluted with water to the desired concentration.\",\n \"Test plants of a height of 5-15 cm are sprayed with the preparation of active compound such that the particular amounts of active compound desired are applied per unit area.\",\n \"The concentration of the spray liquor is chosen such that the particular amounts of active compound desired are applied in 1000 l of water/ha.\",\n \"After three weeks, the degree of damage to the plants is rated in % damage in comparison to the development of the untreated control.\",\n \"The figures denote: 0% =no effect (like untreated control) 100% =total destruction Example F2 Pre-Emergence Test Solvent: 5 parts by weight of acetone Emulsifier: 1 part by weight of alkylaryl polyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amount of solvent, the stated amount of emulsifier is added and the concentrate is diluted with water to the desired concentration.\",\n \"Seeds of the test plants are sown in normal soil.\",\n \"After about 24 hours, the soil is sprayed with the preparation of active compound such that the particular amounts of active compound desired are applied per unit area.\",\n \"The concentration of the spray liquor is chosen such that the particular amounts of active compound desired are applied in 1000 l of water/ha.\",\n \"After three weeks, the degree of damage to the plants is rated in % damage in comparison to the development of the untreated control.\",\n \"The figures denote: 0% =no effect (like untreated control) 100% =total destruction Post-emergence/ g.a.i./ Avena Greenhouse ha Alopecurus fatua Setaria Sinapis I-1-a-2 250 90 100 100 80 I-1-a-3 250 90 100 100 80 I-1-c-3 250 100 100 100 80 I-1-a-1 250 100 100 100 70 Post-emergence/ g.a.i/ Sugar- Alope- Avena Echino- Greenhouse ha beet curus fatua chloa Setaria I-2-a-1 250 0 90 95 100 100 Post-emergence/ g.a.i/ Sugar- Avena Echino- Greenhouse ha beet fatua chloa Setaria Sinapis I-1-c-1 250 0 90 100 100 70 Post-emergence/ g.a.i/ Alope- Avena Greenhouse ha curus fatua Setaria Galium I-2-c-1 250 90 100 100 80 pre-emergence/ Soya greenhouse g a.i./ha Wheat bean Digitaria Lolium Setaria Matricana I-1-a-2 60 0 0 100 100 100 95 pre-emergence/ Sugar Avena greenhouse g a.i./ha beet Alopercurus fatua Digitaria Polygonume I-1-a-3 125 0 95 100 100 100 pre-emergence/ g.a.i./ Oilseed Alope- Digi- greenhouse ha rape curus taria Lolium Stellaria I-1-c-2 125 0 95 100 100 100 pre-emergence/ g.a.i./ Soya- Alope- Seta- Stel-a greenhouse ha Wheat bean curus Lolium ria lari I-1-c-3 125 10 0 95 100 100 70 pre-emergence/ g.a.i./ Soya- greenhouse ha bean Lolium Setaria Matricaria Stellaria I-1-a-1 125 0 100 100 100 100 pre-emergence/ g.a.i./ Soya- greenhouse ha bean Bromus Lolium Setaria Abutilon I-2-a-1 125 0 95 100 100 70 Example G In vitro test for the ED50 determination in microorganisms A methanolic solution of the active compound to be tested, admixed with the emulsifier PS16, is pipetted into the wells of microtitre plates.\",\n \"After the solvent has evaporated, 200 μl of potato/dextrose medium are added to each well.\",\n \"Beforehand, a suitable concentration of spores or mycelium of the fungus to be tested was added to the medium.\",\n \"The resulting concentrations of the active compound are 0.1, 1, 10 and 100 ppm.\",\n \"The resulting concentration of the emulsifier is 300 ppm.\",\n \"The plates are then incubated on a shaker at a temperature of 22° C. for 3-5 days, until sufficient growth can be observed in the untreated control.\",\n \"Evaluation is carried out photometrically at a wavelength of 620 nm.\",\n \"The dose of active compound which causes a 50% inhibition of fungal growth compared to the untreated control (ED50) is calculated from the data measured at different concentrations.\",\n \"Active compound Microorganism ED50 value Ex.\",\n \"I-1-a-2 Pyricularia oryzae <0.1 Septoria tritici <0.1 Ustilago avenae <0.1 Ex.\",\n \"I-2-a-2 Pyricularia oryzae 1.96 Septoria tritici <0.1 Ustilago avenae <0.1 Ex.\",\n \"I-2-a-3 Pyricularia oryzae <0.1 Septoria tritici <0.1 Ustilago avenae 0.1 Example H Sphaerotheca Test (Cucumber)/Protective Solvent: 24.5 parts by weight of acetone 24.5 parts by weight of dimethylacetamide Emulsifier: 1 part by weight of alkyl-aryl polyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amounts of solvent and emulsifier, and the concentrate is diluted with water to the desired concentration.\",\n \"To test for protective activity, young plants are sprayed with the preparation of active compound at the stated application rate.\",\n \"After the spray coating has dried on, the plants are inoculated with an aqueous spore suspension of Sphaerotheca fuliginea.\",\n \"The plants are then placed in a greenhouse at about 23° C. and a relative atmospheric humidity of about 70%.\",\n \"Evaluation is carried out 7 days after the inoculation.\",\n \"0% means an efficacy which corresponds to that of the control, whereas an efficacy of 100% means that no infection is observed.\",\n \"Application rate of active Active compound compound in g/ha % efficacy Ex.\",\n \"I-1-a-2 100 100 Example I Venturia Test (Apple)/Protective Solvent: 24.5 parts by weight of acetone 24.5 parts by weight of dimethylacetamide Emulsifier: 1 part by weight of alkyl-aryl polyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amounts of solvent and emulsifier, and the concentrate is diluted with water to the desired concentration.\",\n \"To test for protective activity, young plants are sprayed with the preparation of active compound at the stated application rate.\",\n \"After the spray coating has dried on, the plants are inoculated with an aqueous conidia suspension of the apple scab pathogen Venturia inaequalis and then remain in an incubation cabin at about 20° C. and 100% relative atmospheric humidity for 1 day.\",\n \"The plants are then placed in a greenhouse at about 21° C. and a relative atmospheric humidity of about 90%.\",\n \"Evaluation is carried out 10 days after the inoculation.\",\n \"0% means an efficacy which corresponds to that of the control, whereas an efficacy of 100% means that no infection is observed.\",\n \"Application rate of active Active compound compound in g/ha % efficacy Ex.\",\n \"I-1-a-2 100 91 Example J Alternaria Test (Tomato)/Protective Solvent: 24.5 parts by weight of acetone 24.5 parts by weight of dimethylacetamide Emulsifier: 1 part by weight of alkyl-arylpolyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amounts of solvent and emulsifier, and the concentrate is diluted with water to the desired concentration.\",\n \"To test for protective activity, young plants are sprayed with the preparation of active compound at the stated application rate.\",\n \"After the spray coating has dried on, the plants are inoculated with an aqueous spore suspension of Alternaria solani.\",\n \"The plants are then placed in an incubation cabin at about 200° C. and 100% relative atmospheric humidity.\",\n \"Evaluation is carried out 3 days after the inoculation.\",\n \"0% means an efficacy which corresponds to that of the control, whereas an efficacy of 100% means that no infection is observed.\",\n \"Application rate of active Active compound compound in g/ha % efficacy Ex.\",\n \"I-1-a-2 100 93 Example K Botrytis Test (Bean)/Protective Solvent: 24.5 parts by weight of acetone 24.5 parts by weight of dimethylacetamide Emulsifier: 1 part by weight of alkyl-arylpolyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amounts of solvent and emulsifier, and the concentrate is diluted with water to the desired concentration.\",\n \"To test for protective activity, young plants are sprayed with the preparation of active compound at the stated application rate.\",\n \"After the spray coating has dried on, two small pieces of agar colonized by Botrytis cinerea are placed onto each leaf.\",\n \"The inoculated plants are placed in a dark chamber at about 20° C. and 100% relative atmospheric humidity.\",\n \"2 days after the inoculation, the size of the infested areas on the leaves is evaluated.\",\n \"0% means an efficacy which corresponds to that of the control, whereas an efficacy of 100% means that no infection is observed.\",\n \"Application rate of active Active compound compound in g/ha % efficacy Ex.\",\n \"I-1-a-2 500 92 Example L Critical Concentration Test/Soil Insects-Treatment of Transgenic Plants Test insect: Diabrotica balteata—larvae in soil Solvent: 7 parts by weight of acetone Emulsifier: 1 part by weight of alkylaryl polyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amount of solvent, the stated amount of emulsifier is added and the concentrate is diluted with water to the desired concentration.\",\n \"The preparation of active compound is poured onto the soil.\",\n \"The concentration of the active compound in the preparation is virtually immaterial, only the amount by weight of active compound per volume unit of soil, which is stated in ppm (mg/l) matters.\",\n \"The soil is filled into 0.25 l pots, and these are allowed to stand at 20° C. Immediately after the preparation, 5 pregerminated maize corns of the cultivar YIELD GUARD (trademark of Monsanto Comp., USA) are placed into each pot.\",\n \"After 2 days, the corresponding test insects are placed into the treated soil.\",\n \"After a further 7 days, the efficacy of the active compound is determined by counting the number of maize plants that emerged (1 plant=20% activity).\",\n \"Example M Heliothis Virescens Test—Treatment of Transgenic Plants Solvent: 7 parts by weight of acetone Emulsifier: 1 part by weight of alkylaryl polyglycol ether To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amount of solvent and the stated amount of emulsifier, and the concentrate is diluted with water to the desired concentration.\",\n \"Soya bean shoots (Glycine max) of the cultivar Roundup Ready (trademark of Monsanto Comp., USA) are treated by being dipped into the preparation of active compound of the desired concentration and are populated with the tobacco budworm caterpillar Heliothis virescens while the leaves are still moist.\",\n \"After the desired period of time, the kill of the insects is determined.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10474308"}}},{"rowIdx":798,"cells":{"text":{"kind":"list like","value":[["Reversible life raft comprising removable shelters","Rescue and safety device, permitting the reversibility of a rescue raft, shelter (s)-tent (s), consists of an inflated grille provided with unlockable, spreadable and lockable openings, handles and safety belts.","On this grille are fixed bags (11) containing membranes that expand into the shelter(s) (8) and the floor(s) (7).","The whole is made to be removable, including the weight element, by a fastener system that is laid on two sides of a boat.","The male fasteners are fixed to handles (4) and bags (11), the female fasteners are fixed on the rods and allow, if the raft expands inside out, it to be remounted in place."],["1.Rescue and safety device, permitting the reversibility of a survival raft, shelter (s)-tent (s), consists of an inflated grille, provided with handle and safety belts, whose meshes filled with netting, are provided with openings which can be unlocked, spreaded and locked, said device characterized in that the device is made to be totally reversible by fastening means distributed on two sides of the raft, in that it is composed of means allowing the weight element to fix, stabilize and to make it removable, in that it is composed of at least a bag (11) fixed removably to the grille (1), this bag comprises two membranes, said membranes expand into shelter (8), floor (7) extending on at least a mesh (6) of the grille (1), these membranes-shelters (8) form, by mean of a structure (12), (19), (37), a space which is reinforced by some means and whose sides (13), (18) are also created which have the possibility to expand and fix them to adjacent spaces, contained previously in other bags (11), and to fix also to the bottom of the grille (1), on rods, in order to establish common multiple cell spaces, in that said openings (3) in the netting (2) are openable from over side as well as from underside of the device, by some means, in that said bags (11), handles (4) are provided with fasteners being fixed with fasteners distributed on two sides of the raft, fasteners receiving the periphery of floors and shelters being also distributed on two sides of the raft, in that the bag (s) (11) as well as bases of shelters (8) and floors (7) are composed of ground engaging fasteners (14) with a peg pouch included in said bag, making the shelter (s) (8) with floor (7) independent.","2.Device according to claim 1, characterized in that said membrane forming shelter (8) expands into two parallelepipeds interconnected by a structure in the form of an H (12) with four panels (13) on its sides, said structure is folded following the movement of the center bar of the H which is rotated over half a turn on itself causing the whole of stops/upright posts to be in the form of a bundle with the membrane-shelter and the membrane-floor in front of the latter, being maintained inside a bag, said structure is reinforced when it is filled with water, gas/air or rods, pipes, on the sides of the H, panels of membranes in form of equilateral triangles (13) allow, when the structure is raised, to close sides of a refuge by fixing two said triangles one on the other, of each side, forming also an individual shelter and by fixing the periphery of the base of the shelter which is connected to the grille or to the ground.","3.Device according to claim 1, characterized in that said membranes from at least two bags (11), expand into at least two structures in the form of an H (12) interconnected each time by a center bar of the H which forms a roof, said structures are reinforced when they are filled with water, gas/air or rods, pipes, on the sides of each H, the panels of the membrane in the form of equilateral triangles (13) forming sides allow, when the structure is raised, to close said sides of the refuge, those sides are fixed along one after the other triangle (13) and connecting two triangles (13) of the structure at its end one above the other in order to form a multiple shelter in the form of a tunnel, said parallelepipeds being laid inside form inner raised walls, so that it can be fixed to disengage the space, the periphery of the shelter is fixed to the grille or to the ground.","4.Device according to claim 1, characterized in that a shelter (8) which consists of six structures in the form of an H (12), which expand each time by a center bar of the H forming a roof, said structures are reinforced when they are filled with water, gas/air or rods, pipes, the panels of the membrane in the form of equilateral triangles (13) allow, when the structure is raised, to close sides of the refuge, these sides panels are fixed on the lateral panels along one after the other triangle (13), roofs of the refuge are fixed once over side, once under side of the triangles of square or adjacent rectangular elements in order to establish the whole of several shelters, the inner walls also create of means allowing them to be disengaged, raised, it is the same for the ends used for inlet/outlet openings (38), the base of the shelter is fixed to the grille (1) or to the ground.","5.Device according to claim 1, characterized in that these stops/upright posts (22) consist of folds, which can receive a fluid, rods, pipes.","6.Device according to claim 1, characterized in that said structures in the form of an H (12), form spiders as a pyramid (19), the structure with three sides (37) as well as these membranes-shelters (8), floors (7) are doubled, which are inflated by an automatic air/gas generator or by an air/gas muscle-powered pump (20).","7.Device according to claim 1, characterized in that the surfaces of floors (7) are provided with button holes, slits (25), allowing the passage of handles (4).","8.Device according to claim 1, characterized in that said structures in the form of spiders (19) and said structures consisting of three sides (37) are provided with panels with accordions (17) for its extension to the adjacent space.","9.Device according to claim 1, characterized in that, a bag contains a telescopic mast with a sail and oars."],["<SOH> BACKGROUND ART <EOH>According to the technical background, the object of the document PCT/CH99/00619 is to provide a wider impact surface on the sea wherein an ordinary boat operates and permits someone stuck beneath the device, when it expands, to move onto the device and to stay there by means of unlockable, spreadable and lockable openings permitting shipwrecked persons to grasp the netting in order to rejoin one of the openings.","It also allows, if required, the device to be caught by hand and feet through handles and to use safety belts.","However, survivors are neither protected from the weather influences in cold sea, waves that can pass through the netting, nor the sunlight, and if the raft is opened inside out, shipwrecked persons do not have handles and safety belts.","Also according to the technical background, the document DE 3210 590 A1, 23.03.82 discloses a device for use in open water, which is a multiple place buoy without netting, the tarpaulins of which cover a certain number of openings, and do not form a floor on all the surface.","Shelters are provided, but they are located on certain units, they are fixed and they can't communicate to each other and they form multiple spaces.","Children can slide in the openings, survivors can meet splashes, water can stagnate in the tarpaulins and form pockets there, and when survivors are below the shelter, it is going to create an unbalance and the multiple buoy will be returned, this situation can get worse at the, time of coupling units in a rough sea, because nothing guarantees that shelters are arranged head-to-tail in such way to balance loads.","On the contrary, in the panic condition, unfortunately, it is impossible to perform such operation, many persons must be also exposed to weather influences.","Rescue boats, which are available now in the market, are used only on a small surface, with a very limited number of persons on boards, aircraft glide paths, toboggans and require a skillful folding with a link system in the form of any press-studs so that they are opened in the common direction."],["<SOH> SUMMARY OF THE INVENTION <EOH>Therefore, the object of the invention is to provide a rescue and safety device that is able to overcome these drawbacks.","The advantages of the invention reside in the fact that the rescue raft is made totally reversible by the use of male receiving fasteners distributed on both sides of the raft for forming removable floors, shelters which can be engaged by interconnecting several spaces or a single space at the selected location on the device without the requirement of the proper opening of the raft.","This flexibility is realized by the following features: fasteners distributed on both sides of the boat, removable bags containing shelter and floor, removable handles and safety belts, removable weight element.","By the selection of structures in the form of an H, with triangles, structures with three sides, those sides are provided with panels in the form of bellows, structures in the form of a spider provided with panels in the form of bellows.","The invention will be better understood by reading the description of the embodiments given as non-restrictive examples with regards to the underlying figures."],["TECHNICAL FIELD OF THE INVENTION Rescue and safety device permitting the reversibility of a survival raft, shelter(s)-tent(s), which is to improve survival conditions on a raft, to improve the rescue and safety quality and to create shelters-tents fixed to the raft as well as on shore.","BACKGROUND ART According to the technical background, the object of the document PCT/CH99/00619 is to provide a wider impact surface on the sea wherein an ordinary boat operates and permits someone stuck beneath the device, when it expands, to move onto the device and to stay there by means of unlockable, spreadable and lockable openings permitting shipwrecked persons to grasp the netting in order to rejoin one of the openings.","It also allows, if required, the device to be caught by hand and feet through handles and to use safety belts.","However, survivors are neither protected from the weather influences in cold sea, waves that can pass through the netting, nor the sunlight, and if the raft is opened inside out, shipwrecked persons do not have handles and safety belts.","Also according to the technical background, the document DE 3210 590 A1, 23.03.82 discloses a device for use in open water, which is a multiple place buoy without netting, the tarpaulins of which cover a certain number of openings, and do not form a floor on all the surface.","Shelters are provided, but they are located on certain units, they are fixed and they can't communicate to each other and they form multiple spaces.","Children can slide in the openings, survivors can meet splashes, water can stagnate in the tarpaulins and form pockets there, and when survivors are below the shelter, it is going to create an unbalance and the multiple buoy will be returned, this situation can get worse at the, time of coupling units in a rough sea, because nothing guarantees that shelters are arranged head-to-tail in such way to balance loads.","On the contrary, in the panic condition, unfortunately, it is impossible to perform such operation, many persons must be also exposed to weather influences.","Rescue boats, which are available now in the market, are used only on a small surface, with a very limited number of persons on boards, aircraft glide paths, toboggans and require a skillful folding with a link system in the form of any press-studs so that they are opened in the common direction.","SUMMARY OF THE INVENTION Therefore, the object of the invention is to provide a rescue and safety device that is able to overcome these drawbacks.","The advantages of the invention reside in the fact that the rescue raft is made totally reversible by the use of male receiving fasteners distributed on both sides of the raft for forming removable floors, shelters which can be engaged by interconnecting several spaces or a single space at the selected location on the device without the requirement of the proper opening of the raft.","This flexibility is realized by the following features: fasteners distributed on both sides of the boat, removable bags containing shelter and floor, removable handles and safety belts, removable weight element.","By the selection of structures in the form of an H, with triangles, structures with three sides, those sides are provided with panels in the form of bellows, structures in the form of a spider provided with panels in the form of bellows.","The invention will be better understood by reading the description of the embodiments given as non-restrictive examples with regards to the underlying figures.","BRIEF DESCRIPTION OF FIGURES FIG.","1 shows the grille 1, filled with a netting 2, unlockable, spreadable, and lockable openings 3, handles 4, and safety belts 5.On the side of a mesh 6, it is possible to distinguish a some what elongate open bag 11, which is a membrane folded on itself which forms a raft 8 and an unfolded membrane used as a floor 7 covering four meshes in which there are slits/button holes 25 defined to expose handles 4, the floor membrane fastened to the grille by means of press-studs 9 indicated by points in the drawing.","It is also possible to observe that the raft 8 with inlet and outlet opening 38 is mounted and fastened to the grille by press-studs 9 and a section of floor 7, fixed by press-studs 9, extending on the mesh.","On parts of floors 7, safety belts 5 are undone, they are detached so that they can return onto floors 7 and they can be used.","FIG.","2a shows an apertured tissue strip 27 provided with an accordion 41 in the form of a bellow with a lead 30 in the center of the strip 27, a snap hook 39 and rings 40 at its ends which attach the strip 27 to the rod 52.FIG.","2b shows the weight element 53, the apertured tissue strip 27 with lead 30 in its center, the strip is attached, by means of the snap hook 39, to rings 40 also forming the weigh element 53.It is possible to observe two series of rings distributed in two lines on the rod 52 permitting the placement of the weight element below the raft every time.","FIG.","3a shows a structure 12 in the form of an H, unfolded with its four triangular panels 13.FIG.","3b shows the structure 12 in the form of an H, in haft folded position.","FIG.","4 shows shelters 8, whose two structures 12 in the form of an H are in place, the second structure and two triangular panels 13, one of each side is folded back towards the left, the second parallelepiped of the second structure forming an inner wall, which can not be viewed here and the base of the shelter 8 being fixed to the grille 1 by means of press-studs indicated here by points 9.It is possible to observe here handles 4 and a bag 11 and an inlet/outlet opening 38 provided with zippers indicated by the dash lines 16 on the drawing, as it occurs for the inner wall, with a fastener 31 allowing for disengaging two panels from the wall on sides, the base of the shelter is joined, in this example, to the grille by means of press-studs 9, indicated by points.","FIG.","5 shows a shelter 8 comprising four structures 12 in the form of an H, the fourth structure and two triangles 13, one of each side, are folded back towards the left, forming three inner walls that can be opened and disengaged by zippers 16, the walls cannot be viewed here.","Those structures are joined together to form a tunnel and the base on its periphery is fixed, in this example, to the grille by closing means such as Velcro 10, the location of which is indicated by small points 10.At the end of the tunnel, an inlet/outlet opening 38 is distinguished, which is provided with zippers 16 indicated by dash lines, which permit opening inner walls of the shelter or disengaging the entry upward by the means of an adjustable fastener 42, four structures that occupy four meshes, are provided with safety belts 5.FIG.","6 shows a shelter 8 comprising six structures 12 in the form of an H, two structures at the end of the shelter as well as its two triangles on the sides 13 are folded back from inside towards the left forming three inner walls that can be opened and disengaged by zippers, those walls can not be viewed here, structures are positioned in a rectangular orientation, roofs are joined by fixing triangles one under the other at roofs level, the base of the shelters is fixed by means of zippers 16.Handles 4 and safety belts 5 are provided on the device, it is possible to view the inlet/outlet opening 38 provided with zippers 16 for opening inner walls or disengaging the entry, windows 56, six structures occupying six meshes.","FIG.","7 shows a structure with three sides 37, the fourth is removed from the structure at its top to create an opening with a zipper 16.On the two lateral sides 18, it is possible to distinguish two bellows 17 provided with fasteners 31 permitting the structure to expand in order to connect to the adjacent structure and form a shelter chain, the two vertical lateral panels 18 provided with zippers 16 permitting, when they are opened, panels 18 to be lifted onto these sides and fixed on the shelter from all sides for forming also shelters connected to each other and extending on the surface desired.","It is possible to view handles 4 and unlockable, spreadable and lockable openings 3, it can be observed that safety belts 5 inside the shelter are opened, they were put again on the floor 7 which was stretched and fixed by means of zippers 16, while the other safety belts 5 on the grille are locked, the base of the shelter is fixed to the grille by cords 14 and studs 15 and to upright posts 22 by the Velcro 10.By the transparency it is possible to view the opened bag 11 and the other bag is provided as an example on the device.","FIG.","8 shows a structure in the form of a spider 19 inflated here by means of a muscle-powered pump 20, the pipe 21 of which is extended into the water, and fixed on an upright post by nosepiece 24, the two upright posts 22 are fixed to the grille by the Velcro 10 indicated by small points in the drawing.","On lateral panels 18 there are bellows, in the form of an accordion 17 provided with fasteners 31 to expand the shelter in order to fix two upright posts 22 to the next shelter, one of the sides includes an inlet/outlet opening 38 with a zipper 16, the lateral panels 18 also provided with a zipper 16 as well as the base periphery of the shelter, which is joined to the grille by the same means, these zippers are indicated by dotted lines.","Safety belts 5 are opened within the shelters, ready to be used and they are locked, on the rest of the grille 1, an open bag 11 is provided within the shelter, the other bag on the edge of a mesh and the floor 7 stretched and fixed by a zipper 16 to the grille, cords 14 are provided on rear sides of bags, on the edge of upright posts, shelters, floors (floors are not viewed here).","FIG.","9a shows a sewn tissue strip 23 extended by a seal bead 26.FIG.","9b shows a sewn tissue strip 23 with a fixed seal bead 26 in its surroundings.","FIG.","9c shows the bag 11 with a tube having a slit 32 and a seal bead 26, threaded to the slit 32 of the tube.","FIG.","10a shows a bag 11, on the edge of which fixed sections of the tube 28 are provided, and at the end of which two fasteners 31 are provided.","FIG.","10b shows a rod near the bag 11, at one end of which a handle is provided and at the other end an eye 29 is provided.","FIG.","10c shows a mesh 6 with the netting 2, an unlockable, spreadable and lockable opening 3, handles 4, tube sections 28, which can be flexible or rigid, fixed on the grille for receiving tube sections 28 fixed to the bag 11, see FIG.","10a, adapted to positively engage those fixed to the grille and they will be fixed to each other by the rod 29 in FIG.","10b and the whole will be attached by two fasteners 31 fixed to an end of the bag in FIG.","10a.","FIG.","11a shows a male part fixing mode for bags, handles, a tab 33 provided with a push-button 34.FIG.","11b shows a female receiving piece in the form of semicircle fixed on a support 35 and provided with an orifice 36.FIG.","11c shows the tab 33 engaged in the receiving orifice 35, the push-button 34 being withdrawn from the orifice 36.FIG.","12a shows another male fixing mode for bags, handles, a tab 43 folded on itself with a section to be pressed from above, this tab is provided with two lugs 44.FIG.","12b shows the female half-cone receiving part 45 provided with two orifices 36 into which lugs 44 are engaged for fixing.","FIG.","13 shows a fast locking/opening example with a male part extended by two flexible levers 46, on an axis 57 that can be grasped, with a protruding part 51 adapted to positively engage in orifices, indicated here by hatches 47 of the female part 48, when the pressure is released.","The male part 54 is joined to the female part by a link 55, which prevents the male part from falling into the water.","The female is joined to the netting by a strap 49.DETAILED DESCRIPTION OF THE INVENTION In a non-restrictive example of an embodiment, the present devices are composed mainly of an inflatable grille 1, FIG.","1, which is expanded by means usually used in the inflatable field, whose meshes 6 are fitted with netting 2 which is provided with unlockable, spreadable and lockable opening 3, handles 4 and safety belts 5, which are distributed on the grille 1.In the first step, during shipwrecking and inundation, persons stuck beneath the device, which is expanded, unlock openings 3 and move onto the device, lock these opening, FIG.","1.In the second step, shipwrecked person(s) stabilizes the raft by adjusting the weight element to the desired position, FIG.","12a and FIG.","12b.","If the raft is turned around, safety belts 5 that were attached when the raft is expanded, can be viewed through the netting.","Shipwrecked persons disengage handles 4 and safety belts 5 simultaneously and move firstly onto the device, by meshes of netting 2 if they are wide enough, or by openings 3 which will be opened, or along the fastening of the netting.","Shipwrecked persons fix handles 4 at the new location on the device, disengage an end of the bag, then the other, to pivot it about itself to expose it on the same path, fix it at a location on the device by means of the fastening receiving part 35 or 45, FIG.","11b, 12b, expand the membranes adapted to form a floor 7, FIG.","1, and fix those membranes by means provided for this purpose, such as a Velcro 10, in order to put themselves on a much or less dry place.","In the third step, shipwrecked person(s) organizes the mounting of a shelter 8 or individual or multiple shelters.","A shipwrecked person folds the membrane which is used for the floor 7, captures the structure in the form of spider 19 in this example, FIG.","8, connects muscle pump 20 at it nose piece 24 in the structure, dives the pipe 21 in the water, he can slide it between the floor and the side of the mesh of the grille, he pumps .",".",".","until the structures become rigid, he fixes these two upright posts 22 on the edge of the mesh by the Velcro 10 fixed on the grille, then he climbs, expands the next structure, then disengages bellows 17 held by fasteners 31, releases these two removable upright posts 22 of the first structure to fix them with the Velcro 10 provided on the top part of the second structure, bellows 17 are opened to create the necessary length.","Another shipwrecked person mounted the next shelter, they decide to have more space, lateral panels 18 are raised and fixed one above the other under the roof of the adjacent shelter by the Velcro 10, they fix the base of the shelter with zippers 16 provided on the device.","They are in weather proof condition and can hold handles 4 and reach to safety belts 5.If water enters, they can empty by lifting up a corner of the floor 7.In the hypothesis that they must make landfall, and come to camp on the land, they take away one or several bags 11 and they could establish their camp inside the land territory by means of fasteners, cords 14 distributed on a side of the bag and on the periphery of shelter 8 and the floor 7, FIG.","8.This type of tent which puts in an extended bag is suitable for use with or without rescue boat, in this case, it is not useful to provide ends of the bag with male fasteners, but with cords for fixing posts there, the base of floors, shelters can be fixed sealingly together.","With regard to fastening of panels and floors to rods, fasteners, cords or button holes, pressure straps, straps with knobs, Velcro, snap hooks, sealed zippers, eyes with fasteners can be used, but those are not limited.","Membranes, floor and shelter are fixed to bag in any possible way, they are bonded, heat-sealed, sewn, then rolled, folded on themselves, held by pressure straps, straps with fasteners, Velcro, snap hooks, zippers, cords and knobs, but those are not limited.","The envelope of bags is provided with handle(s), strap(s), Velcro permitting to grasp them better.","Two membranes are interconnected in a bag, they are fixed one after the other, the membrane of the floor is the front and the membrane of the shelter is the rear.","Bags are removably fixed on the grille by means of male receiving fasteners 33 and 43 which are distributed on the front side, and also on the back side of the device.","The female fasteners 35 and 45 are fixed on ends of bags and come to fit with male fasteners.","The male fasteners are distributed on the greater part of or the whole grille for putting on the grille a certain number of bags 11 containing two membranes, floor 7, shelter 8, which make the device lighter because there are not bags all over and there is the choice to set up its tent(s) at location(s) where is desired.","It is the same for handles 4, they are made to be removable by the same kind of male/female fasteners.","Those removable fasteners for bags containing membranes of floors and shelters, are female receiving pieces in the form of a half-cone 35 for example, with an orifice 36.They are fixed on rods.","In receiving pieces, a pull tab, spring tab system with a push-button 34 comes to fit into, for example.","This button is engaged into the orifice and fixes the bag to the grille, FIG.","11a, 11b, 11c.","A variant of an embodiment is a tab folded on itself forming spring 43, with a space for locating the bag, this tab is provided with two lugs 44 which come to engage into two orifices 36 provided for this purpose on female receiving piece fixed on the rod, FIG.","12a, 12b.","Male fasteners are mounted to ends of the bag or for example to ends of a flexible plate strip fixed under the bag, in a variant it can be adjustable in length to accommodate a dimension of the bag, this strip will be threaded in a hem with two opened ends, but this is not limited.","The same type of receiving piece is made for removably fastening handles, two receiving fasteners being bonded, heat-sealed on a plate or mould, the plate with fasteners then being bonded, heat-sealed to rod.","By withdrawing the handle and pressing the button 34 or tab 43, the handle 4 is disengaged from one side then from the other.","This type of fasteners can be adapted for fixing stops, upright posts 22 to the grille.","It is also possible to use clips of safety belts, zippers.","These examples are not limited.","All the fasteners of bags, handles, stops, upright posts of shelters, fasteners of floors will be disposed on two sides of the grille.","Variant: bags can be made to be removable by a fastener composed of a tube provided with a slit 32 fixed along the edge of the bag as receiving piece and of a female seal bead 26 fixed on a tissue piece 23 joined to the grille.","The seal bead 26 is slid by means of a slit inside the tube.","The seal bead can be in an extension of the tissue or at right angles, fixed within surroundings of tissue, FIG.","9a, 9b, 9c.","Variant: bags can be made to be removable by a fastener on the bag 11 with flexible, rigid or elastic tube sections 28, FIG.","10a, in series along the mesh, FIG.","10c, sections of bag and those of the mesh being joined by threading a rod provided with an eye 29, FIG.","10b, with a fastener 31 to fix the whole, FIG.","10a.","Safety belts can be made to be removable by snap hooks fixed to each of its edge, this allows them to be placed end-to-end- and thus creates chains, but this is not limited.","To make a raft reversible, the place of bags and handles on the reverse side of the raft must be indicated someway, this is to save the time for finding and disengaging bags.","The weight element is formed by apertured tissue strips 27 forming sections, within surroundings of each strip, the lead 30, for stabilizing the weight element, and at each end of each strip provided with pleated part in the form of an accordion 41 and means such as rings 40 and snap-hooks 39 allowing the bottom of the strip to hang up/remove on the rod for forming the weight element and releasing it by fixing it to rings on rods 52 by means of snap-hooks.","Rings 40 on the rod 52, are distributed on two lines above and under the raft to ensure that the weight element is always located under the raft or at the necessary position, FIG.","2a, 2b.","Membranes, which are doubled as well as structures, can be connected to one-way valves, if necessary, and inflated automatically or by an air/gas generator or manually with the muscle-powered pump, this is to create an isolation.","Tubular structures are semi-flexible and/or joined since they are folded to range, they can be formed by folds of membranes, be included in folds or connected to the structure.","In folds, hems, it is possible to thread pipes, flexible rods, which can be telescopic, articulatable, bendable, provided with pivot devices to cause them to pivot or interlock, but this is not limited.","Pipes and rods being flexible can be mounted directly just in their manufacture.","If structures are inflated by water, an anti-freeze, which is to be injected, need to be provided as a emergency material.","According to the choice of the user, some rafts are provided with a bag containing a telescopic bar mounted on pivot device, with a sail and oar members, oar member supports can be provided on rods, it is also possible to provide a location for fixing a motor.","The maximal loading and the number of persons will be indicated clearly on the grille.","The length of membranes, floors and/or shelters can be provided to cover one or several meshes, see all the surface of the device or a raft can be made only of two meshes, with a shelter covering a mesh and the floor covering two meshes, but this is not limited, the assembly technique allows the multiplication of spaces by coupling with adjacent spaces, or if preferred, shelters can be formed individually, they are distributed on the device on our desire.","When large meshes, supporting a certain loading, are created, several persons can hold side by side on two rows face to face in the same mesh, the two last persons at each end grasp handles.","Persons who are face to face, with safety belts in addition, are supported feet against feet, when the sea is rough.","The structure in the form of an H applies to parallelepipeds.","The structure on three sides 37, in the form of a square (or rectangular) shown in FIG.","7 also rests on an parallelepiped, membranes of its sides 18 are raised and composed of bellows, in addition, membranes in the form of an accordion 17 which allow panels 18 to withdraw onwards in order to have enough length to be fixed on top of the next shelter.","The structure in the form of a spider in FIG.","8 also is composed of panels 18, whose membrane is raised and a bellow accordion 17 allowing to hang up its upright posts 22 to the next space.","It is not obligatory to fix handles on the side on which bags are fixed.","Between two shelters in the case of a space, clearances, the sealing, will be ensured by fixing strips between upright posts and roofs, for example by Velcro.","On some panels or roofs, transparent membranes forming windows 56 are provided, see FIG.","6.According to the diameter of rods, floors will reach to flush with handles or to rearward to handles, in this case, it must provide also buttonholes/slits 25 to pass through handles 4, FIG.","1.According to the use, rods will be protected from friction by flexible reinforcing strips, used usually in the inflated field.","Unlockable, spreadable and lockable openings 3 are made to be unlockable and lockable from over side as well as from under side, for example by strips of Velcro with a handle on each strip.","Nippers of type pincer, with a spring, long arms, wide and ergonomic that can be grasped, one of the arms of pincer being provided with a link connecting with the netting to avoid the pincer from falling into water, jaws of the pincer surrounding two edges of openings.","By means of a pincer, it is possible to be grasped, of which arms, flexible levers 46 are mounted in one block to the axis 57, each provided with a protruding part 51, these protruding parts 51 of levers come to engage in a female receiving piece 48 which is provided with two orifices 47 to meet two protruding parts 51, this type of pincer can be laid flat, FIG.","13.According to standards and the effective technique for rescue boats, the grille will be constructed by compartments and/or with an inner wall forming an air chamber, these compartments are composed of one or several one-way valves as well as over-pressure valves.","Inflated systems are as usual.","In the case that the builder selects a system with straps for drawing bottles and with several bottles.","Straps are guided by wheels, pulleys, conduits or all other means allowing straps, small cords to slide.","These elements are fixed to rods, straps, small cords are connected to a single handle, loop being viewed from outside of the container.","Bottles are each slid in a protective pouch or fixed by straps or by any usual means.","The grille extends into a container, or into a bag which is opened, for example by yielding the inflation pressure.","According to necessary standards for being certified in countries, the device must be equipped with all means which can ensure the inflation security of the raft and all emergency material usually used to the rescue, cords on rods, ladders, anchor, complete outfitting emergency and rescue kit, accessories, reflecting bands, oar member supports, etc.","A bag, fixed to the grille, with a membrane forming a floor, with or without shelter, is usable to recover a swimming pool and to ensure a sunlight protection."]],"string":"[\n [\n \"Reversible life raft comprising removable shelters\",\n \"Rescue and safety device, permitting the reversibility of a rescue raft, shelter (s)-tent (s), consists of an inflated grille provided with unlockable, spreadable and lockable openings, handles and safety belts.\",\n \"On this grille are fixed bags (11) containing membranes that expand into the shelter(s) (8) and the floor(s) (7).\",\n \"The whole is made to be removable, including the weight element, by a fastener system that is laid on two sides of a boat.\",\n \"The male fasteners are fixed to handles (4) and bags (11), the female fasteners are fixed on the rods and allow, if the raft expands inside out, it to be remounted in place.\"\n ],\n [\n \"1.Rescue and safety device, permitting the reversibility of a survival raft, shelter (s)-tent (s), consists of an inflated grille, provided with handle and safety belts, whose meshes filled with netting, are provided with openings which can be unlocked, spreaded and locked, said device characterized in that the device is made to be totally reversible by fastening means distributed on two sides of the raft, in that it is composed of means allowing the weight element to fix, stabilize and to make it removable, in that it is composed of at least a bag (11) fixed removably to the grille (1), this bag comprises two membranes, said membranes expand into shelter (8), floor (7) extending on at least a mesh (6) of the grille (1), these membranes-shelters (8) form, by mean of a structure (12), (19), (37), a space which is reinforced by some means and whose sides (13), (18) are also created which have the possibility to expand and fix them to adjacent spaces, contained previously in other bags (11), and to fix also to the bottom of the grille (1), on rods, in order to establish common multiple cell spaces, in that said openings (3) in the netting (2) are openable from over side as well as from underside of the device, by some means, in that said bags (11), handles (4) are provided with fasteners being fixed with fasteners distributed on two sides of the raft, fasteners receiving the periphery of floors and shelters being also distributed on two sides of the raft, in that the bag (s) (11) as well as bases of shelters (8) and floors (7) are composed of ground engaging fasteners (14) with a peg pouch included in said bag, making the shelter (s) (8) with floor (7) independent.\",\n \"2.Device according to claim 1, characterized in that said membrane forming shelter (8) expands into two parallelepipeds interconnected by a structure in the form of an H (12) with four panels (13) on its sides, said structure is folded following the movement of the center bar of the H which is rotated over half a turn on itself causing the whole of stops/upright posts to be in the form of a bundle with the membrane-shelter and the membrane-floor in front of the latter, being maintained inside a bag, said structure is reinforced when it is filled with water, gas/air or rods, pipes, on the sides of the H, panels of membranes in form of equilateral triangles (13) allow, when the structure is raised, to close sides of a refuge by fixing two said triangles one on the other, of each side, forming also an individual shelter and by fixing the periphery of the base of the shelter which is connected to the grille or to the ground.\",\n \"3.Device according to claim 1, characterized in that said membranes from at least two bags (11), expand into at least two structures in the form of an H (12) interconnected each time by a center bar of the H which forms a roof, said structures are reinforced when they are filled with water, gas/air or rods, pipes, on the sides of each H, the panels of the membrane in the form of equilateral triangles (13) forming sides allow, when the structure is raised, to close said sides of the refuge, those sides are fixed along one after the other triangle (13) and connecting two triangles (13) of the structure at its end one above the other in order to form a multiple shelter in the form of a tunnel, said parallelepipeds being laid inside form inner raised walls, so that it can be fixed to disengage the space, the periphery of the shelter is fixed to the grille or to the ground.\",\n \"4.Device according to claim 1, characterized in that a shelter (8) which consists of six structures in the form of an H (12), which expand each time by a center bar of the H forming a roof, said structures are reinforced when they are filled with water, gas/air or rods, pipes, the panels of the membrane in the form of equilateral triangles (13) allow, when the structure is raised, to close sides of the refuge, these sides panels are fixed on the lateral panels along one after the other triangle (13), roofs of the refuge are fixed once over side, once under side of the triangles of square or adjacent rectangular elements in order to establish the whole of several shelters, the inner walls also create of means allowing them to be disengaged, raised, it is the same for the ends used for inlet/outlet openings (38), the base of the shelter is fixed to the grille (1) or to the ground.\",\n \"5.Device according to claim 1, characterized in that these stops/upright posts (22) consist of folds, which can receive a fluid, rods, pipes.\",\n \"6.Device according to claim 1, characterized in that said structures in the form of an H (12), form spiders as a pyramid (19), the structure with three sides (37) as well as these membranes-shelters (8), floors (7) are doubled, which are inflated by an automatic air/gas generator or by an air/gas muscle-powered pump (20).\",\n \"7.Device according to claim 1, characterized in that the surfaces of floors (7) are provided with button holes, slits (25), allowing the passage of handles (4).\",\n \"8.Device according to claim 1, characterized in that said structures in the form of spiders (19) and said structures consisting of three sides (37) are provided with panels with accordions (17) for its extension to the adjacent space.\",\n \"9.Device according to claim 1, characterized in that, a bag contains a telescopic mast with a sail and oars.\"\n ],\n [\n \"<SOH> BACKGROUND ART <EOH>According to the technical background, the object of the document PCT/CH99/00619 is to provide a wider impact surface on the sea wherein an ordinary boat operates and permits someone stuck beneath the device, when it expands, to move onto the device and to stay there by means of unlockable, spreadable and lockable openings permitting shipwrecked persons to grasp the netting in order to rejoin one of the openings.\",\n \"It also allows, if required, the device to be caught by hand and feet through handles and to use safety belts.\",\n \"However, survivors are neither protected from the weather influences in cold sea, waves that can pass through the netting, nor the sunlight, and if the raft is opened inside out, shipwrecked persons do not have handles and safety belts.\",\n \"Also according to the technical background, the document DE 3210 590 A1, 23.03.82 discloses a device for use in open water, which is a multiple place buoy without netting, the tarpaulins of which cover a certain number of openings, and do not form a floor on all the surface.\",\n \"Shelters are provided, but they are located on certain units, they are fixed and they can't communicate to each other and they form multiple spaces.\",\n \"Children can slide in the openings, survivors can meet splashes, water can stagnate in the tarpaulins and form pockets there, and when survivors are below the shelter, it is going to create an unbalance and the multiple buoy will be returned, this situation can get worse at the, time of coupling units in a rough sea, because nothing guarantees that shelters are arranged head-to-tail in such way to balance loads.\",\n \"On the contrary, in the panic condition, unfortunately, it is impossible to perform such operation, many persons must be also exposed to weather influences.\",\n \"Rescue boats, which are available now in the market, are used only on a small surface, with a very limited number of persons on boards, aircraft glide paths, toboggans and require a skillful folding with a link system in the form of any press-studs so that they are opened in the common direction.\"\n ],\n [\n \"<SOH> SUMMARY OF THE INVENTION <EOH>Therefore, the object of the invention is to provide a rescue and safety device that is able to overcome these drawbacks.\",\n \"The advantages of the invention reside in the fact that the rescue raft is made totally reversible by the use of male receiving fasteners distributed on both sides of the raft for forming removable floors, shelters which can be engaged by interconnecting several spaces or a single space at the selected location on the device without the requirement of the proper opening of the raft.\",\n \"This flexibility is realized by the following features: fasteners distributed on both sides of the boat, removable bags containing shelter and floor, removable handles and safety belts, removable weight element.\",\n \"By the selection of structures in the form of an H, with triangles, structures with three sides, those sides are provided with panels in the form of bellows, structures in the form of a spider provided with panels in the form of bellows.\",\n \"The invention will be better understood by reading the description of the embodiments given as non-restrictive examples with regards to the underlying figures.\"\n ],\n [\n \"TECHNICAL FIELD OF THE INVENTION Rescue and safety device permitting the reversibility of a survival raft, shelter(s)-tent(s), which is to improve survival conditions on a raft, to improve the rescue and safety quality and to create shelters-tents fixed to the raft as well as on shore.\",\n \"BACKGROUND ART According to the technical background, the object of the document PCT/CH99/00619 is to provide a wider impact surface on the sea wherein an ordinary boat operates and permits someone stuck beneath the device, when it expands, to move onto the device and to stay there by means of unlockable, spreadable and lockable openings permitting shipwrecked persons to grasp the netting in order to rejoin one of the openings.\",\n \"It also allows, if required, the device to be caught by hand and feet through handles and to use safety belts.\",\n \"However, survivors are neither protected from the weather influences in cold sea, waves that can pass through the netting, nor the sunlight, and if the raft is opened inside out, shipwrecked persons do not have handles and safety belts.\",\n \"Also according to the technical background, the document DE 3210 590 A1, 23.03.82 discloses a device for use in open water, which is a multiple place buoy without netting, the tarpaulins of which cover a certain number of openings, and do not form a floor on all the surface.\",\n \"Shelters are provided, but they are located on certain units, they are fixed and they can't communicate to each other and they form multiple spaces.\",\n \"Children can slide in the openings, survivors can meet splashes, water can stagnate in the tarpaulins and form pockets there, and when survivors are below the shelter, it is going to create an unbalance and the multiple buoy will be returned, this situation can get worse at the, time of coupling units in a rough sea, because nothing guarantees that shelters are arranged head-to-tail in such way to balance loads.\",\n \"On the contrary, in the panic condition, unfortunately, it is impossible to perform such operation, many persons must be also exposed to weather influences.\",\n \"Rescue boats, which are available now in the market, are used only on a small surface, with a very limited number of persons on boards, aircraft glide paths, toboggans and require a skillful folding with a link system in the form of any press-studs so that they are opened in the common direction.\",\n \"SUMMARY OF THE INVENTION Therefore, the object of the invention is to provide a rescue and safety device that is able to overcome these drawbacks.\",\n \"The advantages of the invention reside in the fact that the rescue raft is made totally reversible by the use of male receiving fasteners distributed on both sides of the raft for forming removable floors, shelters which can be engaged by interconnecting several spaces or a single space at the selected location on the device without the requirement of the proper opening of the raft.\",\n \"This flexibility is realized by the following features: fasteners distributed on both sides of the boat, removable bags containing shelter and floor, removable handles and safety belts, removable weight element.\",\n \"By the selection of structures in the form of an H, with triangles, structures with three sides, those sides are provided with panels in the form of bellows, structures in the form of a spider provided with panels in the form of bellows.\",\n \"The invention will be better understood by reading the description of the embodiments given as non-restrictive examples with regards to the underlying figures.\",\n \"BRIEF DESCRIPTION OF FIGURES FIG.\",\n \"1 shows the grille 1, filled with a netting 2, unlockable, spreadable, and lockable openings 3, handles 4, and safety belts 5.On the side of a mesh 6, it is possible to distinguish a some what elongate open bag 11, which is a membrane folded on itself which forms a raft 8 and an unfolded membrane used as a floor 7 covering four meshes in which there are slits/button holes 25 defined to expose handles 4, the floor membrane fastened to the grille by means of press-studs 9 indicated by points in the drawing.\",\n \"It is also possible to observe that the raft 8 with inlet and outlet opening 38 is mounted and fastened to the grille by press-studs 9 and a section of floor 7, fixed by press-studs 9, extending on the mesh.\",\n \"On parts of floors 7, safety belts 5 are undone, they are detached so that they can return onto floors 7 and they can be used.\",\n \"FIG.\",\n \"2a shows an apertured tissue strip 27 provided with an accordion 41 in the form of a bellow with a lead 30 in the center of the strip 27, a snap hook 39 and rings 40 at its ends which attach the strip 27 to the rod 52.FIG.\",\n \"2b shows the weight element 53, the apertured tissue strip 27 with lead 30 in its center, the strip is attached, by means of the snap hook 39, to rings 40 also forming the weigh element 53.It is possible to observe two series of rings distributed in two lines on the rod 52 permitting the placement of the weight element below the raft every time.\",\n \"FIG.\",\n \"3a shows a structure 12 in the form of an H, unfolded with its four triangular panels 13.FIG.\",\n \"3b shows the structure 12 in the form of an H, in haft folded position.\",\n \"FIG.\",\n \"4 shows shelters 8, whose two structures 12 in the form of an H are in place, the second structure and two triangular panels 13, one of each side is folded back towards the left, the second parallelepiped of the second structure forming an inner wall, which can not be viewed here and the base of the shelter 8 being fixed to the grille 1 by means of press-studs indicated here by points 9.It is possible to observe here handles 4 and a bag 11 and an inlet/outlet opening 38 provided with zippers indicated by the dash lines 16 on the drawing, as it occurs for the inner wall, with a fastener 31 allowing for disengaging two panels from the wall on sides, the base of the shelter is joined, in this example, to the grille by means of press-studs 9, indicated by points.\",\n \"FIG.\",\n \"5 shows a shelter 8 comprising four structures 12 in the form of an H, the fourth structure and two triangles 13, one of each side, are folded back towards the left, forming three inner walls that can be opened and disengaged by zippers 16, the walls cannot be viewed here.\",\n \"Those structures are joined together to form a tunnel and the base on its periphery is fixed, in this example, to the grille by closing means such as Velcro 10, the location of which is indicated by small points 10.At the end of the tunnel, an inlet/outlet opening 38 is distinguished, which is provided with zippers 16 indicated by dash lines, which permit opening inner walls of the shelter or disengaging the entry upward by the means of an adjustable fastener 42, four structures that occupy four meshes, are provided with safety belts 5.FIG.\",\n \"6 shows a shelter 8 comprising six structures 12 in the form of an H, two structures at the end of the shelter as well as its two triangles on the sides 13 are folded back from inside towards the left forming three inner walls that can be opened and disengaged by zippers, those walls can not be viewed here, structures are positioned in a rectangular orientation, roofs are joined by fixing triangles one under the other at roofs level, the base of the shelters is fixed by means of zippers 16.Handles 4 and safety belts 5 are provided on the device, it is possible to view the inlet/outlet opening 38 provided with zippers 16 for opening inner walls or disengaging the entry, windows 56, six structures occupying six meshes.\",\n \"FIG.\",\n \"7 shows a structure with three sides 37, the fourth is removed from the structure at its top to create an opening with a zipper 16.On the two lateral sides 18, it is possible to distinguish two bellows 17 provided with fasteners 31 permitting the structure to expand in order to connect to the adjacent structure and form a shelter chain, the two vertical lateral panels 18 provided with zippers 16 permitting, when they are opened, panels 18 to be lifted onto these sides and fixed on the shelter from all sides for forming also shelters connected to each other and extending on the surface desired.\",\n \"It is possible to view handles 4 and unlockable, spreadable and lockable openings 3, it can be observed that safety belts 5 inside the shelter are opened, they were put again on the floor 7 which was stretched and fixed by means of zippers 16, while the other safety belts 5 on the grille are locked, the base of the shelter is fixed to the grille by cords 14 and studs 15 and to upright posts 22 by the Velcro 10.By the transparency it is possible to view the opened bag 11 and the other bag is provided as an example on the device.\",\n \"FIG.\",\n \"8 shows a structure in the form of a spider 19 inflated here by means of a muscle-powered pump 20, the pipe 21 of which is extended into the water, and fixed on an upright post by nosepiece 24, the two upright posts 22 are fixed to the grille by the Velcro 10 indicated by small points in the drawing.\",\n \"On lateral panels 18 there are bellows, in the form of an accordion 17 provided with fasteners 31 to expand the shelter in order to fix two upright posts 22 to the next shelter, one of the sides includes an inlet/outlet opening 38 with a zipper 16, the lateral panels 18 also provided with a zipper 16 as well as the base periphery of the shelter, which is joined to the grille by the same means, these zippers are indicated by dotted lines.\",\n \"Safety belts 5 are opened within the shelters, ready to be used and they are locked, on the rest of the grille 1, an open bag 11 is provided within the shelter, the other bag on the edge of a mesh and the floor 7 stretched and fixed by a zipper 16 to the grille, cords 14 are provided on rear sides of bags, on the edge of upright posts, shelters, floors (floors are not viewed here).\",\n \"FIG.\",\n \"9a shows a sewn tissue strip 23 extended by a seal bead 26.FIG.\",\n \"9b shows a sewn tissue strip 23 with a fixed seal bead 26 in its surroundings.\",\n \"FIG.\",\n \"9c shows the bag 11 with a tube having a slit 32 and a seal bead 26, threaded to the slit 32 of the tube.\",\n \"FIG.\",\n \"10a shows a bag 11, on the edge of which fixed sections of the tube 28 are provided, and at the end of which two fasteners 31 are provided.\",\n \"FIG.\",\n \"10b shows a rod near the bag 11, at one end of which a handle is provided and at the other end an eye 29 is provided.\",\n \"FIG.\",\n \"10c shows a mesh 6 with the netting 2, an unlockable, spreadable and lockable opening 3, handles 4, tube sections 28, which can be flexible or rigid, fixed on the grille for receiving tube sections 28 fixed to the bag 11, see FIG.\",\n \"10a, adapted to positively engage those fixed to the grille and they will be fixed to each other by the rod 29 in FIG.\",\n \"10b and the whole will be attached by two fasteners 31 fixed to an end of the bag in FIG.\",\n \"10a.\",\n \"FIG.\",\n \"11a shows a male part fixing mode for bags, handles, a tab 33 provided with a push-button 34.FIG.\",\n \"11b shows a female receiving piece in the form of semicircle fixed on a support 35 and provided with an orifice 36.FIG.\",\n \"11c shows the tab 33 engaged in the receiving orifice 35, the push-button 34 being withdrawn from the orifice 36.FIG.\",\n \"12a shows another male fixing mode for bags, handles, a tab 43 folded on itself with a section to be pressed from above, this tab is provided with two lugs 44.FIG.\",\n \"12b shows the female half-cone receiving part 45 provided with two orifices 36 into which lugs 44 are engaged for fixing.\",\n \"FIG.\",\n \"13 shows a fast locking/opening example with a male part extended by two flexible levers 46, on an axis 57 that can be grasped, with a protruding part 51 adapted to positively engage in orifices, indicated here by hatches 47 of the female part 48, when the pressure is released.\",\n \"The male part 54 is joined to the female part by a link 55, which prevents the male part from falling into the water.\",\n \"The female is joined to the netting by a strap 49.DETAILED DESCRIPTION OF THE INVENTION In a non-restrictive example of an embodiment, the present devices are composed mainly of an inflatable grille 1, FIG.\",\n \"1, which is expanded by means usually used in the inflatable field, whose meshes 6 are fitted with netting 2 which is provided with unlockable, spreadable and lockable opening 3, handles 4 and safety belts 5, which are distributed on the grille 1.In the first step, during shipwrecking and inundation, persons stuck beneath the device, which is expanded, unlock openings 3 and move onto the device, lock these opening, FIG.\",\n \"1.In the second step, shipwrecked person(s) stabilizes the raft by adjusting the weight element to the desired position, FIG.\",\n \"12a and FIG.\",\n \"12b.\",\n \"If the raft is turned around, safety belts 5 that were attached when the raft is expanded, can be viewed through the netting.\",\n \"Shipwrecked persons disengage handles 4 and safety belts 5 simultaneously and move firstly onto the device, by meshes of netting 2 if they are wide enough, or by openings 3 which will be opened, or along the fastening of the netting.\",\n \"Shipwrecked persons fix handles 4 at the new location on the device, disengage an end of the bag, then the other, to pivot it about itself to expose it on the same path, fix it at a location on the device by means of the fastening receiving part 35 or 45, FIG.\",\n \"11b, 12b, expand the membranes adapted to form a floor 7, FIG.\",\n \"1, and fix those membranes by means provided for this purpose, such as a Velcro 10, in order to put themselves on a much or less dry place.\",\n \"In the third step, shipwrecked person(s) organizes the mounting of a shelter 8 or individual or multiple shelters.\",\n \"A shipwrecked person folds the membrane which is used for the floor 7, captures the structure in the form of spider 19 in this example, FIG.\",\n \"8, connects muscle pump 20 at it nose piece 24 in the structure, dives the pipe 21 in the water, he can slide it between the floor and the side of the mesh of the grille, he pumps .\",\n \".\",\n \".\",\n \"until the structures become rigid, he fixes these two upright posts 22 on the edge of the mesh by the Velcro 10 fixed on the grille, then he climbs, expands the next structure, then disengages bellows 17 held by fasteners 31, releases these two removable upright posts 22 of the first structure to fix them with the Velcro 10 provided on the top part of the second structure, bellows 17 are opened to create the necessary length.\",\n \"Another shipwrecked person mounted the next shelter, they decide to have more space, lateral panels 18 are raised and fixed one above the other under the roof of the adjacent shelter by the Velcro 10, they fix the base of the shelter with zippers 16 provided on the device.\",\n \"They are in weather proof condition and can hold handles 4 and reach to safety belts 5.If water enters, they can empty by lifting up a corner of the floor 7.In the hypothesis that they must make landfall, and come to camp on the land, they take away one or several bags 11 and they could establish their camp inside the land territory by means of fasteners, cords 14 distributed on a side of the bag and on the periphery of shelter 8 and the floor 7, FIG.\",\n \"8.This type of tent which puts in an extended bag is suitable for use with or without rescue boat, in this case, it is not useful to provide ends of the bag with male fasteners, but with cords for fixing posts there, the base of floors, shelters can be fixed sealingly together.\",\n \"With regard to fastening of panels and floors to rods, fasteners, cords or button holes, pressure straps, straps with knobs, Velcro, snap hooks, sealed zippers, eyes with fasteners can be used, but those are not limited.\",\n \"Membranes, floor and shelter are fixed to bag in any possible way, they are bonded, heat-sealed, sewn, then rolled, folded on themselves, held by pressure straps, straps with fasteners, Velcro, snap hooks, zippers, cords and knobs, but those are not limited.\",\n \"The envelope of bags is provided with handle(s), strap(s), Velcro permitting to grasp them better.\",\n \"Two membranes are interconnected in a bag, they are fixed one after the other, the membrane of the floor is the front and the membrane of the shelter is the rear.\",\n \"Bags are removably fixed on the grille by means of male receiving fasteners 33 and 43 which are distributed on the front side, and also on the back side of the device.\",\n \"The female fasteners 35 and 45 are fixed on ends of bags and come to fit with male fasteners.\",\n \"The male fasteners are distributed on the greater part of or the whole grille for putting on the grille a certain number of bags 11 containing two membranes, floor 7, shelter 8, which make the device lighter because there are not bags all over and there is the choice to set up its tent(s) at location(s) where is desired.\",\n \"It is the same for handles 4, they are made to be removable by the same kind of male/female fasteners.\",\n \"Those removable fasteners for bags containing membranes of floors and shelters, are female receiving pieces in the form of a half-cone 35 for example, with an orifice 36.They are fixed on rods.\",\n \"In receiving pieces, a pull tab, spring tab system with a push-button 34 comes to fit into, for example.\",\n \"This button is engaged into the orifice and fixes the bag to the grille, FIG.\",\n \"11a, 11b, 11c.\",\n \"A variant of an embodiment is a tab folded on itself forming spring 43, with a space for locating the bag, this tab is provided with two lugs 44 which come to engage into two orifices 36 provided for this purpose on female receiving piece fixed on the rod, FIG.\",\n \"12a, 12b.\",\n \"Male fasteners are mounted to ends of the bag or for example to ends of a flexible plate strip fixed under the bag, in a variant it can be adjustable in length to accommodate a dimension of the bag, this strip will be threaded in a hem with two opened ends, but this is not limited.\",\n \"The same type of receiving piece is made for removably fastening handles, two receiving fasteners being bonded, heat-sealed on a plate or mould, the plate with fasteners then being bonded, heat-sealed to rod.\",\n \"By withdrawing the handle and pressing the button 34 or tab 43, the handle 4 is disengaged from one side then from the other.\",\n \"This type of fasteners can be adapted for fixing stops, upright posts 22 to the grille.\",\n \"It is also possible to use clips of safety belts, zippers.\",\n \"These examples are not limited.\",\n \"All the fasteners of bags, handles, stops, upright posts of shelters, fasteners of floors will be disposed on two sides of the grille.\",\n \"Variant: bags can be made to be removable by a fastener composed of a tube provided with a slit 32 fixed along the edge of the bag as receiving piece and of a female seal bead 26 fixed on a tissue piece 23 joined to the grille.\",\n \"The seal bead 26 is slid by means of a slit inside the tube.\",\n \"The seal bead can be in an extension of the tissue or at right angles, fixed within surroundings of tissue, FIG.\",\n \"9a, 9b, 9c.\",\n \"Variant: bags can be made to be removable by a fastener on the bag 11 with flexible, rigid or elastic tube sections 28, FIG.\",\n \"10a, in series along the mesh, FIG.\",\n \"10c, sections of bag and those of the mesh being joined by threading a rod provided with an eye 29, FIG.\",\n \"10b, with a fastener 31 to fix the whole, FIG.\",\n \"10a.\",\n \"Safety belts can be made to be removable by snap hooks fixed to each of its edge, this allows them to be placed end-to-end- and thus creates chains, but this is not limited.\",\n \"To make a raft reversible, the place of bags and handles on the reverse side of the raft must be indicated someway, this is to save the time for finding and disengaging bags.\",\n \"The weight element is formed by apertured tissue strips 27 forming sections, within surroundings of each strip, the lead 30, for stabilizing the weight element, and at each end of each strip provided with pleated part in the form of an accordion 41 and means such as rings 40 and snap-hooks 39 allowing the bottom of the strip to hang up/remove on the rod for forming the weight element and releasing it by fixing it to rings on rods 52 by means of snap-hooks.\",\n \"Rings 40 on the rod 52, are distributed on two lines above and under the raft to ensure that the weight element is always located under the raft or at the necessary position, FIG.\",\n \"2a, 2b.\",\n \"Membranes, which are doubled as well as structures, can be connected to one-way valves, if necessary, and inflated automatically or by an air/gas generator or manually with the muscle-powered pump, this is to create an isolation.\",\n \"Tubular structures are semi-flexible and/or joined since they are folded to range, they can be formed by folds of membranes, be included in folds or connected to the structure.\",\n \"In folds, hems, it is possible to thread pipes, flexible rods, which can be telescopic, articulatable, bendable, provided with pivot devices to cause them to pivot or interlock, but this is not limited.\",\n \"Pipes and rods being flexible can be mounted directly just in their manufacture.\",\n \"If structures are inflated by water, an anti-freeze, which is to be injected, need to be provided as a emergency material.\",\n \"According to the choice of the user, some rafts are provided with a bag containing a telescopic bar mounted on pivot device, with a sail and oar members, oar member supports can be provided on rods, it is also possible to provide a location for fixing a motor.\",\n \"The maximal loading and the number of persons will be indicated clearly on the grille.\",\n \"The length of membranes, floors and/or shelters can be provided to cover one or several meshes, see all the surface of the device or a raft can be made only of two meshes, with a shelter covering a mesh and the floor covering two meshes, but this is not limited, the assembly technique allows the multiplication of spaces by coupling with adjacent spaces, or if preferred, shelters can be formed individually, they are distributed on the device on our desire.\",\n \"When large meshes, supporting a certain loading, are created, several persons can hold side by side on two rows face to face in the same mesh, the two last persons at each end grasp handles.\",\n \"Persons who are face to face, with safety belts in addition, are supported feet against feet, when the sea is rough.\",\n \"The structure in the form of an H applies to parallelepipeds.\",\n \"The structure on three sides 37, in the form of a square (or rectangular) shown in FIG.\",\n \"7 also rests on an parallelepiped, membranes of its sides 18 are raised and composed of bellows, in addition, membranes in the form of an accordion 17 which allow panels 18 to withdraw onwards in order to have enough length to be fixed on top of the next shelter.\",\n \"The structure in the form of a spider in FIG.\",\n \"8 also is composed of panels 18, whose membrane is raised and a bellow accordion 17 allowing to hang up its upright posts 22 to the next space.\",\n \"It is not obligatory to fix handles on the side on which bags are fixed.\",\n \"Between two shelters in the case of a space, clearances, the sealing, will be ensured by fixing strips between upright posts and roofs, for example by Velcro.\",\n \"On some panels or roofs, transparent membranes forming windows 56 are provided, see FIG.\",\n \"6.According to the diameter of rods, floors will reach to flush with handles or to rearward to handles, in this case, it must provide also buttonholes/slits 25 to pass through handles 4, FIG.\",\n \"1.According to the use, rods will be protected from friction by flexible reinforcing strips, used usually in the inflated field.\",\n \"Unlockable, spreadable and lockable openings 3 are made to be unlockable and lockable from over side as well as from under side, for example by strips of Velcro with a handle on each strip.\",\n \"Nippers of type pincer, with a spring, long arms, wide and ergonomic that can be grasped, one of the arms of pincer being provided with a link connecting with the netting to avoid the pincer from falling into water, jaws of the pincer surrounding two edges of openings.\",\n \"By means of a pincer, it is possible to be grasped, of which arms, flexible levers 46 are mounted in one block to the axis 57, each provided with a protruding part 51, these protruding parts 51 of levers come to engage in a female receiving piece 48 which is provided with two orifices 47 to meet two protruding parts 51, this type of pincer can be laid flat, FIG.\",\n \"13.According to standards and the effective technique for rescue boats, the grille will be constructed by compartments and/or with an inner wall forming an air chamber, these compartments are composed of one or several one-way valves as well as over-pressure valves.\",\n \"Inflated systems are as usual.\",\n \"In the case that the builder selects a system with straps for drawing bottles and with several bottles.\",\n \"Straps are guided by wheels, pulleys, conduits or all other means allowing straps, small cords to slide.\",\n \"These elements are fixed to rods, straps, small cords are connected to a single handle, loop being viewed from outside of the container.\",\n \"Bottles are each slid in a protective pouch or fixed by straps or by any usual means.\",\n \"The grille extends into a container, or into a bag which is opened, for example by yielding the inflation pressure.\",\n \"According to necessary standards for being certified in countries, the device must be equipped with all means which can ensure the inflation security of the raft and all emergency material usually used to the rescue, cords on rods, ladders, anchor, complete outfitting emergency and rescue kit, accessories, reflecting bands, oar member supports, etc.\",\n \"A bag, fixed to the grille, with a membrane forming a floor, with or without shelter, is usable to recover a swimming pool and to ensure a sunlight protection.\"\n ]\n]","isConversational":false},"source":{"kind":"string","value":"Patent_10474317"}}},{"rowIdx":799,"cells":{"text":{"kind":"list like","value":[["System for finding vehicles parked inside buildings and roofed surfaces","To find an automobile or another vehicle that it was left parked, in spaces intended for such purpose, by means of widely used current technology, of simple use and without adding accessories to the vehicle.","Several devices, characterized by their modular design, the flat relative cartesian coordinates and computer programs, constitute the system jointly, that orienting and guiding the user to the parking space of his/her vehicle, who does not have to worry about its location.","This invention uses the following constituents: presentation box, programmed microcontroller, audiovisual electronic alerting unit (7), Smartcard connector (5), barcode laser scanner (8), LCD screen (6), power source, Smartcard (2.1), ticket-card with bar code (2.2), coordinates and computer programs.","The “Registering Post” (1.1) defines and registers the localization of the parked vehicle and the “Orienting Post” (1.2) visualizes the information in its LCD screen (6) to orient and guide the user, until finding his/her parked vehicle."],["1.Systems for Finding Vehicles Parked Inside Buildings and Roofed Surfaces, characterized by having got two types of modules (1), that can contain a Smartcard connector, a bar code laser scanner, a retro-illuminated LCD screen, a languages selector, an audiovisual electronic alerting unit, a power source connected to the general net of electric fixtures, a microcontroller integrated in the electronic circuit and a presentation box.","2.The system according to the preceding claim, characterized by having got two types of modules (1): a) the “Registering Post” (1.1) allows to register or store in the data support (2) used by the parking, the localization code of the parked vehicle, being necessary its installation only in parking areas.","b) the “Orienting Post” (1.2) allows to orient and guide the user until finding his/her parked vehicle, based on the lecturing registration of the data support (2), being able to install in accesses, entries, corridors, stairways, parking areas, and so on, where the quantity of these modules will be condicionated by the optimum service.","3.The system according to the preceding claims, characterized because the “Registering Post” (3) has got to use two types of data support (2) in the localization register of the parked vehicle: a) the Smartcard (2.1), that is electronically recording, when it is inserted in the connector (5).","b) ticket-card of bar code (2.2), that must be remove from recipient (3.2) puts for such effect.","c) magnetic band card where the bar code (2.2) is registered, by means of the stamp of automatic ink (simile Trodat), pressing it by hand.","(3.2).","4.The system according to the preceding claims, characterized because the “Orienting Post” (4) has got to recover the localization of the parked vehicle, registered from the data support (2), of two types of lectors: a) Smartcard connector (5) being activated the reading, when the Smartcard (2.1) is inserted.","b) laser scanner (8) being activated the lecturing, when the magnetic band card (2.2) passes by its influence area, or the ticket-card (2.2) where the bar code was registered, according to the used format.","5.The system according to the preceding claims, characterized because the “Post Orienting” (4) has got a retro-illuminated LCD screen (6), with an alphanumeric text to visualize the information that allows to orient and guide the user that requests it, until finding his/her parked vehicle.","6.The system according to the preceding claims, characterized because the two types of modules (1), to identify what type of process is executing by the microcontroller, they have got the audiovisual electronic alerting unit (7), which is compounded by a visual colour sign and quantity of Led (7.1) and an acoustic sign taken place by the loudspeaker (7.2).","7.The system according to the preceding claims, characterized because the two types of modules (1), have got a microcontroller integrated in an electronic circuit, which controls all the processes that are executed in the system, by means of a structured computer program and designed especially to develop the necessary functions and that it is registered in the microcontroller memory, when the system is installed.","8.The system according to the preceding claims, characterized because the computer program that is installed in the microcontroller, it mainly controls the following processes: a) the manage system of flat relative cartesian coordinates.","b) the recording and lecturing in the Smartcard of the localization code of the parked vehicle.","c) the data admission sent by the laser scanner, corresponding to the lecturing of cards with bar code d) the visualization in the LCD screen of the different informations.","e) the control of the audiovisual electronic alerting unit f) reset the system g) the lecturing and memorization of the “Post” coordinates h) the auto-checking of the “Post” correct initiation i) the control of the different devices correct operations of the system j) the control of the language change in which the informations are visualized.","9.The system according to the preceding claims, characterized by using a system of flat relative cartesian coordinates, to define the localization of each parked vehicle, which they are own and unique of each parking space intended for parking inside the same building or space used for parking.","10.The system according to the preceding claims, characterized because in the system of flat relative cartesian coordinates, the difference in the axis “z” indicates the quantity of floors or plants expressed in unities which it must be up or down, to get to the right place, the difference in the axis “x” and in the sense that the LCD screen indicates, it expresses the quantity of meters the parked vehicle is, according to the walking by the street of the “Orienting Post” and the difference in the axis “y” expresses the quantity of meters the parked vehicle is far from the previous street to the right or to the left, as it is the difference in the axis “y” positive or negative.","11.The system according to the preceding claims, characterized because the Smartcard (2.1), used as data support (2), it isn't personalized to a parking in exclusiveness, being able to use the same card, in all the parkings or parking that have installed the system of this invention.","12.The system according to the preceding claims, characterized because the “Orienting Post” possesses a series of buttons that allow to choose the language in which we want to visualize the informations in the LCD screen, pressing once the corresponding to the selected language."],["<SOH> BACKGROUND OF THE INVENTION <EOH>In the current parking, the only existing method to locate a parked vehicle later on, It is the identification of the parking space by means of the letters and digits combination, for example J 320 .","It must be usually completed with the number of the floor or plant and the sector inside the floor or plant, being the localization as follows: floor 3 , sector Cinema, street J, parking space 320 .","The range of colors, is used to delimit the different parking areas.","A form to palliate this problem, is that the user takes note of the code, either memorizing, recording or registering it electronically, or writing it in any element at his/her disposal, what means that he/she will have to transport some extra element in his/her pocket.","In these places any method that is based on the Global Position System (GPS), GLONASS or similar as the next Galilean lacks usefulness, because there are some obstacles (the own building) that interfere in the satellites signs which must be received by the navigator or receiving GPS installed in the vehicle, to define the exact position in coordinates, at the moment he/she leaves it parked.","When the user wants to find his/her parked vehicle, he/she has the same inconvenience with the receiving portable GPS, where previously it was also impossible to register the localization of the parking space in question.","Also this coordinates used to have a short coming of at least 40 meters, unless it arrange differential phase or the DGPS corrections.","In the market there are different optic systems and/or mechanics and/or electronic and/or electromagnetic which solve the problem in question, but their biggest inconvenience reside in the high cost of installation and maintenance, due to its complexity, sophistication and the human personnel who must be devoted to be of that service.","These systems can have interferences of different characteristics which diminish their effectiveness in the supposed solution, and which can transform them in unviable or little practical.","It can consider that some of them attempt against the anonymity and people's intimacy, due to the type of data that they register.","Therefore it is continued depending on not making a mistake of building, of floor or plants, sector inside the floor or plant, sense of search at the moment to find the parked vehicle, lacking a device that orients and guides him/her to the search vehicle, without it means to add accessories to the vehicle or to have to carry some extra elements in his/her pocket.","The present invention allows to save the mentioned difficulties, using current technology, daily and simple use elements, which hierarchize the areas intended for parking vehicles, for its extra service that offers to the user.","The applicant in the search carried out in the databases linked to the O.E.P.M's web page, it didn't find another system to find parked vehicles, with the characteristics of the one is going to describe after, therefore it isn't known the existence of another similar device."],["OBJECT OF THE INVENTION The present invention which is defended here, consists of a system conformed by several devices that allow to find an automobile or another vehicle parked in areas intended for said purpose, by means of widely used current technology, which are easy to use and do not require installing additional accessories in the vehicle.","For a better service to the user of the parking areas, it is convenient to have a device that orients and guides the user until the parking space of his/her vehicle, who does not have to worry about its location.","BACKGROUND OF THE INVENTION In the current parking, the only existing method to locate a parked vehicle later on, It is the identification of the parking space by means of the letters and digits combination, for example J320.It must be usually completed with the number of the floor or plant and the sector inside the floor or plant, being the localization as follows: floor 3, sector Cinema, street J, parking space 320.The range of colors, is used to delimit the different parking areas.","A form to palliate this problem, is that the user takes note of the code, either memorizing, recording or registering it electronically, or writing it in any element at his/her disposal, what means that he/she will have to transport some extra element in his/her pocket.","In these places any method that is based on the Global Position System (GPS), GLONASS or similar as the next Galilean lacks usefulness, because there are some obstacles (the own building) that interfere in the satellites signs which must be received by the navigator or receiving GPS installed in the vehicle, to define the exact position in coordinates, at the moment he/she leaves it parked.","When the user wants to find his/her parked vehicle, he/she has the same inconvenience with the receiving portable GPS, where previously it was also impossible to register the localization of the parking space in question.","Also this coordinates used to have a short coming of at least 40 meters, unless it arrange differential phase or the DGPS corrections.","In the market there are different optic systems and/or mechanics and/or electronic and/or electromagnetic which solve the problem in question, but their biggest inconvenience reside in the high cost of installation and maintenance, due to its complexity, sophistication and the human personnel who must be devoted to be of that service.","These systems can have interferences of different characteristics which diminish their effectiveness in the supposed solution, and which can transform them in unviable or little practical.","It can consider that some of them attempt against the anonymity and people's intimacy, due to the type of data that they register.","Therefore it is continued depending on not making a mistake of building, of floor or plants, sector inside the floor or plant, sense of search at the moment to find the parked vehicle, lacking a device that orients and guides him/her to the search vehicle, without it means to add accessories to the vehicle or to have to carry some extra elements in his/her pocket.","The present invention allows to save the mentioned difficulties, using current technology, daily and simple use elements, which hierarchize the areas intended for parking vehicles, for its extra service that offers to the user.","The applicant in the search carried out in the databases linked to the O.E.P.M's web page, it didn't find another system to find parked vehicles, with the characteristics of the one is going to describe after, therefore it isn't known the existence of another similar device.","EXPLANATION OF THE INVENTION The main novelty of the invention, object of this presentation, it is based on the registration or storage of the localization of parked vehicle, in an appropriate support and its later process to be able to orient and guide the user until his/her vehicle, without getting wrong.","The cards with magnetic band or intelligent chip, it is the more spread support in this type of establishments, in order to getting paid for the time that the vehicle was parked, therefore it takes advantage of the mentioned support to store the localization, keeping up the same quantity of elements which are used at the moment and avoiding any type of added annoyance.","The system is characterized by its modular design which consists of the following components: a suitably programmed microcontroller, an audiovisual electronic alerting unit, a Smartcard connector, a bar code laser scanner, a LCD screen, a power source connected to the general net of electric fixtures, a presentation box, Smartcard, ticket-card with bar code, computer programs and flat relative cartesian coordinates, everything appropriately combine to get the objective.","The localization of the parked vehicle is defined by means of a system of flat relative cartesian coordinates, own and unique of the building or area intended for such purposed, that is materialized in the code to store.","To this code is added the identification of the parking, to avoid the search in a mistaken building, also adding other data for an easy search.","A previous study of the parking, the later installation project of the invention now defended and the necessary investment, in such way that it were not a cost to the user, it will condition the quantity of each one of mentioned types posts which allow to define the localization code in relation to each parking space of itself, for its using after materializing the invention physically.","It is named “Registering Post”, to the place by means of a device registers or stores the localization code of the parked vehicle, being only used the nearest to the parking space in question, so that the later search were the correct one and “Orienting Post” to the place that by means of another device, partially different to the previous one, it gives the necessary orientation and guided to find the parked vehicle, being able to use whichever and every time that it were necessary until finding the searched place.","Once the vehicle is parked, the “Registering Post” registers or stores the localization code of the parking space, by means of the option that the parking has installed: a) If it is bar code it is registered in the magnetic band card by means of a stamp of automatic ink (simile Trodat), pressing it by hand or also taking away the ticket-corresponding card, from the recipient intended for such purposed.","b) If it is Smartcard, it is inserted in the corresponding connector, to record the data that have to store, taking away the card after finishing the recording, the one is indicated through an audiovisual sign.","The “Registering Post” of bar code, uses the stamp of automatic ink, where the usual support of rubber is stuck, this has recorded the localization code, needing a minimum cleaning maintenance and ink pad replacement with ink, as well as the verification of the quality impression.","It also uses the ticket-card deposited in a recipient that needs the corresponding replacement, which are printed by means of a normal characteristics computer and the use of an appropriate shears, for its cut, generating a hand work.","The printing program ticket-card, has stored the localization codes of each parking space.","The “Registering Post” of Smartcard is electronic and automatic and when detecting the insertion of the card in the Smartcard connector, it carries out the recording of the data belonging to the localization code.","The “Orienting Post” has among its devices, the suitable lector of the support used to register or store the localization code of the parked vehicle, being a completely electronic and automatic process.","If the support is of bar code, it has a laser scanner with a good configurations, for the used format among its constituents, so that it is activated and read automatically before the presence inside its influence area, of the magnetic band card or the ticket-card which have registered the corresponding localization code.","The “Orienting Post” for Smartcard support, has a Smartcard connector among its constituents, turning on mechanics-electronically to the microcontroller when the card is inserted in its interior and being carried out the lecturing in automatic way.","The microcontroller processes the localization code, in order to visualize in the retro-illuminated LCD screen, by means of an alphanumeric text, the information that will allow to orient and guide the user which requests it, until the searching parking space.","The different informations which can be visualized, they allow us to know if the building, floor or plants and sector inside the floor or plant are the correct ones, to be able to begin the properly search, when we are in the nearest area to the searched vehicle.","In the system of flat relative cartesian coordinates, the difference in the axis “z” (vertical), will indicate the quantity of floors or plants, expressed in units which it must be up or down, to arrive at the right place.","In most of the establishments intended for parking, the circulation streets inside the same have a distribution of rectangular form, having as norm the axis “x” parallel to the biggest length streets and the axis “y” parallel to the smallest length streets.","When we are in the nearest area to the searched vehicle, the information that is visualized it is the following one: a) steps to walk along the street of the “Orienting Post” (axis “x”), in the indicated direction in the LCD screen and that it coincides with the distance expressed in meters (difference of coordinates axis “x”) existing between the “Orienting Post” and the normal point to the parking space in question, on the street that we are going through.","b) to get to the normal point we must turn to the left or to the right, according to the result of the difference of the coordinates in the axis “y” between the mentioned normal point and the parking space in question c) steps to walk along the normal to the street of the “Orienting Post” (axis “y”), until get to the parked vehicle which was looked for, and that it coincides with the distance expressed in meters (difference of coordinates axis “y”) existing between the mentioned normal point and the parked vehicle.","The invention which is defended here, it checks the different processes, through different computer programs designed especially to such effect and which are registered in the microcontroller memory, during the installation of the system.","The computer program installed in the microcontroller, it is mainly designed to control the following processes: a) the manage of the system of flat relative Cartesian coordinate, in all the calculations that the invention carries out b) the recording and lecturing of the localization code of the parked vehicle in the Smartcard c) the data admission sent by the laser scanner, corresponding to the lecturing of the cards which register the localization code, by means of the corresponding bar code d) the visualization of the different informations in the LCD screen that they orient and guide to the user who requests it, to find his/her vehicle e) the control of audiovisual electronic alerting unit f) reset the system g) the lecturing and memorization of the “Post” coordinates h) the auto-checking of the “Post” correct initiation i) the control of the correct operation of the different devices of the system j) the control of the language change that will be used in the visualization of the informations.","In the “Orienting Post” in order to achieve a biggest universality, it is able to choose the language in which we want to visualized the informations in the LCD screen, being able to achieve this selection, pressing once the idiom button corresponding to the wanted language.","For not increasing the installation costs, a space problem for the buttons and memory to have kept the text to visualize, it limits that selection to less than ten options that will allow to select the more used languages in the area where the system is installed.","The audiovisual electronic alerting unit has the function of indicating in which situation it is, the recording or the lecturing of the localization code of the parked vehicle.","There is a colour visual sign and quantity of Led and an acoustic sign taken place by the loudspeaker that identifies the moment in which the microcontroller is carrying out the lecturing or recording of the mentioned data, also allowing to know easily when these processes finish by means of the different characteristics of the audiovisual sign.","The power sources that the computers use, they improve the modular design and mainly satisfy the necessities of electric power supply to the tension and strength necessary, of the different electronic constituents of the system and being connected to the domiciliary general net of electric fixtures, through their transformers.","The box or wrapping has as main function, the protection of the devices that contains inside, the execution of the electric security norms and the perfect visualization of the “Posts”, for their identification, also taking advantage of its construction in order to facilitate the system assembly, to show the logo of the product, the use guide itself, a commercial presentation of the product and others.","DESCRIPTION OF THE DRAWINGS To facilitate the understanding of the present invention characteristics, it is added below a descriptive memory of several figures, in those that with illustrative and never limitative character, the following thing has been represented: The FIG.","1 shows a perspective view of a small parking area, where we distinguish four parking space and two columns belonging to the building that have anchored the “Registering Post” (1.1) and the “Orienting Post” (1.2).","The FIG.","2 shows a perspective view of the two types of available data support, the Smartcard (2.1) and the magnetic band card or the ticket-card, with its bar code (2.2) in the appropriate format.","The FIG.","3 shows a perspective view, of a part of the two types of available “Registering Post, the Smartcard (3.1) and the bar code (3.2), with its stamp of automatic ink and recipient.","The FIG.","4 shows a perspective view of the two types of available “Orienting Post”, the Smartcard (4.1) and the bar code (4.2) The FIG.","5 shows a perspective view of a small parking area, where we distinguish the way for finding the vehicle parked starting from the information facilitated by the “Orienting Post” (1.2) and according to the system of flat relative cartesian coordinates.","The FIG.","6 shows a perspective view of the LCD screen (6) visualizing some of the informations to orient and guide the user that requests it and the buttons (6.1) that allow to change the language of the text to read.","EXPOSITION OF THE WAY TO CARRYOUT THE INVENTION According to the previously enunciated, the present invention refers to the Systems for Finding Vehicles Parked Inside Buildings and Roofed Surfaces, characterized by a modular design which consists of the following modules: a suitably programmed microcontroller, an audiovisual electronic alerting unit (7), a Smartcard connector (5), a barcode laser scanner (8), a retro-illuminated LCD screen (6), a power source connected to the general net of electric fixtures, a presentation box, the Smartcard (2.1) and the magnetic band card or the ticket-card with bar code (2.2).","The Smartcard (2.1) has normal characteristics, not needing protection neither special security, should allow the use of the protocol I2C during the lecturing and recording what places, it in the range of daily use.","The Smartcard connector (5) for intelligent card (2.1) has normal characteristics, doing the contact by touching down with the objective of not damage the card contact existing a detector contact which indicates the insertion itself.","When the lecturing or recording of the card finishes, the system is prepared for the different expulsion types of itself that are automatic or manual, being able to use in the realization of this invention, the simple method of pushing and dropping.","The audiovisual electronic alerting unit (7) is compound of a led group (7.1) and a buzzer (7.2), generating a visual sign (quantity and colour) and another acoustic, by means of which it is identified the processes that the microcontroller is carrying out.","The laser scanner (8) for the lecturing of the barcode ticket (2.2), has normal characteristics, configured for the automatic lecturing when the ticket is in the influence scanner area.","The system is prepared for different types of format codes and inter phase of communications, being able to use the RS-232 in the data transmission with the microcontroller.","The retro-illuminated LCD screen (6), alphanumeric has normal characteristics, it optimizes the visualization of the informations that are shown to the user that requests it.","Among them there is screens of 80 characters (20×4) distributed in 4 lines, with big characters that collaborate with the clarity of what is written.","The communication with the microcontroller is carried out through a bus of data and another of commands, being allowed its control in that way.","The buttons (6.1) that allow to carry out the language selection in which the informations will be visualized in the LCD screen, they are completely normal and they don't have any special characteristics to keep in mind.","The power source is the necessary to be able to give the tension of 5 volt to the electronic constituents of the system, being linked to the domiciliary general net of electric fixtures.","The biggest consumption is due to the retro illumination of the LCD screen that is about 1000 mA that must be added to the 300 mA that the rest of the electronic circuit can consume.","The microcontroller has normal characteristics, being without inconveniences in the market, being of 18, 28 or 40 the quantity paws which are used for its communication with the electronic circuit that surrounds it.","The speed is not of importance, due to the processes are not complex neither long, therefore there are not delays in the times of execution.","The quantity of necessary memory intended for programs and data, it is satisfied using microcontroller families of more than 1 Kbytes for the first one and of 256 bytes for the second one.","The box or wrapping allows to set up, to hold and to protect the different constituents, being its perfect visualization the one makes easy the identification of the “Posts.” They take the manual use of the system and they are anchored by means of screws to the walls or columns, where they are installed according to the optimum of the system.","In the reproduction of this invention for an expert, the existence of variations in materials, forms, size and constituents disposition, these don't affect to the essential of itself, which were described in a no limitative way."]],"string":"[\n [\n \"System for finding vehicles parked inside buildings and roofed surfaces\",\n \"To find an automobile or another vehicle that it was left parked, in spaces intended for such purpose, by means of widely used current technology, of simple use and without adding accessories to the vehicle.\",\n \"Several devices, characterized by their modular design, the flat relative cartesian coordinates and computer programs, constitute the system jointly, that orienting and guiding the user to the parking space of his/her vehicle, who does not have to worry about its location.\",\n \"This invention uses the following constituents: presentation box, programmed microcontroller, audiovisual electronic alerting unit (7), Smartcard connector (5), barcode laser scanner (8), LCD screen (6), power source, Smartcard (2.1), ticket-card with bar code (2.2), coordinates and computer programs.\",\n \"The “Registering Post” (1.1) defines and registers the localization of the parked vehicle and the “Orienting Post” (1.2) visualizes the information in its LCD screen (6) to orient and guide the user, until finding his/her parked vehicle.\"\n ],\n [\n \"1.Systems for Finding Vehicles Parked Inside Buildings and Roofed Surfaces, characterized by having got two types of modules (1), that can contain a Smartcard connector, a bar code laser scanner, a retro-illuminated LCD screen, a languages selector, an audiovisual electronic alerting unit, a power source connected to the general net of electric fixtures, a microcontroller integrated in the electronic circuit and a presentation box.\",\n \"2.The system according to the preceding claim, characterized by having got two types of modules (1): a) the “Registering Post” (1.1) allows to register or store in the data support (2) used by the parking, the localization code of the parked vehicle, being necessary its installation only in parking areas.\",\n \"b) the “Orienting Post” (1.2) allows to orient and guide the user until finding his/her parked vehicle, based on the lecturing registration of the data support (2), being able to install in accesses, entries, corridors, stairways, parking areas, and so on, where the quantity of these modules will be condicionated by the optimum service.\",\n \"3.The system according to the preceding claims, characterized because the “Registering Post” (3) has got to use two types of data support (2) in the localization register of the parked vehicle: a) the Smartcard (2.1), that is electronically recording, when it is inserted in the connector (5).\",\n \"b) ticket-card of bar code (2.2), that must be remove from recipient (3.2) puts for such effect.\",\n \"c) magnetic band card where the bar code (2.2) is registered, by means of the stamp of automatic ink (simile Trodat), pressing it by hand.\",\n \"(3.2).\",\n \"4.The system according to the preceding claims, characterized because the “Orienting Post” (4) has got to recover the localization of the parked vehicle, registered from the data support (2), of two types of lectors: a) Smartcard connector (5) being activated the reading, when the Smartcard (2.1) is inserted.\",\n \"b) laser scanner (8) being activated the lecturing, when the magnetic band card (2.2) passes by its influence area, or the ticket-card (2.2) where the bar code was registered, according to the used format.\",\n \"5.The system according to the preceding claims, characterized because the “Post Orienting” (4) has got a retro-illuminated LCD screen (6), with an alphanumeric text to visualize the information that allows to orient and guide the user that requests it, until finding his/her parked vehicle.\",\n \"6.The system according to the preceding claims, characterized because the two types of modules (1), to identify what type of process is executing by the microcontroller, they have got the audiovisual electronic alerting unit (7), which is compounded by a visual colour sign and quantity of Led (7.1) and an acoustic sign taken place by the loudspeaker (7.2).\",\n \"7.The system according to the preceding claims, characterized because the two types of modules (1), have got a microcontroller integrated in an electronic circuit, which controls all the processes that are executed in the system, by means of a structured computer program and designed especially to develop the necessary functions and that it is registered in the microcontroller memory, when the system is installed.\",\n \"8.The system according to the preceding claims, characterized because the computer program that is installed in the microcontroller, it mainly controls the following processes: a) the manage system of flat relative cartesian coordinates.\",\n \"b) the recording and lecturing in the Smartcard of the localization code of the parked vehicle.\",\n \"c) the data admission sent by the laser scanner, corresponding to the lecturing of cards with bar code d) the visualization in the LCD screen of the different informations.\",\n \"e) the control of the audiovisual electronic alerting unit f) reset the system g) the lecturing and memorization of the “Post” coordinates h) the auto-checking of the “Post” correct initiation i) the control of the different devices correct operations of the system j) the control of the language change in which the informations are visualized.\",\n \"9.The system according to the preceding claims, characterized by using a system of flat relative cartesian coordinates, to define the localization of each parked vehicle, which they are own and unique of each parking space intended for parking inside the same building or space used for parking.\",\n \"10.The system according to the preceding claims, characterized because in the system of flat relative cartesian coordinates, the difference in the axis “z” indicates the quantity of floors or plants expressed in unities which it must be up or down, to get to the right place, the difference in the axis “x” and in the sense that the LCD screen indicates, it expresses the quantity of meters the parked vehicle is, according to the walking by the street of the “Orienting Post” and the difference in the axis “y” expresses the quantity of meters the parked vehicle is far from the previous street to the right or to the left, as it is the difference in the axis “y” positive or negative.\",\n \"11.The system according to the preceding claims, characterized because the Smartcard (2.1), used as data support (2), it isn't personalized to a parking in exclusiveness, being able to use the same card, in all the parkings or parking that have installed the system of this invention.\",\n \"12.The system according to the preceding claims, characterized because the “Orienting Post” possesses a series of buttons that allow to choose the language in which we want to visualize the informations in the LCD screen, pressing once the corresponding to the selected language.\"\n ],\n [\n \"<SOH> BACKGROUND OF THE INVENTION <EOH>In the current parking, the only existing method to locate a parked vehicle later on, It is the identification of the parking space by means of the letters and digits combination, for example J 320 .\",\n \"It must be usually completed with the number of the floor or plant and the sector inside the floor or plant, being the localization as follows: floor 3 , sector Cinema, street J, parking space 320 .\",\n \"The range of colors, is used to delimit the different parking areas.\",\n \"A form to palliate this problem, is that the user takes note of the code, either memorizing, recording or registering it electronically, or writing it in any element at his/her disposal, what means that he/she will have to transport some extra element in his/her pocket.\",\n \"In these places any method that is based on the Global Position System (GPS), GLONASS or similar as the next Galilean lacks usefulness, because there are some obstacles (the own building) that interfere in the satellites signs which must be received by the navigator or receiving GPS installed in the vehicle, to define the exact position in coordinates, at the moment he/she leaves it parked.\",\n \"When the user wants to find his/her parked vehicle, he/she has the same inconvenience with the receiving portable GPS, where previously it was also impossible to register the localization of the parking space in question.\",\n \"Also this coordinates used to have a short coming of at least 40 meters, unless it arrange differential phase or the DGPS corrections.\",\n \"In the market there are different optic systems and/or mechanics and/or electronic and/or electromagnetic which solve the problem in question, but their biggest inconvenience reside in the high cost of installation and maintenance, due to its complexity, sophistication and the human personnel who must be devoted to be of that service.\",\n \"These systems can have interferences of different characteristics which diminish their effectiveness in the supposed solution, and which can transform them in unviable or little practical.\",\n \"It can consider that some of them attempt against the anonymity and people's intimacy, due to the type of data that they register.\",\n \"Therefore it is continued depending on not making a mistake of building, of floor or plants, sector inside the floor or plant, sense of search at the moment to find the parked vehicle, lacking a device that orients and guides him/her to the search vehicle, without it means to add accessories to the vehicle or to have to carry some extra elements in his/her pocket.\",\n \"The present invention allows to save the mentioned difficulties, using current technology, daily and simple use elements, which hierarchize the areas intended for parking vehicles, for its extra service that offers to the user.\",\n \"The applicant in the search carried out in the databases linked to the O.E.P.M's web page, it didn't find another system to find parked vehicles, with the characteristics of the one is going to describe after, therefore it isn't known the existence of another similar device.\"\n ],\n [\n \"OBJECT OF THE INVENTION The present invention which is defended here, consists of a system conformed by several devices that allow to find an automobile or another vehicle parked in areas intended for said purpose, by means of widely used current technology, which are easy to use and do not require installing additional accessories in the vehicle.\",\n \"For a better service to the user of the parking areas, it is convenient to have a device that orients and guides the user until the parking space of his/her vehicle, who does not have to worry about its location.\",\n \"BACKGROUND OF THE INVENTION In the current parking, the only existing method to locate a parked vehicle later on, It is the identification of the parking space by means of the letters and digits combination, for example J320.It must be usually completed with the number of the floor or plant and the sector inside the floor or plant, being the localization as follows: floor 3, sector Cinema, street J, parking space 320.The range of colors, is used to delimit the different parking areas.\",\n \"A form to palliate this problem, is that the user takes note of the code, either memorizing, recording or registering it electronically, or writing it in any element at his/her disposal, what means that he/she will have to transport some extra element in his/her pocket.\",\n \"In these places any method that is based on the Global Position System (GPS), GLONASS or similar as the next Galilean lacks usefulness, because there are some obstacles (the own building) that interfere in the satellites signs which must be received by the navigator or receiving GPS installed in the vehicle, to define the exact position in coordinates, at the moment he/she leaves it parked.\",\n \"When the user wants to find his/her parked vehicle, he/she has the same inconvenience with the receiving portable GPS, where previously it was also impossible to register the localization of the parking space in question.\",\n \"Also this coordinates used to have a short coming of at least 40 meters, unless it arrange differential phase or the DGPS corrections.\",\n \"In the market there are different optic systems and/or mechanics and/or electronic and/or electromagnetic which solve the problem in question, but their biggest inconvenience reside in the high cost of installation and maintenance, due to its complexity, sophistication and the human personnel who must be devoted to be of that service.\",\n \"These systems can have interferences of different characteristics which diminish their effectiveness in the supposed solution, and which can transform them in unviable or little practical.\",\n \"It can consider that some of them attempt against the anonymity and people's intimacy, due to the type of data that they register.\",\n \"Therefore it is continued depending on not making a mistake of building, of floor or plants, sector inside the floor or plant, sense of search at the moment to find the parked vehicle, lacking a device that orients and guides him/her to the search vehicle, without it means to add accessories to the vehicle or to have to carry some extra elements in his/her pocket.\",\n \"The present invention allows to save the mentioned difficulties, using current technology, daily and simple use elements, which hierarchize the areas intended for parking vehicles, for its extra service that offers to the user.\",\n \"The applicant in the search carried out in the databases linked to the O.E.P.M's web page, it didn't find another system to find parked vehicles, with the characteristics of the one is going to describe after, therefore it isn't known the existence of another similar device.\",\n \"EXPLANATION OF THE INVENTION The main novelty of the invention, object of this presentation, it is based on the registration or storage of the localization of parked vehicle, in an appropriate support and its later process to be able to orient and guide the user until his/her vehicle, without getting wrong.\",\n \"The cards with magnetic band or intelligent chip, it is the more spread support in this type of establishments, in order to getting paid for the time that the vehicle was parked, therefore it takes advantage of the mentioned support to store the localization, keeping up the same quantity of elements which are used at the moment and avoiding any type of added annoyance.\",\n \"The system is characterized by its modular design which consists of the following components: a suitably programmed microcontroller, an audiovisual electronic alerting unit, a Smartcard connector, a bar code laser scanner, a LCD screen, a power source connected to the general net of electric fixtures, a presentation box, Smartcard, ticket-card with bar code, computer programs and flat relative cartesian coordinates, everything appropriately combine to get the objective.\",\n \"The localization of the parked vehicle is defined by means of a system of flat relative cartesian coordinates, own and unique of the building or area intended for such purposed, that is materialized in the code to store.\",\n \"To this code is added the identification of the parking, to avoid the search in a mistaken building, also adding other data for an easy search.\",\n \"A previous study of the parking, the later installation project of the invention now defended and the necessary investment, in such way that it were not a cost to the user, it will condition the quantity of each one of mentioned types posts which allow to define the localization code in relation to each parking space of itself, for its using after materializing the invention physically.\",\n \"It is named “Registering Post”, to the place by means of a device registers or stores the localization code of the parked vehicle, being only used the nearest to the parking space in question, so that the later search were the correct one and “Orienting Post” to the place that by means of another device, partially different to the previous one, it gives the necessary orientation and guided to find the parked vehicle, being able to use whichever and every time that it were necessary until finding the searched place.\",\n \"Once the vehicle is parked, the “Registering Post” registers or stores the localization code of the parking space, by means of the option that the parking has installed: a) If it is bar code it is registered in the magnetic band card by means of a stamp of automatic ink (simile Trodat), pressing it by hand or also taking away the ticket-corresponding card, from the recipient intended for such purposed.\",\n \"b) If it is Smartcard, it is inserted in the corresponding connector, to record the data that have to store, taking away the card after finishing the recording, the one is indicated through an audiovisual sign.\",\n \"The “Registering Post” of bar code, uses the stamp of automatic ink, where the usual support of rubber is stuck, this has recorded the localization code, needing a minimum cleaning maintenance and ink pad replacement with ink, as well as the verification of the quality impression.\",\n \"It also uses the ticket-card deposited in a recipient that needs the corresponding replacement, which are printed by means of a normal characteristics computer and the use of an appropriate shears, for its cut, generating a hand work.\",\n \"The printing program ticket-card, has stored the localization codes of each parking space.\",\n \"The “Registering Post” of Smartcard is electronic and automatic and when detecting the insertion of the card in the Smartcard connector, it carries out the recording of the data belonging to the localization code.\",\n \"The “Orienting Post” has among its devices, the suitable lector of the support used to register or store the localization code of the parked vehicle, being a completely electronic and automatic process.\",\n \"If the support is of bar code, it has a laser scanner with a good configurations, for the used format among its constituents, so that it is activated and read automatically before the presence inside its influence area, of the magnetic band card or the ticket-card which have registered the corresponding localization code.\",\n \"The “Orienting Post” for Smartcard support, has a Smartcard connector among its constituents, turning on mechanics-electronically to the microcontroller when the card is inserted in its interior and being carried out the lecturing in automatic way.\",\n \"The microcontroller processes the localization code, in order to visualize in the retro-illuminated LCD screen, by means of an alphanumeric text, the information that will allow to orient and guide the user which requests it, until the searching parking space.\",\n \"The different informations which can be visualized, they allow us to know if the building, floor or plants and sector inside the floor or plant are the correct ones, to be able to begin the properly search, when we are in the nearest area to the searched vehicle.\",\n \"In the system of flat relative cartesian coordinates, the difference in the axis “z” (vertical), will indicate the quantity of floors or plants, expressed in units which it must be up or down, to arrive at the right place.\",\n \"In most of the establishments intended for parking, the circulation streets inside the same have a distribution of rectangular form, having as norm the axis “x” parallel to the biggest length streets and the axis “y” parallel to the smallest length streets.\",\n \"When we are in the nearest area to the searched vehicle, the information that is visualized it is the following one: a) steps to walk along the street of the “Orienting Post” (axis “x”), in the indicated direction in the LCD screen and that it coincides with the distance expressed in meters (difference of coordinates axis “x”) existing between the “Orienting Post” and the normal point to the parking space in question, on the street that we are going through.\",\n \"b) to get to the normal point we must turn to the left or to the right, according to the result of the difference of the coordinates in the axis “y” between the mentioned normal point and the parking space in question c) steps to walk along the normal to the street of the “Orienting Post” (axis “y”), until get to the parked vehicle which was looked for, and that it coincides with the distance expressed in meters (difference of coordinates axis “y”) existing between the mentioned normal point and the parked vehicle.\",\n \"The invention which is defended here, it checks the different processes, through different computer programs designed especially to such effect and which are registered in the microcontroller memory, during the installation of the system.\",\n \"The computer program installed in the microcontroller, it is mainly designed to control the following processes: a) the manage of the system of flat relative Cartesian coordinate, in all the calculations that the invention carries out b) the recording and lecturing of the localization code of the parked vehicle in the Smartcard c) the data admission sent by the laser scanner, corresponding to the lecturing of the cards which register the localization code, by means of the corresponding bar code d) the visualization of the different informations in the LCD screen that they orient and guide to the user who requests it, to find his/her vehicle e) the control of audiovisual electronic alerting unit f) reset the system g) the lecturing and memorization of the “Post” coordinates h) the auto-checking of the “Post” correct initiation i) the control of the correct operation of the different devices of the system j) the control of the language change that will be used in the visualization of the informations.\",\n \"In the “Orienting Post” in order to achieve a biggest universality, it is able to choose the language in which we want to visualized the informations in the LCD screen, being able to achieve this selection, pressing once the idiom button corresponding to the wanted language.\",\n \"For not increasing the installation costs, a space problem for the buttons and memory to have kept the text to visualize, it limits that selection to less than ten options that will allow to select the more used languages in the area where the system is installed.\",\n \"The audiovisual electronic alerting unit has the function of indicating in which situation it is, the recording or the lecturing of the localization code of the parked vehicle.\",\n \"There is a colour visual sign and quantity of Led and an acoustic sign taken place by the loudspeaker that identifies the moment in which the microcontroller is carrying out the lecturing or recording of the mentioned data, also allowing to know easily when these processes finish by means of the different characteristics of the audiovisual sign.\",\n \"The power sources that the computers use, they improve the modular design and mainly satisfy the necessities of electric power supply to the tension and strength necessary, of the different electronic constituents of the system and being connected to the domiciliary general net of electric fixtures, through their transformers.\",\n \"The box or wrapping has as main function, the protection of the devices that contains inside, the execution of the electric security norms and the perfect visualization of the “Posts”, for their identification, also taking advantage of its construction in order to facilitate the system assembly, to show the logo of the product, the use guide itself, a commercial presentation of the product and others.\",\n \"DESCRIPTION OF THE DRAWINGS To facilitate the understanding of the present invention characteristics, it is added below a descriptive memory of several figures, in those that with illustrative and never limitative character, the following thing has been represented: The FIG.\",\n \"1 shows a perspective view of a small parking area, where we distinguish four parking space and two columns belonging to the building that have anchored the “Registering Post” (1.1) and the “Orienting Post” (1.2).\",\n \"The FIG.\",\n \"2 shows a perspective view of the two types of available data support, the Smartcard (2.1) and the magnetic band card or the ticket-card, with its bar code (2.2) in the appropriate format.\",\n \"The FIG.\",\n \"3 shows a perspective view, of a part of the two types of available “Registering Post, the Smartcard (3.1) and the bar code (3.2), with its stamp of automatic ink and recipient.\",\n \"The FIG.\",\n \"4 shows a perspective view of the two types of available “Orienting Post”, the Smartcard (4.1) and the bar code (4.2) The FIG.\",\n \"5 shows a perspective view of a small parking area, where we distinguish the way for finding the vehicle parked starting from the information facilitated by the “Orienting Post” (1.2) and according to the system of flat relative cartesian coordinates.\",\n \"The FIG.\",\n \"6 shows a perspective view of the LCD screen (6) visualizing some of the informations to orient and guide the user that requests it and the buttons (6.1) that allow to change the language of the text to read.\",\n \"EXPOSITION OF THE WAY TO CARRYOUT THE INVENTION According to the previously enunciated, the present invention refers to the Systems for Finding Vehicles Parked Inside Buildings and Roofed Surfaces, characterized by a modular design which consists of the following modules: a suitably programmed microcontroller, an audiovisual electronic alerting unit (7), a Smartcard connector (5), a barcode laser scanner (8), a retro-illuminated LCD screen (6), a power source connected to the general net of electric fixtures, a presentation box, the Smartcard (2.1) and the magnetic band card or the ticket-card with bar code (2.2).\",\n \"The Smartcard (2.1) has normal characteristics, not needing protection neither special security, should allow the use of the protocol I2C during the lecturing and recording what places, it in the range of daily use.\",\n \"The Smartcard connector (5) for intelligent card (2.1) has normal characteristics, doing the contact by touching down with the objective of not damage the card contact existing a detector contact which indicates the insertion itself.\",\n \"When the lecturing or recording of the card finishes, the system is prepared for the different expulsion types of itself that are automatic or manual, being able to use in the realization of this invention, the simple method of pushing and dropping.\",\n \"The audiovisual electronic alerting unit (7) is compound of a led group (7.1) and a buzzer (7.2), generating a visual sign (quantity and colour) and another acoustic, by means of which it is identified the processes that the microcontroller is carrying out.\",\n \"The laser scanner (8) for the lecturing of the barcode ticket (2.2), has normal characteristics, configured for the automatic lecturing when the ticket is in the influence scanner area.\",\n \"The system is prepared for different types of format codes and inter phase of communications, being able to use the RS-232 in the data transmission with the microcontroller.\",\n \"The retro-illuminated LCD screen (6), alphanumeric has normal characteristics, it optimizes the visualization of the informations that are shown to the user that requests it.\",\n \"Among them there is screens of 80 characters (20×4) distributed in 4 lines, with big characters that collaborate with the clarity of what is written.\",\n \"The communication with the microcontroller is carried out through a bus of data and another of commands, being allowed its control in that way.\",\n \"The buttons (6.1) that allow to carry out the language selection in which the informations will be visualized in the LCD screen, they are completely normal and they don't have any special characteristics to keep in mind.\",\n \"The power source is the necessary to be able to give the tension of 5 volt to the electronic constituents of the system, being linked to the domiciliary general net of electric fixtures.\",\n \"The biggest consumption is due to the retro illumination of the LCD screen that is about 1000 mA that must be added to the 300 mA that the rest of the electronic circuit can consume.\",\n \"The microcontroller has normal characteristics, being without inconveniences in the market, being of 18, 28 or 40 the quantity paws which are used for its communication with the electronic circuit that surrounds it.\",\n \"The speed is not of importance, due to the processes are not complex neither long, therefore there are not delays in the times of execution.\",\n \"The quantity of necessary memory intended for programs and data, it is satisfied using microcontroller families of more than 1 Kbytes for the first one and of 256 bytes for the second one.\",\n \"The box or wrapping allows to set up, to hold and to protect the different constituents, being its perfect visualization the one makes easy the identification of the “Posts.” They take the manual use of the system and they are anchored by means of screws to the walls or columns, where they are installed according to the optimum of the system.\",\n \"In the reproduction of this invention for an expert, the existence of variations in materials, forms, size and constituents disposition, these don't affect to the essential of itself, which were described in a no limitative way.\"\n 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"Type II thioesterase from streptomyces coelicolor a3(2) and the coding sequence",
"The invention concerns a biosynthesis of nonaramatic polyketide compounds.",
"More particularly, the present invention relates to a isolated polynucleotide molecule, a nucleic acid vector comprising the molecule, and i... | Patent_10473092 |
[
[
"Process for producing trunkamide a compounds",
"Trunkamide A and other cyloheptatides can be made by solid phase synthesis of a linear precursor."
],
[
"1 The preparation of a cycloheptapeptide by a solid phase synthesis of a linear heptapeptide precursor.",
"2.A process for preparing a cyclo... | Patent_10473096 |
[
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"Compounds and methods",
"Compounds of this invention are non-peptide, reversible inhibitors of type 2 methionine aminopeptidase, useful in treating conditions mediated by angiogenesis, such as cancer, haemangioma, proliferative retinopathy, rheumatoid arthritis, atherosclerotic neovascularization, psoria... | Patent_10473106 |
[
[
"Record medium with different latencies",
"Latency information is used for coding of additional information in a record medium.",
"This additional information which gets embossed in the record medium might be used for verification purposes in that only storage media with the correct latency informatio... | Patent_10473112 |
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"Valve",
"A valve for an aerosol comprises a valve body 1 and a surrounding rubber sleeve 2.",
"The body defines an inlet and outlet passages 3, 4 and bores 5, 6 connecting these passages to the surface of the valve.",
"When the pressure of fluid in the inlet passage 3 and bore 5 builds to a suffi... | Patent_10473118 |
[
[
"Printing couple in a printing machine with a pivotable transfer cylinder",
"A printing couple in a printing machine is comprised of at least three cylinders, a forme cylinder, a transfer cylinder and a counter-pressure cylinder.",
"The counter-pressure cylinder forms a printing location or point in c... | Patent_10473148 |
[
[
"Microarray gene expression profiling in clear cell renal cell carcinoma : prognosis and drug target identification",
"A nucleic acid probe or a novel set of such probes in a microarray is provided.",
"The probe or probe set is useful in the prognosis of patients with clear cell renal cell carcinoma (... | Patent_10473173 |
[
[
"Tyre for vehicle wheels with a reinforced bead",
"Tyre for vehicle wheels comprising; a) a carcass structure having at least one carcass ply associated with respective left and right bead wires, each bead wire being enclosed in a respective bead, said bead comprising a bead filler b) a belt structure com... | Patent_10473174 |
[
[
"Methods of delivery of exogenous proteins to the cytosol and uses thereof",
"The present invention is directed to a method for delivering exogenous proteins to the cytosol, by binding a target antigen (such as a protein) to a transport factor that contains a fragment of a bipartite protein exotoxin, but ... | Patent_10473190 |
[
[
"Leinamycin biosynthesis gene cluster and its components and their uses",
"This invention provides detailed sequence analysis and characterization of the gene cluster responsible for the synthesis of leinamycin in Streptomyces atroolivaceus.",
"The leinamycin gene cluster provides a hybrid polyketide ... | Patent_10473193 |
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[
"Catalyst composition",
"A catalyst composition for the oxidation of carbon monoxide and volatile organic compounds and for hydrogenation reactions comprises at least two different high surface area oxide support materials wherein at least one of the high surface area support material supports at least on... | Patent_10473209 |
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"Gene regulation II",
"We describe a transgenic non-human animal comprising a heterologous nucleic acid binding polypeptide which binds to a target gene and modulates its expression, in which the heterologous nucleic acid binding polypeptide is encoded by a transgene, and in which the expression of a targ... | Patent_10473238 |
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[
"Alpha conotoxin peptides with analgesic properties",
"This invention relates to novel α-conotoxin-like peptides comprising the following sequence of amino acids: Xaa1CCSXaa2Xaa3Xaa4CXaa5Xaa6Xaa7Xaa8Xaa9Xaa10Xaa11C—NH2 in which Xaa1 is G or D; Xaa3 is proline, hydroxyproline or glutamine; each of Xaa2 to ... | Patent_10473246 |
[
[
"Method of Detecting the Expression of Aspergillus Gene",
"The present invention provides a method of detecting the expression of a koji mold gene, and a DNA for specifically assaying the fermentation conditions for a koji mold.",
"Specifically, the DNA for specifically assaying the fermentation condi... | Patent_10473247 |
[
[
"External skin compositions",
"The external skin care composition of the present invention comprises N-acetylglucosamine and at least one member selected from the group consisting of retinoid and pro-vitamin A.",
"The external skin care composition of the present invention has an effect of promoting t... | Patent_10473293 |
[
[
"Tread pattern for car tire",
"A tyre for a motor car includes a tread and two shoulders.",
"The tread includes at least one circumferential row of blocks disposed between first and second circumferential grooves.",
"Each of the blocks is delimited by a section of the first circumferential groove ... | Patent_10473304 |
[
[
"Microchip pileup type chemical reaction system",
"A microchip pileup type chemical reaction system characterized in that a specified number of microchips, each having a reaction material liquid introducing section, a reaction product liquid discharge section and a reaction region, i.e.",
"microchanne... | Patent_10473336 |
[
[
"Isolated human transporter proteins, nucleic acid molecules encoding human transporter proteins, and uses thereof",
"The present invention provides amino acid sequences of peptides that are encoded by genes within the human genome, the transporter peptides of the present invention.",
"The present inv... | Patent_10473338 |
[
[
"Isolated human ras- like proteins, nucleic acid molecules encoding these human ras-like proteins , and uses thereof",
"The present invention provides amino acid sequences of polypeptides that are encoded by genes within the human genome, the Ras-like protein polypeptides of the present invention.",
"... | Patent_10473340 |
[
[
"System and method for management of remote devices in a network",
"A system and method of managing devices connected to a network by restricted bandwidth links, where a restricted bandwidth link comprises a wireless or other link which has a relatively limited amount of bandwidth available and/or a link ... | Patent_10473342 |
[
[
"Cartilaginous neo-tissue capable of being grafted",
"A cartilaginous neo-tissue that is capable of being grafted is constituted by rows of approximately parallel cells with a cell maturation gradient orientated from a predetermined zone towards its periphery.",
"During preparation of the cartilaginou... | Patent_10473359 |
[
[
"Materials, methods, and uses for photochemical generation of acids and/or radical species",
"The present invention provides compounds and compositions, which include: at least one chromophore having strong simultaneous two-photon or multi-photon absorptivity; at least one acid- or radical-generator in cl... | Patent_10473365 |
[
[
"Automatic number plate recognition system",
"A handheld automatic number plate recognition system comprising a handheld personal computer and a camera, the camera being operable to capture images and the computer including a processor to identify a number plate from an image captured by the camera."
],... | Patent_10473381 |
[
[
"Modification of organelle metabolism by unc-51-like kinases roma1 or 2tm proteins",
"The invention discloses polypeptides (Unc-51 kinase, ROMA1, and/or 2TM protein) affecting the activity of Uncoupling Proteins (UCPs), thereby leading to an altered mitochondrial activity and thus contributing to membrane... | Patent_10473404 |
[
[
"Viral vectors",
"A Human Immunodeficiency Virus (HIV-2) vector having a mutation within a packaging signal such that viral RNA is not packaged within an HIV-2 capsid is described.",
"A further vector comprises an HIV-2 packaging signal and a heterologous gene capable of being expressed in the vector.... | Patent_10473412 |
[
[
"Demodulator and receiver using same",
"A high performance demodulator able to realize a further wide band property, low distortion characteristics, and low power consumption in comparison with a conventional multi-port demodulator and having a small fluctuation in characteristics with respect to temperat... | Patent_10473417 |
[
[
"Protein modification and maintenance molecules",
"The invention provides human protein modification and maintenance molecules (PMOD) and polynucleotides which identify and encode PMOD.",
"The invention also provides expression vectors, host cells, antibodies, agonists, and antagonists.",
"The inv... | Patent_10473451 |
[
[
"Peptides and antibodies to muc 1 proteins",
"The invention relates to methods of inhibiting proliferation or growth of tumor cells and/or inducing cell death relates to the use of antibodies, hybridomas and pharmaceutical compositions containing same, for inhibiting growth or proliferation and inducing d... | Patent_10473484 |
[
[
"Method of making frozen products with a characteristic form from at least one flowable type of material, apparatus and use hereof",
"The invention relates to a method of making frozen products with a characteristic form from at least one flowable type of material, the frozen products including edible ice... | Patent_10473506 |
[
[
"Site- targeted transformation using amplification vectors",
"A process of causing a targeted integration of DNA of interest into a plynt cell nuclear genome, comprising; i) providing plant cells with an amplification vector, or a precursor thereof, capable of autonomous replication in plant cells, said v... | Patent_10473508 |
[
[
"Secreted proteins",
"The invention provides human secreted proteins (SECP) and polynucleotides which identify and encode SECP.",
"The invention also provides expression vectors, host cells, antibodies, agonists, and antagonists.",
"The invention also provides methods for diagnosing, treating, or ... | Patent_10473519 |
[
[
"Gastrokines and derived peptides including inhibitors",
"A novel group of gastrokines called Gastric Antrum Mucosal Protein is characterized.",
"A member of the group is designated AMP-18.",
"AMP-18 genomic DNA, cDNA and the AMP-18 protein are sequenced for human, mouse and pig.",
"The AMP-18... | Patent_10473524 |
[
[
"Method and apparatus for enhanced particle collection efficiency",
"A method and apparatus for particle collection (30) that is characterized by co-aerosolizing fluids (62) into an air stream (34) containing the particles to be analyzed to significantly enhance their collection and identification efficie... | Patent_10473530 |
[
[
"Determination of risk factors for cataract by aldose reductase genotype",
"This invention relates to methods for determining the risk of developing cataract in mammals, and, more specifically, in mammals, including humans, with non-insulin dependent diabetes.",
"The methods are performed by determini... | Patent_10473537 |
[
[
"Use of herpesviruses, herpesvirus proteins and nucleic acids encoding the proteins to inhibit ccr5-tropic hiv-1 infection and replication",
"It has been discovered that herpesviruses can trigger an increase in the production of HIV-suppressive chemokines, and that these chemokines block the CCR5 receptor... | Patent_10473560 |
[
[
"Control of growth and repair of gastro-intestinal tissues by gastrokines and inhibitors",
"A novel group of gastrokines called Gastric Antrum Mucosal Protein is characterized.",
"A member of the group is designated AMP-18.",
"AMP-18 genomic DNA, cDNA and the AMP-18 protein are sequenced for human... | Patent_10473571 |
[
[
"Separate control line for maintenance of repeaters of a digital subscriber line",
"Digital subscriber line (DSL) system comprising a digital subscriber line access multiplexer (DSLAM, 20) having a line trunk unit (LTU, 13), a repeater container (23) containing a repeater (16) and a customer premises equi... | Patent_10473639 |
[
[
"Lipstick mechanism or the like with frictional braking",
"This lipstick mechanism comprising an inner tubular sleeve (1), an outer tubular sleeve (3), a stick-holder cup (2) arranged in the inner tubular sleeve (1).",
"The inner sleeve comprises a cylindrical base (12) in the wall (21) in which are f... | Patent_10473659 |
[
[
"Method of treatment",
"This invention relates to a method of maintaining glycemic control in diabetic hypertensive patients which comprises administering carvedilol to a subject in need thereof."
],
[
"1.A method of maintaining glycemic control in diabetic hypertensive patients which comprises ad... | Patent_10473667 |
[
[
"G protein-coupled receptor structural model and a method of designing ligand binding to g protein-coupled receptor by using the structural model",
"The present invention provides a method for constructing a structural model of a complex that a G protein-coupled protein receptor forms with a ligand capabl... | Patent_10473681 |
[
[
"Hardware design using evolution algorithms",
"The design of a hardware component such as a digital filter is optimized by taking an initial population of filter designs and encoding them as chromosomes.",
"The fitness of each chromosome is then evaluated and parent chromosomes are then selected based... | Patent_10473685 |
[
[
"Immunoglobulin construct containing tumor- specific p53bp2 sequenes for eliciting an anti-tumor response",
"The present invention provides immunoglobulins specific for p53BP2 ligand polypeptides.",
"In a preferred embodiment, the present invention provides a variant of an immunoglobulin variable doma... | Patent_10473691 |
[
[
"Methods and apparatus for treating diseased tissue",
"Methods and devices for treating diseased tissue using ultrasound waves are provided.",
"The ultrasound waves are not focused, and advantageously are administered in conjunction with radiation therapy and/or chemotherapy.",
"The methods and de... | Patent_10473712 |
[
[
"Carburized and quenched member and method for production thereof",
"A carburizing and hardening method enhances strength while sufficiently reducing hardening strain, without increasing the production cost, and a carburized and hardened member produced thereby.",
"The raw material is an alloy steel w... | Patent_10473716 |
[
[
"Oral absorbed drugs",
"Orally nonabsorbed or poorly absorbed drugs may be converted to orally absorbed prodrug derivatives by derivatization of free functional groups selected from amino, hydroxyl, mercapto, phosphate and/or carboxyl with groups sensitive to mild basic conditions such as 9-fluorenylmetho... | Patent_10473721 |
[
[
"Industrial robot",
"A communication system for safe wireless control of an manoeuvrable object (3), comprising a control unit (1) having means for operating the object and receiving means for wireless information, and a portable operating unit (2) having sending means for wireless information, where said... | Patent_10473733 |
[
[
"Isolated human ras-like proteins acid molecules encoding these human ras-like proteins, and uses thereof",
"The present invention provides amino acid sequences of polypeptides that are encoded by genes within the human genome, the Ras-like protein polypeptides of the present invention.",
"The present... | Patent_10473737 |
[
[
"Novel modulator of non-genomic activity of nuclear receptors (mnar) and uses thereof",
"A novel protein modulator of non-genomic activity of nuclear receptors (MNAR), which interacts with estrogen receptors alpha (ERα), estrogen receptor beta (ERβ), and other nuclear receptors, was discovered and charact... | Patent_10473753 |
[
[
"Mobile sleeve structure for maintaining spatial relationship between vertebrae",
"An arrangement and method of use for supporting a plurality of vertebrae in a cervical column in predetermined spatial relation and for facilitating a spinal fusion procedure.",
"A plurality of templates (12, 14) couple... | Patent_10473759 |
[
[
"Automatic optimization of a splice loss estimator for optical fiber splicers",
"A technique is provided for automatic optimization of a splice loss estimator of a fiber splicer (1), where the splice loss estimator is adapted, in a splice loss estimation procedure, to estimate the splice losses (Lti) of s... | Patent_10473766 |
[
[
"Alphanubeta3 integrin-binding polypeptide monobodies and their use",
"The present invention relates to a polypeptide monobody which includes a modified acid sequence and renders the polypeptide monobody able to bind selectively to αvβ3 integrin.",
"Fusion proteins and conjugates which include the pol... | Patent_10473770 |
[
[
"Controller of internal combustion engine",
"Disclosed is an internal combustion engine controller, which is used with a fuel-based torque-on-demand control type, multi-cylinder, direct-injection internal combustion engine to provide an air flow rate control means that excels in response and convergence."... | Patent_10473772 |
[
[
"Stabilised interbody fusion system for vertebrae",
"The invention concerns a stabilised interbody fusion system for vertebrae, of the type comprising an interbody implant (4) designed to be inserted in the intervertebral space defined between two neighbouring vertebrae to be mutually secured, so as to re... | Patent_10473773 |
[
[
"Actuator assembly",
"An actuator assembly comprises a first member (6a,6b), a second member (4a,4b) and at least one connecting arm (10a,10b) of fixed length connecting the first and second members (6a,6b;4a,4b).",
"The assembly also comprises an actuator (12,14) which is operably engaged with the fi... | Patent_10473798 |
[
[
"Installation for measuring pressure of at least one aeroplane wheel tyre",
"The invention concerns and installation (12) for measuring the pressure of at least one tyre of a wheel (16, 24) of an aeroplane (10) comprising: at least a pressure measuring sensor (26) associated with a tyre; a unit (30) opera... | Patent_10473808 |
[
[
"Arc welding method",
"A method for controlling an arc welding equipment (21) used in a welding operation and adjustable by varying at least one welding parameter value, comprising the steps of determining said at least one welding parameter while using a theoretical model (24) of the welding process asso... | Patent_10473819 |
[
[
"Trusted authorization device",
"A trusted display (18) of a trusted authorization device (TAD) (10) displays on a trusted display (18) first information about a transaction to be authorized by a user (14) using a trusted keypad (20).",
"The TAD (10) generates (208) a random number (R); generates (121... | Patent_10473842 |
[
[
"Filter element, method for manufacture thereof, and filter using said element",
"The invention provides a filter element exhibiting an excellent deodorizing and adsorbing function with low pressure drop, as well as to a method for its manufacture and a filter using the element, and this is achieved by a ... | Patent_10473866 |
[
[
"Telephone and method for converting sound to image and displaying on screen thereof",
"A telephone and a method for converting sound to image and displaying on a screen thereof are disclosed.",
"Receiver receives a call signal.",
"Storage section stores call sound information.",
"Call sound o... | Patent_10473872 |
[
[
"Process for producing foarmed articles and the foamed article",
"It is an object of the present invention to provide a method for manufacturing a high-quality foam that can be produced stably on an industrial scale and that affords excellent thermal insulation as a foam with thin cell membranes, which me... | Patent_10473881 |
[
[
"Protein analysis",
"A method of forming an array of proteins selected from antigens or antibodies; said method comprising the steps of (i) expressing in a recombinant cell, a fusion protein which comprises either (a) an antigen or (b) an antibody binding protein, fused to a peptide having up to 50 amino ... | Patent_10473882 |
[
[
"Medicament dispenser",
"Apparatus for dispensing medication to a patient, said apparatus comprising: (a) an electronic medicament dispenser (2) with one or more data-transfer elements; (b) a docking station (6) with one or more corresponding data-transfer elements; and (c) a wireless communications modul... | Patent_10473899 |
[
[
"Method for pressure-and flow-preventive fixing of pipes in a well",
"A method for pressure- and flow-prevetive fixing of well pipes, preferably liners and casings (14, 54, 60, 66) in a well when drilling the well, wherein the method also may be used in a completed well in order to place a pressure- and f... | Patent_10473907 |
[
[
"Tire components having improved modulus",
"A tire component including a tire tread that comprises a vulcanized rubber, a rubber filler, and a modified rubber, where the modified rubber i) contains pendent or functional groups that contain carboxylic acid or anhydride groups, ii) has pendently grafted the... | Patent_10473912 |
[
[
"Method and apparatus for fast w-cdma acquisition",
"Apparatus for classifying a ray is disclosed, wherein the ray is received at a code division multiple access receiver.",
"The apparatus comprises: means for receiving a neighbouring cell list comprising at least information concerning scrambling cod... | Patent_10473922 |
[
[
"Endosteal electrode",
"An implantable tissue-stimulating device comprising an elongate electrode carrier member (11) having a plurality of electrodes thereon.",
"The electrodes are preferably disposed in a linear array on the carrier member (11) and are adapted to apply a preselected tissue stimulati... | Patent_10473925 |
[
[
"Alumina agglomerates, the preparation method thereof and use of same as an absorbent or catalyst carrier",
"The invention relates to alumina agglomerates of the type obtained by dehydrating an aluminium oxyhydroxide or hydroxide, agglomerating the alumina thus obtained, hydrothermally treating the agglom... | Patent_10473938 |
[
[
"Adjustment Method, Particularly a Laser Adjustment Method, and an Actuator Suitable for the Same",
"The invention relates to an adjustment method, especially for the adjustment of optical or fibre optical components, whereby a partial region of an actuator is locally heated in such a defined manner that ... | Patent_10473940 |
[
[
"Adjustment Method, Particularly a Laser Adjustment Method, and an Actuator Suitable For the Same",
"The invention relates to an adjustment method, especially for adjusting optical or fibre optical components.",
"Said method involves locally heating, in a defined manner, at least one first partial reg... | Patent_10473941 |
[
[
"Bioluminescent plants and methods of making same",
"A genetically modified plant cell containing a heterologous nucleotide sequence encoding a bioluminescent polypeptide, which is expressed in amount sufficient to produce at least about 750,000 photons of visible light/mm2/second, is provided, as is a vi... | Patent_10473945 |
[
[
"Facilitating legal interception of ip connections",
"A method of facilitating the legal interception of IP connections, where two or more terminals can communicate with each other over the Internet using IPSec to provide security.",
"The method comprises allocating to each terminal T1,T2 a public/pri... | Patent_10473964 |
[
[
"Preparation containing polydimethylsiloxane for nail, cartilage,bone, joint,muscle and tendon disorders",
"The present invention relates to a topical preparation that contains polydimethylsiloxane for the external treatment of nail, cartilage, bone, joint, muscle and tendon disorders.",
"The preparat... | Patent_10473968 |
[
[
"Method for transferring information and associated network transition units",
"Disclosed is a method wherein operative, administrative and maintenance functions are defined initially for a time-division multiplex oriented communication network (12).",
"In order to utilize said functions in a packet-o... | Patent_10473969 |
[
[
"Switching contact arrangement for a power switch",
"The invention relates to a switching contact arrangement for a power switch, which is characterized in that a plurality of contact levers are pivoted on a contact support (30) by means of a bearing pin (37) and that the contact support (30) is provided ... | Patent_10473972 |
[
[
"Method for determining effective parathormone activity in a sample",
"Method for determining the active parathyroid hormone content in a sample, wherein the component of parathyroid hormone polypeptide chains which are oxidized at one or more sites near the PTH receptor binding structure having the amino... | Patent_10473975 |
[
[
"Immunotherapeutic combinations for the treatment of tumours that overexpress gangliosides",
"The present invention is related to the field of immunology and more specifically with immunotherapeutic combinations used for controlling growth and/or cell proliferation of tumors.",
"By means of the presen... | Patent_10473978 |
[
[
"Device and method for data flow exchange between a client device and a server",
"The invention concerns a device for data flow exchange between a client device (3) provided with an Internet file and a server (4) adapted to provide on request from the client device (3), at least a service for transforming... | Patent_10473984 |
[
[
"Method for eliminating metal halides that are present in a liquid or gaseous, organic or non-organic effluent",
"The invention relates to a method for eliminating metal halides which are present in a liquid or gaseous, organic or non-organic effluent.",
"According to the invention, the elimination is... | Patent_10473987 |
[
[
"Semi-continuous photochemical method and device therefor",
"In the photochemical synthetic process in semi-continuous mode according to the invention, a reactor comprising two zones is used, the radiating portion of the lamp(s) being totally immersed in a first zone which is completely filled with reacti... | Patent_10473993 |
[
[
"Modifying bitstreams",
"The bitstream includes digital codes representing an information signal.",
"At least one digital code is selected.",
"The code occupies a part of the bitstream which is to contain at least one watermark code which represents a watermark perceptible in the information signa... | Patent_10474013 |
[
[
"Dual core photonic crystal fibers(pcf) with special dispersion properties",
"An optical fibre for transmitting light, said optical fibre having an axial direction and a cross section perpendicular to said axial direction, said optical fibre comprising: (1) a first core region comprising a first core mate... | Patent_10474030 |
[
[
"Payment system",
"An electronic transaction payment system is provided having a vendor terminal associated with a vendor who provides goods or services to a purchaser, a vendor smart-card and a vendor smart-card reader for transmitting data to and receiving data from the vendor smart-card.",
"The sys... | Patent_10474050 |
[
[
"Filter cartridge for a filter assembly",
"A filter assembly including a replaceable filter cartridge for use with a reusable filter canister.",
"The filter cartridge includes a cylindrical filter media having top and bottom ends respectively mounted within top and bottom end caps.",
"The bottom e... | Patent_10474136 |
[
[
"Phage display vector, phages encoded thereby, and methods of use thereof",
"Filamentous bacteriophages are particularly efficient for the expression and display of combinatorial random peptides.",
"Two phage proteins are often employed for peptide display: the infectivity protein, pIII and the major ... | Patent_10474148 |
[
[
"Enantiomerically selective cyclopropanation",
"A method of forming a cyclopropane having enhanced chirality said method comprising reacting together: a symmetrical 1,2-dioxine of the formula (1), wherein X and Y are the same and are groups in which a carbon atom is bonded to the dioxine backbone; and a p... | Patent_10474149 |
[
[
"Pyrazolecarboxamides, intermediates therefor and pest control agent comprising the same as active ingredients",
"The present invention provides novel compounds with fungicidal and bactericidal activity as well as insecticidal and miticidal activity, which are useful for controlling hazardous biological o... | Patent_10474179 |
[
[
"Radio communication interface module, and its use in a device reading, processing, transmitting and operating on a bar code",
"A two-way radio communication interface module based on a predefined protocol for at least a user application, the module includes at least a radio circuit for transmitting and r... | Patent_10474185 |
[
[
"Method and divece for outputting data relating to information of a motor vehicle",
"In a method for outputting data relating to user selectable information regarding a motor vehicle, a plurality of user selectable options relating to the information are represented, and data relating to the selected opti... | Patent_10474189 |
[
[
"Device for the production of an extruded profile",
"A food product comprising a sugar wafer preferably cone-shaped having a filling of a substantially water-free fat-based confectionery material and a process for preparing it which comprises filling the sugar wafer with the substantially water-free fat-b... | Patent_10474197 |
[
[
"Trolley wheel assembly",
"A trolley includes a front wheel assemblie (12) and a rear wheel assembly both assemblies mounted on a trolley frame.",
"The front wheel assembly (12) includes a fixed wheel assembly (24) to aid control of the trolley.",
"In one form the front wheel assembly (12) include... | Patent_10474216 |
[
[
"Method for obtaining data relating to the sentiment on a stock exchange",
"The present invention relates to a method for obtaining data relating to the sentiment of transactions on an (electronic) stock exchange, comprising the following steps of:—entering by a potential principal for stock exchange tran... | Patent_10474262 |
[
[
"Specific detection of proteolytic enzymes",
"Specific detection of proteolytic enzymes is achieved by extinguishing dye fluorescence by the amino acid tryptophan.",
"Tryptophan is disposed on one side of the cutting site of a proteolytic enzyme while an amino acid marked with a dye is arranged on the... | Patent_10474263 |
[
[
"Modifiable assembly of microscopic apertures",
"The invention concerns a modifiable assembly of microscopic apertures comprising several plates (100, 110) coated with an opaque deposition except on transparent parts, for example (101, 114, 115), capable of sliding relative to one another, separated by an... | Patent_10474269 |
[
[
"Patent application cmos imager for cellular applications and methods of using such",
"Systems, methods and devices related to detecting and transmitting images.",
"Imaging systems and devices, as well as methods of using such that are provided herein include flicker detection and/or correction; and/o... | Patent_10474275 |
[
[
"Surface profile measurement",
"An imaging system scans a light source output over an object to be imaged, and receives the reflected signal in a multi-region light detector, wherein different regions of the object to be imaged are imaged onto different regions of the detector by the receiving optics, and... | Patent_10474293 |
[
[
"Supported double layer structure for displaying a nucleic acid associated with a protein",
"The invention concerns novel biosensors, in particular a support for displaying nucleic acids and for detecting both the presence of nucleic acids in a sample and the linkage between proteins and nucleic acids, as... | Patent_10474299 |
[
[
"Server-based browser system",
"A server-based browser system provides a server-based browser and a client system browser.",
"The client browser reports the position of a user click on its display screen, hotspot ID, or URL to the server-side browser which retrieves a Document Object Model (DOM) model... | Patent_10474300 |
[
[
"Thiazolyl-substsituted carbocylic 1,3-diones as pesticidal agents",
"The present invention relates to novel hetaryl-substituted carbocyclic 1,3-diones of the formula (I) in which A, B, Q1, Q2, Q3, Q4, G, m and Het are as defined in the description, to a plurality of processes for their preparation and to... | Patent_10474308 |
[
[
"Reversible life raft comprising removable shelters",
"Rescue and safety device, permitting the reversibility of a rescue raft, shelter (s)-tent (s), consists of an inflated grille provided with unlockable, spreadable and lockable openings, handles and safety belts.",
"On this grille are fixed bags (1... | Patent_10474317 |
[
[
"System for finding vehicles parked inside buildings and roofed surfaces",
"To find an automobile or another vehicle that it was left parked, in spaces intended for such purpose, by means of widely used current technology, of simple use and without adding accessories to the vehicle.",
"Several devices... | Patent_10474366 |